Traditional Chinese medicine composition for treating osteomyelitis

Abstract

The invention discloses a traditional Chinese medicine composition for treating osteomyelitis. The traditional Chinese medicine composition is prepared from the following raw materials: honeysuckle stem, patrinia rugosa, raw astragalus membranaceus, spina gleditsiae, radix cyathulae, roasted rhizoma atractylodis macrocephalae, poria cocos, fructus aurantii, fructus forsythiae, angelica sinensis, ligusticum wallichii, parched white peony root, Chinese yam and coix seed. Based on pathogenesis, the treatment method is layered torsion, according to defensive qi and nutrient blood dialectical differentiation, evil is mainly dispersed in defensive qi, and fructus forsythiae is used for dispersing the evil in exterior. After losing and mistakenly treating, pathogen enters nutrient blood, collaterals are stagnated, hard lumps are formed, heat is formed by hiding in the interior, a Tongtou method is used for removing toxins, clearing heat, penetrating toxins, softening the hard lumps and dissipating binds, and spina gleditsiae, radix paeoniae alba, honeysuckle stem and patrinia rugosa are used for promoting tissue regeneration, penetrating pathogen and removing nutrient blood.

Description

Technical Field

[0001] The invention belongs to the field of traditional Chinese medicine, and particularly relates to a traditional Chinese medicine composition for treating osteomyelitis. Background Art

[0002] Osteomyelitis is an infectious disease of the bone and bone marrow that can be caused by a variety of pathogens, including aerobic or anaerobic bacteria, mycobacteria, and fungi. The disease is mostly caused by pathogens such as Staphylococcus aureus invading bone tissue through bloodstream infection, direct spread, or open fractures, triggering an inflammatory response, leading to bone destruction and sequestration. The causes of osteomyelitis are diverse, and underlying diseases such as trauma, surgery, and diabetes can increase the risk of developing the disease. Once the disease occurs, patients often experience severe pain and swelling in the affected area, accompanied by systemic symptoms such as fever. In severe cases, it can lead to limb dysfunction and even be life-threatening.

[0003] From a Western medical perspective, the pathogenesis of osteomyelitis primarily involves the invasion, colonization, and proliferation of pathogens, as well as the body's immune response. Once pathogens enter bone tissue, they rapidly multiply and release a variety of toxins and enzymes, damaging the structure and function of bone tissue. Simultaneously, the body's immune system is activated, triggering a series of inflammatory responses, such as the accumulation of white blood cells and the release of inflammatory mediators, to combat the pathogen's invasion. However, if the inflammatory response is excessive or prolonged, it can further damage bone tissue and worsen the condition.

[0004] Commonly used Western medicine treatments for osteomyelitis include antibiotics and surgical treatment. Antibiotics are the first choice for treating osteomyelitis, controlling infection by inhibiting or killing pathogens. Commonly used antibiotics include the following categories: Penicillins, such as amoxicillin, have strong bactericidal effects against Gram-positive bacteria and are suitable for treating osteomyelitis caused by bacteria such as Staphylococcus aureus. However, some patients may be allergic to them, and drug resistance may exist. Cephalosporins, such as cefazolin and cefuroxime, have broad-spectrum antimicrobial activity against a variety of Gram-positive and Gram-negative bacteria. They can be used to treat osteomyelitis caused by sensitive bacteria, but long-term use may lead to adverse reactions such as dysbiosis. Quinolones, such as levofloxacin, have good antimicrobial effects against Gram-negative and some Gram-positive bacteria and are suitable for patients who are not suitable for traditional antibiotics. However, they may cause gastrointestinal and central nervous system side effects. Glycopeptides, such as vancomycin, are primarily used to treat serious infections caused by resistant strains such as methicillin-resistant Staphylococcus aureus. However, their significant adverse reactions, such as nephrotoxicity and ototoxicity, limit their long-term use.

[0005] Surgical treatment controls the spread of infection by removing the lesion and draining pus, and is often used in conjunction with antibiotics to improve the therapeutic effect. However, surgical treatment has disadvantages such as large trauma and slow recovery, and there is still a risk of recurrence after surgery.

[0006] Compared with Western medicine, Traditional Chinese Medicine (TCM) offers unique advantages in treating osteomyelitis. Traditional Chinese Medicine (TCM) believes that osteomyelitis is often caused by insufficient vital energy and the invasion of pathogenic qi, leading to stagnation of qi and blood, and malnutrition of the bones and muscles. By strengthening vital energy, dispelling pathogenic qi, promoting blood circulation and removing stasis, and clearing away heat and detoxifying, TCM treatment can fundamentally improve the body's internal environment, enhancing immunity and disease resistance, and ultimately curing the disease. Furthermore, TCM's multi-target mechanism of action effectively combats inflammation, promotes tissue repair, and improves microcirculation, with relatively minimal adverse reactions and a high safety and tolerability profile. This provides a gentler, more durable solution for the treatment of osteomyelitis. Summary of the Invention

[0007] The purpose of the present invention is to provide a traditional Chinese medicine composition for treating osteomyelitis, in order to provide a new solution for the treatment of osteomyelitis.

[0008] In order to achieve the above object, the technical solution adopted by the present invention is: A traditional Chinese medicine composition for treating osteomyelitis is prepared from the following raw materials in parts by weight: 15-18 parts of honeysuckle vine, 15-20 parts of patrinia scabra, 20-24 parts of raw astragalus, 8-12 parts of licorice thorn, 10-15 parts of cyathula, 10-15 parts of stir-fried atractylodes, 15-18 parts of poria, 10-15 parts of fructus aurantii, 8-12 parts of forsythia, 10-15 parts of angelica, 10-15 parts of ligusticum chuanxiong, 10-15 parts of stir-fried white peony root, 8-12 parts of yam and 25-30 parts of coix seed.

[0009] In order to further realize the present invention, the Chinese medicine composition is preferably made of the following raw materials in the following weight ratios: 15 parts of honeysuckle vine, 15 parts of scutellaria baicalensis, 24 parts of raw astragalus, 10 parts of soapberry thorn, 15 parts of Sichuan cypress, 10 parts of stir-fried Atractylodes macrocephala, 15 parts of Poria, 10 parts of Citrus aurantium, 10 parts of Forsythia suspensa, 10 parts of Angelica sinensis, 10 parts of Chuanxiong, 10 parts of stir-fried white peony root, 10 parts of yam and 25 parts of coix seed.

[0010] In order to further realize the present invention, the Chinese medicine composition is preferably prepared in the form of a decoction, pills, tea, oral liquid, tablet, capsule, granule, paste or powder.

[0011] The beneficial effects of the present invention compared to the prior art are: 1. Pathogenesis Analysis of the Present Invention Traditional Chinese medicine diagnosis believes that insufficient vital energy is the pathogenesis of osteomyelitis. Osteomyelitis patients usually have deficiency of Qi and blood. Deficiency leads to Qi stagnation and blood stasis. If Qi does not flow, it is not enough to promote blood circulation, leading to stagnation and blood stasis. If the stasis lasts for a long time, the stasis will turn into heat, and then become toxic and pus. Before the formation of pus, the main symptom is the excess of toxic heat, and after the formation of pus, the main symptom is deficiency of Qi, blood and vital energy.

[0012] Clinical manifestations of chronic osteomyelitis due to Qi and blood deficiency include: pale complexion, fatigue, spontaneous sweating, decreased appetite, palpitations, shortness of breath, clear, thin sinus discharge, floating grayish-white granulation tissue, poor healing, a pale red tongue with a thin white coating, and a thready or weak pulse. This syndrome is more common in the late stages, with X-rays often revealing dead bone. Prolonged sinus healing leads to progressive depletion of Qi and blood, resulting in a weak body unable to expel toxins.

[0013] The Suwen (Suwen) Treatise on Acupuncture states: "When righteous qi exists within, evil cannot invade." Whether the human body is susceptible to evil qi and becomes ill in daily life, as well as the speed of disease progression, manifestations, and outcomes, all depend, to a certain extent, on the sufficiency of righteous qi. In our long-term treatment of osteomyelitis, we have found that the spleen and stomach are the foundation of acquired constitution. A healthy spleen and stomach balances qi and blood, preventing internal stagnation and external invasion by the six evil pathogens. Where evil pathogens gather, qi must be deficient. This fundamentally clarifies the internal factors that cause and develop disease and is of great significance for understanding the pathogenesis of sepsis. For example, elderly patients with multiple underlying conditions (such as hypertension, diabetes, and hyperlipidemia) and chronic illness weaken their bodies, leading to a state of righteous qi deficiency and imbalance between yin and yang. External pathogens can easily exploit this vulnerability and, without internal righteous qi to resist, wreak havoc. In the human body, qi functions externally to strengthen the exterior and resist evil pathogens, while qi functions internally to maintain the internal organs and promote qi and blood circulation. Severe underlying diseases, trauma, major surgery, etc. can all cause massive bleeding or concurrent infection, leading to dysfunction of the body's internal organs and abnormal distribution of qi, blood, and body fluids. These factors form the pathological basis of insufficient vital energy in chronic osteomyelitis and are also an important pathogenesis of osteomyelitis.

[0014] 2. Analysis of the prescription of the present invention Based on the pathogenesis, treatment should be layered and reversed. Based on the differentiation of Wei Qi, Ying Qi, and Blood, if the pathogen is in the Wei Qi, the primary approach is to dissipate and disperse it. Forsythia suspensa can dissipate the pathogen on the surface. If the pathogen enters the Ying and Blood, causing stasis in the collaterals, forming a hard mass, which then accumulates and transforms into heat. Treatment should be based on the method of dispersing and purging to expel the poison, clearing away heat and toxins, softening and dispersing the mass. For this, use Sophora flavescens, White Peony Root, Honeysuckle Vine, and Patrinia scabra to dissipate the pathogen inside and promote tissue regeneration, allowing the pathogen to exit the Ying and Blood.

[0015] The present invention discloses the monarch medicine of the present invention comprises raw astragalus and angelica. Raw astragalus replenishes qi and strengthens the exterior, drives out toxins, promotes diuresis and reduces swelling, is an important medicine for replenishing qi, is used for exterior deficiency and instability caused by qi deficiency, and has a leading therapeutic effect on the body weakness of osteomyelitis. Astragalus is a representative of qi-invigorating medicine, has the effects of replenishing qi and strengthening the exterior, driving out toxins and promoting tissue regeneration, and promoting diuresis and reducing swelling. For the treatment of osteomyelitis, which requires strengthening the body's resistance and promoting wound healing, raw astragalus is very suitable as the monarch medicine, has the effect of strengthening the body's resistance, helps to prevent postoperative infection, and improves the ability to resist pathogens; angelica replenishes blood, promotes blood circulation and removes blood stasis, and is combined with raw astragalus to enhance the effect of nourishing blood and promoting blood circulation, and has an auxiliary therapeutic effect on local pain and swelling caused by blood stasis. Angelica replenishes blood, promotes blood circulation and removes blood stasis, and assists raw astragalus in strengthening the effects of nourishing blood and promoting blood circulation and removing blood stasis, which is very important for osteomyelitis, which requires improving blood circulation and promoting wound healing. It complements the qi-invigorating and blood-nourishing effects of raw astragalus, helps to improve the body's repair ability, and promotes the healing of postoperative wounds.

[0016] The ministerial drugs of the present invention are honeysuckle vine, soapberry thorn and Sichuan cypress. Honeysuckle vine clears away heat and detoxifies, dispels wind and heat, and helps the monarch drug to strengthen the heat-clearing and detoxifying effect. For the heat and toxicity accumulation in osteomyelitis, the heat-clearing and detoxifying effects of honeysuckle vine and Patrinia corylifolia synergistically help to alleviate postoperative inflammatory response and relieve local redness, swelling, heat and pain; soapberry thorn reduces swelling and disperses stagnation, helps the monarch drug to detoxify and regenerate tissue, and is used to treat the symptom that local blood stasis and swelling are difficult to dissipate after surgery; Sichuan cypress promotes blood circulation and removes blood stasis, guides the drug downward, helps the monarch drug to make the medicinal effect directly reach the diseased area, and enhances the local therapeutic effect.

[0017] The adjuvants of the present invention are Patrinia corylifolia, stir-fried Atractylodes macrocephala, Poria cocos, Chuanxiong rhizome, stir-fried white peony root, Chinese yam and coix seed. Patrinia corylifolia has the effect of clearing away heat and detoxifying, helping the main drug to expel poison and promote tissue regeneration, and also helps to reduce postoperative inflammatory reaction and relieve local redness, swelling, heat and pain; stir-fried Atractylodes macrocephala strengthens the spleen and eliminates dampness, helping the main drug to strengthen qi and blood nourishment, targeting at the bone carbuncle caused by spleen deficiency and dampness, blood deficiency and poor nourishment, slow wound healing, regulating the acquired foundation, and promoting postoperative recovery of patients; Poria cocos promotes water and dampness, strengthens the spleen and calms the mind, helps stir-fried Atractylodes macrocephala to eliminate dampness, targeting at the poor wound healing caused by internal dampness, unblocks the water channel, and promotes postoperative recovery of spleen and stomach function of patients; Chuanxiong promotes blood circulation and removes blood stasis , promotes qi and relieves pain, assists Sichuan Achyranthes bidentata in promoting blood circulation and removing blood stasis, and together with white peony root, assists the monarch drug Angelica sinensis to enhance the effect of promoting blood circulation and removing blood stasis, helps to improve local blood circulation, and reduces the formation of hematoma and blood stasis after surgery; fried white peony root nourishes blood and softens the liver, assists the monarch drug Angelica sinensis in nourishing blood, relieves acute pain, and improves the pain and discomfort symptoms of postoperative patients; yam nourishes the spleen and kidneys, and targets the postoperative physical weakness of osteomyelitis caused by deficiency of both spleen and kidney, and helps the monarch drug to replenish qi and blood; coix seed promotes water and dampness, strengthens the spleen and stops diarrhea, and together with Poria cocos and Atractylodes macrocephala, it strengthens the spleen and removes dampness, replenishes qi and replenishes deficiency, and targets the symptoms of excessive exudation and slow wound healing after surgery caused by dampness.

[0018] The guiding drugs of the present invention are Forsythia suspensa and Citrus aurantium. Forsythia suspensa, as a guiding drug, guides other drugs to directly reach the diseased area, thereby enhancing the overall effect of the prescription. At the same time, it itself has the effects of clearing away heat and detoxifying, dispersing nodules and reducing swelling, and is known as the "holy medicine for sores". It helps the main drugs, together with Honeysuckle, Patrinia scabra, and Gleditsia sinensis, to expel toxins and promote tissue regeneration, and is clinically used to control infection after surgery; Citrus aurantium regulates qi and relieves fullness, and is used to treat symptoms such as chest tightness caused by qi stagnation. As a guiding drug, it can regulate qi and relieve fullness, coordinate the effects of other drugs in the prescription, make the efficacy more coordinated, reduce adverse reactions, regulate qi, prevent Forsythia suspensa, Patrinia scabra, Gleditsia sinensis, Honeysuckle and other drugs from causing cold stagnation and qi stagnation, and help Atractylodes macrocephala, Poria cocos, Chinese yam, and Coix seed to strengthen the spleen, promote qi and eliminate dampness. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 This is a picture showing the state changes at each stage during the treatment of typical chronic osteomyelitis using the Chinese medicine composition of the present invention in Experimental Example 1 of the present invention; Figure 2 HE staining of the toxicity results of Experimental Example 2 of the present invention (the scale bar in the figure is 50 μm); Figure 3 The results of routine blood test analysis of the toxic effect results of Experimental Example 2 of the present invention (* indicates the comparison with the Control group, p < 0.05); Figure 4 The biochemical analysis results of the toxic effect results of Experimental Example 2 of the present invention (* indicates a comparison with the Control group, p < 0.05); Figure 5 The test results of Micro-CT in Experimental Example 2 of the present invention are shown in the figure (the scale is 2 mm); Figure 6 The results of ELISA analysis in Experimental Example 2 of the present invention are shown in Table 2 (* indicates significant difference, *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001); Figure 7 This is the HE staining image of the tibial tissue in Experimental Example 2 of the present invention; Figure 8 This is a diagram showing the WB test results of tibial tissue in Experimental Example 2 of the present invention; Figure 9 These are the WB analysis results of tibial tissue in Experimental Example 2 of the present invention (* indicates significant differences, *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001). DETAILED DESCRIPTION

[0020] The present invention will be further described below with reference to specific implementation methods, clinical trials and animal experiments.

[0021] Example 1: Honeysuckle vine 15g, Patrinia corylifolia 15g, raw Astragalus 20g, Sophora flavescens 8g, Cyathula 10g, stir-fried Atractylodes 10g, Poria 15g, Citrus aurantium 10g, Forsythia suspensa 8g, Angelica 10g, Ligusticum chuanxiong 10g, stir-fried White Peony 10g, Chinese Yam 8g and Coix seed 25g.

[0022] Example 2: Honeysuckle vine 18g, Patrinia corylifolia 20g, raw Astragalus 24g, Sophora flavescens 12g, Cyathula 15g, stir-fried Atractylodes 15g, Poria 18g, Citrus aurantium 15g, Forsythia suspensa 12g, Angelica 15g, Chuanxiong 10-15g, stir-fried White Peony 15g, Chinese Yam 12g and Coix seed 30g.

[0023] Example 3: Honeysuckle vine 15g, Patrinia corylifolia 15g, raw Astragalus 24g, Sophora flavescens 10g, Cyathula 15g, stir-fried Atractylodes 10g, Poria 15g, Citrus aurantium 10g, Forsythia suspensa 10g, Chinese angelica 10g, Chuanxiong 10g, stir-fried White Peony 10g, Chinese yam 10g and Coix seed 25g.

[0024] The Chinese medicine composition is preferably prepared in the form of a decoction, pill, tea, oral liquid, tablet, capsule, granule, paste or powder.

[0025] Experimental Example 1: Clinical Trial 1 Materials and Methods 1.1 Research subjects and groups The clinical study included patients with post-traumatic tibial osteomyelitis due to Qi and blood deficiency who were treated at Gansu Provincial Hospital of Traditional Chinese Medicine from September 2017 to December 2020. All patients were randomly divided into an experimental group and a control group. The control group received conventional bone transport, while the experimental group received perioperative oral administration of Shengji Bugu Gao combined with bone transport. The inventors' team explained the study content and procedures in detail to all patients and their families. All patients voluntarily participated in the study, and the inventors' team, patients, and their families signed informed consent forms.

[0026] 1.2 Study inclusion and exclusion criteria Inclusion criteria: (1) age 18 years or older; (2) unilateral involvement, secondary osteomyelitis due to infection after traumatic tibial fracture, meeting the diagnostic criteria of Cierny-Mader type IV chronic osteomyelitis; (3) mild to moderate anemia after bone transfer surgery, with hemoglobin level between 75-110 g / L; (4) syndrome of qi and blood deficiency according to traditional Chinese medicine; 5) good patient compliance, completion of clinical follow-up, and complete data.

[0027] Exclusion criteria: (1) under 18 years old; (2) combined with fractures or organ injuries in other parts of the body; (3) combined with serious underlying medical diseases, such as severe liver and kidney dysfunction, cardiovascular disease, severe anemia, uncontrolled diabetes and other endocrine diseases; (4) abnormal coagulation function or chronic wasting disease; (5) malignant tumors and advanced tumors; (6) severe mental or psychological diseases and unwilling to cooperate with examinations and follow-up.

[0028] 1.3 Intervention plan 1.3.1 Control group The control group received traditional bone transport. The main steps were: (1) Preoperative preparation: assess local tissue necrosis, conduct imaging examinations, and select sensitive antibiotics. (2) Thorough debridement: use VSD negative pressure drainage or flap surgery to cover if necessary. (3) Installation of Ilizarov ring external fixator: two rings at the proximal and distal ends, two 2mm Kirschner wires are inserted into the upper and lower tibia, and cross-fixed on the steel rings. The middle transport ring is fixed with a half-needle and a 2.5mm Kirschner wire. (4) Osteotomy: For patients with mild infection, osteotomy can be performed in one stage; for patients with severe infection and poor general condition, or bone defects that need to be transported from distal to proximal, osteotomy can be performed in two stages 4-6 weeks after the first surgery. (5) Wound repair: Most patients can undergo direct primary suture, and some patients need to combine flap repair with wound repair. (6) Postoperative treatment: use sensitive antibiotics and elevate the affected limb. (7) Bone transport: bone transport begins one week after osteotomy. The adjustment speed is 1mm / d, completed in 4 times, each time extending 0.25mm, i.e. 1 / 4 circle. (8) Functional exercise: On the second day after surgery, quadriceps isometric contraction is started, and ankle plantar flexion and dorsiflexion exercises are guided.

[0029] 1.3.2 Experimental Group In addition to receiving the same bone transfer treatment as the control group, the patients in the experimental group took the decoction prepared in Example 3 of the Chinese medicine composition of the present invention (prepared uniformly by the preparation room of Gansu Provincial Hospital of Traditional Chinese Medicine, including raw astragalus, honeysuckle, platycodon, licorice, chuanxiong, jujube, stir-fried atractylodes, poria, fructus aurantii, licorice, forsythia, angelica, chuanxiong, stir-fried white peony root, yam, coix seed, etc.) during the perioperative period. The dosage was 200 ml / bag. The oral administration frequency during the perioperative period was 200 ml / time, twice a day. The oral administration time points were: (1) oral administration one day before surgery; (2) oral administration starting from the fourth day after surgery for three consecutive weeks; (3) oral administration starting from the third month after surgery for three consecutive weeks, once every two months, for a total of three times.

[0030] 1.4 Main outcome measures Infection indicators, including white blood cell count, erythrocyte sedimentation rate, and C-reactive protein (CRP) levels, were detected in all patients before surgery and on days 2, 7, 14, and 25 after surgery.

[0031] 1.5 Secondary Observation Indicators 1.5.1 Short-term indicators Blood loss indicators, including red blood cell count and hemoglobin, were measured in all patients before surgery and on days 2, 7, 14, and 25 after surgery.

[0032] 1.5.2 Long-term indicators Follow-up visits were conducted every two months after surgery to observe the following clinical parameters: (1) Local wound healing and signs of reinfection. (2) X-rays were taken to assess bone healing, record bone healing time, and assess whether poor healing or delayed healing occurred. (3) External fixation time was recorded, and the external fixation index was calculated, defined as the external fixation time per unit bone extension distance. Bone transport rate was calculated, defined as the time required for unit bone extension distance. (4) Visual Analogue Score (VAS) was used to evaluate pain on the 25th day after surgery (i.e., 3 weeks after medication).

[0033] The ankle and knee function of all patients were evaluated before and after surgery. The evaluation was as follows: (1) Ankle function was assessed using the AOFAS Ankle Hindfoot Scale. This scale evaluates pain, function and voluntary movement, walking, foot stability, and alignment, with a maximum score of 100. (2) Knee function was assessed using the Hospital for Special Surgery (HSS) score. This scale evaluates pain, function, range of motion, muscle strength, knee flexion deformity, and knee instability, with a maximum score of 100.

[0034] The clinical efficacy of the two groups was evaluated and compared. The criteria were as follows: (1) 25 days after surgery, the syndrome differentiation criteria and symptom and sign grading quantitative criteria for Qi and blood deficiency syndrome were formulated in accordance with the "Guiding Principles for Clinical Research of New Chinese Medicine Drugs", and the TCM symptom score was obtained, with a full score of 20 points. (2) After postoperative bone healing, the patients were divided into four grades: excellent, good, acceptable, and poor according to the Johner-Wruhs tibial shaft efficacy evaluation criteria. The excellent and good rate was calculated as (excellent + good) / (excellent + good + acceptable + poor) × 100%.

[0035] The complications that occurred in the two groups of patients during the follow-up period were observed, including wound infection, nail tract infection, poor healing of the butt joint, recurrence of osteomyelitis, foot drop and ankle stiffness.

[0036] 1.6 Statistical analysis The inventors' team used SPSS version 21.0 statistical software for data analysis. Enumeration data were expressed as number of cases (n) and percentage (%), and measurement data were expressed as mean ± standard deviation (x ± s). Enumeration data were compared using the chi-square test or Fisher's exact test. Measurement data that followed a normal distribution were tested using the t-test, while those that did not follow a normal distribution were tested using the signed-rank test. An α value of 0.05 was used as the test level, and a P value of < 0.05 was considered statistically significant.

[0037] 2. Results 2.1 General Information The clinical study initially enrolled 86 eligible patients with Qi and blood deficiency-related osteomyelitis. Four patients were lost to follow-up, and two withdrew from the study, ultimately enrolling a total of 80 patients. The experimental group included 23 males and 17 females with a mean age of 42.6 ± 10.3 years (range, 20-65 years). The control group included 24 males and 16 females with a mean age of 42.1 ± 11.2 years (range, 19-62 years). The primary injury types in the experimental group included 31 open injuries and 9 closed injuries. The initial causes of injury were traffic accidents (16), falls (10), heavy object injuries (9), and other causes (5). The control group included 32 open injuries and 8 closed injuries. The initial causes were traffic accidents (14), falls (11), heavy object injuries (11), and other causes (4). The mean disease duration in the experimental group was 14.6 ± 5.2 months (range, 5–28 months). Twenty-four patients underwent multiple debridement and flap procedures, 30 patients developed skin and soft tissue defects ranging in size from 2 cm × 3 cm to 10 cm × 17 cm, and 22 patients developed sinus sequestrum with purulent drainage. The mean disease duration in the control group was 13.9 ± 5.1 months (range, 6–29 months). Twenty-two patients underwent multiple debridement and flap procedures, 29 patients developed skin and soft tissue defects ranging in size from 3 cm × 3 cm to 12 cm × 18 cm, and 20 patients developed sinus sequestrum with purulent drainage. The mean bone defect length after debridement and osteotomy in the experimental and control groups was 7.3 ± 2.6 cm and 7.1 ± 2.4 cm, respectively. The surgical bleeding volume of the experimental group and the control group were (147.3±32.1) ml and (135.6±29.5) ml, respectively. The intraoperative blood transfusion volume was (80.4±23.6) ml and (78.2±19.5) ml, respectively. There was no significant difference in the baseline data between the two groups ( P >0.05), see Table 1 for details.

[0038]

[0039] 2.2 Short-term indicators There were no significant differences in preoperative red blood cell count, hemoglobin, white blood cell count, CRP, and erythrocyte sedimentation rate between the two groups (P>0.05). Twenty-five days after surgery, the red blood cell count (P=0.027) and hemoglobin level (P=0.015) in the experimental group were significantly higher than those in the control group, while the white blood cell count (P<0.001), CRP (P=0.008), and erythrocyte sedimentation rate (P=0.019) were significantly lower than those in the control group. Comparison of laboratory parameters between the two groups is shown in Table 2.

[0040]

[0041] 2.3 Long-term indicators 2.3.1 Clinical indicators Both groups were followed up for an average of 17.1±4.2 months and 18.2±4.3 months, respectively. The skin healed well in both groups, and functional exercises were performed postoperatively. Primary union of the fracture ends and the extension zone was achieved in 37 patients (92.5%) in the experimental group and 31 patients (77.5%) in the control group. The remaining patients experienced nonunion or poor osteogenesis, which healed after secondary bone grafting. The mean bone healing time (256.7±59.1 vs 328.5±74.6 d, P<0.001) and external fixation index (34.1±7.2 vs 40.3±8.6 d / cm, P=0.013) in the experimental group were significantly shorter than those in the control group. The bone turnover rate was significantly higher in the experimental group (11.4±3.5 vs 8.2±3.1 d / cm, P<0.001), and the postoperative VAS score was significantly lower than that in the control group (3.1±1.2 vs 5.9±2.5, P=0.011). The comparison of clinical indicators between the two groups is shown in Table 3.

[0042]

[0043] 2.3.2 Joint function indicators There was no significant difference in the ankle AOFAS score and knee HSS score between the two groups before surgery (P>0.05). The joint function was significantly improved after treatment, and the AOFAS score (91.3±10.6 vs 83.5±9.4, P<0.001) and HSS score (87.4±5.9 vs 77.5±6.1, P<0.001) of the experimental group were significantly higher than those of the control group. The specific results are shown in Table 4.

[0044]

[0045] 2.3.3 Clinical efficacy 25 days after surgery, the average TCM symptom score of the experimental group was significantly higher than that of the control group (16.3±4.7 vs 10.5±3.9, P<0.001). The results of the Johner-Wruhs tibial shaft efficacy evaluation of the two groups of patients are shown in Table 5. The experimental group included 24 excellent cases, 14 good cases, and 2 acceptable cases, with an overall excellent and good rate of 95.0%; the control group included 16 excellent cases, 12 good cases, 9 acceptable cases, and 3 poor cases, with an overall excellent and good rate of 70.0%. Typical case pictures are shown in Figure 1 , Figure 1 It includes stages such as preoperative preparation, surgical operation, postoperative treatment and follow-up, reflecting the overall treatment process and the changes in the patient's status at each stage.

[0046]

[0047] Figure 1 A 46-year-old female patient developed post-traumatic osteomyelitis of the tibia one year after internal fixation for a tibial fracture. Preoperative anteroposterior (a) and lateral (b) radiographs show a localized periosteal reaction; a preoperative gross radiograph (c) demonstrates localized sinus formation with purulent drainage; intraoperative photographs (d, e) show removal of the original internal fixation device, debridement and resection of the large dead bone in the lesion, and tibial metaphyseal osteotomy. Postoperative gross radiograph (f) shows the installation of an Ilizarov circular external fixator and negative pressure suction of the VSD wound surface; 1-week postoperative anteroposterior (g) and lateral (h) X-ray films show that the external fixator was stably and firmly installed before bone transport; 9-month postoperative anteroposterior (i) and lateral (j) X-ray films show good ossification in the bone extension area; 30-month postoperative anteroposterior (k) and lateral (l) X-ray films show healing of the bone ends; 30-month postoperative gross radiographs of the affected limb in anteroposterior (m), lateral (n) and flexion positions (o) show complete healing of the local wound surface and good joint function.

[0048] 2.3.4 Complications During the follow-up period, the experimental group experienced one case of wound infection, three cases of nail tract infection, and one case of poor buttock healing. The control group experienced two cases of wound infection, five cases of nail tract infection, two cases of poor buttock healing, one case of foot drop, one case of ankle stiffness, and two cases of recurrent osteomyelitis. The complication rate in the experimental group was significantly lower than that in the control group (12.5% ​​vs 32.5%, P < 0.001).

[0049] Experimental Example 2: Animal Testing 1 Experimental Materials and Methods 1.1 Animal husbandry and handling Male and female Sprague-Dawley rats (8 weeks old, Beijing Huafukang Biotechnology Co., Ltd. / Beijing Biotechnology Co., Ltd., License Nos. SCXK (Beijing) 2024-0003 / SCXK (Beijing) 2024-0001) were housed at 20-26°C and 40%-70% humidity. After one week of acclimatization, the rats were divided into groups of six: 1) a blank group, 2) a low-dose group (18.27 g / kg) of the Chinese herbal composition of the present invention (TCM), 3) a medium-dose group (21.294 g / kg), and 4) a high-dose group (24.759 g / kg). The rats were gavaged with different doses of TCM once daily for 15 consecutive days. During the dosing period, the animals were weighed daily. After dosing, the rats were weighed and sacrificed, and their major organs (heart, liver, spleen, lungs, and kidneys) were removed for examination. Then, the groups were divided into 6 groups, (1) sham operation group, (2) osteomyelitis model group, (3) model + high-dose Chinese medicine group, (4) model + high-dose Chinese medicine + lesion removal group, (5) model + positive drug group and (6) model + high-dose Chinese medicine + positive drug group. The osteomyelitis model was constructed: after the animals were anesthetized, the limbs were fixed and the surgical operation was performed. A 1 cm long skin incision was made along the inner side of the anterior tibial crest below the knee joint of the right hind limb of the rat. The inner side of the tibialis anterior muscle was bluntly separated to reach the periosteum, and the tibial crest on the anterior and lateral side of the upper end of the tibia was exposed. A 1 mm diameter needle was used to drill a bone hole. The bone hole was flushed with saline. An L-shaped steel nail soaked with Staphylococcus aureus was placed into the tibial medullary cavity. After repeated flushing with saline, the wound was sutured layer by layer. After surgery, the rats were kept in a single cage. Four weeks after modeling, one rat in the model group underwent X-ray to confirm the success of the model. One rat in each of the sham operation group and the model group underwent HE staining to verify the model. After model establishment, drug administration began in each group. The high-dose Chinese medicine group (administered daily by gavage for four weeks) and the positive drug group (levofloxacin, 40 mg / kg, administered every two days for four weeks) received their corresponding drugs by gavage. The lesion debridement group underwent lesion debridement one week after model establishment: the contaminated Kirschner wire was removed, and the surrounding lesion was gently scraped. After scraping, the lesion was rinsed with saline. The Kirschner wire was replaced after visual inspection to confirm the absence of necrotic muscle and bone tissue. The animals were then sacrificed to collect blood samples and bone tissue for downstream testing.

[0050] 1.2 HE staining Tissue sections were baked, dewaxed, and hydrated, then stained with hematoxylin (ZLI-9610, Zhongshan Jinqiao) for 3-5 min. The sections were rinsed with running water, differentiated with 1% hydrochloric acid alcohol, bluing solution, and stained with eosin (G1100, Solarbio) for 3-5 min. The sections were then dehydrated, mounted, and observed under a microscope (BX43, Olympus).

[0051] 1.3 ELISA and biochemical testing ELISA kits were used to detect the changes in the levels of TNF-α (MM-0180R1, Jiangsu Enzyme Immunity) and IL-6 (MM-0190R1, Jiangsu Enzyme Immunity), or biochemical detection kits were used to detect the levels of aspartate aminotransferase (AST) (701138, Shandong Boco Biotechnology), alanine aminotransferase (ALT) (70111, Shandong Boco Biotechnology), albumin (ALB) (70113, Shandong Boco Biotechnology), total protein (TP) (70112, Shandong Boco Biotechnology) and AST. The samples were processed and tested according to the instructions to detect changes in the levels of serum creatinine (CREA) (70127, Shandong Boko Biological), urea (UREA) (70124, Shandong Boko Biological), uric acid (UA) (70125, Shandong Boko Biological), and urine microalbumin (MALB) (70128, Shandong Boko Biological).

[0052] 1.4 Immunohistochemistry Rat tibia tissue was decalcified and sectioned. After baking, dewaxing, and hydration, antigen retrieval was performed using citrate buffer. After blocking with 5% BSA, sections were incubated with ICAM1 (Affinity, AF6088, 1 / 100), CTLA4 (ABclonal, A2063, 1 / 100), TIMP1 (Affinity, AF7007, 1 / 200), and XCL1 (Immunoway, YT5307, 1 / 200) overnight at 4°C. The sections were then incubated with horseradish peroxidase-conjugated goat anti-rabbit secondary antibodies (ZB-2301, Zhongshan Jinqiao, 1 / 100), developed with DAB (CW0125M, CWBIO), and counterstained with hematoxylin for 3 minutes. After dehydration and clearing, the sections were mounted and observed under a microscope (BX43, Olympus).

[0053] 1.5 Western blotting Rat tibia tissue was collected and lysed with RIPA buffer (C1053, Beijing Pulilai Gene Technology Co., Ltd.) using a tissue grinder. Total protein was extracted and centrifuged at 12,000 rpm in a high-speed centrifuge (5424R, Eppendorf) for 10 minutes at 4°C. The supernatant was collected and quantified using a BCA protein quantification kit (E-BC-K318-M, Elabscience). After denaturation, the protein sample was subjected to sodium dodecyl sulfate gel electrophoresis (SDS-PAGE) for 1.5 hours, followed by transfer to the membrane at a constant current of 300 mA for 1 hour. A PVDF membrane (IPVH00010, Millipore) was blocked with skim milk powder and incubated with the primary antibody overnight at 4°C. The next day, the membrane was incubated with the secondary antibody for 2 hours at room temperature. The membrane was then soaked with ultrasensitive luminescent solution (RJ239676, Thermo Fisher Scientific) and visualized using an ultra-high-sensitivity chemiluminescence imaging system (Tanon-5200, Shanghai Tianneng Technology Co., Ltd.). The antibodies used and the corresponding dilution factors are shown in Table 6:

[0054] 1.6 Statistical analysis Graphpad Prism 9.0 software was used for graphics and statistical analysis. All experiments were repeated three times, and quantitative results are expressed as mean ± standard deviation (X ± S). Quantitative values ​​between groups were compared using one-way analysis of variance, and pairwise comparisons were performed using the Tukey method. P < 0.05 was considered significant.

[0055] 2. Results 2.1 Toxic effects of the Chinese medicine composition of the present invention Rats were gavaged with different doses of the herbal medicine (low dose 18.27 g / kg, medium dose 21.294 g / kg, and high dose 24.759 g / kg) once daily for 15 consecutive days. Following administration, major organs (heart, liver, spleen, lungs, and kidneys) were harvested for hematology (HE) staining, whole blood was collected for routine blood tests, and serum was collected for biochemical analysis (AST and ALT, creatinine, and urea nitrogen). Figure 2-4 The results of the toxic effects of the Chinese medicine composition of the present invention on rats were obtained by HE staining to observe the changes in the tissue structure of the heart, liver, spleen, lung, kidney and other major organs of rats in different dose groups. It can be seen that the high-dose group had no obvious damage to the animal body. The key indicators such as white blood cell count, neutrophil percentage, lymphocyte percentage, monocyte count, red blood cell count and hemoglobin were detected. The results showed that the platelet count in the low-dose gavage group showed a significant downward trend. The liver function indicators ALT, AST, creatinine, urea nitrogen, etc. were detected. It was found that the AST in the low-dose gavage group showed a significant downward trend, and there were no significant differences in other indicators between the groups.

[0056] like Figure 2As shown, under an optical microscope, the control group showed slight cardiac structural changes, with more regular myocardial arrangement, distinct myocardial texture, and minor hemorrhages in the interstitial space (green arrows). The low-dose group showed slight cardiac structural changes, with more regular myocardial arrangement, distinct myocardial texture, and minor hemorrhages in the interstitial space (green arrows). The medium-dose group showed slight cardiac structural changes, with more regular myocardial arrangement, distinct myocardial texture, and minor hemorrhages in the interstitial space (green arrows). The high-dose group showed enlarged interstitial spaces between myocardial fibers, with minor hemorrhages in the interstitial spaces (green arrows). The control group showed normal liver tissue structure, with regular hepatocytes and no obvious pathological changes. The low-dose group showed normal liver tissue structure, with regular hepatocytes and no obvious pathological changes. The medium-dose group showed normal liver tissue structure, with minor hemorrhages in the interstitial space (green arrows). The high-dose group showed disorganized hepatocytes and increased hemorrhages in the interstitial space (green arrows). The control group showed normal spleen structure, with clear demarcation between the red and white pulp, and no pathological changes. The low-dose group showed normal spleen structure, with clear demarcation between the red and white pulp, and no pathological changes. The spleen in the medium-dose group showed normal structure, with a clear demarcation between the red and white pulps, and no pathological changes were observed. The red and white pulp structures of the spleen in the high-dose group were disorganized, with unclear demarcation. The lung tissue structure in the control group was relatively normal, with intact alveoli, mild alveolar wall thickening in a few cases (yellow arrows), and slightly dilated cysts. No other significant pathological changes were observed. The lung tissue structure in the low-dose group was disorganized, with thickened alveolar walls (yellow arrows) and dilated cysts. The lung tissue structure in the medium-dose group was disorganized, with thickened alveolar walls (yellow arrows) and dilated cysts. The lung tissue structure in the high-dose group was disorganized, with thickened alveolar walls, dilated cysts, and inflammatory cell infiltration in the tubular lumen (black arrows). The kidney tissue structure in the control group was intact and normal, with regular glomeruli, tightly packed tubules, and no abnormalities in the renal interstitium. The kidney tissue structure in the low-dose group was intact and normal, with regular glomeruli, tightly packed tubules, and no abnormalities in the renal interstitium. In the medium-dose group, the renal tissue structure was intact, with normal morphology, regular glomeruli, slightly dilated renal tubules, and a small amount of vascular congestion in the renal interstitium (green arrows). In the high-dose group, the renal tissue structure was intact, with normal morphology, regular glomeruli, but with increased tubular dilation and increased renal interstitial hemorrhage (green arrows).

[0057] like Figure 3 As shown in the data, there was no significant difference in the key indicators of blood routine, including white blood cell count (WBC), neutrophil percentage (Neu), lymphocyte percentage (Lym), monocyte count (Mon), red blood cell count (RBC) and hemoglobin (HGB) between the control group and the low, medium and high dose groups of oral gavage drugs (P>0.05). Only the platelet count (PLT) showed a significant downward trend in the low dose oral gavage group compared with the control group.

[0058] like Figure 4As shown, there was no significant difference in liver function indicators ALT, CREA, and UREA between the control group and the low-, medium-, and high-dose gavage drug groups (P>0.05). Only AST showed a significant downward trend in the low-dose gavage group compared with the control group.

[0059] From the above results, it can be concluded that a high dose of the Chinese medicine composition of the present invention does not cause significant damage to the animal body, and this dose was selected for subsequent experiments.

[0060] 2.2 Micro-CT results Osteomyelitis models were constructed and divided into the following groups: 1) sham operation group, 2) osteomyelitis model group, 3) model + high-dose Chinese medicine group, 4) model + high-dose Chinese medicine + lesion clearance group, 5) model + positive drug group, 6) model + high-dose Chinese medicine + positive drug group. After the intervention, one tibia was taken from each group for ex vivo Micro-CT. The results are shown in the figure. Figure 5 As shown in Table 7, Figure 5 The data show the bone density, trabecular number, bone volume fraction and trabecular separation of different groups. The inflammatory wounds in the model group were more severe, and the Chinese medicine group showed certain healing compared with the model group. The wound surface of the Chinese medicine + lesion removal group was smaller than that of the Chinese medicine group alone. The bone healing of the positive drug group was also better. The healing of the wound surface of the Chinese medicine + positive drug group was not ideal.

[0061]

[0062] 2.3 ELISA test ELISA was used to detect the levels of inflammatory factors TNF-α and IL-6 in the serum of rats in each group. Figure 6 As shown, compared with the control group, the levels of inflammatory factors TNF-α and IL-6 were significantly upregulated in the model group. After oral administration of the Chinese medicinal composition of the present invention, the levels of TNF-α and IL-6 were significantly reduced. Furthermore, after administration of a positive drug, the levels of TNF-α and IL-6 were significantly reduced. Furthermore, after treatment with the Chinese medicinal composition of the present invention combined with a positive drug, the levels of TNF-α and IL-6 were significantly reduced. However, after treatment with the Chinese medicinal composition of the present invention combined with lesion removal surgery, the levels of TNF-α and IL-6 were reduced, but without statistical significance (P>0.05). These results indicate that the Chinese medicinal composition of the present invention can alleviate the symptoms of osteomyelitis by reducing the inflammatory response.

[0063] 2.4 HE staining like Figure 7As shown, microscopic observations of the tibial tissue structure in the sham group showed normal tibial tissue structure, clear trabecular structure, and no obvious pathological inflammatory changes. In the model group, the tibial cartilage surface was irregular, with incomplete tide lines and severe damage (black arrows). Chondrocytes were morphologically abnormal, atrophic, and even necrotic, with a significantly reduced number (red arrows), indicating severe pathological damage. In the model + high-dose Chinese medicine group, the tibial cartilage surface was irregular, with mild matrix degeneration (black arrows). Chondrocytes were unevenly distributed within the matrix, with varying morphology and size (red arrows). In the model + high-dose Chinese medicine + focus debridement group, the tibial cartilage surface was relatively intact, with a loose matrix (black arrows) and varying chondrocyte morphology and size (red arrows). In the model + positive drug group, the tibial cartilage surface was relatively intact, with a loose matrix (black arrows) and varying chondrocyte morphology and size (red arrows). In the model + high-dose Chinese medicine + positive drug group, the tibial cartilage surface was relatively intact, with a loose matrix (black arrow), uneven distribution of chondrocytes within the matrix, and an increase in localized cell numbers and varying cell sizes (red arrow). In summary, compared with the model group, drug treatment showed improvement in lesions, demonstrating a therapeutic effect.

[0064] 2.5 Western blotting The protein expression levels of TGF-β1, Smad2 / 3, BMP2, p-Smad2 / 3, VEGF, and MMP-9 in the tibia tissue of rats in each group were detected by WB. Figure 8-9 As shown in the results, compared with the control group, the expression of TGF-β1, Smad2 / 3, p-Smad2 / 3, BMP2, and MMP-9 in the model group was significantly decreased, while VEGFA decreased in the model group but without statistical significance (P>0.05); compared with the model group, the expression of TGF-β1, Smad2 / 3, BMMP2, MMP-9, and VEGFA in the model + high-dose Chinese medicine group was significantly increased, while p-Smad2 / 3 increased but without statistical significance (P>0.05); after treatment with Chinese medicine combined with lesion removal, the expression of MMP-9, VEGFA, and BMP2 decreased significantly. The expression of MMP-9, BMP-2, TGF-β1, and p-Smad2 / 3 increased significantly after treatment with positive drugs, while the expression of VEGFA and Smad2 / 3 increased but without statistical significance (P>0.05). The expression of MMP-9, VEGFA, TGF-β1, and Smad2 / 3 increased significantly after treatment with Chinese herbal medicine combined with positive drugs, while the expression of BMP-2 and p-Smad2 / 3 increased but without statistical significance (P>0.05). These results indicate that Chinese herbal medicine intervention promotes fibrosis and bone repair, as well as angiogenesis and tissue remodeling.

Claims

1. A Chinese medicine composition for treating osteomyelitis, characterized in that The invention is prepared from the following raw materials in proportion by weight: 15-18 parts of honeysuckle vine, 15-20 parts of patrinia scabra, 20-24 parts of raw astragalus, 8-12 parts of thorns of honeysuckle, 10-15 parts of cyathula, 10-15 parts of stir-fried atractylodes, 15-18 parts of poria, 10-15 parts of fructus aurantii, 8-12 parts of forsythia, 10-15 parts of angelica, 10-15 parts of ligusticum chuanxiong, 10-15 parts of stir-fried white peony root, 8-12 parts of yam and 25-30 parts of coix seed.

2. The Chinese medicine composition for treating osteomyelitis according to claim 1, wherein The invention is prepared from the following raw materials in proportion by weight: 15 parts of honeysuckle vine, 15 parts of patrinia scabra, 24 parts of raw astragalus, 10 parts of licorice thorn, 15 parts of cyathula, 10 parts of stir-fried atractylodes, 15 parts of poria, 10 parts of fructus aurantii, 10 parts of forsythia, 10 parts of angelica, 10 parts of ligusticum, 10 parts of stir-fried white peony root, 10 parts of yam and 25 parts of coix seed.

3. The Chinese medicine composition for treating osteomyelitis according to claim 1 or 2, wherein: The drug can be prepared into decoction, pills, tea, oral liquid, tablets, capsules, granules, paste or powder dosage forms.

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