A traditional Chinese medicine composition, preparation, preparation method and application for tonifying qi, blood, yin and yang

Abstract

This invention relates to the field of pharmaceutical preparation technology, specifically to a traditional Chinese medicine composition for tonifying qi, blood, yin, and yang, its preparation method, and its application. The raw materials of the traditional Chinese medicine composition include tortoise shell glue, deer antler glue, donkey-hide gelatin, ginseng, astragalus, codonopsis, and wolfberry. The composition is prepared by fermenting the aforementioned raw materials. The mass ratio of tortoise shell glue, deer antler glue, donkey-hide gelatin, ginseng, astragalus, codonopsis, and wolfberry is 1:0.5-2:0.5-2:1-4:2-5:2-6:2-6. The fermented product obtained by fermenting the raw materials with probiotics shows significant effects in treating qi, blood, yin, and yang deficiency syndromes such as fatigue, myocardial ischemia, cerebral hypoxia, and kidney essence deficiency, and / or in treating and preventing leukopenia caused by bone marrow suppression and / or DNA reduction after radiation exposure and / or low immunity, high-altitude hypoxia, and sudden cardiac death.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical preparation technology, specifically to a traditional Chinese medicine composition, preparation, preparation method, and application for tonifying qi, blood, yin, and yang. Background Technology

[0002] Tonic drugs such as tortoise shell glue, deer antler glue, and donkey-hide gelatin are 80% composed of collagen, amino acids, and peptides, which are cloying and difficult to digest, absorb, and utilize in clinical applications. Although their therapeutic effects are outstanding, they still pose a challenge for weak individuals with insufficient qi and blood. Furthermore, codonopsis, ginseng, and astragalus contain a large number of components that are difficult for the human body to absorb. Existing preparation processes for tonic drugs have the following inherent defects, severely limiting their clinical application and market competitiveness: 1. Low utilization rate of effective components and poor bioavailability: The main components of tortoise shell glue, deer antler glue, and donkey-hide gelatin are large-molecule collagen and its hydrolysates, with relatively large molecular weights. The saponins and polysaccharides in codonopsis, ginseng, and astragalus, as well as the polysaccharides in wolfberry, also have complex structures, making direct absorption in the human body difficult. Traditional water extraction processes cannot fully degrade these large molecules into small-molecule peptides, amino acids, and oligosaccharides that are easily absorbed by the intestines, resulting in difficulty and waste of effective components and overall low bioavailability. 2. Slow onset of action and long duration of action: Due to the low absorption rate of active ingredients, traditional tortoise shell glue, deer antler glue, donkey-hide gelatin, and common tonic formulas often require long-term use to see results, failing to meet modern consumers' demand for highly effective and fast-acting health products. 3. Unpleasant taste and smell, leading to poor patient compliance: The inherent animal odor of tortoise shell glue and deer antler glue, along with the sweet and cloying nature of traditional herbal pastes, makes them difficult for some patients, especially younger people, to accept, reducing medication compliance. 4. Limited functionality and insufficient alignment with modern health needs: Traditional formulas focus on tonification, but their functions in regulating intestinal health and enhancing immunity are insufficiently explored in relation to modern, precise health needs.

[0003] Therefore, developing a pharmaceutical composition specifically for replenishing Qi, blood, Yin, and Yang, and optimizing its formulation and preparation process to obtain a traditional Chinese medicine composition with better absorption, faster onset of action, and more comprehensive functions is of great significance and has become a technical problem urgently needing to be solved in this field. The present invention is proposed based on this background and market demand. Summary of the Invention

[0004] To address the shortcomings of existing technologies, this invention provides a fermented combination of traditional Chinese medicine for replenishing Qi, blood, Yin, and Yang, along with its preparation method and applications. Utilizing probiotic fermentation technology, it solves the technical problems of stagnation and spleen deficiency, and the inability to tolerate tonics. The resulting composition is characterized by rapid absorption and fast onset of action. Through probiotic fermentation, proteins in the medicinal substances are cleaved into small molecule peptides, ginseng and astragalus saponins are transformed into rare ginseng and astragalus saponins, and polysaccharides are cleaved into smaller molecule sugar chains, facilitating the absorption and utilization of the active ingredients. This allows for rapid replenishment of Qi, blood, Yin, and Yang, and is used for conditions of Qi, blood, Yin, and Yang deficiency, such as fatigue, myocardial ischemia, cerebral hypoxia; kidney essence deficiency and blood deficiency leading to lower back and knee weakness, cold limbs, seminal emission, forgetfulness, five-center heat, night sweats; yin-yang deficiency and Qi-blood deficiency after radiotherapy, resulting in leukopenia due to bone marrow suppression; low immunity; high-altitude hypoxia; and prevention of sudden cardiac death.

[0005] This invention is achieved through the following technical solutions:

[0006] The first aspect of the present invention relates to a traditional Chinese medicine composition for replenishing qi, blood, yin, and yang. The raw materials of the traditional Chinese medicine composition include tortoise shell glue, deer antler glue, donkey-hide gelatin, ginseng, astragalus, codonopsis, and wolfberry. The traditional Chinese medicine composition is prepared by fermenting the aforementioned raw materials. The mass ratio of tortoise shell glue, deer antler glue, donkey-hide gelatin, ginseng, astragalus, codonopsis, and wolfberry is 1:0.5-2:0.5-2:1-4:2-5:2-6:2-6.

[0007] Preferably, the fermentation is carried out using probiotics, wherein the probiotics are selected from Lactobacillus plantarum (Lactobacillus plantarum). L. plantarum Lactobacillus acidophilus ( L. acidophilus Lactobacillus casei ( L. casei Lactobacillus rhamnosus ( L.rhamnosus Bacillus licheniformis, Bacillus subtilis Bacillus subtilis ), Lactobacillus acidophilus ( Lactobacillus acidophilus ), Streptococcus thermophilus ( Streptococcus thermophilus Lactobacillus bulgaricus ( Lactobacillus delbrueckii subsp. bulgaricus Any 4 of the following.

[0008] More preferably, the mass ratio of the four probiotics is 1:1:1:1.

[0009] More preferably, the probiotic powder is prepared as a bacterial suspension with a concentration of 1-2 mg / mL; the mass of the bacterial suspension is 1%-2% of the total mass of the raw materials of the traditional Chinese medicine composition.

[0010] Preferably, the raw materials of the traditional Chinese medicine composition further include at least one of the following: Qi-tonifying herbs: red ginseng, American ginseng, and codonopsis; and / or at least one of the following: blood-tonifying herbs: prepared rehmannia root, angelica root, jujube, white peony root, and fleeceflower root; and / or at least one of the following: Yin-tonifying herbs: dendrobium, northern ginseng, southern ginseng, ophiopogon root, asparagus root, polygonatum rhizome, and polygonatum rhizome; and / or at least one of the following: Yang-tonifying herbs: deer antler, epimedium, eucommia bark, dipsacus root, dodder seed, cistanche, psoralea fruit, and alpinia oxyphylla.

[0011] A second aspect of the present invention relates to a method for preparing the above-mentioned traditional Chinese medicine composition, comprising the following steps:

[0012] (1) Weigh the remaining raw materials except for tortoise shell glue, deer antler glue and donkey-hide gelatin, add water and reflux to extract, filter, combine the filtrates, concentrate and obtain the decoction for later use.

[0013] (2) Crush the tortoise shell glue, deer antler glue and donkey-hide gelatin, add them to the decoction in step (1) to dissolve, let cool, and obtain a mixed liquid;

[0014] (3) Add probiotic powder to sterile saline to prepare bacterial suspension, add bacterial suspension to the mixture in step (2), carry out fermentation culture, sterilize, and obtain the Chinese medicine composition.

[0015] Preferably, the reflux extraction in step (1) is performed 2-3 times, with each extraction lasting 1-2 hours, and the amount of water added is 2-5 times the total mass of the extracted raw material. The concentration is performed until the relative density at 60°C is 1.1-1.3 g / mL.

[0016] Preferably, the cooling temperature in step (2) is 36-38℃; the fermentation temperature in step (3) is 36-39℃, and the fermentation time is 36-48h.

[0017] Preferably, the concentration of the bacterial suspension in step (3) is 1-2 mg / mL, and the mass of the bacterial suspension is 1%-2% of the total mass of the raw materials of the traditional Chinese medicine composition.

[0018] Preferably, the preparation method further includes drying the traditional Chinese medicine composition obtained in step (3) to make a dry paste for use in the preparation of solid dosage forms.

[0019] More preferably, the drying includes spray drying, vacuum drying, or conventional drying.

[0020] A third aspect of the present invention relates to a formulation comprising the above-described traditional Chinese medicine composition or a traditional Chinese medicine composition prepared according to the above-described preparation method.

[0021] Preferably, the formulation further includes pharmaceutically acceptable excipients.

[0022] Preferably, the dosage form of the preparation is selected from oral liquid, granules, pills, capsules or tablets.

[0023] The fourth aspect of the present invention relates to the use of the above-mentioned traditional Chinese medicine composition or the traditional Chinese medicine composition or preparation obtained by the above-mentioned preparation method in the preparation of products for treating symptoms of deficiency of qi, blood, yin and yang such as fatigue, myocardial ischemia, cerebral hypoxia, and kidney essence deficiency, and / or for treating and preventing leukopenia caused by bone marrow suppression and / or DNA reduction after radiation and / or low immunity, high altitude hypoxia, and sudden cardiac death.

[0024] Preferably, the deficiency syndrome of Qi, blood, Yin and Yang includes congenital insufficiency or acquired malnutrition (such as serious illness, chronic illness, overwork, excessive sexual activity, etc.) leading to severe depletion of essence and blood, as well as symptoms such as aversion to cold and cold limbs (Yang deficiency), five-center heat, tidal fever and night sweats (Yin deficiency).

[0025] The beneficial effects of this invention are: the fermented product obtained by optimizing the raw materials and fermenting the drug raw materials with probiotics has a better effect than the comparative proportions in treating symptoms of deficiency of Qi, blood, Yin and Yang such as fatigue, myocardial ischemia, cerebral hypoxia, and kidney essence deficiency, and / or treating and preventing leukopenia caused by bone marrow suppression and / or DNA reduction after radiation and / or low immunity, high altitude hypoxia, and prevention of sudden cardiac death. Attached Figure Description

[0026] Figure 1 HPLC chromatograms of rare ginsenoside Rg3 in the traditional Chinese medicine compositions obtained in Example 3 and Comparative Example 5, where A is the ginsenoside Rg3 standard, B is Comparative Example 5, and C is Example 3. Detailed Implementation

[0027] The present invention will be further described below with reference to specific embodiments, and the advantages and features of the present invention will become clearer as a result. However, these embodiments are merely exemplary and do not constitute any limitation on the scope of the present invention. Those skilled in the art should understand that modifications or substitutions can be made to the details and form of the technical solutions of the present invention without departing from the spirit and scope of the present invention, but all such modifications and substitutions fall within the protection scope of the present invention.

[0028] Example 1

[0029] A traditional Chinese medicine composition for replenishing qi, blood, yin and yang, wherein the raw materials of the traditional Chinese medicine composition by weight are: 1 part tortoise shell glue, 2 parts deer antler glue, 1 part donkey-hide gelatin, 2 parts ginseng, 2 parts astragalus, 6 parts codonopsis and 2 parts wolfberry, which are prepared by fermentation of the aforementioned raw materials by a mixture of Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus casei and Lactobacillus rhamnosus.

[0030] The specific preparation method is as follows:

[0031] (1) Weigh the amount of ginseng, astragalus, codonopsis and wolfberry in the formula, add 5 times the amount of water and reflux extract twice, 2 hours each time. Combine the filtrates and concentrate to a relative density of 1.2 g / mL at 60℃ to obtain the decoction for later use.

[0032] (2) Crush the tortoise shell glue, deer antler glue and donkey-hide gelatin, add them to the decoction in step (1) and melt them, then cool them to 36°C to obtain a mixed liquid for later use;

[0033] (3) Add equal amounts of Lactobacillus plantarum powder, Lactobacillus acidophilus powder, Lactobacillus casei powder and Lactobacillus rhamnosus powder to sterile physiological saline to prepare a bacterial suspension of 1 mg / mL. Add the bacterial suspension to the mixture in step (2). The mass of the bacterial suspension is 1% of the total mass of the raw materials of the traditional Chinese medicine composition. Ferment and culture the mixture at 36°C for 48 hours. Sterilize the mixture to obtain the traditional Chinese medicine composition.

[0034] Example 2

[0035] A traditional Chinese medicine composition for replenishing qi, blood, yin and yang, wherein the raw materials of the traditional Chinese medicine composition by weight are: 2 parts tortoise shell glue, 1 part deer antler glue, 2 parts donkey-hide gelatin, 5 parts ginseng, 10 parts astragalus, 4 parts codonopsis and 12 parts wolfberry, which are prepared by fermentation of the aforementioned raw materials with a mixture of Lactobacillus plantarum, Lactobacillus acidophilus, Bacillus subtilis and Lactobacillus bulgaricus.

[0036] The specific preparation method is as follows:

[0037] (1) Weigh the amount of ginseng, astragalus, codonopsis and wolfberry in the formula, add 5 times the amount of water and reflux extract 3 times, 2h each time, combine the filtrates and concentrate to a relative density of 1.1g / mL at 60℃ to obtain the decoction for later use.

[0038] (2) Crush the tortoise shell glue, deer antler glue and donkey-hide gelatin in the formula, add them to the decoction in step (1) to dissolve, and then cool to 38°C to obtain a mixed liquid for later use.

[0039] (3) Mix equal amounts of Lactobacillus plantarum powder, Lactobacillus acidophilus powder, Bacillus subtilis powder and Lactobacillus bulgaricus powder and add them to sterile physiological saline to prepare a bacterial suspension of 2 mg / mL. Add the bacterial suspension to the mixture in step (2). The mass of the bacterial suspension is 1% of the total mass of the raw materials of the traditional Chinese medicine composition. Ferment and culture the mixture at 39°C for 36 hours. Sterilize the mixture to obtain the traditional Chinese medicine composition.

[0040] Example 3

[0041] A traditional Chinese medicine composition for replenishing qi, blood, yin and yang, wherein the raw materials of the traditional Chinese medicine composition by weight are: 1 part tortoise shell glue, 1 part deer antler glue, 1 part donkey-hide gelatin, 3 parts ginseng, 3 parts astragalus, 3 parts codonopsis and 4 parts wolfberry, which are prepared by fermentation of the aforementioned raw materials by a mixture of Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus casei and Lactobacillus rhamnosus.

[0042] The specific preparation method is as follows:

[0043] (1) Weigh the amount of ginseng, astragalus, codonopsis and wolfberry in the formula, add 5 times the amount of water and reflux extract 3 times, 1.5h each time, combine the filtrates and concentrate to a relative density of 1.3g / mL at 60℃ to obtain the decoction for later use.

[0044] (2) Crush the tortoise shell glue, deer antler glue and donkey-hide gelatin in the formula, add them to the decoction in step (1) to dissolve, and then cool to 37°C to obtain a mixed liquid for later use.

[0045] (3) Mix Lactobacillus plantarum powder, Lactobacillus acidophilus powder, Lactobacillus casei powder and Lactobacillus rhamnosus powder and add them to sterile physiological saline to prepare a bacterial suspension of 1 mg / mL. Add the bacterial suspension to the mixture in step (2). The mass of the bacterial suspension is 1% of the total mass of the raw materials of the traditional Chinese medicine composition. Ferment and culture the mixture at 38°C for 40 hours. Sterilize the mixture to obtain the traditional Chinese medicine composition.

[0046] Comparative Example 1

[0047] The difference from Example 3 is that the mixed bacterial powder does not contain Lactobacillus plantarum. Everything else is the same as Example 3.

[0048] Comparative Example 2

[0049] The difference from Example 3 is that the mixed bacterial powder does not contain Lactobacillus acidophilus. Everything else is the same as Example 3.

[0050] Comparative Example 3

[0051] The difference from Example 3 is that the mixed bacterial powder does not contain Lactobacillus rhamnosus. Everything else is the same as Example 3.

[0052] Comparative Example 4

[0053] The difference from Example 3 is that the mixed bacterial powder does not contain Lactobacillus casei. Everything else is the same as Example 3.

[0054] Comparative Example 5

[0055] The difference from Example 3 is that no fermentation treatment is performed, i.e., the preparation method does not include step (3). Everything else is the same as Example 3.

[0056] Comparative Example 6

[0057] I purchased Guilu Erxian Ointment from Tongrentang Pharmacy.

[0058] Experimental Example 1: Detection of Small Molecule Peptides in a Composition

[0059] Methods: High performance size exclusion chromatography was used.

[0060] High-performance liquid chromatography (HPLC): Equipped with a UV detector and a chromatography workstation containing GPC data processing software. Column: TSK gel G2000SWXL 300mm × 7.8mm (GELLOT502R). Mobile phase: acetonitrile-water-trifluoroacetic acid, volume ratio 40:60:0.05; detection wavelength: 220nm; flow rate: 0.5mL / min; column temperature: 30℃; injection volume: 10μL. To ensure the chromatographic system meets the detection requirements, under the above chromatographic conditions, the column efficiency (theoretical plate number N) of the gel chromatography column, calculated based on the tripeptide standard (acetaminophen-acetaminophen-acetaminophen) peak, should not be less than 5000, and the partition coefficient (Kd) of the protein peptide should be between 0 and 1.

[0061] Preparation of relative molecular mass calibration curves: Peptide standard solutions with concentrations of 1.0 g / L above were prepared using the mobile phase. After filtration through a 0.2 μm polytetrafluoroethylene (PTFE) filter membrane, the solutions were injected separately to obtain chromatograms of the series of standards. The relative molecular mass calibration curves and their equations were obtained by plotting the logarithm of the relative molecular mass (IgMW) against retention time or by performing linear regression.

[0062] Sample preparation: Weigh 125.0 mg of sample using weighing paper, transfer it to a 25 mL volumetric flask, dilute to the mark with mobile phase, sonicate for 10 min to fully dissolve and mix the sample, filter through a 0.2 μm polytetrafluoroethylene filter membrane, and use the filtrate for determination.

[0063] Calculation of relative molecular mass: The prepared sample solution is injected and analyzed under chromatographic conditions. Then, using GPC data processing software, the chromatogram and data of the sample are calculated based on the relative molecular mass correction curve equation to obtain the relative molecular mass and distribution range of collagen peptides in the sample. The sum of the relative percentages of collagen peptides with a relative molecular mass less than 10,000 is calculated using the peak area normalization method.

[0064] The test results are shown in Table 1 below.

[0065] Table 1: Total Nitrogen Content of the Composition

[0066]

[0067] As shown in Table 1 above, the content of small molecule peptides increased to varying degrees under the action of different probiotic compositions. Among them, the fermentation composition prepared in Example 3 had the highest content of dipeptides and tripeptides. In Comparative Examples 1-4, after removing one of the four strains during fermentation, the content of small molecule peptides increased compared to the unfermented group, but was significantly lower than that of the Example group. This indicates that fermentation treatment under specific strain composition is beneficial to the decomposition and cleavage of peptides, forming small molecule peptides that are more easily utilized and absorbed. The dipeptide content in Comparative Example 5 (without fermentation treatment) and Comparative Example 6 (existing commercial products) was significantly lower than that of this application, and the tripeptide content in Comparative Example 5 (without fermentation treatment) and Comparative Example 6 (existing commercial products) was also lower than that of this application.

[0068] Experimental Example 2: Detection of Rare Ginsenosides in the Composition

[0069] Method: High performance liquid chromatography (HPLC) was used.

[0070] Waters e2695 high-performance liquid chromatograph, 2998 PDA UV detector. Mobile phase: Column: Welch XB-C18 (4.6 mm × 250 mm, 5 μm); Column temperature: 30℃; Detection wavelength: 203 nm; Flow rate: 1 mL / min; Injection volume: 20 µL; Mobile phase: acetonitrile (B) - 0.1% phosphoric acid aqueous solution (D) gradient elution, elution program: 0-20 min, 20%-20% B; 20-30 min, 20%-35% B; 30-65 min, 35%-60% B; 65 min-75 min, 60%-20% B; 75-85 min, 20%-20% B. See chromatograms below. Figure 1 The content results are shown in Table 2.

[0071] Standard curve of ginsenoside Rg3: y = 349198x - 84689, R 2 =0.9999.

[0072] Table 2: Content of Ginsenoside Rg3 in the Composition

[0073]

[0074] The results of the determination of the content of rare ginsenoside Rg3 in ginseng showed that the content of rare ginsenoside in the fermented composition was significantly higher than that in the unfermented composition. At the same time, the content of ginsenoside Rg3 in the examples with the specific probiotic composition of the present invention was significantly higher than that in comparative examples 1-4.

[0075] Experimental Example 3: Effect of a Traditional Chinese Medicine Composition on Hemoglobin and White Blood Cell Elevation in a Cyclophosphamide-Induced Bone Marrow Suppression Rats

[0076] Experimental animals: 130 male SD rats, weighing 220±20g.

[0077] Methods: Animals were randomly assigned to groups of 10 each, including a blank control group, a model group, a positive control group, Example 1-Example 2 groups, Example 3 low-dose group, Example 3 medium-dose group, Example 3 high-dose group, and Comparative Examples 1-5 groups. The positive control group received commercially available Guilu Erxian Ointment (Comparative Example 6). Except for the blank control group, all other groups underwent tail bleeding starting on day 1, 1 mL / animal, every other day for a total of 5 bleeding sessions. On day 2, cyclophosphamide 40 mg / kg was injected intraperitoneally, every other day for a total of 4 injections, with the last three injections at a dose of 20 mg / kg.

[0078] Administration method: The blank group and the model group were administered an equal volume of distilled water by gavage daily; the drug administration group was administered the drug by gavage once daily starting from the first day of the experiment, with the following dosages: positive control group: 2.970 g / kg, Examples 1-2 and Comparative Examples 1-5: 2.970 g crude drug / kg, Example 3 high-dose group: 5.940 g crude drug / kg, Example 3 medium-dose group: 2.970 g crude drug / kg, Example 3 low-dose group: 1.485 g crude drug / kg, with an administration volume of 1 mL / 200 g bw, and all were administered for 21 days.

[0079] After administering the medication as described above, blood was drawn from the orbital cavity on day 21 for routine blood analysis. The experimental results are shown in Table 3.

[0080] Table 3: Effects of Traditional Chinese Medicine Combinations on a Rat Model of Myelosuppression

[0081]

[0082] Note: Compared with the blank group, # P<0.05, ## P<0.01; compared with the model group, P<0.05, P<0.01; compared with the dosage group in Example 3, & P<0.05.

[0083] According to the data from the rat qi and blood deficiency experiment in Table 3 above, the combined fermentation product prepared in the embodiments of the present invention can significantly improve various blood count indicators. The results of the immune function test show that it can significantly improve the blood count indicators of erythrocytes (RBC), white blood cells (WBC), and hemoglobin (HGB) in rats with qi and blood deficiency induced by cyclophosphamide, and significantly increase white blood cell count. All of these indicators are significantly better than those in the model group. This further verifies that the combined fermentation product claimed in this invention can enhance erythroid hematopoietic function, thereby achieving the effect of simultaneously replenishing qi and blood. The functions of the combined fermentation products with different substrate ratios and different bacterial strains varied in the treatment groups, with the blood count indicators in Example 3 showing the most significant improvement.

[0084] Conclusion: The combined fermentation product has the effects of replenishing qi and blood, improving bone marrow suppression, and increasing white blood cells and hemoglobin, and it does not cause stagnation or impair spleen function.

[0085] Experiment Example 4: Study on Radiation Resistance

[0086] From the perspective of traditional Chinese medicine theory, exogenous "ionizing radiation" is a strong external pathogenic factor, characterized by its malevolent nature and belonging to the categories of "toxic fire" and "violent qi." It rapidly depletes the body's vital energy, leading to a severe imbalance of yin and yang. A cobalt-irradiated mouse survival rate and pharmacodynamic experiment was conducted to examine the survival rate of irradiated mice under drug administration / without drug administration and to compare the efficacy between different groups.

[0087] Laboratory animals: Clean-grade ICR mice, male, weighing 18-20g.

[0088] Drugs: Traditional Chinese medicine compositions of Examples 1-3 and Comparative Examples 1-6; Positive agent: Black fungus polysaccharide.

[0089] Methods and Administration: Mice were randomly assigned to four groups: a blank control group, a model group, a positive control group, groups 1-2 of the examples, groups 1-6 of the comparative examples, a high-dose group of the example 3, and a low-dose group of the example 3, with eight mice in each group. The blank control group and the model group received an equal volume of physiological saline via gavage daily. The positive control group received 800 mg / kg of black fungus polysaccharide, the groups 1-2 and 1-6 received 800 mg / kg, the high-dose group of the example 3 received 800 mg / kg, and the low-dose group of the example 3 received 20 mg / kg. All medications were administered orally via gavage once daily for 30 consecutive days. On day 31, all groups except the blank control group received [treatment / treatment]. 60 Coy-ray irradiation was administered for a total absorbed dose of 4.0 Gy at an absorbed dose rate of 0.5 Gy / min, with a skin source irradiation distance of 80 cm. The irradiation lasted for 14 days.

[0090] Experimental environmental control conditions: The experimental site was well-ventilated and protected from light; room temperature 23±1℃; humidity 50±5%; 12 / 12h light and dark alternation. Animals were acclimatized for 5-7 days before the experiment. Mice had free access to water and food during the experiment.

[0091] The number of mice surviving on day 1 and day 14 of irradiation was recorded. The survival rate of mice after irradiation is shown in Table 4.

[0092] After the experiment, mice were euthanized by cervical dislocation, and the femur was dissected. All bone marrow cells in the femur were flushed out using 1 mL of D-Hank's solution in a syringe. The cell suspension was then thoroughly dispersed in the suspension using a syringe with a No. 4 needle. DNA content was measured at 260 nm using a spectrophotometer. The results are shown in Table 5.

[0093] Table 4: Effects on survival of irradiated mice

[0094]

[0095] Note: Compared with before administration, P<0.05, P<0.01.

[0096] The experimental data in Table 3 above show that the combined fermentation product prepared in the embodiments of the present invention can increase the survival rate of irradiated mice. Compared with the irradiated group, each embodiment group has a significant effect on the survival rate of irradiated mice (P<0.05). Among them, the high-dose group in Example 3 is more effective than the positive drug group. This indicates that the combined fermentation product has a certain protective effect against radiation.

[0097] Table 5: Effects on DNA content in bone marrow of irradiated mice (mean ± standard deviation)

[0098]

[0099] Note: Compared with the blank group, ## P<0.01; compared with the model group, each drug-treated group P<0.05, P<0.01, P<0.001; compared with the positive control group, each treatment group & P<0.05, && P<0.01.

[0100] The experimental data in Table 5 above show that the combined fermentation product prepared in the embodiments of the present invention can increase the content of bone marrow DNA in irradiated mice. Compared with the irradiated group, each embodiment group has a significant effect on bone marrow DNA in irradiated mice (P<0.05). Among them, the high-dose group in Example 3 has a better effect compared with the positive drug group. This indicates that the combined fermentation product has a certain protective effect against radiation.

[0101] Conclusion: The combined fermentation product can improve severe bone marrow depletion after radiation.

[0102] The combined fermentation product can improve severe bone marrow depletion and improve survival rate after radiation, indicating that the combined fermentation product can replenish the body's Yin and Yang.

[0103] Experimental Example 5: Clinical Application

[0104] Inclusion criteria: 1) Acceptance of post-medication blood count measurements. 2) Meeting the following symptoms of Yin and Yang deficiency: lower back and knee weakness, five-center heat (palms, soles, and chest), afternoon fever, bone steaming fever, facial flushing, dry mouth and tongue, constipation, night sweats, aversion to cold, cold extremities, preference for warmth and pressure, fatigue, etc. 3) Chemotherapy and radiotherapy patients. 4) Surgical patients with blood loss. 5) Elderly, frail individuals, and those with weakened immunity. 6) Individuals with other chronic diseases accompanied by Qi and blood deficiency.

[0105] Criteria for judgment:

[0106] Significant effect: Blood counts returned to normal; symptoms of Yin-Yang deficiency and Qi-Blood deficiency were significantly improved;

[0107] Effective: Blood counts tend to be normal, or at least one of the three indicators (RBC, WBC, HGB) is improved compared to before medication; symptoms of qi and blood deficiency are improved;

[0108] Ineffective: Blood counts indicate blood and qi deficiency; symptoms show no improvement.

[0109] Dosage regimen: Take the herbal composition prepared in Example 3 twice daily, on an empty stomach in the morning and evening, 30-50 mL each time. After 21-28 days of continuous treatment, blood tests should be performed, and the improvement of symptoms should be recorded.

[0110] Results: The statistical results are shown in Table 6.

[0111] Table 6: Clinical Efficacy Trials

[0112]

[0113] Two patients who did not respond to the medication were elderly individuals with long-term chronic underlying diseases. They were also taking other medications concurrently during treatment, making it difficult to rule out interference from these other drugs; however, their fatigue improved. Chemotherapy and radiotherapy patients showed significant effects, with most experiencing effectiveness and a few achieving marked effectiveness. Patients experiencing lower back and knee pain experienced significant recovery in strength and a marked disappearance of pain and soreness after taking the medication.

[0114] Conclusion: The overall effective rate for patients with deficiency of essence and blood, characterized by soreness and weakness of the lower back and knees, cold limbs, hot flashes and night sweats, was 91.52%. It can improve deficiency symptoms, increase blood count indicators, and has the effect of increasing white blood cell count and red blood cell count.

[0115] In summary, the traditional Chinese medicine composition provided by this invention has the effect of simultaneously replenishing Yin and Yang, Qi and Blood. It is used to replenish the deficiency of essence and blood, and to treat deficiency of Yin and Yang, deficiency of Qi and Blood, soreness and weakness of waist and knees, weakness of body, low immunity, and rapid increase of hemoglobin and white blood cells, as well as the effects of chemotherapy, radiotherapy, surgery, old age and weakness, postpartum, and chronic illness.

[0116] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, and is not intended to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions made by those skilled in the art to the technical solution of the present invention do not depart from the essence and scope of the technical solution of the present invention.

Claims

1. A traditional Chinese medicine composition for tonifying qi, blood, yin, and yang, characterized in that, The traditional Chinese medicine composition is made from tortoise shell glue, deer antler glue, donkey-hide gelatin, ginseng, astragalus, codonopsis, and wolfberry. The traditional Chinese medicine composition is prepared by fermentation. The mass ratio of tortoise shell glue, deer antler glue, donkey-hide gelatin, ginseng, astragalus, codonopsis, and wolfberry is 1:0.5-2:0.5-2:1-4:2-5:2-6:2-6. The fermentation process uses four types of probiotics in a mass ratio of 1:1:1:

1. The four probiotics consist of Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus casei, and Lactobacillus rhamnosus, or Lactobacillus plantarum, Lactobacillus acidophilus, Bacillus subtilis, and Lactobacillus bulgaricus. The fermentation process involves preparing the probiotic powder into a bacterial suspension with a concentration of 1-2 mg / mL; the mass of the bacterial suspension is 1%-2% of the total mass of the raw materials in the traditional Chinese medicine composition.

2. A method for preparing the traditional Chinese medicine composition according to claim 1, characterized in that, Includes the following steps: (1) Weigh the remaining raw materials except for tortoise shell glue, deer antler glue and donkey-hide gelatin according to the formula, add water and reflux to extract, filter, combine the filtrates, concentrate to obtain decoction; (2) Crush the tortoise shell glue, deer antler glue and donkey-hide gelatin, add them to the decoction in step (1) to dissolve, let cool, and obtain a mixed liquid; (3) Add probiotic powder to sterile saline to prepare bacterial suspension, add bacterial suspension to the mixture in step (2), ferment and culture, sterilize, and the traditional Chinese medicine composition is obtained.

3. The preparation method according to claim 2, characterized in that, The preparation method further includes drying the traditional Chinese medicine composition obtained in step (3) to make a dry paste for use in the preparation of solid dosage forms; wherein the drying includes spray drying, vacuum drying or conventional drying.

4. A formulation, characterized in that, The preparation contains the traditional Chinese medicine composition according to claim 1 or the traditional Chinese medicine composition prepared by the preparation method according to any one of claims 2-3.

5. The use of a traditional Chinese medicine composition according to claim 1, or a traditional Chinese medicine composition prepared by any one of claims 2-3, or a preparation according to claim 4, in the preparation of a drug for treating fatigue, myocardial ischemia, cerebral hypoxia, deficiency of qi, blood, yin and yang caused by kidney essence deficiency and / or for treating and preventing leukopenia and / or low immunity caused by bone marrow suppression.

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