Huo xue san gelatinous plaster and its preparation method and application

Abstract

This invention provides a blood-activating powder gel patch, its preparation method, and its application. The gel patch consists of lyophilized blood-activating powder and a gel matrix. The lyophilized blood-activating powder is obtained by extracting and freeze-drying seven herbs: *Artemisia capillaris*, *Kochia scoparia*, *Phellodendron chinense*, *Scutellaria barbata*, *Eupolyphaga sinensis*, *Polygonum cuspidatum*, and raw *Arisaema heterophyllum*. The gel matrix contains a cross-linking polymer NP-700, a thickener polyvinylpyrrolidone K90, a cross-linking agent aluminum hydroxide, a cross-linking regulator tartaric acid, and a humectant glycerin. In preparation, the cross-linking agent and cross-linking polymer are added to glycerin and mixed to obtain phase A. The thickener and cross-linking regulator are dissolved in purified water and left overnight to obtain phase B. The lyophilized blood-activating powder is dissolved in ethanol to obtain phase C. Phase C is mixed with phase B and then with phase A and stirred evenly before being applied. This invention transforms traditional blood-activating powder into a gel patch, which is stable in shape, has good adhesion, can be repeatedly applied and removed without staining clothing, and simultaneously improves transdermal penetration and adhesion. It also overcomes the irritation of raw *Arisaema heterophyllum*, achieving safe external use. Pharmacological studies have shown that this plaster has a significant therapeutic effect on soft tissue injuries and osteoarthritis.

Description

Technical Field

[0001] The present invention belongs to the technical field of pharmaceutical preparations, specifically relates to the technical field of traditional Chinese medicine external plaster, and particularly relates to a Huoxuesan gel plaster, its preparation method and application in the preparation of drugs for treating traumatic injuries. Background Art

[0002] Fracture is a common clinical traumatic disease. The interruption of the integrity or continuity of bones can occur at all ages, especially in children and the elderly. Patients often show symptoms such as local pain, tenderness, swelling, and ecchymosis. Analyzed from the pathological mechanism, the fracture ends often lead to damage of the surrounding soft tissues, affecting the arteries and veins that supply local blood, and then triggering a series of pathological changes such as hypercoagulable state of blood, ischemia, and vascular endothelial dysfunction. Blood stasis is the core symptom characteristic, which often accompanies fractures and affects the healing process.

[0003] Traditional Chinese medicine clinical treatment of fractures focuses on promoting blood circulation to remove blood stasis, starting from both internal causes and external symptoms. Huoxuesan is a commonly used traditional Chinese medicine compound preparation in clinical practice, which consists of seven herbs: Artemisia anomala S. Moore, Kochia scoparia (L.) Schrad., Phellodendron amurense Rupr., Scutellaria barbata D. Don, Eupolyphaga sinensis Walker, Polygonum cuspidatum Sieb. & Zucc., and Arisaema heterophyllum Blume. The whole formula has the effects of clearing heat and promoting diuresis, breaking blood and removing blood stasis, and can promote blood circulation to remove blood stasis, reduce swelling and relieve pain. It is mainly used for treating traumatic injuries such as fractures, fissured fractures, and soft tissue contusions, with definite curative effects and is widely used in traditional Chinese medicine orthopedics.

[0004] At present, the related research on Huoxuesan is relatively weak. The traditional plaster dosage form of Huoxuesan is mostly to stir and mix the traditional Chinese medicine compound powder with maltose to form a thick paste, and directly coat it on the non-woven fabric and apply it to the affected area. There are many deficiencies in the actual application of this simple dosage form: the adhesion performance of the paste is poor, it is easy to fall off from the skin, it is inconvenient to use and easy to pollute clothes. At the same time, the water content and drug loading amount of the paste are difficult to control, affecting the stable exertion of drug efficacy. Therefore, it is of great clinical significance and application value to improve and deeply study the plaster dosage form of Huoxuesan and develop a new dosage form that is convenient to use, has good adhesion, does not pollute clothes, and has stable drug efficacy. [[ID=​​​​​Technical problem to be solved: In view of the above-mentioned technical problems, the present invention provides a topical Chinese medicine wash that can improve acne and seborrheic dermatitis, as well as its preparation method and application.

[0007] Technical solution: A blood-activating powder gel plaster, composed of blood-activating powder freeze-dried powder and a gel matrix; the blood-activating powder freeze-dried powder includes seven herbs: Artemisia annua, Kochia scoparia, Phellodendron chinense, Scutellaria barbata, Eupolyphaga sinensis, Polygonum cuspidatum, and Arisaema heterophyllum; the gel matrix contains a cross-linked polymer, a thickener, a cross-linking agent, a cross-linking regulator, and a moisturizer.

[0008] The aforementioned crosslinked polymer is a partially neutralized polyacrylic acid compound. The partially neutralized compound is at least one of NP-600 with a neutralization degree of 70%, NP-700 with a neutralization degree of 50%, and NP-800 with a neutralization degree of 35%. The tackifier is at least one of sodium carboxymethyl cellulose, sodium methyl cellulose, polyvinyl alcohol, polyvinylpyrrolidone K90, gelatin, and gum arabic. The crosslinking agent is at least one of aluminum hydroxyl, aluminum hydroxide, aluminum chloride, aluminum sulfate, and alum. The crosslinking regulator is at least one of tartaric acid, citric acid, lactic acid, EDTA, or EDTA-2Na. The humectant is at least one of propylene glycol, glycerin, 1,3-butanediol, and polyethylene glycol.

[0009] The weight parts of each component in the above gel matrix are as follows: 4.5 parts of cross-linking polymer, 8.8 parts of thickener, 0.75 parts of cross-linking agent, 1 part of cross-linking regulator, 20 parts of humectant, and 20 parts of purified water; the weight parts of the blood-activating powder lyophilized powder are 2 parts.

[0010] The weight ratio of the above-mentioned Artemisia annua, Kochia scoparia, Scutellaria barbata, Polygonum cuspidatum, Arisaema heterophyllum, Phellodendron chinense, and Eupolyphaga sinensis is 170:170:160:170:170:85:75.

[0011] The above-mentioned blood-activating powder freeze-dried powder was prepared by the following method: taking seven herbs, namely, Artemisia capillaris, Kochia scoparia, Phellodendron chinense, Scutellaria barbata, Eupolyphaga sinensis, Polygonum cuspidatum, and Arisaema heterophyllum, and using 75% ethanol as solvent, refluxed twice with a volume of 14 times, each time for 30 minutes, combining the extracts, concentrating, and freeze-drying to obtain the powder.

[0012] The method for preparing Huoxue San gel plaster includes the following steps: (1) Add cross-linking agent and cross-linking polymer to moisturizer and mix evenly to obtain phase A; (2) Dissolve thickener and cross-linking regulator in purified water and let stand overnight to obtain phase B; (3) Dissolve Huoxue San freeze-dried powder in ethanol to obtain phase C; (4) Add phase C to phase B and mix evenly to obtain phase D; then add phase D to phase A, mix evenly, and apply to obtain Huoxue San gel plaster.

[0013] The mixing conditions in step (1) are room temperature and stirring at 200 rpm for 3 min; the mixing conditions in step (4) are room temperature and stirring at 200 rpm for 3 min.

[0014] The ethanol mentioned in step (3) is 75% ethanol, and the mass-volume ratio of Huoxue San freeze-dried powder to ethanol is 1g:1mL.

[0015] The above-mentioned blood-activating powder gel plaster is used in the preparation of medicines for treating bruises and sprains.

[0016] The above-mentioned sprains and bruises are either soft tissue injuries or osteoarthritis.

[0017] Beneficial Effects: This invention successfully transforms the traditional blood-activating powder, which is a simple dosage form mixed with maltose, into a gel patch. Traditional powders require on-the-spot preparation, have poor adhesion, and are prone to falling off and staining clothing, resulting in low patient compliance. In contrast, the blood-activating powder gel patch prepared by this invention has stable molding, an intact appearance, and suitable adhesive properties. It can be repeatedly applied and removed without staining clothing, significantly improving the convenience of clinical use. More importantly, while maintaining good adhesion, the transdermal penetration rate of this invention's gel patch is not negatively affected; on the contrary, it achieves a simultaneous improvement in both adhesion and transdermal penetration rate. This effect is difficult to predict in formulation, as increasing matrix adhesion usually hinders drug release. However, this invention overcomes this technical challenge by optimizing the matrix composition and ratio.

[0018] It is worth noting that the Huoxue San formula contains raw Arisaema heterophyllum, which has a certain degree of irritation. Traditionally, it is often processed to reduce its toxicity. However, in this invention, Huoxue San is extracted, freeze-dried, and then combined with a gel matrix. The resulting gel patch was continuously administered in pharmacodynamic experiments on soft tissue injury and osteoarthritis in rats. No skin irritation was observed, and histopathological examination showed no obvious inflammatory cell infiltration or tissue damage. This result indicates that the gel patch matrix used in this invention can effectively encapsulate the active ingredients in raw Arisaema heterophyllum, and may also provide a certain degree of sustained release and barrier effect on irritating components, achieving safe external application of raw Arisaema heterophyllum and overcoming the limitations of its traditional applications.

[0019] From a pharmacodynamic perspective, the gel patch of this invention exhibits good therapeutic effects in a soft tissue injury model. From day 3 of administration, the swelling degree in the patch group was significantly lower than that in the model group, comparable to the efficacy of the powder group; by day 7, the swelling degree in the patch group decreased to near the level of the control group, and the appearance score of soft tissue injury was significantly improved. In an osteoarthritis model, the mechanical pain threshold and gait parameters in the patch group were significantly better than those in the model group, the pathological changes in the knee joint synovium were significantly reduced, and the levels of serum inflammatory factors such as TNF-α, IL-1β, IL-6, and PGE2 were significantly reduced. These pharmacodynamic data confirm that the gel patch of this invention not only retains the efficacy of traditional powders but also shows a superior trend in certain indicators. This maintenance and improvement of efficacy after dosage form conversion has unexpected technical effects.

[0020] Furthermore, the preparation process of the gel patch of this invention is simple and controllable. It employs a method of preparing phases A, B, and C separately and then mixing them. The process conditions are mild, requiring no high-temperature treatment, which helps maintain the stability of the traditional Chinese medicine components. The formula contains 32% glycerin by weight, which acts as both a moisturizer to maintain the patch's moisture content and performance, and also serves as a preservative, eliminating the need for additional preservatives and simplifying the formula composition. The entire preparation process does not use large amounts of organic solvents, has no explosion-proof requirements, low production costs, and is environmentally friendly, demonstrating good prospects for industrialization. Attached Figure Description

[0021] Figure 1 Comparison of inflammatory factors in the leg muscles of rats in different groups ( (±s, n=6) Note: Compared with the normal control group at the same time point, ★P<0.05, ★★P<0.01; compared with the model group at the same time point, #P<0.05, ##P<0.01. K: blank group, M: model group, Y: positive drug control group, S: powder group, T: plaster group, same as the figure below.

[0022] Figure 2 Local histopathological results of rats in each group (40×).

[0023] Figure 3 Pathological changes in the knee joints of rats in each group (20×).

[0024] Figure 4 Comparison of inflammatory factors in the knee joints of rats in different groups ( ±s, n=6).

[0025] Figure 5 Images of the right hind limb of rats in each group at days 1, 3, 5, and 7. Detailed Implementation

[0026] The present invention will be further described below with reference to specific embodiments. It should be noted that the following embodiments are exemplary and are not intended to limit the scope of the invention. After reading the contents of this invention, those skilled in the art can make various modifications or alterations to the invention, and these equivalent forms also fall within the scope defined by the claims herein.

[0027] 1. Materials and Reagents

[0028] Artemisia annua (batch number 220512, purchased from Jiangsu Ninghe Pharmaceutical Co., Ltd.), Kochia scoparia (batch number 220901, purchased from Bozhou Jingwan Traditional Chinese Medicine Pieces Factory), Scutellaria barbata (batch number 221001, purchased from Anhui Huchuntang Traditional Chinese Medicine Pieces Co., Ltd.), Polygonum cuspidatum (batch number A221224, purchased from Bozhou Yonggang Traditional Chinese Medicine Pieces Factory Co., Ltd.), and Arisaema heterophyllum (batch number D2112020, purchased from Sichuan Xinhehua Traditional Chinese Medicine Pieces Co., Ltd.) Phellodendron bark (batch number 220801, purchased from Chongqing Wanli Pharmaceutical Co., Ltd.), ground beetle (batch number 220921, purchased from Suzhou Boyuan Pharmaceutical Co., Ltd.); demethylene berberine (batch number: PS020675, purity: ≥98.0%), palmatine hydrochloride (batch number: PS012545, purity: ≥98.5%), vanillic acid (batch number: PS010559, purity: ≥98.0%), luteolin ( The following reference standards were purchased from Chengdu Pusi Biotechnology Co., Ltd.: Batch No.: PU0033-0025, purity: ≥99.0%; Baicalein (Batch No.: PS011541, purity: ≥98.0%); Apigenin (Batch No.: PS011063, purity: ≥99.0%); Formic acid (Batch No.: PS011063, chromatographic grade, Shanghai Maclean Biochemical Technology Co., Ltd.); Acetonitrile (Batch No.: PS011063, chromatographic grade, Shanghai Maclean Biochemical Technology Co., Ltd.); Ethanol (Batch No.: PS011063, analytical grade, Shanghai Titan Technology Co., Ltd.); Purified water (laboratory preparation); NP-700 (Jiangxi Alpha High-Tech Pharmaceutical Co., Ltd.); Aluminum hydroxyl (Jiangxi Alpha High-Tech Pharmaceutical Co., Ltd.); Tartaric acid (Chengdu Huayi Pharmaceutical Excipients Manufacturing Co., Ltd.); Polyvinylpyrrolidone K90 (PVP) K90, Shanghai Senjun Chemical Preparations and Excipients Co., Ltd.; Glycerin (batch number: 2151038811, FPG Oleochemicals Sdn. Bhd.).

[0029] 2. Instruments and Equipment

[0030] RE-52AA rotary evaporator (Shanghai Lichen Instrument Technology Co., Ltd.); electronic analytical balance (Mettler-Toledo, Switzerland); SCIENTZ-10N freeze dryer (Ningxinzhi Biotechnology Co., Ltd.); HPLC chromatograph (Waters Technology (Shanghai) Co., Ltd.); pulverizer (Zhejiang Hongjingtian Industry & Trade Co., Ltd.); ultrasonic cleaner (Kunshan Sheng Instrument Co., Ltd.); circulating water multi-purpose vacuum pump (Shanghai Qiangqiang Industrial Development Co., Ltd.); digital force gauge; digital force gauge test stand; initial viscosity tester; universal mixer (Shanghai Weiguang Machinery Manufacturing Co., Ltd.); high-speed refrigerated centrifuge (Hunan Kecheng Instrument Equipment Co., Ltd.).

[0031] 3. Laboratory animals

[0032] Thirty 8-week-old male SPF-grade SD rats, weighing approximately 200 g, were purchased from Jiangsu Qinglongshan Biotechnology Co., Ltd. [Experimental Animal Production License No. SCXK (Su) 2024-0001] and housed at the Experimental Animal Center of Nanjing University of Chinese Medicine. The indoor temperature was 20–25℃, relative humidity 45%–70%, with 12-hour light / dark cycles. Rats had free access to food and water and underwent acclimatization feeding for one week. The experiment was approved by the Ethics Committee of the Experimental Animal Center of Nanjing University of Chinese Medicine (Ethics No. 202503A028).

[0033] 4. Experimental methods and results

[0034] See the following examples for details.

[0035] Example 1: Preparation of Blood-Activating Powder

[0036] Weigh out the following herbs: Artemisia capillaris 170 g, Kochia scoparia 170 g, Scutellaria barbata 160 g, Polygonum cuspidatum 170 g, Arisaema heterophyllum 170 g, Phellodendron chinense 85 g, and Eupolyphaga sinensis 75 g. Mix the above seven herbs, except for Arisaema heterophyllum 170 g, dry at 60 ℃, pulverize and pass through a 120-mesh sieve. Pulverize Arisaema heterophyllum 170 g separately and pass through a 120-mesh sieve. Mix the two powders by adding them in equal amounts, mix well and pass through a 120-mesh sieve to obtain the final product.

[0037] Example 2: Study on the reflux extraction process of Huoxue San (single-factor investigation)

[0038] 10.0 g of Huoxue San powder was weighed and extracted by reflux. The appropriate extraction process for Huoxue San was investigated. Single-factor experiments were conducted at different levels with the material-liquid ratio, ethanol concentration, extraction time, number of extractions, and soaking time as influencing factors. The total content of six components (demethylene berberine, palmatine hydrochloride, vanillic acid, luteolin, apigenin, and baicalin) was used as the evaluation index to screen out the main factors affecting the extraction of Huoxue San and determine the optimal level. The experimental results are shown in Tables 1-5.

[0039] Table 1. Effect of solid-liquid ratio (times the amount of ethanol) on the extraction of Huoxue San (a traditional Chinese medicine formula).

[0040]

[0041] Table 2. Effect of ethanol concentration on the extraction of Huoxue San (a traditional Chinese medicine formula).

[0042]

[0043] Table 3. Effect of extraction time on the extraction of Huoxue San (a traditional Chinese medicine formula).

[0044]

[0045] Table 4. Effect of extraction times on the extraction of Huoxue San (a traditional Chinese medicine formula).

[0046]

[0047] Table 5 Effect of soaking time on the extraction of Huoxue San (a traditional Chinese medicine formula).

[0048]

[0049] Experimental results show that the optimal extraction process for Huoxue San is as follows: ethanol concentration: 75%; extraction time: 30 min; number of extractions: 2; material-to-liquid ratio: 14 times; soaking time: 50 min.

[0050] The chromatographic conditions for determining the content of the six components are as follows:

[0051] Chromatographic column: YMC-Pack ODS-A C18-001 (250 mm × 4.6 mm, 5 μm); mobile phase A: (0.1% formic acid-water solution), mobile phase B: (acetonitrile), gradient elution; flow rate: 1 mL / min; column temperature: 30 ℃; detection wavelength: 280 nm; injection volume: 10 μL. The gradient elution program is shown in Table 6 below:

[0052] Table 6 Gradient elution program

[0053]

[0054] The linear equations for the six main components are as follows: vanillic acid: y = 11751x + 5211.8; demethylene berberine: y = 21257x + 4583.8; palmatine hydrochloride: y = 40713x - 103853; luteolin: y = 21509x + 24238; apigenin: y = 33616x + 5788; baicalin: y = 67321x + 33210.

[0055] Example 3: Preparation of freeze-dried powder for promoting blood circulation:

[0056] Take the extract of Huoxue San, filter it, make up to 250 mL, shake well, transfer it to a round bottom flask and rotary evaporate until it becomes viscous (not sticking to the wall). Transfer it to an evaporating dish (with a layer of plastic wrap at the bottom), heat it in a water bath (80 ℃) until it becomes viscous, cover it with plastic wrap, freeze it overnight (-18 ℃), and then freeze it in a freeze dryer (turn on the switch, press the refrigeration button, wait for the temperature to reach -35 ℃, put in the sample, press the vacuum pump and vacuum gauge, wait until the vacuum state reaches a few degrees, and then turn off the vacuum pump). Freeze-dry for three days and three nights, take the freeze-dried powder of Huoxue San, grind it into a fine powder in a mortar and pestle, and you have obtained the product.

[0057] Example 4: Study on the preparation process of Huoxue San gel plaster - ratio of phase A to phase B

[0058] Sensory evaluation and initial tack of the gel patch were used as evaluation indicators to comprehensively score the preparation process parameters of the Huoxue San gel patch. Specific scoring criteria are shown in Table 7.

[0059] Table 7. Gel Patch Scoring Criteria

[0060]

[0061] (1) Initial viscosity evaluation method

[0062] The initial tack of the adhesive plaster was determined according to the method for determining the initial tack of plasters under General Chapter 0952 of Part IV of the 2020 edition of the Chinese Pharmacopoeia. The rolling ball ramp stop method was used to determine the adhesive force of the gel plaster. The method was based on Method I (Initial Tack Determination) of General Chapter 0925 of Part IV of the Chinese Pharmacopoeia (2020 edition). Initial tack was determined using the rolling ball ramp stop method. For each sample, the protective layer was removed, and the sample was placed in the center of an inclined plate 30 cm long with a 30° angle, with the plaster side facing upwards. The upper 10 cm and lower 15 cm of the ramp were covered with a 0.025 mm thick polyester film, leaving a 5 cm plaster surface exposed in the middle. Steel balls of different diameters were rolled freely down the ramp, and the ball number of the largest steel ball that the adhesive surface could adhere to was recorded. A larger ball number indicated better initial tack.

[0063] (2) Peel strength evaluation method

[0064] According to the test standard YY / T0148-2006 for the peel strength of medical tape, the gel patch was applied to a clean phenolic resin board and pressed with a 2 kg roller at 600 mm / min. -1 Roll at a speed of 4 round trips, and after placing for 10 minutes, pull the force gauge hook at 300 mm / min. -1 Pull the material away from the phenolic resin board at a speed of 180° and then turn it back at a 180° angle. Record the reading displayed on the force gauge. Select the average value of a range of data with a standard deviation within 5% as the peel strength value. Measure this three times consecutively and take the average value.

[0065] (3) Comprehensive sensory indicators

[0066] The comprehensive sensory evaluation index assesses aspects such as the uniformity, spreadability, skin adherence, film residue, permeation, and reapplied performance of the gel patch.

[0067] Investigate the effect of the ratio of phase A to phase B on the formation of Huoxue San plaster:

[0068] Table 8. Analysis of the ratio of phase A to phase B

[0069]

[0070] Sample 1: Weigh 0.5g of aluminum hydroxyl, 3g of NP-700, and 35g of glycerin, mix them, and stir at 200 rpm for 3 minutes at room temperature using a high-shear mixing emulsifier (model: BME100LX) to obtain phase A. Weigh 2g of PVPK90, 0.5g of tartaric acid, and 20g of Wahaha purified water, mix them, and let them stand overnight at room temperature to obtain phase B. Mix phase B with phase C (blood-activating powder extract) to obtain phase D. Mix phase A with phase D using a high-shear mixing emulsifier (model: BME100LX) to obtain the final coating. Maintain the mixing temperature at approximately 30°C to keep it soft. Take the above coating, place it in the center of a non-woven fabric, cover it with a pre-cut transparent plastic cover, and flatten it with a glass rod to obtain the final product.

[0071] Sample 2: Weigh 0.25 g of aluminum hydroxyl, 3 g of NP-700, and 20 g of glycerin (mixed with a high-shear emulsifier, model: BME100LX, stirred at 200 rpm for 3 min at room temperature) as phase A. Weigh 2 g of PVPK90, 0.5 g of tartaric acid, and 20 g of Wahaha purified water, mix, and let stand at room temperature overnight as phase B. Mix phase B with phase C (blood-activating powder extract) as phase D. Mix phase A with phase D (high-shear emulsifier, model: BME100LX) to obtain the final coating. Maintain the mixing temperature at approximately 30 ℃ to keep it soft. Take the above coating, place it in the center of a non-woven fabric, place a pre-cut transparent plastic cover underneath, maintain the mixing temperature at approximately 30 ℃ to keep it soft, cover with the pre-cut transparent plastic cover, and flatten with a glass rod to obtain the final product.

[0072] Sample 3: Weigh 0.5 g of aluminum hydroxyl, 3 g of NP-700, and 35 g of glycerin (mixed with a high-shear emulsifier, model: BME100LX, stirred at 200 rpm for 3 min at room temperature) as phase A. Weigh 4 g of PVPK90, 0.5 g of tartaric acid, and 40 g of Wahaha purified water, mix, and let stand at room temperature overnight as phase B. Mix phase B with phase C (blood-activating powder extract) as phase D. Mix phase A and phase D with a high-shear emulsifier (model: BME100LX) to obtain the final coating. Maintain the mixing temperature at approximately 30 ℃ to keep it soft. Take the above coating, place it in the center of a non-woven fabric, place a pre-cut transparent plastic cover underneath, maintain the mixing temperature at approximately 30 ℃ to keep it soft, cover with the pre-cut transparent plastic cover, and flatten with a glass rod to obtain the final product.

[0073] Sample rating:

[0074] Uniformity score (Sample 1: 8 points; Sample 2: 8 points; Sample 3: 8 points); Spreadability (Sample 1: 6 points; Sample 2: 6 points; Sample 3: 6 points); Spreadability (Sample 1: 7 points; Sample 2: 8 points; Sample 3: 7 points); Skin followability (Sample 1: 7 points; Sample 2: 8 points; Sample 3: 7 points); Film residue (Sample 1: 7 points; Sample 2: 7 points; Sample 3: 7 points); Repeated peeling / removal performance (Sample 1: 7 points; Sample 2: 8 points; Sample 3: 8 points); (7 points); Initial tack of Sample 1: 15# steel ball; Initial tack of Sample 2: 15# steel ball; Initial tack of Sample 3: 13# steel ball. Therefore, the initial tack score for Sample 1 is 15 / 15*20 = 20; the initial tack score for Sample 2 is 15 / 15*20 = 20; the initial tack score for Sample 3 is 13 / 15*20 = 17; the average peel strength of Sample 1 over three tests is 1.722 N; the average peel strength of Sample 2 over three tests is 1.8623 N; the average peel strength of Sample 3 over three tests is 1.7530 N. N, i.e., the peel strength score of sample 1 = 1.722 / 1.8623*20 = 18; the peel strength score of sample 2 = 1.8623 / 1.8623*20 = 20; the peel strength score of sample 3 = 1.7530 / 1.8623*20 = 19; the total score of sample 1 is 80 points, the total score of sample 2 is 85 points, and the total score of sample 3 is 78 points. According to the total score, it can be seen that under condition 2, that is, the ratio of A:B is 1:1, the blood-activating powder gel plaster has the best molding effect. The results are shown in Table 9.

[0075] Table 9. Overall Score of Auxiliary Material Usage Ratio

[0076]

[0077] Example 5: Study on the preparation process of Huoxue San gel plaster - selection of C-phase Huoxue San extract and lyophilized powder

[0078] The effects of freeze-dried powder and extract of Huoxuesan on the formation of Huoxuesan plaster were investigated.

[0079] Table 10. Evaluation of the molding effects of freeze-dried powder and extract of Huoxuesan.

[0080]

[0081] Sample 1: Weigh 0.25 g of aluminum hydroxyl, 1.5 g of NP-700, and 5 g of glycerin, mix them (using a high-shear mixing emulsifier, model: BME100LX), and stir at 200 rpm for 3 min at room temperature to obtain phase A. Weigh 2 g of PVPK90, 0.25 g of tartaric acid, and 5 g of Wahaha purified water, mix them, and let them stand overnight at room temperature to obtain phase B. Mix phase B with phase C (freeze-dried Huoxue San powder) to obtain phase D. Mix phase A with phase D (using a high-shear mixing emulsifier, model: BME100LX), and stir at 200 rpm for 3 min at room temperature to obtain the final coating. Maintain the temperature at approximately 30 ℃ to keep it soft. Take the above coating, place it in the center of a nonwoven fabric, cover it with a pre-cut transparent plastic cover, and flatten it with a glass rod to obtain the final product.

[0082] Sample 2: Weigh 0.5 g of aluminum hydroxyl, 3 g of NP-700, and 10 g of glycerin, mix them (using a high-shear mixing emulsifier, model: BME100LX), and stir at 200 rpm for 3 min at room temperature to obtain phase A. Weigh 4 g of PVPK90, 0.5 g of tartaric acid, and 10 g of Wahaha purified water, mix them, and let them stand overnight at room temperature to obtain phase B. Mix phase B with phase C (blood-activating powder extract) to obtain phase D. Mix phase A with phase D (using a high-shear mixing emulsifier, model: BME100LX), and stir at 200 rpm for 3 min at room temperature to obtain the final coating. Maintain the temperature at approximately 30 ℃ to keep it soft. Take the above coating, place it in the center of a non-woven fabric, cover it with a pre-cut transparent plastic cover, and flatten it with a glass rod to obtain the final product.

[0083] Uniformity score (Sample 1: 9 points; Sample 2: 2 points); Spreadability (Sample 1: 9 points; Sample 2: 1 point); Spreadability (Sample 1: 8 points; Sample 2: 2 points); Skin adherence (Sample 1: 8 points; Sample 2: 3 points); Film residue (Sample 1: 7 points; Sample 2: 2 points); Repeated application / removal performance (Sample 1: 7 points; Sample 2: 2 points). (3 points); Initial tack of Sample 1: No. 3 steel ball; Initial tack of Sample 2: No. 5 steel ball; That is, the initial tack score of Sample 1 = 3 / 5 * 20 = 12; the initial tack score of Sample 2 = 5 / 5 * 20 = 20; The average of the three peel strengths of Sample 1 is 2.5313 N; The average of the three peel strengths of Sample 2 is 11.7837 N; That is, the peel strength score of Sample 1 = 2.5313 / 11.7837 * 20 = 4; The peel strength score of Sample 2 = 11.7837 / 11.7837 * 20 = 20; The total score of Sample 1 is 64 points, and the total score of Sample 2 is 53 points. Therefore, according to the total score, under condition 1, that is, under the condition of using Huoxuesan freeze-dried powder, the Huoxuesan gel plaster has the best molding effect. The results are shown in Table 11.

[0084] Table 11 Comprehensive score for evaluating the molding effect of Huoxuesan freeze-dried powder and Huoxuesan extract

[0085]

[0086] Example 6: Study on the preparation process of Huoxue San gel plaster – Dosage of PVPK90

[0087] The effect of PVPK90 dosage on the formation of Huoxue San plaster was investigated.

[0088] Table 12 PVPK90 Dosage Study

[0089]

[0090] Sample 1: Weigh 0.25 g of aluminum hydroxyl, 1.5 g of NP-700, and 5 g of glycerin, mix them (using a high-shear mixing emulsifier, model: BME100LX), and stir at 200 rpm for 3 min at room temperature to obtain phase A. Weigh 2 g of PVPK90, 0.25 g of tartaric acid, and 5 g of Wahaha purified water, mix them, and let them stand overnight at room temperature to obtain phase B. Mix phase B with phase C (blood-activating powder extract) to obtain phase D. Mix phase A with phase D (using a high-shear mixing emulsifier, model: BME100LX), and stir at 200 rpm for 3 min at room temperature to obtain the final coating. Maintain the temperature at approximately 30 ℃ to keep it soft. Take the above coating, place it in the center of a non-woven fabric, cover it with a pre-cut transparent plastic cover, and flatten it with a glass rod to obtain the final product.

[0091] Sample 2: Weigh 0.25 g of aluminum hydroxyl, 1.5 g of NP-700, and 5 g of glycerin, mix them (using a high-shear mixing emulsifier, model: BME100LX), and stir at 200 rpm for 3 min at room temperature to obtain phase A. Weigh 1.8 g of PVPK90, 0.25 g of tartaric acid, and 5 g of Wahaha purified water, mix them, and let them stand overnight at room temperature to obtain phase B. Mix phase B with phase C (blood-activating powder extract) to obtain phase D. Mix phase A with phase D (using a high-shear mixing emulsifier, model: BME100LX), and stir at 200 rpm for 3 min at room temperature to obtain the final coating. Maintain the temperature at approximately 30 ℃ to keep it soft. Take the above coating, place it in the center of a non-woven fabric, cover it with a pre-cut transparent plastic cover, and flatten it with a glass rod to obtain the final product.

[0092] Sample 3: Weigh 0.25 g of aluminum hydroxyl, 1.5 g of NP-700, and 5 g of glycerin, mix them (using a high-shear mixing emulsifier, model: BME100LX), and stir at 200 rpm for 3 min at room temperature to obtain phase A. Weigh 2.2 g of PVPK90, 0.25 g of tartaric acid, and 5 g of Wahaha purified water, mix them, and let them stand at room temperature overnight to obtain phase B. Mix phase B with phase C (blood-activating powder extract) to obtain phase D. Mix phase A with phase D (using a high-shear mixing emulsifier, model: BME100LX), and stir at 200 rpm for 3 min at room temperature to obtain the final coating. Maintain the temperature at approximately 30 ℃ to keep it soft. Take the above coating, place it in the center of a non-woven fabric, cover it with a pre-cut transparent plastic cover, and flatten it with a glass rod to obtain the final product.

[0093] Uniformity score (Sample 1: 8 points; Sample 2: 8 points; Sample 3: 8 points); Spreadability (Sample 1: 6 points; Sample 2: 6 points; Sample 3: 6 points); Spreadability (Sample 1: 7 points; Sample 2: 8 points; Sample 3: 7 points); Skin adhesion (Sample 1: 7 points; Sample 2: 8 points; Sample 3: 7 points); Film residue (Sample 1: 7 points; Sample 2: 7 points; Sample 3: 7 points); Repeated peeling and sticking performance (Sample 1: 7 points; Sample 2: 8 points; Sample 3: 7 points); Initial tack of Sample 1: 5 steel ball; Initial tack of Sample 2: 7 steel ball; Initial tack of Sample 3: 7 steel ball; i.e., Initial tack score of Sample 1 = 5 / 7 * 20 = 14; Initial tack score of Sample 2 = 7 / 7 * 20 = 20; Initial tack score of Sample 3 = 7 / 7 * 20 = 20. The average peel strength of Sample 1 was 5.1643 N; the average peel strength of Sample 2 was 5.1893 N; and the average peel strength of Sample 3 was 5.5327 N. Therefore, the peel strength score for Sample 1 was 5.1643 / 5.5327*20 = 18; the peel strength score for Sample 2 was 5.1893 / 5.5327*20 = 18; and the peel strength score for Sample 3 was 5.5327 / 5.5327*20 = 20. The total score for Sample 1 was 74 points, for Sample 2 it was 83 points, and for Sample 3 it was 82 points. Based on the total scores, it can be concluded that condition 3, i.e., PVPK90 at 2.2g, resulted in the best molding effect for the Huoxue San gel plaster. The results are shown in Table 13.

[0094] Table 13 Overall Rating of PVPK90 Dosage

[0095]

[0096] Example 7 Study on the preparation process of Huoxue San gel plaster - Tartaric acid dosage

[0097] The effect of tartaric acid dosage on the formation of Huoxue San plaster was investigated.

[0098] Table 14 Investigation on Tartaric Acid Dosage

[0099]

[0100] Sample 1: Weigh 0.25 g of aluminum hydroxyl, 1.5 g of NP-700, and 5 g of glycerin, mix them, and stir at 200 rpm for 3 min at room temperature using a high-shear mixing emulsifier (model: BME100LX) to obtain phase A. Weigh 2 g of PVPK90, 0.25 g of tartaric acid, and 5 g of Wahaha purified water, mix them, and let them stand overnight at room temperature to obtain phase B. Mix phase B with phase C (freeze-dried Huoxuesan powder) to obtain phase D. Mix phase A with phase D (using a high-shear mixing emulsifier (model: BME100LX)) and stir at 200 rpm for 3 min at room temperature to obtain the final coating. Maintain the temperature at approximately 30 ℃ to keep it soft. Take the above coating, place it in the center of a nonwoven fabric, cover it with a pre-cut transparent plastic cover, and flatten it with a glass rod to obtain the final product.

[0101] Sample 2: Take 0.25 g of aluminum hydroxyl, 1.5 g of NP-700, and 5 g of glycerin, mix them (using a high-shear mixing emulsifier, model: BME100LX), and stir at 200 rpm for 3 min at room temperature to obtain phase A. Weigh 2 g of PVPK90, 0.24 g of tartaric acid, and 5 g of Wahaha purified water, mix them, and let them stand overnight at room temperature to obtain phase B. Mix phase B with phase C (blood-activating powder extract) to obtain phase D. Mix phase A with phase D (using a high-shear mixing emulsifier, model: BME100LX), and stir at 200 rpm for 3 min at room temperature to obtain the final coating. Maintain the temperature at approximately 30 ℃ to keep it soft. Take the above coating, place it in the center of a non-woven fabric, cover it with a pre-cut transparent plastic cover, and flatten it with a glass rod to obtain the final product.

[0102] Uniformity score (Sample 1: 8 points; Sample 2: 8 points); Spreadability (Sample 1: 8 points; Sample 2: 8 points); Skin adherence (Sample 1: 8 points; Sample 2: 7 points); Film residue (Sample 1: 9 points; Sample 2: 8 points); Repeated application / removal performance (Sample 1: 9 points; Sample 2: 7 points); (8 points); Initial adhesion of Sample 1: No. 6 steel ball; Initial adhesion of Sample 2: No. 6 steel ball; That is, the initial adhesion score of Sample 1 = 6 / 6*20 = 20; The initial adhesion score of Sample 2 = 6 / 6*20 = 20; The average of the three peel strengths of Sample 1 is 5.4583N; The average of the three peel strengths of Sample 2 is 5.1853N; That is, the peel strength score of Sample 1 = 5.4583 / 5.4583*20 = 20; The peel strength score of Sample 2 = 5.1853 / 5.4583*20 = 19; The total score of Sample 1 is 90 points, and the total score of Sample 2 is 85 points. Therefore, according to the total score, it can be seen that under condition 1, that is, under the condition of tartaric acid of 0.25g, the blood circulation powder gel plaster has the best molding effect. The results are shown in Table 15.

[0103] Table 15 Overall Score for Tartaric Acid Dosage

[0104]

[0105] Example 8: Study on the preparation process of Huoxue San gel plaster – dosage of aluminum hydroxyacetate and NP-700

[0106] The effects of the dosage of aluminum hydroxide and NP-700 on the formation of Huoxue San plaster were investigated.

[0107] Table 16. Dosage Investigation of Aluminum Hydroxyhydroxymethylpyrazine and NP-700

[0108]

[0109] Sample 1: Weigh 0.25 g of aluminum hydroxyl, 1.5 g of NP-700, and 5 g of glycerin. Mix them using a high-shear mixing emulsifier (model: BME100LX) and stir at 200 rpm for 3 min at room temperature to obtain phase A. Weigh 2.2 g of PVPK90, 0.25 g of tartaric acid, and 5 g of Wahaha purified water. Mix them and let stand overnight at room temperature to obtain phase B. Mix phase B with phase C (freeze-dried Huoxue San powder) to obtain phase D. Mix phase A with phase D (high-shear mixing emulsifier, model: BME100LX) and stir at 200 rpm for 3 min at room temperature to obtain the final coating. Maintain the temperature at approximately 30 ℃ to keep it soft. Place the coating in the center of a nonwoven fabric, cover it with a pre-cut transparent plastic cover, and flatten it with a glass rod to obtain the final product.

[0110] Sample 2: Weigh 0.75 g of aluminum hydroxyl, 4.5 g of NP-700, and 20 g of glycerin, mix them, and stir at 200 rpm for 3 min at room temperature using a high-shear mixing emulsifier (model: BME100LX) to obtain phase A. Weigh 8.8 g of PVPK90, 1 g of tartaric acid, and 20 g of Wahaha purified water, mix them, and let them stand overnight at room temperature to obtain phase B. Mix phase B with phase C (blood-activating powder extract) to obtain phase D. Mix phase A with phase D (high-shear mixing emulsifier, model: BME100LX), and stir at 200 rpm for 3 min at room temperature to obtain the final coating. Maintain the temperature at approximately 30 ℃ to keep it soft. Take the above coating, place it in the center of a non-woven fabric, cover it with a pre-cut transparent plastic cover, and flatten it with a glass rod to obtain the final product.

[0111] Uniformity score (Sample 1: 8 points; Sample 2: 8 points); Spreadability (Sample 1: 8 points; Sample 2: 8 points); Skin adherence (Sample 1: 8 points; Sample 2: 7 points); Film residue (Sample 1: 9 points; Sample 2: 8 points); Repeated peeling / removal performance (Sample 1: 8 points; Sample 2: 8 points); (9 points); Initial tack of Sample 1: No. 4 steel ball; Initial tack of Sample 2: No. 8 steel ball; That is, the initial tack score of Sample 1 = 4 / 8 * 20 = 10; the initial tack score of Sample 2 = 8 / 8 * 20 = 20; The average of the three peel strengths of Sample 1 is 5.5327 N; The average of the three peel strengths of Sample 2 is 5.6113 N; That is, the peel strength score of Sample 1 = 5.5327 / 5.6113 * 20 = 19; The peel strength score of Sample 2 = 5.6113 / 5.6113 * 20 = 20; The total score of Sample 1 is 76 points, and the total score of Sample 2 is 91 points. Therefore, according to the total score, under condition 2, that is, under the condition of 0.75 g of aluminum hydroxide and 4.5 g of NP-700, the blood-activating powder gel plaster has the best molding effect. The results are shown in Table 17.

[0112] Table 17 Comprehensive Evaluation Table for Dosage of Aluminum Hydroxyhydroxymethylamine and NP-700

[0113]

[0114] In summary, based on Examples 4-8, the final preparation process for the Huoxue San gel patch is determined as follows:

[0115] Phase A: Weigh out 0.75 g of aluminum hydroxyl, 4.5 g of NP-700, and 20 g of glycerin, and mix them (using a high-shear emulsifier, model: BME100LX) at room temperature and stir at 200 r for 3 min.

[0116] Phase B: Weigh out 8.8 g of PVPK90, 1 g of tartaric acid, and 20 g of Wahaha purified water, mix them, and let stand at room temperature overnight.

[0117] Phase C: Take 8 g of lyophilized Huoxue San powder, add 4 mL of 75% ethanol, sonicate for 8 min, then add another 4 mL of 75% ethanol and sonicate for 8 min to obtain the final product.

[0118] Phase D: Weigh 18.2 g of Phase B and mix it with Phase C (using a high-shear emulsifier, model: BME100LX), and stir at 200 r for 3 min at room temperature.

[0119] Take 7.85 g of phase A and 12.6 g of phase D, mix them (high shear emulsifier, model: BME100LX), stir at 200 r for 3 min at room temperature to obtain the final coating, and keep the temperature at about 30 ℃ to maintain its soft state.

[0120] Take 0.25 g of the above coating material, place it in the center of a non-woven fabric, cover it with a pre-cut transparent plastic cover, and flatten it with a glass rod to obtain the blood circulation gel plaster. Store it in a sealed bag at room temperature.

[0121] The prescription ratios are as follows:

[0122] Table 18 Prescription Proportions

[0123]

[0124] Example 9: Pharmacodynamic Study of Blood-Activating Powder Gel Plaster – Comparison of Soft Tissue Injuries

[0125] This study investigated the effects of traditional Huoxue San powder and Huoxue San plaster on soft tissue injury.

[0126] Hair was shaved from the left and right legs of SD rats. Soft tissue injury was induced by striking the rats with a heavy object. A 200g weight was dropped freely from a fixed height, striking the lateral muscular areas of the rats 10 times to establish an acute soft tissue injury model. The rats were visually observed for scattered hemorrhages, bruising, and swelling under the skin of their thighs. A successful model was considered established if there were no obvious skin ruptures, joint dislocations, fractures, or other open injuries, and if all other areas were normal.

[0127] Thirty rats were randomly divided into four groups: a blank control group, a model group, a positive control group (Xueshan Jinluohan group), a plaster group, and a powder group, with six rats in each group. Significant differences in appearance scores and body weight were ensured among the groups. The medication was applied to the injured area of ​​the right limb and secured with medical tape, with continuous monitoring for any dislodgement. The Xueshan Jinluohan group received 0.1 g / kg of medication; the Huoxue San powder and the novel plaster group received 0.9 g / kg of medication. All groups received the medication once daily for approximately 4 hours each time, for 7 consecutive days.

[0128] The degree of swelling at the injury site was observed in each group of rats before modeling and on days 1, 3, 5, and 7 after drug administration. The circumference of the injury site was measured using a soft measuring tape, and the degree of swelling was calculated. Swelling degree (%) = (measured circumference / circumference before modeling - 1) × 100%. The appearance of the damaged soft tissue of rats was scored on days 1, 3, 5, and 7 during the drug administration period, and the changes in appearance were recorded by photograph. The appearance scoring criteria for rat soft tissue injury are shown in Table 19.

[0129] Table 19 Appearance Scoring Table for Soft Tissue Injury in Rats

[0130]

[0131] On day 1 of administration, compared with the control group, the swelling degree of rats in the model group was significantly increased (P<0.05). On day 3 of administration, compared with the control group, the swelling degree of rats in the model group was significantly increased (P<0.05); compared with the model group, the swelling degree of rats in each dose of Huoxue Sansan (a traditional Chinese medicine) and the plaster group was significantly decreased (P<0.05). On day 5 of administration, compared with the control group, the swelling degree of rats in the model group was significantly increased (P<0.05); compared with the model group, the swelling degree of rats in each dose of Huoxue Sansan and the plaster group was significantly decreased (P<0.05). On day 7 of administration, compared with the control group, the swelling degree of rats in the model group was significantly increased (P<0.05), while the swelling degree of rats in each dose of Huoxue Sansan and the plaster group was significantly decreased (P<0.05). The results are shown in Table 20.

[0132] Table 20 Comparison of swelling degree in rats of each group on days 1, 3, 5, and 7 after drug administration ( ±s, n=6)

[0133]

[0134] On day 1 of administration, the appearance scores of the model group and all administered groups were significantly higher than those of the control group (P < 0.01); on day 3 of administration, the appearance scores of all administered groups were significantly higher than those of the model group (P < 0.01); on day 5 of administration, the appearance scores of all administered groups were significantly higher than those of the model group (P < 0.05); on day 7 of administration, except for the low-dose powder group, the appearance scores of all administered groups were significantly higher than those of the model group (P < 0.05). The results are shown in Table 21.

[0135] Table 21 Soft tissue injury index scores of rats in each group ( ±s, n=6)

[0136]

[0137] Images of the right hind limb of rats in each group at days 1, 3, 5, and 7 were collected to observe apparent soft tissue injury and compare the pharmacodynamics of the traditional Huoxue San powder and the Huoxue San gel plaster. In the control group, only mild ecchymosis was observed in the early stages of modeling, which recovered rapidly over time. In the model group, the injury was severe, with slow resolution of swelling and ecchymosis; significant pigmentation remained on day 7, indicating delayed recovery even with successful modeling and no intervention. The positive control group showed significantly better recovery than the model group, with skin essentially returning to normal by day 7. Both the Huoxue San powder and plaster groups significantly improved symptoms of ecchymosis and swelling in the affected limb; from day 3, the color of the ecchymosis became noticeably lighter, and the swelling gradually subsided. By day 7, skin color and hair growth were close to those of the control group, and the overall recovery process was not significantly different from the positive control group. This suggests that both formulations of Huoxue San have good therapeutic effects on acute soft tissue injury, with comparable efficacy and both effectively promoting injury repair. Results are shown below. Figure 5 .

[0138] On day 7 of drug administration, subcutaneous muscle and soft tissue samples were collected from rats in each group along the injury site, centered on the site of impact. The samples were rinsed in 0.9% sodium chloride solution, dried with filter paper, weighed using an electronic balance, and stored at -80 ℃. For measurement, the tissue was removed, homogenized on ice with 0.9% sodium chloride solution at 4 ℃, and centrifuged at 3000 r / min for 10 min (centrifugation radius 10 cm). The supernatant was collected. TNF-α, IL-1β, PGE2, and IL-6 levels in the local tissue were measured using ELISA. Results are shown below. Figure 1 .

[0139] Rat tissue samples were removed from the fixative, fixed, dehydrated, paraffin-embedded, and cut into 4 µm thick sections. The sections were spread and heated to ensure firm adhesion between the tissue and the slide. The tissue sections underwent routine dewaxing, staining with hematoxylin and eosin, dehydrating with graded ethanol, clearing with xylene, and mounting with neutral resin. The pathological changes in the soft tissues of each group of rats were observed under a microscope. Images of tissue lesions at fixed locations were taken, and the pathological changes in the rat soft tissues were analyzed.

[0140] In the normal control group, rat muscle cells were well-organized with clear striations, and showed no degeneration or necrosis. The interstitium showed no congestion or edema, no inflammatory cell infiltration, and no fibroblast proliferation or other lesions. In the model group, rat muscle cells showed mild to moderate inflammatory cell infiltration and edema, with some muscle cells ruptured and necrotic. All drug-treated groups showed varying degrees of improvement in muscle tissue lesions compared to the model group, with significantly reduced muscle lesions, smaller lesion area, milder muscle cell degeneration, and slight inflammatory cell infiltration in the interstitium. Results are shown below. Figure 2 .

[0141] Example 10 Pharmacodynamic Study of Blood Circulation Powder Gel Plaster – Comparative Study of its Efficacy in Osteoarthritis

[0142] Twenty-four SD rats were randomly divided into four groups (n=6 per group): a control group, a model group, a blood-activating powder group, and a blood-activating powder plaster group. Except for the control group, all other groups were anesthetized with intraperitoneal injection of 1% sodium pentobarbital. The hair around the right knee joint was shaved, and the skin was disinfected with 75% ethanol. The right knee was slightly bent, and 0.2 mL of 4% papain solution was injected into the right knee joint cavity approximately 0.5 cm above and outside the patellar ligament attachment point. This injection was repeated every 3 days for 3 consecutive days. Behavioral changes were observed after 7 days. Successful modeling was indicated by contraction or lameness in the right hind limb. The control group rats received an equal volume of saline injected into the right knee joint cavity. Based on clinical dosage conversion, the blood-activating powder group rats were treated with a 0.9 g / kg dose of a mixture of maltose and applied to the right hind limb, wrapped with gauze, once daily for 4 hours each time, for 28 days.

[0143] Rats were placed in an activity box for 5–6 minutes to acclimatize. While the rats were at rest, a 0.8 mm diameter stimulation needle was inserted through the stainless steel mesh at the bottom of the activity box to gradually apply pressure to the rats' paws until they exhibited a clear paw retraction / leg lifting response. The maximum pressure was recorded. Each rat was stimulated 5 times, with a 30-second interval between each stimulation. The results are shown in Table 22.

[0144] Table 22 Rat mechanical reflex thresholds ( ±s, n=6)

[0145]

[0146] Two parallel walkways, each 110 cm long, 20 cm wide, and 20 cm high, were constructed using cardboard. The walkways were open at the top, with entrances and exits at both ends. Cameras were placed at appropriate locations to record the experimental process in real time. Xuan paper and calligraphy ink were used for footprint recording. Rats underwent 4-5 days of acclimatization training before the formal experiment. In the formal experiment, a rat walked the entire walkway normally from the starting point, leaving 5-6 pairs of consecutive footprints, constituting one valid experiment. Each rat needed to complete 1-3 valid experiments. The footprints on the Xuan paper were converted into electronic images using a scanner, and the data was read using ImageJ software. The results are shown in Table 23.

[0147] Table 23 Rat Gait Behavior Scoring Table ( ±s, n=6)

[0148]

[0149] To observe the pathological morphology of knee joint cartilage tissue, rats were anesthetized and sacrificed. The fur at the knee joint was removed, and the knee joint cartilage was completely harvested. The cartilage tissue was collected, fixed overnight in 4% paraformaldehyde, decalcified in EDTA, embedded in paraffin, cut into 5μm tissue sections, stained with hematoxylin and eosin (HE), and observed and images were acquired under an optical microscope.

[0150] The synovium in the blank control group consisted of several layers of flattened synovial cells, with uniform thickness, clear boundaries, and no hyperplasia; the subsynovial connective tissue was loose, with regularly arranged fibers, no edema or congestion, and a few inflammatory cells. In the model group, the synovium was significantly thickened and hyperplastic, with an increased number of synovial cell layers; the subsynovial connective tissue showed edema and congestion (increased red areas), disordered and broken fiber structure, and extensive infiltration of inflammatory cells. In the powder and plaster groups, the synovial cells were arranged relatively regularly, with only a small number of cells stacked in localized areas; inflammatory cells were significantly reduced, the synovial thickness was close to that of the blank group, subsynovial connective tissue edema had basically disappeared, and the fiber arrangement tended to be regular. Results are shown below. Figure 3 .

[0151] One hour after the last administration, blood was collected from the abdominal aorta of anesthetized rats. The blood was centrifuged at 3000 r / min for 10 min, and the supernatant was collected and stored at -80℃. A suitable amount of rat serum was collected, and the levels of TNF-α, IFN-γ, L-1β, and IL-17 were detected according to the ELISA kit instructions. Compared with the blank control group, the levels of TNF-α, IFN-γ, IL-1β, and IL-17 in the knee joint of rats in the model group were significantly increased (**P < 0.01); compared with the model group, the levels of TNF-α, IFN-γ, IL-1β, and IL-17 in the knee joint of rats in the powder group and plaster group were significantly decreased (##P < 0.01). Results are shown below. Figure 4 .

Claims

1. A blood-activating and stasis-reducing gel patch, characterized in that, It is composed of freeze-dried Huoxue San powder and a gel matrix; the freeze-dried Huoxue San powder includes seven herbs: Liu Ji Nu, Di Fu Zi, Huang Bai, Ban Zhi Lian, Di Bie Chong, Hu Zhang, and Sheng Nan Xing; the gel matrix contains cross-linked polymers, thickeners, cross-linking agents, cross-linking regulators, and humectants.

2. The blood-activating and stasis-removing gel patch according to claim 1, characterized in that, The crosslinked polymer is a partially neutralized polyacrylic acid compound, wherein the partially neutralized compound is at least one of NP-600 with a neutralization degree of 70%, NP-700 with a neutralization degree of 50%, and NP-800 with a neutralization degree of 35%; the tackifier is at least one of sodium carboxymethyl cellulose, sodium methyl cellulose, polyvinyl alcohol, polyvinylpyrrolidone K90, gelatin, and gum arabic; the crosslinking agent is at least one of aluminum hydroxyl, aluminum hydroxide, aluminum chloride, aluminum sulfate, and alum; the crosslinking regulator is at least one of tartaric acid, citric acid, lactic acid, EDTA, or EDTA-2Na; and the humectant is at least one of propylene glycol, glycerin, 1,3-butanediol, and polyethylene glycol.

3. The blood-activating and stasis-removing gel patch according to claim 1 or 2, characterized in that, The weight parts of each component in the gel matrix are as follows: 4.5 parts of cross-linked polymer, 8.8 parts of thickener, 0.75 parts of cross-linking agent, 1 part of cross-linking regulator, 20 parts of humectant, and 20 parts of purified water; the weight parts of the blood-activating powder lyophilized powder are 2 parts.

4. The blood-activating and stasis-removing gel patch according to claim 1, characterized in that, The weight ratio of the following herbs is 170:170:160:170:170:85:

75.

5. The blood-activating and stasis-removing gel plaster according to claim 1, characterized in that, The blood-activating powder freeze-dried powder is prepared by the following method: take seven herbs, namely, Artemisia capillaris, Kochia scoparia, Phellodendron chinense, Scutellaria barbata, Eupolyphaga sinensis, Polygonum cuspidatum, and Arisaema heterophyllum, and extract them twice by reflux with 75% ethanol as solvent, 14 times the amount of ethanol each time for 30 minutes. Combine the extracts, concentrate them, and freeze-dry them to obtain the final product.

6. A method for preparing the blood-activating and stasis-removing gel plaster according to any one of claims 1-5, characterized in that, The process includes the following steps: (1) Add crosslinking agent and crosslinking polymer to humectant and mix evenly to obtain phase A; (2) Dissolve thickener and crosslinking regulator in purified water and let stand overnight to obtain phase B; (3) Dissolve blood-activating powder freeze-dried powder in ethanol to obtain phase C; (4) Add phase C to phase B and mix evenly to obtain phase D; then add phase D to phase A, mix evenly, and apply to obtain blood-activating powder gel plaster.

7. The method according to claim 6, characterized in that, The mixing conditions in step (1) are room temperature and stirring at 200 rpm for 3 min; the mixing conditions in step (4) are room temperature and stirring at 200 rpm for 3 min.

8. The method according to claim 6, characterized in that, The ethanol mentioned in step (3) is 75% ethanol, and the mass-volume ratio of Huoxue San freeze-dried powder to ethanol is 1g:1mL.

9. The use of the blood-activating and stasis-removing gel plaster according to any one of claims 1-5 in the preparation of a medicament for treating bruises and sprains.

10. The application according to claim 9, characterized in that, The injuries mentioned are soft tissue injuries or osteoarthritis.

Images