Radix fici simplicissimae deficiency-tonifying granules and preparation method thereof

Abstract

The invention discloses radix fici simplicissimae deficiency-tonifying granules and a preparation method thereof, and belongs to the technical field of traditional Chinese medicines. The radix fici simplicissimae deficiency-tonifying granules are prepared from the following raw materials in parts by weight: 12-20 parts of radix fici simplicissimae, 7-15 parts of radix codonopsis, 5-10 parts of rhizoma atractylodis macrocephalae, 5-10 parts of radix angelicae sinensis, 4-10 parts of pericarpium citri reticulatae, 3-8 parts of bupleurum chinense, 3-8 parts of rhizoma cimicifugae, 5-10 parts of fructus lycii, 5-10 parts of semen cuscutae, 5-10 parts of fructus psoraleae, 5-10 parts of herba epimedii and 4-10 parts of radix glycyrrhizae preparata. The product can tonify qi, invigorate the spleen, regulate and tonify the liver and the kidney, tonify the middle warmer and raise yang. The traditional Chinese medicine is used for patients with weakness caused by deficiency of spleen and stomach qi, deficiency of liver and kidney, shortness of breath and weakness, asthma, body fever and sweating, less complexion, headache and aversion to cold, soreness and weakness of waist and knees and fatigue syndromes. And chronic diarrhea, rectocele, uterine prolapse and the like caused by sinking of middle-jiao qi.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicines and relates to five-fingered ginseng deficiency-tonifying granules and a preparation method thereof. Background Art

[0002] Traditional Chinese Medicine (TCM) believes that "irregular diet, irregular daily routines, emotional distress, and excessive work and rest" are among the major causes of sub-health. Exogenous pathogens such as the six pathogenic factors and toxins also contribute. "Excessive eating and drinking damages the stomach and intestines," as stated in "Suwen (Suwen) Bilun," and "Improper diet can harm a person in an instant." Irregular and uncontrolled eating can affect gastrointestinal function and harm the body. "Prolonged sitting damages the flesh, prolonged standing damages the bones, prolonged lying damages the qi, and prolonged walking damages the tendons." Irregular daily routines and prolonged lying deplete the body's qi and blood, while an imbalance between work and rest can damage tendons, bones, blood, and flesh.

[0003] "Suwen Shi Congrong Lun" states: "Liver deficiency, spleen deficiency, and kidney deficiency all cause weight gain and discomfort." Weight loss, coupled with fatigue and weakness, is a manifestation of fatigue syndrome in a sub-healthy state. "Wu Chang Zheng Da Lun" states: "Where Yin essence is maintained, a person lives long; where Yang essence descends, a person is born." Yin essence is maintained, meaning when the spleen and stomach are in harmony, the mustard qi rises, and the spring and summer orders are in motion, resulting in longevity; Yang essence descends, meaning when the spleen and stomach are in disharmony, the grain qi flows downward, and the storage order is in motion, resulting in premature death. "Yi Zong Must Read" states: "Once this body exists, it must be nourished by grain qi. Grain enters the stomach, circulates in the six bowels, and qi arrives. It harmonizes the five internal organs, and blood is produced. This is the source of life for the human body, so it is said that the foundation of acquired constitution lies in the spleen." It can be seen that if the spleen functions normally, the source of Qi and blood will be sufficient, the internal organs will function normally, the muscles will be strong and powerful, the limbs will be flexible, the mind will be responsive, the mind will be focused, and the sleep will be good; otherwise, the source of Qi and blood will be insufficient, the internal organs, meridians, limbs, and muscles will be malnourished, and sub-health symptoms such as limb inactivity, muscle atrophy, physical fatigue, and drowsiness will appear.

[0004] With the increasing attention and emphasis on sub-health prevention and treatment, the development and utilization of in-hospital medications for sub-health-related conditions is urgently needed. Compared to Western medicine, traditional Chinese medicine is more widely accepted by patients due to its safety, effectiveness, and minimal side effects.

[0005] Therefore, how to develop a five-fingered ginseng tonic granule and its preparation method for treating sub-health conditioning, spleen and stomach qi deficiency, liver and kidney deficiency and sinking of qi in the middle, which has the effects of invigorating qi and strengthening spleen, regulating liver and kidney, is a technical problem that technicians in this field urgently need to solve. Summary of the Invention

[0006] In view of this, the present invention provides a five-fingered ginseng nourishing granule and a preparation method thereof.

[0007] In order to achieve the above object, the present invention adopts the following technical solutions:

[0008] A five-fingered ginseng deficiency-tonifying granule is prepared by including the following raw materials in parts by weight:

[0009] 12-20 parts of five-fingered peach (Xisi), 7-15 parts of Codonopsis pilosula, 5-10 parts of Atractylodes macrocephala, 5-10 parts of Chinese angelica, 4-10 parts of dried tangerine peel (Citrus tangerine), 3-8 parts of Bupleurum chinense, 3-8 parts of Cimicifuga heracleifolia, 5-10 parts of wolfberry fruit, 5-10 parts of Cuscuta seed (Dodder seed), 5-10 parts of Psoralea corylifolia, 5-10 parts of Epimedium, and 4-10 parts of roasted liquorice.

[0010] The present invention also provides a preparation method of the five-fingered peach nourishing granules, comprising the following steps:

[0011] (1) Weigh the raw materials according to the Wuzhimaotao Buxu Granules;

[0012] (2) soaking the weighed raw materials in water, decocting them twice to obtain a medicinal solution, and filtering the medicinal solution to obtain a filtrate;

[0013] (3) concentrating the obtained filtrate for the first time to obtain a concentrated solution, and concentrating the concentrated solution again to obtain a thick paste of Wuzhimaotao Buxu Granules;

[0014] (4) adding the obtained Wuzhimaotao Buxu Granules paste to the dextrin while stirring, continuing to stir evenly, drying, and crushing to obtain a fine powder;

[0015] (5) The obtained fine powder is stirred evenly, and 85% ethanol is added to moisten and stir to prepare a soft material, and the soft material is sieved and granulated to obtain wet granules;

[0016] (6) drying the obtained wet granules, re-granulating the granules, and stirring the granules to obtain the five-fingered ginseng nourishing granules.

[0017] Furthermore, in step (2), the raw material is soaked in water for 30 minutes and then decocted twice, with 10 times the amount of water added for the first time and 8 times the amount of water added for the second time. Each decoction is for 1 hour. During the decoction process, the steam pressure is controlled at 0.1-0.25 MPa before the medicinal liquid boils, and the steam pressure is controlled at 0.05-0.10 MPa after the medicinal liquid boils, so that the medicinal liquid keeps boiling. After the extraction is completed, the medicinal liquid is filtered with a 100-mesh filter.

[0018] Furthermore, in step (3), the first concentration temperature is 75-95°C, the vacuum degree is -0.01 to -0.09 MPa, and the concentrate is concentrated to a concentrated liquid with a relative density of 1.05-1.15 at 60°C; and the concentrate is concentrated again to a thick paste with a relative density of 1.28-1.35 at 60°C.

[0019] Furthermore, in step (4), the stirring speed is 24 rpm, the stirring time is continued for 5-10 min, the mass ratio of the five-fingered ginseng tonic granule paste and dextrin is 1:(0.8-1.0), the drying temperature is 65-85°C, and the mixture is dried to a moisture content of less than 9.0%, and then crushed to a fine powder and passed through an 80-mesh sieve.

[0020] Furthermore, in step (5), the obtained fine powder is stirred evenly at a stirring speed of 24 rpm for 5-10 min, 85% ethanol is added to moisten and stir to make a soft material until it can be grasped into a ball and falls apart when pressed lightly, and then granulated through a 10-mesh sieve.

[0021] Furthermore, in step (6), the drying temperature is 65-85° C., and the mixture is dried until the moisture content is less than 4.0%. The mixture is sieved with 14-mesh and 60-mesh sieves, and particles that pass through 14-mesh but not 60-mesh are considered qualified particles. The stirring speed is 24 rpm, and the stirring time is 20 minutes.

[0022] The beneficial effects of the present invention are:

[0023] (1) Functions and indications:

[0024] TCM Description: Invigorates Qi and strengthens the spleen, regulates the liver and kidneys, and replenishes the middle and raises Yang. It is used for patients with weak constitutions due to spleen and stomach Qi deficiency, liver and kidney insufficiency, shortness of breath, fatigue, frequent wheezing, fever with sweating, pale complexion, headache, chills, soreness of the waist and knees, and fatigue syndrome with the above symptoms; and for chronic diarrhea, rectal prolapse, and uterine prolapse caused by sinking of the middle Qi.

[0025] Zhuang medicine describes it as tonifying Qi and strengthening the spleen, elevating Yang and strengthening the foundation, nourishing the liver and kidneys, regulating the three channels and two pathways, and enhancing disease resistance. It is used to treat abdominal distension, poor appetite, loose stools, fatigue, shortness of breath, and organ ptosis caused by spleen deficiency and dampness, and the sinking of Qi in the middle; sallow or pale complexion, palpitations, and insomnia caused by Qi and blood deficiency; and arthritis, lumbago, paralysis, numbness of limbs, dizziness, tinnitus, impotence, premature ejaculation, nocturnal emission, enuresis, irregular menstruation, blurred vision, diarrhea, and frequent urination at night caused by liver and kidney deficiency.

[0026] The Chinese medicinal materials in the prescription, including five-fingered ginseng, Codonopsis pilosula, Atractylodes macrocephala, Angelica sinensis, dried tangerine peel, Bupleurum chinense, Cimicifuga heracleifolia, Lycium barbarum, Cuscuta chinensis, Psoralea corylifolia, Epimedium brevicornum, and roasted licorice, are all non-toxic. The prescription composition does not contain the eighteen anti-reactions and nineteen fear incompatibilities. The dosage of five-fingered ginseng, Codonopsis pilosula, Atractylodes macrocephala, Angelica sinensis, dried tangerine peel, Bupleurum chinense, Cimicifuga heracleifolia, Lycium barbarum, Cuscuta chinensis, Psoralea corylifolia, Epimedium brevicornum, and roasted licorice is within the prescribed range and does not exceed the standard.

[0027] (2) Prescription source:

[0028] This prescription, based on the fundamental principle of Traditional Chinese Medicine (TCM) treatment based on "differentiation of symptoms and treatment," draws on the fundamental Zhuang medical theories of "differentiation of symptoms and treatment, the theory of toxicity and pathogenic factors; simplicity of medication, and the importance of specificity." Through repeated and careful research, this formula is formulated to invigorate Qi, strengthen the spleen, and regulate the liver and kidneys. Zhuang medicine theory is centered on the "Three Paths and Two Roads," namely the "Gu Dao" (digestive system), the "Ji Dao" (urinary system), the "Dragon Road" (nervous system), and the "Huo Dao" (circulatory system). Zhuang practitioners believe that human health depends on the patency of these three paths and two roads, as well as the balance of Qi and blood.

[0029] In clinical practice, the inventors discovered that five-fingered ginseng has significant therapeutic effects in tonifying and supporting the weak, and is particularly suitable for patients with sub-health conditions such as frailty, low immunity, and chronic fatigue. Based on this, the inventors organized a professional team to conduct in-depth research on five-fingered ginseng. Combined with the results of modern pharmacological research, they further clarified its pharmacological effects in regulating the body's immune function and improving physical fitness.

[0030] The development of this prescription adhered to the clinical-based, experimental-based, and clinical-focused approach, successfully implementing the "medicine, research, production, and application" principle of transforming research findings in traditional Chinese medicine. This prescription, clinically processed at the Guangxi International Zhuang Medicine Hospital into a decoction, is available to patients. It tonifies Qi and strengthens the spleen, regulates the liver and kidneys, and tonifies the middle and raises Yang. It is indicated for patients with spleen and stomach Qi deficiency, liver and kidney deficiency, and for those experiencing weakness, shortness of breath, fatigue, frequent wheezing, fever and sweating, pale complexion, headache, chills, soreness of the waist and knees, and fatigue syndromes such as these. It also addresses chronic diarrhea, rectal prolapse, and uterine prolapse caused by Qi deficiency. It is effective and can effectively alleviate symptoms.

[0031] In summary, the prescription source of the Wuzhimaotao Buxu Granules of the present invention is the crystallization of the combination of traditional Zhuang and Yao medical theory and modern medical practice, which reflects the hospital's innovation ability and scientific research strength in the fields of traditional Chinese medicine and Zhuang and Yao medicine, and has important clinical application value and promotion significance.

[0032] (3) Theoretical basis:

[0033] This prescription is composed of twelve Chinese medicinal herbs (ethnic medicines), including five-fingered ginseng (Kexisi), Codonopsis pilosula, Atractylodes macrocephala, Angelica sinensis, Tangerine peel (Nenggan), Northern Bupleurum, Cimicifuga heracleifolia, Lycium barbarum, Cuscuta chinensis (Fenqianfa), Psoralea corylifolia, Epimedium (Mengguoyang), and roasted Licorice root. The five-fingered ginseng in the prescription is mild in nature and sweet in taste, and has the effects of strengthening the spleen and replenishing qi, promoting qi and eliminating dampness, relaxing muscles and activating blood circulation. This medicine has a mild nature and taste, replenishes qi without raising qi, strengthens the body without hindering evil, and is not averse even if there is external evil. "Therefore, the prescription uses five-fingered peach to strengthen the spleen and replenish qi, nourish the spleen and benefit the lungs, and is the main medicine. Codonopsis is sweet and flat in nature, and has the effects of strengthening the spleen and benefiting the lungs, nourishing blood and promoting fluid production. It is used for spleen and lung qi deficiency, poor appetite and fatigue, cough and asthma, insufficient qi and blood, sallow complexion, palpitations and shortness of breath, etc. Atractylodes is bitter and sweet in taste, warm in nature, and has the effects of strengthening the spleen and replenishing qi, drying dampness and promoting diuresis, stopping sweating, and calming the fetus. It is used for symptoms such as spleen deficiency, poor appetite, abdominal distension and diarrhea, phlegm and fluid, dizziness and palpitations. Codonopsis and Atractylodes are ministerial medicines that strengthen the spleen and replenish qi, and together they have the effect of tonifying the middle and replenishing qi. Angelica is sweet and spicy in taste, warm in nature, and has the effects of replenishing blood and activating blood circulation, regulating menstruation and relieving pain, and moistening the intestines and promoting bowel movements. In a sub-healthy state, qi deficiency for a long time will inevitably damage the blood, so this prescription is combined with sweet and spicy herbs. The warm angelica nourishes yin and blood. Tangerine peel is bitter and pungent, warm in nature, and has the effects of regulating qi and strengthening the spleen, drying dampness and resolving phlegm. It is used for symptoms of abdominal distension, poor appetite, vomiting and diarrhea, cough and excessive sputum. This prescription is combined with tangerine peel to regulate qi, to help the recovery of ascending and descending, so that the clear and turbid qi can each go its own way, and can regulate qi and stomach, so that all medicines can be nourished without stagnation. Bupleurum is pungent and bitter in taste, and slightly cold in nature. It has the effects of dispersing fever, relieving liver depression, and raising yang. Bupleurum has a variety of pharmacological activities, including anti-inflammatory, immune regulation, promoting bone growth, anti-cancer, and protecting the liver. Cimicifuga is pungent and slightly sweet in taste, and slightly cold in nature. It has the effects of dispelling rashes, clearing away heat and detoxifying, and raising yang. It is clinically used for wind-heat headache, toothache, Mouth ulcers, sore throat and other symptoms. Bupleurum and Cimicifuga lightly lift and disperse it, so as to cooperate with other medicines to help the clear yang rise. Lycium barbarum is sweet in taste and neutral in nature. It has the effects of nourishing the liver and kidney, improving essence and improving eyesight, and has anti-inflammatory, antioxidant, nervous system protection, retinal protection, anti-tumor and other biological activities. Cuscuta chinensis is pungent and sweet in taste, neutral in nature, and has the effects of nourishing the liver and kidney, consolidating essence and reducing urine, calming the fetus, improving eyesight, and stopping diarrhea. It is clinically used for liver and kidney deficiency, soreness of waist and knees, blurred vision and tinnitus, spleen and kidney deficiency diarrhea, etc. Psoralea corylifolia is pungent and bitter in taste, warm in nature, and has the effects of warming the kidney and assisting yang, absorbing qi and relieving asthma, warming the spleen and stopping diarrhea. It is clinically used for kidney yang deficiency, cold pain in waist and knees, and asthma caused by kidney deficiency, etc. Epimedium is pungent and sweet in taste, It is warm and has the effects of tonifying kidney yang, strengthening muscles and bones, and removing rheumatism. It is clinically used for kidney yang deficiency, impotence and spermatorrhea, muscle and bone weakness, rheumatic pain, numbness and cramps, etc. In this prescription, wolfberry fruit and dodder seed nourish the liver and kidney, replenish essence and blood, and psoralea corylifolia and epimedium warm and tonify kidney yang. Angelica sinensis, dried tangerine peel, bupleurum, cimicifuga, wolfberry fruit, dodder seed, psoralea corylifolia, and epimedium are all adjuvants. Licorice is sweet in taste and neutral in nature. It has the effects of tonifying the spleen and replenishing qi, clearing away heat and detoxifying, removing phlegm and relieving cough, relieving acute pain, and harmonizing various medicines. In clinical practice, it has multiple biological functions such as anti-inflammatory, immune regulation, antiviral, anti-tumor, liver protection, and antioxidant. Roasted licorice strengthens the spleen and replenishes qi, and harmonizes various medicines, and is a guiding drug.

[0034] The combination of various medicines can harmonize the spleen and stomach, and enable the essence of water and grain to be produced and transformed efficiently. Then all symptoms of spleen and stomach qi deficiency and liver and kidney deficiency can be cured by themselves.

[0035] In summary, the formula uses Prunus mume as the monarch herb, strengthening the spleen and replenishing qi, tonifying the spleen and benefiting the lungs; Codonopsis pilosula and Atractylodes macrocephala are the assistant herbs, strengthening the spleen and replenishing qi, achieving the function of tonifying the middle and replenishing qi; Angelica sinensis, Tangerine peel, Bupleurum chinense, Cimicifuga heracleifolia, Lycium barbarum fruit, Cuscuta australis, Psoralea corylifolia, and Epimedium are the adjuvant herbs, and Roasted Licorice root is the guiding herb, strengthening the spleen and replenishing qi, and harmonizing the other herbs. The combination of these herbs harmonizes the spleen and stomach, allowing the production of water and grain essence and qi to be activated, thus achieving self-healing of symptoms of spleen and stomach qi deficiency and liver and kidney insufficiency.

[0036] (4) Zhuang medicine prescription principles and explanations:

[0037] The five-fingered ginseng root in the prescription is slightly hot in nature and sweet in taste. It can remove dampness and toxins, replenish qi deficiency and clear the water channels. It is the main medicine and has the main effects of replenishing qi, strengthening the spleen, removing dampness and regulating the three channels. Codonopsis pilosula is neutral in nature and sweet in taste. It can replenish the middle and benefit qi, strengthen the spleen and benefit the lungs. It assists the five-fingered ginseng root in strengthening the qi-replenishing effect, targeting the weakness of the anus, insufficient qi and blood, and improving fatigue, weakness, poor appetite and loose stools. Atractylodes macrocephala is warm in nature and sweet and bitter in taste. It cooperates with the five-fingered ginseng root to remove dampness and resolve phlegm, and is mainly used to treat dampness that obstructs the spleen and urinates. It can relieve abdominal distension and edema caused by liver blood deficiency, and enhance the function of regulating the anus; Angelica sinensis is warm in nature, sweet and spicy in taste, can nourish blood and activate blood circulation, harmonize qi and blood, improve dizziness and irregular menstruation caused by liver blood deficiency, and help the dragon path to be unblocked; Tangerine peel is spicy and warm in nature, bitter in taste, can regulate qi and strengthen spleen, dry dampness and resolve phlegm, and help improve chest and abdominal distension, loss of appetite, vomiting and diarrhea, cough and excessive sputum; Northern Bupleurum is slightly cold in nature, bitter and spicy in taste, can soothe liver and relieve depression, lift yang qi, and lift the sunken middle qi. Improve symptoms such as visceral prolapse and rectal prolapse; Cimicifuga heracleifolia, slightly cold in nature, spicy and sweet in taste, can raise yang and lift the sunken, and when combined with Bupleurum chinense, it can enhance the lifting power; Lycium barbarum, neutral in nature, sweet in taste, nourishes the kidney and nourishes essence, improves symptoms such as soreness of waist and knees, impotence, and blurred vision; Cuscuta chinensis, neutral in nature, spicy and sweet in taste, regulates the anus and replenishes kidney essence, and synergistically treats lumbago, paralysis, impotence, spermatorrhea, enuresis, frequent urination, diarrhea, and frequent urination at night; Psoralea corylifolia, warm in nature, spicy in taste Bitter, warms the kidney qi, stops diarrhea, consolidates essence and reduces urine, improves impotence due to kidney deficiency, spermatorrhea, frequent urination and enuresis, and helps consolidate yang qi; Epimedium is warm in nature, spicy and sweet in taste, dispels wind toxins, removes dampness toxins, replenishes kidney yang, strengthens muscles and bones, and is used to improve muscle and bone weakness, spermatorrhea, arthritis, numbness of limbs, etc., and regulates the dragon path and fire path; Roasted Licorice is neutral in nature, sweet in taste, replenishes the spleen and replenishes qi, harmonizes various medicines, and alleviates the warm and dry nature of various medicines, making the whole prescription warm and nourishing without stagnation, and is a leading medicine. This prescription uses five-fingered ginseng as the main medicine, and is combined with codonopsis pilosula, white atractylodes, etc. to nourish the qi of the spleen and lungs, and strengthen the three channels. Angelica sinensis and tangerine peel harmonize qi and blood, northern bupleurum and cimicifuga lift clear yang, wolfberry, dodder seed, psoralea corylifolia, and epimedium warm and nourish the liver and kidneys as auxiliary medicines, helping the main medicine to improve the main symptoms, and synergistically treat concurrent symptoms, unblocking the two channels, and adding roasted licorice to harmonize the various medicines. The whole prescription is centered on "tonifying qi and strengthening the spleen, raising yang and consolidating the foundation, and nourishing the liver and kidneys", and is suitable for the Zhuang medicine symptoms of "weak anus, blocked dragon channel, and insufficient yin fluid". The whole prescription is used together to nourish qi, blood, essence, spleen, lung, liver, and kidneys, and keep the three channels and two channels unobstructed, so that the disease can be gradually cured.

[0038] This prescription has been used clinically in the form of clinical decoction, decocted in water for oral administration, with one dose taken three times a day, in an appropriate amount each time. In order to facilitate the use, carrying and storage of the medicine for patients, combined with the economic cost of preparation production, energy saving, actual production conditions and better exertion of the efficacy of the medicine, the inventors of the present invention have conducted a project research on this prescription and determined to change the decoction into an oral granule.

[0039] The five-fingered ginseng nourishing granules of the present invention are taken with boiled water, one bag at a time, each bag is 15 grams, and the dose is 3 times a day. BRIEF DESCRIPTION OF THE DRAWINGS

[0040] Figure 1 This is a line graph showing the effect of reserpine on body weight in rats with spleen deficiency.

[0041] Figure 2 This is a bar graph showing the effect of reserpine on the xylose excretion rate in rats with spleen deficiency.

[0042] Figure 3 The bar graphs show the effects of Wuzhimaotao Buxu Granule Qinggao on the secretion of gastrointestinal hormones in rats with spleen deficiency induced by reserpine. (a) shows gastrin (Gas), (b) shows motilin (MTL), (c) shows somatostatin (SS), and (d) shows vasoactive intestinal peptide (VIP).

[0043] Figure 4 The figure is a bar graph showing the effects on gastric emptying and small intestinal propulsion in normal mice, where (a) shows the gastric residual rate and (b) shows the small intestinal propulsion rate.

[0044] Figure 5 The figures are bar graphs showing the effects on gastric emptying and small intestinal propulsion in mice with gastrointestinal motility disorders, where (a) shows the gastric residual rate and (b) shows the small intestinal propulsion rate.

[0045] Figure 6 This is a bar graph showing the effect of adenine on the renal organ coefficient of rats with kidney yang deficiency.

[0046] Figure 7 Figure 3 is a bar graph showing the effects of adenine on serum indicators in rats with kidney yang deficiency. (a) shows the serum creatinine (Cr) content, (b) shows the serum urea nitrogen (UREA) content, (c) shows the serum uric acid (UA) content, (d) shows the serum IgA content, and (e) shows the serum IgG content.

[0047] Figure 8Figure 3 Effects of adenine on renal pathological changes in rats with kidney yang deficiency induced by adenine (HE staining, ×100). (a) is the blank control group, (b) is the model control group, (c) is the high-dose group of Fructus quinquefolii, (d) is the medium-dose group of Fructus quinquefolii, (e) is the low-dose group of Fructus quinquefolii, and (f) is the positive control group.

[0048] Figure 9 The figure is a bar graph showing the effects of adenine on serum indicators of mice with kidney yang deficiency, where (a) shows the testosterone (T) content in serum and (b) shows the cortisol content in serum. DETAILED DESCRIPTION

[0049] The following is a clear and complete description of the technical solutions in the embodiments of the present invention. Obviously, the embodiments described are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making any creative efforts are within the scope of protection of the present invention.

[0050] Quality standards for raw materials (medicinal materials) used in the production of the medicines of the present invention:

[0051] Five-fingered ginseng: See Volume 2 of "Quality Standards of Zhuang Medicine in Guangxi Zhuang Autonomous Region" (2011 edition), pages 47-48.

[0052] Codonopsis pilosula: See pages 293-294 of Volume 1 of the 2020 edition of the Pharmacopoeia of the People's Republic of China.

[0053] Atractylodes: See pages 107-108 of Volume 1 of the 2020 edition of the Pharmacopoeia of the People's Republic of China.

[0054] Angelica: See page 139 of the 2020 edition of the Pharmacopoeia of the People's Republic of China.

[0055] Tangerine peel: see 199-200 of the 2020 edition of the Pharmacopoeia of the People's Republic of China, see Guangxi

[0056] Zhuang Autonomous Region Zhuang Medicine Quality Standards, Part 2 (2011 edition), pages 157-158.

[0057] Bupleurum: See page 293 of Volume 1 of the 2020 edition of the Pharmacopoeia of the People's Republic of China.

[0058] Cimicifuga: See pages 75-76 of Volume 1 of the 2020 edition of the Pharmacopoeia of the People's Republic of China.

[0059] Wolfberry: See pages 260-261 of the 2020 edition of the Pharmacopoeia of the People's Republic of China.

[0060] Cuscuta: See page 322 of Volume 1 of the 2020 edition of the Pharmacopoeia of the People's Republic of China, and pages 259-260 of Volume 2 of the Guangxi Zhuang Autonomous Region Zhuang Medicine Quality Standards (2011 edition).

[0061] Psoralea corylifolia: See page 195 of Volume 1 of the 2020 edition of the Pharmacopoeia of the People's Republic of China.

[0062] Epimedium: See Pharmacopoeia of the People's Republic of China 2020 edition, pp. 340-342, see Guangxi Zhuang

[0063] Quality Standards for Zhuang Medicine in the Autonomous Region of China, Part 2 (2011 edition), pages 278-279.

[0064] Roasted licorice: See pages 88-89 of Volume 1 of the 2020 edition of the Pharmacopoeia of the People's Republic of China.

[0065] Example 1

[0066] The preparation method of Wuzhimaotao nourishing granules comprises the following steps:

[0067] (1) Weigh 15g of Prunus mume, 9g of Codonopsis pilosula, 6g of Atractylodes macrocephala, 6g of Angelica sinensis, 5g of dried tangerine peel, 3g of Bupleurum chinense, 3g of Cimicifuga heracleifolia, 6g of Lycium barbarum, 6g of Cuscuta seed, 6g of Psoralea corylifolia, 6g of Epimedium, and 5g of Radix Glycyrrhizae;

[0068] (2) Soaking the weighed raw materials in water for 30 minutes, decocting them twice, adding 10 times the amount of water for the first time and 8 times the amount of water for the second time, and decocting them for 1 hour each time. During the decocting process, the steam pressure was controlled at 0.2 MPa before the boiling of the medicinal liquid and at 0.08 MPa after the boiling of the medicinal liquid, so that the medicinal liquid was kept boiling. After the extraction was completed, the medicinal liquid was obtained, and the medicinal liquid was filtered with a 100-mesh filter to obtain a filtrate;

[0069] (3) concentrating the obtained filtrate for the first time, wherein the concentration temperature for the first concentration is 80° C. and the vacuum degree is −0.05 MPa, to a concentrate having a relative density of 1.05-1.12 at 60° C., and concentrating the concentrate again to a thick paste of Wuzhimaotao Buxu Granules having a relative density of 1.30 at 60° C.;

[0070] (4) adding the obtained Wuzhi Maotao Buxu Granule Paste to the dextrin while stirring, wherein the mass ratio of the Wuzhi Maotao Buxu Granule Paste to the dextrin is 1:0.9, continuing stirring for 8 minutes at a stirring speed of 24 rpm, drying at a drying temperature of 75° C. until the moisture content is less than 9.0%, and grinding into fine powder and passing through an 80-mesh sieve to obtain a fine powder;

[0071] (5) The obtained fine powder was stirred uniformly at a stirring speed of 24 rpm for 8 min, and 85% ethanol was added to moisten and stir the soft material until it could be lumped together and fell apart when pressed lightly. The soft material was granulated through a 10-mesh sieve to obtain wet granules;

[0072] (6) Dry the obtained wet granules at a drying temperature of 70° C. until the moisture content is less than 4.0%, and sieve the granules with 14-mesh and 60-mesh sieves, stirring them evenly at a stirring speed of 24 rpm for 20 minutes to obtain the Wuzhimaotao tonic granules.

[0073] Example 2

[0074] The preparation method of Wuzhimaotao nourishing granules comprises the following steps:

[0075] (1) Weigh 12g of Prunus mume, 7g of Codonopsis pilosula, 5g of Atractylodes macrocephala, 5g of Angelica sinensis, 4g of dried tangerine peel, 5g of Bupleurum chinense, 5g of Cimicifuga heracleifolia, 5g of Lycium barbarum fruit, 5g of Cuscuta seed, 5g of Psoralea corylifolia, 5g of Epimedium, and 4g of Radix Glycyrrhizae;

[0076] (2) Soaking the weighed raw materials in water for 30 minutes, decocting them twice, adding 10 times the amount of water for the first time and 8 times the amount of water for the second time, and decocting them for 1 hour each time. During the decocting process, the steam pressure was controlled at 0.1 MPa before the boiling of the medicinal liquid and at 0.05 MPa after the boiling of the medicinal liquid, so that the medicinal liquid was kept boiling. After the extraction was completed, the medicinal liquid was obtained, and the medicinal liquid was filtered with a 100-mesh filter to obtain a filtrate;

[0077] (3) The obtained filtrate is concentrated for the first time at a concentration temperature of 75° C. and a vacuum degree of −0.01 MPa to a concentrate having a relative density of 1.05 at 60° C. The concentrate is concentrated again to a thick paste of Wuzhimaotao Buxu Granules having a relative density of 1.28 at 60° C.;

[0078] (4) adding the obtained Wuzhi Maotao Buxu Granule Paste to the dextrin while stirring, wherein the mass ratio of the Wuzhi Maotao Buxu Granule Paste to the dextrin is 1:0.8, continuing stirring for 5 minutes at a stirring speed of 24 rpm, drying at a drying temperature of 65-85° C. until the moisture content is less than 9.0%, and grinding into fine powder and passing through an 80-mesh sieve to obtain a fine powder;

[0079] (5) The obtained fine powder was stirred uniformly at a stirring speed of 24 rpm for 5 min, and 85% ethanol was added to moisten and stir the soft material until it could be lumped together and fell apart when pressed lightly. The soft material was granulated through a 10-mesh sieve to obtain wet granules;

[0080] (6) Dry the obtained wet granules at a drying temperature of 65° C. until the moisture content is less than 4.0%, screen the granules with 14-mesh and 60-mesh sieves, and stir them evenly at a stirring speed of 24 rpm for 20 minutes to obtain the Wuzhimaotao tonic granules.

[0081] Example 3

[0082] The preparation method of Wuzhimaotao nourishing granules comprises the following steps:

[0083] (1) Weigh 20g of Prunus mume, 15g of Codonopsis pilosula, 10g of Atractylodes macrocephala, 10g of Angelica sinensis, 10g of Tangerine peel, 8g of Bupleurum chinense, 8g of Cimicifuga heracleifolia, 10g of Lycium barbarum, 10g of Cuscuta seed, 10g of Psoralea corylifolia, 10g of Epimedium, and 10g of Radix Glycyrrhizae;

[0084] (2) soaking the weighed raw materials in water for 30 minutes, decocting them twice, adding 10 times the amount of water for the first time and 8 times the amount of water for the second time, and decocting them for 1 hour each time. During the decocting process, the steam pressure was controlled at 0.25 MPa before the medicinal liquid boiled, and the steam pressure was controlled at 0.10 MPa after the medicinal liquid boiled, so that the medicinal liquid was kept boiling. After the extraction was completed, the medicinal liquid was obtained, and the medicinal liquid was filtered with a 100-mesh filter to obtain a filtrate;

[0085] (3) The obtained filtrate is concentrated for the first time at a concentration temperature of 95° C. and a vacuum degree of −0.09 MPa to a concentrate having a relative density of 1.15 at 60° C. The concentrate is concentrated again to a thick paste of Wuzhimaotao Buxu Granules having a relative density of 1.35 at 60° C.;

[0086] (4) adding the obtained Wuzhi Maotao Buxu Granule Paste to the dextrin while stirring, wherein the mass ratio of the Wuzhi Maotao Buxu Granule Paste to the dextrin is 1:1.0, continuing stirring for 10 minutes at a stirring speed of 24 rpm, drying at a drying temperature of 85° C. until the moisture content is less than 9.0%, and grinding into fine powder and passing through an 80-mesh sieve to obtain a fine powder;

[0087] (5) The obtained fine powder was stirred uniformly at a stirring speed of 24 rpm for 10 min, and 85% ethanol was added to moisten and stir the soft material until it could be lumped together and fell apart when pressed lightly. The soft material was granulated through a 10-mesh sieve to obtain wet granules;

[0088] (6) Dry the obtained wet granules at 85° C. until the moisture content is less than 4.0%, screen the granules with 14-mesh and 60-mesh sieves, and stir them evenly at a stirring speed of 24 rpm for 20 minutes to obtain the Wuzhimaotao tonic granules.

[0089] The subsequent experiments were carried out using the Wuzhimaotao Buxu Granules of Example 1.

[0090] 1. Inspection

[0091] 1.1 Properties: The properties of three batches of preparations (230901, 230902, and 230903) were observed. The results are shown in Table 1.

[0092] Table 1 Results of the properties of three batches of Wuzhimaotao Buxu Granule samples

[0093] batch number Appearance 230901 Brown-yellow granules; slightly fragrant and slightly sweet 230902 Brown-yellow granules; slightly fragrant and slightly sweet 230903 Brown-yellow granules; slightly fragrant and slightly sweet

[0094] 1.2 Solubility: According to the "Solubility" test under Item 0104 of the 2020 edition of the Chinese Pharmacopoeia, three batches of this product (230901, 230902, and 230903), one bag per batch (15g / bag), were added to 200 mL of hot water and stirred for 5 minutes. Observe immediately. The solution should be completely dissolved or slightly turbid, with no foreign matter or burnt debris. The results are shown in Table 2.

[0095] Table 2 Solubility test results of three batches of Wuzhimaotao Buxu Granules

[0096] batch number result 230901 All melted, no foreign matter, no burnt debris. 230902 All melted, no foreign matter, no burnt debris. 230903 All melted, no foreign matter, no burnt debris.

[0097] 1.3 Extract: Take 3 batches of this product (230901, 230902, 230903) and measure according to the 2020 edition of the Chinese Pharmacopoeia, Part IV, General Rules 2201 (Extract Determination-Cold Immersion Method), about 4g of each batch, accurately weighed, placed in a 250mL conical flask, accurately add 100mL of 95% ethanol, plugged, cold soaked, shaken from time to time within the first 6 hours, then let it stand for 18 hours, quickly filtered with a drying filter, accurately measured 20mL of the filtrate, placed in an evaporating dish that has been dried to constant weight, evaporated to dryness on a water bath, dried at 105°C for 3 hours, cooled in a desiccator for 30 minutes, and quickly and accurately weighed. Unless otherwise specified, the content (%) of alcohol-soluble extracts in the test sample is calculated based on the dry product. The results are shown in Table 3.

[0098] Table 3 Determination results of extracts from three batches of Wuzhimaotao Buxu granules

[0099] batch number Extract (%) 230901 10.25 230902 10.22 230903 10.21

[0100] 1.4 Microbial limits

[0101] Inspect according to the microbial limit test method for non-sterile products (General Rules 1105, 1106, and 1107 of Part IV of the 2020 edition of the Chinese Pharmacopoeia).

[0102] According to the microbial limit test standard of Wuzhimaotao Buxu Granules established above, three batches of samples (230901, 230902, and 230903) were tested, and the results are shown in Table 4.

[0103] Table 4 Results of microbial limit test of Wuzhimaotao Buxu granules

[0104]

[0105] 5. Establishment of extraction process evaluation method

[0106] In this experiment, the dry paste rate was selected as the evaluation index of the extraction process.

[0107] 5.1 Determination of dry paste rate

[0108] Measure the volume of the extracted medicinal solution, then accurately pipette 50 mL into an evaporating dish dried to constant weight. Evaporate to dryness in a water bath, then dry in a 105°C electric constant-temperature drying oven for 5 hours. Transfer to a desiccator, cool for 30 minutes, accurately weigh, and dry again at the same temperature for 1 hour. Cool and weigh again until the difference between two consecutive weighings is no more than 5 mg. Calculate the dry paste yield according to the following formula.

[0109] Dry paste rate (%) = (dry paste mass × total volume of liquid medicine) / (total mass of medicinal materials × sampling volume) × 100%

[0110] 6. Water absorption test

[0111] Dried Chinese medicinal materials will absorb a certain amount of water. In order not to affect the changes in the subsequent amount of water added, the water absorption rate is first measured. Based on this, when adding water for the first time, more water equal to the water absorption rate is added to reduce the error caused by water absorption of the medicinal materials.

[0112] Take the daily prescribed amount of medicinal materials, weigh them, and soak them in 10 times the amount of water. Check every 2 hours whether the rhizome medicinal materials are soaked. After the medicinal materials are soaked, filter them until no more liquid drops. Weigh the mass of the wet medicinal materials and calculate the water absorption rate according to the following formula. The results are shown in Table 5.

[0113] Water absorption rate (%) = (wet medicinal material mass - dry medicinal material mass) / dry medicinal material mass × 100%

[0114] Table 5 Test results of water absorption rate of medicinal materials

[0115]

[0116] From the results, we can see that the water absorption rate of the medicinal materials after soaking is 157.76%, which is about twice the weight of the medicinal materials. Therefore, when adding water for the first time, it is necessary to add twice the amount of water of the medicinal materials.

[0117] 7. Single factor investigation

[0118] Single-factor experiments examine the impact of one experimental factor on evaluation indicators by varying it while keeping the others fixed. Through single-factor experiments, the factors and levels of the orthogonal experiment are determined, leading to the optimization of the extraction process. Common factors influencing the aqueous extraction process of traditional Chinese medicines include soaking time, extraction time, water addition, and the number of decoctions. Decoctions are generally performed 1, 2, or 3 times, so single-factor experiments primarily examine the effects of soaking time, extraction time, and water addition on evaluation indicators.

[0119] 7.1 Immersion time investigation

[0120] Take 6 portions of the daily prescription medicinal materials, add 10 times the amount of water respectively, soak for 0, 0.5, 1.0, 1.5, 2.0, and 2.5 hours, heat and extract for 1.0 hour, filter, measure the volume of the filtrate, and then calculate the dry paste rate under each condition according to the method under "5.1". The results are shown in Table 6.

[0121] Table 6 Soaking time investigation results (n=3)

[0122] Soaking time / h Dry paste rate / % 0 24.68±1.50 0.5 29.37±0.89 1.0 29.82±2.89 1.5 30.51±1.87 2.0 31.09±1.20 2.5 31.85±1.09

[0123] The results show that the increase in soaking time has little effect on the dry paste rate, so soaking time is not considered as a factor in the optimization of the extraction process. However, in actual production, the amount of medicinal materials is large, and in order to allow the medicinal materials to absorb water thoroughly, the soaking time before extraction is determined to be 0.5h.

[0124] 7.2 Investigation of water addition

[0125] Take 6 portions of the daily prescription amount of medicinal materials, add 4, 6, 8, 10, 12, and 14 times the amount of water respectively, soak for 0.5 hours, heat and extract for 1.0 hours, filter, measure the volume of the filtrate, and then calculate the dry paste rate under each condition according to the method under "5.1". The results are shown in Table 7.

[0126] Table 7 Results of water addition (n=3)

[0127] Water addition / times Dry paste rate / % 4 23.78±2.53 6 27.75±1.15 8 29.33±1.43 10 30.85±2.01 12 31.71±1.72 14 32.17±2.10

[0128] The results show that the change in the amount of water added has a great influence on the dry paste rate. When the amount of water added reaches 10 times, the increase in the dry paste rate tends to be gentle. Therefore, the amount of water added is selected as the investigation factor for the optimization of the extraction process, and the levels are selected as 6, 8, and 10 times.

[0129] 7.3 Extraction time investigation

[0130] Take 6 portions of the daily prescription amount of medicinal materials, add 10 times the amount of water to each, soak for 0.5 h, heat and extract for 0.5, 1.0, 1.5, 2.0, 2.5, and 3.0 h, filter, measure the volume of the filtrate, and then calculate the dry paste rate under each condition according to the method under "5.1". The results are shown in Table 8.

[0131] Table 8 Extraction time investigation results (n=3)

[0132] Extraction time / h Dry paste rate / % 0.5 28.77±1.69 1.0 32.06±1.81 1.5 32.93±2.01 2.0 33.02±2.33 2.5 33.42±1.92 3.0 33.35±2.05

[0133] The results show that the increase in extraction time has a greater impact on the dry paste rate. After the extraction time reaches 1.5h, the increase in the dry paste rate tends to be gentle. Therefore, the extraction time is selected as the investigation factor for the optimization of the extraction process, and the levels are selected as 0.5, 1.0, and 1.5h.

[0134] 8. Orthogonal Design

[0135] 8.1 Factor Level Design

[0136] According to the results of single factor investigation and combined with production practice, the amount of water added (A), extraction time (B), and number of extractions (C) were selected as the investigation factors of the orthogonal experiment. The levels of orthogonal design factors are shown in Table 9.

[0137] Table 9 Factor level table

[0138]

[0139] 8.2 Orthogonal experiment

[0140] The dry extract ratio was used as the evaluation index, and the optimal extraction process was determined based on the dry extract ratio evaluation results. The orthogonal test arrangement and results are shown in Table 10, and the variance analysis is shown in Table 11.

[0141] Table 10 Orthogonal test arrangement and results

[0142]

[0143] Table 11 Analysis of variance of dry paste rate

[0144] According to the results of variance analysis, factor C (number of extractions) had a significant effect on the extraction process, and factors A (amount of water added)

[0145] Sources of Error SS f S F P A 17.87 2 8.94 33.97 <0.05 B 17.12 2 8.56 32.51 <0.05 C 280.00 2 140 531.81 <0.01 error 0.53 2 0.26

[0146] Factor B (extraction time) significantly impacted the extraction process, with the order of influencing factors being: C > A > B, meaning number of extractions > amount of water added > extraction time. Based on intuitive analysis, the optimal process combination for this product was identified as: A3B3C3. To conserve energy, adapt to large-scale industrial production, and ensure consistency in the material base by combining traditional decoction methods, this experiment determined the optimal process combination to be: A2B2C2. Specifically, each herbal ingredient was weighed, soaked in water for 30 minutes, and then heated and extracted twice, adding 10 times the amount of water for the first extraction and 8 times the amount for the second extraction, with each extraction lasting 1 hour.

[0147] 8.3 Extraction process validation

[0148] Process validation was conducted based on the orthogonal optimization results. The daily prescribed amount of medicinal materials was weighed and soaked in water for 30 minutes. Heat-extracted twice, adding 10 times the amount of water for the first extraction and 8 times the amount of water for the second extraction. Each extraction lasted 1 hour. Filtered, the filtrates were combined, and the volume of the filtrates was measured. The dry extract yield was calculated according to the method in "5.1." The results are shown in Table 12.

[0149] Table 12 Extraction process verification results

[0150] Test number Dry paste rate / % 1 34.35 2 34.82 3 34.10 average value 34.42

[0151] The results of extraction process verification showed that the average dry paste rate was 34.52% when the extraction was carried out according to the optimized extraction process, which was an excellent result, indicating that the above process was reasonable and feasible.

[0152] 8.4 Conclusion: Through single factor experiment, orthogonal experiment and verification experiment, the extraction process of Wuzhimaotao Buxu Granules was finally determined as follows: weigh each medicinal material in the prescription, soak in water for 30 minutes, heat and extract twice, add 10 times the amount of water for the first time and 8 times the amount of water for the second time, and extract for 1 hour each time.

[0153] 9. Concentration process selection

[0154] Currently, the most commonly used concentration methods are atmospheric pressure concentration and vacuum concentration. Vacuum concentration is time-saving, highly efficient, and has a low boiling point. It can reduce the decomposition of heat-sensitive substances and is conducive to retaining effective ingredients. It is the preferred concentration method for large-scale production of traditional Chinese medicine preparations. Combined with the actual production conditions and preliminary test results of our center, it was determined that the concentration method for this product extract adopts double concentration. The first concentration is vacuum concentration at a concentration temperature of 75-95°C. The filtrate is concentrated into a clear paste with a relative density of 1.05-1.15 (60°C); the second concentration is atmospheric pressure concentration. The clear paste is concentrated into a thick paste with a relative density of 1.28-1.35 (60°C).

[0155] 10. Molding process research

[0156] 10.1 Selection of excipients

[0157] The selection of excipients should be based on the safety, effectiveness, and stability of the preparation. Because Chinese herbal extract powder has strong hygroscopicity, this preparation uses dextrin as a filler due to its large capacity, low viscosity, and stable properties.

[0158] 10.2 Selection of ethanol concentration

[0159] The purpose of adding ethanol during granulation is to act as a binder or lubricant. To produce good granules, an appropriate concentration of ethanol should be added so that the granules can form a ball when held, fall apart when lightly pressed, and remain sticky on the palm. Therefore, the ethanol concentration should be determined based on the granule's formability and qualified rate.

[0160] In this study, 1500 g of each of three fine powders was weighed and added to each of the three portions of ethanol at appropriate concentrations based on granule formability. The mixture was then mixed to prepare a soft material. The soft material was then placed in an oscillating granulator and granulated through a 10-mesh sieve. The resulting granules were then dried in an 85°C oven until the moisture content was less than 4.0%. The results are shown in Table 13.

[0161] Table 13 ethanol concentration screening

[0162] Test No. Ethanol concentration (%) Granulation conditions 1 80 The soft material is relatively wet and easy to stick to the screen, the particle qualification rate is low, and the formability is good. 2 85 Soft material is suitable, easier to granulate, and has good granule formability. 3 95 The soft material is loose, the particles are too fine, and the formability is poor.

[0163] The results show that when the ethanol concentration is 85%, the granulation is good, the molding rate is high, and the fluidity is good.

[0164] 11. Preparation prescription

[0165] 11.1 Calculation of Extract Yield

[0166] From the pilot scale verification results (see Table 14), it can be seen that the average actual dry paste yield of 12 medicinal herbs including this variety of five-fingered ginseng is 26.3%.

[0167] 11.2 Determination of Daily Dosage and Filling Size

[0168] The daily dosage of this product is 76g, and the extract yield is 26.1% to 26.6%. The daily dosage of dry extract is 76×26.3%=19.988g. Adding 1:(1.22-1.27) dextrin, the daily dosage is about 44.97g (19.988×2.25=44.97g). Take it three times a day, and the dosage each time is 44.97 / 3=14.99g.

[0169] 12. Pilot production research

[0170] Take by weighing five finger hair peach deficiency-tonifying prescription medicinal material 45.6kg, add water soak after 30 minutes, heating extraction secondary, add 10 times of water gagings for the first time, second, add 8 times of water gagings, extract 1 hour at every turn, 100 mesh strainer filters, merging filtrate, filtrate decompression concentration, concentration temperature is 75~95 ℃, be condensed into the concentrated solution of relative density 1.05~1.15 (60 ℃), be condensed into the thick paste of relative density 1.28~1.35 (60 ℃) by normal pressure again, thick paste and appropriate dextrin mix in trough mixer, be transferred to 65~85 ℃ of controlled temperature in the baking oven, oven dry, dry soft material is ground into fine powder, add appropriate 85% ethanol and moisten, prepare soft material, soft material places oscillating granulator and crosses 10 mesh sieves granulation, the granule that makes places baking oven and is dried to moisture and is lower than 4.0%, takes out, granulate, promptly gets finished product granule. Finished product granule places granule packing machine and packs by 15g / bag. Three batches were produced continuously according to the above production process. The batch numbers of the obtained granules were 230901, 230902, and 230903, respectively. The results are shown in Table 14.

[0171] Table 14 Results of three batches of pilot production

[0172]

[0173] The pilot results show that the yield of the three batches of Wuzhimaotao Buxu granules is greater than 80%, indicating that the determined extraction process and preparation process are reasonable, stable and suitable for production.

[0174] 13. Conclusion: According to the clinical medication needs, single-factor experiment and orthogonal experiment were used, with the dry paste rate as the evaluation index to determine the extraction and concentration process parameters of Wuzhimaotao Buxu Granules. Then, single-factor experiment was used, with the granule forming rate, moisture absorption rate, fluidity and solubility as the evaluation indexes to screen the granulation process. Finally, three batches of pilot production were carried out to verify the preparation process of Wuzhimaotao Buxu Granules. The results showed that the determined preparation process was scientific, reasonable and feasible.

[0175] 14. Stability test of Wuzhimaotao tonic granules

[0176] 14.1 Accelerated test

[0177] 14.1.1 Sample: Three batches of Wuzhimaotao Buxu Granules (Batch No. 230901)

[0178] 14.1.2 Test equipment: Drug stability test chamber

[0179] 14.1.3 Inspection method: Samples are placed in a drug stability test chamber at a temperature of (40±2)°C and a relative humidity of 75%±5% for 6 months. Samples are taken at the end of 0, 3, and 6 months for testing of properties, identification, filling volume, solubility, particle size, moisture, extractables, and microbial limits.

[0180] 14.1.4 Inspection Period: December 2023 - June 2024

[0181] 14.1.5 Inspection results: See Table 15 for details

[0182] 14.2 Long-term testing

[0183] 14.2.1 Sample: Three batches of Wuzhimaotao Buxu Granules (Batch No. 230901)

[0184] 14.2.2 Test equipment: Drug stability test chamber

[0185] 14.2.3 Inspection method: The samples are placed in a drug stability test chamber at a temperature of (25±2)°C and a relative humidity of 60%±10% for 12 months. Samples are taken at the end of 0, 3, 6, 9, 12, and 18 months, and their properties, identification, filling volume, solubility, particle size, moisture, extractables, and microbial limits are tested.

[0186] 14.2.4 Inspection Period: December 2023-June 2025

[0187] 14.2.5 Inspection results: See Table 16 for details

[0188] 14.3 Test Conclusion

[0189] After 6 months of accelerated testing and 18 months of long-term stability testing, the results showed that all quality inspection indicators of this product met the requirements of the draft internal control agent standard and had good stability.

[0190] Table 15 Wuzhimaotao Buxu Granule (Batch No. 230901) accelerated test results

[0191]

[0192]

[0193] Table 16 Long-term test results of Wuzhimaotao Buxu Granules (Batch No. 230901)

[0194]

[0195]

[0196] 15. Pharmacological Test Report of Wuzhimaotao Buxu Granules

[0197] 15.1 Test samples

[0198] The test drug was a thick paste of Wuzhimaotao Buxu Granules, which is used to prepare Wuzhimaotao Buxu Granules. It has the effects of invigorating qi and strengthening the spleen, regulating the liver and kidneys, and is mainly used to treat sub-health, spleen and stomach qi deficiency, liver and kidney deficiency, and qi sinking syndrome. Each gram of the paste is equivalent to 2.19g of the original medicinal material, provided by Guangxi International Zhuang Medicine Hospital, batch number 20230901. The clinical dosage of Wuzhimaotao Buxu Granules is: oral administration of one bag (15g / bag) three times a day (equivalent to taking 57.015g of the original medicinal material daily). According to the dosage conversion method in the "Methodology of Pharmacological Research in Traditional Chinese Medicine" (3rd edition), a dose for rats can be 3 to 7 times the clinical dose, and 10 to 15 times the clinical dose for mice. In this experiment, 6 times the clinical dose was selected as the median dose for rats, and 10 times the clinical dose was selected as the median dose for mice. The dosages are shown in the table below.

[0199] Table 17 Dosage of Wuzhimaotao Buxu Granule Paste for Rats

[0200]

[0201] Table 18 Dosage of Wuzhimaotao Buxu Granule Paste for Oral Administration in Mice

[0202]

[0203] 15.2 Experimental Materials

[0204] 15.2.1 Animals and Experimental Conditions

[0205] Sprague-Dawley rats (SPF grade) were provided by Hunan Slake Jingda Laboratory Animal Co., Ltd. (license number SCXK(Xiang)2019-0004). Animals were housed in groups in a barrier environment system with automated water and feeding, and fed a complete solid diet. The laboratory temperature was maintained at 20–23°C and the relative humidity at 49–65%.

[0206] KM mice, SPF grade, were provided by the Experimental Animal Center of the Guangxi Zhuang Autonomous Region Institute for Drug Control, production license number SCXK-Gui2022-0001. They were housed in groups in an IVC (barrier environment) with automated water and feeding, and fed a complete solid diet. The laboratory temperature was 20–22°C, and the relative humidity was 49–60%.

[0207] Use license number: SYXK-GUI2023-0005.

[0208] 15.2.2 Drugs and other materials

[0209] Reserpine, Sigma-Aldrich, batch number 1001005091; D-xylose, Aladdin Reagent Co., Ltd., batch number L2209415; p-bromoaniline, Shandong Xiya Chemical Co., Ltd., batch number L2209415; rat gastrin (Gas) kit, Shanghai Jianglai Biotechnology Co., Ltd., batch number 120416005129421204; rat motilin (MTL) kit, Shanghai Jianglai Biotechnology Co., Ltd., batch number 12041 6005123551204; rat somatostatin (SS) kit, Shanghai Jianglai Biotechnology Co., Ltd., batch number 120416005129191204; rat vasoactive intestinal peptide (VIP) kit, Shanghai Jianglai Biotechnology Co., Ltd., batch number 120416005129421204; atropine sulfate injection, Guilin Nanyao Co., Ltd., batch number SA6220602; adenine, Shanghai Yien Chemical Technology Co., Ltd., batch number RH441 977; rat immunoglobulin G (IgG) kit, Wuhan Merck Biotechnology Co., Ltd., batch number kt40064; rat immunoglobulin A (IgA) kit, Wuhan Merck Biotechnology Co., Ltd., batch number kt30383; creatinine (CRE) assay kit, Beijing Leadman Biochemical Co., Ltd., batch number 23072710; uric acid (UA) assay kit, Beijing Leadman Biochemical Co., Ltd., batch number 23060210; urea (UREA) assay kit, Beijing Leadman Biochemical Co., Ltd., batch number 22101403; mouse testosterone (T) kit, Wuhan Merck Biotechnology Co., Ltd., batch number kt21086; mouse cortisol (Cortisol) kit, Wuhan Merck Biotechnology Co., Ltd., batch number kt50097; hematoxylin staining solution, Zhuhai Beso Biotechnology Co., Ltd., batch number C230505; eosin staining solution, Zhuhai Beso Biotechnology Co., Ltd., batch number C230803. The relevant reagents were prepared by the Pharmacology Laboratory of our hospital.

[0210] Positive drug for animal experiment: Jinkuishenqi Pills, manufactured by Beijing Tongrentang Technology Co., Ltd., batch number 1001005091 (the human dose is 0.2 g / kg, the mouse dose is 20 times the human dose, i.e. 4 g / kg; the rat dose is 10 times the human dose, i.e. 2 g / kg); preparation method: grind 10 Jinkuishenqi Pills, and add purified water to 300 mL to obtain a 0.2 g / mL solution.

[0211] Test methods and results

[0212] 15.3 Effects of Reserpine-Induced Spleen Deficiency in Rats

[0213] Pick Sixty Sprague-Dawley rats, weighing 240-260 g, were randomly divided into six groups of 10 rats each: a blank control group, a model control group, a positive control group, a low-dose 5-fingered licorice group, a medium-dose 5-fingered licorice group, and a high-dose 5-fingered licorice group. Jinkuishenqi Pills were used as the positive drug. All groups, except the blank control group, received a subcutaneous injection of 0.5 mg / kg reserpine once daily for 15 consecutive days. After fasting for 12 hours, each rat was gavaged with 4 mL of a 3% D-xylose solution. Urine was then collected for 5 hours (during the fasting period, water was not contraindicated). A 0.5 mL urine sample was collected from each rat, diluted 10-fold with double-distilled water, and 2.5 mL of a 2% p-bromoaniline solution was added. The sample was placed in a 70°C water bath for 10 minutes and then at room temperature for 70 minutes. The absorbance at 520 nm was measured, and the xylose excretion rate was calculated. After successful model establishment, the rats were grouped according to body weight and xylose excretion rate. The results are shown in Table 19. The five-fingered ginseng group was gavaged with different doses of five-fingered ginseng tonic granule solution, the positive control group was gavaged with Jinkui Shenqi pill solution, and the blank control group and spleen deficiency model control group were gavaged with distilled water, once a day for 14 consecutive days, with a gavage volume of 10 mL / kg, and changes in body weight and physical signs were recorded every other day. Before the end of the experiment, the rats were fasted but not watered for 12 hours, and the xylose excretion rate was detected according to the above method. At the end of the experiment, blood was collected from the abdominal aorta, and serum was separated. The enzyme-linked immunosorbent assay (ELISA) was used to determine the levels of gastrin (Gas), motilin (MTL), somatostatin (SS), and vasoactive intestinal peptide (VIP) in the serum. The data were statistically analyzed using the one-way analysis of variance-LSD method in IBM SPSS Statistics25.0 software. The results are shown in Table 20. Figure 1-Figure 3 .

[0214] Table 19 Effects of reserpine on body weight and xylose excretion rate in rats

[0215] Group n Weight (g) Xylose excretion rate (%) Blank control group 10 354.24±9.61 32.00±10.90 Model control group 10 280.94±27.17** 16.30±11.33** Positive control group 10 283.17±24.99** 16.61±13.15** Low-dose group 10 284.38±23.95** 16.48±11.07** Medium dose group 10 283.80±25.39** 16.81±12.47** High-dose group 10 283.30±22.77** 16.10±12.47**

[0216] Note: Compared with the blank control group, **P<0.01, the rest P>0.05

[0217] The model rats showed mushy loose stools, fatigue, laziness, decreased appetite, and their body weight and xylose excretion rate were significantly lower than those in the blank control group (p<0.01), which is consistent with the characteristics of spleen qi deficiency syndrome, indicating that the model was successful.

[0218] Table 20 Effects on xylose excretion rate and gastrointestinal hormone secretion in rats with spleen deficiency induced by reserpine ( n=10)

[0219]

[0220] Note: Compared with the blank control group, #P<0.05, ##P<0.01; compared with the model control group, *P<0.05, **P<0.01, and the rest are P>0.05

[0221] Xylose absorption is a key indicator of spleen deficiency, and urinary excretion reflects the relative absorption of the small intestine. GAS promotes the secretion of gastric acid and pepsin, stimulates gastrointestinal motility, increases mucosal blood flow, and nourishes the gastrointestinal mucosa. MTL activates gastrointestinal smooth muscle, promotes digestive tract motility, accelerates gastric emptying, and stimulates the secretion of pepsin and pancreatic juice. SS inhibits gastric acid secretion and gastric motility and relaxes vascular smooth muscle. VIP inhibits the secretion of various gastrointestinal hormones.

[0222] After modeling, the body weights of rats in the positive control group and the Wuzhimaotao group were significantly lower than those in the blank control group. Compared with the model control group, there were no significant changes in body weight in the positive control group and the Wuzhimaotao group at all doses (p>0.05). Compared with the blank control group, the model control group showed significantly decreased xylose excretion rate, somatostatin (SS), and vasoactive intestinal peptide (VIP) levels (p<0.01), while significantly increased motilin (MTL) and gastrin (Gas) levels (p<0.05). These results suggest decreased small intestinal absorption function and gastrointestinal hormone imbalance in the model control group. After administration of Wuzhimaotao Buxu Granule Qing Gao, compared with the model control group, the high-dose group showed significantly increased xylose excretion rate, somatostatin (SS), and vasoactive intestinal peptide (VIP) levels (p<0.05), while motilin (MTL) levels were significantly decreased in the medium- and high-dose groups (p<0.05), demonstrating a dose-related relationship. Although gastrin levels after administration were not statistically significant compared with the model control group, they did show a downward trend. The results showed that Wuzhimaotao Buxu Granules could improve the gastrointestinal absorption function and regulate the gastrointestinal hormone levels in the spleen deficiency model control group rats.

[0223] 15.4 Effects on Gastric Emptying and Small Intestinal Propulsion in Mice

[0224] 15.4.1 Effects on Gastric Emptying and Small Intestinal Propulsion in Normal Mice

[0225] 50 mice weighing 18-22 g were taken. The mice were randomly divided into 5 groups, each with 10 mice, including a blank control group, a positive control group, a low-dose group of Wuzhimaotao, a medium-dose group of Wuzhimaotao, and a high-dose group of Wuzhimaotao. Jinkuishenqi Pills were used as the positive drug. The Wuzhimaotao group was given different doses of Wuzhimaotao Buxu Granule solution by gavage, the positive control group was given Jinkuishenqi Pill solution by gavage, and the blank control group was given distilled water by gavage for 7 consecutive days, once a day, with a gavage volume of 20 mL / kg. One hour after the last administration, mice that had been fasting (without water deprivation) for 18 hours were given a semi-solid black paste (10% activated carbon-sodium carboxymethylcellulose solution) by gavage, 0.8 mL per mouse. 20 minutes later, the mice were killed by cervical dislocation, the abdomen was opened, the gastric cardia and pylorus were ligated, the stomach was removed, and the total weight was weighed after wiping it dry with filter paper. The stomach was then cut open along the greater curvature of the stomach, the stomach contents were washed out, and the stomach was wiped dry and weighed. The difference between the total weight of the stomach and the net weight of the stomach was taken as the weight of the residual gastric content, and the percentage of the residual gastric content to the weight of the semisolid paste administered was calculated as the residual gastric rate. Simultaneously, the small intestine was quickly removed, and the mesentery was gently peeled off and the small intestine was pulled into a straight line. The total length of the small intestine from the pylorus to the ileocecal region and the distance from the pylorus to the front edge of the black semisolid paste were measured. The percentage of the distance from the pylorus to the front edge of the black semisolid paste to the total length from the pylorus to the ileocecal region was taken as the small intestine propulsion rate. The data were statistically analyzed using the one-way analysis of variance-LSD method in IBM SPSS Statistics 25.0 software. The results are shown in Tables 21 and Figure 4 .

[0226] Table 21 Effects on gastric emptying and small intestinal propulsion in normal mice ( n=10)

[0227] Group Gastric residual rate (%) Small intestinal propulsion rate (%) Blank control group 23.12±5.91 48.48±6.84 Positive control group 28.72±6.21* 55.54±7.36* Low-dose group 20.24±3.75 46.48±5.49 Medium dose group 29.24±5.21* 50.98±8.10 High-dose group 31.25±7.87** 51.29±9.58

[0228] Note: Compared with the blank control group, *P<0.05, **P<0.01, and the rest P>0.05

[0229] Compared with the blank control group, the gastric residual rate of the medium-dose and high-dose groups of Prunus mume was significantly increased (p<0.05), and had little effect on the small intestinal propulsion rate.

[0230] Effects of 15.5 on gastric emptying and small intestinal propulsive motility in mice with atropine-induced gastrointestinal dysmotility

[0231] 60 mice weighing 18-22 g were taken. The mice were randomly divided into 6 groups, each with 10 mice, including blank control group, model control group, positive control group, low-dose Wuzhimaotao group, medium-dose Wuzhimaotao group and high-dose Wuzhimaotao group. Jinkuishenqi Pills were used as positive drugs. The Wuzhimaotao group was given different doses of Wuzhimaotao Buxu Granules solution by gavage, the positive control group was given Jinkuishenqi Pills solution by gavage, and the blank control group was given distilled water by gavage for 7 consecutive days, once a day, with a gavage volume of 20 mL / kg. 0.5 hours after the last administration, 2.5 mg / kg of atropine sulfate injection was injected intraperitoneally. 0.5 hours later, the mice that had been fasting (without water) for 18 hours were given semi-solid black paste (10% activated carbon-sodium carboxymethyl cellulose solution) by gavage, 0.8 mL per mouse. After 20 minutes, the mice were killed by dislocating the cervical vertebra, the abdomen was opened, the gastric cardia and pylorus were ligated, the stomach was removed, and the total weight was weighed after wiping it dry with filter paper. Then, the stomach body was cut open along the greater curvature of the stomach, the stomach contents were washed out and wiped dry, and the net weight was weighed. The difference between the total weight of the stomach and the net weight of the stomach was taken as the weight of the residual gastric content, and the percentage of the residual gastric content to the weight of the semi-solid paste injected was calculated as the residual gastric rate. At the same time, the small intestine was quickly removed, and the mesentery was gently peeled off and the small intestine was pulled into a straight line. The total length of the small intestine from the pylorus to the ileocecal region and the distance from the pylorus to the front of the black semi-solid paste were measured. The percentage of the distance from the pylorus to the front of the black semi-solid paste to the total length from the pylorus to the ileocecal region was taken as the small intestine propulsion rate. The data were statistically analyzed using the one-way analysis of variance-LSD method in IBM SPSS Statistics 25.0 software. The results are shown in Tables 22 and Figure 5 .

[0232] Table 22 Effects on gastric emptying and small intestinal propulsion in mice with gastrointestinal motility disorders ( n=10)

[0233] Group Gastric residual rate (%) Small intestinal propulsion rate (%) Blank control group 35.44±9.28 50.15±12.33 Model control group 22.20±5.20# 38.23±13.65# Positive control group 38.15±16.77** 52.44±14.62* Low-dose group 21.85±8.89 45.02±11.18 Medium dose group 31.24±13.16 49.23±12.45 High-dose group 41.46±10.83** 54.07±12.94**

[0234] Note: Compared with the blank control group, #P<0.05; compared with the model control group, *P<0.05, **P<0.01, and the rest are P>0.05

[0235] Atropine sulfate, an M receptor blocker, can inhibit normal gastrointestinal function and cause gastrointestinal motility disorders. Compared with the blank control group, the gastric residual rate and small intestinal propulsion rate in the model control group were significantly reduced (p < 0.05). Compared with the model control group, the high-dose Wuzhimaotao group significantly increased the gastric residual rate and small intestinal propulsion rate (p < 0.01). This suggests that Wuzhimaotao Buxu Granules Paste can counteract atropine-induced gastrointestinal motility disorders.

[0236] 15.6 Effect of Adenine on Kidney Yang Deficiency in Rats

[0237] Pick Sixty SD rats weighing 200±20g were fed adaptively for 6 days. They were divided into a blank control group (10 rats) and a modeling group (50 rats) according to body weight balance and random number table method. Except for the blank control group, the other groups were gavaged with 150mg / (kg·d) sodium adenine carboxymethylcellulose (CMC-Na) solution for 21 consecutive days to establish a rat model of kidney yang deficiency nephropathy. According to the theory of traditional Chinese medicine, the rats were observed to see if they had the following symptoms: (1) mental depression and fear of cold; (2) reduced activity and curling up; (3) loose stools, clear urine, and dirty perianal area; (4) loose and dull body hair. The kidney yang deficiency symptoms of the rats in each group were scored according to the above symptoms: no symptoms were scored as 0, 1 symptom was scored as 1, 2 symptoms were scored as 2, 3 symptoms were scored as 3, and 4 symptoms were scored as 4. The model was considered successful if the kidney yang deficiency symptom score of the rats was greater than 0. After modeling, the rats were randomly divided into a model control group, a positive control group, and a low-, medium-, and high-dose Wuzhimaotao group according to body weight balance using a random number table, with 10 rats in each group. Jinkuishenqi Pills were used as the positive drug. The low-, medium-, and high-dose Wuzhimaotao groups, as well as the positive control group, were gavaged with 2.85 g, 5.7 g, and 11.4 g of Wuzhimaotao Buxu Granules, respectively, and 2 g / kg of Jinkuishenqi Pills. The model control group and the blank control group were gavaged with 10 mL / kg of 1% CMC-Na solution for 14 consecutive days. Rats in each group were weighed weekly and their coat color was observed. Symptoms of kidney yang deficiency were scored one day before the end of treatment. After administration, the subjects were anesthetized with 0.4% sodium pentobarbital, and blood was collected from the abdominal aorta. Serum IgA and IgG levels were determined by enzyme-linked immunosorbent assay (ELISA). Serum creatinine (Cr), urea nitrogen (UREA), and uric acid (UA) levels were measured using an automatic biochemical analyzer. Bilateral kidneys were weighed, and organ coefficients were calculated. Changes in renal tissue were observed using HE staining. Data were statistically analyzed using one-way analysis of variance (ANOVA)-LSD method using IBM SPSS Statistics 25.0 software. The results are shown in Tables 23-25. Figure 6-Figure 7 .

[0238] Table 23 Effects of adenine on rat body weight and symptom scores ( n=10)

[0239]

[0240]

[0241] Note: Compared with the blank control group, **P<0.01, the rest P>0.05

[0242] All modeled rats showed weight loss and symptoms of kidney yang deficiency, and the body weight and kidney yang deficiency symptom scores were significantly lower than those of the blank control group (p<0.01), which was consistent with the characteristics of kidney yang deficiency syndrome, indicating that the model was successful.

[0243] Table 24 Effects of adenine on body weight, symptom score and renal organ coefficient in rats with kidney yang deficiency ( n=10)

[0244]

[0245] Note: Compared with the blank control group, #P<0.05, ##P<0.01; compared with the model control group, *P<0.05, **P<0.01, and the rest are P>0.05

[0246] Table 25 Effects of adenine on serum parameters of rats with kidney yang deficiency ( n=10)

[0247]

[0248]

[0249] Note: Compared with the blank control group, #P<0.05, ##P<0.01; compared with the model control group, *P<0.05, **P<0.01, and the rest are P>0.05

[0250] UREA and Cr are key indicators for evaluating the progression and staging of chronic kidney disease and are also important indicators for evaluating the effectiveness of treatment for chronic renal failure. UA levels in the blood are a key indicator of renal function. Immunoglobulins are active serum components produced by plasma cells in response to antigenic stimulation that specifically bind to antigens and are important indicators of human immune function.

[0251] Compared with the blank control group, the kidney yang deficiency symptom score, renal organ coefficient, Cr, UREA, and UA in the model control group were significantly increased (p < 0.01), IgA was significantly decreased (p < 0.05), and IgG showed a downward trend (p < 0.05). These results indicate that the model control group had decreased renal function and immune dysfunction. After administration of Wuzhimaotao Buxu Granule Clearing Paste, compared with the model control group, the kidney yang deficiency symptom score in the high-dose Wuzhimaotao group was significantly reduced (p < 0.01), Cr, UA, and UREA were significantly decreased in the medium-dose and high-dose groups (p < 0.01), and Cr and UA were also significantly decreased in the low-dose group (p < 0.01). IgA was significantly increased in the high-dose group (p < 0.05), and IgG was also significantly increased in the low-, medium-, and high-dose groups (p < 0.05), showing a dose-related effect. This suggests that Wuzhimaotao Buxu Granule Clearing Paste can improve renal function and regulate immune function in the kidney yang deficiency model control group.

[0252] HE staining results showed that the kidney structure of the rats in the blank control group was intact, with clear cortical and medullary structures, and clear glomerular and renal tubular structures. There was no inflammatory cell infiltration or urate crystals. Compared with the blank control group, the kidney volume of the rats in the model control group was enlarged, the renal tubular epithelial cells were atrophied, some renal tubules were dilated, and transparent casts appeared in the tubular lumen; the renal interstitium and renal tubular lumen were accompanied by red blood cells and a large number of inflammatory cells infiltration; renal interstitial fibrous tissue proliferation. After administration of Wuzhimaotao Buxu Granule Qing Gao, no renal tubular dilation was observed, the infiltration of red blood cells and inflammatory cells in the renal interstitium was reduced, the proliferation of renal interstitial fibrous tissue was improved, and some renal tubules were regenerated. Results are shown in Figure 8 .

[0253] 15.7 Effect of Adenine on Kidney Yang Deficiency in Mice

[0254] Pick Sixty Kunming mice weighing 18-22 g were fed adaptively for 6 days. They were divided into a blank control group (10 mice) and a modeling group (50 mice) according to body weight balance and random number table method. Except for the blank control group, the other groups were gavaged with 50 mg / (kg·d) sodium adenine carboxymethylcellulose (CMC-Na) solution for 21 consecutive days with a gavage volume of 20 mL / kg to establish a mouse model of kidney yang deficiency and nephropathy. According to the theory of traditional Chinese medicine, the mice were observed to see if they showed the following symptoms: (1) mental depression and fear of cold; (2) reduced activity and curling up; (3) loose stools, clear urine, and dirty perianal area; (4) loose and dull body hair. The symptoms of kidney yang deficiency in each group of mice were scored according to the above symptoms: no symptoms were scored as 0, 1 symptom was scored as 1, 2 symptoms were scored as 2, 3 symptoms were scored as 3, and 4 symptoms were scored as 4. A score of kidney yang deficiency symptom greater than 0 was considered a successful modeling. After the modeling was completed, the modeling group mice were randomly divided into model control group, positive control group, and five-fingered ginseng low-, medium-, and high-dose groups according to body weight balance and random number table method, with 10 mice in each group, and Jinkuishenqi Pills were used as the positive drug. The five-fingered ginseng low-, medium-, and high-dose groups and positive control group were gavaged with 4.75g crude drug / kg, 9.5g crude drug / kg, and 19g crude drug / kg of five-fingered ginseng tonic granules, and 2g / kg of Jinkuishenqi Pills, respectively. The model control group and the blank control group were gavaged with 20mL / kg of 1% CMC-Na solution for 14 consecutive days. During this period, the body weight of each group of mice was weighed every week, and the changes in coat color were observed. One day before the end of the administration, the symptoms of kidney yang deficiency in each group of mice were scored. After the end of the administration, the mice were anesthetized with 0.4% sodium pentobarbital, the eyeballs were removed to collect blood, and the levels of testosterone (T) and cortisol (cortisol) in the serum were determined by enzyme-linked immunosorbent assay (ELISA). The results are shown in Tables 26-28 and Figure 9 .

[0255] Table 26 Effects of adenine on body weight and symptom scores of mice ( n=10)

[0256]

[0257]

[0258] Note: Compared with the blank control group, **P<0.01, the rest P>0.05

[0259] All modeled mice showed weight loss and symptoms of kidney yang deficiency, and their weight and kidney yang deficiency symptom scores were significantly lower than those of the blank control group (p<0.01), which was consistent with the characteristics of kidney yang deficiency syndrome, indicating that the model was successful.

[0260] Table 27 Effects of adenine on body weight and symptom scores of mice with kidney yang deficiency ( n=10)

[0261] Group Body weight at the end of the experiment (g) Kidney Yang deficiency symptom score at the end of the trial (points) Blank control group 37.45±2.97 0 Model control group 30.03±3.83## 2.5±0.85# Positive control group 32.07±3.53 1.9±0.32* Low-dose group 32.75±4.12 2.2±0.63 Medium dose group 30.50±3.18 2.0±0.67* High-dose group 31.21±3.18 1.8±0.42**

[0262] Note: Compared with the blank control group, #P<0.05, ##P<0.01; compared with the model control group, *P<0.05, **P<0.01, and the rest are P>0.05

[0263] Table 28 Effects of adenine on serum parameters of mice with kidney yang deficiency ( n=10)

[0264] Group T (nmol / L) Cortisol (μg / L) Blank control group 1.50±0.20 32.91±7.52 Model control group 1.24±0.10# 43.82±17.43# Positive control group 1.50±0.10* 30.95±13.10* Low-dose group 1.34±0.18 35.64±9.86 Medium dose group 1.38±0.39 33.40±9.64 High-dose group 1.59±0.46** 27.05±10.22**

[0265] Note: Compared with the blank control group, #P<0.05, ##P<0.01; compared with the model control group, *P<0.05, **P<0.01, and the rest are P>0.05

[0266] Kidney Yang Qi can be reflected by sex hormone levels, with T levels significantly reduced in patients with Kidney Yang Deficiency. Cortisol is a steroidal glucocorticoid, whose secretion is controlled by adrenocorticotropic hormone (ACH) from the anterior pituitary gland. Serum cortisol levels directly reflect the secretory function of the adrenal cortex. Chronic high levels of cortisol can suppress the immune system, making the body more susceptible to infection and disease. Lowering cortisol can enhance immunity and improve the body's resistance. Excessive cortisol levels can interfere with the digestive system's stagnation function, causing gastrointestinal problems such as acid reflux and indigestion. Lowering cortisol can promote digestion and relieve gastrointestinal discomfort.

[0267] Compared with the blank control group, T levels in the model control group were significantly lower (p < 0.05) and cortisol levels were significantly higher (p < 0.05). These results suggest that the model control group exhibited dysregulation of sex hormone secretion and adrenal cortical secretory function. Following administration of the Wuzhimaotao Buxu Granule Clearing Paste, T levels in the high-dose Wuzhimaotao group were significantly higher, while cortisol levels were significantly lower (p < 0.01), demonstrating a dose-dependent relationship. This suggests that Wuzhimaotao Buxu Granule Clearing Paste can improve hormone levels and adrenal cortical secretory function in mice with kidney yang deficiency.

[0268] Conclusion: The above experimental results show that, when administered orally to the test samples, Wuzhimaotao Buxu Granules can significantly increase the xylose excretion rate, SS and VIP levels in rats with spleen deficiency induced by reserpine, and significantly reduce the MTL level, indicating that Wuzhimaotao Buxu Granules can improve the gastrointestinal absorption function and regulate the gastrointestinal hormone levels in rats in the spleen deficiency model control group; it can increase the gastric residual rate in blank mice and counteract atropine-induced gastrointestinal motility disorders; it can reduce the Cr, UA and UREA levels in rats with kidney yang deficiency induced by adenine, and increase the IgA and IgG levels, indicating that Wuzhimaotao Buxu Granules can improve the renal function and regulate the immune function in rats with kidney yang deficiency model; it can increase the testosterone level and reduce the cortisol level in mice with kidney yang deficiency induced by adenine, indicating that Wuzhimaotao Buxu Granules can improve the hormone levels and adrenal cortex secretion function in mice with kidney yang deficiency.

[0269] The above test results indicate that Wuzhimaotao Buxu Granules have the effects of invigorating Qi and strengthening the spleen, regulating kidney deficiency, regulating gastrointestinal function and immune function, and improving sub-health status, providing a certain animal test basis for the clinical application of this product.

[0270] 16. Acute toxicity test of Wuzhimaotao Buxu granules

[0271] 16.1 Preparation of the test solution: Wuzhimaotao Buxu Granule Paste (provided by Guangxi International Zhuang Medicine Hospital), batch number: 230901. Take 100 g of Wuzhimaotao Buxu Granule Paste, add an appropriate amount of purified water, mix well, and prepare a final volume of 92 mL of the test solution (each 1 mL contains 2.38 g of the original medicinal material).

[0272] 16.2 Test Method: Take 20 mice (♀ After 6 hours of feeding cessation, the mice were gavaged with the test solution at a volume of 30 mL / kg for a total of four doses within 24 hours. Systemic reactions and mortality of the mice were observed during the administration period and up to 14 days after the administration.

[0273] 16.3 Test Results: Within 14 days after administration, there were no obvious abnormalities in the appearance, activity, behavior, eating, excretion, etc. of the mice. There were no deaths, and the weight gain of the mice was normal (see the table below).

[0274] Table 29 Body weight gain in acute toxicity test of Wuzhimaotao Buxu granules in mice

[0275]

[0276] 16.4 Conclusion: The experiment showed that when Wuzhimaotao Buxu Granules were administered to mice orally four times within 24 hours, the maximum dosage was 285.6 g / kg of the original medicinal material (equivalent to 301 times the clinical daily dose) based on body weight. The maximum dosage was 285.6 g / kg of the original medicinal material (equivalent to 301 times the clinical daily dose) based on body surface area (the body surface area of ​​a mouse weighing 20.2 g is 0.00675 m 2 The surface area of ​​a 60kg human body is 1.625m 2 ), the maximum dosage is 854.7g / m 2 (equivalent to 24 times the clinical daily dose); no obvious acute toxicity was observed. This suggests that Wuzhimaotao Buxu Granules may be safe when used according to its clinical usage and dosage.

[0277] 17. Repeated-dose toxicity (long-term toxicity) test report of Wuzhimaotao Buxu granules

[0278] Experimental Methods: Rats were acclimated for 5 days before the experiment and randomly divided according to body weight into a control group and high-, medium-, and low-dose Wuzhimaotao Buxu Granule groups, with 30 rats in each group (half in the ♀ and ♂ groups, 20 in the main experiment and 10 in the recovery period). The high-, medium-, and low-dose groups were administered an aqueous solution of the test drug at doses of 57, 28.5, and 14.3 g / kg of the test drug raw material, respectively, while the control group received purified water. Administration was performed orally once daily at the same time in the morning and afternoon for 30 consecutive days, with a dose volume of 20 mL / kg per dose for each group. During the dosing period, the rats were observed daily for general health and toxic reactions. Body weights were measured weekly, and dosing was adjusted based on body weight. Food intake was measured weekly in each cage. Urine was collected from 20 and 10 rats in each group in metabolic cages at 6:00 PM 3 hours after the last dose and 30 days after the last dose, respectively, for analysis. On the day following the last dose and 31 days after drug withdrawal, 20 and 10 rats from each group (fasted for 15-16 hours) were anesthetized with an intraperitoneal injection of 40 mg / kg sodium pentobarbital. Blood was collected via the abdominal aorta for hematological and biochemical examinations. Some organs were dissected and weighed, and organ coefficients were calculated based on organ weight per 100 g body weight. Major organs were dissected for gross and histopathological examinations. The values ​​in each treatment group were compared with those in the control group, and intergroup t-tests were performed using SPSS 25.0 statistical analysis software.

[0279] Test results: Rats were continuously gavaged with an aqueous solution of Wuzhimaotao Buxu Granule Clear Paste at doses of 57, 28.5, and 14.3 g / kg of the original medicinal material (60, 30, and 15 times the intended clinical daily dose, respectively) for 30 days, and some rats were observed for 30 days after drug withdrawal. During the drug administration period and the drug withdrawal observation period, no obvious abnormal changes were observed in the general condition, eyes, routine hematological examination indicators, urine examination indicators, and histopathological examination of major organs of the rats in the drug administration group. During the drug administration period, the weight gain of the rats was slow and the food intake decreased, which gradually returned to normal after drug withdrawal, and was considered to have no obvious toxicological significance. AST and CK decreased in the drug administration group after administration, and ALP increased in the high-dose group, but these changes were reversible and were considered to have no obvious toxicological significance; no changes were observed in other blood biochemical indicators. After administration, the liver organ coefficients of the high- and medium-dose groups and the kidney organ coefficient of the high-dose group increased, which may be the result of the dual effects of the test drug and the weight loss of rats. However, this effect is reversible and did not cause changes in liver and kidney biochemical indicators or obvious liver and kidney tissue pathological lesions; no gross morphological changes in other organs were observed.

[0280] No significant repeated-dose toxicity or delayed toxicity was observed with the test drug in this trial, suggesting that Wuzhimaotao Buxu Granules may be safe for clinical use according to the prescribed dosage, route, and course of treatment. However, changes in some observed indicators suggest that clinical use of the test drug should pay attention to the patient's liver, kidney, and thyroid function, and medication should be discontinued if significant abnormalities occur.

[0281] The disclosed embodiments are described to enable one skilled in the art to implement or use the present invention. Various modifications to these embodiments will be readily apparent to one skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention is not limited to the embodiments shown herein but is to be construed in the widest possible manner consistent with the principles and novel features disclosed herein.

Claims

1. A five-fingered peach tonic granule, characterized in that: The method is prepared by comprising the following raw materials in parts by weight: 12-20 parts of five-fingered peach, 7-15 parts of codonopsis pilosula, 5-10 parts of atractylodes macrocephala, 5-10 parts of angelica, 4-10 parts of dried tangerine peel, 3-8 parts of bupleurum, 3-8 parts of cimicifuga, 5-10 parts of wolfberry, 5-10 parts of dodder seed, 5-10 parts of psoralea corylifolia, 5-10 parts of epimedium, and 4-10 parts of roasted liquorice.

2. A method for preparing Wuzhimaotao nourishing granules, characterized in that: The following steps are involved: (1) Weigh the raw materials according to the Wuzhimaotao tonic granules according to claim 1; (2) soaking the weighed raw materials in water, decocting them twice to obtain a medicinal solution, and filtering the medicinal solution to obtain a filtrate; (3) concentrating the obtained filtrate for the first time to obtain a concentrated solution, and concentrating the concentrated solution again to obtain a thick paste of Wuzhimaotao Buxu Granules; (4) adding the obtained Wuzhimaotao Buxu Granules paste to the dextrin while stirring, continuing to stir evenly, drying, and crushing to obtain a fine powder; (5) The obtained fine powder is stirred evenly, and 85% ethanol is added to moisten and stir to prepare a soft material, and the soft material is sieved and granulated to obtain wet granules; (6) drying the obtained wet granules, re-granulating the granules, and stirring the granules to obtain the five-fingered ginseng nourishing granules.

3. The method for preparing the Wuzhimaotao nourishing granules according to claim 2, wherein: In step (2), the raw material is soaked in water for 30 minutes and then decocted twice, with 10 times the amount of water added for the first time and 8 times the amount of water added for the second time. Each decoction is for 1 hour. During the decoction process, the steam pressure is controlled at 0.1-0.25 MPa before the medicinal liquid boils, and the steam pressure is controlled at 0.05-0.10 MPa after the medicinal liquid boils, so that the medicinal liquid keeps boiling. After the extraction is completed, the medicinal liquid is filtered with a 100-mesh filter.

4. The method for preparing the Wuzhimaotao nourishing granules according to claim 2, wherein: In step (3), the first concentration temperature is 75-95° C., the vacuum degree is -0.01 to -0.09 MPa, and the concentrate is concentrated to a concentrated liquid with a relative density of 1.05-1.15 at 60° C.; and the concentrate is concentrated again to a thick paste with a relative density of 1.28-1.35 at 60° C.

5. The method for preparing the Wuzhimaotao nourishing granules according to claim 2, wherein: In step (4), the stirring speed is 24 rpm, the stirring time is continued for 5-10 min, the mass ratio of the five-fingered ginseng tonic granule paste and dextrin is 1:(0.8-1.0), the drying temperature is 65-85° C., the mixture is dried to a moisture content of less than 9.0%, and then crushed to a fine powder that can pass through an 80-mesh sieve.

6. The method for preparing the Wuzhimaotao nourishing granules according to claim 2, characterized in that: In step (5), the obtained fine powder is stirred evenly at a stirring speed of 24 rpm for 5-10 min, 85% ethanol is added to moisten and stir to make a soft material until it can be grasped into a ball and falls apart when pressed lightly, and then granulated through a 10-mesh sieve.

7. The method for preparing the Wuzhimaotao nourishing granules according to claim 2, characterized in that: In step (6), the drying temperature is 65-85° C., and the mixture is dried until the moisture content is less than 4.0%. The mixture is sieved with 14-mesh and 60-mesh sieves, and particles that pass through 14-mesh but not 60-mesh are considered qualified particles. The stirring speed is 24 rpm, and the stirring time is 20 minutes.

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