Traditional chinese medicine composition for postoperative gas expulsion, preparation method therefor, and use thereof

Abstract

A traditional Chinese medicine composition for postoperative gas expulsion, a preparation method therefor, and use thereof. The composition is prepared from the following traditional Chinese medicine components: 15-45 parts by weight of stir-fried Raphani semen, 5-15 parts by weight of stir-fried Perillae fructus, 5-15 parts by weight of stir-fried Sinapis semen, 4.5-13.5 parts by weight of stir-fried Corneum gigeriae galli endothelium, 5-15 parts by weight of charred Crataegi fructus, 3-9 parts by weight of Citri reticulatae pericarpium, 15-45 parts by weight of Taraxaci herba, and 1.5-4.5 parts by weight of Glycyrrhizae radix et rhizoma. The present invention integrates the traditional Chinese medicine concept of "the lung and large intestine sharing an exterior-interior relationship". After a stir-frying process, the properties of Raphani semen, Sinapis semen, and Perillae fructus shift from ascending to descending. The entire formula primarily directs the internal Qi downward, facilitating the expulsion of postoperative intestinal gas. A formulation is prepared according to the optimal processing method for each of the components and made into formula granules, providing beneficial effects for the pharmaceutical composition of the postoperative gas expulsion formula.

Description

Postoperative ventilation traditional Chinese medicine composition and preparation method and application thereof TECHNICAL FIELD

[0001] The present application belongs to the technical field of traditional Chinese medicine composition, and particularly relates to a postoperative ventilation traditional Chinese medicine composition and a preparation method and application thereof. BACKGROUND

[0002] Postoperative ileus (POI) is a clinical phenomenon that the gastrointestinal motility function is temporarily inhibited after surgery, and it is characterized by impaired gastrointestinal peristalsis function, and can clinically manifest as postoperative nausea, vomiting, abdominal pain, and stop of exhaust and defecation. From the perspective of pathophysiology, POI can be divided into a short-duration neurogenic stage and a long-duration inflammatory stage. The neurogenic stage usually disappears within a few hours after surgery, while the inflammatory stage can last for several days. Therefore, we take the inflammatory stage as the main intervention stage. Generally, it recovers within 3 days, but it can also persist or relapse, in which case it is called prolonged postoperative ileus (PPOI). The pathogenesis of POI is not fully understood at present. It is currently believed that abdominal surgical procedures can cause damage to the intestinal mucosal barrier, intestinal flora imbalance and immune regulation abnormalities, and activation of inflammatory cells, all of which can cause increased intestinal mucosal permeability and thus cause persistent inflammatory response. This disease can have serious consequences, including intestinal nutritional deficiency in patients, prolonged hospitalization, excessive economic burden, and reduced quality of life. Despite the continuous innovation of traditional perioperative management methods and surgical techniques, the incidence and mortality rates have been declining, but the duration of postoperative ileus after major abdominal surgery (tumor) still lasts for 48h-74h. The occurrence of postoperative ileus is mainly caused by the body's stress response due to surgical stimulation, the use of anesthetic drugs, intraoperative hypothermia, intraoperative inappropriate fluid management, postoperative pain, and long-term inactivity of patients.

[0003] According to Traditional Chinese Medicine (TCM) theory, this disease falls under the categories of "intestinal obstruction," "gastrointestinal blockage," and "intestinal paralysis." "Intestinal paralysis" first appeared in the *Suwen* (Plain Questions) section on "Bi Syndrome," describing it as "a condition where one frequently drinks but cannot, experiences shortness of breath and difficulty breathing, and occasionally suffers from diarrhea." Surgery and anesthesia can cause stagnation of Qi and blood, disrupting the downward flow of the gastrointestinal tract and leading to abdominal distension. The six fu organs function by "transmitting and transforming substances without storing them," and gastrointestinal function relies on downward flow for proper digestion. After abdominal surgery, due to impaired gastric function, symptoms such as pain, vomiting, distension, and constipation commonly appear, indicating gastrointestinal obstruction. Furthermore, TCM believes that major surgeries often deplete vital energy and essence, leading to Qi and blood deficiency, further exacerbating postoperative intestinal paralysis. This disease significantly prolongs hospital stays and increases treatment costs, but its pathogenesis still requires further investigation. Traditional Chinese medicine has shown significant clinical efficacy in treating postoperative gastrointestinal dysfunction, not only assisting in early recovery of gastrointestinal function but also significantly reducing the occurrence of postoperative complications.

[0004] Traditional Chinese medicine (TCM) has significant advantages in postoperative analgesia, prevention and treatment of stress responses, and improvement of gastrointestinal symptoms. Its green, safe, and effective clinical advantages, as well as its unique preventive and therapeutic effects on postoperative paralytic ileus, are precisely what postoperative patients urgently need. However, relevant research in this field is still lacking.

[0005] The application of traditional Chinese medicine composition for postoperative ventilation in postoperative patients has shown that it can advance the ventilation time and has good clinical efficacy in treating postoperative gastrointestinal dysfunction. This can effectively prevent and treat postoperative paralytic ileus, improve gastrointestinal symptoms, shorten hospital stay, save treatment costs, and accelerate patient recovery. Summary of the Invention

[0006] Technical problem solved: In view of the above-mentioned technical problem, the present invention provides a traditional Chinese medicine composition for postoperative ventilation, its preparation method and application.

[0007] Technical solution: A postoperative ventilation traditional Chinese medicine composition, which is made from the following traditional Chinese medicine components in parts by weight: 15-45 parts of stir-fried radish seed, 5-15 parts of stir-fried perilla seed, 5-15 parts of stir-fried yellow mustard seed, 4.5-13.5 parts of stir-fried chicken gizzard lining, 5-15 parts of charred hawthorn, 3-9 parts of dried tangerine peel, 15-45 parts of dandelion, and 1.5-4.5 parts of licorice.

[0008] As one of the preferred options, the above-mentioned traditional Chinese medicine compound composition for accelerating postoperative ventilation is made from the following traditional Chinese medicine components in parts by weight: 15-30 parts of stir-fried radish seeds, 5-10 parts of stir-fried perilla seeds, 5-10 parts of stir-fried yellow mustard seeds, 4.5-9 parts of stir-fried chicken gizzard lining, 5-10 parts of charred hawthorn, 3-6 parts of dried tangerine peel, 15-30 parts of dandelion, and 1.5-3 parts of licorice.

[0009] As a further preferred solution, the above-mentioned Chinese medicinal compound composition for accelerating postoperative ventilation is made of the following Chinese medicinal components in parts by weight: fried Semen Raphani 30 parts, fried Fructus Perillae 10 parts, fried Semen Raphani 10 parts, fried Semen Galli Sonatii 9 parts, charred Fructus Crataegi 10 parts, Pericarpium Citri Reticulatae Viride 6 parts, Herba Taraxaci 30 parts, and Glycyrrhizae 3 parts.

[0010] The preparation method of the above-mentioned Chinese medicinal composition for postoperative ventilation comprises the following steps: taking fried Semen Raphani, fried Semen Raphani, fried Fructus Perillae, Herba Taraxaci, fried Semen Galli Sonatii, charred Fructus Crataegi, Pericarpium Citri Reticulatae Viride, and Glycyrrhizae in parts by weight, decocting twice with water, adding water in the first extraction in an amount of 12 times the volume of the decoction pieces, soaking for 35 minutes first, and heating and decocting for 40 minutes, filtering, and taking the filtrate; adding water in the second extraction in an amount of 6 times the volume of the decoction pieces, heating and decocting for 30 minutes, filtering, and taking the filtrate; combining the filtrates of the two extractions, concentrating under reduced pressure to a relative density of about 1.05-1.10 at 65°C, then centrifuging with a straight-tube high-speed centrifuge to obtain a post-centrifugation solution. Taking an appropriate amount of dextrin, placing it in a fluidized bed, and when the material temperature rises, starting to feed the post-centrifugation solution at a feeding speed of 80-150 r / min, and atomizing and drying to obtain the product.

[0011] The above-mentioned composition is used for preparing a medicament for treating long-term postoperative ileus.

[0012] The above-mentioned composition is used for preparing a medicament for postoperative ventilation.

[0013] Fried Semen Raphani has a mild taste, is pungent, sweet, and neutral, and is attributed to the lung, spleen, and stomach meridians. It is mainly used for food digestion and swelling relief, and for reducing qi and resolving phlegm. The main chemical components are sinapine, sinapine thiocyanate, and fatty oil, etc. Semen Raphani has the effects of food digestion and swelling relief, and for reducing qi and resolving phlegm, and is used for treating food stagnation, abdominal distention and pain, constipation, accumulation and diarrhea, and phlegm accumulation and cough, etc. Semen Raphani is a typical Chinese medicinal herb for “different treatment of raw and cooked, ascending of raw and descending of cooked” in Chinese medicinal processing. Fried Semen Raphani has the effect of food digestion and swelling relief due to its fried nature, and is a commonly used food digestion herb in clinical practice. Modern research shows that Semen Raphani can contract the smooth muscles of isolated stomach and duodenum, and is used for constipation and abdominal distention, etc. Semen Raphani mainly contains glucosinolates and sulfur-containing derivatives, alkaloids, flavonoids, fatty acids, volatile oils, and polysaccharides, etc. Glucosinolates are the characteristic components before and after processing of Semen Raphani, among which raphanusinol is the monomer component with the highest content. The content of raphanusinol in the water extract of fried Semen Raphani is more than 8 times that of raw Semen Raphani, but raphanusinol can be completely lost if Semen Raphani is over-processed. Fried Semen Raphani can inhibit the activity of myrosinase in Semen Raphani and prevent the decomposition of raphanusinol.

[0014] Fried Fructus Perillae is slightly warm in nature, pungent in taste, and is attributed to the large intestine and lung meridians. It has the effects of reducing qi and resolving phlegm, relieving cough and asthma, moistening the intestines and promoting defecation, regulating qi and stomach, and relieving pain. It is clinically used for treating phlegm accumulation and qi stagnation, cough and asthma, dry intestines and constipation, etc., and can relieve the exterior and dispel cold, regulate qi and stomach.

[0015] Roasted Semen Armoraciae is pungent in taste and warm in nature, and it pertains to the lung and stomach meridians. The raw product is good at tonifying qi, relieving cough and asthma, warming middle energizer, dredging collaterals to stop pain, and moistening intestines to promote defecation, and it can be used to treat qi deficiency, cough and asthma, intestinal wind, constipation, etc. After being roasted, it moderates the pungent and dispersing nature of Semen Armoraciae, and it is good at regulating qi and resolving phlegm. It has the effects of warming lung to resolve phlegm, benefiting qi, resolving phlegm, and relieving pain, etc. It can be used to treat a series of diseases caused by cold stagnation.

[0016] Roasted Semen Galli is sweet in taste and neutral in nature, and it pertains to the spleen and stomach meridians. After being roasted, Semen Galli can correct odor and taste, and it is easier to take. Compared with the raw product, roasted Semen Galli has stronger effects of invigorating the spleen and eliminating accumulation. It is more effective in treating indigestion, food accumulation, spleen deficiency, diarrhea, and malnutrition in children. Modern pharmacological research shows that after being roasted, polysaccharides in Semen Galli are converted into monosaccharides, which are more easily dissolved. At the same time, the content of amino acids, which are the main chemical components and the main pharmacological and efficacy components of Semen Galli, also increases after being roasted.

[0017] Roasted Fructus Crataegi is sour and sweet in taste, and warm in nature. It pertains to the spleen, stomach, and liver meridians. It has the effects of digesting and invigorating the stomach, and promoting blood circulation to remove blood stasis. It is mainly used in the treatment of meat accumulation, stomach fullness, diarrhea, abdominal pain, and hernia pain. The main components of Fructus Crataegi are organic acids, phenolic acids, and triterpenoids. After being roasted, the content of organic acids increases, and the effects of digesting and invigorating the stomach, promoting blood circulation to remove blood stasis, resolving turbidity, and reducing fat also increase. It is mainly used in the treatment of meat accumulation, abdominal fullness, abdominal pain, diarrhea, blood stasis, postpartum blood stasis, chest pain, and hernia pain. Modern pharmacological research shows that roasted Fructus Crataegi has the effects of reducing blood lipids, reducing blood pressure, strengthening the heart, and preventing arrhythmia. Fructus Crataegi is also a good medicine for invigorating the spleen, opening the stomach, digesting and eliminating accumulation, and promoting blood circulation to remove blood stasis. Nie Chunxia et al. conducted a digestion-promoting experiment to detect the secretion of pepsin in food accumulation rat models, and the results showed that the intensity of regulating intestinal hormones was roasted Fructus Crataegi > roasted Fructus Crataegi > raw Fructus Crataegi. In addition, Liu Tianqi et al. studied the stimulating components limonene in each processed product, and the results showed that the content of limonene in roasted Fructus Crataegi was the lowest, which optimized the stimulation of the raw product on the gastrointestinal tract.

[0018] Pericarpium Citri Reticulatae is bitter and pungent in taste, and warm in nature. It pertains to the lung and spleen meridians. It has the effects of regulating qi and invigorating the spleen, and drying dampness and resolving phlegm. It is mainly used in the treatment of digestive and respiratory diseases. It is the most commonly used medicine for esophagus, stomach, and duodenum, and it can also be used to treat chest and hypochondriac pain, hernia, mammary nodules, mastitis, food accumulation, and abdominal pain. It mainly contains flavonoids, volatile oils, alkaloids, and polysaccharides. The main flavonoid is hesperidin, and the main volatile oil is limonene. Pharmacological research shows that it has the effects of anti-inflammatory, relieving asthma, inhibiting gastrointestinal smooth muscle movement, anti-tumor, anti-oxidation, and enhancing immunity.

[0019] Dandelion is bitter, sweet, cold. Liver, stomach. Has heat-clearing and detoxifying, swelling and knot-removing, diuretic and urinary stone-removing effects, and is mainly used for treating furuncle, mastitis, scrofula, red eye, sore throat, lung abscess, intestinal abscess, hot and humid sores, hot and painful urination, etc. Its chemical components are mainly flavonoids, phenolic acids, polysaccharides, etc., and it has anti-inflammatory, antioxidant, tumor inhibition, etc.

[0020] Glycyrrhiza is flat, sweet, and belongs to heart, lung, spleen and stomach. It has the effects of tonifying spleen and qi, clearing heat and detoxifying, eliminating phlegm and relieving cough, relieving pain and adjusting the effects of various drugs. It is used for spleen and stomach weakness, fatigue, shortness of breath, cough and phlegm, abdominal pain, and is used to relieve drug toxicity and violence. The main chemical components of glycyrrhiza are glycyrrhizinic acid, glycyrrhizin, glycyrrhizin, glycyrrhizin, glycyrrhizin, glycyrrhizin, etc. Modern research shows that glycyrrhizin has anti-inflammatory, adrenal cortex hormone-like effect, effect on myocardial ischemia, anti-cancer and anti-tumor effect; glycyrrhizin has anti-tumor, anti-free radical and anti-aging effects; glycyrrhizin has bacteriostatic effect, immune regulation effect, anti-virus effect and prevention and treatment of osteoarthritis; glycyrrhizin has anti-fibrosis effect and immune regulation effect; glycyrrhizin has anti-inflammatory, anti-malaria and anti-parasite, antibacterial effect; glycyrrhizin has anti-allergic reaction, anti-tumor, anti-inflammatory and immune regulation effect.

[0021] The postoperative ventilation recipe is composed of eight kinds of food-medicine homologous traditional Chinese medicines, including fried Semen Raphani, fried Semen Sinapis, fried Perilla Fruit, Taraxacum mongolicum, fried Endo-Testis, charred Haws, dried Tangerine Peel and licorice. The recipe contains hundreds of chemical components, and its mechanism is difficult to be explained due to its properties of multi-component, multi-target and multi-pathway. The three kinds of fried Semen Raphani, Semen Sinapis and Perilla Fruit are the monarch drugs in the recipe, which are also the components of the classical recipe Sanzi Yangqin Decoction. The recipe Sanzi Yangqin Decoction is from Han Shangyi's Medical Treatise, and has the effects of reducing qi and food, warming lung and resolving phlegm. In addition, the recipe can dredge meridians, disperse blood stasis and promote blood circulation, and plays an important role in treating blood stasis and obstruction in the intestinal tract. Fried Endo-Testis has the effects of clearing heat and resolving toxins, expelling phlegm and relieving cough, resisting inflammation and relieving pain, and promoting digestion, which can assist the monarch drug to promote the gastrointestinal motility. Charred Haws can reduce the stimulation to the gastrointestinal tract, and is good at promoting qi circulation, removing blood stasis, reducing food and stopping diarrhea. Dried Tangerine Peel, as a commonly used qi-regulating drug, has the effects of regulating qi and invigorating the spleen, which can promote the gastrointestinal motility and assist the monarch drug to regulate the gastrointestinal movement. Taraxacum mongolicum can promote the secretion of bile, regulate intestinal flora, reduce the expression of inflammatory factors, and increase the number of bifidobacterium and lactobacillus. Licorice, as a commonly used qi-tonifying drug, can not only tonify qi, expel phlegm, relieve cough, detoxify and relieve pain, but also harmonize the other drugs in the recipe. The whole recipe can promote the recovery of the gastrointestinal function after operation, promote the exhaust, facilitate the patients to eat and absorb nutrients from the gastrointestinal tract, and accelerate the recovery. The whole recipe can reduce qi, resolve phlegm, promote digestion and transportation, and promote the recovery of the gastrointestinal function after operation. The lung and the stomach and the intestines are treated together, the lung is ventilated and qi is reduced, the stomach is invigorated and transportation is assisted, the interior and the exterior are regulated together, the upper and the lower are treated together, and the gastrointestinal descending function is promoted to recover.

[0022] The fried Semen Raphani, fried Semen Sinapis and fried Perilla Fruit are processed from the concept of “the lung and the large intestine being interior-exteriorly related” in traditional Chinese medicine, and the nature of the three drugs is changed from ascending to descending after processing. The whole recipe can reduce the qi in the body, which is beneficial to the exhaust of the intestinal gas after operation. The recipe is prepared according to the best processing method of each drug, and is prepared into granules, which provides beneficial effects for the postoperative ventilation recipe.

[0023] The composition has significant curative effect and wide market application prospect through preclinical tests (animal experiments) and clinical tests. BRIEF DESCRIPTION OF DRAWINGS

[0024] Figure 1 is an HPLC chromatogram of the mixed control and the postoperative ventilation recipe;

[0025] Figure 2 is a total ion current chromatogram of UPLC-Q-TOF-MS / MS; wherein A: positive ion mode total ion current chromatogram; B: negative ion mode total ion current chromatogram;

[0026] Figure 3 is the effect of postoperative ventilation recipe on the ileum tissue pathological results of POI mice (HE staining, x200) A: sham operation group; B: model group; C: positive drug group; D: postoperative ventilation recipe group (24h); E: sham operation group; F: model group; G: positive drug group; H: postoperative ventilation recipe group (48h);

[0027] Figure 4 is the effect of postoperative ventilation recipe on the expression of TLR4, NF-κB p65 and MAPK p38 proteins in the ileum tissue of POI mice (400x);

[0028] Figure 5 is the relative expression of TLR4, NF-κB p65 and MAPK p38 proteins in the ileum tissue of mice in each group (compared with the sham operation group, ### P<0.001, ## P<0.01; compared with the model group, ***P<0.001, **P<0.01, *P<0.05);

[0029] Figure 6 is the intestinal microbial diversity analysis (n=5 / group) A: Wayne diagram; B: dilution curve; C: rank abundance curve; D: PCoA analysis of β diversity analysis; E: NMDS analysis of β diversity analysis;

[0030] Figure 7 is the intestinal flora Alpha diversity analysis (A, B represent ACE, Chao1 index respectively) data analyzed by one-way ANOVA (mean ± standard deviation, n=5 / group) compared with the sham operation group, # P<0.05, ## P<0.01, ### P<0.001; compared with the model group, *P<0.05, **P<0.01, ***P<0.001 vs POI group;

[0031] Figure 8 is the species structure diagram at different levels in the intestinal microbial community (n=5 / group) A: door level; B: class level; C: genus level; D: species level (TQ represents the 24h postoperative ventilation recipe group);

[0032] Figure 9 is the LEfSe analysis (n=5 / group);

[0033] Figure 10 is the carbon powder propulsion rate (%) of rat small intestine * P<0.05, ** P<0.01, *** P<0.001, **** P<0.001);

[0034] Figure 11 is the spleen index of rats * P<0.05, **P<0.01, *** P<0.001, **** P<0.001);

[0035] Figure 12 is a rat small intestine tissue HE staining diagram (left 10x, right 40x);

[0036] Figure 13 is the effect of postoperative ventilation side on the serum IL-6, TNF-α, CRP levels of postoperative ileus rats, and the IL-6, TNF-α, MPO levels of small intestine tissue * P<0.05, ** P<0.01, *** P<0.001, **** P<0.001);

[0037] Figure 14 is the change of ZO-1 expression in the small intestine tissue of 3-5 POI rats;

[0038] Figure 15 is the serum metabolomics of rats (A-D: OPLS-DA analysis diagram under positive ion mode, E-H: OPLS-DA analysis diagram under negative ion mode);

[0039] Figure 16 is a rat serum differential metabolite clustering heat map (C: control group, M: model group, G: postoperative ventilation side high dose group);

[0040] Figure 17 is a rat serum endogenous differential metabolite KEGG pathway enrichment diagram. DETAILED DESCRIPTION

[0041] The following examples will further illustrate the present application, which are only used to illustrate the present application without any limitation on the present application.

[0042] Example 1

[0043] 1. Granule preparation

[0044] 1.1. Extraction

[0045] 1.1.1. Determination of content determination method

[0046] (1) Chromatographic conditions and system suitability test

[0047] Take octadecylsilane bonded silica gel as the filler, take acetonitrile as the mobile phase A, take 0.1% phosphoric acid as the mobile phase B, perform gradient elution according to the following table (Table 1), and the detection wavelength is 326nm, 283nm.

[0048] Table 1 Mobile phase proportion table

[0049] (2) Preparation of mixed control solution

[0050] Take mustard alkali thiocyanate control appropriate, precision, add methanol solution to make every 1 mL containing mustard alkali thiocyanate 196.00 μg; Take rosemary acid control appropriate, precision, add methanol solution to make every 1 mL containing rosemary acid 80.26 μg; Take chicoric acid control appropriate, precision, add methanol solution to make every 1 mL containing chicoric acid 169.83 μg; Take hesperidin control appropriate, precision, add methanol solution to make every 1 mL containing hesperidin 423.36 μg.

[0051] Respectively 0.5 mL mustard alkali thiocyanate control solution, 1 mL rosemary acid control solution, 1 mL chicoric acid control solution, 1 mL hesperidin control solution to 10 mL volumetric flask, add methanol dilution to the calibration line.

[0052] (3) Preparation of test solution

[0053] Take compound decoction liquid 2 mL to 10 mL volumetric flask, add methanol dilution to the calibration line, shake, filter, take the filter liquid, that is.

[0054] (4) Determination method

[0055] Respectively 10 μL of control solution and test solution, inject liquid chromatograph, determination, that is.

[0056] 1.1.2. Single factor investigation

[0057] This study according to the "medical institutions of traditional Chinese medicine decoction room management specification" in the provisions and requirements, determine the soaking time, extraction time, water, extraction times as the influencing factors of extraction process conditions, and screening, this experiment adopts the multi index comprehensive score, based on the theory of traditional Chinese medicine compatibility, the pharmacodynamic action of each effective component, combined with the analytic hierarchy process to analyze the index weight coefficient.

[0058] Comparison of mustard alkali thiocyanate, rosemary acid, chicoric acid, hesperidin, the relative importance of 5 evaluation indexes of extraction rate, and constitute pairwise comparison matrix, see table 2. According to the results of the table, the data is normalized, the weight coefficient of mustard alkali thiocyanate, rosemary acid, chicoric acid, hesperidin and extraction rate is 0.1944, 0.1944, 0.0930, 0.0930, 0.4251, respectively, the consistency ratio factor CR=0.0022<0.1, indicating that the judgment matrix meets the consistency requirements, and the weight coefficient is effective.

[0059] Extraction rate is: extraction rate (%) = [extraction weight (g) × sample total volume (mL)] / [herbal medicine weight (g) × sample amount (mL)] × 100%;

[0060] The overall score is: Y = (0.1944 × sinigrin thiocyanate content / maximum sinigrin thiocyanate content + 0.1944 × rosmarinic acid content / maximum rosmarinic acid content + 0.0930 × chicoric acid content / maximum chicoric acid content + 0.0930 × hesperidin content / maximum hesperidin content + 0.4251 × yield value / maximum yield value) × 100.

[0061] Table 2 Judgment Matrix for Comparison of Index Components

[0062] (1) Selection of soaking time

[0063] Weigh out four portions of the prescribed amount of medicinal slices, add 8 times the amount of water and soak for 20, 35, 50 and 65 minutes, decoct for 30 minutes and filter; add 6 times the amount of water to the medicinal slices for the second time, decoct for 30 minutes and filter, combine the filtrates, take samples and determine according to section 4.3.2, the comprehensive scores are 90.43, 95.19, 93.89 and 95.42.

[0064] Experimental results showed that soaking times of 35 min and 65 min yielded high and similar overall scores. To improve efficiency, the soaking time was ultimately determined to be 35 min.

[0065] (2) Selection of water volume

[0066] Weigh out four portions of the prescribed amount of medicinal slices, add 6, 8, 10, and 12 times the amount of water respectively, soak for 35 minutes, decoct for 30 minutes, and filter. Add 6 times the amount of water to the medicinal slices for the second time, decoct for 30 minutes, filter, combine the filtrates, take samples, and determine according to section 4.3.2. The comprehensive scores are 85.34, 85.83, 93.84, and 97.18.

[0067] Experimental results show that the optimal water addition ratios are 8, 10, and 12.

[0068] (3) Selection of extraction time

[0069] Weigh out four portions of the prescribed amount of medicinal slices, soak them in 8 times the amount of water for 35 minutes, and decoct them for 20, 40, 60 and 80 minutes respectively, and filter them. For the second decoction, add 6 times the amount of water to the medicinal slices, decoct for 30 minutes, filter them, combine the filtrates, take samples, and determine them according to section 4.3.2. The comprehensive scores are 96.73, 97.46, 97.32 and 92.37.

[0070] Experimental results show that the optimal simmering times are 20, 40, and 60 minutes.

[0071] (4) Selection of extraction times

[0072] Weigh out four portions of the prescribed amount of medicinal slices and number them ①, ②, ③, and ④.

[0073] ① The prescription amount of decoction pieces was added with 8 times water to soak for 35 min, decocted for 30 min, filtered to sample, and determined according to 4.3.2.

[0074] ② The prescription amount of decoction pieces was added with 8 times water to soak for 35 min, decocted for 30 min, filtered; the residue was added with 6 times water, decocted for 30 min, filtered, the filtrates were combined, sampled and determined according to 4.3.2.

[0075] ③ The prescription amount of decoction pieces was added with 8 times water to soak for 35 min, decocted for 30 min, filtered; the residue was added with 6 times water, decocted for 30 min, filtered; the residue was added with 6 times water, decocted for 30 min, filtered, the filtrates were combined, sampled and determined according to 4.3.2.

[0076] ④ The prescription amount of decoction pieces was added with 8 times water to soak for 35 min, decocted for 30 min, filtered; the residue was added with 6 times water, decocted for 30 min, filtered; the residue was added with 6 times water, decocted for 30 min, filtered; the residue was added with 6 times water, decocted for 30 min, filtered, the filtrates were combined, sampled and determined according to 4.3.2.

[0077] The comprehensive scores were 48.04, 78.58, 92.47 and 100.00.

[0078] The experimental results showed that the scores of decocting for three times and four times were higher, and decocting for one, two and three times were selected for improving the decocting efficiency.

[0079] 1.1.3. Orthogonal experiment

[0080] According to the single factor investigation experimental results, the water addition amount (A), extraction time (B) and extraction times (C) were taken as the investigation factors, and L9 (34) orthogonal experiment was arranged. The factor level table was shown in Table 3. The investigation indexes were sinapine thiocyanate, rosmarinic acid, chicoric acid, hesperidin and the extract yield, the experimental arrangement and results were shown in Table 4, the direct analysis results were shown in Table 5, and the variance analysis was shown in Table 6. 4

[0081] Table 3 Factor level table

[0082] Table 4 Orthogonal experiment design and results of extraction process

[0083] Table 5 Direct analysis results

[0084] Table 6 Variance analysis

[0085] ​The results of the test show that the comprehensive score results of Table 4 show that the influence of the active ingredient extraction results of the postoperative ventilation prescription is in the order of C > B > D > A; the variance analysis results of Table 6 show that factor B has a significant influence on the extraction process. The range analysis of the comprehensive score shows that A3 is the best decoction time, but for sinapine thiocyanate and chicoric acid, A2 decoction time is more appropriate, and the content of sinapine thiocyanate and chicoric acid is larger than that of other active ingredients. Finally, A2B3C3 is selected as the best water extraction process.

[0086] 1.1.4. Optimization of the number of decoctions

[0087] According to the extraction conditions selected by the orthogonal test, the total decoction water decoction liquid and the third decoction water decoction liquid were sampled, respectively, and were determined according to the method in 4.3.2, and the total contribution rate of the third decoction liquid was calculated. Details are shown in Table 7.

[0088] Table 7 Comparison of the third decoction water extract and the decoction water extract

[0089] The test results show that the contribution rate of the third decoction water extract to the total decoction water extract is 12.39%, which is less than 15%. Considering the production cost, the number of decoctions is finally determined to be 2 times. That is, 12 times the amount of water is added, soaked for 35 min, decocted for 40 min, and filtered; 6 times the amount of water is added to the drinking pieces for the second time, decocted for 30 min, and filtered, and the above filtrates are combined.

[0090] 1.2. Concentration

[0091] After extraction, the concentrated liquid is passed through a 100-mesh screen.

[0092] 1.3. Granulation

[0093] An appropriate amount of dextrin is placed in a fluidized bed, the inlet air temperature is set to 100°C, and when the material temperature rises to 70°C, the liquid feeding starts, the liquid feeding speed is controlled at 80-150 r / min, the atomizing pressure is 0.2 MPa outside and 0.15 MPa inside, the spraying is stopped, and the drying continues at 70°C to obtain granules. Since granulation may result in powders without granulation or granules with excessive adhesion leading to caking, the granules obtained need to be sieved. The granules obtained are sieved through 20-mesh and 80-mesh vibrating screens, and the granules between 20-80 mesh are the target granules. The HPLC chromatogram of the postoperative ventilation prescription is shown in Figure 1.

[0094] Example 2

[0095] 1. First preparation method

[0096] 1.1. Granule preparation

[0097] The pharmaceutical composition is composed of:

[0098] Table 8

[0099] 1.1.1. Extraction

[0100] Preferably, 3.0 kg of fried Raphani Semen, 1.0 kg of fried Perillae Fructus, 1.0 kg of fried Semen Ranunculi Ternati, 3.0 kg of Herba Taraxaci, 1.0 kg of charred Fructus Crataegi, 0.9 kg of fried Semen Galli, 0.6 kg of Pericarpium Citri Reticulatae, and 0.3 kg of Glycyrrhizae Radix et Rhizoma are weighed in proportion, and 12 times of water (12 times of the single-pot point of the total amount of the ingredients) is added. After being soaked for 35 min, the mixture is heated to boiling, and kept at a slight boil for 0.75 h. Then, 6 times of water (6 times of the single-pot point of the total amount of the ingredients) is added, the mixture is heated to boiling, and kept at a slight boil for 0.5 h. The decoction is combined, filtered, and then fluidized bed granulation is performed by adding an appropriate amount of malt dextrin.

[0101] 1.1.2. Concentration

[0102] After the extraction is completed, the concentration process is performed. The concentrated liquid is passed through a 100-mesh screen, and the weight is 10.49 kg.

[0103] 1.1.3. Granulation

[0104] An appropriate amount of dextrin is placed in the fluidized bed, and the inlet air temperature is set to 100°C. When the material temperature rises to 70°C, the liquid feeding is started, and the feeding speed is controlled at 80-150 r / min. The spraying is ended at an external 0.2 MPa and an internal 0.15 MPa. Then, the drying is continued at 70°C to obtain granules. Since the granulation may result in powders without granulation or granules with excessive adhesion leading to caking, the obtained granules need to be sieved. The obtained granules are sieved through 20-mesh and 80-mesh vibrating screens, respectively. The granules between 20-mesh and 80-mesh are the target granules.

[0105] 2. Preparation of postoperative ventilation medicinal composition decoction pieces

[0106] Fried Raphani Semen is prepared as follows: Raphani Semen is the dried mature seed of Raphanus sativus L. of the Brassicaceae family. An appropriate amount of Raphani Semen decoction pieces is placed in a frying machine, and the frying temperature is set to 220°C. The rotation speed of the frying machine is set to 30 r / min, and the frying time is set to 5 min. After being crushed, the fried Raphani Semen is ready for use. The fried Raphani Semen prepared according to the preparation condition has a red-brown surface, is easy to crush, and has a yellow interior.

[0107] Raphanus sativus L. has the effects of eliminating food retention and reducing flatulence, and is used for treating food retention, abdominal pain, constipation, dysentery, and phlegm accumulation. Raphanus sativus L. is a typical Chinese medicine for “different treatment for raw and cooked, ascending for raw and descending for cooked”, and the roasted Raphanus sativus L. has the effect of eliminating food retention and reducing flatulence, and is commonly used in clinic. Modern studies have shown that Raphanus sativus L. mainly contains glucosinolates, sulfur derivatives, alkaloids, flavonoids, fatty acids, volatile oils, and polysaccharides. Glucosinolates are the characteristic components before and after processing, and raphanus sativus L. is the monomer component with the highest content in glucosinolates. The content of raphanus sativus L. in water extract of roasted Raphanus sativus L. is increased by more than 8 times compared with raw product, but if over-processed, raphanus sativus L. can be lost completely. Roasted Raphanus sativus L. can inhibit the activity of myrosinase in Raphanus sativus L. and prevent the decomposition of raphanus sativus L.

[0108] Preparation of roasted Perilla frutescens: Perilla frutescens is the dried mature fruit of Perilla frutescens (L.) Britt. of Lamiaceae. Take the raw product of Perilla frutescens decoction pieces, place them in a frying machine, set the frying temperature to 220℃, the rotating speed of the frying machine to 30r / min, and the frying time to 3min, crush them, and reserve for use. According to the preparation conditions, the surface of the prepared roasted Perilla frutescens decoction pieces is grayish brown, with fine cracks, and a burnt aroma.

[0109] Preparation of roasted Semen Brassicae: Semen Brassicae is the dried mature seed of Brassica juncea (L.) Czern.et Coss. of Brassicaceae. Take the raw product of Semen Brassicae, place it in a frying machine, set the frying temperature to 220℃, the rotating speed of the frying machine to 30r / min, and the frying time to 3min, crush it, and reserve for use. According to the preparation conditions, the surface of the prepared roasted Semen Brassicae decoction pieces is dark yellow to brownish, with occasional burnt spots, and a spicy aroma.

[0110] Preparation of charred Fructus Crataegi: Fructus Crataegi is the dried mature fruit of Crataegus pinnatifida Bge. Take the raw product of Fructus Crataegi, place it in a frying machine, set the frying temperature to 300℃, the rotating speed of the frying machine to 30r / min, and the frying time to 15min. According to the preparation conditions, the surface of the prepared charred Fructus Crataegi decoction pieces is dark brown, and the inside is yellowish brown, with a burnt aroma.

[0111] Preparation of roasted Gigeriae Cibi: Gigeriae Cibi is the dried inner wall of the gizzard of Gallus gallus domesticus Brisson. Take the raw product of roasted Gigeriae Cibi, place it in a frying machine, set the frying temperature to 230℃, the rotating speed of the frying machine to 30r / min, and the frying time to 2min. According to the preparation conditions, the surface of the prepared roasted Gigeriae Cibi decoction pieces is dark yellowish brown or burnt yellow, and appears granular or micro-bubble under a magnifying glass. It is brittle and has a shiny fracture surface.

[0112] 3. Detection of postoperative ventilation medicinal composition decoction pieces

[0113] The first part is based on UPLC-QTOF-MS / MS analysis of the whole composition of postoperative ventilation prescription

[0114] 1.1 Experimental method

[0115] 1.2 Preparation of test solution

[0116] Weigh about 1.5g of postoperative ventilation prescription granules, grind in a mortar, and accurately weigh 1g. Add 5mL of 75% methanol solution, weigh the mass, shake well, and extract for 30min. Cool, re-weigh the mass, and make up the lost mass with 75% methanol. Centrifuge at 12000r·min-1 for 10min, pass the supernatant through a 0.22μm microporous filter membrane, and take the filtrate to obtain the solution.

[0117] 1.3 Chromatographic conditions

[0118] Chromatographic column: ACQUITY HSS T3 chromatographic column (2.1×100mm, 1.8μm). Mobile phase: 0.05% formic acid water (A) and acetonitrile (B). Column temperature 35℃, injection volume 2μL. Gradient elution program as shown in Table 9:

[0119] Table 9 Chromatographic gradient table

[0120] 1.4 Mass spectrometry conditions

[0121] AB SCIEX TripleTOF TM 5600 liquid chromatograph-mass spectrometer was used for detection, with electrospray ionization (ESI) source and positive and negative ion modes. Ionization temperature (TEM): 550.0℃; atomization gas (GS1): 55psi; auxiliary heating gas (GS2): 55psi; gas curtain gas (CUR): 30psi. The first mass spectrometry acquisition range was m / z 50-1500, with cumulative time 0.15s; DP: 60V; CE: 45eV. The second mass spectrometry was collected in IDA mode, with acquisition range m / z 50-1500, cumulative time 0.1s, DP: 60V; CE: 45eV; CES: 20; IRD 67; IRW 25. Molecular weight error 50mDa. The relative molecular mass accuracy was automatically calibrated using the tuning liquid delivery system (CDS) of AB company.

[0122] 2. Experimental results

[0123] By local database combined with reference and literature comparison analysis, 130 components were identified and inferred from the postoperative ventilation formula granules, and the total ion chromatogram is shown in Figure 2. Among them, 62 are flavonoids, 9 are saponins, 10 are terpenes, 1 is an alkaloid, 3 are sugars, 4 are phenylpropanoids, 6 are amino acids, 24 are organic acids, and 11 are other components.

[0124] 4. Preliminary evaluation of granule pharmacodynamics - mouse experiment

[0125] 1 Experimental method

[0126] Model group (intestinal manipulation group):

[0127] The classic intestinal manipulation (IM) was used to manufacture the mouse postoperative ileus (POI) model. First, 4% chloral hydrate was injected intraperitoneally for anesthesia, with a dose of 0.1 mL / 10g. After anesthesia, the mouse was placed on the operating table in a supine position. The abdomen was routinely shaved and disinfected, and sterile gauze was laid on the incision. A 2cm incision was made in the midline of the abdomen. A wet cotton swab soaked in normal saline was used to wipe the small intestine repeatedly, 4 minutes each time, for 3 times. The small intestine was returned in turn, and the abdominal muscle layer and skin layer were sutured with 5-0 absorbable suture. The abdominal wound was disinfected again to prevent infection. The mouse was placed under warm light for recovery.

[0128] Sham operation group (open abdomen without disturbing the small intestine group):

[0129] This group did not disturb the small intestine after laparotomy, and the rest of the operation was the same as above.

[0130] Observation group (intestinal manipulation + traditional Chinese medicine group in this application):

[0131] Eight cases of postoperative ileus mice were treated with postoperative ventilation formula granules for 24h observation, and another 8 cases of postoperative ileus mice were treated with postoperative ventilation formula granules for 48h observation. The observation indexes were the expression levels of serum inflammatory factors (IL-6, IL-10, TNF-α) and brain-gut peptides (MTL, SS) in mice, hematoxylin-eosin (HE) staining to observe the morphological changes of ileum tissue, immunohistochemical method (IHC) to observe the expression of TLR4, NF-κB p65 and MAPK p38 proteins in mouse ileum tissue, and 16s rRNA sequencing method to analyze the changes of intestinal flora in the colon contents of mice in each group.

[0132] Positive western medicine group (intestinal manipulation + mosapride citrate group):

[0133] Drug intervention was mosapride citrate gavage, and the rest was the same as above

[0134] Results: Postoperative ventilation treatment can treat postoperative intestinal injury by reducing systemic inflammatory response in mice, restoring brain-gut peptide hormone levels, and improving intestinal flora disorders, and has good efficacy 24 hours after administration. The grouping is shown in the table below.

[0135] Table 10

[0136] According to the body surface area conversion method and the clinical dose of each drug, the drug dosage of mice was calculated as follows: mosapride citrate 1.95 mg / (kg·d), postoperative ventilation prescription 1.88 g / (kg·d), and the initial intragastric administration was performed at 6 hours after the operation.

[0137] The data were statistically analyzed and statistical graphs were drawn using Spss21.0h and Graphpad Prism9.0 software, and the data were expressed as mean ± standard deviation Statistical analysis: One-way ANOVA was used for comparison between multiple groups, and P<0.05 was considered statistically significant.

[0138] 2 Experimental results

[0139] 2.1 Effect of postoperative ventilation prescription on the histopathological results of ileum tissue in POI mice

[0140] As shown in Figure 3, compared with the sham operation group, the ileum tissue of POI mice showed intestinal wall atrophy, small intestinal villus axis edema and congestion, villus atrophy and shortening, reduced number, mucosal capillary congestion, and different degrees of inflammatory cell infiltration in the mucosal surface and muscle layer. However, these pathological changes were alleviated to different degrees after intervention with positive western medicine (mosapride citrate) and postoperative ventilation prescription.

[0141] 2.2 Effect of postoperative ventilation prescription on serum inflammatory factors and brain-gut peptide levels in POI mice

[0142] As shown in Tables 11 and 12, compared with the sham operation group, the contents of pro-inflammatory cytokines TNF-α and IL-6 in the POI mice of the model group were significantly increased, and the content of anti-inflammatory factor IL-10 was significantly decreased. In addition, the content of MTL in the serum of the POI group was significantly decreased, while the content of SS was significantly increased. Both positive western medicine (mosapride citrate) and postoperative ventilation prescription treatment can reverse the changes of inflammatory factors and brain-gut peptides. The results show that postoperative ventilation prescription can not only down-regulate the expression of pro-inflammatory factors and up-regulate the expression of anti-inflammatory factors, but also improve the disorder of brain-gut peptide hormones.

[0143] Table 11 Changes of inflammatory factors and gastrointestinal hormones in 24h POI mice

[0144] Table 12 Changes of inflammatory factors and gastrointestinal hormones in 48h POI mice in vivo

[0145] a: P < 0.05 compared with the sham operation group; b: P < 0.05 compared with the model group.

[0146] 2.3 Immunohistochemical detection of changes in TLR4, NF-κB p65 and MAPK p38 protein expression in ileum tissues of POI mice

[0147] As shown in Figures 4 and 5, compared with the sham operation group, the expression levels of TLR4, NF-κB p65 and MAPK p38 in the POI model group were significantly increased, and the expression levels of each protein were significantly reduced after administration of the positive drug and the postoperative ventilation granules, indicating that the postoperative ventilation granules can reduce the expression of inflammatory proteins in the TLR4, NF-κB and MAPK pathways in ileum tissues.

[0148] 2.4 Microbial analysis of intestinal flora in colon contents of mice

[0149] Table 13 Statistical analysis of Alpha diversity index

[0150] To reveal the effects of postoperative ventilation formula on the richness, evenness, uniformity and diversity of intestinal microbial community of mice, we performed Alpha diversity analysis on the Venn diagram, dilution curve and rank abundance curve of the samples. The Venn diagram illustrates the number of common and unique species characteristics between different groups, and there are 334 operational taxonomic units (OTUs) shared by all groups. There are more OTUs between the TQ group (postoperative ventilation formula treatment group) and the Sham group (sham operation group) (OUTs = 551) than between the POI group (postoperative ileus model group) and the Sham group (OUTs = 380). The dilution curve shows that the sequencing depth has basically covered all species in the sample. The species richness and evenness in the sample can be reflected on the rank abundance curve. Compared with the POI group, the horizontal axis curve range of the Sham group and the TQ group is larger and the curve is smoother, indicating that the species richness and evenness of the Sham group (sham operation group) and the TQ group (postoperative ventilation formula group) are higher. The Alpha diversity index analysis is shown in Table 13. The Coverage value of all samples is close to 0.99, indicating that the sequencing result is sufficient. The Chao1, Simpson, Shannon and PD indices of the POI group are significantly lower than those of the control group, and the TQ group is closer to the control group. From the data, TQ can increase the richness of intestinal flora. Beta diversity analysis, including Principal coordinates analysis (PCoA) and Non-Metric Multi-Dimensional Scaling (NMDS), is used to compare the similarity of species diversity between different populations. The results show that the TQ group is closer to the control group than the POI group, which means that TQ can restore the disorder of intestinal flora after administration. See Figures 6 and 7.

[0151] The species data of each group of mice at the door, class, genus and species level were analyzed, and the proportion of the top 10 species at different classification levels was compared. At the door level (Figure 8A), each group was dominated by Firmicutes, Bacteroidetes, Proteobacteria and Verrucomicrobia. The number of Bacteroidetes in POI mice decreased significantly, the number of Proteobacteria increased significantly, and these changes could be reversed after TQ administration. At the class level (Figure 8B), each group was dominated by Bacteroidia, Bacilli, Clostridia and Gammaproteobacteria. The relative abundance of Bacteroidia and Clostridia in the model group decreased compared with the sham operation group, and the relative abundance of Bacilli and Gammaproteobacteria increased compared with the sham operation group. TQ administration alleviated these changes to some extent. At the genus level (Figure 8C), compared with the sham operation group, the model group had increased levels of Ligilactobacillus and Escherichia_Shigella, and decreased levels of Muribaculaceae and Lachnospiraceae. After TQ administration, the above bacterial species levels were adjusted to different degrees (P<0.05). Similarly, changes in intestinal flora at the genus and species levels could be clearly observed (Figures 8C-D), and TQ treatment could restore the disturbed flora to the normal baseline, as shown in Figure 8. LEfSe (Linear discriminant analysis Effect Size) was used to find specific flora with statistical differences between different groups. Compared with the sham operation group, the relative proportion and abundance of Shigella (P<0.05), Enterobacteriaceae (P<0.05) and Enterobacteriaceae (P<0.05) in the intestinal flora of the POI group increased. After TQ gavage, the beneficial bacteria NK4A214 (P<0.05) increased, as shown in Figure 9.

[0152] 5. Animal test - rat experiment

[0153] Based on the previous experimental basis, a positive Chinese medicine control group was added, and the postoperative ventilation decoction gavage time was determined to be one day.

[0154] (1) Experimental grouping

[0155] 60 rats were randomly divided into blank group, model group, postoperative ventilation grouping high / low dose, positive western medicine group (Mosapride Citrate), positive traditional Chinese medicine group (Si Mo Decoction), 10 rats in each group. Mosapride citrate group 1.35 mg / kg (6.3 times the human dose), Si Mo Decoction group 5.4 mg / kg (6.3 times the human dose), postoperative ventilation prescription low dose group 1.305 g / kg (6.3 times the human dose), high dose group 2.61 g / kg (12.6 times the human dose).

[0156] The specific grouping is shown in the following table:

[0157] Table 14 Rat experimental grouping

[0158] (2) Experimental method

[0159] 2.1 Sample collection

[0160] One day before the last administration, the rats in each group were fasted but not watered. 6 hours after the last administration, the rats in each group were ip 10% chloral hydrate (10%) anesthesia, the dose was 0.3 mL / 100 g, and the abdominal cavity was opened in a sterile environment. The physiological condition of the rat intestine was observed and recorded. Blood was taken from the posterior abdominal aorta, and after standing for 40 min, it was centrifuged at 3500 r / min for 10 min. The supernatant was centrifuged again at 14000 r / min for 10 min at 4℃, and the supernatant was collected and stored at -80℃ for standby. Rapid separation of multiple segments of rat small intestine, 3-5 cm per segment, gently washed with normal saline and placed on sterile filter paper to absorb part of the water, finally put into 4% paraformaldehyde for fixation.

[0161] 2.2 Small intestine charcoal propelling rate

[0162] 6 hours after the last administration, 1 mL of charcoal liquid (3 g of charcoal was dissolved in 50 mL of 0.5% sodium carboxymethyl cellulose solution) was given to each rat. 25 min later, anesthesia was performed, blood was taken from the abdominal aorta, and the whole small intestine was removed. The length of the small intestine was measured, and the length of the charcoal propelling from the pylorus to the front of the charcoal was also measured. The charcoal propelling rate was calculated according to the formula:

[0163] Charcoal propelling rate (%) = ink propelling length (cm) / total length of small intestine (cm) x 100%;

[0164] 2.3 Spleen / body weight index

[0165] The rat spleen was taken out, quickly placed on filter paper to absorb water, weighed, and the spleen index was calculated to two decimal places.

[0166] Spleen index = spleen weight (mg) / body weight (g)

[0167] 2.4 Immunofluorescence detection of ZO-1 expression in rat small intestine tissue

[0168] About 3 cm of small intestine tissue was taken from the end of the intestine after the rats were sacrificed, fixed in 4% paraformaldehyde for 24 h, and then paraffin-embedded. The paraffin section was deparaffinized to water, and then sequentially placed in dimethylbenzene I for 15 min, dimethylbenzene II for 15 min, anhydrous ethanol for 5 min, 85% ethanol for 5 min, 75% ethanol for 5 min, ddH2O for 5 min, and PBS for 5 min.

[0169] After deparaffinization to water, the tissue section was placed in an antigen repair solution and heated in a microwave oven for antigen repair: medium fire for 8 min, stop fire for 58 min, and low fire for 7 min. During this process, the buffer should be prevented from excessive evaporation, and the slice should not be dried. After natural cooling, the slide was placed in PBS (pH 7.4) and shaken for 3 times for 5 min each time on a decolorizing shaker.

[0170] Circle serum blocking: after the slice was gently shaken and dried, a histological pen was used to draw a circle around the tissue, and 5% BSA blocking solution was dropped into the circle for incubation for 30 min.

[0171] Add ZO1 primary antibody: after the blocking was completed, the blocking solution was discarded, and the primary antibody diluent was prepared according to the primary antibody instructions using 1% BSA. The prepared primary antibody diluent was dropped into the circle, covered the tissue, and the slice was placed flat in a wet box for incubation overnight at 4°C.

[0172] Add secondary antibody: the primary antibody diluent was discarded, the slide was placed in PBS and shaken for 3 times for 5 min each time on a decolorizing shaker in the dark, the PBS was discarded, the slice was slightly shaken and dried, and the fluorescent secondary antibody diluent diluted with 1% BSA of the same species as the primary antibody was added in the circle, covered the tissue, and incubated at room temperature in the dark for 50 min.

[0173] DAPI restain the cell nucleus: the slide was placed in PBS and shaken for 3 times for 5 min each time on a decolorizing shaker. After the slice was slightly shaken and dried, DAPI staining solution was added in the circle, and incubated at room temperature in the dark for 10 min.

[0174] Quench the tissue autofluorescence: the slide was placed in PBS (pH 7.4) and shaken for 3 times for 5 min each time on a decolorizing shaker. Autofluorescence quencher was added in the circle for 5 min, and then washed with running water for 10 min. After the slice was slightly shaken and dried, 25 μL of anti-fluorescence quenching mounting agent was added for mounting. Microscopy and photography: the slice was observed under a fluorescence microscope and the image was collected.

[0175] 7. Rat serum metabolomics experiment

[0176] The blood was taken from the abdominal aorta of the rats 40 min after the last administration, centrifuged at 3500 r / min for 10 min, and the supernatant was centrifuged again at 14000 r / min for 10 min at 4°C. The supernatant was collected, divided, and stored in a refrigerator at -80°C for standby.

[0177] Each serum sample was slowly thawed at 250 μL. After thawing, 50 ul of each serum sample was taken and mixed together as a quality control sample (QC). Two volumes of pre-cooled mass spectrometry grade methanol + mass spectrometry grade acetonitrile (1:1 by volume) were added to each serum sample and mixed QC sample, vortexed for 2-5 min to fully mix the protein precipitate with the serum sample, then placed in a 4°C centrifuge for natural sedimentation for 30 min, and then centrifuged at 4°C at 12000 r / min -1 for 15 min, and the supernatant was evaporated to dryness with nitrogen. The dried sample was reconstituted with 100 μL of mass spectrometry grade acetonitrile, vortexed for 1 min, and then centrifuged at 4°C at 12000 r / min -1 for 15 min, and the supernatant was injected.

[0178] ACQUITY HSS T3 column (100 mm x 2.1 mm, 1.8 μm); mobile phase 0.05% formic acid aqueous solution (A) - acetonitrile (B); flow rate 0.3 m L / min -1 , column temperature 40°C, injection volume 2 μL; gradient as shown in the following table.

[0179] Table 15 Chromatographic gradient table

[0180] The data acquisition was selected in full scan automatic mode, electrospray ion source (ESI), positive and negative ion mode scanning, scan range m / z 50-1000, mass spectrometry conditions as shown in the following table.

[0181] Table 16 Mass spectrometry conditions

[0182] 2.5 Experimental results

[0183] 2.5.1 Rat small intestine charcoal propelling rate

[0184] The results of the rat small intestine propelling rate are shown in Table 17 and Figure 10, and * represents that the experimental results are statistically significant compared with the model group. The results show that after the POI rats take the drug, the small intestine charcoal propelling rate increases significantly (P<0.05), and shows a dose-dependent manner.

[0185] Table 17 Results of rat small intestine charcoal propelling experiment

[0186] 2.5.2 Rat spleen-weight index

[0187] Table 18 Results of rat small intestine charcoal propelling experiment

[0188] Note: * indicates a significant difference (P < 0.05) compared with the model group

[0189] Reference is made to Fig. 11.

[0190] 2.5.3 Rat small intestine tissue HE staining

[0191] After the small intestine tissue section is deparaffinated, hydrated, etc., the section is dyed using an HE kit, and after completion, the glass slide is sealed with neutral glue, and an optical microscope is used for observation and photography.

[0192] HE staining is one of the reliable means for observing histopathology. In this study, the general morphological structure of the normal control group and the small intestine tissue after small intestine interference was observed by HE staining technology to explore the protective effect of postoperative ventilation recipe on the small intestine barrier. The HE staining results of the small intestine tissue sections of the rats in each group are shown in Fig. 12.

[0193] As can be seen from Fig. 12, the pathological performance of the small intestine tissue of the control group rats is normal, the tissue structure is complete, the epithelial cells are arranged closely, and no obvious damage is found in the mucosa layer. In comparison, the small intestine tissue cells of the model group rats undergo necrosis and apoptosis, are arranged in disorder, and the intercellular tight junction structure is destroyed. After drug intervention, the degree of damage to the small intestine epithelial structure gradually decreases, among which the positive western medicine group and the postoperative ventilation recipe high dose group have the best improvement effect. Compared with the model group, the small intestine cells in the positive western medicine group and the postoperative ventilation recipe high dose group are normal in shape, arranged in order, the epithelial structure is complete, and the boundary is clear. This indicates that the postoperative ventilation recipe can treat the damaged small intestine barrier and restore the integrity of the small intestine epithelial cell structure.

[0194] 2.5.4 Determination of indicators in rat small intestine tissue and serum

[0195] The determination of TNF-α, IL-6, CRP in serum and TNF-α, IL-6, MPO in tissue is strictly in accordance with the kit instructions, and reference is made to Fig. 13.

[0196] 2.5.5 ZO-1 expression in intestinal tissue of rats in each group

[0197] The ZO-1 expression in the intestinal tissue of rats in each group is shown in Fig. 14. As can be seen from the figure, compared with the control group, the ZO-1 expression in the intestinal tissue of the POI model group rats is significantly reduced, and the ZO-1 expression has different degrees of recovery after administration, indicating that the postoperative ventilation recipe has a repairing effect on the intestinal barrier.

[0198] 2.5.6 Serum metabolomics of rats

[0199] The UPLC-Q-TOF-MS / MS technology was used to collect data of serum of rats in different groups in positive and negative ion modes. The obtained data was introduced into SIMCA 14.1 for orthogonal partial least squares discriminant analysis (OPLS-DA) and principal component analysis (PCA), as shown in the following figure. The QC samples were well aggregated in positive and negative ion modes, indicating that the detection method had good stability and repeatability, and the detection system was reliable. The blank group, model group and each treatment group could be separated, and the treatment group was between the blank group and the model group, indicating that the metabolism of POI rats tended to be normal after treatment.

[0200] The obtained metabolomics mass spectrum raw data was converted into.abf format file by Analysis Base File Converter software, and then was subjected to chromatographic peak alignment and normalization pretreatment by MS-DAIL 3.9 software and SEERF platform (https: / / slfan2013.github.io / SERRF-online / ). Then the data was introduced into SIMCA-P 14.1 software for principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA). The differential metabolites were screened according to variable importance p (VIP)>1 and P<0.05, combined with the fold change (FC) and P value of the differences between the model group and the blank group, and between the model group and the high dose group, with FC>1.2 and P<0.05 or FC<0.8333 and P<0.05 as the screening conditions, combined with human metabolome database (HMDB, https: / / hmdb.ca), and finally the potential biomarkers were confirmed. A total of 9 differential metabolites were identified in two ion modes, which were 5-methylcytosine, dibutyl phthalate, phytosphingosine, dodecyl ethanolamine, diethanolamine lauric acid, PC (16:0 / 18:1 (9Z)), LysoPC (18:2 / 0:0), LPC 16:0 and 17:0 Lyso PC.

[0201] Table 19 Endogenous differential metabolites in rat serum

[0202] 6. Clinical effect evaluation

[0203] (1) Case source

[0204] Jiangsu Provincial Hospital of Traditional Chinese Medicine

[0205] (2) Western diagnostic criteria

[0206] 1. Abdominal (gastrointestinal) surgery to remove a certain part

[0207] 2. Postoperative nausea, vomiting, unable to eat orally

[0208] 3. Stop exhaust, defecation

[0209] (3) Inclusion criteria

[0210] 1. Meet the western diagnostic criteria for intestinal paralysis after abdominal surgery for gastrointestinal cancer

[0211] 2. 18 years old ≤ age < 75 years old

[0212] 3. Sign the informed consent form and voluntarily participate in this trial

[0213] (4) Exclusion criteria

[0214] 1. History of allergic reactions to traditional Chinese medicine and major components and drug-induced liver damage

[0215] 2. Unwilling to accept traditional Chinese medicine treatment

[0216] 3. Patients with serious primary diseases of heart, liver, kidney and hematopoietic system, etc. and patients with mental illness

[0217] 4. Patients participating in other drug clinical trials

[0218] (5) Research methods

[0219] Basic treatment and other treatment orders: ERAS routine treatment + postoperative ventilation formula combined treatment

[0220] Treatment group: Postoperative ventilation formula granules, 1 bag per day, 100 mL hot water, 2 times a day, 50 mL each time

[0221] Control group: ERAS routine treatment

[0222] Course of treatment: 3 days

[0223] (6) General information

[0224] Subject name, hospital number, start date of the trial, demographic characteristics: gender, age, height, weight

[0225] Vital signs: body temperature, resting heart rate, respiration, blood pressure (systolic and diastolic) after 10 minutes of rest

[0226] (7) Observation index

[0227] Primary efficacy index: the time of first ambulation, the time of recovery of bowel sound, the time of postoperative ventilation, the time of first defecation after operation Secondary efficacy index: whether there are shortness of breath, fatigue, nausea and vomiting, dizziness and headache, diarrhea and abdominal pain, etc.

[0228] (8) Data processing

[0229] The SPSS statistical software was used for analysis, and P<0.05 was used to determine whether the difference was statistically significant.

[0230] Application Example 1

[0231] Application Example 2

[0232] Application Example 3

[0233] Application Example 4

[0234] Application Example 5

[0235] Application Example 6

[0236] Table 20 Postoperative recovery data table of patients

[0237] The postoperative ventilation time, the postoperative defecation time, and the first ambulation time of the above 6 patients were counted. After taking the postoperative ventilation recipe, the postoperative ventilation time of the above patients was shortened to 35-41 hours, the postoperative defecation time was also advanced to 3-4 days, and the patients could ambulate from the first day after operation, which indicated that the postoperative ventilation recipe could promote the postoperative intestinal function recovery of patients after abdominal surgery.

[0238] The above has been described illustratively for the present invention and its embodiments, and the description is not restrictive, so if a person skilled in the art is inspired by it, without departing from the purpose of the present invention, similar structural modes and embodiments of the technical solutions are not designed creatively, and all should belong to the protection scope of the present invention.

Claims

1. A postoperative ventilation medicinal composition, characterized in that, The Chinese medicine components are made from the following heavy number of parts: Fried Semen Raphani 15-45 parts, Fried Perilla Fruit 5-15 parts, Fried Semen Sophorae 5-15 parts, Fried Endothelium Corneum Gigeriae Galli 4.5-13.5 parts, Charred Hawthorn Fruit 5-15 parts, Tangerine Peel 3-9 parts, Herba Taraxaci 15-45 parts, and Licorice 1.5-4.5 parts.

2. The postoperative ventilation traditional Chinese medicine composition according to claim 1, characterized in that, The Chinese medicine components are made from the following heavy number of parts: Fried Semen Raphani 15-30 parts, Fried Perilla Fruit 5-10 parts, Fried Semen Sophorae 5-10 parts, Fried Endothelium Corneum Gigeriae Galli 4.5-9 parts, Charred Horthorn Fruit 5-10 parts, Tangerine Peel 3-6 parts, Herba Taraxaci 15-30 parts, and Licorice 1.5-3 parts.

3. The postoperative ventilation traditional Chinese medicine composition according to claim 2, characterized in that, The Chinese medicine components are made from the following heavy number of parts: Fried Semen Raphani 30 parts, Fried Perilla Fruit 10 parts, Fried Semen Sophorae 10 parts, Fried Endothelium Corneum Gigeriae Galli 9 parts, Charred Horthorn Fruit 10 parts, Tangerine Peel 6 parts, Herba Taraxaci 30 parts, and Licorice 3 parts.

4. The method for preparing the postoperative ventilation traditional Chinese medicine composition according to any one of claims 1-3, characterized in that, The method comprises the following steps: (1) Fried Semen Raphani, Fried Semen Sophorae, Fried Perilla Fruit, Herba Taraxaci, Fried Endothelium Corneum Gigeriae Galli, Charred Horthorn Fruit, Tangerine Peel, and Licorice are taken by weight parts, and decocted twice with water. The first time, the water is added to 12 times the volume of the decoction pieces, soaked for 35 minutes, and then heated and decocted for 40 minutes. The filtrate is collected. The second time, the water is added to 6 times the volume of the decoction pieces, heated and decocted for 30 minutes, and then filtered. The filtrate is collected. The two filtrates are combined, concentrated to 65℃ relative density 1.05-1.10 under reduced pressure, and then centrifuged by a straight tube high-speed centrifuge to obtain a centrifuged solution; (2) Take dextrin and place it in a fluidized bed. When the material temperature rises, start feeding the centrifuged solution at a speed of 80-150 r / min. Spray dry to obtain the product.

5. Use of the composition of any one of claims 1-3 in the preparation of a drug for treating postoperative ileus.

6. Use of the composition of any one of claims 1-3 in the preparation of a drug for postoperative ventilation.

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