Traditional Chinese medicine composition for treating hypoovarian reserve function and application thereof

Abstract

The invention discloses a traditional Chinese medicine composition for treating hypoovarian reserve function. The traditional Chinese medicine composition comprises the following raw material medicines: 10-30 parts of vinegar tortoise shell, 10-15 parts of prepared rehmannia root, 6-20 parts of salt-processed semen cuscutae, 9-15 parts of wine-processed rhizoma polygonati, 10-30 parts of radix puerariae, 9-15 parts of polygonum cuspidatum, 6-20 parts of acanthopanax, 5-15 parts of salvia miltiorrhiza, 6-10 parts of roasted herba epimedii and 3-10 parts of pericarpium citri reticulatae. The invention further discloses application of the traditional Chinese medicine composition in preparation of a medicine for treating ovarian reserve hypofunction. The traditional Chinese medicine composition has the effects of improving the ovarian function and the sex hormone level, increasing the AFC quantity, adjusting the ovarian blood flow state, improving the menstrual condition, the traditional Chinese medicine syndrome and the anxiety level of a patient and improving the clinical pregnancy rate, and is safe and reliable.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicine, and specifically, to a traditional Chinese medicine composition for treating diminished ovarian reserve and its uses. Background Art

[0002] Diminished ovarian reserve (DOR) refers to the decline in a woman's ovarian ability to produce eggs, the quantity and quality of oocytes, accompanied by disordered sex hormone secretion levels. Menopausal-like symptoms and low pregnancy rates are the main clinical manifestations of DOR. In the long term, it can also increase the risk of diseases such as cardiovascular diseases and osteoporosis, seriously affecting the physical and mental health of patients, family happiness and social stability. The prevalence of DOR is about 10% - 35%, and the incidence rate of women under 40 years old is as high as 10%. Infertility caused by DOR accounts for about 5% of the ovulatory disorder infertility population, and the incidence rate is increasing year by year and showing a younger trend.

[0003] Currently, hormone replacement is the most widely used clinical treatment method for DOR. The drug intervention measures for DOR mainly include: the relatively recognized estrogen-progesterone replacement therapy, such as Femoston, Climen, estradiol valerate + dydrogesterone, etc.; dehydroepiandrosterone (DHEA) that improves antioxidant capacity and coenzyme Q10 that improves mitochondrial function, etc., for which the evidence-based evidence is not yet sufficient. Hormone replacement therapy can improve menstrual disorders and symptoms of sex hormone deficiency, but the effect on improving fertility and regulating the anxiety state is not obvious. When using it, relevant contraindications and precautions should be noted, and its application in clinical practice is subject to certain limitations. For DOR-related infertility, assisted reproductive technologies are often used. Drug-induced ovulation is the main treatment method for ovulatory disorder infertility caused by DOR. However, the incidence rate of ovarian hyporesponse is relatively high, and ovarian function decline affects ovulation, luteal support after pregnancy, endometrial receptivity, etc. Therefore, the treatment has limitations. Treatments such as autologous gametes or in vitro fertilization-embryo transfer with donor eggs, mesenchymal stem cells, and platelet-rich plasma bring problems such as adverse reactions, ethics, law, and the health of offspring, causing many troubles to patients and their families, and have not fundamentally solved the problem of the decline in the quality and / or quantity of oocytes. Although the above are common methods for clinically treating DOR and its related infertility, their clinical application is restricted to a certain extent due to various adverse reactions and contraindications. Patients urgently need a safer and more effective treatment method to improve ovarian reserve function and increase fertility. Traditional Chinese medicine has unique advantages in treating DOR due to its multi-component, multi-target, high safety, etc. characteristics, but there is currently no proprietary Chinese medicine with the indication of treating DOR in China.

[0004] Therefore, for patients with diminished ovarian reserve, there is a need to develop a traditional Chinese medicine preparation that can effectively improve ovarian function, increase fertility, regulate the anxiety state of patients, and improve the overall quality of life of patients. Summary of the Invention

[0005] Professor Cai Lianxiang, a famous traditional Chinese medicine teacher in the capital, believes that kidney essence is the basis of reproduction based on the theory that "kidneys store essence and control reproduction". Kidneys store essence, and essence produces blood. Eggs belong to the reproductive essence in the kidneys. The normal development of follicles depends on the nourishment of essence and blood. If there is insufficient essence and blood in the kidneys, the follicles lack the source of development, and the reproductive cells are gradually consumed, which will lead to DOR. If kidney deficiency is not supplemented for a long time, the degree of ovarian function decline will gradually increase, and the decline in fertility will become increasingly severe. Therefore, Professor Cai emphasizes that early intervention for DOR patients to delay the progression of the disease is necessary to replenish the kidney and essence, and regulate qi and blood to improve the ovarian reserve function of DOR patients, improve the fertility of DOR infertile women, and regulate the anxiety of patients.

[0006] In view of the above technical problems, one object of the present invention is to provide an improved Chinese medicine composition, which can effectively treat diminished ovarian reserve function.

[0007] Another object of the present invention is to provide a pharmaceutical preparation comprising the above-mentioned traditional Chinese medicine composition.

[0008] Another object of the present invention is to provide use of the above-mentioned Chinese medicine composition in the preparation of a medicament for treating ovarian reserve dysfunction and related diseases.

[0009] Another object of the present invention is to provide a method for preparing the above-mentioned Chinese medicine composition.

[0010] In order to achieve the above object, the present invention adopts the following technical solutions:

[0011] In a first aspect, the present invention provides a traditional Chinese medicine composition for treating ovarian reserve dysfunction, which comprises the following raw materials in parts by weight: 10-30 parts of vinegar tortoise shell, 10-15 parts of prepared rehmannia root, 6-20 parts of salt dodder seeds, 9-15 parts of wine polygonatum, 10-30 parts of kudzu root, 9-15 parts of knotweed, 6-20 parts of acanthopanax senticosus, 5-15 parts of salvia miltiorrhiza, 6-10 parts of roasted epimedium, and 3-10 parts of tangerine peel.

[0012] The selection of the above medicinal materials is the result of the inventor's exploration and summary based on the theory of traditional Chinese medicine after years of clinical experience, and has been analyzed, studied and verified. The prescription of the traditional Chinese medicine composition is concise and has good efficacy in improving ovarian function, increasing fertility and regulating bone metabolism.

[0013] In the prescription of the present invention, stir-fried tortoise plastron and prepared rehmannia root are the monarch drugs. Tortoise plastron tastes salty and slightly sweet, and is cool in nature. It belongs to the heart, liver, and kidney meridians. Its functions are to nourish yin and suppress yang, and nourish the heart with blood. Prepared rehmannia root tastes sweet and is slightly warm in nature. It belongs to the liver and kidney meridians. Its functions are to enrich blood and nourish yin. The combination of these two drugs has the effects of replenishing essence and tonifying the kidney, and nourishing blood and yin. Cuscuta chinensis and polygonatum sibiricum are the ministerial drugs together. Polygonatum sibiricum replenishes qi, nourishes yin, and benefits the kidney, assisting the monarch drugs to strengthen the function of replenishing the kidney and filling essence. "Those who are good at nourishing yin must seek yin from yang", so cuscuta chinensis and epimedium are added to tonify the kidney yang and enhance the effects of replenishing essence and nourishing blood. Pueraria lobata, polygonum cuspidatum, and acanthopanax senticosus are the assistant drugs. Acanthopanax senticosus benefits qi, strengthens the spleen, tonifies the kidney, and soothes the nerves. Pueraria lobata relieves muscle soreness and fever. Polygonum cuspidatum promotes blood circulation to remove blood stasis. The rest are the guiding drugs. Tangerine peel regulates qi and strengthens the spleen. Salvia miltiorrhiza nourishes blood, promotes blood circulation, and regulates menstruation, making the prescription tonifying without stagnation. The combination of all these drugs exerts the effects of replenishing essence and tonifying the kidney, dredging and regulating qi and blood, and is used to treat DOR of liver-kidney yin deficiency type. It makes the kidney qi sufficient, the heavenly water essence has a source, the thoroughfare and conception vessels are unobstructed, and the blood sea is full, thus effectively improving ovarian reserve function.

[0014] Further, the traditional Chinese medicine composition comprises the following raw materials in parts by weight: stir-fried tortoise plastron with vinegar 10 parts, prepared rehmannia root 12 parts, cuscuta chinensis with salt 6 parts, polygonatum sibiricum with wine 9 parts, pueraria lobata 20 parts, polygonum cuspidatum 10 parts, acanthopanax senticosus 6 parts, salvia miltiorrhiza 10 parts, epimedium with honey 9 parts, tangerine peel 6 parts.

[0015] Further, the traditional Chinese medicine composition comprises the following raw materials in parts by weight: stir-fried tortoise plastron with vinegar 15 parts, prepared rehmannia root 15 parts, cuscuta chinensis with salt 10 parts, polygonatum sibiricum with wine 15 parts, pueraria lobata 10 parts, polygonum cuspidatum 9 parts, acanthopanax senticosus 20 parts, salvia miltiorrhiza 5 parts, epimedium with honey 6 parts, tangerine peel 9 parts.

[0016] Further, the traditional Chinese medicine composition comprises the following raw materials in parts by weight: stir-fried tortoise plastron with vinegar 30 parts, prepared rehmannia root 10 parts, cuscuta chinensis with salt 20 parts, polygonatum sibiricum with wine 10 parts, pueraria lobata 30 parts, polygonum cuspidatum 12 parts, acanthopanax senticosus 15 parts, salvia miltiorrhiza 15 parts, epimedium with honey 10 parts, tangerine peel 10 parts.

[0017] Further, the traditional Chinese medicine composition comprises the following raw materials in parts by weight: stir-fried tortoise plastron with vinegar 30 parts, prepared rehmannia root 15 parts, cuscuta chinensis with salt 20 parts, polygonatum sibiricum with wine 10 parts, pueraria lobata 30 parts, polygonum cuspidatum 10 parts, acanthopanax senticosus 10 parts, salvia miltiorrhiza 15 parts, epimedium with honey 10 parts, tangerine peel 10 parts.

[0018] In the second aspect, the present invention provides a pharmaceutical preparation, which comprises the above traditional Chinese medicine composition.

[0019] Further, the pharmaceutical preparation comprises a pharmaceutically acceptable carrier or excipient.

[0020] Further, the pharmaceutically acceptable carrier or excipient is selected from at least one of a solvent, a diluent, a disintegrant, a precipitation inhibitor, a surfactant, a glidant, a binder, a lubricant, a dispersant, a suspending agent, an isotonic agent, a thickening agent, an emulsifier, a preservative, a stabilizer, a hydrating agent, an emulsification accelerator, a buffer, an absorbent, a coloring agent, a flavoring agent, a sweetening agent, an ion exchanger, a mold release agent, a coating agent, a flavor correcting agent or an antioxidant.

[0021] Further, the dosage form of the pharmaceutical preparation is a decoction, a medicinal broth, a granule, a pill, a capsule, a tablet, a powder or an oral liquid.

[0022] In a third aspect, the present invention provides a use of the above-mentioned traditional Chinese medicine composition in the preparation of a drug for treating ovarian reserve function decline and related diseases.

[0023] Further, the diseases related to ovarian reserve function decline include infertility.

[0024] Further, the ovarian reserve function decline is the ovarian reserve function decline of the liver-kidney yin deficiency type.

[0025] Further, the above-mentioned traditional Chinese medicine composition is used in combination with other estrogen drugs. Preferably, the other estrogen drugs are selected from Femoston and Climen.

[0026] Further, the daily dosage of the above-mentioned traditional Chinese medicine composition can be adjusted according to the needs of the condition. For example, the traditional Chinese medicine composition granule is administered to a patient, the packaging specification is 9 g / bag, 1 bag each time, 2 times a day.

[0027] In a fourth aspect, the present invention provides a preparation method of the above-mentioned traditional Chinese medicine composition, comprising the following steps:

[0028] Vinegar-processed turtle shell is pre-decocted with 8 - 12 (for example, 10) times the amount of water of the prescription for 0.5 - 1 h (for example, 0.5 h), then the remaining nine raw medicinal materials are added, and 8 - 12 (for example, 10) times the amount of water of the whole prescription is added to decoct for 0.5 - 1.5 h (for example, 0.5 h), the medicinal liquid is filtered out, then 6 - 10 (for example, 8) times the amount of water of the whole prescription is added to decoct for 0.5 - 1 h (for example, 0.5 h), the medicinal liquid is filtered out, the two decocted medicinal liquids are combined, and the combined medicinal liquid is concentrated under reduced pressure (≤75 °C, for example, 50 °C - 75 °C) to an extract with a relative density of 1.0 - 1.2 (for example, 1.08 ± 0.01) (25 °C), centrifuged or filtered, the supernatant is taken and dried to obtain the traditional Chinese medicine composition of the present invention.

[0029] Further, the traditional Chinese medicine composition of the present invention can be prepared into a pharmaceutical preparation containing the traditional Chinese medicine composition according to the conventional preparation methods in the art. The dosage form of the pharmaceutical preparation can be a decoction, a medicinal broth, a granule, a pill, a capsule, a tablet, a powder or an oral liquid.

[0030] In addition, unless otherwise specified, each raw material drug of the traditional Chinese medicine composition of the present invention can be obtained through commercial purchase. Any range described in the present invention includes the end values, any numerical value between the end values, and the traditional Chinese medicine composition of any sub-range constituted by any numerical value between the end values can achieve the purpose of treating decreased ovarian reserve function.

[0031] The beneficial effects of the present invention are as follows:

[0032] The traditional Chinese medicine composition of the present invention can effectively improve the ovarian function and sex hormone levels of patients with decreased ovarian reserve function, increase the number of their AFC, regulate the ovarian blood flow state, improve the menstrual conditions, traditional Chinese medicine syndromes and anxiety levels of patients, increase the clinical pregnancy rate, and has reliable safety. It realizes the targeted treatment of traditional Chinese medicine for decreased ovarian reserve function, especially for decreased ovarian reserve function of the liver and kidney yin deficiency type, filling the blank of drugs for treating decreased ovarian reserve function. Specific embodiments

[0033] To illustrate the present invention more clearly, the present invention will be further described below in conjunction with preferred embodiments. Those skilled in the art should understand that the specific content described below is illustrative rather than restrictive, and should not be used to limit the protection scope of the present invention.

[0034] Medical term names and abbreviations used in the present invention:

[0035] Antral follicle count: AFC

[0036] Ovarian stromal blood flow resistance index: RI

[0037] Anti-Müllerian hormone: AMH

[0038] Follicle-stimulating hormone: FSH

[0039] Luteinizing hormone: LH

[0040] Estradiol: E2

[0041] Pulsatility index: PI

[0042] End-diastolic blood flow velocity of ovarian artery: EDV

[0043] Peak systolic blood flow velocity of ovarian artery: PSV

[0044] Preparation of the traditional Chinese medicine composition in Examples 1 to 4

[0045]

[0046]

[0047] Weigh the above ten raw medicinal materials respectively according to the prescription ratios in Examples 1 to 4. The stir-fried tortoise plastron with vinegar is pre-boiled with 10 times the amount of water in the prescription for 0.5 h, and then the remaining nine raw medicinal materials are added. Then add 10 times the amount of water in the whole prescription and boil for 0.5 h. Filter out the medicinal liquid, add 8 times the amount of water in the whole prescription and boil for 0.5 h. Filter out the medicinal liquid, combine the two medicinal liquids, and concentrate the combined medicinal liquid under reduced pressure (70 °C) to an extract with a relative density of 1.08 ± 0.01 (25 °C). Centrifuge or filter, take the supernatant, and dry it to obtain the traditional Chinese medicine composition of the present invention.

[0048] Preparation of the granule of the traditional Chinese medicine composition in Examples 5 to 8

[0049] Weigh the above ten raw medicinal materials respectively according to the prescription ratios in Examples 1 to 4. The stir-fried tortoise plastron with vinegar is pre-boiled with 10 times the amount of water in the prescription for 0.5 h, and then the remaining nine raw medicinal materials are added. Then add 10 times the amount of water in the prescription and boil for 0.5 h. Filter out the medicinal liquid, add 8 times the amount of water in the prescription and boil for 0.5 h. Filter out the medicinal liquid, combine the two medicinal liquids, and concentrate the combined medicinal liquid under reduced pressure (70 °C) to an extract with a relative density of 1.08 ± 0.01 (25 °C). Centrifuge or filter, take the supernatant, dry it to obtain dry extract powder, add 2 g of sucralose, 5 g of magnesium stearate, and an appropriate amount of lactose to 1000 g, and granulate by dry granulation method, and package to obtain (for example, the packaging specification is 9 g / bag).

[0050] Preparation of the decoction of the traditional Chinese medicine composition in Examples 9 to 12

[0051] Weigh the above ten raw medicinal materials respectively according to the prescription ratios in Examples 1 to 4, add 10 times the amount of water and boil twice, boiling for 1.5 hours and 1 hour respectively, combine the decoction liquids, and filter to obtain.

[0052] The above examples list the preparation of the granule and decoction of the traditional Chinese medicine composition of the present invention. The traditional Chinese medicine composition of the present invention can also be prepared into the decoction, pill, capsule, tablet, powder or oral liquid of the traditional Chinese medicine composition of the present invention based on the conventional preparation methods in the art, and all are within the protection scope of the present invention.

[0053] Experimental part

[0054] Experimental Example 1 Efficacy test of the traditional Chinese medicine composition of the present invention in treating ovarian reserve function decline of liver-kidney yin deficiency type

[0055] 1 Data and methods

[0056] 1.1 General data

[0057] Sixty patients with DOR of liver-kidney yin deficiency type who visited the gynecology department of Xiyuan Hospital, China Academy of Chinese Medical Sciences from September 2018 to December 2020 were included. Among them, there were 30 patients with decreased ovarian reserve function and 30 patients with premature ovarian failure. The research subjects were divided into the observation group and the control group according to the ratio of 2:1. This study has obtained the approval of the Ethics Committee of Xiyuan Hospital, China Academy of Chinese Medical Sciences (Ethical Approval Number: Approval Number 2016XLA154).

[0058] 1.2 Diagnostic and Syndrome Differentiation Criteria

[0059] 1.2.1 Western Medicine Diagnostic Criteria

[0060] Formulated with reference to the "Expert Consensus on Poor Ovarian Response" of the Reproductive Medicine Branch of the Chinese Medical Association in 2015, the "Expert Consensus on Hormone Replacement Therapy for Premature Ovarian Insufficiency" in 2016, the "Chinese Expert Consensus on the Clinical Diagnosis and Treatment of Premature Ovarian Insufficiency" in 2017, and relevant literature:

[0061] 1) Decreased ovarian reserve function

[0062] a. Clinical manifestations: menstrual disorders, or amenorrhea;

[0063] b. Serum follicle-stimulating hormone (FSH): 10 mIU / mL < FSH < 40 mIU / mL;

[0064] c. Anti-Müllerian Hormone (AMH) < 1.1 ng / mL;

[0065] d. Bilateral ovarian basal antral follicle count (AFC) < 6;

[0066] Among them, b. - d. are all measured on the 2nd - 4th day of the menstrual cycle. For amenorrheic patients, pregnancy needs to be excluded. If the endometrium is less than 6 mm by ultrasound examination and there are no follicles larger than 1 cm in both ovaries, it can be directly detected;

[0067] Any 2 items of a. - d. are satisfied.

[0068] 2) Premature ovarian failure

[0069] a. Age < 40 years old;

[0070] b. Clinical manifestations: oligomenorrhea or amenorrhea for at least 4 months or more;

[0071] c. Serum FSH ≥ 40 mIU / mL for 2 times (the interval between the 2 examinations is more than 1 month);

[0072] On the 2nd to 4th day of menstruation, serum hormone SH, serum luteinizing hormone (LH), serum estradiol (E2) and AMH levels are measured. For amenorrhea, pregnancy is excluded, and ultrasound examination shows that the endometrium is less than 6mm and there are no follicles larger than 1cm in both ovaries, and serum hormone levels are directly measured.

[0073] 1.2.2 Diagnostic criteria for TCM syndrome types

[0074] The diagnostic criteria for liver-kidney yin deficiency syndrome were formulated with reference to the "Guidelines for Clinical Research of New Chinese Medicines (Trial)" issued by the State Food and Drug Administration in 2002, the textbooks "Diagnosis of Traditional Chinese Medicine" and "Gynecology of Traditional Chinese Medicine" (seventh edition textbook):

[0075] 1) Main symptoms: abnormal menstruation, such as early or late menstruation, amenorrhea, or scanty menstruation;

[0076] 2) Secondary symptoms: a. Weakness of waist and knees; b. Heat in the five parts of the body; c. Hot flashes, sweating or bone steaming fever; d. Dizziness or tinnitus; e. Dry throat and dry mouth; f. Dry eyes or blurred vision; g. Irritability or irritability; h. Insomnia, dreaminess or forgetfulness; i. Red tongue with little coating; j. Thready and rapid pulse;

[0077] As long as the main symptoms are met, 3 of the secondary symptoms will suffice.

[0078] 1.3 Inclusion criteria

[0079] Patients aged between 20 and 39 years old who meet the above-mentioned Western medical diagnostic criteria and TCM syndrome diagnostic criteria for decreased ovarian reserve function and premature ovarian failure, and who have no desire for pregnancy in the past three menstrual cycles, and who voluntarily participate and sign the informed consent form, can be included in the trial.

[0080] 1.4 Exclusion criteria

[0081] 1) Patients with congenital genital organ developmental abnormalities, or primary or secondary amenorrhea caused by acquired organic lesions and injuries;

[0082] 2) Patients with abnormal liver and kidney function and related endocrine diseases, such as diabetes;

[0083] 3) Patients with a history of ovarian surgery;

[0084] 4) Patients with serious primary diseases of cardiovascular, liver, kidney and hematopoietic system, and mental illness;

[0085] 5) Patients with menstrual disorders caused by uterine lesions, ovarian cysts and other gynecological diseases;

[0086] 6) Lactating women;

[0087] 7) Subjects with allergic constitution, who are allergic to multiple drugs or known to be allergic to the components of this medicine.

[0088] 1.5 Withdrawal Criteria

[0089] 1) Subjects who withdraw on their own initiative;

[0090] 2) Subjects who experience adverse events during the trial and are unable to continue participating in the trial;

[0091] 3) Lost to follow-up subjects

[0092] 1.6 Exclusion Criteria

[0093] 1) Subjects who do not take the treatment drugs as prescribed during the trial, affecting the determination of curative effect;

[0094] 2) Subjects who use other drugs that may interfere with the evaluation of curative effect during the trial, making it impossible to evaluate normally;

[0095] 3) Poor compliance, unable to complete the entire protocol;

[0096] 4) Incomplete data, affecting the evaluation of curative effect.

[0097] 1.7 Treatment Regimen

[0098] All subjects in the observation group took Baoluan Ankun Prescription (the traditional Chinese medicine composition in Example 4). It was taken starting from the 5th day of the menstrual cycle, 1 dose per day, taken warm in two divided doses in the morning and evening, and stopped taking during menstruation.

[0099] The control group took the combined package of Estradiol Valerate Tablets / Estradiol Cypionate Tablets (Bayer Healthcare Pharmaceuticals Inc., batch number: 20170814, specification: 21 tablets / box. The first 11 tablets (white tablets) each contain 2 mg of estradiol valerate, and the last 10 tablets (orange-red tablets) each contain 2 mg of estradiol valerate + 1 mg of cyproterone acetate. Administration method: For those with spontaneous menstruation, start taking the white tablets continuously from the 5th day of the menstrual cycle, 1 tablet / d. After 11 days, take the orange-red tablets, 1 tablet / d. Stop taking after 10 days, and menstruation will occur about 1 week later. Continue the previous method in the next cycle.

[0100] The treatment cycles for both the observation group and the control group included 3 menstrual cycles. Strict contraception was required during the treatment period. For all enrolled cases without spontaneous menstruation, pregnancy was excluded first. The endometrial thickness was detected by ultrasound. For those with an endometrial thickness < 8 mm, treatment started on the same day; for those who did not have menstruation 1 week after drug withdrawal, pregnancy was excluded, and medication for the next cycle could start on the same day. For those with an endometrial thickness ≥ 8 mm, intramuscular injection of progesterone injection 20 mg × 3 d was given first and then the drug was withdrawn. Treatment started on the 5th day of vaginal bleeding. During the treatment period, if there were cases without spontaneous menstruation, pregnancy was excluded first. The endometrial thickness was detected by ultrasound. For those with an endometrial thickness < 8 mm, treatment started on the same day; for those who did not have menstruation 1 week after drug withdrawal, pregnancy was excluded, and medication for the next cycle could start on the same day. For those with an endometrial thickness ≥ 8 mm, intramuscular injection of progesterone injection 20 mg × 3 d was given first and then the drug was withdrawn. Treatment started on the 5th day of vaginal bleeding.

[0101] 1.8 Observation indicators

[0102] The ovarian function was evaluated by the levels of serum AMH, FSH, LH, and E2 on the 2nd - 4th days of menstruation before and after 3 menstrual cycles of treatment. At the same time, the number of antral follicles, ovarian volume, and stromal blood flow resistance index (RI) (calculated by ultrasound), TCM syndrome scores, and changes in curative effects were observed on the 2nd - 4th days of menstruation before and after 3 menstrual cycles of treatment. The TCM syndrome scores were obtained by the researchers asking the patients about their conditions and filling out scales, and the total weighted symptom scores of the patients before and after 3 menstrual cycles of treatment were calculated and compared as the TCM syndrome scores. See Table 1.

[0103] Table 1 TCM syndrome scores

[0104]

[0105] 1.9 Statistical methods

[0106] SPSS 26.0 statistical software was used for data analysis. Among them, count data were expressed as rate or constituent ratio, and the chi - square test was used. Measurement data that followed a normal or approximately normal distribution were expressed as mean ± standard deviation (x ± s). Independent - sample t - test was used for between - group comparison, and paired t - test was used for within - group comparison; because the curative effect indicators did not conform to a normal distribution, the hypothesis test for between - group differences used the non - parametric rank - sum test, and the hypothesis test for within - group pre - and post - differences used the paired rank - sum test. The test level was two - sided α = 0.05, and P < 0.05 was considered statistically significant.

[0107] 2 Results

[0108] 2.1 Age and disease course

[0109] In the observation group, there were 40 females, aged 26 - 39 years, with an average age of (35.00 ± 3.62) years, and the disease course was 6 - 72 months, with an average disease course of (21.55 ± 19.38) months; in the control group, there were 20 females, aged 25 - 39 years, with an average age of (32.45 ± 4.91) years, and the disease course was 6 - 60 months, with an average disease course of (21.40 ± 17.52) months. The general data of the two groups of patients were analyzed statistically, and the differences were not statistically significant (P > 0.05), showing comparability.

[0110] 2.2 Changes in serological indicators

[0111] 2.2.1 Comparison of serological indicator levels in two groups of patients After treatment, the levels of SH and LH in the observation group were lower than those before treatment (both P < 0.05). There were no statistically significant differences in the levels of E2 and AMH (both P > 0.05). When comparing the serological indicators of the control group before and after treatment, there were no statistically significant differences (both P > 0.05). When comparing between the two groups, there were no statistically significant differences in the serological indicators (both P > 0.05). See Table 2.

[0112] Table 2 Comparison of serological indicator levels in two groups of patients

[0113]

[0114] Note: Compared with before treatment, * P < 0.05

[0115] 2.2.2 Changes in serological indicator levels in two groups of patients with decreased ovarian reserve function After treatment, the levels of SH and LH in the observation group decreased (both P < 0.05). There were no statistically significant differences in the serological indicators of the control group before and after treatment (both P > 0.05). When comparing the serological indicators between the two groups, there were no statistically significant differences (both P > 0.05). See Table 3.

[0116] Table 3 Comparison of serological indicator levels in two groups of patients with decreased ovarian reserve function

[0117]

[0118] Note: Compared with before treatment, * P < 0.05

[0119] 2.2.3 Changes in serological index levels in two groups of patients with premature ovarian failure After treatment, the level of $H in the observation group decreased compared with that before treatment (P<0.05), and there were no significant differences in the other serological indexes (all P>0.05). When comparing the serological indexes of the control group before and after treatment, there were no significant differences (all P>0.05). When comparing between groups, there were no significant differences in each serological index between the observation group and the control group before and after treatment (all P>0.05), but the improvement of the laboratory indexes in the observation group was more obvious, which had certain clinical significance. See Table 4.

[0120] Table 4 Comparison of serological index levels in two groups of patients with premature ovarian failure

[0121]

[0122] Note: Compared with before treatment, * P<0.05

[0123] 2.3 Changes in ultrasonic indexes

[0124] In patients with decreased ovarian reserve function and patients with premature ovarian failure, the changes in AFC, the average volume of bilateral ovaries, and the ovarian stromal blood flow resistance index (Resistance In-dx, R) were compared between the two groups before and after treatment. The results showed that there were no significant differences (all P>0.05), and there were no significant differences in each ultrasonic index between the two groups after treatment (all P>0.05). See Tables 5-7.

[0125] Table 5 Overall comparison of ultrasonic indexes in two groups of patients

[0126]

[0127] Table 6 Comparison of ultrasonic indexes in patients with decreased ovarian reserve function

[0128]

[0129] Table 7 Comparison of ultrasonic indexes in patients with premature ovarian failure

[0130]

[0131] 2.4 Comparison of TCM syndrome scores

[0132] 2.4.1 Comparison of TCM syndrome scores in two groups of patients After three menstrual cycles of treatment, the TCM syndrome score of the control group was lower than that before treatment, and the difference was statistically significant (P<0.05). The TCM syndrome score of the observation group was also significantly lower than that before treatment, and the difference was statistically significant (P<0.01), and the decline was greater than that of the control group (P<0.01). See Table 8.

[0133] Table 8 Comparison of Traditional Chinese Medicine Syndrome Scores between Two Groups of Patients ( points)

[0134]

[0135] Note: Compared with before treatment, * P < 0.05, ＊＊ P < 0.01; compared with the control group, ΔΔ P < 0.01

[0136] 2.4.2 Comparison of Traditional Chinese Medicine Syndrome Scores between Two Groups of Patients with Declined Ovarian Reserve Function After treatment, the Traditional Chinese Medicine syndrome score of the observation group was significantly lower than that before treatment, with a statistically significant difference (P < 0.01), while there was no statistically significant difference in the Traditional Chinese Medicine syndrome score of the control group (P > 0.05). In the comparison between groups, there was no statistically significant difference in the Traditional Chinese Medicine syndrome scores of the two groups after treatment (P > 0.05). See Table 9.

[0137] Table 9 Comparison of Traditional Chinese Medicine Syndrome Scores between Two Groups of Patients with Declined Ovarian Reserve Function ( points)

[0138]

[0139] Note: Compared with before treatment, ＊＊ P < 0.01

[0140] 2.4.3 Comparison of Traditional Chinese Medicine Syndrome Scores between Two Groups of Patients with Premature Ovarian Failure After treatment, the Traditional Chinese Medicine syndrome score of the control group was lower than that before treatment, with a statistically significant difference (P < 0.05), while the Traditional Chinese Medicine syndrome score of the observation group was significantly lower than that before treatment, with a statistically significant difference (P < 0.01), and it was superior to the control group (P < 0.01). See Table 10.

[0141] Table 10 Comparison of Traditional Chinese Medicine Syndrome Scores between Two Groups of Patients with Premature Ovarian Failure ( points)

[0142]

[0143] Note: Compared with before treatment, * P < 0.05, ＊＊ P < 0.01; compared with the control group, ΔΔ P < 0.01

[0144] 2.5 Comparison of Therapeutic Effects of Traditional Chinese Medicine Syndromes

[0145] 2.5.1 Comparison of Therapeutic Effects of Traditional Chinese Medicine Syndromes between Two Groups of Patients After treatment, the therapeutic effect of Traditional Chinese Medicine syndromes in the observation group was significantly better than that in the control group, with a statistically significant difference (P < 0.01). See Table 11.

[0146] Table 11 Overall comparison of the efficacy of TCM syndromes in the two groups of patients [cases (%)]

[0147]

[0148] Note: Compared with the control group, ＊＊ P<0.01

[0149] 2.5.2 Comparison of the efficacy of TCM syndromes in 2 groups of patients with decreased ovarian reserve function After treatment, the efficacy of TCM syndromes in the observation group with decreased ovarian reserve function was better than that in the control group, and the difference was statistically significant (P<0.01). See Table 12.

[0150] Table 12 Comparison of TCM syndrome efficacy between two groups of patients with decreased ovarian reserve function [cases (%)]

[0151]

[0152] Note: Compared with the control group, ＊＊ P<0.01

[0153] 2.5.3 Comparison of the efficacy of TCM syndromes in 2 groups of patients with premature ovarian failure After treatment, the efficacy of TCM syndromes in the observation group of premature ovarian failure was better than that in the control group, and the difference was statistically significant (P<0.01). See Table 13.

[0154] Table 13 Comparison of TCM syndrome efficacy in patients with premature ovarian failure [cases (%)]

[0155]

[0156] Note: Compared with the control group, ＊＊ P<0.01

[0157] 2.6 Summary

[0158] In summary, the Chinese medicine composition of the present invention can significantly improve the clinical symptoms of patients with liver-kidney yin deficiency type DOR and regulate the level of sex hormones, and is worthy of clinical application and promotion.

[0159] Experimental Example 2 Efficacy and safety of the Chinese medicine composition of the present invention in treating liver-kidney yin deficiency type ovarian reserve dysfunction accompanied by anxiety

[0160] 1 Materials and methods

[0161] 1.1 General Information

[0162] A total of 118 patients with liver-kidney yin deficiency type DOR who visited the Department of Gynecology of Xiyuan Hospital and Wangjing Hospital of China Academy of Chinese Medical Sciences from April 2022 to April 2025 were included. This study has been approved by the Ethics Committee of Xiyuan Hospital of China Academy of Chinese Medical Sciences (Ethics Approval No.: Approval No. 2022XLA042).

[0163] 1.2 Diagnosis and syndrome differentiation criteria

[0164] 1.2.1 The diagnostic criteria for DOR refer to "Expert Consensus on the Clinical Diagnosis and Treatment of Ovarian Reserve Decline", "Integrated Traditional Chinese and Western Medicine Diagnosis and Treatment Guidelines for Premature Ovarian Insufficiency", "Management of Premature Ovarian Insufficiency in Women", "Expert Consensus on Hormone Replacement Therapy for Premature Ovarian Insufficiency", etc., and the following criteria are formulated:

[0165] (1) The age range is from 18 to 40 years old;

[0166] (2) The clinical manifestations are menstrual disorders or amenorrhea or accompanied by infertility;

[0167] (3) 10 U / L ≤ FSH < 25 U / L;

[0168] (4) 0.5 ≤ AMH < 1.1 ng / ml;

[0169] (5) AFC < 6. Among them, the sex hormone levels and ultrasound indicators are all basic levels, and are evaluated on the 2nd - 4th day of the menstrual cycle. If the patient has amenorrhea, after excluding pregnancy, if the endometrium ≤ 5 mm and the bilateral ovarian follicle diameters are both < 1 cm, the sex hormone and AFC determinations can be directly performed.

[0170] The above (1) and (2) are the necessary conditions for inclusion in the group, and the patient can be diagnosed if any one of (3) - (5) is met.

[0171] 1.2.2 Diagnostic criteria for anxiety state

[0172] Anxiety state is a group of symptom syndromes. When a patient has anxiety symptoms that affect daily life but do not meet the diagnostic criteria for the course of anxiety disorder, it can be diagnosed as anxiety state by a non - psychiatrist.

[0173] In this study, the Self - Rating Anxiety Scale (SAS) was used for evaluation. A total score below 50 points indicates normal, 50 - 59 points indicates that the patient has mild anxiety, 60 - 69 points indicates that the patient has moderate anxiety, and 70 points and above indicate that the patient has severe anxiety.

[0174] 1.2.3 Traditional Chinese medicine syndrome criteria

[0175] Taking the syndrome of liver - kidney yin deficiency as the research object: It is formulated according to "Diagnostic Criteria and Therapeutic Effects of Traditional Chinese Medicine Syndromes" and "Diagnosis and Treatment Guidelines for Common Diseases in Traditional Chinese Gynecology".

[0176] (1) Main symptoms: Abnormal menstruation, such as scanty menstruation, early menstruation, late menstruation or even amenorrhea, etc.

[0177] (2) Secondary symptoms: flushing of the cheekbones with feverish sensation, feverish sensation in the five centers (palms, soles and chest), restlessness and insomnia, dryness of the throat and thirst, vaginal dryness, soreness and weakness of the waist and knees, dizziness and tinnitus, feverish sweating, hectic fever, dull pain in the chest and hypochondrium, irritability and anger, dryness of both eyes or blurred vision, etc.

[0178] (3) Tongue and pulse: red tongue, little coating, thready and rapid pulse. The main symptoms are essential, and any 3 secondary symptoms are satisfied, and the diagnosis can be made by conforming to the tongue and pulse manifestations at the same time.

[0179] 1.3 Inclusion criteria

[0180] (1) Meet the western medicine diagnostic criteria of DOR.

[0181] (2) Meet the western medicine diagnostic criteria of mild and moderate anxiety states.

[0182] (3) The TCM syndrome differentiation conforms to the syndrome of deficiency of both liver and kidney yin.

[0183] (4) The participants actively sign the informed consent form.

[0184] 1.4 Exclusion criteria

[0185] (1) Suffering from congenital dysplasia of reproductive organs or amenorrhea caused by acquired organic lesions and injuries.

[0186] (2) Suffering from severe primary diseases such as cardiovascular, liver, kidney and hematopoietic systems, as well as diabetes patients.

[0187] (3) Having undergone bilateral or unilateral ovariectomy.

[0188] (4) Having been diagnosed or suspected of breast cancer, having been diagnosed or suspected of precancerous lesions or malignant tumors affected by sex hormones.

[0189] (5) Acute arterial thromboembolism, active venous thrombosis, history of thromboembolic diseases, or those with high risk factors for venous or arterial thrombosis.

[0190] (6) Having used hormones within the past 3 months.

[0191] (7) Those allergic to the ingredients of this medicine.

[0192] (8) Severe hypertriglyceridemia.

[0193] (9) Those with a clear diagnosis of mental illness or communication disorders in the past.

[0194] (10) Those who have taken anti-anxiety and antidepressant drugs recently.

[0195] (11) Those with poor compliance of the subjects, unable to accurately evaluate the curative effect, or those whose curative effect judgment is affected due to incomplete data shall be excluded.

[0196] 1.5 Withdrawal criteria

[0197] (1) Those who experience severe adverse reactions or develop complications during the treatment process, resulting in their inability to continue participating in the trial.

[0198] (2) Those who withdraw from the diagnosis and treatment midway due to personal reasons during the research period, or those who are unable to complete the research due to loss to follow-up.

[0199] (3) Those who request to withdraw due to personal reasons during the research process.

[0200] 1.6 Exclusion Criteria

[0201] (1) Those who are misdiagnosed or misenrolled because they do not meet the inclusion criteria.

[0202] (2) Those who do not comply with the treatment process, cannot take medicine on time, or take other drugs orally that affect the efficacy of the drugs in this project.

[0203] (3) Those who participate in other clinical projects simultaneously.

[0204] 1.7 Random Grouping

[0205] Through the random number table method, the subjects are included in the experimental group and the control group in the same proportion.

[0206] 1.8 Blinding Design and Emergency Unblinding

[0207] This study adopts a two-level blinding method. First, the case serial numbers are blinded so that the researchers can allocate the case serial numbers without knowing the situation. Second, the outer packages of the drugs are blinded so that the outer packages of the two groups of drugs are kept consistent, and neither the researchers nor the subjects know the drug grouping to exclude the influence of subjective factors and ensure the rigor of the trial. During the research process, the researchers and the subjects sign an informed consent to protect the rights and interests of both parties.

[0208] During the trial process, data should be collected in a timely manner and unblinding should be carried out before statistical analysis. If a major adverse event occurs during the trial, in order to protect the rights and interests of the subjects, emergency unblinding can be implemented. When implementing emergency unblinding, multiple parties should participate together, such as the sponsor monitor, the project leader, the researcher, etc., and detailed records should be made, including the reasons for unblinding, the time of unblinding, and the location of unblinding. This subject should stop participating in the clinical research at this time, be regarded as dropped out, and its efficacy indicators will not be included in the final efficacy evaluation, but its safety indicators should be fully analyzed, and relevant adverse reactions and complications should be treated in a timely manner to protect the rights and interests of the subjects.

[0209] 1.9 Treatment Plan

[0210] 1.9.1 Test Drugs

[0211] Experimental Group:

[0212] Baoluan Ankun Granules: The granule of the traditional Chinese medicine composition in Example 8, 9 g / bag;

[0213] Femoston placebo: Purchased from Beijing Kangrentang Pharmaceutical Co., Ltd., specifications: 14 white tablets and 14 grey tablets, both are starch-containing tablets, used as placebo without any therapeutic effect.

[0214] Control group:

[0215] Baoluan Ankun Granule placebo: Purchased from Beijing Kangrentang Pharmaceutical Co., Ltd., at 5% of the original formula (Baoluan Ankun Granule) dose;

[0216] Femoston: Purchased from the pharmacy of Xiyuan Hospital, China Academy of Chinese Medical Sciences, specifications: the white tablets are estradiol tablets 1mg, 14 tablets, the grey tablets are estradiol 1mg + dydrogesterone 10mg, 14 tablets, a total of 28 tablets / box.

[0217] 1.9.2 Medication time

[0218] For patients with spontaneous menstruation, start taking the medicine on the fifth day of the menstrual cycle. For patients without spontaneous menstruation, pregnancy should be excluded first. If the ultrasound shows that the endometrial thickness (En) is less than 8 mm and there are no follicles larger than 1 cm in both ovaries, treatment can start on the same day; if En ≥ 8 mm, intramuscular injection of progesterone injection is required, with a dose of 20 mg / day for 3 consecutive days and then stop taking the medicine, and start treatment on the fifth day of vaginal bleeding.

[0219] 1.9.3 Medication method

[0220] Experimental group: Start taking Baoluan Ankun Granule and Femoston placebo on the fifth day of menstruation. Take 1 bag of Baoluan Ankun Granule twice a day; take Femoston placebo once a day, 1 tablet each time, first take the white tablets for 14 days, and then take the grey tablets for 14 days. Take continuously for 84 days.

[0221] Control group: Take Femoston and Baoluan Ankun Granule placebo orally on the fifth day of menstruation. The Baoluan Ankun Granule placebo is 9g / bag, take 1 bag twice a day; take Femoston once a day, 1 tablet each time, first take the white tablets for 14 days, and then take the grey tablets for 14 days, and take the medicine continuously for 84 days.

[0222] 1.10 Observation indicators

[0223] (1) General clinical data. Include patient age, course of disease, BMI, marriage and childbearing history, past history, etc. Record once within 7 days before enrollment and once within 7 days after leaving the group.

[0224] (2) Efficacy indicators: Serum AMH value, SAS anxiety scale, sex hormone indicators, ultrasound indicators, traditional Chinese medicine syndrome scoring scale, other scales, comprehensive efficacy determination.

[0225] (3) Safety indicators: Record the adverse reactions of patients during medication, and conduct blood routine, urine routine, liver function (ALT, AST), kidney function (BUN, Cr), and electrocardiogram examinations. All examinations need to be completed once within 7 days before enrollment and within 7 days after withdrawal.

[0226] 1.11 Treatment of adverse reactions

[0227] Observe whether there are any adverse reactions in the two groups of patients during the trial. Once any adverse reaction occurs in a patient, the researcher should make timely treatment, analyze the cause of its occurrence, report to the research principal and the ethics committee, and take corresponding protective measures.

[0228] 1.12 Statistical methods

[0229] Use SPSS 27.0 software to analyze the data. Measurement data are first subjected to a normality test. For measurement data that conform to a normal distribution, further perform a homogeneity of variance analysis. If the variances are homogeneous, an independent samples t-test is used for between-group comparisons, and a paired samples t-test is used for within-group pre- and post-data comparisons; if the variances are not homogeneous, an independent samples t-test (adjusting the degrees of freedom) is used for between-group comparisons, and a paired samples t-test (adjusting the degrees of freedom) is used for within-group pre- and post-data comparisons. For measurement data that do not conform to a normal distribution, the Mann-Whitney U test is used for between-group comparisons, and the Wilcoxon test is used for within-group pre- and post-data comparisons. Count data are expressed as rates or constituent ratios and are analyzed using the chi-square test (X 2 test). All tests use a two-sided significance level of α = 0.05. When the P value is less than 0.05, the difference is considered statistically significant.

[0230] 2 Results

[0231] 2.1 Completion of clinical trials

[0232] This study was conducted in the Department of Gynecology, Xiyuan Hospital, China Academy of Chinese Medical Sciences from April 2022 to April 2025. A total of 118 cases were randomly enrolled, with 59 cases in the experimental group and 59 cases in the control group. 101 subjects completed 3 menstrual cycle treatments, with 48 cases in the experimental group and 53 cases in the control group. In addition, 6 subjects were still under treatment (4 cases in the experimental group and 2 cases in the control group), and 4 subjects became pregnant during the medication process (2 cases in the experimental group and 2 cases in the control group), so they were not included in the statistical analysis. There were 7 dropout cases (4 cases in the experimental group and 3 cases in the control group).

[0233] 2.2 Baseline conditions

[0234] 2.2.1 Age and disease duration

[0235] The age distribution of the experimental group was 26 - 40 years old, with an average age of 34.75 ± 4.33 years. The age distribution of the control group was 22 - 40 years old, with an average age of 34.96 ± 4.32 years. There was no statistically significant difference in age between the two groups (P > 0.05). The disease course distribution of the experimental group was 0.25 - 8 years, with an average disease course of 2.20 ± 1.64 years. The disease course distribution of the control group was 0.17 - 5 years, with an average disease course of 1.67 ± 1.15 years. There was no statistically significant difference in disease course between the two groups (P > 0.05).

[0236] 2.2.2 Gravidity and parity

[0237] The gravidity distribution of the experimental group was 0 - 5 times, with an average gravidity of 1.33 ± 1.37 times. The gravidity distribution of the control group was 0 - 5 times, with an average gravidity of 1.19 ± 1.35 times. There was no statistically significant difference in gravidity between the two groups (P > 0.05). The parity distribution of the experimental group was 0 - 2 times, with an average parity of 0.67 ± 0.72 times. The parity distribution of the control group was 0 - 2 times, with an average parity of 0.58 ± 0.69 times. There was no statistically significant difference in parity between the two groups (P > 0.05).

[0238] 2.3 Efficacy analysis

[0239] 2.3.1 Comparison of AMH between the two groups before and after treatment

[0240] Among the 118 patients randomly enrolled, 101 patients were eligible for AMH comparison before and after treatment, including 48 in the experimental group and 53 in the control group. Intra-group comparison by wilconxon test showed that AMH in the experimental group increased significantly after treatment, P = 0.001 < 0.05, with statistically significant difference; AMH in the control group showed a downward trend after treatment, P = 0.884 > 0.05, with no statistically significant difference, as shown in Table 14. Inter-group comparison by Mann-Whitney U showed that the increasing trend of AMH in the experimental group was higher than that in the control group, P = 0.011 < 0.05, with statistically significant difference, as shown in Table 15.

[0241] Table 14 Intra-group comparison of AMH levels between the experimental group and the control group( ng / ml)

[0242]

[0243] Table 15 Inter-group comparison of AMH levels between the experimental group and the control group( ng / ml)

[0244]

[0245] 2.3.2 Comparison of sex hormones between the two groups before and after treatment

[0246] Among the 118 patients randomly assigned, 101 patients could have their FSH, LH, and E2 compared before and after treatment, including 48 patients in the experimental group and 53 patients in the control group. Intragroup comparison by the wilconxon test showed that after treatment, FSH decreased in the experimental group, with P = 0.015 < 0.05, indicating a statistically significant difference; there was a downward trend in FSH after treatment in the control group, with P = 0.743 > 0.05, indicating no statistically significant difference. There was a downward trend in LH after treatment in the experimental group, with P = 0.701 > 0.05, indicating no statistically significant difference; there was an upward trend in LH after treatment in the control group, with P = 0.270 > 0.05, indicating no statistically significant difference. There was an upward trend in E2 after treatment in the experimental group, with P = 0.176 > 0.05, indicating no statistically significant difference; E2 increased after treatment in the control group, with P = 0.022 < 0.05, indicating a statistically significant difference, as shown in Tables 16 - 18.

[0247] Table 16 Intragroup comparison of FSH levels between the experimental group and the control group ( uIU / mL)

[0248]

[0249] Table 17 Intragroup comparison of LH levels between the experimental group and the control group ( uIU / mL)

[0250]

[0251] Table 18 Intragroup comparison of E2 levels between the experimental group and the control group ( pmol / L)

[0252]

[0253] Intergroup comparison by the Mann-Whitney U test showed that the decreasing trend of FSH in the experimental group was lower than that in the control group, with P = 0.096 > 0.05, indicating no statistically significant difference. The decreasing trend of LH in the experimental group was lower than that in the control group, with P = 0.277 > 0.05, indicating no statistically significant difference. The increasing trend of E2 in the experimental group was lower than that in the control group, with P = 0.589 > 0.05, indicating no statistically significant difference, as shown in Tables 19 - 21.

[0254] Table 19 Intergroup comparison of FSH levels between the experimental group and the control group ( uIU / mL)

[0255]

[0256] Table 20 Intergroup comparison of LH levels between the experimental group and the control group ( uIU / mL)

[0257]

[0258] Comparison of E2 levels between the experimental group and the control group before and after treatment( (pmol / L)

[0259]

[0260]

[0261] 2.3.3 Comparison of AFC between the two groups before and after treatment

[0262] Among the 118 patients randomly enrolled, 101 patients could have their AFC compared before and after treatment, including 48 in the experimental group and 53 in the control group. Intra-group comparison by wilconxon test showed that there was an upward trend in AFC after treatment in the experimental group, with P = 1.128 > 0.05, and the difference was not statistically significant; there was an upward trend in AFC after treatment in the control group, with P = 1.134 > 0.05, and the difference was not statistically significant, as shown in Table 22.

[0263] Table 22 Intra-group comparison of AFC levels between the experimental group and the control group( (units)

[0264]

[0265] Inter-group comparison by Mann-Whitney U showed that the upward trend of AFC in the experimental group was lower than that in the control group, with P = 0.975 > 0.05, and the difference was not statistically significant, as shown in Table 23.

[0266] Table 23 Inter-group comparison of AFC levels between the experimental group and the control group( (units)

[0267]

[0268] 2.3.4 Comparison of ovarian blood flow between the two groups before and after treatment

[0269] Among the 118 patients randomly assigned, 80 patients could have PSV compared before and after treatment, including 35 in the experimental group and 45 in the control group; 58 patients could have EDV compared before and after treatment, including 27 in the experimental group and 31 in the control group; 65 patients could have PI compared before and after treatment, including 30 in the experimental group and 35 in the control group; 86 patients could have RI compared before and after treatment, including 39 in the experimental group and 47 in the control group. Intra-group comparison by wilconxon test showed that PSV in the experimental group increased significantly after treatment, P = 0.021 < 0.05, with statistically significant difference; PSV in the control group had a downward trend after treatment, P = 0.488 > 0.05, with no statistically significant difference. EDV in the experimental group had an upward trend after treatment, P = 0.086 > 0.05, with no statistically significant difference; EDV in the control group had a downward trend after treatment, P = 0.561 > 0.05, with no statistically significant difference. PI in the experimental group had a downward trend after treatment, P = 0.307 > 0.05, with no statistically significant difference; PI in the control group had an upward trend after treatment, P = 0.638 > 0.05, with no statistically significant difference. RI in the experimental group had a downward trend after treatment, P = 0.064 > 0.05, with no statistically significant difference; RI in the control group had an upward trend after treatment, P = 0.088 > 0.05, with no statistically significant difference, as shown in Tables 24 - 27.

[0270] Table 24 Intra-group comparison of PSV levels between the experimental group and the control group ( cm / s)

[0271]

[0272]

[0273] Table 25 Intra-group comparison of EDV levels between the experimental group and the control group ( cm / s)

[0274]

[0275] Table 26 Intra-group comparison of PI levels between the experimental group and the control group

[0276]

[0277] Table 27 Intra-group comparison of RI levels between the experimental group and the control group

[0278]

[0279] Inter-group comparison by Mann-Whitney U showed that the increasing trend of PSV in the experimental group was better than that in the control group, with P = 0.016 < 0.05, and the difference was statistically significant. The increasing trend of EDV in the experimental group was better than that in the control group, with P = 0.046 < 0.05, and the difference was statistically significant. The increasing trend of PI in the experimental group was lower than that in the control group, with P = 0.678 > 0.05, and the difference was not statistically significant. The decreasing trend of RI in the experimental group was better than that in the control group, with P = 0.855 > 0.05, and the difference was not statistically significant, as shown in Tables 28 - 31.

[0280] Table 28 Inter-group comparison of PSV levels between the experimental group and the control group ( cm / s)

[0281]

[0282] Table 29 Inter-group comparison of EDV levels between the experimental group and the control group ( cm / s)

[0283]

[0284] Table 30 Inter-group comparison of PI levels between the experimental group and the control group

[0285]

[0286] Table 31 Inter-group comparison of RI levels between the experimental group and the control group

[0287]

[0288] 2.3.5 Comparison of the scoring scales between the two groups before and after treatment

[0289] Among the 120 patients randomly assigned, 101 patients could have their menstrual scores, traditional Chinese medicine syndrome scores, modified Kupperman scores, and HAMA scores compared before and after treatment. There were 48 patients in the experimental group and 53 patients in the control group. Intra-group comparison by Wilcoxon test showed that the menstrual score in the experimental group decreased significantly after treatment, P < 0.001, with statistically significant differences; the menstrual score in the control group decreased significantly after treatment, P < 0.001, with statistically significant differences. The traditional Chinese medicine syndrome score in the experimental group decreased significantly after treatment, P < 0.001, with statistically significant differences; the traditional Chinese medicine syndrome score in the control group decreased significantly after treatment, P < 0.001, with statistically significant differences. The modified Kupperman score in the experimental group decreased significantly after treatment, P < 0.001, with statistically significant differences; the modified Kupperman score in the control group decreased significantly after treatment, P < 0.001, with statistically significant differences. The HAMA score in the experimental group decreased significantly after treatment, P < 0.001, with statistically significant differences; the HAMA score in the control group decreased significantly after treatment, P < 0.001, with statistically significant differences, as shown in Tables 32 - 35.

[0290] Table 32 Intra-group comparison of menstrual scores between the experimental group and the control group ( points)

[0291]

[0292] Table 33 Intra-group comparison of traditional Chinese medicine syndrome scores between the experimental group and the control group ( points)

[0293]

[0294] Table 34 Intra-group comparison of modified Kupperman scores between the experimental group and the control group ( points)

[0295]

[0296]

[0297] Table 35 Intra-group comparison of HAMA scores between the experimental group and the control group ( points)

[0298]

[0299] Inter-group comparison by Mann-Whitney U showed that the decreasing trend of menstrual scores in the experimental group was better than that in the control group, with P = 0.421 > 0.05, and the difference was not statistically significant. The decreasing trend of traditional Chinese medicine syndrome scores in the experimental group was better than that in the control group, with P < 0.001, and the difference was statistically significant. The decreasing trend of the modified Kupperman scores in the experimental group was better than that in the control group, with P < 0.001, and the difference was statistically significant. The decreasing trend of HAMA scores in the experimental group was better than that in the control group, with P = 0.019 < 0.05, and the difference was statistically significant, as shown in Tables 36 - 39.

[0300] Table 36 Inter-group comparison of menstrual scores between the experimental group and the control group ( points)

[0301]

[0302] Table 37 Inter-group comparison of traditional Chinese medicine syndrome scores between the experimental group and the control group ( points)

[0303]

[0304] Table 38 Inter-group comparison of modified Kupperman scores between the experimental group and the control group ( points)

[0305]

[0306] Table 39 Inter-group comparison of HAMA scores between the experimental group and the control group ( points)

[0307]

[0308] 2.4 Comparison of safety indicators

[0309] There were no obvious abnormalities in the physical and chemical examinations (blood routine, urine routine, liver and kidney function, electrocardiogram) before and after treatment in the experimental group and the control group.

[0310] 2.6 Follow-up

[0311] The subjects who had left the group were followed up for 3 months to understand the pregnancy and adverse event conditions of the subjects: During the follow-up period, there were 5 pregnant patients in total, including 2 in the experimental group and 2 in the control group who became pregnant during treatment, and 1 in the experimental group who became pregnant after treatment. During the follow-up period, no adverse events were reported in both groups of patients.

[0312] 2.7 Summary

[0313] In summary, the traditional Chinese medicine composition of the present invention can effectively improve the ovarian function and sex hormone levels of patients with DOR (liver-kidney yin deficiency syndrome), increase the number of their AFC, regulate the ovarian blood flow state of patients with DOR (liver-kidney yin deficiency syndrome), improve the menstrual conditions, traditional Chinese medicine syndromes and anxiety levels of patients, improve their overall quality of life, and has good safety.

[0314] Experimental Example 3 Efficacy test of the traditional Chinese medicine composition of the present invention in the treatment of infertility related to diminished ovarian reserve of liver-kidney yin deficiency type

[0315] 1 Data and methods

[0316] 1.1 General information

[0317] A total of 68 patients with infertility related to DOR of liver-kidney yin deficiency type who visited the Department of Gynecology of Xiyuan Hospital, China Academy of Chinese Medical Sciences from August 2020 to December 2021 were included. This study has obtained the approval of the Ethics Committee of Xiyuan Hospital, China Academy of Chinese Medical Sciences (Ethical Approval No.: Approval No. 2018XLA063).

[0318] 1.2 Diagnostic and syndrome differentiation criteria

[0319] 1.2.1 Western medicine diagnostic criteria

[0320] Refer to the "Testing and interpreting measures of ovarian reserve: a committee opinion" of the American Society for Reproductive Medicine in 2020 and the "Diagnosis Guidelines for Infertility" of the Obstetrics and Gynecology Branch of the Chinese Medical Association in 2019: The following indicators were measured on the 2nd - 3rd day of menstruation (for those without spontaneous menstruation, after excluding pregnancy and with no follicles > 1 cm in both ovaries detected by B-ultrasound, directly measured): Diagnostic criteria for DOR:

[0321] (1) The total number of bilateral AFC ≤ 6;

[0322] (2) AMH ≤ 1.1 ng / mL;

[0323] (3) 39 U / L ≥ bFSH ≥ 10 U / L;

[0324] (4) E2 > 292.8 pmol / L (i.e., 80 pg / mL);

[0325] Two of the above four items are met.

[0326] Diagnostic criteria for infertility: The couple has normal sexual life and has not been pregnant for 1 year or more without contraception.

[0327] 1.2.2 Traditional Chinese medicine syndrome criteria

[0328] Refer to the "Terms of Traditional Chinese Medicine Clinical Diagnosis and Treatment" in 2020 and the textbook "Traditional Chinese Medicine Diagnosis" for formulation:

[0329] (1) Infertility, with or without menstrual disorders or amenorrhea;

[0330] (2) Dizziness and tinnitus;

[0331] (3) Hot flashes and sweating;

[0332] (4) Dry throat and thirst;

[0333] (5) Weakness and soreness of the waist and knees;

[0334] (6) Vaginal dryness;

[0335] (7) Insomnia and forgetfulness.

[0336] The tongue is red, the tongue coating is scanty, and the pulse is thready and rapid. Item (1) is essential, and if the other three items are present, the diagnosis can be made in combination with the tongue and pulse conditions.

[0337] 1.3 Inclusion criteria

[0338] (1) Those who meet the above Western medicine diagnostic criteria and traditional Chinese medicine syndrome differentiation criteria;

[0339] (2) Aged 20 - 40 years old;

[0340] (3) Have normal sexual life between husband and wife during the treatment period;

[0341] (4) At least one fallopian tube is unobstructed;

[0342] (5) Sign the informed consent form.

[0343] 1.4 Exclusion criteria

[0344] (1) Those with congenital reproductive organ dysplasia or defects, or infertility caused by genetic or immune factors;

[0345] (2) Those with other endocrine diseases causing infertility, such as polycystic ovary syndrome, hyperprolactinemia, etc.;

[0346] (3) Those with infertility caused by uterine and fallopian tube factors: fallopian tube obstruction, endometriosis, uterine hypoplasia, pelvic inflammation, genital tuberculosis, etc.;

[0347] (4) Abnormal semen routine in men: sperm concentration < 1.5×10 / mL, progressive motility sperm (PR) < 32%, and the sum of progressive motility sperm (PR) and non - progressive motility sperm (NP) < 40%;

[0348] (5) Those with severe primary diseases of the cardiovascular, liver, kidney, etc. and those with mental diseases who are unable to take care of themselves;

[0349] (6) Those who have taken traditional Chinese medicine and Western medicine related to improving ovarian reserve function within the past month, or those with contraindications to hormonal drug use;

[0350] (7) Subjects who are allergic to the investigational drug.

[0351] 1.5 Withdrawal Criteria

[0352] Patients who meet the inclusion and exclusion criteria, understand the entire process of the trial and sign the informed consent form, and withdraw without reason or fail to complete the treatment cycle according to the research implementation plan can be regarded as withdrawn cases. The reasons for withdrawal may be as follows:

[0353] (1) Those who experience adverse events such as skin itching, diarrhea and other allergic reactions after medication and are not suitable for continuing the clinical trial;

[0354] (2) Those who withdraw on their own initiative for personal reasons during the trial;

[0355] (3) Those who are lost to follow-up due to reasons such as epidemic prevention and control during treatment: For the withdrawn cases that drop out midway, relevant reasons should be actively inquired about and examinations should be completed as much as possible to analyze their treatment effects and safety.

[0356] 1.6 Exclusion Criteria

[0357] (1) Those who need to be terminated prematurely due to adverse reactions in patients, and the data can be included in the safety analysis;

[0358] (2) Those who do not come to the hospital for reexamination on time or are lost to follow-up, making it impossible to judge the treatment outcome or the test data records are inaccurate, etc., affecting the determination of the treatment effect;

[0359] (3) Those who do not comply with the trial protocol, such as arbitrarily interrupting the treatment protocol or taking other medications that affect the investigational drug;

[0360] (4) Those who are misincluded but do not meet the inclusion criteria.

[0361] 1.7 Random Grouping

[0362] Using SAS software, set the experimental group and the control group in the same proportion. After obtaining the random number table, divide them into the experimental group and the control group according to the random number table method, and include the included cases into the corresponding groups according to the random coding table.

[0363] 1.8 Blinding Design and Emergency Unblinding

[0364] The cases are allocated in a 1:1 ratio between the experimental group and the control group. A two-level blinding method is adopted. The first level is to blind the case serial numbers. Each researcher is assigned a unique case serial number, and the case serial numbers are stored separately from the identity information of the researchers to ensure that only authorized personnel can access them. The second level is that the experimental drugs taken by the two groups have no difference in appearance, and neither the researchers nor the subjects know whether the drug taken is a placebo or a test drug to eliminate the interference of subjective factors. Only the case serial numbers are used to identify the subjects during the implementation of the blinding method, and no information that can identify the identity of the subjects is used. During the blinding process, the written records need to be signed by authorized personnel to maintain the blinded state of the test drugs and placebos and not disclose the true identity of the drugs to the subjects, so as to improve the reliability and credibility of the test results. At the same time, the researchers and the subjects need to sign an informed consent form to clearly know the purpose and method of the test to protect the rights and safety of the subjects.

[0365] This trial is divided into unblinding at the first level before statistical data collection and unblinding at the second level after result statistics. If serious adverse events (SAEs) or serious complications occur during the trial, in order to protect the safety of the subjects and the continuous progress of the trial, emergency unblinding can be carried out. When performing emergency unblinding, it is required that the sponsor monitor, the researcher, the project leader, etc. participate. After unblinding, the time, location, and reason for unblinding should be recorded in detail and signed in a timely manner. This subject will no longer continue to participate in the study, and the test data of this subject cannot be included in the efficacy evaluation, but needs to be included in the safety data analysis. At the same time, timely treatment and protection of the rights of the subject should be provided.

[0366] 1.9 Treatment Plan

[0367] 1.9 Treatment Plan

[0368] Treatment plan for the experimental group: Start taking clomiphene citrate tablets (purchased from the pharmacy of Xiyuan Hospital, China Academy of Chinese Medical Sciences) orally at a dose of 50 mg / time, once a day, for 5 consecutive days on the 5th day of the menstrual cycle; start taking estradiol valerate tablets (purchased from the pharmacy of Xiyuan Hospital, China Academy of Chinese Medical Sciences) orally at a dose of 1 mg / time, once a day, for 10 consecutive days on the 9th day of menstruation. At the same time, take Baoluan Ankun Granules (the granule of the traditional Chinese medicine composition in Example 8) at a dose of 1 bag / time, twice a day, half an hour after meals, and dissolve it with 180 ml of boiling water. Start taking it on the 5th day of the menstrual cycle and take it continuously for 21 days. Monitor the follicles on the 10th day of the menstrual cycle. When the follicle diameter is monitored by ultrasound to be 1.8 cm - 2.0 cm, give 10,000 U of human chorionic gonadotropin by intramuscular injection to induce ovulation. For those without spontaneous menstruation, after excluding pregnancy, monitor the endometrial thickness by vaginal ultrasound. If it is less than 8 mm and there are no follicles larger than 1 cm in both ovaries, start treatment on the same day; if the vaginal ultrasound endometrium is greater than or equal to 8 mm, inject progesterone injection at a dose of 20 mg / day for 3 consecutive days and then stop taking the drug, and start treatment on the 5th day of vaginal bleeding. When the above situation occurs during the treatment period, handle it in the same way.

[0369] Control group ovulation induction protocol: The ovulation induction treatment protocol was the same as that of the treatment group. The traditional Chinese medicine was the simulated Baoluan Ankun Granules (purchased from Beijing Kangrentang Pharmaceutical Co., Ltd., at 5% of the original formula dose). Pregnant patients were considered cured and the treatment was terminated. For non-pregnant patients, the above treatment protocol was continued for 3 menstrual cycles after menstruation. After the treatment was completed, the pregnancy status of the patients was followed up.

[0370] 1.10 Observation indicators

[0371] (1) General clinical data. Observe the basic clinical data of the patients in the experimental group and the control group, including: age, course of disease, menstrual history and marriage and childbearing history.

[0372] (2) Efficacy indicators: pregnancy rate, traditional Chinese medicine syndrome score, AMH and sex hormones, ultrasound indicators, fertility-related score.

[0373] (3) Safety indicators: blood routine, urine routine, liver and kidney function, electrocardiogram.

[0374] 1.11 Treatment of adverse reactions

[0375] Observe whether there are any adverse reactions in the two groups of patients during the observation period. Once any adverse reaction occurs in the patient, the researcher should make timely treatment, analyze the cause of its occurrence, report to the research leader and the ethics committee, and at the same time take corresponding protective measures.

[0376] 1.12 Statistical methods

[0377] SPSS 22.0 software was used for statistical analysis. Measurement data that obeyed the normal or approximately normal distribution were expressed as (x±s) and t-test was used; count data were expressed as rate or constituent ratio and chi-square test was used. Measurement data with skewed distribution and rank data were expressed as median and interquartile range respectively, and rank sum test was used. P<0.05 was considered statistically significant difference.

[0378] 2 Results

[0379] 2.1 Completion of clinical trials

[0380] In this study, a total of 68 cases (34 cases in the experimental group and 34 cases in the control group) were included in the Department of Gynecology, Xiyuan Hospital, China Academy of Chinese Medical Sciences from August 2020 to December 2021. The actual number of completed cases was 65 (34 cases in the experimental group and 31 cases in the control group). The number of dropped cases was 3 (3 cases in the control group), and the dropout rate was 4.61%.

[0381] 2.2 Baseline situation

[0382] 2.2.1 Age and course of disease

[0383] Experimental group: The age distribution was from 26 to 40 years old, with an average age of (34.97 ± 3.42) years. The disease course distribution was from 136 to 110 months, with an average disease course of (33.98 ± 23.98) months.

[0384] Control group: The age distribution was from 26 to 40 years old, with an average age of (35.23 ± 3.88) years. The disease course distribution was from 13 to 92 months, with an average disease course of (36.03 ± 22.62) months.

[0385] There were no statistically significant differences in the age and disease course between the two groups of patients (P > 0.05), and they were comparable.

[0386] 2.2.2 Menstrual history and reproductive history

[0387] Experimental group: Among the subjects, 21 had normal menstruation and 13 had abnormal menstruation (3 cases of premature menstruation, 5 cases of delayed menstruation, and 5 cases of irregular menstruation). In the control group, 22 subjects had normal menstruation and 9 had abnormal menstruation (2 cases of premature menstruation, 6 cases of delayed menstruation, and 1 case of irregular menstruation). Among the experimental group, 14 subjects had primary infertility and 20 had secondary infertility. In the control group, 13 subjects had primary infertility and 18 had secondary infertility. There were no statistically significant differences in the menstrual history and reproductive history between the two groups (P > 0.05), and they were comparable, as shown in Table 40.

[0388] Table 40 Comparison of menstrual history and reproductive history between the two groups

[0389]

[0390] 2.3 Efficacy analysis

[0391] 2.3.1 Pregnancy rate

[0392] In the experimental group, 12 cases were pregnant, 9 cases were pregnant during the treatment period, and 3 cases were pregnant during the follow-up period. The pregnancy rate was 35.29%. In the control group, 4 cases were pregnant, 2 cases were pregnant during the treatment period, and 2 cases were pregnant during the follow-up period. The pregnancy rate was 12.90%. The pregnancy rate of the experimental group was better than that of the control group, and the difference was statistically significant (P > 0.05), as shown in Table 41.

[0393] Table 41 Comparison of pregnancy rates between the two groups

[0394]

[0395] Compared with the control group, *P < 0.05

[0396] 2.3.2 TCM syndrome score

[0397] There was no significant difference in the TCM syndrome scores between the two groups before treatment (P>0.05), and they were comparable. Compared with before treatment in this group, the TCM syndrome scores of both groups decreased after treatment, with significant statistical differences (P<0.01, P<0.01). Compared with the control group, the improvement of symptoms in the experimental group after treatment was significantly better than that in the control group, with a statistical difference (P<0.05). See Table 42.

[0398] Table 42 Comparison of TCM syndrome scores and differences between the two groups

[0399]

[0400] Compared with before treatment, ##P<0.001; compared with the control group, *P<0.05

[0401] 2.2.3 AMH and sex hormones

[0402] During the treatment period, 9 cases in the experimental group were pregnant, 4 cases did not undergo out-of-group examinations after treatment, and 21 patients had data before and after treatment; 2 cases in the control group were pregnant during the treatment period, 3 cases did not undergo out-of-group examinations after treatment, and 26 patients had data before and after treatment.

[0403] There were no obvious differences in AMH and sex hormones between the two groups of patients before treatment (P>0.05), and they were comparable. Compared with before treatment in this group, there were statistically significant differences in AMH levels between the two groups after treatment (P<0.05). Compared with the control group, the increase in AMH in the experimental group was better than that in the control group, with a statistically significant difference (P<0.05). Compared with before treatment in this group, there were no statistically significant differences in FSH and LH levels between the two groups after treatment (P>0.05). Compared with the control group, FSH and LH in the experimental group showed a downward trend, with no statistically significant difference (P>0.05). Compared with before treatment in this group, there were no statistically significant differences in E2 levels between the two groups after treatment (P>0.05). Compared with the control group, E2 in the experimental group increased significantly, with no statistically significant difference (P>0.05). See Table 43.

[0404] Table 43 Comparison of AMH, FSH, LH, E2 and differences between the two groups

[0405]

[0406] Compared with before treatment, #P<0.05; compared with the control group, *P<0.05

[0407] 2.2.4 Comparison of ultrasonic indexes

[0408] During the treatment period, 9 cases in the experimental group were pregnant. Among them, 5 cases did not undergo the exit examination at the end of the treatment, and 20 cases had data before and after treatment. In the control group, 2 cases were pregnant during the treatment period, 6 cases did not undergo the exit examination at the end of the treatment, and 23 cases had data before and after treatment.

[0409] There were no significant differences in the ultrasound indexes between the two groups of patients before treatment (P>0.05), and they were comparable. Compared with before treatment in the same group, the AFC in the experimental group increased by 1.43 after treatment, and the difference was statistically significant (P<0.05). The AFC level in the control group decreased after treatment, and the difference was not statistically significant (P>0.05). Compared with the control group, the improvement degree of AFC in the experimental group was better than that in the control group, and the difference was statistically significant (P<0.05). Compared with before treatment in the same group, there was no significant difference in the improvement of the ovarian stromal blood flow resistance index (RI) between the two groups (P>0.05). Compared with the control group, the RI in the experimental group decreased significantly, but the difference was not statistically significant (P>0.05). See Table 44.

[0410] Table 44 Comparison of AFC, ovarian stromal blood flow resistance index and difference between the two groups

[0411]

[0412] Compared with before treatment, #P<0.05; compared with the control group, *P<0.05

[0413] 2.2.5 Fertility-related scores

[0414] During the treatment period, 9 cases in the experimental group were pregnant. Among them, 7 cases were not continuously monitored until successful ovulation, and 18 cases had continuous ultrasound monitoring of ovulation before and after treatment. In the control group, 2 cases were pregnant during the treatment period, 2 cases were not continuously monitored until successful ovulation, and 11 cases had continuous ultrasound monitoring of ovulation before and after treatment.

[0415] There were no significant differences in the ultrasound indexes between the two groups of patients before treatment (P>0.05), and they were comparable. Compared with before treatment in the same group, there were no significant differences in the ovulation situation score, endometrial morphology score, endometrial thickness score, and endometrial blood flow score between the two groups (P>0.05). Compared with the control group, the ovulation situation score, endometrial thickness score, and endometrial blood flow score in the experimental group showed an increasing trend, but the differences were not statistically significant (P>0.05).

[0416] Compared with before treatment in the same group, the total fertility score in the experimental group increased by 1.61 after treatment, and the difference was statistically significant (P<0.05). The total fertility score in the control group increased by 0.5 after treatment, and the difference was not statistically significant (P>0.05). Compared with the control group, the total fertility score in the experimental group showed an increasing trend, but the difference was not statistically significant (P>0.05). See Table 45 and Table 46.

[0417] Comparison of the scores of endometrial conditions between two groups (Table 45)

[0418]

[0419] Comparison of ovulation conditions and fertility scores between two groups (Table 46)

[0420]

[0421] Compared with before treatment, #P < 0.05

[0422] 2.4 Safety indicators

[0423] There were no obvious abnormalities in the physical and chemical examinations (blood routine, urine routine, liver and kidney function, electrocardiogram) of the experimental group and the control group before and after treatment. No adverse events were observed during the research process.

[0424] 2.5 Follow-up

[0425] After 3 months of follow-up, a total of 59 patients were followed up. Among them, 3 out of 30 patients in the experimental group were pregnant, and 2 out of 29 patients in the control group were pregnant. All pregnant patients in this research were followed up for 1 year: Among the 12 pregnant subjects in the experimental group, 3 were live-born subjects and 9 were continuously pregnant subjects; among the 4 pregnant subjects in the control group, 1 had a fetal arrest, 1 had an ectopic pregnancy, and 2 were continuously pregnant subjects.

[0426] 2.6 Summary

[0427] In summary, the traditional Chinese medicine composition of the present invention can improve the clinical pregnancy rate of patients with infertility related to DOR of liver and kidney yin deficiency, regulate hormone levels, increase AMH and AFC, improve ovarian reserve function, relieve TCM symptoms, and there are no serious adverse reactions before and after treatment, with good safety.

[0428] Obviously, the above-mentioned embodiments of the present invention are merely examples for clearly illustrating the present invention, rather than limitations on the implementation manners of the present invention. For those of ordinary skill in the art, other different forms of changes or modifications can be made based on the above description. It is impossible to list all the implementation manners here. Any obvious changes or modifications derived from the technical solutions of the present invention still fall within the protection scope of the present invention.

Claims

1. A traditional Chinese medicine composition for treating diminished ovarian reserve function, characterized in that, On a parts-by-weight basis, the traditional Chinese medicine composition comprises the following raw medicinal materials: 10-30 parts of stir-fried turtle shell, 10-15 parts of prepared rehmannia root, 6-20 parts of semen cuscutae processed with salt, 9-15 parts of polygonatum sibiricum processed with wine, 10-30 parts of kudzu root, 9-15 parts of polygonum cuspidatum, 6-20 parts of acanthopanax senticosus, 5-15 parts of salvia miltiorrhiza, 6-10 parts of epimedium processed with honey, and 3-10 parts of dried tangerine peel.

2. The traditional Chinese medicine composition according to claim 1, wherein On a parts-by-weight basis, the traditional Chinese medicine composition comprises the following raw medicinal materials: 10 parts of stir-fried turtle shell, 12 parts of prepared rehmannia root, 6 parts of semen cuscutae processed with salt, 9 parts of polygonatum sibiricum processed with wine, 20 parts of kudzu root, 10 parts of polygonum cuspidatum, 6 parts of acanthopanax senticosus, 10 parts of salvia miltiorrhiza, 9 parts of epimedium processed with honey, and 6 parts of dried tangerine peel.

3. The traditional Chinese medicine composition according to claim 1, wherein On a parts-by-weight basis, the traditional Chinese medicine composition comprises the following raw medicinal materials: 15 parts of stir-fried turtle shell, 15 parts of prepared rehmannia root, 10 parts of semen cuscutae processed with salt, 15 parts of polygonatum sibiricum processed with wine, 10 parts of kudzu root, 9 parts of polygonum cuspidatum, 20 parts of acanthopanax senticosus, 5 parts of salvia miltiorrhiza, 6 parts of epimedium processed with honey, and 9 parts of dried tangerine peel.

4. The traditional Chinese medicine composition according to claim 1, characterized in that, On a parts-by-weight basis, the traditional Chinese medicine composition comprises the following raw medicinal materials: 30 parts of stir-fried turtle shell, 10 parts of prepared rehmannia root, 20 parts of semen cuscutae processed with salt, 10 parts of polygonatum sibiricum processed with wine, 30 parts of kudzu root, 12 parts of polygonum cuspidatum, 15 parts of acanthopanax senticosus, 15 parts of salvia miltiorrhiza, 10 parts of epimedium processed with honey, and 10 parts of dried tangerine peel.

5. The traditional Chinese medicine composition according to claim 1, wherein On a parts-by-weight basis, the traditional Chinese medicine composition comprises the following raw medicinal materials: 30 parts of stir-fried turtle shell, 15 parts of prepared rehmannia root, 20 parts of semen cuscutae processed with salt, 10 parts of polygonatum sibiricum processed with wine, 30 parts of kudzu root, 10 parts of polygonum cuspidatum, 10 parts of acanthopanax senticosus, 15 parts of salvia miltiorrhiza, 10 parts of epimedium processed with honey, and 10 parts of dried tangerine peel.

6. A pharmaceutical preparation, characterized in that, The pharmaceutical preparation contains the traditional Chinese medicine composition according to any one of claims 1-5.

7. The pharmaceutical preparation according to claim 6, characterized in that, The pharmaceutical preparation contains a pharmaceutically acceptable carrier or excipient.

8. The pharmaceutical preparation according to claim 6 or 7, characterized in that, The dosage form of the pharmaceutical preparation is a decoction, a medicinal broth, granules, pills, capsules, tablets, powders or oral liquids.

9. Use of a traditional Chinese medicine composition according to any one of claims 1-5 in the preparation of a drug for treating ovarian reserve function decline and related diseases.

10. The use according to claim 9, characterized in that, The ovarian reserve function decline-related diseases include infertility; the ovarian reserve function decline is of the liver-kidney yin deficiency type.

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