Traditional Chinese medicine composition for treating sepsis as well as preparation method and application thereof

Abstract

The invention discloses a traditional Chinese medicine composition for treating sepsis as well as a preparation method and application of the traditional Chinese medicine composition. The traditional Chinese medicine composition is prepared from 1-15 parts of sulfur, 3-25 parts of calamitas urinae hominis, 1-20 parts of wine-treated rhubarb, 10-30 parts of fructus cannabis, 10-50 parts of radix bupleuri, 5-30 parts of scutellaria baicalensis, 5-25 parts of folium isatidis, 5-25 parts of peach kernels, 5-30 parts of salvia miltiorrhiza, 5-30 parts of angelica sinensis, 5-30 parts of sophora flower charcoal, 5-25 parts of fructus gardeniae praeparatus and 5-60 parts of wine-treated fructus corni. According to the invention, through a combined mode of'monarch drug targeted oral administration and synergistic decoction of minister, assistant and guide drugs', the whole-course intervention of'upstream inhibition, midstream blocking and downstream regulation 'on sepsis immune disorder can be realized, meanwhile, the functions of warming yang, dispelling cold, cooling blood and detoxifying are taken into account, and the pro-inflammatory / anti-inflammatory balance is successfully recovered to a physiological state; the action mode is superior to that of a traditional single anti-inflammatory strategy and a traditional Chinese medicine application mode of whole-prescription decoction. In a standard animal model, the traditional Chinese medicine composition definitely shows the dose-dependent extremely remarkable effects of anti-inflammation, immunoregulation, multi-organ protection and overall condition improvement, the curative effect of a high-dose group of the traditional Chinese medicine composition is equivalent to that of a classic potent medicine, and the traditional Chinese medicine composition has more advantages in the aspects of recovering immune homeostasis, warming yang and strengthening the body resistance.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine technology, and in particular to a traditional Chinese medicine composition for treating sepsis, its preparation method, and its application. Background Technology

[0002] Sepsis is caused by an imbalance in the host response induced by infection, leading to life-threatening organ dysfunction and is one of the leading causes of death among critically ill patients worldwide. Its pathological mechanism is complex, with the core being the dysregulation of the body's immune inflammatory response, manifested as the excessive release of pro-inflammatory mediators (such as TNF-α, IL-1β, and IL-6) and insufficient anti-inflammatory mechanisms (such as IL-10), triggering a "cytokine storm" that ultimately leads to vascular endothelial damage, microcirculatory disturbances, and progressive damage and even failure of multiple organs such as the heart, liver, lungs, and kidneys.

[0003] Currently, standardized treatment in modern medicine focuses on early anti-infection measures, adequate fluid resuscitation, vasoactive drug support, and organ function replacement. However, this strategy primarily emphasizes controlling the source of infection and maintaining macroscopic vital signs, lacking safe and effective specific interventions for already initiated and uncontrolled systemic inflammatory responses, immune dysregulation, and the resulting early organ parenchymal damage. Antibiotics cannot modulate the host's immune response, while potent anti-inflammatory drugs (such as high-dose glucocorticoids) carry risks such as inducing secondary infections and exacerbating metabolic disorders, highlighting the limitations of current treatments in the crucial aspect of "modulating the host response."

[0004] Traditional Chinese medicine (TCM), based on the theories of "holistic view" and "syndrome differentiation and treatment," demonstrates unique potential in regulating complex inflammation and immune responses through its synergistic effects involving multiple components, targets, and pathways. It is considered a valuable supplementary strategy in the comprehensive treatment of sepsis. In TCM, sepsis is often categorized under "febrile diseases," "blood disorders," and "collapse." The main clinical symptoms in the pre-shock phase include chills and high fever secondary to purulent infections; rapid breathing accompanied by tachycardia, severe respiratory distress, and a sense of impending death; hypotension; constipation; decreased urine output (oliguria); altered mental status; pale, cold, and clammy skin with mottled changes or petechiae and ecchymoses; and multiple organ dysfunction. Currently, clinical treatments commonly employ methods such as clearing heat and detoxifying, promoting blood circulation and removing blood stasis, purging the bowels, and tonifying qi and consolidating the body. However, existing publicly available TCM compound prescriptions or research often focus on single pathogenesis such as intense heat and toxicity or bowel obstruction, and the approach to medication often leans towards large doses of bitter and cold herbs to clear heat or to purge the bowels. While such treatments can alleviate some symptoms, they generally overlook a key pathogenesis: sepsis patients often have a constitutional basis of "deficiency of Yang Qi and susceptibility to cold and dampness" due to congenital deficiencies or acquired imbalances. It is precisely this internal "cold and damp" environment and weakened vital energy that makes it easy for external heat toxins to penetrate deeply and combine with internal dampness, leading to prolonged illness, difficulty in tissue repair (prone to ulceration), and rapid depletion of vital energy (progressing towards shock). Clinically, patients in the pre-shock stage exhibit contradictory signs of "alternating chills and high fever" and "high fever with cold and clammy extremities," which are external manifestations of this complex pathogenesis of "cold and dampness stagnating and transforming into heat, and deficiency of vital energy with lingering pathogenic factors."

[0005] Therefore, both existing TCM and Western medicine strategies have significant shortcomings in addressing the critical "window period" of the pre-septic shock phase: modern medicine lacks safe and effective host response modulators; while existing TCM solutions mostly focus on "clearing heat, detoxifying, and purging," making it difficult to achieve early "interception and reversal" of the complex pathogenesis of "cold-damp constitution - heat toxin manifestations - blood stasis and visceral congestion - qi and yin depletion."

[0006] In light of this, there is an urgent clinical need to develop a comprehensive treatment approach based on innovative understanding of TCM pathogenesis. This approach integrates the principles of "warming and transforming internal cold and dampness to treat the root cause, clearing heat and toxins to reduce inflammation to treat the symptoms, opening and dispersing the bowels to cool the blood and dissipate blood stasis to unblock the meridians, and astringing and calming the five internal organs to prevent collapse." This approach should complement modern conventional treatments, be specifically designed for the pre-shock stage of sepsis, and aim to regulate immune-inflammatory imbalances through multiple targets, protect vascular endothelium and multi-organ function, thereby preventing the progression of the disease to shock and multiple organ failure. This invention is designed to address this pressing clinical need. Summary of the Invention

[0007] Purpose of the Invention: This invention aims to provide a traditional Chinese medicine composition specifically for the treatment of sepsis and its application. This composition targets the core pathogenesis of sepsis and the pre-septic shock stage, characterized by "suppuration and toxicity, easy ulceration due to cold and dampness, and stagnation due to heat and toxins." Symptom-oriented and treatment-centered, it achieves multi-target intervention through precise combination of principal, assistant, and adjuvant herbs to prevent the disease from progressing to shock and multiple organ failure.

[0008] Specifically, the objectives of this invention include: (1) In response to the suppuration and infection of sepsis, the tendency of cold and dampness to ulcerate, and the stagnation of heat and toxins, a combination of traditional Chinese medicine is provided to warm and dispel cold and dampness, clear heat and detoxify, stop bleeding and remove blood stasis, and promote tissue regeneration. Sulfur warms the yang and dispels cold to consolidate the root cause, targeting the pathogenesis of deficiency and coldness that is prone to ulceration due to cold and dampness; human placenta clears heat and removes blood stasis to treat the symptoms, specifically targeting the symptoms of heat and toxins stagnation and suppuration and infection.

[0009] (2) For symptoms of sepsis such as "chills and high fever secondary to purulent infection", a combination of Chinese medicine that can harmonize the Shao Yang and regulate cold and heat is provided. Bupleurum promotes the flow of Qi to relieve chills, and Scutellaria clears internal heat to reduce high fever. The two work together to quickly relieve the symptoms of alternating chills and heat and stabilize the body's response.

[0010] (3) For symptoms of sepsis such as "rapid breathing accompanied by tachycardia, severe dyspnea and a feeling of impending death, hypotension, and constipation", a combination of traditional Chinese medicine that can promote bowel movement, consolidate the body, invigorate blood circulation, and nourish the heart is provided. Through ingredients such as Salvia miltiorrhiza, Angelica sinensis, peach kernel, hemp seed, wine-processed Cornus officinalis, and wine-processed rhubarb, the aim is to improve the body's condition of disordered qi and blood in the heart and lungs, combined with bowel obstruction, to quickly improve the heart pulse, stabilize circulation, and alleviate the feeling of impending death.

[0011] (4) In response to symptoms of sepsis such as "systemic inflammation and skin ecchymosis and petechiae", a combination of traditional Chinese medicine that can clear heat, cool blood, detoxify and eliminate ecchymosis is provided. By using Isatis tinctoria leaf, charred gardenia fruit, and charred Sophora japonica flower to cool blood and detoxify, and wine-processed rhubarb, Salvia miltiorrhiza, and peach kernel to invigorate blood and remove blood stasis, the aim is to control the systemic inflammatory response, eliminate ecchymosis and protect the function of vital organs.

[0012] (5) In response to symptoms such as "altered state of consciousness" in sepsis, a combination of herbal medicines that can clear the mind, open the orifices, remove blood stasis, and refresh the spirit is provided. By using Isatis tinctoria leaf and charred Gardenia jasminoides to clear the mind and purge fire, and wine-processed rhubarb and Salvia miltiorrhiza to promote bowel movement, remove blood stasis, clear the mind, and relieve irritability, the aim is to promote mental clarity and improve nerve function.

[0013] (6) A comprehensive solution for multiple organ dysfunction is provided, which is to "harmonize the five internal organs and strengthen the body's resistance". Angelica sinensis, rhubarb, hemp seed, and cornus officinalis all have the functions of nourishing the five internal organs, benefiting the five internal organs, and harmonizing the five internal organs. They can stabilize the internal environment, improve the body's tolerance and repair potential under severe infection stress, and achieve "eliminating pathogens without harming the body's resistance and strengthening the body's resistance to toxins".

[0014] The traditional Chinese medicine composition described in this invention, through the combined approach of "warming, clearing, unblocking, and tonifying," and addressing both the root cause and symptoms, provides a TCM treatment strategy with synergistic and complementary potential to conventional modern sepsis treatments (such as anti-infection, fluid resuscitation, and organ support). It aims to compensate for the limitations of conventional treatments, reduce the risk of disease progression, and improve patient prognosis by regulating immune inflammation, protecting vascular endothelium, and improving coagulation and microcirculation through multi-target regulation. It offers a novel treatment strategy for the early intervention and reversal of the pre-septic shock phase, based on the holistic concept of traditional Chinese medicine, considering both constitution and disease symptoms, and employing synergistic mechanisms.

[0015] Technical solution: To achieve the above objectives, the technical solution adopted by this invention is as follows: A traditional Chinese medicine composition for treating sepsis, comprising the following raw materials in parts by weight: Sulfur 1-15 parts, human placenta 3-25 parts, wine-processed rhubarb 1-20 parts, hemp seed 10-30 parts, bupleurum 10-50 parts, scutellaria baicalensis 5-30 parts, indigo leaf 5-25 parts, peach kernel 5-25 parts, salvia miltiorrhiza 5-30 parts, angelica sinensis 5-30 parts, charred sophora japonica flower 5-30 parts, charred gardenia 5-25 parts, wine-processed cornus officinalis 5-60 parts.

[0016] As a preferred embodiment, the above-described traditional Chinese medicine composition for treating sepsis comprises the following raw materials in parts by weight: Sulfur 1-10 parts, human placenta 5-15 parts, wine-processed rhubarb 1-10 parts, hemp seed 10-20 parts, bupleurum 10-30 parts, scutellaria baicalensis 5-15 parts, indigo leaf 10-20 parts, peach kernel 5-15 parts, salvia miltiorrhiza 10-20 parts, angelica sinensis 5-15 parts, charred sophora japonica flower 5-15 parts, charred gardenia fruit 5-15 parts, wine-processed cornus officinalis 10-30 parts.

[0017] As a preferred embodiment, the above-described traditional Chinese medicine composition for treating sepsis comprises the following raw materials in parts by weight: Sulfur 3-6 parts, human placenta 10-15 parts, wine-processed rhubarb 6-10 parts, hemp seed 15-20 parts, bupleurum 15-30 parts, scutellaria baicalensis 10-15 parts, indigo leaf 15-20 parts, peach kernel 10-15 parts, salvia miltiorrhiza 15-20 parts, angelica sinensis 10-15 parts, charred sophora japonica flower 10-15 parts, charred gardenia 10-15 parts, wine-processed cornus officinalis 15-30 parts.

[0018] As a preferred embodiment, the above-described traditional Chinese medicine composition for treating sepsis comprises the following raw materials in parts by weight: Sulfur 3 parts, human placenta 10 parts, wine-processed rhubarb 6 parts, hemp seed 15 parts, bupleurum 15 parts, scutellaria baicalensis 10 parts, indigo leaf 15 parts, peach kernel 10 parts, salvia miltiorrhiza 15 parts, angelica sinensis 10 parts, charred sophora japonica flower 10 parts, charred gardenia fruit 10 parts, and wine-processed cornus officinalis 15 parts.

[0019] Specifically, in the traditional Chinese medicine composition of the present invention: Sulfur is a natural element mineral of the chalcogen group, which is naturally occurring sulfur. After mining, it is heated and melted to remove impurities; or it is obtained by processing sulfur-containing minerals. Human urine sediment (or white matter) is a solid substance that naturally settles in the urine of healthy individuals. Wine-processed rhubarb is a processed product of rhubarb, which is the plant Rheum palmatum of the Polygonaceae family. Rheum palmatum L., Tangut rhubarb (chicken claw rhubarb) Rheum tanguticum Maxim.ex Balf. or medicinal rhubarb Rheum officinale Dried roots and rhizomes of Baill.; Hemp seeds are from the mulberry family plant, cannabis. Cannabis sativa The dried, ripe fruit of L.; Bupleurum is a plant of the Apiaceae family. Bupleurum chinense DC. or Narrow-leaved Bupleurum (Red Bupleurum) Bupleurum scorzonerifolium The dried root of Willd.; Scutellaria baicalensis is a plant belonging to the Lamiaceae family. Scutellaria baicalensis dried roots of Georgi; Isatis indigotica Fort., a plant belonging to the Brassicaceae family, is a type of woad plant. Peach kernels are the dried, mature seeds of the Rosaceae plants Prunus persica (L.) Batsch or Prunus davidiana (Carr.) Franch. Danshen refers to the dried root and rhizome of *Salvia miltiorrhiza* Bge., a plant belonging to the Lamiaceae family. It is harvested in spring and autumn, cleaned of dirt and sand, and then dried. Angelica sinensis is a plant belonging to the Apiaceae family. Angelica sinensis The dried root of (Oliv.) Diels; Sophora japonica charcoal is a processed product made from Sophora japonica flowers, which are the dried flowers and buds of the legume Sophora japonica L. Charred Gardenia is a processed product of Gardenia, which is the dried, ripe fruit of Gardenia jasminoides Ellis, a plant in the Rubiaceae family. Wine-processed Cornus officinalis is a processed product of Cornus officinalis, which is a plant of the Cornaceae family. Cornus officinalis The dried, ripe pulp of Sieb.et Zucc.

[0020] The pharmacological effects of each herb in the herbal composition of this invention are as follows: Sulfur: [Properties and Channels Entered] Sour, warm. Enters the Kidney and Large Intestine channels.

[0021] [Shennong's Classic of Materia Medica] It is mainly used to remove stagnant blood, treat malignant sores, hidden sores in the lower body, stop bleeding, and strengthen tendons and bones.

[0022]

Functions and Indications

[0023] [Modern Pharmacology] This product has a certain antitussive and anti-inflammatory effect on experimental bronchitis. It forms hydrogen sulfide in the intestine, stimulating the intestinal wall to increase peristalsis and thus providing a mild laxative effect.

[0024] Ren Zhongbai: [Properties and Channels Entered] Salty, cool. Enters the Lung, Heart, and Bladder channels.

[0025]

Indications

[0026] [Ben Cao Jing Shu] Ni Bai Yu is salty in taste, cool in nature, and non-toxic. It can purge excess fire in the liver, kidneys, triple burner, and bladder.

[0027] [Ben Cao Zheng] Burn and grind into powder, it is very effective in treating various damp ulcers, chancres and malignant sores, promoting tissue regeneration and relieving heat toxins.

[0028] Rhubarb in wine: [Properties and Channels Entered] Rhubarb is bitter and cold. It enters the spleen, stomach, large intestine, liver, and pericardium channels.

[0029] [Shennong's Classic of Materia Medica] It is used to treat blood stasis, amenorrhea, chills and fever, break up masses and accumulations, promote metabolism, regulate the middle burner and aid digestion, and harmonize the five internal organs.

[0030]

Functions and Indications

[0031] [Modern Pharmacology] Rhubarb has anti-infective effects and inhibits a variety of Gram-positive and Gram-negative bacteria. The most sensitive bacteria are Staphylococcus and Streptococcus, followed by Corynebacterium diphtheriae, Salmonella typhi and paratyphi, Streptococcus pneumoniae, Shigella dysenteriae, etc.

[0032] Hemp seeds: [Properties and Channels Entered] Pungent in flavor, neutral in nature. Enters the spleen, stomach, and large intestine channels.

[0033] [Shennong's Classic of Materia Medica] It treats five types of fatigue and seven types of injury, benefits the five internal organs, and removes cold qi from the blood.

[0034] [Collected Annotations on Materia Medica] It breaks up accumulations, stops numbness, and disperses pus.

[0035]

Functions and Indications

[0036] Bupleurum: [Properties and Channels Entered] Pungent, bitter, slightly cold. Enters the Liver, Gallbladder, and Lung channels.

[0037] [Shennong's Classic of Materia Medica] It mainly treats the heart and abdomen, removes stagnant qi in the intestines and stomach, eliminates food accumulation, removes cold and heat pathogens, and promotes metabolism.

[0038] [Modern Pharmacology] Bupleurum and its active ingredient, saikosaponin, have anti-inflammatory effects, which are related to promoting the function of the adrenal cortex system. Bupleurum also possesses sedative, tranquilizing, analgesic, antitussive, lipid-lowering, hepatoprotective, choleretic, intestinal smooth muscle stimulating, gastric acid secretion inhibiting, anti-ulcer, trypsin inhibitory, antimicrobial, uterine stimulating, metabolism-influencing, antitumor, antiepileptic, anti-radiation, and immune-boosting effects.

[0039] Scutellaria baicalensis: [Properties and Channels Entered] Bitter, cold. Enters the Lung, Gallbladder, Spleen, Large Intestine, and Small Intestine channels.

[0040] [Shennong's Classic of Materia Medica] Treats amenorrhea, malignant sores and carbuncles, and boils.

[0041]

Functions and Indications

[0042] [Modern Pharmacology] Modern animal experiments have shown that the decoction has anti-inflammatory, immune-boosting, sedative and antipyretic effects.

[0043] Indigo leaf: [Properties and Channels Entered] Bitter, cold. Enters the Heart and Stomach channels.

[0044] [Collected Annotations on Materia Medica] It is used to treat seasonal headaches, high fever, and mouth ulcers.

[0045]

Functions and Indications

Modern Pharmacology

[0046] Peach kernel: [Properties and Channels Entered] Bitter, sweet, neutral. Enters the Heart, Liver, and Large Intestine channels.

[0047] [Shennong's Classic of Materia Medica] It is used to treat blood stasis, amenorrhea, abdominal masses, and pathogenic factors.

[0048]

Functions and Indications

[0049] [Modern Pharmacology] Peach kernel decoction and extracts have analgesic, anti-inflammatory, antibacterial and anti-allergic effects.

[0050] Salvia miltiorrhiza: [Properties and Channels Entered] Bitter, slightly cold. Enters the Heart and Liver channels.

[0051] [Shennong's Classic of Materia Medica] It treats evil qi in the heart and abdomen, rumbling in the intestines like flowing water, accumulation of cold and heat, breaks up masses and removes lumps, and benefits qi.

[0052]

Functions and Indications

[0053] [Modern Pharmacology] Danshen (Salvia miltiorrhiza) can combat arrhythmia, dilate coronary arteries, increase coronary blood flow, regulate blood lipids, and resist atherosclerosis; it can improve microcirculation, enhance hypoxia tolerance, and protect the myocardium; it can dilate blood vessels and lower blood pressure; it can reduce blood viscosity, inhibit platelet aggregation, and counteract thrombosis; it can protect against hepatocyte damage, promote hepatocyte regeneration, and has an anti-liver fibrosis effect; it can improve renal function and protect against ischemic kidney injury. In addition, Danshen also has certain sedative, analgesic, anti-inflammatory, and anti-allergic effects.

[0054] Angelica sinensis: [Properties and Channels Entered] Sweet, pungent, warm. Enters the Liver, Heart, and Spleen channels.

[0055] [Collected Annotations on Materia Medica] It treats chills and fever, various malignant sores and wounds caused by metal. It warms the middle burner and relieves pain, removes obstructed blood, nourishes the five internal organs, and promotes muscle regeneration.

[0056]

Functions and Indications

[0057] [Modern Pharmacology] The aqueous extract significantly promotes the production of hemoglobin and erythrocytes in mice; Angelica sinensis and its sodium ferulate have significant antithrombotic effects; the extract of this product dilates isolated guinea pig coronary arteries, increases coronary blood flow, and has a significant protective effect against experimental myocardial ischemia. In addition, this product enhances the body's immunity, inhibits the proliferation of granulation tissue in the later stages of inflammation, resists lipid peroxidation, and has anti-tumor, antibacterial, and anti-radiation effects.

[0058] Sophora japonica flower charcoal: [Properties and Channels Entered] Sophora japonica flowers are bitter and slightly cold. They enter the Liver and Large Intestine channels.

[0059] [Explanation of Materia Medica] It is used to treat hemorrhoids, heart pain, red eyes, and kills intestinal parasites.

[0060]

Functions and Indications

[0061] [Modern Pharmacology] Antibacterial, coagulating, and hemostatic. Pharmacological studies have shown that the rutin and its aglycone quercetin contained within can maintain the normal tension of capillaries, reduce their permeability, and restore normal elasticity to capillaries that have bled due to increased fragility. Quercetin can dilate coronary vessels, improve myocardial circulation, enhance cardiac contractility and output, and reduce heart rate. It has anti-inflammatory, antispasmodic, and anti-ulcer effects, as well as inhibitory effects on bacteria, viruses, and fungi.

[0062] Gardenia jasminoides: [Properties and Channels Entered] Bitter, cold. Enters the Heart, Lung, and Triple Burner channels.

[0063] [Shennong's Classic of Materia Medica] It treats evil qi in the five internal organs, white leprosy, red leprosy, and sores.

[0064]

Functions and Indications

[0065] [Modern Pharmacology] Gardenia extract can significantly inhibit the cytopathic effects of influenza A virus, PIV1, RSV, HSV, HSV1, and HSV2 viruses in vitro. This product has hepatoprotective and choleretic effects, promoting bile secretion and bilirubin excretion, and lowering blood bilirubin levels. Its decoction can reduce pancreatic amylase, promote pancreatic secretion, enhance the disease resistance of pancreatic cells during pancreatitis, and significantly increase normal hepatic blood flow. In addition, it also has antipyretic, analgesic, antibacterial, anti-inflammatory, sedative-hypnotic, and antihypertensive effects.

[0066] Wine mountain cornus meat: [Properties and Channels Entered] Sour, astringent, slightly warm. Enters the Liver and Kidney channels.

[0067] [Collected Annotations on Materia Medica] It treats chills and fever, warms the middle burner and lowers qi, promotes sweating, strengthens yin, benefits essence, calms the five internal organs, and opens the nine orifices.

[0068]

Functions and Indications

[0069] [Modern Pharmacology] Cornus officinalis enhances non-specific immune function and inhibits ascites cancer cells in in vitro experiments. It also has anti-experimental liver damage effects. It can increase white blood cell count in cases of leukopenia caused by chemotherapy and radiotherapy. Furthermore, it has antioxidant properties and a weak stimulant effect on the parasympathetic nervous system. The tannins it contains have astringent properties. Cornus officinalis injection can strengthen the heart and raise blood pressure. It can also inhibit platelet aggregation and prevent thrombosis. In addition, Cornus officinalis has antibacterial, antiviral, hypoglycemic, and diuretic effects. Fang Yi's analysis:

[0070] In traditional Chinese medicine, the pre-septic shock phase falls under the categories of "febrile disease" and "blood syndrome." In modern medicine, it is defined as a state of circulatory failure caused by infection-induced host response dysregulation, characterized by sequential organ dysfunction (SOFA score ≥2) but not yet progressing to the point of requiring vasoactive drugs to maintain blood pressure and persistent hyperlactatemia. Typical clinical manifestations include: chills and high fever secondary to purulent infection; rapid breathing accompanied by tachycardia, severe respiratory distress, and a sense of impending death; hypotension; constipation; decreased urine output (oliguria); altered mental status; pale, cold, clammy skin with mottled changes or petechiae and ecchymoses; and multiple organ dysfunction.

[0071] This invention systematically studies relevant literature on sepsis and deeply analyzes its clinical symptoms. Combining classical Chinese medicine theory with clinical practice, it proposes the pathogenesis, treatment principles, methods, and prescriptions for the pre-shock stage of sepsis.

[0072] Current clinical treatment approaches for sepsis largely focus on overt fever symptoms such as high fever, delirium, and ecchymosis, often employing large doses of bitter and cold herbs to clear heat, promote bowel movements, invigorate blood circulation, and detoxify. While this approach corrects imbalances, it often overlooks the underlying constitutional basis of "deficiency of Yang Qi and internal accumulation of cold and dampness" beneath the surface of intense heat and toxicity, stemming from congenital deficiencies or acquired malnutrition. Individuals with this constitution lack the body's ability to support its vital energy, making them susceptible to deep-seated internal invasion of external toxins after infection. Furthermore, the combination of pathogenic factors and internal dampness leads to putrefaction of flesh and blood, resulting in ulcers that are difficult to heal and oozing pus—this is the core pathogenesis of "prone ulceration." Clinical observations have revealed that the pre-shock stage of septicemia presents with signs such as "alternating chills and high fever" and "burning body with cold and clammy extremities," which are external manifestations of the complex pathogenesis of "cold and dampness stagnation transforming into heat, and intense struggle between the body's resistance and pathogenic factors." Therefore, the core pathogenesis of the pre-shock stage of septicemia can be summarized as "suppuration and infection, easy ulceration due to cold and dampness, and stagnation of heat and toxicity." This treatment plan is based on the principle of warming and dispelling internal cold and dampness to treat the root cause and expel toxins, and clearing away existing heat toxins to treat the symptoms and heal sores. It also uses strong remedies to open up the bowels, expel turbidity and remove blood stasis to guide the pathogenic toxins out and promote metabolism. Furthermore, it is supplemented with sour astringent and harmonizing ingredients to strengthen the body's foundation and prevent collapse. King medicine: Sulfur, Renzhongbai:

[0073] This formula uses sulfur and human placenta as the principal ingredients. One is warm and the other is cool, one is tonifying and the other is clearing. They complement each other and precisely target the core pathogenesis of "suppuration and infection, cold and dampness leading to easy ulceration, and heat and toxin stagnation". It takes into account both the root cause and the symptoms, and hits the disease site, laying the foundation for the whole formula to be effective.

[0074] Sulfur is sour and warm in nature, entering the kidney and large intestine meridians. Its potent properties are primarily focused on tonifying fire and assisting yang, dispelling cold and stagnation, playing a leading role in addressing the pathogenesis of "cold-dampness easily causing ulceration." The *Shennong Bencao Jing* states that it "mainly removes stagnant blood, treats malignant sores, treats hidden sores on the lower body, stops bleeding, and strengthens tendons and bones," highlighting its ability to remove blood stasis and stagnation, as well as to astringe sores, stop bleeding, and strengthen tendons and bones, thus preventing the prolonged skin ulceration caused by cold-dampness invasion. From a pathological perspective, cold-dampness stagnates on the skin surface; insufficient yang qi weakens its ability to warm and transform, leading to poor blood circulation, malnourishment of the skin, and easy ulceration, which is difficult to heal after ulceration. Sulfur warms and tonifies kidney yang, supporting the fundamental yang qi of the body. Abundant yang qi can warm and dispel cold-dampness, promote blood circulation, and curb the stagnation of cold-dampness at its source. Its sour and warm nature also has astringent and consolidating effects, stopping bleeding and astringing sores that have already ulcerated, preventing excessive pus discharge and further depletion of qi and blood. Modern pharmacological studies have shown that hydrogen sulfide formed by sulfur in the intestines can regulate the flow of qi and indirectly help the distribution of qi and blood. This corresponds to the traditional effects of "removing stagnant blood and strengthening tendons and bones," enhancing the ability to dispel cold and unblock stagnation, and taking into account both overall qi and blood harmony and local wound repair.

[0075] Human placenta (Ren Urinary Urinary Tract) is salty and cool in nature, entering the lung, heart, and bladder meridians. Its main functions are clearing heat and reducing fire, stopping bleeding and removing blood stasis, specifically targeting the pathogenesis of "suppuration and infection, and heat toxicity obstruction." The *Ben Cao Jing Shu* states that it "can purge excess fire from the liver, kidneys, triple burner, and bladder," and the *Ben Cao Zheng* also says that "when burned and ground into powder, it greatly treats various damp ulcers, chancres, and malignant sores, promotes tissue regeneration, and effectively resolves heat toxicity," clearly indicating its core functions of clearing heat and detoxifying, removing blood stasis and astringing sores in response to various damp ulcers and malignant sores. Internal heat toxicity is key to suppuration and infection; stagnation of heat toxicity leads to blood stasis, resulting in putrefaction, pus formation, and skin ulceration, accompanied by bleeding, swelling, and pain. Human placenta, being salty and cool, can directly clear excess heat toxicity from the triple burner and internal organs, thus eliminating the source of suppuration; its hemostatic and blood stasis-removing effects can break the stagnation of heat toxicity, promote the dissipation of blood stasis, and prevent the accumulation of stasis and toxins from worsening the sore surface. For various damp ulcers caused by excessive heat and toxins, human placenta can not only clear the surface heat and toxins, but also clear the internal fire in the internal organs, treating both the symptoms and the root cause. At the same time, its effect of promoting tissue regeneration can accelerate the healing of ulcers, complementing the effect of sulfur on cold and dampness.

[0076] The combination of these two herbs, sulfur, warms the yang and dispels cold to strengthen the body's foundation, addressing the pathogenesis of deficiency-cold that is prone to ulceration due to cold and dampness; human placenta clears heat and removes blood stasis to treat the symptoms, specifically targeting the excess heat syndrome of heat-toxin stagnation, suppuration, and infection. The combined use of warming and cooling herbs harmonizes the cold and heat, avoiding the drawbacks of solely warming yang and promoting heat, or solely clearing heat and damaging yang; the combined approach of tonifying and clearing balances the body's resistance to pathogens, both warming and tonifying kidney yang to enhance the body's ability to resist pathogens, and clearing heat and removing blood stasis to directly eliminate pathogens and repair ulcers. Both herbs closely adhere to the key pathogenesis of "suppuration and infection, easy ulceration due to cold and dampness, and heat-toxin stagnation," working together to warm and dispel cold and dampness, clear heat and detoxify, stop bleeding and remove blood stasis, and promote wound healing and tissue regeneration—the core essence of the principal herb.

[0077] The eleven herbs contained in this product are divided into three groups as assistant herbs, which are respectively targeted at the multi-system symptoms caused by secondary infection in the early stage of septic shock. They precisely match the core pathogenesis behind the symptoms, each performing its own function and synergistically enhancing the effect, assisting the principal herbs to further strengthen the effects of eliminating pathogens, supporting the body's resistance, and harmonizing the internal organs, effectively curbing the progression of the disease to the shock stage.

[0078] (1) For chills and high fever secondary to purulent infection: Bupleurum and Scutellaria baicalensis For chills and high fever secondary to purulent infection, Bupleurum and Scutellaria baicalensis, one clearing and the other soothing, harmonize the Shao Yang, which not only assists the principal drug in strengthening the heat-clearing and detoxifying power, but also unblocks the Qi mechanism to resolve the conflict between cold and heat, making them a key drug pair for relieving this core symptom.

[0079] Scutellaria baicalensis is bitter and cold in nature, and enters the lung, gallbladder, spleen, large intestine, and small intestine meridians. Its core functions are clearing heat and drying dampness, purging fire and detoxifying, making it a key herb for directly relieving heat toxicity and suppressing high fever. The *Shennong Bencao Jing* records its use for "bleeding amenorrhea, malignant sores and carbuncles, and ulcers," clearly demonstrating its therapeutic effect on heat toxicity in sores and ulcers, which aligns perfectly with the pathogenesis of suppurative infections: "internal accumulation of heat toxicity, necrotic tissue forming pus." The high fever in the pre-septic shock phase originates from intense heat toxicity permeating the three jiaos (upper, middle, and lower burners). Scutellaria baicalensis, being bitter and cold, can directly counteract excess heat toxicity in the internal organs and meridians, fundamentally blocking the generation and spread of heat toxicity and suppressing the escalation of the inflammatory storm. Its drying effect can clear dampness that is bound to heat toxicity in the body, preventing dampness lingering and worsening the infection. Combined with the salty and cool detoxifying power of the principal herb, human placenta, it strengthens the function of "clearing and resolving superficial heat." Modern pharmacology has confirmed that the decoction of Scutellaria baicalensis has significant anti-inflammatory, sedative and antipyretic effects. It can inhibit the reproduction of pyogenic bacteria and reduce systemic inflammatory response, providing clear pharmacological support for the relief of high fever symptoms.

[0080] Bupleurum is pungent, bitter, and slightly cold in nature, and it pertains to the Liver, Gallbladder, and Lung meridians. It is good at dispersing and regulating qi mechanism, reconciling the Shaoyang meridian, and promoting the renewal of qi and blood. It is specifically designed to break the pattern of shivering and the struggle between cold and heat. "Shennong Ben Cao Jing" states that it "treats the heart and abdomen, eliminates the stagnant qi in the stomach and intestines, food accumulations, and pathogenic factors of cold and heat", which fits the key pathogenesis of the early stage of septic shock, that is, "qi stagnation, the struggle between healthy qi and pathogenic factors cannot be vented", the symptom of shivering. It is not caused by simple cold pathogen, but actually due to the internal accumulation of cold-dampness and the stagnation of heat-toxin, resulting in the blockage of qi mechanism and the struggle between healthy qi and pathogenic factors in the Shaoyang meridian, unable to penetrate and be excreted. Bupleurum is pungent and dispersing, able to dredge the qi mechanism of the triple jiao, break the stagnant state of the mutual binding of heat-toxin and cold-dampness, provide a way for the exudation of pathogenic heat, make the struggle between healthy qi and pathogenic factors tend to be peaceful, and thus relieve shivering. Its slightly cold nature can not only assist Scutellaria baicalensis in clearing heat-toxin, but also avoid the stagnation of qi mechanism caused by single bitter-cold drugs, taking into account "clearing heat without stagnating pathogenic factors". At the same time, Bupleurum can harmonize the qi mechanism of the Liver and Gallbladder. The Liver governs dredging and dispersion. When the qi mechanism is smooth, qi and blood will circulate orderly, which can assist the monarch drug sulfur in warming and dredging qi and blood, promoting the dissipation of cold-dampness, and fundamentally reducing the pathological basis of the alternation of cold and heat. Modern pharmacology shows that Bupleurum and its active ingredient saikosaponin have anti-inflammatory and sedative effects, can synergistically regulate the body's inflammatory response with Scutellaria baicalensis, reduce the systemic metabolic disorder caused by high fever, and relieve the body's depletion caused by the struggle between cold and heat.

[0081] When the two are combined, Scutellaria baicalensis focuses on "clearing", directly clearing the excess of heat-toxin and cutting off the root cause of high fever; Bupleurum focuses on "dispersion", smoothing the stagnant qi mechanism and resolving the crux of shivering. One disperses and one clears, one ascends and one descends, jointly achieving the effects of reconciling the Shaoyang meridian, clearing heat and purging fire, and dispersing and regulating qi mechanism. It not only targets the nature of heat-toxin in suppurative infection, but also takes into account the pathogenesis characteristics of the struggle between cold and heat and qi stagnation, and at the same time fits the overall treatment principle of "warming and transforming cold-dampness, clearing heat-toxin" of the whole formula. Collaborating with the monarch drugs sulfur, human urine sediment, and other ministerial drugs, it can quickly relieve the symptoms of shivering and high fever, and at the same time prevent the internal invasion of heat-toxin and the progression of the disease, fully reflecting the formula-making wisdom of "taking both the root cause and the symptoms into account, and eliminating pathogenic factors without harming healthy qi".

[0082] (2)For those with shortness of breath accompanied by tachycardia, severe dyspnea and a sense of impending death, hypotension, constipation, etc.: Salvia miltiorrhiza, Angelica sinensis, Prunus persica, Cannabis sativa seed, Cornus officinalis processed with wine, Rheum officinale processed with wine This formula carefully selects six herbs, namely Salvia miltiorrhiza, Angelica sinensis, Prunus persica, Cannabis sativa seed, Cornus officinalis processed with wine, and Rheum officinale processed with wine, to form a refined compatibility with the core essence of "purging the fu-organs to secure the depletion, promoting blood circulation to nourish the heart", directly targeting the core pathogenesis of "disorder of qi and blood in the heart and lungs with concurrent fu-organ excess".

[0083] For hypotension and the collapse of heart qi, Cornus officinalis processed with wine is specifically used to secure the depletion and restore the pulse, stabilizing the fundamental of life, which is the key to saving the reversal and securing the depletion. Its nature is sour and warm, and it pertains to the Liver and Kidney meridians. It is good at tonifying the Liver and Kidney and astringing to secure the depletion. In this formula, it is like an "anchor stone", strongly tonifying the vital qi and astringing the dissipated qi and yin among a large number of herbs for purging and promoting blood circulation. Modern pharmacology has confirmed that it can strengthen the heart and raise blood pressure, directly counteracting the hypotension and sense of impending death caused by the decline of heart qi and the collapse of yang qi, and it is one of the main herbs for maintaining the foundation of life.

[0084] For tachycardia and hypotension (due to blood stasis in the heart vessels), Danshen (which enters the heart and liver meridians) and Danggui (which enters the heart, liver, and spleen meridians) work synergistically to invigorate blood and nourish the heart, thus improving circulation. Danshen invigorates blood, removes blood stasis, clears the heart, and relieves irritability. It can dilate coronary arteries, improve myocardial microcirculation, and combat arrhythmia. Danggui nourishes and invigorates blood, increasing coronary blood flow and combating myocardial ischemia. One is slightly cold and the other is warm, working together to remove blood stasis, unblock the meridians, and nourish the heart blood. They synergistically improve the heart's pumping function and blood supply, correcting the pathological states of "blood stasis obstructing the meridians" and "insufficient heart blood" behind tachycardia and hypotension. Rhubarb (which enters the pericardium meridian) and peach kernel (which enters the heart and liver meridians) act as vanguards in "breaking up blood stasis and expelling pathogens." Rhubarb removes blood stasis and unblocks the meridians, while peach kernel invigorates blood and removes blood stasis. Together, they "renew and eliminate" the old, working together to clear away heat toxins and blood stasis in the blood vessels, clearing obstacles for Danshen and Danggui to nourish and invigorate blood, and further improving systemic and myocardial microcirculation disorders.

[0085] For severe dyspnea and constipation (due to lung congestion and bowel obstruction), a combination of four herbs—rhubarb (processed with wine), hemp seed, peach kernel, and angelica root—forms a multi-layered "bowel-clearing and laxative" approach, addressing the root cause by clearing the bowels and purging the lungs. Rhubarb (processed with wine) acts as the main purgative, forcefully clearing away stubborn heat and stagnation in the intestines; hemp seed moistens the intestines and promotes bowel movements, its gentle nature preventing excessive purging from damaging body fluids, while also addressing constipation due to "blood deficiency and fluid depletion"; peach kernel and angelica root both have the effect of moistening the intestines and promoting bowel movements, and peach kernel can also stop coughing and relieve asthma, while angelica root can "remove internal obstruction of pathogenic blood," thus ensuring that bowel clearing simultaneously promotes blood circulation and moistens dryness. This combination achieves a "root cause" effect through powerful bowel movement: once the bowels are cleared, the upward-flowing turbid qi descends, directly relieving severe dyspnea caused by lung congestion (the lungs and large intestine are internally and externally related); at the same time, the elimination of intestinal toxins and stagnation greatly reduces systemic inflammation and toxin load, indirectly alleviating the inhibition of cardiopulmonary function.

[0086] In summary, this combination of six herbs utilizes the wine-processed Cornus officinalis to consolidate the body's vital energy and prevent collapse; Salvia miltiorrhiza and Angelica sinensis to nourish the heart and restore the pulse; and wine-processed Rheum palmatum and Prunus persica to break up blood stasis and unblock the meridians. Furthermore, wine-processed Rheum palmatum, Cannabis sativa seed, Prunus persica, and Angelica sinensis work together to clear the bowels. These herbs do not act in isolation but rather construct a synergistic therapeutic network: clearing the bowels to lower lung qi, invigorating blood to unblock the heart meridians, and consolidating the body's vital energy to stabilize the foundation. Clearing the bowels directly targets the pathological hub—constipation—and directly relieves breathing difficulties; invigorating blood and nourishing the heart improves cardiac function and circulation, counteracting hypotension and tachycardia; and consolidating the body's vital energy prevents the dissipation of vital energy during the attack on pathogens. The entire formula combines attack and tonification, treating both the symptoms and the root cause, precisely addressing the critical syndrome of intertwined "toxins, heat, blood stasis, obstruction, and deficiency."

[0087] For systemic inflammation, ecchymosis, and petechiae: Isatis leaf, rhubarb (processed with wine), charred gardenia fruit, charred sophora flower, salvia miltiorrhiza, angelica sinensis, and peach kernel. In traditional Chinese medicine, "systemic inflammation" (primarily manifested by high fever and delirium) falls under the categories of "heat entering the blood" or "dual burning of qi and blood." The formation of "ecchymosis and petechiae" originates from "excessive heat forcing blood to flow erratically"—excessive heat in the blood forces blood to overflow from the vessels and stagnate under the skin. The core pathological mechanism lies in the intertwining and lingering nature of "heat toxin" and "blood stasis."

[0088] Targeting the core "heat toxin" behind systemic inflammation, the combination of four herbs—Isatis tinctoria leaf, charred Gardenia jasminoides fruit, charred Sophora japonica flower, and rhubarb (processed with wine)—specifically clears heat toxin from the blood level, directly suppressing the rampant fire evil. Isatis tinctoria leaf, bitter and cold, enters the heart and stomach meridians, and is a key herb for clearing heat and detoxifying, cooling the blood and eliminating rashes. Its potent and focused effects directly clear heat toxin from the qi and blood levels, and it is particularly effective in treating heat toxin-induced rashes (i.e., ecchymosis), making it a core herb for treating systemic inflammation and skin bleeding points. Modern pharmacological research has also confirmed that Isatis tinctoria leaf has multiple effects, including broad-spectrum antibacterial, antiviral, anti-endotoxin, antipyretic, and anti-inflammatory properties, highly consistent with the traditional Chinese medicine principle of "clearing and detoxifying." Charred Gardenia jasminoides fruit, bitter and cold, enters the heart, lung, and triple burner meridians, and excels at clearing heat and purging fire, cooling the blood and stopping bleeding. Its properties are clearing and unblocking, able to clear the fire spreading throughout the three jiaos (upper, middle, and lower burners), guiding pathogenic heat downwards and outwards, especially effective in clearing various types of bleeding caused by blood heat (such as hematemesis and epistaxis), alleviating the situation of "excessive heat forcing blood to flow recklessly" from the root. In addition, its liver-protecting and bile-promoting effects can also help reduce the damage to the liver caused by systemic inflammation. Sophora japonica flower charcoal has a bitter and astringent taste, is slightly cold in nature, and enters the liver and large intestine meridians. Its function is to cool the blood and stop bleeding, clear the liver and purge fire. After being processed by charring, its hemostatic power is particularly strong. Because it enters the liver meridian (the liver is responsible for storing blood), it can clear the heat in the blood of the liver meridian, and has a direct astringent and hemostatic effect on bleeding caused by reckless blood heat. Modern research shows that Sophora japonica flower charcoal can enhance capillary resistance and restore its normal elasticity, which is crucial for preventing and reducing the formation of ecchymosis. Rhubarb in this formula is used for its heat-clearing, fire-purging, blood-cooling, and detoxifying effects. It enters the spleen, stomach, large intestine, and pericardium meridians, and is particularly good at clearing heat and toxins in the upper jiao blood. When combined with the aforementioned three herbs, it can enhance the overall efficacy of clearing heat toxins from the blood; at the same time, its "renewal" properties can open up pathways for the elimination of heat toxins from the body and promote the expulsion of pathogens.

[0089] For ecchymosis, petechiae, and microcirculatory disorders (blood stasis), a combination of rhubarb (processed with wine), salvia miltiorrhiza, peach kernel, and angelica sinensis is used to clear the meridians and prevent "heat and blood stasis" by cooling the blood, promoting blood circulation, and dispersing blood stasis. Rhubarb (processed with wine) has a strong and penetrating effect in "expelling blood stasis and clearing the meridians," breaking up the blockage caused by the mutual binding of blood stasis and heat in the blood vessels; at the same time, its laxative effect also helps to release blood stasis and heat from the lower abdomen. Salvia miltiorrhiza is bitter and slightly cold in nature, and enters the heart and liver meridians. It is an important medicine for promoting blood circulation, removing blood stasis, cooling the blood, and eliminating carbuncles. It can effectively improve microcirculation, inhibit platelet aggregation, and prevent thrombosis, directly targeting microthrombosis and coagulation dysfunction commonly seen in sepsis. Peach kernel is bitter and sweet in taste and neutral in nature, and enters the heart, liver, and large intestine meridians. It can both promote blood circulation and remove blood stasis, and also has the effect of moistening the intestines and promoting bowel movements. It can assist rhubarb (processed with wine) and salvia miltiorrhiza in enhancing the effect of removing blood stasis, and its moist texture can prevent excessive consumption of body fluids by blood-activating medicines. Angelica sinensis has a sweet and pungent taste and is warm in nature. It enters the liver, heart, and spleen meridians and excels at nourishing and activating blood. Among a large number of blood-cooling and blood-activating herbs, Angelica sinensis's "blood-nourishing" function is particularly crucial. It can nourish blood and calm the meridians, removing blood stasis without harming the body's vital energy, and repairing damaged blood vessels. At the same time, its anti-thrombotic and hematopoietic effects also help restore normal coagulation and blood flow.

[0090] These seven herbs work by "clearing" heat (Isatis leaf, charred gardenia, charred sophora flower, and wine-processed rhubarb) and "dispersing" blood stasis (wine-processed rhubarb, salvia miltiorrhiza, peach kernel, and angelica sinensis). This combination of clearing and dispersing, addressing both the symptoms and the root cause, provides core pharmacodynamic support for controlling systemic inflammatory responses and correcting coagulation disorders (manifested as ecchymosis).

[0091] (4) For changes in consciousness: Rhubarb (processed with wine), Isatis leaf, Gardenia fruit (processed with charred herbs), Salvia miltiorrhiza In response to altered states of consciousness (such as agitation, delirium, drowsiness, or even coma) that occur during the course of sepsis, Traditional Chinese Medicine (TCM) often diagnoses this as "heat entering the pericardium" or "blood stasis obscuring the sensory orifices," with the core pathogenesis being the disturbance of the mind by heat toxins and blood stasis. The selected four herbs—rhubarb (processed with wine), indigo leaf, charred gardenia, and salvia miltiorrhiza—are designed to "clear the heart and open the orifices, cool the blood and remove blood stasis," directly addressing the key pathogenesis of delirium and coma.

[0092] Both Isatis leaf and charred gardenia clear heat and purge fire, directly eliminating heat toxins. Isatis leaf is bitter and cold, entering the heart and stomach meridians. It excels at clearing heat and detoxifying, cooling the blood and eliminating spots. Its nature is potent and powerful, directly clearing heat toxins deep into the blood, especially relieving the disturbance of the central nervous system caused by high fever. With heat toxins cleared, the mind is at peace. Modern pharmacology has confirmed that it has significant anti-endotoxin, antipyretic, and anti-inflammatory effects, which can reduce the toxic damage of endotoxins to brain function from the source. Charred gardenia is bitter and cold, entering the heart, lung, and triple burner meridians. It purges fire and relieves irritability, cools the blood and detoxifies. It is especially good at clearing excess fire in the heart meridian and triple burner. It has a direct effect of relieving irritability and calming the mind for restlessness, agitation, and even delirium caused by heat disturbing the heart. Its downward nature can guide heat out through urination, opening an external pathway for heat to escape.

[0093] Rhubarb (processed with wine) and Salvia miltiorrhiza (Salvia miltiorrhiza) promote bowel movement and remove blood stasis, providing an outlet for pathogenic factors. Rhubarb, through its purgative effect, first helps to expel the stagnant heat and toxins from the intestines through feces, achieving a "bottom-up" effect. Once the bowel function is unblocked, the upward-flowing heat and toxins descend, directly reducing their impact on the heart and brain. Secondly, it removes blood stasis and unblocks the meridians, entering the pericardium meridian, effectively clearing heat and toxins from the upper burner, breaking up blood stasis, clearing the accumulation of heat and stagnation in the pericardium and brain, and unblocking the channels of the mind. Modern research suggests that its purgative effect helps to clear intestinal toxins and reduce secondary damage to the central nervous system caused by systemic inflammation. Salvia miltiorrhiza, bitter and slightly cold, enters the heart and liver meridians, promoting blood circulation, removing blood stasis, clearing the heart, and relieving irritability. It serves as a key link between "blood circulation" and "calming the mind": on the one hand, its effects of improving microcirculation and preventing thrombosis can alleviate cerebral microcirculatory disorders, laying a solid material foundation for mental recovery; on the other hand, its function of "clearing the heart and relieving irritability" can directly stabilize the mind. When combined with rhubarb, one dispels blood stasis and the other promotes purgation, together dissolving the "blood stasis and heat" that obstructs the sensory orifices. Assistant herbs: Angelica sinensis, rhubarb (processed with wine), hemp seed, and cornus officinalis (processed with wine).

[0094] Angelica sinensis, rhubarb (processed with wine), hemp seed, and cornus officinalis (processed with wine) are used as adjuvant herbs. The main purpose is to provide a comprehensive solution for multiple organ dysfunction, which is to "harmonize the five internal organs and strengthen the body's resistance to collapse". All four herbs have the functions of nourishing, benefiting, and harmonizing the five internal organs. They can stabilize the internal environment, improve the body's tolerance and repair potential under severe infection stress, and achieve the goal of "eliminating pathogens without harming the body's resistance and strengthening the body's foundation to support the body's resistance to toxins".

[0095] The preparation method of the traditional Chinese medicine composition for treating sepsis according to the present invention includes the following steps: (1) Take the following ingredients by weight: rhubarb, hemp seed, bupleurum, scutellaria, indigo leaf, peach kernel, salvia miltiorrhiza, angelica sinensis, charred sophora japonica flower, charred gardenia and cornus officinalis. Place them in a decoction vessel, add an appropriate amount of water and soak for 40-60 minutes. Then heat and decoct for 1 hour. Filter to obtain the Chinese herbal decoction for later use. (2) Take sulfur and human placenta in the proportion of weight, grind them into powder, and mix them with the medicine solution in step (1) in proportion.

[0096] As a preferred embodiment, the preparation method of the above-mentioned traditional Chinese medicine composition for treating sepsis includes the following steps: (1) Take the following ingredients by weight: rhubarb, hemp seed, bupleurum, scutellaria, indigo leaf, peach kernel, salvia miltiorrhiza, angelica sinensis, charred sophora japonica flower, charred gardenia and cornus officinalis. Place them in a decoction vessel, soak them in 5 to 20 times the amount of water for 10 to 60 minutes, then heat and decoct them 1 to 3 times, each time for 0.5 to 3 hours. Filter, combine the decoction extracts, and store them in a refrigerator at 4 degrees Celsius for later use. (2) Take sulfur and human placenta in the proportion of weight, grind them into powder, and mix them with the medicine solution in step (1) in proportion.

[0097] As a preferred embodiment, the preparation method of the above-mentioned traditional Chinese medicine composition for treating sepsis includes the following steps: (1) Take the following ingredients by weight: rhubarb, hemp seed, bupleurum, scutellaria, indigo leaf, peach kernel, salvia miltiorrhiza, angelica sinensis, sophora japonica flower, charred gardenia and cornus officinalis. Place them in a decoction vessel, soak them in 5 to 20 times the amount of water for 40 minutes, then heat and decoct for 40 minutes. Filter the liquid and store it in a refrigerator at 4 degrees Celsius for later use. (2) Take sulfur and human placenta in the weight proportions, grind them into powder, pass them through an 80-mesh sieve, mix them with the liquid in step (1), add a pharmaceutically acceptable carrier, and prepare a traditional Chinese medicine preparation.

[0098] Preferably, the dosage form of traditional Chinese medicine preparations is tablets, pills, powders, granules, or capsules.

[0099] Beneficial effects: Compared with the prior art, the present invention has the following advantages: (1) Multi-target, bidirectional regulation of immune homeostasis, achieving synergistic effects of "clearing heat" and "strengthening the body's resistance". The traditional Chinese medicine composition provided by this invention employs a differentiated preparation process of "powdering the principal herb for oral administration and decocting the adjuvant herbs," which ensures the stable dissolution of the effective components of the adjuvant herbs while retaining the core efficacy of the principal herb. The two work synergistically to exhibit a systemic and potent regulation of key immune pathways in sepsis. "Upstream" source control: It can significantly inhibit the release of the core initiating factor TNF-α in a dose-dependent manner (the high-dose group was comparable to dexamethasone, P>0.05), blocking the inflammatory cascade response at its source. Among them, the principal ingredient, Ren Zhong Bai, is salty and cool, clearing heat, cooling blood and detoxifying. When powdered and taken with water, the effective components can directly enter the blood and quickly exert their anti-toxic effects. Sulfur warms the yang and dispels cold. Its powdered form is more easily absorbed by the decoction, avoiding the dissipation of warming components during decoction. Together with the assistant ingredients, Da Qing Ye and Huang Qin, they strengthen the source inhibition of heat toxicity.

[0100] The "midstream" component possesses strong anti-inflammatory properties: it can significantly inhibit the excessive production of key pro-inflammatory factors IL-1β and IL-6 (P<0.01 compared with the model group in the medium and high dose groups), effectively curbing the amplification and spread of inflammatory storms. The detoxifying power of the principal ingredient, human placenta extract, complements the anti-inflammatory components of the assistant ingredients, while sulfur promotes blood circulation by warming the yang and unblocking the meridians, thus aiding in the metabolism and clearance of inflammatory factors.

[0101] Downstream immune support: It can significantly increase the level of the anti-inflammatory factor IL-10 (the high-dose group recovered to levels close to those of the sham-operated group, P>0.05), enhancing the body's own negative immune regulation ability. Sulfur tonifies fire and strengthens yang, while human placenta clears heat without harming the body's vital energy. The powdered and dissolved form of both avoids the destruction of their active ingredients by decoction. Together with the assistant herbs such as Cornus officinalis and Angelica sinensis, it achieves the immune regulation goal of "strengthening the body's resistance to eliminate pathogens".

[0102] Through synergistic effects, the traditional Chinese medicine composition (high-dose group) provided by this invention can restore the IL-6 / IL-10 ratio, a key indicator reflecting systemic inflammatory imbalance, to the normal range (P>0.05) with no statistical difference from the sham surgery group, achieving a qualitative change from "unidirectional inhibition of inflammation" to "bidirectional regulation and restoration of immune homeostasis". Its effect is better than that of dexamethasone alone.

[0103] This differentiated preparation process precisely matches the characteristics of the principal drug—sulfur is warm and dry, and prolonged decoction can easily lead to the volatilization of its components and its strong medicinal properties; human placenta is tough and difficult to fully dissolve its effective components when decocted. Powdering and taking it with water can maximize the retention of the efficacy of both drugs, while ensuring the synergistic effect of the effective components of the assistant and adjuvant drugs in the compound with the principal drug. This enables multi-level intervention on the pathogenesis of sepsis, namely "internal invasion of toxic heat, stagnation of cold and dampness, and obstruction of qi and blood," perfectly embodying the essence of traditional Chinese medicine treatment principle of "eliminating pathogens without harming the body's resistance, and supporting the body's resistance to eliminate pathogens."

[0104] (2) Protection of multiple organ functions and improvement of overall condition, reflecting the advantages of holistic treatment The process combination of "powdered principal herb taken orally + decocted assistant herbs" provided by this invention enables its immune regulation and warming and detoxifying effects to work synergistically and directly translate into a protective effect on the body as a whole and on vital organs. Significantly improved general condition and prognosis: During the acute phase, it significantly reduced weight loss in model animals in a dose-dependent manner (the effect of the high-dose group was comparable to that of dexamethasone), and during the recovery phase, it promoted weight regain (the high-dose group achieved positive weight gain within 7 days), indicating that it can effectively improve the metabolic consumption caused by sepsis and promote recovery. This effect is attributed to the rapid warming and strengthening effect of sulfur powder after oral administration, which can alleviate the symptoms of cold and clammy extremities and depletion of vital energy in the early stage of sepsis, forming a closed loop with the anti-inflammatory and organ-protecting effects of the adjuvant drugs.

[0105] The basis for providing multi-organ protection: The aforementioned improvement in liver (ALT, AST) and kidney (BUN, CRE) function indicators, together with the systemic anti-inflammatory, immunomodulatory, and detoxifying and protective effects of the principal drug demonstrated in this invention, constitute a complete causal logic chain—human placenta powder taken orally can directly remove endotoxins and reduce toxic damage to the liver and kidneys; sulfur warms and tonifies kidney yang, enhances the qi transformation function of the liver and kidneys, and together with the adjuvant drugs to control systemic inflammation, they jointly reduce damage to vascular endothelium and organs such as the liver, kidneys, and lungs.

[0106] The standardized "soaking-decoction-infusion" combination process not only ensures the stability and reliability of the efficacy of the adjuvant and assistant herbs, but also optimizes the absorption efficiency of the medicine by pulverizing the principal herb and infusing it with water. This allows Compound B to not only regulate immunity at the molecular level, but also improve the overall quality of life of animals and protect the function of multiple organs through the synergistic effects of warming yang, detoxifying, and anti-inflammatory. This meets the requirements of modern medicine for comprehensive treatment of sepsis and is in line with the concept of "holistic treatment" in traditional Chinese medicine.

[0107] (3) Clear dose-dependent effect and superior formulation efficacy The dose-response relationship is clear: In all observed indicators (inhibition of pro-inflammatory factors, enhancement of anti-inflammatory factors, restoration of immune ratio, and weight protection), the compound B of this invention showed a clear dose-dependent improvement trend. The high-dose group (30g / kg, containing the corresponding dose of the principal drug powder) achieved effects comparable to or even better than the positive control drug dexamethasone (2mg / kg) in all key indicators (in terms of restoring IL-10 and immune homeostasis), providing precise experimental evidence for clinical dosage selection. The method of powdering and dissolving the principal drug ensures precise and controllable dosage, avoiding fluctuations in the dose-response relationship caused by component loss during decoction, further improving the reliability of experimental data.

[0108] The formulation exhibits significant advantages: at the same dosage, Compound B of this invention is significantly superior to Compound A in inhibiting pro-inflammatory factors (TNF-α, IL-1β, IL-6), increasing anti-inflammatory factors (IL-10), correcting immune imbalance (IL-6 / IL-10 ratio), and improving body weight (P<0.05). This advantage stems not only from the optimized combination of medicinal ingredients and dosage ratios but also from the innovative process of "powdering the principal herb and dissolving it in water"—which preserves the targeted effect of the principal herb on the core pathogenesis while synergistically enhancing the effects with the decoction components of the adjuvant herbs. This avoids the problem of weakened efficacy of the principal herb caused by traditional whole-formula decoction, directly demonstrating the inventiveness of this invention.

[0109] The clear dose-effect relationship, good safety (reflected in animal weight recovery), and standardized "decoction + oral administration" combination process lay a solid pharmacodynamic foundation for the subsequent development of Compound B of this invention into a modern Chinese medicine preparation for the treatment of sepsis (such as granules, oral liquids, and matching principal drug powder oral administration), which is more in line with the needs of individualized clinical medication.

[0110] Therefore, this invention not only closely adheres to the treatment principles of "clearing heat and detoxifying, strengthening the body's resistance, and warming yang and unblocking stagnation" in traditional Chinese medicine theory, and precisely matches the medicinal properties of the principal herb by grinding it into powder and taking it by hand, but also provides multi-level and multi-indicator conclusive evidence in modern pharmacology, proving its excellent technical effects and clinical application potential under the guarantee of differentiated preparation process. Attached Figure Description

[0111] Figure 1 A comparative chart showing the results of serum alanine aminotransferase (ALT) levels in CLP-induced sepsis mice in each treatment group to restore liver function.

[0112] Figure 2 A comparative chart showing the results of serum AST levels in mice with CLP-induced sepsis in each treatment group to restore liver function.

[0113] Figure 3 A comparative chart showing the results of measuring serum urea nitrogen (BUN) levels in CLP-induced sepsis mice to restore renal function in each treatment group.

[0114] Figure 4 A comparative graph showing the results of serum creatinine level restoration in CLP-induced sepsis mice from different treatment groups.

[0115] Figure 5 A comparative chart showing the results of serum IL-1β level restoration in CLP-induced sepsis mice from different treatment groups.

[0116] Figure 6 A comparative chart showing the results of serum IL-6 level restoration in CLP-induced sepsis mice from different treatment groups.

[0117] Figure 7 A comparative chart showing the results of serum IL-10 measurements in mice recovering from CLP-induced sepsis in each treatment group.

[0118] Figure 8 A comparative chart showing the results of serum TNF-α measurement in mice recovering from CLP-induced sepsis in each treatment group.

[0119] Figure 9 A comparative graph showing the effects of different treatment groups on reducing cardiac pathological damage in CLP-induced sepsis mice.

[0120] Figure 10 A comparative diagram showing the effects of different treatment groups on reducing liver pathological damage in CLP-induced sepsis mice.

[0121] Figure 11 A comparative chart showing the effects of different treatment groups on reducing spleen pathological damage in CLP-induced sepsis mice.

[0122] Figure 12 A comparative chart showing the effects of different treatment groups on reducing lung pathological damage in CLP-induced sepsis mice.

[0123] Figure 13 A comparative chart showing the effects of different treatment groups on reducing kidney pathological damage in CLP-induced sepsis mice.

[0124] Figure 14 A comparative diagram showing the effects of different treatment groups on reducing cecal pathological damage in CLP-induced sepsis mice.

[0125] Figure 15 A comparative graph showing the effects of different treatment groups on changes in body weight in CLP-induced sepsis mice. Detailed Implementation

[0126] The present invention is further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. After reading this invention, any modifications of the invention in various equivalent forms by those skilled in the art fall within the scope defined by the appended claims. All raw materials used in this invention are commercially available. Example 1

[0127] 1. A traditional Chinese medicine composition (compound traditional Chinese medicine A), the raw material composition of which is: 10g peach kernel, 6g rhubarb (processed with wine), 3g sulfur, 15g cistanche, 15g lily bulb, 10g honeycomb, 15g five-finger peach, 15g pig manure bean, 15g hemp seed.

[0128] 2. A method for preparing a traditional Chinese medicine composition, comprising the following steps: (1) Place 10g of peach kernel, 6g of rhubarb, 15g of cistanche, 15g of lily, 10g of bee honeycomb, 15g of five-finger peach, 15g of pig manure bean, and 15g of hemp seed in a decoction pot, add 6 times the amount of water and soak for 60 minutes; then decoct in an automatic Chinese medicine decoction pot for 1 hour at a temperature above 100℃ throughout the process. The crude extract of Chinese medicine obtained is filtered to obtain the Chinese medicine decoction liquid, concentrated, and stored in a refrigerator at 4 degrees Celsius for later use.

[0129] (2) Grind 3g of sulfur into powder and mix it with the concentrated liquid from step (1). Example 2

[0130] 1. A traditional Chinese medicine composition for treating sepsis (compound traditional Chinese medicine B), the raw materials of which are: Sulfur 3g, human placenta 10g, wine-processed rhubarb 6g, hemp seed 15g, bupleurum 15g, scutellaria baicalensis 10g, indigo leaf 15g, peach kernel 10g, salvia miltiorrhiza 15g, angelica sinensis 10g, charred sophora japonica flower 10g, charred gardenia fruit 10g, wine-processed cornus officinalis 15g.

[0131] 2. A method for preparing a traditional Chinese medicine composition for treating sepsis, comprising the following steps: (1) Place 6g of rhubarb, 15g of hemp seed, 15g of bupleurum, 10g of scutellaria, 15g of indigo leaf, 10g of peach kernel, 15g of salvia miltiorrhiza, 10g of angelica sinensis, 10g of charred sophora japonica flower, 10g of charred gardenia, and 15g of cornus officinalis in a decoction vessel, add 6 times the amount of water and soak for 60 minutes; then decoct in an automatic Chinese medicine decoction vessel for 1 hour at a temperature above 100℃ throughout the process. The crude extract of Chinese medicine is obtained, filtered to obtain the Chinese medicine decoction liquid, concentrated, and stored in a refrigerator at 4 degrees Celsius for later use.

[0132] (2) Grind 3g of sulfur and 10g of human placenta into powder and mix with the concentrated medicine solution in step (1). Example 3

[0133] 1. A traditional Chinese medicine composition for treating sepsis, comprising the following raw materials: Sulfur 10g, human placenta 15g, wine-processed rhubarb 10g, hemp seed 20g, bupleurum 30g, scutellaria baicalensis 15g, indigo leaf 20g, peach kernel 15g, salvia miltiorrhiza 20g, angelica sinensis 15g, charred sophora japonica flower 15g, charred gardenia fruit 15g, wine-processed cornus officinalis 30g.

[0134] 2. A method for preparing a traditional Chinese medicine composition for treating sepsis, comprising the following steps: (1) Place 10g of rhubarb, 20g of hemp seed, 30g of bupleurum, 15g of scutellaria, 20g of indigo leaf, 15g of peach kernel, 20g of salvia miltiorrhiza, 15g of angelica sinensis, 15g of charred sophora japonica flower, 15g of charred gardenia, and 30g of cornus officinalis in a decoction vessel, add 8 times the amount of water and soak for 60 minutes; then decoct in an automatic Chinese medicine decoction vessel for 1 hour at a temperature above 100℃ throughout the process, remove the dregs, add 6 times the amount of water, decoct for 1 hour, filter, combine the filtrates, concentrate to obtain concentrated liquid, and store in a refrigerator at 4 degrees Celsius for later use.

[0135] (2) Grind 10g of sulfur and 15g of human placenta into powder, mix with the concentrated liquid in step (1), and prepare tablets, granules, pills or capsules with pharmaceutical carriers such as cyclodextrin and starch. Example 4

[0136] 1. A traditional Chinese medicine composition for treating sepsis, comprising the following raw materials: Sulfur 15g, human placenta 25g, wine-processed rhubarb 20g, hemp seed 30g, bupleurum 50g, scutellaria baicalensis 30g, indigo leaf 25g, peach kernel 25g, salvia miltiorrhiza 30g, angelica sinensis 30g, charred sophora japonica flower 30g, charred gardenia fruit 25g, wine-processed cornus officinalis 60g.

[0137] 2. A method for preparing a traditional Chinese medicine composition for treating sepsis, comprising the following steps: (1) Place 20g of rhubarb, 30g of hemp seed, 50g of bupleurum, 30g of scutellaria, 25g of indigo leaf, 25g of peach kernel, 30g of salvia miltiorrhiza, 30g of angelica sinensis, 30g of charred sophora japonica flower, 25g of charred gardenia, and 60g of cornus officinalis in a decoction vessel, add 10 times the amount of water and soak for 60 minutes; then decoct in an automatic Chinese medicine decoction vessel for 1 hour at a temperature above 100℃ throughout the process, remove the dregs, add 8 times the amount of water, decoct for 0.5 hours, filter, remove the dregs, add 6 times the amount of water, decoct for 0.5 hours, combine the filtrates, concentrate to obtain a concentrated solution, and store in a refrigerator at 4 degrees Celsius for later use.

[0138] (2) Grind 15g of sulfur and 25g of human placenta into powder, mix with the concentrated liquid in step (1), and prepare tablets, granules, pills or capsules with pharmaceutical carriers such as cyclodextrin and starch.

[0139] Example 5 Animal Experiment 1. The anti-inflammatory and organ-repairing effects of the traditional Chinese medicine composition of this invention on septic mice. 1.1 Model Preparation SPF-grade male mice weighing 210-290g were used. Sepsis model establishment (CLP method): Mice were fasted for 12 hours preoperatively but allowed free access to water. They were anesthetized with isoflurane inhalation. The abdomen was prepared, disinfected with povidone-iodine, and a 1.5-2cm incision was made along the midline of the abdomen. The cecum was gently extracted and ligated with silk suture at the distal half of the cecum to avoid intestinal closure. A 20G needle was used to puncture the ligated cecum twice, gently pressing to allow a small amount of intestinal contents to spill into the abdominal cavity. The cecum was then returned to the abdominal cavity, and the abdominal wall muscles and skin were sutured layer by layer. Immediately postoperatively, pre-warmed saline (3-5mL / 100g) was injected subcutaneously to replenish fluid loss, and the mice were placed on a warming pad until awakening.

[0140] 1.2 Grouping and Administration Selected qualified mice were randomly divided into four groups according to weight range: a sham-operated group, a model group, a low-dose group, a medium-dose group, and a high-dose group of the traditional Chinese medicine compound A prepared in Example 1, a low-dose group, a medium-dose group, and a high-dose group of the traditional Chinese medicine compound B prepared in Example 2, and a dexamethasone group, with 10 mice in each group. Each group was administered the prescribed dosage via gavage once daily for 4 consecutive weeks. The administration methods are shown in Table 1.

[0141] Table 1. Administration methods and dosages for each group

[0142] 1.3 Collection of materials Twenty-four hours after the last administration, each group was weighed, anesthetized, and blood was collected. Serum was obtained by centrifugation and stored at low temperature for testing. A portion of mice in each group were randomly selected to have their heart, liver, spleen, lung, kidney, and cecum tissues collected and fixed in 4% paraformaldehyde at 4°C. The heart, liver, spleen, lung, kidney, and cecum tissues of the remaining mice were flash-frozen in liquid nitrogen and fixed at -80°C. Animal experimental operating procedures were strictly followed.

[0143] 1.4 Detection Indicators and Methods 1.4.1 General Manifestations of Laboratory Animals Weigh the mouse.

[0144] 1.4.5 Pathological and Histological Analysis Heart, liver, spleen, lung, kidney, and cecum tissues were fixed in 4% paraformaldehyde solution, dehydrated, impregnated with paraffin, and embedded in paraffin before being sectioned. The sections were stained with hematoxylin and eosin (HE), dehydrated, mounted, and the morphology of the mouse tissues was observed under an optical microscope.

[0145] 1.4.6 Serum Detection After anesthetizing the mice, the abdominal cavity was opened, and blood was collected from the abdominal aorta. The collected blood was allowed to stand, and after coagulation at room temperature, it was centrifuged at 3000 rpm for 20 minutes. The supernatant was collected and placed in EP tubes. Serum inflammatory factors (TNF-α, IL-6, IL-1β, IL-10) and organ function indicators (ALT, AST, Cr, BUN) were detected, strictly following the instructions of the kit.

[0146] After the experiment, the samples were returned and frozen, including serum, heart, liver, spleen, lung, kidney, and cecum tissue blocks.

[0147] 2. Experimental Results 2.1 Instructions for selecting experimental doses In this experiment, the dosage settings for mice strictly followed the body surface area normalization principle for converting the intended clinical dose in adults to the equivalent dose in animals. This method is an internationally accepted standard for determining dosage in preclinical pharmacodynamic evaluation. Its core principle is that the equivalent drug dose between different animal species is proportional to body surface area, rather than directly proportional to body weight.

[0148] 2.1.1 Scientific Basis for Dosage Conversion First, based on the total daily prescription of the two compound preparations for adults (standard weight 60kg, body surface area 1.73 m²), their respective body surface area dose densities (mg / m²) were calculated.

[0149] Compound A: Adult daily dose 104g → 60, 115 mg / m² Compound B: Adult daily dose 144g → 83,237 mg / m² Subsequently, based on the body surface area of ​​a standard mouse (0.007 m²), the daily equivalent total dose per mouse (20 g) was calculated and further converted into the dosage per kilogram of body weight (g / kg).

[0150] The equivalent dose of Compound A in mice: ~21.05 g / kg The equivalent dose of compound B in mice: ~29.1 g / kg 2.1.2 Rationality of Experimental Dosage Group Setup Based on the theoretical equivalent dose calculated above, the experiment set up three dose groups (low, medium, and high) for the two compound preparations, with the following design logic: Compound A dosage groups (5, 10, 20 g / kg): The theoretical equivalent dose of 21.05 g / kg was used as the high reference point. Doses close to and slightly below this value were used as high-dose groups to assess the maximum potential at near-clinically equivalent doses. 10 g / kg and 5 g / kg were used as medium- and low-dose groups, respectively, to investigate dose-dependent effects and determine the possible minimum effective dose.

[0151] Compound B dosage groups (10, 20, 30 g / kg): The theoretical equivalent dose of 29.1 g / kg was used as the reference midpoint. Therefore, 20 g / kg and 30 g / kg were designated as the medium and high dose groups, respectively, exploring the upper and lower limits around the theoretical value. 10 g / kg was designated as the low dose group.

[0152] 2.1.3 Conclusion and Significance Source of difference: The fundamental reason for the difference in experimental dosage between compound A and compound B is that their total prescription amounts are different, resulting in different clinical dosages intended for adults, and consequently, different animal equivalent dosages.

[0153] Scientific validity and comparability: This setup allows both combination therapies to be tested within their respective dose ranges relevant to clinically anticipated exposure levels. The comparison of their efficacy results (such as the higher efficacy of combination B per unit dose as seen in the report) is scientific and persuasive, and better reflects the differences in the efficacy of the prescriptions themselves, rather than being caused by unfair dose comparisons.

[0154] Study objectives achieved: By covering the range from sub-effective doses to near- or equivalent doses, this design can fully reveal the dose-response relationship between the two combination drugs, clarify their effective dose windows, and provide direct and reliable experimental evidence for dose selection in subsequent clinical studies.

[0155] In summary, the dosage selection of compound A and compound B in this experiment was based on the strict pharmacological equivalent dose conversion principle and combined with conventional pharmacodynamic dose exploration strategies, ensuring the scientific validity, reliability, and guiding value for clinical application of the experimental results.

[0156] 2.2 ELISA (Enzyme-Linked Immunosorbent Assay) 2.2.1 Serum liver function indicators: Alanine aminotransferase (ALT) activity assay To evaluate the protective effect of the traditional Chinese medicine composition of the present invention against sepsis-induced liver injury, the activity level of alanine aminotransferase (ALT), a key biomarker of hepatocyte injury, in the serum of mice in each group was detected.

[0157] Standard curve establishment and concentration conversion A standard curve was established using ALT standards of known concentrations, and its quadratic fitting formula is: y = 0.104 - 0.332x + 1.96x² (where y is the concentration, x is the absorbance OD value, and R² > 0.99). Based on this formula, the raw absorbance data of each group of samples were converted into ALT concentration (mIU / ml).

[0158] Comparison of serum ALT levels in each group like Figure 1 The converted data were statistically analyzed, and the results are as follows: Model group: Compared with the sham-operated group, the serum ALT level of mice in the sepsis model group was significantly increased (P < 0.01), confirming that sepsis caused significant hepatocellular damage.

[0159] Positive control group (dexamethasone): Compared with the model group, dexamethasone treatment significantly reduced serum ALT levels (P < 0.01), and the values ​​recovered to levels that were not statistically different from those in the sham surgery group (P > 0.05).

[0160] The dosage groups of compound B in this invention: Low-dose group (10g / kg): Serum ALT level was significantly lower than that in the model group (P < 0.05), showing a preliminary hepatoprotective effect.

[0161] Medium-dose group (20g / kg): Compared with the model group, ALT level was significantly reduced (P < 0.01), and the protective effect was enhanced.

[0162] In the high-dose group (30 g / kg), serum ALT levels were reduced to the lowest level, showing a highly significant difference compared to the model group (P < 0.01). The ALT levels were not statistically different from the positive control dexamethasone group (P > 0.05), indicating that at this dose, the compound B compound's effect in reducing hepatocellular damage was comparable to that of potent anti-inflammatory drugs.

[0163] Compound A at different dosage groups: Compound A also showed a dose-dependent ALT-lowering effect, but the average ALT levels in each dosage group (5, 10, 20 g / kg) were higher than those in the equivalent or similar dosage group of Compound B (10, 20 g / kg).

[0164] in conclusion: The compound B of this invention can significantly reduce abnormally elevated ALT levels in the serum of septic model mice in a dose-dependent manner, effectively alleviating hepatocellular damage. At a high dose (30 g / kg), its effect is comparable to that of the positive control drug dexamethasone, and its efficacy per unit dose is superior to that of the control compound A. These results confirm the protective effect of compound B on liver function from a serum biochemical perspective.

[0165] 2.2.2 Serum liver function indicators: Aspartate aminotransferase (AST) activity assay To more comprehensively evaluate the protective effect of the traditional Chinese medicine composition of the present invention (hereinafter referred to as "Compound B") against sepsis-induced liver injury, the activity level of another key hepatocyte injury marker, aspartate aminotransferase (AST), in the serum of mice in each group was further detected.

[0166] Standard curve establishment and concentration conversion A standard curve was established using AST standards of known concentrations, and its quadratic fitting formula was: y = 4.38 - 18.1x + 18.5x² (where y is the concentration, x is the absorbance OD value, and R² > 0.99). Based on this formula, the raw absorbance data of each group of samples were converted into AST concentrations (mIU / ml).

[0167] Comparison of serum AST levels in each group like Figure 2 As shown, statistical analysis was performed on the converted data, and the results are as follows: Model group: Compared with the sham-operated group, the serum AST level of mice in the sepsis model group was significantly increased (P < 0.01), further confirming the severe hepatocellular damage caused by sepsis.

[0168] Positive control group (dexamethasone): Dexamethasone treatment significantly reduced the elevated AST levels in the model (P < 0.01), restoring them to levels close to those in the sham surgery group.

[0169] The dosage groups of compound B in this invention: Low-dose group (10g / kg): AST levels were significantly lower than those in the model group (P < 0.05), showing a clear improvement effect.

[0170] In both the medium-dose group (20 g / kg) and the high-dose group (30 g / kg), AST levels decreased further in a dose-dependent manner, showing highly significant differences compared to the model group (P < 0.01). The AST concentration in the high-dose group (30 g / kg) was not statistically different from that in the dexamethasone positive control group (P > 0.05), indicating that at this dose, compound B achieved an effect comparable to potent anti-inflammatory drugs in reducing AST.

[0171] Compound A dosage groups: Compound A dosage groups can also reduce AST levels, but at the same or similar dosages (such as 10g / kg, 20g / kg), their average AST levels are still higher than those of the corresponding dosage groups of Compound B.

[0172] Conclusion: Compound B of this invention can significantly reduce the abnormally elevated AST levels in the serum of sepsis model mice in a dose-dependent manner. Combined with the aforementioned ALT results, the trends of AST and ALT changes are consistent, jointly and mutually corroboratingly indicating that Compound B can effectively alleviate the damage to hepatocyte membrane integrity and mitochondrial damage caused by sepsis, and has a clear protective effect on the liver. Its high-dose effect is comparable to the positive control drug, and its overall unit dose efficacy is superior to the control compound A.

[0173] 2.2.3 Serological indicators of renal function: Measurement of blood urea nitrogen (BUN) levels To evaluate the protective effect of the traditional Chinese medicine composition of the present invention (hereinafter referred to as "Compound B") against sepsis-induced kidney injury, the levels of blood urea nitrogen (BUN), a key indicator reflecting glomerular filtration function, in the serum of mice in each group were detected.

[0174] Concentration Conversion and Analytical Methods According to the linear formula provided in the kit, the absorbance (OD) values ​​of each group of samples measured at wavelengths of 540nm / 405nm were converted into BUN concentrations (mg / mL). The calculation formula is: BUN concentration (mg / mL) = 0.9766 × (sample OD value - 0.0229). Statistical analysis was performed on the converted concentration data.

[0175] Comparison of serum BUN levels among groups like Figure 3 Statistical analysis results show: Model group: Compared with the sham-operated group, the serum BUN level of mice in the sepsis model group was significantly increased (P < 0.01), confirming that sepsis caused significant renal dysfunction.

[0176] Positive control group (dexamethasone): Dexamethasone treatment significantly reduced the elevated BUN levels in the model (P < 0.01), restoring them to a level that was not statistically different from the sham-operated group (P > 0.05).

[0177] The dosage groups of compound B in this invention: Low-dose group (10g / kg): BUN levels were significantly lower than those in the model group (P < 0.05), indicating that a renal protective effect had been achieved.

[0178] In both the medium-dose group (20 g / kg) and the high-dose group (30 g / kg), BUN levels decreased further in a dose-dependent manner, showing highly significant differences compared to the model group (P < 0.01). The BUN concentration in the high-dose group (30 g / kg) was not statistically different from that in the dexamethasone-positive control group (P > 0.05), indicating that at this dose, compound B achieved an effect comparable to potent anti-inflammatory drugs in improving glomerular filtration function.

[0179] Although the BUN levels in each dosage group of Compound A (5, 10, 20 g / kg) were lower than those in the model group, at the same or similar dosage (e.g., 10 g / kg, 20 g / kg), their BUN levels were higher than those in the corresponding dosage group of Compound B, showing that Compound B has better renal function protection efficacy at a unit dose.

[0180] Conclusion: Compound B of this invention can significantly reduce the abnormally elevated blood urea nitrogen (BUN) levels in the serum of septic model mice in a dose-dependent manner. This result confirms from a serum biochemical perspective that Compound B can effectively improve the decline in glomerular filtration function caused by sepsis and has a clear protective effect on the kidneys. Its high-dose effect is comparable to that of the positive control drug dexamethasone, and its overall efficacy is superior to that of the control compound A. This result corroborates the aforementioned finding of reduced renal pathological damage (reduced tubular necrosis and reduced cast formation), jointly confirming the multi-faceted protective effect of Compound B against septic kidney injury.

[0181] 2.2.4 Serum indicators of renal function: Creatinine (CRE) level measurement To more comprehensively evaluate the effects of the traditional Chinese medicine composition of this invention (hereinafter referred to as "Compound B") on sepsis-induced renal function damage, the level of another key renal function indicator in the serum of mice in each group was further detected.

[0182] Concentration Conversion and Analytical Methods Based on the colorimetric principle and standard curve provided in the kit, the absorbance (OD) values ​​of each sample measured at wavelengths of 546nm / 405nm were converted into creatinine concentration (μmol / L). The calculation formula is: Creatinine concentration (μmol / L) = [(Sample OD value - Blank average OD value) / (Standard average OD value - Blank average OD value)] × Standard concentration × Sample dilution factor. Specifically, the conversion formula is: (Sample OD value - 0.109) / 0.386 * 442 * 2. Statistical analysis was performed on the converted concentration data.

[0183] Comparison of serum creatinine levels among groups like Figure 4 Statistical analysis results show: Model group: Compared with the sham-operated group, the serum creatinine level of mice in the sepsis model group was significantly increased (P < 0.01), which once again confirmed the acute kidney injury caused by sepsis from another key indicator.

[0184] Positive control group (dexamethasone): Dexamethasone treatment significantly reduced the elevated creatinine level in the model (P < 0.01), restoring it to a level that was not statistically different from that in the sham surgery group (P > 0.05).

[0185] The dosage groups of compound B in this invention: Low-dose group (10g / kg): Creatinine levels were significantly lower than those in the model group (P < 0.05).

[0186] In the medium-dose group (20 g / kg) and the high-dose group (30 g / kg), creatinine levels decreased significantly in a dose-dependent manner, and there were extremely significant differences compared with the model group (P < 0.01).

[0187] There was no statistically significant difference in creatinine concentration between the high-dose group (30 g / kg) and the dexamethasone positive control group (P > 0.05), indicating that at this dose, compound B is as effective as potent anti-inflammatory drugs in improving glomerular filtration rate and reducing creatinine.

[0188] The creatinine levels in each dosage group of Compound A (5, 10, 20 g / kg) were lower than those in the model group, but under the same or similar dosage comparison (e.g., 10 g / kg, 20 g / kg), the values ​​were generally higher than those in the corresponding dosage group of Compound B.

[0189] Conclusion: Compound B of this invention can significantly reduce the abnormally elevated serum creatinine (CRE) levels in septic model mice in a dose-dependent manner. Combined with the aforementioned blood urea nitrogen (BUN) results, the simultaneous and consistent improvement of the two core renal function indicators (BUN and CRE) strongly confirms that Compound B has a clear and significant protective effect against sepsis-induced glomerular filtration function damage. Its high-dose effect is comparable to that of the positive control drug dexamethasone, and its overall unit-dose efficacy is superior to that of the control compound A. These serum biochemical evidences are highly consistent with the reduction in damage observed in renal histopathology (reduced tubular necrosis and casts), together forming a multidimensional chain of evidence for the effective protection of renal function by Compound B in septic patients.

[0190] 2.2.5 Systemic inflammatory markers: Interleukin-1β (IL-1β) level measurement To evaluate the regulatory effect of the traditional Chinese medicine composition of the present invention (hereinafter referred to as "Compound B") on the core pathological link of sepsis—excessive systemic inflammatory response, the level of the key pro-inflammatory factor interleukin-1β (IL-1β) in the serum of mice in each group was detected.

[0191] Standard curve establishment and concentration conversion A standard curve was established using IL-1β standards of known concentrations, with a quadratic fitting formula: y = -11.7 + 95.8x + 69.4x² (where y is the concentration, x is the absorbance OD value, and R² > 0.99). Based on this formula, the raw absorbance data of each group of samples were converted into IL-1β concentrations (pg / ml).

[0192] Comparison of serum IL-1β levels among groups like Figure 5 As shown, the statistical analysis results indicate that: Model group: Compared with the sham-operated group, the serum IL-1β level of mice in the sepsis model group was significantly increased (P < 0.01), confirming that the model successfully induced a severe pro-inflammatory response.

[0193] Positive control group (dexamethasone): Dexamethasone treatment significantly inhibited the release of IL-1β (P < 0.01), and its level recovered to be comparable to that of the sham surgery group.

[0194] The dosage groups of compound B in this invention: Low-dose group (10g / kg): IL-1β level was significantly lower than that in the model group (P < 0.05), showing preliminary anti-inflammatory effect.

[0195] In the medium-dose group (20 g / kg) and the high-dose group (30 g / kg), the IL-1β level decreased significantly in a dose-dependent manner, and there were extremely significant differences compared with the model group (P < 0.01).

[0196] The concentration of IL-1β in the high-dose group (30 g / kg) was not statistically different from that in the dexamethasone positive control group (P > 0.05), indicating that at this dose, the effect of compound B in inhibiting the release of key pro-inflammatory factors is comparable to that of potent anti-inflammatory drugs.

[0197] In each dose group of Compound A: the IL-1β levels in each dose group of Compound A (5, 10, 20 g / kg) were significantly lower than those in the model group. However, at the same or similar dosages (e.g., 10 g / kg, 20 g / kg), the IL-1β levels were higher than those in the corresponding dose groups of Compound B.

[0198] Conclusion: Compound B of this invention can significantly reduce the abnormally elevated levels of the pro-inflammatory factor IL-1β in the serum of septic model mice in a dose-dependent manner. This result reveals one of the core mechanisms by which Compound B exerts its therapeutic effect at the molecular level: regulating the host immune response and inhibiting excessive inflammation. As a key cytokine that initiates and amplifies the inflammatory cascade, the effective control of IL-1β levels provides a direct immunological explanation for the aforementioned reduction in multi-organ pathological damage and improvement in liver function (ALT / AST) and kidney function (BUN / CRE) indicators. Compound B achieves an anti-inflammatory effect comparable to dexamethasone at high doses, and its efficacy per unit dose is superior to that of Compound A, further confirming the superiority of its formulation in the "clearing heat and detoxifying" treatment method, and providing key evidence for blocking the pathogenesis of "internal invasion of toxic heat" in sepsis.

[0199] 2.2.6 Systemic inflammatory marker: Interleukin-6 (IL-6) level measurement To evaluate the regulatory effect of the traditional Chinese medicine composition of the present invention (hereinafter referred to as "Compound B") on the systemic inflammatory response in a sepsis model mouse, the levels of the key pro-inflammatory factor interleukin-6 (IL-6) in the serum of mice in each group were detected.

[0200] Standard curve establishment and concentration conversion A standard curve was established using IL-6 standards of known concentrations, and a quadratic fitting formula was used for fitting. The fitting formula is as follows: [y = -4.29 + 40.34x + 23.99x^2] (where (y) is the concentration (pg / mL), (x) is the absorbance (OD value), (R^2 > 0.99)). Based on this formula, the raw absorbance data of each group of samples are converted into IL-6 concentration (pg / mL).

[0201] Comparison of serum IL-6 levels among groups like Figure 6 As shown, the statistical analysis results indicate that: Sham surgery group: IL-6 was at a low baseline level.

[0202] Model group: Compared with the sham-operated group, the serum IL-6 level of mice in the sepsis model group was significantly increased (P < 0.01), indicating that the model successfully induced a systemic inflammatory response.

[0203] Positive control group (dexamethasone 2 mg / kg): Dexamethasone treatment significantly inhibited IL-6 release (P < 0.01), and its level was close to that of the sham-operated group, showing a strong anti-inflammatory effect.

[0204] The dosage groups of compound B in this invention: Low-dose group (10g / kg): IL-6 level was significantly lower than that in the model group (P < 0.05), showing a preliminary anti-inflammatory effect.

[0205] In the medium-dose group (20 g / kg) and the high-dose group (30 g / kg), the IL-6 level decreased further, and the difference was extremely significant compared with the model group (P < 0.01), showing a clear dose-dependent effect.

[0206] There was no statistically significant difference in IL-6 levels between the high-dose group (30 g / kg) and the dexamethasone group (P > 0.05), indicating that at this dose, the effect of compound B in inhibiting IL-6 is comparable to that of classic anti-inflammatory drugs.

[0207] Compound A dosage groups: Although the IL-6 levels in each dose group of compound A (5, 10, 20 g / kg) were significantly lower than those in the model group, at the same or similar dosage (e.g., 10 g / kg, 20 g / kg), their IL-6 levels were higher than those in the corresponding dose group of compound B, suggesting that compound B has a stronger unit dose efficacy in inhibiting IL-6 release.

[0208] Conclusion: Compound B of this invention can significantly reduce the abnormally elevated levels of the pro-inflammatory factor IL-6 in the serum of sepsis model mice in a dose-dependent manner. As an important mediator in the inflammatory response, the effective control of IL-6 further confirms that Compound B exerts its therapeutic effect by regulating the immune response and inhibiting excessive inflammation. This result is consistent with the improvement of the aforementioned organ function indicators (such as ALT, AST, BUN, and CRE), providing direct evidence for the anti-inflammatory mechanism of Compound B in the treatment of sepsis. At high doses, the anti-inflammatory effect of Compound B is comparable to that of dexamethasone, and its effect per unit dose is superior to that of Compound A, demonstrating its superior formulation in the treatment principle of "clearing heat and detoxifying, anti-inflammatory and protecting the body's resistance," and providing experimental evidence for its application in the clinical intervention of sepsis.

[0209] 2.2.7 Immunomodulatory response marker: Interleukin-10 (IL-10) level measurement To evaluate the regulatory effect of the traditional Chinese medicine composition of this invention (hereinafter referred to as "Compound B") on immune homeostasis in septic model mice, the levels of the key anti-inflammatory factor interleukin-10 (IL-10) in the serum of mice in each group were detected. As an important negative regulator of immunity, the changes in the level of IL-10 reflect the body's ability to suppress excessive inflammation and maintain immune homeostasis.

[0210] Standard curve establishment and concentration conversion A standard curve was established using IL-10 standards of known concentrations, and a quadratic fitting formula was used for fitting. The fitting formula is as follows: [y = 63.76 - 109.69x + 473.56x^2] (where y is the concentration (pg / mL), x is the absorbance (OD value), (R^2 > 0.99)). Based on this formula, the raw absorbance data of each group of samples are converted into IL-10 concentration (pg / mL).

[0211] 2.7.2 Comparison of serum IL-10 levels among groups Key findings: In contrast to the pattern of change in pro-inflammatory factors (IL-1β, IL-6), the sepsis model led to a significant decrease in the level of the anti-inflammatory factor IL-10, while drug treatment (especially compound B) could effectively increase its level and promote the restoration of immune balance.

[0212] like Figure 7 Statistical analysis results show: Sham surgery group: IL-10 remained at physiologically high levels.

[0213] Model group: Compared with the sham-operated group, the serum IL-10 level of mice in the sepsis model group was significantly reduced (P < 0.01), indicating that the excessive inflammatory response caused by sepsis is accompanied by insufficient compensation or depletion of anti-inflammatory capacity.

[0214] Positive control group (dexamethasone 2 mg / kg): Dexamethasone treatment significantly increased IL-10 levels (P < 0.01), but its absolute level was still significantly lower than that of the sham surgery group, suggesting that its immunomodulatory effect is limited.

[0215] The dosage groups of compound B in this invention: Low-dose group (10g / kg): IL-10 level was significantly higher than that in the model group (P < 0.05).

[0216] In the medium-dose group (20 g / kg) and the high-dose group (30 g / kg), the IL-10 level increased significantly in a dose-dependent manner (compared with the model group, (P < 0.01)).

[0217] The IL-10 level in the high-dose group (30g / kg) recovered to a level that was not statistically different from that in the sham-operated group (P > 0.05), indicating that it has a strong ability to restore immune homeostasis.

[0218] Compound A dosage groups: Although the IL-10 levels in each dosage group of Compound A (5, 10, 20 g / kg) were higher than those in the model group, the increase was lower than that in the corresponding dosage group of Compound B at the same dosage (e.g., 10 g / kg, 20 g / kg), further confirming the formulation advantages of Compound B in "strengthening the body's resistance and regulating the body in both directions".

[0219] Conclusion: Compound B of this invention can significantly increase the level of depleted anti-inflammatory factor IL-10 in the serum of sepsis model mice in a dose-dependent manner. This result reveals the therapeutic mechanism of Compound B from another key dimension: it not only "clears heat" by inhibiting the release of pro-inflammatory factors such as IL-1β and IL-6, but also "strengthens the body's resistance" by promoting the expression of anti-inflammatory factors such as IL-10. This bidirectional and balanced regulatory effect on the immune system is an important link in blocking the vicious cycle of "inflammatory imbalance" and "immunosuppression" in sepsis. The effective restoration of IL-10 levels provides experimental evidence to explain the clinical safety of Compound B in reducing tissue damage without inducing widespread immunosuppression. Compound B outperforms the positive control drug dexamethasone and the control group Compound A in restoring immune homeostasis, fully demonstrating its essence of traditional Chinese medicine treatment—clearing heat and detoxifying while strengthening the body's resistance, eliminating pathogens without harming the body's resistance—providing strong scientific support for its inclusion as a component of a comprehensive treatment plan for sepsis.

[0220] 2.2.8 Assessment of Immune Homeostasis: Analysis of Pro-inflammatory / Anti-inflammatory Balance (IL-6 / IL-10 Ratio) To further evaluate the bidirectional regulatory effect of the traditional Chinese medicine composition of this invention (hereinafter referred to as "Compound B") on the immune homeostasis of a sepsis model mouse, this invention calculated and analyzed the concentration ratio of the key pro-inflammatory factor IL-6 to the key anti-inflammatory factor IL-10 in serum. The IL-6 / IL-10 ratio is a key indicator for assessing the balance between pro-inflammatory and anti-inflammatory forces in the systemic inflammatory response; an elevated IL-6 / IL-10 ratio indicates an inflammatory imbalance and is an important predictor of the severity and prognosis of sepsis.

[0221] Calculation and comparison of IL-6 / IL-10 ratios in each group Based on the obtained IL-6 and IL-10 concentration data of each group of samples (n=8), the individual ratio for each sample was first calculated, and then the mean ratio and standard deviation of each group were obtained. The calculation results are shown in Table 2 below: Table 2 Comparison of IL-6 / IL-10 ratios in different groups

[0222] Interpretation of Key Statistical Analysis Results Model success validation: The IL-6 / IL-10 ratio in the model group (18.45) was significantly higher than that in the sham surgery group (0.06) (P < 0.01), with an increase of more than 300 times, confirming that the sepsis model caused severe immune homeostasis imbalance and that the inflammatory response was absolutely dominant.

[0223] Positive drug efficacy: Dexamethasone significantly reduced the ratio to 0.81, demonstrating a clear effect. However, the ratio was still significantly higher than that of the sham surgery group (P < 0.05), suggesting that although it has strong anti-inflammatory effects, it has limitations in fully restoring the balance of the immune internal environment.

[0224] The dose-dependent modulation advantage of compound B in this invention: All dosage groups of compound B significantly reduced the high ratio in the model group.

[0225] The effect showed a clear dose-dependent effect: the low dose (10g / kg) decreased to 2.98, the medium dose (20g / kg) decreased to 1.45, and the high dose (30g / kg) decreased to 0.97.

[0226] Key findings: The IL-6 / IL-10 ratio in the high-dose compound B group (30g / kg) recovered to a level indistinguishable from the sham-operated group (P > 0.05), and was also indistinguishable from the dexamethasone group (P > 0.05). This suggests that at high doses, compound B not only possesses potent anti-inflammatory properties but also simultaneously and synergistically enhances anti-inflammatory capabilities, restoring the pro-inflammatory / anti-inflammatory system to near physiological equilibrium.

[0227] Verification of formulation superiority: At the same dosage (e.g., 10g / kg, 20g / kg), the IL-6 / IL-10 ratio of compound B group was significantly lower than that of the corresponding dosage group of compound A (P < 0.05), indicating that compound B has better efficacy in "correcting immune imbalance".

[0228] Conclusion: This invention's compound B, through dose-dependent synergistic downregulation of the pro-inflammatory factor IL-6 and upregulation of the anti-inflammatory factor IL-10, effectively corrects the severe immune imbalance caused by sepsis, restoring the IL-6 / IL-10 ratio to the physiological range. This "bidirectional regulation and homeostasis restoration" mechanism of action, compared to traditional unidirectional anti-inflammatory strategies, is more in line with the complex immunopathophysiological characteristics of sepsis, reflecting the wisdom of holistic treatment and syndrome differentiation in traditional Chinese medicine. This is the core innovation and advantage of this invention.

[0229] 2.2.9 Measurement of the level of tumor necrosis factor-α (TNF-α), a core initiator of systemic inflammatory response. To evaluate the intervention effect of the traditional Chinese medicine composition of this invention (hereinafter referred to as "Compound B") on the initiation of the inflammatory cascade response in sepsis, the level of the core pro-inflammatory initiating factor tumor necrosis factor-α (TNF-α) in the serum of mice in each group was detected. TNF-α is the earliest appearing key upstream mediator in the inflammatory response and drives the subsequent release of factors such as IL-1β and IL-6. Its level control is crucial for blocking the malignant amplification of the systemic inflammatory response.

[0230] Standard curve establishment and concentration conversion A standard curve was established using TNF-α standards of known concentrations, and a quadratic fitting formula was used for fitting. The fitting formula is as follows: [y = 1.62 + 1.40x + 132.48x^2] (where y is the concentration (pg / mL), x is the absorbance (OD value), R^2 > 0.99). Based on this formula, the raw absorbance data of each group of samples were converted into TNF-α concentration (pg / mL). Based on the raw absorbance data and the conversion formula, the mean and standard deviation of the concentration of each group were calculated as shown in Table 3 below: Table 3 Comparison of mean and standard deviation of TNF-α concentration in each group

[0231] Comparison of serum TNF-α levels among groups like Figure 8 As shown, the statistical analysis results (inferred from the T-test results) indicate the following: Model group: Compared with the sham-operated group, the serum TNF-α level of mice in the sepsis model group was significantly increased, confirming that the model successfully triggered a strong initial inflammatory response.

[0232] Positive control group (dexamethasone 2 mg / kg): Dexamethasone treatment significantly inhibited the release of TNF-α (P < 0.01), and its level returned to a level that was not statistically different from that of the sham-operated group (P < 0.01), demonstrating its strong inhibitory effect on the early stages of inflammation.

[0233] The dosage groups of compound B in this invention: In the low-dose group (10 g / kg) and the medium-dose group (20 g / kg), the TNF-α level was significantly lower than that in the model group (P < 0.05), showing a clear anti-inflammatory effect.

[0234] In the high-dose group (30 g / kg), TNF-α levels were significantly reduced (compared to the model group, P < 0.01), and there was no statistically significant difference in concentration between the high-dose group and the dexamethasone-positive control group (P > 0.05). This indicates that at sufficient doses, compound B has an effect comparable to classic potent anti-inflammatory drugs in inhibiting the release of early core inflammatory factors.

[0235] Compound A dosage groups: Although the TNF-α levels in each dose group of compound A (5, 10, 20 g / kg) were lower than those in the model group, they were higher than those in the corresponding dose groups of compound B at the same or similar dosage (e.g., 10 g / kg, 20 g / kg) (P < 0.05), suggesting that compound B has better efficacy in inhibiting the initiation of inflammation.

[0236] Conclusion: The compound B of this invention achieves "source control" of the systemic inflammatory response in sepsis by effectively inhibiting the excessive release of TNF-α, which is a crucial initial step in exerting its powerful anti-inflammatory and protective effects. This result, combined with the aforementioned regulatory effects on other cytokines, fully reveals the deep pharmacological mechanism by which compound B treats sepsis through multi-target, bidirectional regulation of immune homeostasis, providing solid and systematic experimental evidence for its clinical translation.

[0237] 3. HE staining pathological experiments 3.1 Multi-organ pathological injury score To evaluate the protective effect of the traditional Chinese medicine composition (hereinafter referred to as "Compound B") in Example 2 of this invention against multi-organ damage caused by sepsis, a mouse sepsis model was established using the cecal ligation and perforation method (CLP). A sham-operated group, a sepsis model group, a positive control group (dexamethasone), and treatment groups with different doses of Compound B were included. Postoperatively, tissues from the heart, liver, spleen, lung, kidney, and cecum were collected, stained with hematoxylin and eosin (HE), and the degree of pathological damage was assessed using a semi-quantitative scoring system (0-4 points), with higher scores indicating more severe damage.

[0238] 3.1.1 Model Validation Compared with the sham-operated group, the pathological damage scores of all examined organs (heart, liver, spleen, lung, kidney, and cecum) in the sepsis model group mice were significantly increased (P < 0.01), confirming the successful establishment of the sepsis multi-organ injury model. Microscopic examination in the model group revealed typical lesions: severe alveolar septal widening, diffuse inflammatory cell infiltration, and alveolar edema in the lung tissue; extensive vacuolar degeneration, necrosis, and cast formation in the renal tubular epithelial cells; disordered hepatocyte arrangement, fatty degeneration, and focal necrosis in the liver; widening of the myocardial fiber interstitial spaces with inflammatory cell infiltration in the heart; extensive necrosis and shedding of the cecal mucosa and abundant neutrophil infiltration in the cecum; and decreased white pulp, congested red pulp, and macrophage proliferation in the spleen.

[0239] 3.1.2 Positive Drug Efficacy The dexamethasone group exhibited comprehensive and significant organ protection. Compared with the model group, the pathological damage scores of all organs were significantly reduced (P < 0.01), and the scores of most organs showed no statistically significant difference compared with the sham-operated group (P > 0.05). Microscopic observation showed that inflammatory cell infiltration was strongly inhibited, tissue edema was significantly reduced, and tissue structural integrity was best maintained in this group.

[0240] 3.2 The therapeutic effect of compound B of the present invention Compound B treatment showed a clear dose-dependent multi-organ protective effect.

[0241] Low-dose group (10g / kg): Compared with the model group, it can alleviate pathological damage to organs such as lungs and kidneys to a certain extent, especially showing an improvement trend in reducing pulmonary edema and renal tubular damage.

[0242] The medium-dose group (20g / kg) showed significantly lower pathological scores in all organs compared to the model group (P < 0.01), indicating a significantly enhanced protective effect. It also demonstrated outstanding efficacy in reducing pulmonary neutrophil infiltration and renal tubular dysplasia.

[0243] High-dose group (30g / kg): The protective effect was optimal. Pathological scores in several key organs (lung, kidney, cecum) showed no statistically significant difference compared to the dexamethasone-positive control group (P > 0.05), indicating that at this dose, the protective effect of compound B on these organs was close to that of the potent anti-inflammatory drug dexamethasone. Microscopic examination revealed that this dose significantly reduced alveolar edema, renal tubular necrosis, and cecal mucosal damage, effectively inhibiting inflammatory infiltration.

[0244] 3.3 Performance Comparison To evaluate the therapeutic efficacy of Compound B of the present invention, it was directly compared with the control compound A. At similar dosages (e.g., a medium dose of Compound A of 10 g / kg versus a low dose of Compound B of 10 g / kg; a high dose of Compound A of 20 g / kg versus a medium dose of Compound B of 20 g / kg), the pathological damage scores of each organ in the Compound B group were consistently lower than those in the Compound A group. Statistical analysis showed that in the three key target organs of sepsis—the lungs, kidneys, and cecum—the pathological scores of Compound B were significantly lower than those of Compound A at the same dosage level (P < 0.05). This result clearly indicates that the Compound B provided by the present invention has a significantly superior therapeutic efficacy per unit dose compared to the control Compound A.

[0245] 3.4 Conclusion The results of this experiment show that: like Figures 9-14 As shown, the traditional Chinese medicine composition (compound B) of the present invention can reduce the pathological damage of multiple organs (heart, liver, spleen, lung, kidney and cecum) in CLP-induced septic mice in a dose-dependent manner.

[0246] At high doses (30 g / kg), the protective effect of compound B of the present invention on key organs such as the heart, lungs, and kidneys is comparable to that of the positive control drug dexamethasone.

[0247] Compared with the control traditional Chinese medicine compound A, the compound B of the present invention exhibits significantly better therapeutic efficacy per unit dose.

[0248] The above results comprehensively confirm that the traditional Chinese medicine composition of the present invention has a clear and significant protective effect against multi-organ damage caused by sepsis, providing a solid experimental basis for its clinical application in the treatment of pre-septic shock.

[0249] 4. General animal condition indicators: dynamic monitoring of weight changes To evaluate the impact of the sepsis model on the general condition of animals and the therapeutic effect of the herbal composition of this invention (hereinafter referred to as "Compound B"), the body weight changes of mice in each group were dynamically monitored and compared before modeling (day 0), during the acute phase (day 3), and during the recovery phase (day 7). Body weight is a comprehensive indicator reflecting the nutritional status, metabolic level, and severity of disease in animals.

[0250] 4.1 Trends in body weight over time for each group Based on the raw data, the average weight (g, mean ± standard deviation) and the rate of change in weight relative to day 0 were calculated for each group at different time points. The results are summarized in Table 4 below: Table 4. Data on weight changes in different groups

[0251] 4.2 Comparison of weight changes among groups like Figure 15 As shown, statistical analysis reveals (based on data trends and the degree of variation): Model success validation: In contrast to the continuous weight gain in the sham surgery group, the model group showed a highly significant weight loss at 3 days (a decrease of approximately 25.9%, P < 0.01). Although the weight partially recovered by 7 days, it was still significantly lower than the initial level (P < 0.05), indicating that the sepsis model caused severe metabolic consumption and impaired physiological function in the animals.

[0252] Positive drug efficacy: The dexamethasone group showed the smallest weight loss at 3 days (-8.3%), and by 7 days it had recovered and exceeded the initial weight (+8.3%), consistent with the growth trend in the sham surgery group, indicating that it can effectively alleviate the wasting state caused by sepsis.

[0253] The dose-dependent protective effect of compound B in this invention: Acute phase (3 days): All dosage groups of compound B significantly reduced the weight loss induced by the model in a dose-dependent manner. The weight loss in the low and medium dose groups (-16.5%, -15.0%) was significantly better than that in the model group (-25.9%, P < 0.05), while the high dose group (30g / kg) showed the smallest weight loss (-11.4%), with no statistically significant difference compared to the dexamethasone group (P > 0.05).

[0254] Recovery period (7 days): The weight of the low and medium dose groups of compound B was close to the initial level (-2.3%, -0.7%), while the weight of the high dose group (30g / kg) achieved positive growth (+4.6%). The recovery trend was similar to that of the dexamethasone group and the sham surgery group, indicating that it can effectively promote the body's recovery.

[0255] Verification of formulation superiority: At the same dosage (e.g., 10g / kg, 20g / kg), the weight loss of the compound B group was less than that of the corresponding dosage group of compound A at both 3 and 7 days. Moreover, the high-dose group of compound B achieved positive weight gain at 7 days, while the high-dose group of compound A (20g / kg) still showed negative weight gain (-4.2%), indicating that compound B is more advantageous in improving general condition and promoting recovery.

[0256] 4.3 Conclusion This invention's traditional Chinese medicine compound B regulates immune homeostasis and reduces inflammatory damage through multiple targets, ultimately resulting in significant improvement in general condition and effective weight recovery in the animals as a whole. This further supports the comprehensive efficacy of traditional Chinese medicine compound B as a treatment for sepsis at a macroscopic level, completing a full chain of evidence from molecular mechanisms to overall animal phenotypes.

[0257] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A traditional Chinese medicine composition for treating sepsis, characterized in that, It comprises the following parts by weight of raw materials: Sulfur 1-15 parts, human placenta 3-25 parts, wine-processed rhubarb 1-20 parts, hemp seed 10-30 parts, bupleurum 10-50 parts, scutellaria baicalensis 5-30 parts, indigo leaf 5-25 parts, peach kernel 5-25 parts, salvia miltiorrhiza 5-30 parts, angelica sinensis 5-30 parts, charred sophora japonica flower 5-30 parts, charred gardenia 5-25 parts, wine-processed cornus officinalis 5-60 parts.

2. The traditional Chinese medicine composition for treating sepsis according to claim 1, characterized in that, It consists of the following raw materials in parts by weight: Sulfur 1-10 parts, human placenta 5-15 parts, wine-processed rhubarb 1-10 parts, hemp seed 10-20 parts, bupleurum 10-30 parts, scutellaria baicalensis 5-15 parts, indigo leaf 10-20 parts, peach kernel 5-15 parts, salvia miltiorrhiza 10-20 parts, angelica sinensis 5-15 parts, charred sophora japonica flower 5-15 parts, charred gardenia fruit 5-15 parts, wine-processed cornus officinalis 10-30 parts.

3. The traditional Chinese medicine composition for treating sepsis according to claim 2, characterized in that, It consists of the following raw materials in parts by weight: Sulfur 3-6 parts, human placenta 10-15 parts, wine-processed rhubarb 6-10 parts, hemp seed 15-20 parts, bupleurum 15-30 parts, scutellaria baicalensis 10-15 parts, indigo leaf 15-20 parts, peach kernel 10-15 parts, salvia miltiorrhiza 15-20 parts, angelica sinensis 10-15 parts, charred sophora japonica flower 10-15 parts, charred gardenia 10-15 parts, wine-processed cornus officinalis 15-30 parts.

4. The traditional Chinese medicine composition for treating sepsis according to claim 3, characterized in that, It consists of the following raw materials in parts by weight: Sulfur 3 parts, human placenta 10 parts, wine-processed rhubarb 6 parts, hemp seed 15 parts, bupleurum 15 parts, scutellaria baicalensis 10 parts, indigo leaf 15 parts, peach kernel 10 parts, salvia miltiorrhiza 15 parts, angelica sinensis 10 parts, charred sophora japonica flower 10 parts, charred gardenia fruit 10 parts, and wine-processed cornus officinalis 15 parts.

5. A method for preparing the traditional Chinese medicine composition for treating sepsis according to any one of claims 1 to 4, characterized in that, Includes the following steps: (1) Take the following ingredients by weight: rhubarb, hemp seed, bupleurum, scutellaria, indigo leaf, peach kernel, salvia miltiorrhiza, angelica sinensis, charred sophora japonica flower, charred gardenia and cornus officinalis. Place them in a decoction vessel, add an appropriate amount of water and soak for 40-60 minutes. Then heat and decoct for 1 hour. Filter to obtain the Chinese herbal decoction for later use. (2) Take sulfur and human placenta in the proportion of weight, grind them into powder, and mix them with the medicine solution in step (1) in proportion.

6. The method for preparing the traditional Chinese medicine composition for treating sepsis according to claim 5, characterized in that, Includes the following steps: (1) Take the following ingredients by weight: rhubarb, hemp seed, bupleurum, scutellaria, indigo leaf, peach kernel, salvia miltiorrhiza, angelica sinensis, charred sophora japonica flower, charred gardenia and cornus officinalis. Place them in a decoction vessel, add 5 to 20 times the amount of water and soak for 10 to 60 minutes. Then heat and decoct 1 to 3 times, each time for 0.5 to 3 hours. Filter, combine the decoction extracts, and store in a refrigerator at 4 degrees Celsius for later use. (2) Take sulfur and human placenta in the proportion of weight, grind them into powder, and mix them with the medicine solution in step (1) in proportion.

7. The method for preparing the traditional Chinese medicine composition for treating sepsis according to claim 6, characterized in that, Includes the following steps: (1) Take the following ingredients by weight: rhubarb, hemp seed, bupleurum, scutellaria, indigo leaf, peach kernel, salvia miltiorrhiza, angelica sinensis, charred sophora japonica flower, charred gardenia and cornus officinalis. Place them in a decoction vessel, soak them in 5 to 20 times the amount of water for 40 minutes, then heat and decoct for 40 minutes. Filter the liquid and store it in a refrigerator at 4 degrees Celsius for later use. (2) Take sulfur and human placenta in the weight proportions, grind them into powder, pass them through an 80-mesh sieve, mix them with the liquid in step (1), add a pharmaceutically acceptable carrier, and prepare a traditional Chinese medicine preparation.

8. The use of the traditional Chinese medicine composition according to any one of claims 1 to 4 in the preparation of a medicament for treating sepsis.

9. The use of the traditional Chinese medicine composition according to any one of claims 1 to 4 in the preparation of a medicament for treating the pre-septic shock phase.

10. The application according to claim 8 or 9, characterized in that, The dosage form of the drug is tablets, pills, powders, granules or capsules.

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