Application of a traditional Chinese medicine composition in the preparation of a drug for improving pregnancy outcomes in patients with thin endometrium after frozen-thawed embryo transfer.

Abstract

This invention relates to the field of traditional Chinese medicine technology, and more particularly to the application of a traditional Chinese medicine composition in the preparation of a drug for improving pregnancy outcomes in frozen-thawed embryo transfer in patients with thin endometrium. Randomized controlled clinical trials have shown that the traditional Chinese medicine composition of this invention can significantly improve symptoms of kidney deficiency and blood stasis, promote endometrial growth and development in patients with thin endometrium, improve endometrial blood perfusion, and improve pregnancy outcomes by enhancing endometrial receptivity, with good safety profile.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine technology, and in particular to the application of a traditional Chinese medicine composition in the preparation of a drug for improving pregnancy outcomes in patients with thin endometrium undergoing frozen-thawed embryo transfer. Background Technology

[0002] Thin endometrium is a significant factor contributing to embryo implantation failure and declining pregnancy rates with assisted reproductive technologies. It is generally believed that insufficient endometrial thickness, poor blood perfusion, and decreased receptivity during embryo transfer cycles can all lead to implantation failure. Current clinical treatments primarily employ estrogen supplementation, methods to improve endometrial blood flow, and local intrauterine therapy, but their effectiveness is limited, and some patients still struggle to achieve satisfactory pregnancy outcomes. Therefore, developing safe and effective treatment options that improve endometrial receptivity is of great importance.

[0003] Based on the fundamental treatment principles of "tonifying the kidneys" and "invigorating blood circulation," the applicant's herbal composition (prepared licorice root, vinegar-processed five-spice powder, prepared ginger, chicken blood vine, salvia miltiorrhiza, wine-processed cornus officinalis, bran-fried yam, wine-processed cistanche deserticola, salt-processed psoralea corylifolia, deer antler powder, codonopsis pilosula, and euryale ferox) consists of 12 Chinese herbs. Its purpose is to tonify the kidneys, invigorate blood circulation, replenish essence and nourish the uterus, increase the thickness of the endometrium in women, and improve their ability to conceive. Randomized controlled clinical trials have shown that it can improve endometrial receptivity and increase pregnancy outcomes. Summary of the Invention

[0004] This invention provides the application of a traditional Chinese medicine composition in the preparation of a drug for improving pregnancy outcomes in frozen-thawed embryo transfer in patients with thin endometrium, thus solving the problems existing in the prior art.

[0005] The technical solution adopted in this invention is:

[0006] The application of a traditional Chinese medicine composition in the preparation of a drug for improving pregnancy outcomes in patients with thin endometrium undergoing frozen-thawed embryo transfer, wherein the traditional Chinese medicine composition improves pregnancy outcomes by enhancing endometrial receptivity; the traditional Chinese medicine composition is made from the following raw materials in parts by weight: 10-20 parts of deer antler powder, 10-20 parts of Codonopsis pilosula, 10-20 parts of salt-processed Psoralea corylifolia, 10-20 parts of wine-processed Cornus officinalis, 8-15 parts of wine-processed Cistanche deserticola, 3-10 parts of processed ginger, 20-30 parts of bran-fried Dioscorea opposita, 10-25 parts of Spatholobus suberectus, 3-10 parts of processed licorice root, 10-25 parts of Salvia miltiorrhiza, 10-20 parts of vinegar-processed Trogopterus xanthipes, and 10-20 parts of Euryale ferox.

[0007] Furthermore, the traditional Chinese medicine composition is made from the following raw materials in parts by weight: 15 parts deer antler powder, 15 parts codonopsis, 15 parts salt-processed psoralea, 15 parts wine-processed cornus officinalis, 12 parts wine-processed cistanche deserticola, 6 parts prepared ginger, 24 parts bran-fried yam, 18 parts chicken blood vine, 6 parts prepared licorice root, 18 parts salvia miltiorrhiza, 15 parts vinegar-processed five-spice powder, and 15 parts euryale ferox.

[0008] Furthermore, the dosage form of the traditional Chinese medicine composition is granules, decoction, powder, capsule, tablet or pill; granules are preferred.

[0009] Furthermore, the traditional Chinese medicine composition improves embryo implantation by activating the PPAR pathway, increasing the expression of PPARα and PPARγ, thereby improving or treating pregnancy outcomes in patients with thin endometrium undergoing frozen-thawed embryo transfer.

[0010] Furthermore, the above-mentioned application of the traditional Chinese medicine composition has achieved significant therapeutic effects in patients with thin endometrium, increasing endometrial thickness and volume, improving endometrial blood flow perfusion parameters VI, FI and VFI, improving uterine artery blood flow PI, RI and S / D, increasing embryo implantation rate, and improving clinical pregnancy rate and continuous pregnancy rate.

[0011] Furthermore, the application is the use of the traditional Chinese medicine composition in combination with estrogen therapy in the preparation of a drug for improving pregnancy outcomes in frozen-thawed embryo transfer in patients with thin endometrium.

[0012] The beneficial effects of this invention are:

[0013] This invention is formulated based on the principle of "tonifying the kidneys and promoting blood circulation, replenishing essence and nourishing the uterus." It consists of deer antler powder, codonopsis, salt-processed psoralea, wine-processed cornus, wine-processed cistanche, prepared ginger, bran-fried yam, chicken blood vine, prepared licorice root, salvia miltiorrhiza, vinegar-processed five-spice powder, and euryale seed. It is designed for patients with thin endometrium and kidney deficiency with blood stasis. The formula is based on "Yulinzhu" (ginseng, atractylodes macrocephala, poria cocos, white peony root, chuanxiong rhizome, prepared licorice root, angelica sinensis, prepared rehmannia root, cuscuta chinensis, eucommia ulmoides, deer antler powder, and Sichuan pepper), taking its core principles of tonifying the kidneys and spleen, and regulating the Chong and Ren meridians. Because patients may experience stomach upset or significant blood stasis, angelica sinensis and prepared rehmannia root, which are relatively nourishing, are temporarily removed to prevent stagnation. Since yam and codonopsis have already been used to strengthen the spleen and replenish qi, poria cocos and atractylodes macrocephala are removed to avoid dispersing the medicinal effects. The addition of Psoralea corylifolia and Cistanche deserticola significantly enhances the kidney-warming and yang-tonifying effects. The combination of Salvia miltiorrhiza and Spatholobus suberectus in the "Shaofu Zhuyu Tang" formula replaces the original Ligusticum chuanxiong and Angelica sinensis, offering a more balanced and potent approach to blood-activating, blood-nourishing, and meridian-clearing effects. Salvia miltiorrhiza nourishes and activates blood, while Spatholobus suberectus clears the meridians and nourishes blood, forming a "combination of attack and tonification" blood-activating structure, expanding its applicability to chronic diseases with blood deficiency and blood stasis. The addition of Trogopterus xanthipes enhances the effects of resolving blood stasis and relieving pain, directly targeting the affected area. Optimization was also made for the selection of herbs with different processing methods: salt-processed herbs guide the medicine to the kidneys, with salt-processed Psoralea corylifolia chosen to specifically target the lower burner; wine-processed Cistanche deserticola and Cornus officinalis are chosen because wine processing enhances their warming, unblocking, and blood-activating effects; stir-fried Dioscorea opposita with wheat bran further emphasizes its spleen-strengthening and qi-tonifying effects; vinegar-processed Trogopterus xanthipes enhances its blood-stasis-resolving and pain-relieving effects while reducing its fishy and dry odor, hence the choice of vinegar-processed Trogopterus xanthipes. Drug dosage adjustment: Increase the dosage of yam (24g), salvia miltiorrhiza (18g), and chicken blood vine (18g) to highlight the key role of strengthening the spleen and promoting blood circulation in this formula, and ensure the dosage. The final result is a compound formula structure with kidney tonification and warming yang as the foundation, blood circulation and collaterals promotion as the function, and simultaneous treatment of the spleen and kidney as the adjuvant.

[0014] The traditional Chinese medicine composition of this invention has been shown to have the following therapeutic effects through randomized controlled clinical trials:

[0015] (1) Significantly improves symptoms of kidney deficiency and blood stasis

[0016] After treatment, the TCM syndrome scores of both the experimental and control groups decreased compared to before treatment, but the decrease in the experimental group was significantly greater than that in the control group. The total effective rate of the experimental group reached 69.70%, which was significantly higher than that of the control group (32.43%), indicating that the TCM composition of this invention can significantly improve symptoms related to kidney deficiency and blood stasis, such as soreness and weakness of the lower back and knees, dizziness and tinnitus, and cold limbs.

[0017] (2) Promotes endometrial growth and development

[0018] After treatment, the endometrial thickness and volume in the experimental group significantly increased compared to before treatment, and the increase was greater than that in the control group. The results indicate that the herbal composition of this invention can promote endometrial growth and improve the underdeveloped endometrial condition in patients with thin endometrium.

[0019] (3) Improve endometrial blood perfusion

[0020] After treatment, the endometrial blood flow grading in the experimental group was significantly improved, and the endometrial blood flow parameters VI, FI, and VFI all showed an increasing trend and were higher than those in the control group. The results suggest that the traditional Chinese medicine composition of this invention can improve local microcirculation and blood perfusion in the endometrium, and enhance endometrial receptivity.

[0021] (4) Improve assisted reproductive outcomes

[0022] After treatment, the embryo implantation rate, clinical pregnancy rate, and sustained pregnancy rate in the experimental group were all higher than those in the control group, indicating that the traditional Chinese medicine composition of the present invention is beneficial to improving the pregnancy outcome of frozen-thawed embryo transfer cycles in patients with thin endometrium.

[0023] (5) Good safety

[0024] No serious adverse reactions were found during the clinical study, and the patients tolerated it well, indicating good prospects for clinical application.

[0025] In summary, the traditional Chinese medicine composition granules provided by this invention can improve the endometrial receptivity of patients with thin endometrium by improving symptoms of kidney deficiency and blood stasis, promoting endometrial growth, and improving endometrial blood perfusion, thereby improving the outcome of assisted reproductive treatment. Attached Figure Description

[0026] Figure 1 This invention compares the uterine morphology and pregnancy status of rats in different groups.

[0027] Figure 2The pregnancy rate and embryo implantation status of rats in each group of this invention;

[0028] Figure 3 HE staining of the endometrium of rats in each group of this invention;

[0029] Figure 4 This invention provides statistics on endometrial thickness and glandular number in each group of rats.

[0030] Figure 5 This invention describes the endometrial microvascular formation in rats from each group.

[0031] Figure 6 The expression of E-cadherin, LIF and Vimentin in the endometrium of rats in each group of this invention;

[0032] Figure 7 The correlation between endometrial volume and endometrial thickness after treatment according to the present invention;

[0033] Figure 8 This invention relates to the correlation between endometrial VI and endometrial thickness after treatment.

[0034] Figure 9 This invention relates to the correlation between endometrial fibrillation (FI) and endometrial thickness after treatment.

[0035] Figure 10 This invention relates to the correlation between endometrial VFI and endometrial thickness after treatment.

[0036] in,

[0037] Figure 1 The two rows of images in the middle, from left to right, correspond to the blank group (Control), the model group (Model), the low-dose group of the traditional Chinese medicine composition of the present invention (PBYST-L), the medium-dose group of the traditional Chinese medicine composition of the present invention (PBYST-M), the high-dose group of the traditional Chinese medicine composition of the present invention (PBYST-H), and the aspirin group;

[0038] Figure 2 In the middle section, A represents pregnancy rate statistics; B represents embryo implantation site statistics.

[0039] Figure 4 In the middle section, A represents the statistics on endometrial thickness; B represents the statistics on the number of endometrial glands.

[0040] Figure 5 In the table, A represents the expression of CD34 and VEGFA in the endometrium of rats in each group detected by IHC; B represents the CD34 staining MVD count; and C represents the VEGFA staining histochemical score.

[0041] Figure 6In the table, A represents the expression of E-cadherin, LIF, and Vimentin in the endometrium of rats in each group detected by IHC; B represents the statistical analysis of E-cadherin staining histochemical scores; C represents the statistical analysis of LIF staining histochemical scores; and D represents the statistical analysis of Vimentin staining histochemical scores. Detailed Implementation

[0042] To more clearly illustrate the overall concept of the present invention, a detailed description is provided below with reference to the accompanying drawings. The embodiments described below are merely preferred embodiments of the present invention. It should be noted that the following description is solely for explaining the present invention and does not constitute any limitation on the present invention. Any simple modifications, equivalent variations, and alterations made to the embodiments based on the technical essence of the present invention fall within the scope of the present invention.

[0043] Unless otherwise specified, the instruments, reagents, and materials used in the following embodiments are all conventional instruments, reagents, and materials already available in the prior art and can be obtained through legitimate commercial channels. Unless otherwise specified, the experimental methods and detection methods used in the following embodiments are all conventional experimental methods and detection methods already available in the prior art.

[0044] Example 1

[0045] This traditional Chinese medicine composition granule, named PBYST, is made from the following raw materials by weight: 15g of deer antler powder, 15g of codonopsis, 15g of salt-processed psoralea, 15g of wine-processed cornus officinalis, 12g of wine-processed cistanche deserticola, 6g of prepared ginger, 24g of bran-fried yam, 18g of chicken blood vine, 6g of prepared licorice root, 18g of salvia miltiorrhiza, 15g of vinegar-processed five-spice powder, and 15g of euryale ferox.

[0046] The above methods for preparing granules are as follows:

[0047] Take all the above-mentioned medicinal materials, put them into an extraction tank, add 10 times the amount of purified water and soak at room temperature for 60 minutes; for the first decoction, heat to boiling and then maintain a gentle boil for 60 minutes, filter out the first decoction; add 8 times the amount of purified water to the residue, boil for the second time and then maintain a gentle boil for 40 minutes, filter out the second decoction; combine the two decoctions, filter through a 120-mesh filter to remove residue, concentrate the filtrate under reduced pressure and low temperature (vacuum degree -0.06~-0.08MPa, temperature 60~70℃) to obtain an extract with a relative density of 1.18~1.22 (measured at 60℃); add appropriate amounts of dextrin and sucrose as excipients to the extract, stir thoroughly to make a soft mass; use an extrusion granulator to pass through a 14-mesh sieve to make wet granules; place the wet granules in a hot air circulating oven and dry at a low temperature of 60℃ until the moisture content is ≤5.0%; after drying, use 12-mesh and 24-mesh sieves to granulate, remove fine powder and large lumps; mix all the obtained granules evenly and divide them into 2 bags for independent packaging.

[0048] Dosage and administration: Take one packet twice a day, dissolved in warm water 30 minutes after breakfast and dinner.

[0049] Example 2

[0050] This traditional Chinese medicine composition in granule form, named PBYST, is made from the following raw materials by weight:

[0051] 20g of deer antler powder, 20g of codonopsis, 20g of salt-processed psoralea, 20g of wine-processed cornus officinalis, 15g of wine-processed cistanche deserticola, 10g of prepared ginger, 30g of wheat bran-fried yam, 25g of chicken blood vine, 10g of prepared licorice root, 25g of salvia miltiorrhiza, 20g of vinegar-processed five-spice powder, and 20g of euryale ferox.

[0052] For preparation methods and dosage, please refer to Implementation 1. This formula has a strong effect in warming the kidneys and promoting blood circulation, and is suitable for patients with severe symptoms of kidney deficiency and blood stasis.

[0053] Example 3

[0054] This traditional Chinese medicine composition in granule form, named PBYST, is made from the following raw materials by weight:

[0055] 10g of deer antler powder, 10g of codonopsis, 10g of salt-processed psoralea, 10g of wine-processed cornus officinalis, 8g of wine-processed cistanche deserticola, 3g of prepared ginger, 20g of wheat bran-fried yam, 10g of chicken blood vine, 3g of prepared licorice root, 10g of salvia miltiorrhiza, 10g of vinegar-processed five-spice powder, and 10g of euryale ferox.

[0056] For preparation methods and dosage, please refer to Implementation 1. This formula has a mild and gentle nature and is suitable for patients with mild symptoms of kidney deficiency and blood stasis.

[0057] The following experimental study and verification were conducted using animal experiments and clinical observations. The traditional Chinese medicine composition of Example 1 of the present invention was selected to study and verify the effect of the traditional Chinese medicine composition of the present invention and the sample prepared by the reference drug on improving the pregnancy outcome of frozen-thawed embryo transfer in patients with thin endometrium.

[0058] I. Animal Experiments

[0059] 1.1 Experimental Objective

[0060] A rat embryo implantation disorder model was constructed, and the effect of the traditional Chinese medicine composition granules of Example 1 of this invention on the endometrial receptivity of rats with embryo implantation disorder was investigated through experiments. The efficacy and basic principle of the traditional Chinese medicine composition in improving embryo implantation disorder were evaluated.

[0061] 1.2 Laboratory Animals

[0062] Forty-eight SPF-grade 7-week-old SD female rats weighing 220±20 g and twenty-four male rats of the same age weighing 300±20 g with reproductive ability were provided by Jinan Pengyue Laboratory Animal Breeding Co., Ltd., with the license number SCXK (Lu) 20220006. All rats were housed in the SPF animal barrier system of the Animal Center of Affiliated Hospital of Shandong University of Traditional Chinese Medicine. All rats were adaptively fed for 1 week and had free access to food and water. All animal experiments were approved by the Ethics Committee of Affiliated Hospital of Shandong University of Traditional Chinese Medicine.

[0063] 1.3. Model establishment

[0064] On the morning of the 4th day of pregnancy (equivalent to the clinical embryo implantation window period), the rats in the model group, each dose group of traditional Chinese medicine, and the aspirin group were subcutaneously injected with a 2 mg / ml mifepristone oil solution (5.5 mg / kg) in the neck to create an embryo implantation obstacle model, and the rats in the blank group were given the same dose of sesame oil at the same time.

[0065] 1.4. Experimental grouping

[0066] After 1 week of adaptive feeding, vaginal smears of female rats were observed for 1 week to observe the estrous cycle of rats. Rats with regular estrous cycles were selected. At 5:00 pm every day, male and female rats were caged at a ratio of 2:1. At 8:00 am the next day, vaginal plugs were observed and vaginal smears were taken to check for sperm. If a vaginal plug or sperm was found, it was recorded as the first day of mating. After 5 days of observation, female rats that did not mate were excluded. The mated female rats were randomly divided into 6 groups: blank group (Control), model group (Model), low-dose group of the traditional Chinese medicine composition of the present invention (PBYST-L), medium-dose group of the traditional Chinese medicine composition of the present invention (PBYST-M), high-dose group of the traditional Chinese medicine composition of the present invention (PBYST-H), and aspirin group (ASPL), with 8 rats in each group.

[0067] 1.5. Administration method

[0068] All female rats with regular estrous cycles were selected for 2:1 male-female mating, and the mating time was recorded. From the first day of mating, continuous gavage was performed until the sixth day of mating.

[0069] The dosage of traditional Chinese medicine was calculated based on an adult of 60 kg, with a daily dosage of 80 g. The equivalent dose for rats was 6.3 times the dosage per kilogram of body weight for clinical adults, that is, 0.84 g / (100 g·day). The high-dose group of the traditional Chinese medicine composition of the present invention was given gavage at a dose of 1.68 g / (100 g·day), the medium-dose group of the traditional Chinese medicine composition of the present invention was given gavage at a dose of 0.84 g / (100 g·day), and the low-dose group of the traditional Chinese medicine composition of the present invention was given gavage at a dose of 0.42 g / (100 g·day).

[0070] Rats in the aspirin group were administered 0.79 mg / (100 g·day) by gavage (equivalent to 75 mg / 60 kg in adults).

[0071] The model group was given 0.8 ml / (100g·day) of distilled water by gavage according to their body weight.

[0072] 1.6. Observation Indicators

[0073] 1.6.1 Pregnancy rate and embryo implantation status

[0074] 1.6.2 Endometrial thickness and number of glands

[0075] 1.6.3 Endometrial angiogenesis

[0076] 1.6.4 Expression of endometrial receptivity markers

[0077] 1.7. Experimental Results

[0078] 1.7.1 Pregnancy rate and embryo implantation status in each group of rats

[0079] In the control group, the number of embryos implanted in the uterus of rats was relatively high, and the uterine body at the implantation site was enlarged, resembling a string of beads. In the model group, the number of embryos implanted in the uterus was extremely low, and the uterus of rats in the dosage groups of the herbal composition of this invention showed a small number of enlarged areas. In the medium and high dosage groups of the herbal composition of this invention and the aspirin group, the uterine body of rats was relatively long, and there were many embryo implantation sites.

[0080] See Figure 1 To assess the uterine morphology and pregnancy status of rats in each group, the pregnancy rate and number of embryonic implantation sites were statistically analyzed. The results are shown in Table 1 below. The highest pregnancy rate was 87.5% in the control group and the lowest was 37.5% in the model group. The average number of embryonic implantation sites was highest in the control group (11.00±3.56) and lowest in the model group (2.67±1.53), with a statistically significant difference (P<0.05). The number of embryonic implantation sites in the medium-dose group and the aspirin group of the herbal composition of this invention was significantly increased compared to the model group, with statistically significant differences (P<0.05). Figure 2 Statistical charts showing pregnancy rate and embryo implantation status in each group of rats.

[0081] Table 1 Comparison of pregnancy rate and number of embryo implantation sites in rats of different groups ( ±s)

[0082]

[0083] 1.7.2 Endometrial thickness and glandular number in each group of rats

[0084] The results of HE staining of the endometrium of rats in each group were observed under a light microscope as follows: Figure 3As shown: The endometrial, myometrial, and serosa layers of the rats in each group were basically intact. Measurements of endometrial thickness revealed no significant difference among the groups (e.g., ...). Figure 4 (A). Statistical analysis of the number of glands revealed that the number of glands in the model group was significantly lower than that in the blank group, the medium-dose group of the herbal composition of this invention, and the aspirin group, with statistically significant differences (P<0.05). Figure 4 (B) This indicates that mifepristone modeling does not damage endometrial thickness, but may damage endometrial glands. The herbal composition of this invention can promote gland formation.

[0085] 1.7.3 Endometrial angiogenesis

[0086] Endometrial angiogenesis in rats of different groups was assessed by immunohistochemical staining for CD34 and VEGFA. CD34 expression is commonly used to assess microvessel density. Using a digital section scanner, all tissue information on CD34-immunohistochemically treated sections was scanned. Target areas were selected for 200x magnification imaging, and microvessels were counted in three fields of view for each section. The average MVD value (vessels / HP) was calculated. See also... Figure 5 In the study, the results showed that the MVD in the model group was significantly lower than that in the control group (P<0.05). VEGFA is a vascular endothelial growth factor, such as... Figure 5 The results showed that the immunohistochemical score of the model group was significantly lower than that of the blank group (P<0.01), and the immunohistochemical scores of the medium-dose group and the high-dose group of the traditional Chinese medicine composition of the present invention were higher than those of the model group (P<0.05).

[0087] 1.7.4 Expression of endometrial receptivity markers E-cadherin, LIF, and Vimentin

[0088] The endometrial receptivity of rats in each group was assessed by immunohistochemical staining for E-cadherin, LIF, and Vimentin. Figure 6 The immunohistochemical score of the A. E-cadherin staining model group was lower than that of the blank group, and the difference was statistically significant (P<0.01). Figure 6 As shown in Figure B, the immunohistochemical score of the LIF staining model group was lower than that of the blank group, and the difference was statistically significant (P<0.01). Figure 6 As shown in Figure C, the immunohistochemical scores of the dosage group and the aspirin group of the traditional Chinese medicine composition of this invention were higher than those of the model group, and the differences were statistically significant (P<0.05). The immunohistochemical score of the Vimentin staining model group was lower than that of the blank group, and the difference was statistically significant (P<0.01). Figure 6The results showed that after using the medium dose of the traditional Chinese medicine composition of the present invention, the immunohistochemical score was higher than that of the model group, and the difference was statistically significant (P<0.05).

[0089] 1.8. Experimental Conclusions

[0090] The dosage of the traditional Chinese medicine composition of this invention can increase the number of endometrial glands in rats with embryo implantation disorders, promote angiogenesis, increase the expression of LIF and Vimentin in the rat endometrium, and improve embryo implantation.

[0091] II. Clinical Research Trials

[0092] 2.1. Materials and Methods

[0093] This study included 131 patients with thin endometrial tissue and infertility who visited the Integrated Traditional Chinese and Western Medicine Reproductive and Genetic Center of the Affiliated Hospital of Shandong University of Traditional Chinese Medicine between January 2019 and December 2024. These patients were diagnosed with kidney deficiency and blood stasis syndrome according to Traditional Chinese Medicine (TCM) diagnosis. They were divided into an experimental group (treated with the TCM composition of this invention combined with estrogen therapy) and a control group (treated with estrogen alone), with 71 patients in the experimental group and 60 patients in the control group. Inclusion criteria: ① Meeting the diagnostic criteria for infertility, thin endometrium, and kidney deficiency and blood stasis syndrome according to TCM diagnosis; ② Age 25-35 years, with fertility requirements; ③ Patients undergoing frozen-thawed embryo transfer; ④ Number of embryos transferred per cycle: 1-2 grade I or II embryos; ⑤ No use of hormonal drugs within the past 3 months.

[0094] 2.2. Statistical Methods

[0095] SPSS 26.0 software was used for statistical analysis. For normally distributed count data, t-tests were used for comparison; for non-normally distributed data, nonparametric tests were used. All data are expressed as mean ± standard deviation (x ± s). Count data were expressed as frequencies. Fisher's exact test was used for data with a total sample size less than 40 or a minimum expected frequency less than 5; the chi-square test was used for data with a total sample size greater than 40 and a minimum expected frequency greater than 5. A p-value < 0.05 was considered statistically significant; a p-value < 0.01 was considered highly statistically significant.

[0096] 2.3. Results

[0097] 2.3.1 Comparison of baseline data between the experimental group and the control group

[0098] There were no statistically significant differences between the experimental and control groups on endometrial thickness, history of dilation and curettage (D&C), age, body mass index (BMI), duration of infertility, basal follicle-stimulating hormone (bFSH), basal luteinizing hormone (bLH), basal estradiol (bE2), and basal progesterone (bP) on the inclusion date (P > 0.05). See Table 2.

[0099] Table 2 Basic Data Analysis Table ±s)

[0100]

[0101] 2.3.2 There were no statistically significant differences between the experimental group and the control group in the number of D3 and D5 embryos transferred, or in the total number of grade I and grade II embryos transferred (P > 0.05). See Table 3.

[0102] Table 3 Comparison of the number of embryos transferred

[0103]

[0104] 2.3.3 There was no statistically significant difference in the scores of kidney deficiency and blood stasis syndrome between the experimental group and the control group before treatment (P>0.05). See Table 4.

[0105] Table 4 Comparison of syndrome scores before treatment ( ±s)

[0106]

[0107] 2.3.4 Comparison of ultrasound indicators between the experimental group and the control group before treatment: endometrial thickness and volume, endometrial VI, FI, VFI, subendometrial blood flow PI, RI, S / D, and uterine artery blood flow PI, RI, S / D. There were no statistically significant differences between the two groups (P>0.05), and they were comparable, as shown in Table 5.

[0108] Table 5 Comparison of ultrasound indicators before treatment ( ±s)

[0109]

[0110] 2.3.5 There were no statistically significant differences in endometrial blood flow grading and peristalsis between the experimental group and the control group before treatment (P>0.05). See Tables 6 and 7.

[0111] Table 6 Comparison of intimal blood flow grading before treatment

[0112]

[0113] Table 7 Comparison of endometrial peristalsis before treatment

[0114]

[0115] 2.3.6 Evaluation of Traditional Chinese Medicine Syndromes

[0116] There were highly significant differences in the syndrome scores of kidney deficiency and blood stasis before and after treatment in both groups (P < 0.01); at the same time, the syndrome score of kidney deficiency and blood stasis in the experimental group was significantly lower than that in the control group after treatment (16.70 ± 3.20 vs. 19.19 ± 2.21) (P < 0.01). See Table 8.

[0117] Table 8 Comparison of syndrome scores before and after treatment ( ±s)

[0118]

[0119] Note: **The difference is statistically significant (P < 0.01)**

[0120] The chi-square test showed a highly significant difference in efficacy between the two groups (P=0.004<0.01), with the effective rate in the experimental group being significantly higher than that in the control group.

[0121] 2.3.7 Comparison of endometrial thickness

[0122] Comparing the two groups before and after treatment, there were highly significant differences in endometrial thickness (P < 0.01); there was no statistically significant difference in endometrial thickness between the groups after treatment (P > 0.05), but the increase in endometrial thickness in the experimental group was more significant than that in the control group (0.83 ± 0.11 vs. 0.82 ± 0.09). See Table 9.

[0123] Table 9 Comparison of endometrial thickness (cm) before and after treatment ±s)

[0124]

[0125] Note: **The difference is statistically significant (P < 0.01)**

[0126] 2.3.8 Comparison of endometrial volume

[0127] Comparing the two groups before and after treatment, there were highly significant differences in endometrial volume (P < 0.01); however, there was no statistically significant difference in endometrial volume after treatment between the groups (P > 0.05), but the experimental group showed a more significant increase in EMV compared to the control group (2.53 ± 0.77 vs. 2.44 ± 0.83). See Table 10. Post-treatment EMV was linearly correlated with endometrial thickness (R² = 0.676). Figure 7 .

[0128] Table 10 Comparison of endometrial volume (cm3) before and after treatment ±s)

[0129]

[0130] Note: **The difference is statistically significant (P < 0.01)**

[0131] 2.3.9 Comparison of endometrial blood flow

[0132] There was a statistically significant difference in endothelial blood flow grading before and after treatment in the experimental group (P < 0.05), while there was no statistically significant difference in endothelial blood flow grading before and after treatment in the control group (P > 0.05). There was also no statistically significant difference in endothelial blood flow grading after treatment between the two groups (P > 0.05). See Table 11.

[0133] Table 11 Comparison of intimal blood flow grading before and after treatment

[0134]

[0135] Note: *Statistically significant difference (P < 0.05)

[0136] 2.3.10 Comparison of endometrial peristalsis

[0137] There were no statistically significant differences in endometrial peristalsis before and after treatment in either group (P>0.05), and there were also no statistically significant differences in endometrial peristalsis after treatment in either group (P>0.05). The number of patients with quiescent endometrial peristalsis in the experimental group increased after treatment compared to before treatment (20 / 33 vs. 14 / 33), while the number of patients with quiescent endometrial peristalsis in the control group remained unchanged after treatment compared to before treatment (18 / 37 vs. 18 / 37). See Table 12.

[0138] Table 12 Comparison of endometrial peristalsis before and after treatment

[0139]

[0140] 2.3.11 Comparison of other three-dimensional ultrasound related indicators

[0141] In the experimental group, there were highly statistically significant differences in endometrial VI, FI, VFI, and subendometrial blood flow PI before and after treatment (P < 0.01), while other three-dimensional ultrasound indicators showed no statistically significant differences. See Table 13.

[0142] Table 13 Comparison of ultrasound indicators before and after treatment in the experimental group ( ±s)

[0143]

[0144] Note: **The difference is statistically significant (P < 0.01)**

[0145] In the control group, there were highly statistically significant differences in endometrial VI, FI, and VFI before and after treatment (P < 0.01). See Table 14.

[0146] Table 14 Comparison of ultrasound indicators before and after treatment in the control group ( ±s)

[0147]

[0148] Note: **The difference is statistically significant (P < 0.01)**

[0149] There were no statistically significant differences in any of the ultrasound indicators after treatment between the groups (P > 0.05). See Table 15.

[0150] Table 15 Comparison of ultrasound indicators after treatment ( ±s)

[0151]

[0152] 3.12 Correlation between endometrial blood flow and endometrial thickness after treatment

[0153] After treatment, endometrial VI (R²=0.662), FI (R²=0.519), and VFI (R²=0.749) were all linearly correlated with endometrial thickness. See also... Figure 8 , Figure 9 , Figure 10 .

[0154] 2.3.13 Comparison of Clinical Efficacy Indicators

[0155] There was no significant difference in clinical outcomes between the two groups after treatment (P>0.05), but the embryo implantation rate, clinical pregnancy rate, and sustained pregnancy rate were all higher in the experimental group than in the control group. See Table 16.

[0156] Table 16 Comparison of clinical indicators between the two groups of patients

[0157]

[0158] 2.3.14 Safety Evaluation

[0159] Three patients in all enrolled patients participated in the study and experienced gastrointestinal adverse reactions during the observation period: nausea, vomiting, abdominal distension, and diarrhea. These symptoms disappeared without intervention after discontinuation of the medication. No grade 3 or higher adverse reactions occurred in either group, indicating that the herbal composition granules of this invention have a high safety profile for improving endometrial thickness in patients with thin endometrium due to kidney deficiency and blood stasis.

[0160] 2.4. Summary

[0161] This retrospective study demonstrated the effectiveness of the herbal composition of this invention in patients with thin endometrium due to kidney deficiency and blood stasis. Regarding the efficacy of the herbal medicine, the experimental group showed significantly lower scores for kidney deficiency and blood stasis syndrome compared to the control group after treatment, and the clinical effective rate was significantly higher in the experimental group, proving that the herbal medicine can significantly improve the kidney deficiency and blood stasis syndrome in patients. In terms of improving FET (fetal implantation) outcomes, the embryo implantation rate and clinical pregnancy rate in the experimental group were higher than those in the control group. 3D-US examinations were performed on all patients before and after treatment. The results showed that the endometrial blood flow grading and subendometrial blood flow PI were significantly improved in the experimental group compared to the control group after treatment. The herbal medicine also had a certain improving effect on endometrial thickness and volume, endometrial blood flow VI, FI, and VFI. The combination of herbal medicine and estrogen therapy can promote endometrial growth in patients with thin endometrium in multiple ways, thereby improving endometrial receptivity and increasing the embryo implantation rate and clinical pregnancy rate.

[0162] The above specific embodiments should not be construed as limiting the scope of protection of the present invention. For those skilled in the art, any alternative improvements or modifications made to the embodiments of the present invention shall fall within the scope of protection of the present invention.

[0163] Any aspects of this invention not described in detail are well-known to those skilled in the art.

Claims

1. The application of a traditional Chinese medicine composition in the preparation of a drug for improving pregnancy outcomes in frozen-thawed embryo transfer in patients with thin endometrium, characterized in that, The traditional Chinese medicine composition improves pregnancy outcomes by enhancing endometrial receptivity; the traditional Chinese medicine composition is made from the following raw materials in parts by weight: 10-20 parts of deer antler powder, 10-20 parts of codonopsis pilosula, 10-20 parts of salt-processed psoralea corylifolia, 10-20 parts of wine-processed cornus officinalis, 8-15 parts of wine-processed cistanche deserticola, 3-10 parts of processed ginger, 20-30 parts of bran-fried yam, 10-25 parts of chicken blood vine, 3-10 parts of processed licorice root, 10-25 parts of salvia miltiorrhiza, 10-20 parts of vinegar-processed trogopterus xanthipes, and 10-20 parts of euryale ferox.

2. The application according to claim 1, characterized in that, The traditional Chinese medicine composition is made from the following raw materials in parts by weight: 15 parts deer antler powder, 15 parts codonopsis, 15 parts salt-processed psoralea, 15 parts wine-processed cornus officinalis, 12 parts wine-processed cistanche deserticola, 6 parts prepared ginger, 24 parts bran-fried yam, 18 parts chicken blood vine, 6 parts prepared licorice root, 18 parts salvia miltiorrhiza, 15 parts vinegar-processed five-spice powder, and 15 parts euryale ferox.

3. The application according to claim 1, characterized in that, The dosage form of the traditional Chinese medicine composition is granules, decoction, powder, capsule, tablet or pill.

4. The application according to claim 1, characterized in that, The traditional Chinese medicine composition improves embryo implantation by activating the PPAR pathway, increasing the expression of PPARα and PPARγ, thereby improving or treating pregnancy outcomes in patients with thin endometrium undergoing frozen-thawed embryo transfer.

5. The application according to claim 1, characterized in that, The application is the use of a traditional Chinese medicine composition in combination with estrogen therapy in the preparation of a drug for improving pregnancy outcomes in frozen-thawed embryo transfer in patients with thin endometrium.

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