Chronic wound healing and consolidation traditional Chinese medicine composition and application thereof

Abstract

The invention belongs to the technical field of medical nursing materials, and particularly relates to a chronic wound healing and consolidating traditional Chinese medicine composition and application thereof. The chronic wound healing and consolidating traditional Chinese medicine composition is prepared from the following components in parts by weight: 12 to 18 parts of radix salviae miltiorrhizae, 8 to 12 parts of radix notoginseng, 8 to 12 parts of vinegar-processed rhizoma corydalis, 12 to 18 parts of radix cyathulae, 8 to 12 parts of radix paeoniae rubra, 8 to 12 parts of flos carthami, 8 to 12 parts of pollen typhae, 8 to 12 parts of lignum sappan, 8 to 12 parts of spina gleditsiae and 3 to 7 parts of radix angelicae sinensis. The external ointment prepared from the traditional Chinese medicine composition has the effects of promoting blood circulation to remove blood stasis, relieving swelling and pain, astringing dampness and astringing sores and the like when applied to wound surfaces, remarkably accelerates the early healing process of the wound surfaces and effectively promotes growth of granulation tissues, so that the healing time is shortened, the economic cost of patients is saved, and pain caused by the wound surfaces can be effectively relieved; the comfort of the patient is improved. In addition, the raw materials of the medicine are from traditional Chinese medicines and have the characteristic of low toxicity, so that the treatment safety is ensured.

Description

Technical Field

[0001] The present invention belongs to the technical field of medical care products, and particularly relates to a traditional Chinese medicine combination for consolidating chronic wound healing and its application. Background Art

[0002] With the aging of the population, the increase in weight, and the increasing number of complications secondary to diseases such as decreased resistance, diabetes, and venous insufficiency, the number of patients with chronic wounds has increased significantly. According to statistics, about 1% of people may develop leg ulcers in their lifetime. In the United States alone, there are 3 - 6 million chronic wound patients every year, and the cost of treating these wounds is nearly $5 - 10 billion. In China, according to the results of epidemiological studies, the ratio of chronic wounds among surgical inpatients is about 1.5% to 20.3%. Chronic wounds are different from general traumatic wounds, and their injuries are more complex. They have different epidemiological and pathological characteristics in terms of pathogenesis, location, bacterial infection, pathological changes, etc., and the wound repair methods are also different. Their causes are mostly related to wound infection, poor local blood supply, diabetes, and radiation. Therefore, comprehensive treatment is required to fundamentally cure or control it within the normal range. Chronic wounds, also known as skin ulcers, refer to any wounds that fail to heal orderly within the expected time according to the normal wound healing process; wounds that have not healed for more than 3 months can become chronic wounds. Chronic wounds may exist for several years or even not heal for life, causing serious psychological and physiological pressure to patients, and thus bringing a huge economic burden to patients and the entire healthcare system. The treatment of chronic wounds involves multiple disciplines such as traditional Chinese medicine, hyperbaric oxygen, general surgery, orthopedics, dermatology, endocrinology, etc. The treatment methods are divided into non - surgical and surgical treatments. Non - surgical methods include: applying drugs to the wound; various dressings; various physical treatment means (sound, light, heat, electricity, etc.); negative pressure suction; closed negative pressure suction; growth factors; and hyperbaric oxygen treatment can also be used. Surgical methods include: debridement and suture; skin grafting; covering with large - sheet foreign skin; various local or axial skin flaps; microsurgical tissue transplantation; microsuture and transplantation of nerves, blood vessels, and tendons, etc.

[0003] Currently, although there are various drugs for treating wounds on the market, they all have problems such as poor curative effect and large side effects, with an unclear treatment effect, slow curative effect, easy recurrence; long treatment time cycle and heavy economic burden.

[0004] Therefore, there is a need to provide a traditional Chinese medicine combination for consolidating chronic wound healing. By scientifically extracting the effective components from traditional Chinese medicines and compounding various traditional Chinese medicines, it aims to promote chronic wound healing, save economic costs, relieve wound pain at the same time, and has no toxic side effects, providing a safe and effective treatment plan for patients. Summary of the Invention

[0005] The objective of the present invention is to provide a traditional Chinese medicine combination for consolidating chronic wound healing and its application, which is used to prepare an external ointment for application on the wound surface, having the effects of promoting blood circulation to remove blood stasis, detumescence and pain relief, astringing dampness and promoting granulation tissue growth, significantly accelerating the early healing process of the wound surface, effectively promoting the growth of granulation tissue, and thus shortening the healing time.

[0006] A traditional Chinese medicine combination for consolidating chronic wound healing, the raw materials for its preparation, by weight, include 12 - 18 parts of Salvia miltiorrhiza, 8 - 12 parts of Panax notoginseng, 8 - 12 parts of Corydalis yanhusuo processed with vinegar, 12 - 18 parts of Cyathula officinalis, 8 - 12 parts of Paeonia lactiflora, 8 - 12 parts of Carthamus tinctorius, 8 - 12 parts of Pollen Typhae, 8 - 12 parts of Sappanwood, 8 - 12 parts of Gleditsia sinensis spine, and 3 - 7 parts of Angelica sinensis.

[0007] The preparation method of the traditional Chinese medicine combination for consolidating chronic wound healing includes the following steps: preparing Salvia miltiorrhiza extract, Panax notoginseng extract, Corydalis yanhusuo extract processed with vinegar, Cyathula officinalis extract, Paeonia lactiflora extract, Carthamus tinctorius extract, Pollen Typhae extract, Sappanwood extract, Gleditsia sinensis spine extract, Angelica sinensis extract, and Astragalus membranaceus extract from Salvia miltiorrhiza, Panax notoginseng, Corydalis yanhusuo processed with vinegar, Cyathula officinalis, Paeonia lactiflora, Carthamus tinctorius, Pollen Typhae, Sappanwood, Gleditsia sinensis spine, Angelica sinensis, and Astragalus membranaceus respectively, pulverizing them and then stirring evenly to obtain the product.

[0008] Specific several traditional Chinese medicine components are selected and used in combination, taking advantage of the effective components of each component to jointly promote wound healing. Salvia miltiorrhiza is rich in components such as tanshinone and salvianolic acid, which have the effects of antioxidant, anti-inflammatory and improving microcirculation, and help to accelerate the formation of new blood vessels and tissue repair. The main active ingredient in Panax notoginseng is notoginsenoside, which has significant hemostatic and blood-activating effects, can reduce inflammatory reactions, promote blood circulation, and is beneficial to the rapid healing of wounds. Corydalis yanhusuo contains alkaloids such as tetrahydropalmatine, and these components can promote qi and relieve pain, promote blood circulation, relieve local pain and muscle tension. Cyathula officinalis contains a variety of glycoside compounds, which helps to activate blood circulation and dredge meridians, tonify the liver and kidneys, enhance the body's resistance, promote blood circulation, and thus support wound healing. Paeonia lactiflora contains components such as paeoniflorin, which endows it with the effects of cooling blood and detoxifying, activating blood circulation and removing stasis, can reduce local swelling, inhibit bacterial growth, and promote the absorption of inflammation. The main effective components of Carthamus tinctorius are safflower yellow pigment and safflower glycoside, which can dilate blood vessels, increase blood flow, further promote blood circulation, and help tissue repair. The flavonoids contained in Typha angustifolia have the effects of clearing heat and detoxifying, astringing and stopping bleeding, and are especially suitable for the treatment in case of bleeding; Sappanwood can achieve the effects of promoting blood circulation to heal wounds, reducing swelling and relieving pain through its specific chemical components, and promoting wound repair. Gleditsia sinensis contains components such as gleditsioside, which has a strong antibacterial and anti-inflammatory effect, protecting the wound from infection. The rich ferulic acid and angelica polysaccharide and other components in Angelica sinensis help to nourish blood and activate blood circulation, regulate the blood state, promote blood circulation, and ensure that the wound obtains sufficient nutrient supply. These traditional Chinese medicines have synergistic effects. Salvia miltiorrhiza and Panax notoginseng jointly optimize blood flow, ensuring both sufficient blood supply and controlling the bleeding risk; Paeonia lactiflora and Carthamus tinctorius used in combination enhance the ability of promoting blood circulation and removing stasis, promoting local blood circulation and metabolism; Cyathula officinalis combined with Angelica sinensis not only strengthens blood circulation, but also provides necessary nutritional support, which is beneficial to the repair of damaged tissues; Corydalis yanhusuo and Sappanwood combination promotes wound healing while relieving pain; and the combination of Typha angustifolia and Gleditsia sinensis not only has anti-inflammatory and bactericidal effects, but also has a certain effect of astringing and stopping bleeding. However, the effective components of the traditional Chinese medicine extracts on the market are less, contain a lot of impurities, and have poor efficacy. Therefore, it is necessary to make traditional Chinese medicine extracts by oneself, as much as possible to improve the effective components in the traditional Chinese medicine extracts, while reducing the impurity content, while improving the medicinal efficacy of the traditional Chinese medicine combination, reducing its side effects.

[0009] The preparation method of the said Salvia miltiorrhiza extract includes the following steps:

[0010] A1. After cleaning and drying and pulverizing Salvia miltiorrhiza to obtain Salvia miltiorrhiza powder, put it into an extraction kettle for supercritical carbon dioxide extraction to obtain Salvia miltiorrhiza extract and residue, and after rotary evaporation of the Salvia miltiorrhiza extract, obtain crude Salvia miltiorrhiza extract 1;

[0011] A2. Load the crude Salvia miltiorrhiza extract 1 onto a silica gel column, elute with eluent 1, collect the eluate, add ethyl acetate and let it stand for extraction. After the liquid surface is stratified, remove the aqueous phase. After rotary evaporation, the Salvia miltiorrhiza extract 1 is obtained.

[0012] A3. Mix the residue with water to obtain the Salvia miltiorrhiza fermentation substrate, perform fermentation treatment to obtain the fermentation broth, filter the fermentation broth to obtain the supernatant, and rotary evaporate and concentrate the supernatant at 40 - 50 °C to obtain the crude Salvia miltiorrhiza extract 2.

[0013] A4. Load the crude Salvia miltiorrhiza extract 2 onto a macroporous resin 1, elute with eluent 2, collect the eluate, and rotary evaporate and dry it at 40 - 50 °C to obtain the Salvia miltiorrhiza extract 2.

[0014] A5. Mix the Salvia miltiorrhiza extract 1 and the Salvia miltiorrhiza extract 2 to obtain the Salvia miltiorrhiza extract.

[0015] Both the liposoluble components (such as tanshinones) and water-soluble components (such as salvianolic acids) in Salvia miltiorrhiza have the effects of promoting blood circulation to remove blood stasis, reducing inflammation and swelling, etc. However, due to their different solubilities, when preparing the Salvia miltiorrhiza extract, the contents of the two are often unbalanced, resulting in the inability to exert their complete beneficial effects. Ordinary extraction contains more impurities, which not only fails to exert the medicinal value of the effective components but also has a large amount of impurities affecting the treatment effect. Through two processes of supercritical CO2 extraction and fermentation extraction, the liposoluble components (such as tanshinones) and water-soluble components (such as salvianolic acids) in Salvia miltiorrhiza are respectively extracted, giving full play to the multiple pharmacological effects of Salvia miltiorrhiza.

[0016] Preferably, in step A1, the drying temperature is 40 - 60 °C.

[0017] Preferably, the specific conditions for the supercritical carbon dioxide extraction are: pressure 23 - 28 MPa, entrainer flow rate 0.5 - 1.5 mL / min, extraction time 1.5 - 2.5 h, and extraction temperature 40 - 45 °C.

[0018] Preferably, the entrainer is an ethanol aqueous solution with a mass fraction of 92% - 96%.

[0019] Preferably, the silica gel column has a C18 alkyl group on its surface, a specific surface area of 350 - 400 m 2 / g, and a particle size of 15 - 25 μm.

[0020] In some preferred embodiments, the silica gel column is purchased from Sigma-Aldrich (Shanghai) Trading Co., Ltd.

[0021] Preferably, in step A2, the loading flow rate is 1 - 2 mL / min, and the elution flow rate is 2 - 4 mL / min.

[0022] Preferably, in the step A2, the eluent 1 is an acetonitrile aqueous solution with mass concentrations of 30%, 40%, 50%, 60%, and 70% respectively, and the addition amount of each acetonitrile aqueous solution with different mass fractions is the same.

[0023] Preferably, the addition amount of the eluent 1 is 70 - 80 times the mass of the crude Danshen extract 1.

[0024] First, an ethanol aqueous solution with a relatively high mass fraction is selected as the entrainer, which can improve the extraction ability of supercritical carbon dioxide for tanshinone. This may be because tanshinone has a certain polarity, and carbon dioxide is a non-polar solvent, which has a stronger solubility for non-polar or low-polarity solutes. Adding an entrainer can improve the extraction rate of tanshinone through the action of polar solvent molecules on solute molecules. The obtained crude Danshen extract 1 after extraction can be purified by silica gel column chromatography to further remove impurities and improve the purity of tanshinone in the Danshen extract 1.

[0025] Preferably, the mass ratio of the residue to water is 1:(10 - 20).

[0026] In the step A3, the specific steps of the fermentation treatment are as follows: Add Saccharomyces cerevisiae to the Danshen fermentation substrate, evacuate to complete vacuum and then introduce oxygen, ferment at 28 - 30 °C for 24 - 48 h, inoculate Lactobacillus plantarum, and ferment at 37 °C for 24 - 48 h under anaerobic conditions; inoculate Acetobacter xylinum and ferment at 28 - 30 °C for 24 - 48 h under aerobic conditions.

[0027] Preferably, the ventilation rate of oxygen when introducing oxygen is 0.1 - 0.5 vvm.

[0028] The Saccharomyces cerevisiae is active dry wine yeast with the strain number CICC 1892; the Lactobacillus plantarum has the strain number CICC 24936; the Acetobacter xylinum has the strain number ATCC 23767.

[0029] Preferably, the dosage of the Saccharomyces cerevisiae is 10 8 -10 9 CFU / g of the Danshen fermentation substrate; the dosage of the Lactobacillus plantarum is 10 7 -10 8 CFU / g of the Danshen fermentation substrate; the dosage of the Acetobacter xylinum is 10 7 -10 8 CFU / g of the Danshen fermentation substrate.

[0030] Preferably, the macroporous resin 1 is a weakly polar macroporous resin with an average pore diameter of 13 - 14 nm and a pore volume of 0.73 - 0.77 mL / g.

[0031] In some preferred embodiments, the macroporous resin 1 is purchased from Tianjin Haoju Resin Technology Co., Ltd., AB-8.

[0032] Preferably, in step A4, the loading flow rate is 1-3 mL / min, and the elution flow rate is 8-10 mL / min.

[0033] Preferably, in step A4, the eluent 2 is an aqueous ethanol solution with mass concentrations of 30%, 40%, 50%, 60%, and 70% respectively, and the addition amount of each ethanol aqueous solution with different mass fractions is the same.

[0034] Preferably, the addition amount of the eluent 2 is 70-80 times the mass of the crude Salvia extract 2.

[0035] The residue remaining after extraction is reused. Specific fermenting agents are used for distributed fermentation. According to the different fermentation conditions and functions of each fermenting agent, the residue is fermented in sequence. Saccharomyces cerevisiae grows rapidly under microaerobic conditions, secretes enzymes to decompose polysaccharides in the Salvia residue, and releases precursor substances of salvianolic acids. Lactobacillus plantarum produces organic acids through lactic acid fermentation, further destroying the cell structure of the Salvia residue and releasing salvianolic acids. Finally, Acetobacter xylinum produces acetic acid by oxidizing ethanol (yeast fermentation), further destroying the structure of the Salvia residue and promoting the release of salvianolic acids. The three fermenting agents act synergistically to efficiently extract salvianolic acid components from the Salvia residue. At the same time, according to the metabolic characteristics of microorganisms, the oxygen conditions and fermentation parameters are reasonably controlled to achieve the best extraction effect. Through macroporous resin chromatography purification, the purity and biological activity of salvianolic acid components are further improved. In addition, the temperature is controlled below 60°C throughout the process to avoid destroying heat-sensitive components in Salvia (such as tanshinone compounds). Finally, the tanshinone and salvianolic acid components are mixed to form a Salvia extract with multiple pharmacological effects, synergistically exerting its effect of promoting wound healing.

[0036] The preparation method of the Panax notoginseng extract includes the following steps:

[0037] B1. Enzymolysis: Wash and dry the Panax notoginseng, then crush it into powder. Adjust the temperature to 45-50°C and the pH to 4.5-5.5 by adding water. Add cellulase and pectinase, and after enzymolysis for 2-4 h, boil to inactivate the enzymes, and then concentrate and dry to obtain the enzymolyzed product of Panax notoginseng.

[0038] B2. Fermentation: Adjust the water content of the enzymolyzed product of Panax notoginseng to 30%-40% by adding water to obtain the fermentation substrate of Panax notoginseng. Add the Panax notoginseng fermenting agent for fermentation, filter, and rotary evaporate and concentrate at 50-60°C to obtain the fermented product of Panax notoginseng.

[0039] B3. Mix the Panax notoginseng fermented product with an ethanol aqueous solution with a mass concentration of 70% - 80%, reflux and extract at 80°C for 2 - 3 hours, repeat the extraction 2 - 3 times, filter the extract, combine the filtrates, and rotary evaporate and concentrate at 50 - 60°C to obtain the crude Panax notoginseng extract;

[0040] B4. Refer to step A4 to pass through the macroporous resin 2 to obtain the Panax notoginseng extract.

[0041] Preferably, the mass ratio of the Panax notoginseng powder to water is 1:(20 - 30).

[0042] Preferably, the addition amount of the cellulase is 80 - 120 U / g of Panax notoginseng powder; the addition amount of the pectinase is 50 - 100 U / g of Panax notoginseng powder.

[0043] Preferably, the Panax notoginseng fermenting agent includes one or more of Aspergillus niger, Rhizopus oryzae, Saccharomyces cerevisiae, Talaromyces flavus, Bacillus coagulans, Lactobacillus plantarum; Further preferably, it is Aspergillus niger, Rhizopus oryzae, Saccharomyces cerevisiae, Talaromyces flavus.

[0044] In some preferred embodiments, for the Aspergillus niger, the strain number is CICC 41254; for the Rhizopus oryzae, the strain number is CICC 3010; for the Saccharomyces cerevisiae, the strain number is CICC 1892; for the Talaromyces flavus, the strain number is CICC 41732.

[0045] Preferably, the dosage of the Aspergillus niger is 10 8 -10 9 CFU / g of Panax notoginseng fermentation substrate; the dosage of the Rhizopus oryzae is 10 7 -10 8 CFU / g of Panax notoginseng fermentation substrate; the dosage of the Saccharomyces cerevisiae is 10 7 -10 8 CFU / g of Panax notoginseng fermentation substrate; the dosage of the Talaromyces flavus is 10 7 -10 8 CFU / g of Panax notoginseng fermentation substrate.

[0046] In step B2, the specific steps for adding the Panax notoginseng fermenting agent for fermentation are: Add Aspergillus niger and Rhizopus oryzae to the Salvia miltiorrhiza fermentation substrate, ferment at 30 - 32°C for 24 - 48 hours under aerobic conditions, then inoculate Saccharomyces cerevisiae and Talaromyces flavus, and ferment at 28 - 30°C for 48 - 72 hours.

[0047] Preferably, the mass ratio of the Panax notoginseng fermented product to the ethanol aqueous solution with a mass concentration of 70% - 80% is 1:(10 - 20).

[0048] Preferably, in step B4, the macroporous resin 2 is a weakly polar macroporous resin with an average pore diameter of 9-10 nm and a pore volume of 0.75-0.95 mL / g.

[0049] In some preferred embodiments, the macroporous resin 2 is purchased from Tianjin Haoju Resin Technology Co., Ltd., DM130.

[0050] By first performing enzymatic hydrolysis, then fermentation, followed by alcohol extraction and separation and purification through macroporous resin, the content of active ingredients such as notoginsenosides in the notoginseng extract can be increased. The synergistic action of cellulase and pectinase can decompose components such as cellulose and pectin in the cell wall, effectively opening the cell wall barrier and releasing active ingredients such as notoginsenosides. Then, Aspergillus niger, Rhizopus oryzae, Saccharomyces cerevisiae, and Talaromyces flavus are introduced as fermenting agents, which can give full play to the synergistic effect of different microorganisms, significantly improving the extraction efficiency and pharmacological activity of notoginsenosides. The synergistic action of Aspergillus niger and Rhizopus oryzae can not only produce enzymes such as β-glucosidase that can hydrolyze the glycosyl group of notoginsenosides, promoting the transformation of notoginsenosides, but also destroy the cell wall through enzymatic hydrolysis, release active ingredients, and at the same time decompose complex polysaccharide substances into monosaccharides and oligosaccharides, providing easily utilizable substrates for Saccharomyces cerevisiae and Talaromyces flavus; Saccharomyces cerevisiae and Talaromyces flavus further transform and release active ingredients (such as notoginsenosides) in notoginseng through metabolic activities, and produce ethanol, organic acids, and secondary metabolites through metabolic activities, enhancing the antioxidant, anti-inflammatory, and angiogenesis-promoting pharmacological effects of the extract. The enzyme systems of the four fermenting agents are complementary, not only further increasing the content of active ingredients such as notoginsenosides in the notoginseng extract, but also improving its medicinal value, and can be applied in the pharmaceutical field.

[0051] The preparation method of the angelica extract includes the following steps:

[0052] C1. Wash and dry and crush angelica, add water to adjust the water content to 30% - 40% to obtain an angelica fermentation substrate, add an angelica fermenting agent, perform aerobic fermentation at 30 - 37 °C for 2 - 3 days, then perform anaerobic fermentation for 1 - 2 days, take out to obtain a fermentation broth, filter the fermentation broth to obtain a supernatant, and rotary evaporate and concentrate the supernatant at 50 - 60 °C to obtain an angelica fermentation product;

[0053] C2. Refer to step B3 to obtain a crude angelica extract;

[0054] C3. Refer to step B4 to pass through macroporous resin 3 to obtain an angelica extract.

[0055] The angelica fermenting agent includes one or more of Bacillus amyloliquefaciens, Bacillus subtilis, and Lactobacillus plantarum; further preferably, it is Bacillus amyloliquefaciens, Bacillus subtilis, and Lactobacillus plantarum.

[0056] In some preferred embodiments, the Bacillus amyloliquefaciens has a strain number of CICC 24519; the Bacillus subtilis has a strain number of CICC 10732; and the Lactobacillus plantarum has a strain number of CICC 24936.

[0057] Preferably, the dosage of Bacillus amyloliquefaciens is 10 8 -10 9 CFU / g of Angelica sinensis fermentation substrate; the dosage of Bacillus subtilis is 10 7 -10 8 CFU / g of Angelica sinensis fermentation substrate; the dosage of Lactobacillus plantarum is 10 8 -10 9 CFU / g of Angelica sinensis fermentation substrate.

[0058] Preferably, in step C4, the macroporous resin 3 is a non-polar macroporous resin with an average pore diameter of 8.5 - 9 nm and a pore volume of 1.35 - 1.65 mL / g.

[0059] In some preferred embodiments, the macroporous resin 3 is purchased from Tianjin Haoju Resin Technology Co., Ltd., HPD - 100.

[0060] Using Bacillus amyloliquefaciens, Bacillus subtilis, and Lactobacillus plantarum to ferment Angelica sinensis can significantly increase the contents of effective substances such as Angelica sinensis polysaccharide and ferulic acid in the Angelica sinensis extract, thereby improving the medicinal value of the final product. This may be because the three bacteria act synergistically. On the one hand, they can not only secrete cellulase and pectinase, and Lactobacillus plantarum produces organic acids, which jointly destroy the cell wall structure and release active ingredients, but also produce organic acids (such as lactic acid) to lower the environmental pH, further destroying the cell wall structure and releasing more effective substances such as Angelica sinensis polysaccharide and ferulic acid. On the other hand, under aerobic conditions, Bacillus amyloliquefaciens and Bacillus subtilis first decompose complex polysaccharides and proteins into small molecule substances such as monosaccharides and amino acids through enzymatic hydrolysis, providing easily utilizable substrates for the subsequent anaerobic fermentation of Lactobacillus plantarum. And Lactobacillus plantarum produces organic acids through lactic acid fermentation, which not only further promotes the transformation and release of active ingredients, but also organic acids and solvents such as ethanol can improve the solubility of ferulic acid and enhance its bioavailability.

[0061] The preparation methods of the processed Corydalis yanhusuo extract, Paeonia lactiflora extract, and Sappanwood extract are the same, which is the ethanol reflux extraction method.

[0062] Taking the processed Corydalis yanhusuo extract as an example, its preparation method includes the following steps: reflux extracting with an ethanol aqueous solution with a mass fraction of 70% for 2 - 3 times, each time for 1 - 2 h, combining the extraction solutions, concentrating and drying to obtain the tetrahydropalmatine extract.

[0063] The preparation methods of the Cyathula officinalis Kuan extract, Pollen Typhae extract, and Spina Gleditsiae extract are the same, which is the water extraction and alcohol precipitation method.

[0064] Taking the Cyathula officinalis Kuan extract as an example, its preparation method includes the following steps: decocting Cyathula officinalis Kuan with water for 2 - 3 times, each time for 1 - 2 hours, combining the decoction, concentrating, adding ethanol until the alcohol content reaches 70%, standing for precipitation, filtering, and drying to obtain the Achyranthes bidentata saponin extract.

[0065] The preparation method of the safflower extract is the water extraction method, including the following steps: decocting safflower with water for 2 - 3 times, each time for 1 - 2 hours, combining the decoction, concentrating, and drying to obtain the safflower extract.

[0066] The application of the traditional Chinese medicine combination for consolidating chronic wound healing is applied to the field of medicine. More preferably, it can be applied to the preparation of external ointments.

[0067] The raw materials for preparing the external ointment include the traditional Chinese medicine combination for consolidating chronic wound healing, yellow vaseline, and sesame oil.

[0068] Preferably, the mass ratio of the traditional Chinese medicine combination for consolidating chronic wound healing, yellow vaseline, and sesame oil is 1:(2.5 - 3.5):(0.8 - 1.2); more preferably, it is 1:3:1.

[0069] The yellow vaseline is of pharmaceutical grade, and the sesame oil is of food grade, both of which are commercially available.

[0070] The preparation method of the external ointment includes the following steps: grinding the traditional Chinese medicine combination for consolidating chronic wound healing to a particle size ≤ 2000 mesh. After heating the yellow vaseline to about 70°C and melting it, add the sesame oil and the external ointment, and stir evenly until it cools. Among them, small batches are made manually, and large batches are stirred with a mixer.

[0071] The application method of the external ointment includes the following steps: evenly applying the external ointment on the gauze or other dressings with a wooden spatula, and then applying it externally to the affected area.

[0072] Compared with the prior art, the advantages and beneficial effects of the present invention are as follows:

[0073] 1. The traditional Chinese medicine combination for consolidating chronic wound healing prepared by the present invention, by scientifically extracting the active ingredients from medicinal plants and preparing the traditional Chinese medicine combination into an external ointment, has the effects of promoting blood circulation to remove blood stasis, detumescence and pain relief, astringing dampness and promoting granulation, etc. It significantly accelerates the early healing process of the wound, effectively promotes the growth of granulation tissue, thereby shortening the healing time, saving the economic cost of patients, and can also effectively relieve the pain of the wound and improve the comfort of patients. In addition, the raw materials of this drug are traditional Chinese medicines, with the characteristics of low toxicity, ensuring the safety of treatment.

[0074] 2. The present invention selects specific traditional Chinese medicines as raw materials, and each raw material is compounded in a specific ratio. At the same time, each raw material is subjected to specific extraction according to the required active ingredients, maximizing the content of the active ingredients in the extracts of the traditional Chinese medicine combination. The active ingredients among the raw materials act synergistically to maximize the effects of promoting blood circulation to remove blood stasis, reducing swelling and alleviating pain, and promoting wound healing, and accelerating the repair and regeneration process of the wound to the greatest extent.

[0075] 3. Through two processes of supercritical CO2 extraction and fermentation extraction, the present invention extracts the liposoluble components (such as tanshinones) and water-soluble components (such as salvianolic acids) in Salvia miltiorrhiza respectively, giving full play to the multiple pharmacological effects of Salvia miltiorrhiza.

[0076] 4. By first enzymolysis, then fermentation, and separating and purifying through macroporous resin after alcohol extraction, the present invention can increase the content of active ingredients such as notoginsenosides in the notoginseng extract.

[0077] 5. The present invention selects Bacillus amyloliquefaciens, Bacillus subtilis, and Lactobacillus plantarum to ferment Angelica sinensis, which can significantly increase the content of active substances such as Angelica polysaccharide and ferulic acid in the Angelica sinensis extract, thereby improving the medicinal value of the final product. BRIEF DESCRIPTION OF THE DRAWINGS

[0078] In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required for the description of the embodiments or the prior art.

[0079] Figure 1 It is a photo of the wound surface of the patient before and after treatment with the external ointment described in Embodiment 1 of the present invention. a - Before treatment; b - After using the ointment for 8 days; c - After using the ointment for 12 days; d - After using the ointment for 18 days; e - After using the ointment for 24 days; f - After using the ointment for 27 days. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0080] The following will clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all of them. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the scope of protection of the present invention.

[0081] The raw materials used in the present invention are all commercially available. Specifically:

[0082] The surface of the silica gel column has a C18 alkyl group, with a specific surface area of 350 - 400 m 2 / g and a particle size of 15 - 25 μm, purchased from Sigma-Aldrich (Shanghai) Trading Co., Ltd.

[0083] Macroporous resin 1 is a weakly polar macroporous resin with an average pore diameter of 13 - 14 nm and a pore volume of 0.73 - 0.77 mL / g. It is purchased from Tianjin Haoju Resin Technology Co., Ltd., AB-8.

[0084] Macroporous resin 2 is a weakly polar macroporous resin with an average pore diameter of 9 - 10 nm and a pore volume of 0.75 - 0.95 mL / g. It is purchased from Tianjin Haoju Resin Technology Co., Ltd., DM130.

[0085] Macroporous resin 3 is a non-polar macroporous resin with an average pore diameter of 8.5 - 9 nm and a pore volume of 1.35 - 1.65 mL / g. It is purchased from Tianjin Haoju Resin Technology Co., Ltd., HPD-100.

[0086] Saccharomyces cerevisiae, strain number CICC 1892; Lactobacillus plantarum, strain number CICC 24936; Acetobacter xylinum, strain number ATCC 23767; Aspergillus niger, strain number CICC 41254; Rhizopus oryzae, strain number CICC 3010; Talaromyces flavus, strain number CICC 41732; Bacillus amyloliquefaciens, strain number CICC 24519; Bacillus subtilis, strain number CICC 10732.

[0087] Cellulase, enzyme activity about 11000 U / g; Pectinase, enzyme activity about 30000 U / g; Both are purchased from Ningxia Xiasheng Industrial Group Co., Ltd.

[0088] The yellow vaseline and sesame oil are both purchased from Shaanxi Panlong Yihai Pharmaceutical Co., Ltd.

[0089] Example 1

[0090] This example provides a traditional Chinese medicine combination for consolidating chronic wound healing. Its preparation raw materials, by weight, are 15 parts of Salvia miltiorrhiza, 10 parts of Panax notoginseng, 10 parts of Corydalis yanhusuo processed with vinegar, 15 parts of Cyathula officinalis, 10 parts of Paeonia lactiflora, 10 parts of Carthamus tinctorius, 10 parts of Typha angustata, 10 parts of Sappanwood, 10 parts of Gleditsia sinensis spine, and 5 parts of Angelica sinensis.

[0091] The preparation method of the traditional Chinese medicine combination for consolidating chronic wound healing includes the steps of: separately preparing Salvia miltiorrhiza extract, Panax notoginseng extract, Corydalis yanhusuo extract processed with vinegar, Cyathula officinalis extract, Paeonia lactiflora extract, Carthamus tinctorius extract, Typha angustata extract, Sappanwood extract, Gleditsia sinensis spine extract, Angelica sinensis extract, and Astragalus membranaceus extract from Salvia miltiorrhiza, Panax notoginseng, Corydalis yanhusuo processed with vinegar, Cyathula officinalis, Paeonia lactiflora, Carthamus tinctorius, Typha angustata, Sappanwood, Gleditsia sinensis spine, Angelica sinensis, and Astragalus membranaceus, and then pulverizing and stirring them evenly to obtain the product.

[0092] The preparation method of the Salvia miltiorrhiza extract includes the steps of:

[0093] A1. Wash the danshen, dry it, and crush it to obtain danshen powder. Put the danshen powder into an extraction kettle for supercritical carbon dioxide extraction to obtain danshen extract and residue. Rotate and evaporate the danshen extract to obtain crude danshen extract 1;

[0094] A2. Load crude danshen extract 1 onto a silica gel column and elute it with eluent 1. Collect the eluate, add ethyl acetate, let it stand for extraction, remove the aqueous phase after the liquid surface is stratified, and then rotate and evaporate to obtain danshen extract 1;

[0095] A3. Mix the residue with water to obtain a danshen fermentation substrate, conduct fermentation treatment to obtain a fermentation broth, filter the fermentation broth to obtain a supernatant, and rotate and concentrate the supernatant at 45°C to obtain crude danshen extract 2;

[0096] A4. Load crude danshen extract 2 onto macroporous resin 1 and elute it with eluent 2. Collect the eluate and rotate and dry it at 45°C to obtain danshen extract 2;

[0097] A5. Mix danshen extract 1 and danshen extract 2 to obtain the danshen extract.

[0098] In step A1, the drying temperature is 50°C.

[0099] The specific conditions for the supercritical carbon dioxide extraction are: pressure 25 MPa, entrainer flow rate 1 mL / min, extraction time 2 h, and extraction temperature 42°C.

[0100] The entrainer is an ethanol aqueous solution with a mass fraction of 95%.

[0101] In step A2, the loading flow rate is 1.5 mL / min and the elution flow rate is 3 mL / min.

[0102] In step A2, eluent 1 is an acetonitrile aqueous solution with mass concentrations of 30%, 40%, 50%, 60%, and 70% respectively, and the addition amount of each mass fraction of the acetonitrile aqueous solution is the same.

[0103] The addition amount of eluent 1 is 75 times the mass of crude danshen extract 1.

[0104] The mass ratio of the residue to water is 1:15.

[0105] In step A3, the specific steps of the fermentation treatment are: add Saccharomyces cerevisiae to the danshen fermentation substrate, pump it to complete vacuum and then introduce oxygen, ferment at 30°C for 24 h, inoculate Lactobacillus plantarum, and ferment at 37°C for 24 h under anaerobic conditions; inoculate Acetobacter xylinum and ferment at 30°C for 24 h under aerobic conditions.

[0106] When introducing oxygen, the oxygen ventilation volume is 0.3 vvm.

[0107] The dosage of the Saccharomyces cerevisiae is 10 9 CFU / g of the Salvia miltiorrhiza fermentation substrate; the dosage of the Lactobacillus plantarum is 10 7 CFU / g of the Salvia miltiorrhiza fermentation substrate; the dosage of the Acetobacter xylinum is 10 7 CFU / g of the Salvia miltiorrhiza fermentation substrate.

[0108] In the step A4, the loading flow rate is 2 mL / min, and the elution flow rate is 9 mL / min.

[0109] In the step A4, the eluent 2 is an aqueous ethanol solution with mass concentrations of 30%, 40%, 50%, 60%, and 70% respectively, and the addition amount of each ethanol aqueous solution with each mass fraction is the same.

[0110] The addition amount of the eluent 2 is 75 times the mass of the crude Salvia miltiorrhiza extract 2.

[0111] The preparation method of the Notoginseng extract comprises the following steps:

[0112] B1. Enzymolysis: Wash the Notoginseng, dry it, and pulverize it to obtain Notoginseng powder. Adjust the temperature to 48 °C and the pH to 5 by adding water. Add cellulase and pectinase, inactivate by boiling after enzymolysis for 3 h, concentrate and dry to obtain the Notoginseng enzymolysis product;

[0113] B2. Fermentation: Adjust the water content of the Notoginseng enzymolysis product to 35% by adding water to obtain the Notoginseng fermentation substrate, add the Notoginseng fermentation agent for fermentation, filter, and rotary evaporate and concentrate at 55 °C to obtain the Notoginseng fermentation product;

[0114] B3. Mix the Notoginseng fermentation product with an aqueous ethanol solution with a mass concentration of 75%, reflux and extract at 80 °C for 2.5 h, repeat the extraction 3 times, filter the extraction solution, combine the filtrates, and rotary evaporate and concentrate at 55 °C to obtain the crude Notoginseng extract;

[0115] B4. Refer to step A4 to pass through the macroporous resin 2 to obtain the Notoginseng extract.

[0116] The mass ratio of the Notoginseng powder to water is 1:25.

[0117] The addition amount of the cellulase is 100 U / g of the Notoginseng powder; the addition amount of the pectinase is 80 U / g of the Notoginseng powder.

[0118] The Notoginseng fermentation agent is Aspergillus niger, Rhizopus oryzae, Saccharomyces cerevisiae, and Talaromyces flavus.

[0119] The dosage of the Aspergillus niger is 10 8 CFU / g of the Notoginseng fermentation substrate; the dosage of the Rhizopus oryzae is 10 8CFU / g Panax notoginseng fermentation substrate; the dosage of Saccharomyces cerevisiae is 10 7 CFU / g Panax notoginseng fermentation substrate; the dosage of Talaromyces flavus is 10 8 CFU / g Panax notoginseng fermentation substrate.

[0120] In the step B2, the specific steps of adding the Panax notoginseng fermentation agent for fermentation are as follows: adding Aspergillus niger and Rhizopus oryzae to the Salvia miltiorrhiza fermentation substrate, fermenting at 32°C for 48 h under aerobic conditions, inoculating Saccharomyces cerevisiae and Talaromyces flavus, and fermenting at 30°C for 72 h.

[0121] The mass ratio of the Panax notoginseng fermented product to the ethanol aqueous solution with a mass concentration of 75% is 1:15.

[0122] The preparation method of the Angelica sinensis extract comprises the following steps:

[0123] C1. Cleaning, drying, and pulverizing Angelica sinensis, adjusting the water content to 35% by adding water to obtain the Angelica sinensis fermentation substrate, adding the Angelica sinensis fermentation agent, fermenting aerobically at 35°C for 3 days, then fermenting anaerobically for 1 day, taking out to obtain the fermentation broth, filtering the fermentation broth to obtain the supernatant, and rotary evaporating and concentrating the supernatant at 55°C to obtain the Angelica sinensis fermented product;

[0124] C2. Referring to step B3 to obtain the crude Angelica sinensis extract;

[0125] C3. Referring to step B4 to pass through macroporous resin 3 to obtain the Angelica sinensis extract.

[0126] The Angelica sinensis fermentation agent is Bacillus amyloliquefaciens, Bacillus subtilis, and Lactobacillus plantarum.

[0127] The dosage of Bacillus amyloliquefaciens is 10 9 CFU / g Angelica sinensis fermentation substrate; the dosage of Bacillus subtilis is 10 7 CFU / g Angelica sinensis fermentation substrate; the dosage of Lactobacillus plantarum is 10 8 CFU / g Angelica sinensis fermentation substrate.

[0128] The preparation methods of the Corydalis yanhusuo extract, Paeonia lactiflora extract, and Sappanwood extract are the same, which is the ethanol reflux extraction method.

[0129] Taking the Corydalis yanhusuo extract as an example, its preparation method comprises the following steps: reflux extracting with an ethanol aqueous solution with a mass fraction of 70% for 3 times, 2 h each time, combining the extraction solutions, concentrating, and drying to obtain the tetrahydropalmatine extract.

[0130] The preparation methods of the Cyathula officinalis extract, Pollen Typhae extract, and Spina Gleditsiae extract are the same, which is the water extraction and alcohol precipitation method.

[0131] Taking the extract of Cyathula officinalis Kuan as an example, its preparation method comprises the following steps: decocting Cyathula officinalis Kuan with water three times, each time for 2 hours, combining the decoctions, concentrating, adding ethanol until the ethanol content reaches 70%, standing for precipitation, filtering and drying to obtain the Achyranthes bidentata saponin extract.

[0132] The preparation method of the safflower extract is the water extraction method, and the steps are as follows: decocting safflower with water three times, each time for 2 hours, combining the decoctions, concentrating and drying to obtain the safflower extract.

[0133] The traditional Chinese medicine combination for consolidating chronic wound healing is used for preparing an external ointment.

[0134] The raw materials for preparing the external ointment are the traditional Chinese medicine combination for consolidating chronic wound healing, yellow petrolatum and sesame oil, and the mass ratio is 1:3:1.

[0135] The preparation method of the external ointment comprises the following steps: grinding the traditional Chinese medicine combination for consolidating chronic wound healing to a particle size of 2000 mesh, heating the yellow petrolatum to 70°C and melting it, then adding the sesame oil and the external ointment, and stirring evenly until it cools to obtain the product.

[0136] Example 2

[0137] The difference between this example and Example 1 is that for the traditional Chinese medicine combination for consolidating chronic wound healing, the raw materials, by weight, are 12 parts of Salvia miltiorrhiza, 12 parts of Panax notoginseng, 12 parts of Corydalis yanhusuo processed with vinegar, 15 parts of Cyathula officinalis Kuan, 10 parts of Paeonia lactiflora Pall., 10 parts of safflower, 10 parts of Pollen Typhae, 10 parts of Sappanwood, 10 parts of Gleditsia sinensis Lam., and 5 parts of Angelica sinensis.

[0138] Comparative Example 1

[0139] The difference between this comparative example and Example 1 is that for the traditional Chinese medicine combination for consolidating chronic wound healing, the raw materials, by weight, are 10 parts of Panax notoginseng, 10 parts of Corydalis yanhusuo processed with vinegar, 15 parts of Cyathula officinalis Kuan, 10 parts of Paeonia lactiflora Pall., 10 parts of safflower, 10 parts of Pollen Typhae, 10 parts of Sappanwood, 10 parts of Gleditsia sinensis Lam., and 5 parts of Angelica sinensis.

[0140] Comparative Example 2

[0141] The difference between this comparative example and Example 1 is that for the traditional Chinese medicine combination for consolidating chronic wound healing, the raw materials, by weight, are 15 parts of Salvia miltiorrhiza, 10 parts of Corydalis yanhusuo processed with vinegar, 15 parts of Cyathula officinalis Kuan, 10 parts of Paeonia lactiflora Pall., 10 parts of safflower, 10 parts of Pollen Typhae, 10 parts of Sappanwood, 10 parts of Gleditsia sinensis Lam., and 5 parts of Angelica sinensis.

[0142] Comparative Example 3

[0143] The difference between this comparative example and Example 1 is that the preparation method of the Salvia miltiorrhiza extract comprises the following steps:

[0144] A1. After cleaning and drying Salvia miltiorrhiza, it is crushed to obtain Salvia miltiorrhiza powder, which is put into an extraction kettle for supercritical carbon dioxide extraction to obtain Salvia miltiorrhiza extract and residue. After rotary evaporation of the Salvia miltiorrhiza extract, crude Salvia miltiorrhiza extract 1 is obtained;

[0145] A2. Load crude Salvia miltiorrhiza extract 1 onto a silica gel column and elute it with eluent 1. Collect the eluate, add ethyl acetate and let it stand for extraction. After the liquid surface is layered, remove the aqueous phase. After rotary evaporation, Salvia miltiorrhiza extract is obtained.

[0146] Comparative Example 4

[0147] The difference between this comparative example and Example 1 is that in step A3, the specific steps of the fermentation treatment are as follows: Saccharomyces cerevisiae is added to the Salvia miltiorrhiza fermentation substrate, pumped to complete vacuum and then oxygen is introduced. After fermentation at 30 °C for 24 h, Lactobacillus plantarum is inoculated and fermented at 37 °C for 24 h under anaerobic conditions.

[0148] Comparative Example 5

[0149] The difference between this comparative example and Example 1 is that the preparation method of the Panax notoginseng extract has the following steps:

[0150] B1. Fermentation: After cleaning and drying Panax notoginseng, it is crushed to obtain Panax notoginseng powder. The water content is adjusted to 35% by adding water to obtain the Panax notoginseng fermentation substrate, and a Panax notoginseng fermentation agent is added for fermentation. After filtration, it is rotary evaporated and concentrated at 55 °C to obtain the Panax notoginseng fermentation product;

[0151] B2. Mix the Panax notoginseng fermentation product with an ethanol aqueous solution with a mass concentration of 75%, reflux and extract at 80 °C for 2.5 h, repeat the extraction 3 times, filter the extract, combine the filtrates, and rotary evaporate and concentrate at 55 °C to obtain the crude Panax notoginseng extract;

[0152] B3. Refer to step A4 to pass through macroporous resin 2 to obtain Panax notoginseng extract.

[0153] Comparative Example 6

[0154] The difference between this comparative example and Example 1 is that the Panax notoginseng fermentation agent is Rhizopus oryzae, Saccharomyces cerevisiae, and Talaromyces flavus.

[0155] The dosage of Rhizopus oryzae is 10 9 CFU / g of Panax notoginseng fermentation substrate; the dosage of Saccharomyces cerevisiae is 10 8 CFU / g of Panax notoginseng fermentation substrate; the dosage of Talaromyces flavus is 10 8 CFU / g of Panax notoginseng fermentation substrate.

[0156] Comparative Example 7

[0157] The difference between this comparative example and Example 1 is that the Angelica sinensis fermentation agent is Bacillus subtilis and Lactobacillus plantarum.

[0158] The dosage of the Bacillus subtilis is 10 8 CFU / g of Angelica sinensis fermentation substrate; the dosage of the Lactobacillus plantarum is 10 9 CFU / g of Angelica sinensis fermentation substrate.

[0159] Performance Test

[0160] Mouse ear swelling experiment: Referring to the method in the paper "Study on the Anti-inflammatory Effect of Gentiana algida Pall." published by Yang Fang et al., the mouse ear swelling experiment was carried out. The external ointment was applied to the swollen area for a period of 24 days, and the dressing was changed 10 times in total. The inhibition rate of the external ointment on mouse ear swelling was measured. The higher the inhibition rate, the better the anti-inflammatory effect. The results are shown in Table 1.

[0161] Table 1 Measurement Results

[0162] Inhibition rate / % Example 1 89.74 Example 2 88.28 Control Example 1 82.95 Control Example 2 83.43 Control Example 3 85.86 Control Example 4 86.34 Control Example 5 84.40 Control Example 6 84.89 Control Example 7 85.86

[0163] According to statistics, the traditional Chinese medicine combination for consolidating chronic wound healing prepared in Examples 1-2 of the present invention has a relatively high inhibition rate on mouse ear swelling, indicating that its anti-inflammatory and anti-inflammatory effects are better. In Comparative Example 1, Salvia miltiorrhiza was not added; in Comparative Example 2, Panax notoginseng was not added; in Comparative Example 3, the water-soluble components of salvianolic acids were not collected; in Comparative Example 4, Acetobacter xylinum fermentation was not used; in Comparative Example 5, Panax notoginseng was not enzymolyzed first; in Comparative Example 6, Aspergillus niger was not added to the Panax notoginseng fermenting agent; in Comparative Example 7, Bacillus amyloliquefaciens was not added to the Angelica sinensis fermenting agent. The inhibition rate of the prepared traditional Chinese medicine combination is lower than that of Example 1 and Example 2. Therefore, the external ointment prepared in Example 1 of the present application was used for clinical trials subsequently.

[0164] Data collection of clinical efficacy evaluation indicators:

[0165] 1. Wound pain

[0166] The Numerical Rating Scale (NRS) was used. When removing the inner dressing during dressing change and during wound treatment, the Numerical Rating Scale (NRS) was used to evaluate the pain degree of the patient. The scoring range is 0-10 points, and the higher the score, the more severe the pain. It was evaluated before and after each treatment.

[0167] 2. Granulation color

[0168] Granulation color scoring: Before treatment and during each dressing change, the score was completed after comparing with the granulation tissue color comparison card. Cure: 0 points; Ruddy: 1 point; Dark red: 2 points; Grayish white: 3 points; Purple black: 4 points.

[0169] 3. Wound area

[0170] Before treatment and during each dressing change, the ulcer area was measured. Wound area (cm2) = longest diameter of the wound (cm) × widest diameter of the wound (cm).

[0171] 4. Judgment Criteria

[0172] It is formulated with reference to the "Diagnostic and Therapeutic Criteria for Traditional Chinese Medicine Syndromes and Diseases" of the State Administration of Traditional Chinese Medicine. Clinical cure: The wound surface is completely healed and the symptoms disappear. Marked effect: The wound healing rate is ≥ 75%, the granulation tissue is light red, there is no pus, the obvious swelling around the wound disappears significantly, and the pain is basically relieved. Effective: 25% ≤ wound healing rate < 75%, the granulation tissue is bright red, there may be a small amount of pus, and the wound circumference is slightly swollen. Ineffective: The wound healing rate < 25% or there is no obvious change or even aggravation. The total effective rate = (number of cured cases + number of markedly effective cases + number of effective cases) / total number of cases × 100%.

[0173] 5. Data Statistical Results:

[0174] From January 2024 to December 2024, 20 patients who visited the combined wound care and traditional Chinese medicine outpatient clinic were collected as the research objects and randomly divided into 2 groups: a control group and a treatment group. Inclusion criteria: age < 80 years old; wound surface area ≤ 20 cm 2 ; conservative dressing change treatment is feasible; unable to undergo surgical treatment for other reasons; patients with unhealed chronic lower extremity venous ulcer wounds; exclusion criteria: previous history of surgery for lower extremity venous ulcer disease; severe systemic infection; withdrawal or transfer to another hospital for treatment midway. Divided into the control group (8 males and 2 females, average age 67 ± 12.5 years old) by digital lottery; treatment group (7 males and 3 females, average age 65 ± 13.8 years old); the control group implemented moist healing + new dressing change, and the treatment group used the traditional Chinese medicine paste prepared in Example 1 for external application treatment. The wound healing conditions of the two groups were analyzed by SPSS 26.0, and the data statistics are shown in Tables 2 to 5.

[0175] Table 2 Comparison of wound pain scores of two groups of patients before and after treatment [mean ± standard deviation, points]

[0176] Group Number of cases Before treatment 3 days after treatment 5 days after treatment Control group 10 4.2±1.2 3.3±1.0 2.5±0.74 Treatment group 10 4.3±1.1 2.9±0.88 2.1±0.62

[0177] Table 3 Comparison of granulation tissue color scores of two groups of patients before and after treatment [mean ± standard deviation, points]

[0178] Group Number of cases Before treatment 3 days after treatment 5 days after treatment Control group 10 3.3±1.1 2.5±0.73 1.9±0.57 Treatment group 10 3.2±1.3 2.3±0.69 1.6±0.46

[0179] Table 4 Comparison of wound areas of two groups of patients before and after treatment [mean ± standard deviation, points]

[0180] Group Number of cases Before treatment 5 days after treatment Control group 10 11.5±3.5 6.32±1.96 Treatment group 10 11.2±3.3 4.29±1.24

[0181] Table 5 Comparison of wound healing effective rates of two groups of patients before and after treatment [n, (%)]

[0182] Group Number of cases Cured Markedly effective Effective Invalid Effective rate (%) Control group 10 0 3 4 2 70％ Treatment group 10 1 4 4 1 90％

[0183] As can be seen from Tables 2 to 5, compared with ordinary dressings, the paste prepared in Example 1 is applied externally, which can promote the dissolution of ulcer necrotic tissue, facilitate the growth of granulation tissue, accelerate wound healing, and relieve local pain of patients.

[0184] Practical case: A 78-year-old male had chronic lower limb venous ulcers for more than 20 months, with repeated ulcers and infections. The external paste prepared in Example 1 was used for 27 days, and the dressing was changed 10 times in total. Photos of the wound before and after treatment are shown in Figure 1 . It can be seen from Figure 1 that the wound has improved significantly and healed.

[0185] The above is the preferred embodiment of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and modifications can be made, and these improvements and modifications should also be regarded as the protection scope of the present invention.

Claims

1. A traditional Chinese medicine combination for consolidating chronic wound healing, characterized in that, Its preparation raw materials, by weight, include 12 - 18 parts of Salvia miltiorrhiza, 8 - 12 parts of Panax notoginseng, 8 - 12 parts of Corydalis yanhusuo processed with vinegar, 12 - 18 parts of Cyathula officinalis, 8 - 12 parts of Paeonia lactiflora, 8 - 12 parts of Carthamus tinctorius, 8 - 12 parts of Pollen typhae, 8 - 12 parts of Sappanwood, 8 - 12 parts of Spina Gleditsiae, and 3 - 7 parts of Angelica sinensis.

2. A preparation method of the traditional Chinese medicine combination for consolidating chronic wound healing according to claim 1, characterized in that, It includes the following steps: Separate Salvia miltiorrhiza, Panax notoginseng, Corydalis yanhusuo processed with vinegar, Cyathula officinalis, Paeonia lactiflora, Carthamus tinctorius, Pollen typhae, Sappanwood, Spina Gleditsiae, Angelica sinensis, and Astragalus membranaceus into Salvia miltiorrhiza extract, Panax notoginseng extract, Corydalis yanhusuo extract, Cyathula officinalis extract, Paeonia lactiflora extract, Carthamus tinctorius extract, Pollen typhae extract, Sappanwood extract, Spina Gleditsiae extract, Angelica sinensis extract, and Astragalus membranaceus extract respectively. After pulverizing them, stir evenly to obtain the product.

3. The preparation method of the traditional Chinese medicine combination for consolidating chronic wound healing according to claim 2, characterized in that, The preparation method of the Salvia miltiorrhiza extract includes the following steps: A1. Wash and dry - pulverize Salvia miltiorrhiza to obtain Salvia miltiorrhiza powder, put it into an extraction kettle for supercritical carbon dioxide extraction to obtain Salvia miltiorrhiza extract and residue. Rotate - evaporate the Salvia miltiorrhiza extract to obtain crude Salvia miltiorrhiza extract 1; A2. Load crude Salvia miltiorrhiza extract 1 onto a silica gel column and elute it with eluent 1. Collect the eluate, add ethyl acetate and let it stand for extraction. After the liquid surface is stratified, remove the aqueous phase, and then perform rotary evaporation to obtain Salvia miltiorrhiza extract 1; A3. Mix the residue with water to obtain a Salvia miltiorrhiza fermentation substrate, perform fermentation treatment to obtain a fermentation broth, filter the fermentation broth to obtain a supernatant, and rotary - evaporate and concentrate the supernatant at 40 - 50 °C to obtain crude Salvia miltiorrhiza extract 2; A4. Load crude Salvia miltiorrhiza extract 2 onto macroporous resin 1 and elute it with eluent 2. Collect the eluate and perform rotary - evaporation and drying at 40 - 50 °C to obtain Salvia miltiorrhiza extract 2; A5. Mix Salvia miltiorrhiza extract 1 and Salvia miltiorrhiza extract 2 to obtain the Salvia miltiorrhiza extract.

4. The preparation method of the traditional Chinese medicine combination for consolidating chronic wound healing according to claim 3, characterized in that, In step A3, the specific steps of the fermentation treatment are as follows: Add Saccharomyces cerevisiae to the Salvia miltiorrhiza fermentation substrate, pump it to complete vacuum and then introduce oxygen, ferment at 28 - 30 °C for 24 - 48 h, then inoculate Lactobacillus plantarum and ferment at 37 °C for 24 - 48 h under anaerobic conditions; inoculate Acetobacter xylinum and ferment at 28 - 30 °C for 24 - 48 h under aerobic conditions.

5. The preparation method of the traditional Chinese medicine combination for consolidating chronic wound healing according to claim 4, characterized in that, The dosage of the Saccharomyces cerevisiae is 10 8 -10 9 CFU / g of the Salvia miltiorrhiza fermentation substrate; the dosage of the Lactobacillus plantarum is 10 7 -10 8 CFU / g of the Salvia miltiorrhiza fermentation substrate; the dosage of the Acetobacter xylinum is 10 7 -10 8 CFU / g of the Salvia miltiorrhiza fermentation substrate.

6. The preparation method of the traditional Chinese medicine combination for consolidating chronic wound healing according to claim 3, characterized in that, The preparation method of the Panax notoginseng extract includes the following steps: B1. Enzymatic hydrolysis: Wash and dry - pulverize Panax notoginseng to obtain Panax notoginseng powder, adjust the temperature to 45 - 50 °C and pH to 4.5 - 5.5 with water, add cellulase and pectinase, perform enzymatic hydrolysis for 2 - 4 h, then boil to inactivate the enzymes, and concentrate and dry to obtain Panax notoginseng enzymolysis product; B2. Fermentation: Adjust the water content of the Panax notoginseng enzymolysis product to 30% - 40% with water to obtain a Panax notoginseng fermentation substrate, add a Panax notoginseng fermentation agent for fermentation, filter, and rotary - evaporate and concentrate at 50 - 60 °C to obtain a Panax notoginseng fermented product; B3. Mix the Panax notoginseng fermented product with an ethanol aqueous solution with a mass concentration of 70% - 80%, reflux and extract at 80 °C for 2 - 3 h, repeat the extraction 2 - 3 times, filter the extraction solution, combine the filtrates, and rotary - evaporate and concentrate at 50 - 60 °C to obtain crude Panax notoginseng extract; B4. Refer to step A4 to pass through macroporous resin 2 to obtain Panax notoginseng extract.

7. The preparation method of the traditional Chinese medicine combination for consolidating chronic wound healing according to claim 6, characterized in that, The addition amount of the cellulase is 80 - 120 U / g of pseudo-ginseng powder; the addition amount of the pectinase is 50 - 100 U / g of pseudo-ginseng powder.

8. The preparation method of the traditional Chinese medicine combination for consolidating chronic wound healing according to claim 7, characterized in that, The pseudo-ginseng fermenting agent includes one or more of Aspergillus niger, Rhizopus oryzae, Saccharomyces cerevisiae, Talaromyces flavus, Bacillus coagulans, Lactobacillus plantarum.

9. A traditional Chinese medicine combination for consolidating chronic wound healing according to claim 1 is used for preparing an external ointment.

10. Use of the traditional Chinese medicine combination for consolidating chronic wound healing according to claim 9, characterized in that, The application method of the external ointment includes the following steps: scraping the external ointment on a gauze or other dressing, and then applying it externally on the affected area.

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