Objective To evaluate the impact of slow-freezing process on human ovarian tissue with the standard cryopreservation-thawing protocol of Fertility Protection Center of Beijing Obstetrics and Gynecology Hospital,Capital Medical University.Methods Ovarian tissues of 12 patients were divided into fresh ovarian tissue group(fresh group)and freezing-thawing ovarian tissue group(F-T group).The freezing-thawing protocol was the standard protocol in our center.The number and activity of follicle were examined with Hematoxylin-eosin(HE)staining and calcein-AM(calcein acetoxymethylester)staining,and the proliferation and apoptosis was evaluated with the immunohistochemical staining of Ki-67 and caspase-3.The expressions of apoptosis-related proteins such as caspase-3,bax and FasL between the two groups were compared with Western blotting.Results There were no statistically significant differences in follicle counting and follicle activity in ovarian tissues pre-and post-freezing-thawing(P>0.05),and the positive rate of Ki-67 in ovarian tissues after freezing-thawing was significantly lower than that in fresh ovarian tissues(P<0.05),and there was no statistically significant difference in the positive rate of caspase-3 between the two groups(P>0.05).The expression of caspase-3 protein in ovarian tissues after freezing-thawing was significantly higher than that in fresh ovarian tissues(P<0.05),while the expressions of other apoptosis-related proteins such as bax and FasL were not significantly different(P>0.05).Conclusion The standard cryopreservation-thawing regimen in our center can effectively maintain the follicle number,morphology,and activity in ovarian tissues.After freezing and thawing,the cell proliferation level is decreased.The expression of apoptosis-related proteins such as bax and FasL are not increased,and the expression of caspase-3 is relatively increased.These results suggest our freezing-thawing regimen is good for human ovarian tissue.

Objective To analyze the current status of bone mineral density(BMD)in women with early menopause and explore the correlations between BMD with age,body mass index(BMI),total body fat mass,spinal fat mass,femoral fat mass,follicle-stimulating hormone(FSH),estradiol(E2),and testosterone(T).Methods A total of 106 women with early menopause,who first visited the Department of Gynecological Endocrinology,Beijing Obstetrics and Gynecology Hospital,Capital Medical University from January 2023 to May 2025,were recruited after meeting the inclusion and exclusion criteria.The spinal BMD,femoral BMD,total body fat mass,femoral fat mass,and spinal fat mass were measured with dual-energy X-ray absorptiometry(DXA).The age,height,and weight of the patients were recorded,and serum levels of FSH,E2,and T were measured.Spearman correlation analysis was used to explore the correlations between spinal and femoral BMD and age,FSH,E2,T,BMI,total body fat content,femoral fat content,and spinal fat content.The patients were divided into three groups based on BMD:normal bone mass,osteopenia,and osteoporosis.Non-parametric Kruskal-Wallis H test was used to compare BMI,fat content,age,FSH,E2,and T levels among the three groups.Bonferroni-corrected Mann-Whitney U test was used for pairwise comparisons of significant differences.Multiple linear regression analysis was conducted to explore the influencing factors for femoral BMD T-score.Results Among the 106 patients with early menopause,30(28.3%)had normal bone mass,64(60.4%)had osteopenia,and 12(11.3%)had osteoporosis,with an average age of(43.99±0.16)years.Femoral BMD was positively correlated with BMI,total body fat mass,and spinal fat mass(all P<0.05).No significant differences were found in femoral fat mass,age,FSH,and T levels among the different bone mass groups,but,BMI,total body fat mass,spinal fat mass,and E2 were significantly different(P<0.05).BMI,total body fat mass,spinal fat mass and E2 were significantly higher in the normal bone mass group compared to the osteopenia group(P<0.05).The BMI of the normal bone mass group was significantly higher than that of the osteoporosis group(P<0.05).Multiple linear regression analysis showed that FSH were influencing factors for femoral BMD T-scores(P<0.05).Conclusion The osteoporosis is more prevalent in women with early menopause.Femoral BMD is positively correlated with BMI,total body fat mass and spinal fat content.Elevated FSH is a risk factor for reduced femoral BMD T-score.Attention should be paid to the risk of osteoporosis in women with early menopause,and individualized treatment plans should be developed to reduce the prevalence of osteoporosis.

Objective To compare ovulation recovery rate and metabolic indicators between ethinyl-estradiol/drospirenone(EE/DRSP)combined with orlistat and EE/DRSP alone in overweight/obese patients with polycystic ovary syndrome(PCOS).Methods This study was a randomized controlled clinical trial conducted based on the 2004 Rotterdam criteria.From October 2020 to December 2023,180 overweight/obese PCOS patients aged 20-40 were recruited from the Department of Department of Gynecological Endocrinology,Beijing Obstetrics and Gynecology Hospital,Capital Medical University.The patients were randomly divided into two groups in a 1∶2 ratio.Among them,60 patients received treatment with EE/DRSP(EE20 μg,DRSP 3 mg),while 120 patients received a combination treatment of EE/DRSP and orlistat(360 mg/d).The height,weight,waist circumference,hip circumference,and blood pressure of the patients were measured before treatment and after 12 weeks of treatment.Laboratory tests included measurements of follicle-stimulating hormone(FSH),luteinizing hormone(LH),fasting insulin(FINS),homeostasis model assessment of insulin resistance(HOMA-IR),free androgen index(FAI),alanine transaminase(ALT),aspartate transaminase(AST),gamma-glutamyl transferase(GGT),fasting plasma glucose(FPG),total cholesterol(TC),triglycerides(TG),high density lipoprotein-cholesterol(HDL-C),low density lipoprotein-cholesterol(LDL-C),lipoprotein(a)[Lp(a)],sex hormone-binding globulin(SHBG),total testosterone(T),and free testosterone(FT).After 12 weeks of treatment,the medication was discontinued,and natural ovulation was observed.Results After 12 weeks of treatment,the ovulation rate of the EE/DRSP combined with orlistat group reached 70.8%,while the natural ovulation rate of the EE/DRSP group alone was only 35%,indicating that the ovulation rate was significantly increased after EE/DRSP combined with orlistat treatment.After 12 weeks of treatment,both groups showed a significant decrease in total testosterone,free testosterone,and low-density lipoprotein levels(all P<0.05),and the decrease in BMI,waist circumference,fasting insulin,and HOMA-IR in the EE/DRSP combined with orlistat group was greater than that in the EE/DRSP group alone(P<0.05).After treatment,both groups showed a significant increase in high-density lipoprotein and triglyceride levels(all P<0.05),with no significant changes in total cholesterol and fasting blood glucose(all P>0.05).Conclusion After 12 weeks of treatment,EE/DRSP combined with orlistat can significantly improve the ovulation rate of PCOS patients.It is superior to EE/DRSP alone in reducing androgen levels,body weight,insulin resistance,and low-density lipoprotein levels.

Progesterone receptor membrane component 1(PGRMC1)is closely related to hormone therapy which belongs to the membrane-associated progesterone receptor(MAPR)family.A large number of in vitro experiments,in vivo animal experiments,clinical samples of breast cancer patients and blood studies showed that all synthetic progesterone(excluding natural progesterone and dydrogesterone)can promote the rapid proliferation of breast cancer cells overexpressing PGRMC1.In patients with breast cancer,PGRMC1 is significantly negatively correlated with tumor grade and prognosis,and PGRMC1 level in blood is positively correlated with PGRMC1 expression in breast cancer tissues,and PGRMC1 is superior to traditional tumor markers such as carcinoembryonic antigen(CEA),carbohydrate antigen(CA125),and CA153 in predicting early breast cancer.Therefore,PGRMC1 may serve as a predictive marker for identifying an elevated risk of breast cancer associated with menopausal hormone replacement therapy.

Objective To investigate the curative effect of letrozole combined with Zi Gui Nv Zhen Capsules on patients with polycystic ovary syndrome(PCOS)complicated by thin endometrial infertility.Methods 140 patients with PCOS who met the diagnosis of endometrial thin infertility(endometrial thickness<0.8 cm and no pregnancy for 1 year without contraception)were recruited and randomly assigned to the control group and the experimental group for 3 cycles of treatment.The control group was given letrozole 2.5 mg/d on the 3rd day of menstruation,estradiol valerate 2 mg/d on the 8-10th day of menstruation,and dydrogesterone 10 mg/d in sequence.The experimental group was treated with Zi Gui Nv Zhen Capsules on the basis of the control group.During 3 cycles of treatment,endometrial thickness(EMT),ovulation rate and cumulative pregnancy rate on the day of ovulation were examined.Results The EMT of the experiment group was significantly higher than that of the control group on the ovulation day of three cycles[(0.86±0.23)cm vs(0.71±0.15)cm;(0.83±0.26)cm vs(0.68±0.19)cm;(0.87±0.26)cm vs(0.71±0.11)cm],the cumulative pregnancy rate of experimental group was significantly higher than that of control group(44.3%vs 25.7%).There was no significant difference in ovulation rate between the two groups(P>0.05).Conclusion Our findings demonstrate that the combined use of Zi Gui Nv Zhen Capsule with basic conventional therapy significantly enhances endometrial thickness and improves pregnancy rates in PCOS patients with thin endometrium.

Objective To evaluate the impact of slow-freezing process on human ovarian tissue with the standard cryopreservation-thawing protocol of Fertility Protection Center of Beijing Obstetrics and Gynecology Hospital,Capital Medical University.Methods Ovarian tissues of 12 patients were divided into fresh ovarian tissue group(fresh group)and freezing-thawing ovarian tissue group(F-T group).The freezing-thawing protocol was the standard protocol in our center.The number and activity of follicle were examined with Hematoxylin-eosin(HE)staining and calcein-AM(calcein acetoxymethylester)staining,and the proliferation and apoptosis was evaluated with the immunohistochemical staining of Ki-67 and caspase-3.The expressions of apoptosis-related proteins such as caspase-3,bax and FasL between the two groups were compared with Western blotting.Results There were no statistically significant differences in follicle counting and follicle activity in ovarian tissues pre-and post-freezing-thawing(P>0.05),and the positive rate of Ki-67 in ovarian tissues after freezing-thawing was significantly lower than that in fresh ovarian tissues(P<0.05),and there was no statistically significant difference in the positive rate of caspase-3 between the two groups(P>0.05).The expression of caspase-3 protein in ovarian tissues after freezing-thawing was significantly higher than that in fresh ovarian tissues(P<0.05),while the expressions of other apoptosis-related proteins such as bax and FasL were not significantly different(P>0.05).Conclusion The standard cryopreservation-thawing regimen in our center can effectively maintain the follicle number,morphology,and activity in ovarian tissues.After freezing and thawing,the cell proliferation level is decreased.The expression of apoptosis-related proteins such as bax and FasL are not increased,and the expression of caspase-3 is relatively increased.These results suggest our freezing-thawing regimen is good for human ovarian tissue.

Objective To analyze the current status of bone mineral density(BMD)in women with early menopause and explore the correlations between BMD with age,body mass index(BMI),total body fat mass,spinal fat mass,femoral fat mass,follicle-stimulating hormone(FSH),estradiol(E2),and testosterone(T).Methods A total of 106 women with early menopause,who first visited the Department of Gynecological Endocrinology,Beijing Obstetrics and Gynecology Hospital,Capital Medical University from January 2023 to May 2025,were recruited after meeting the inclusion and exclusion criteria.The spinal BMD,femoral BMD,total body fat mass,femoral fat mass,and spinal fat mass were measured with dual-energy X-ray absorptiometry(DXA).The age,height,and weight of the patients were recorded,and serum levels of FSH,E2,and T were measured.Spearman correlation analysis was used to explore the correlations between spinal and femoral BMD and age,FSH,E2,T,BMI,total body fat content,femoral fat content,and spinal fat content.The patients were divided into three groups based on BMD:normal bone mass,osteopenia,and osteoporosis.Non-parametric Kruskal-Wallis H test was used to compare BMI,fat content,age,FSH,E2,and T levels among the three groups.Bonferroni-corrected Mann-Whitney U test was used for pairwise comparisons of significant differences.Multiple linear regression analysis was conducted to explore the influencing factors for femoral BMD T-score.Results Among the 106 patients with early menopause,30(28.3%)had normal bone mass,64(60.4%)had osteopenia,and 12(11.3%)had osteoporosis,with an average age of(43.99±0.16)years.Femoral BMD was positively correlated with BMI,total body fat mass,and spinal fat mass(all P<0.05).No significant differences were found in femoral fat mass,age,FSH,and T levels among the different bone mass groups,but,BMI,total body fat mass,spinal fat mass,and E2 were significantly different(P<0.05).BMI,total body fat mass,spinal fat mass and E2 were significantly higher in the normal bone mass group compared to the osteopenia group(P<0.05).The BMI of the normal bone mass group was significantly higher than that of the osteoporosis group(P<0.05).Multiple linear regression analysis showed that FSH were influencing factors for femoral BMD T-scores(P<0.05).Conclusion The osteoporosis is more prevalent in women with early menopause.Femoral BMD is positively correlated with BMI,total body fat mass and spinal fat content.Elevated FSH is a risk factor for reduced femoral BMD T-score.Attention should be paid to the risk of osteoporosis in women with early menopause,and individualized treatment plans should be developed to reduce the prevalence of osteoporosis.

Objective To compare ovulation recovery rate and metabolic indicators between ethinyl-estradiol/drospirenone(EE/DRSP)combined with orlistat and EE/DRSP alone in overweight/obese patients with polycystic ovary syndrome(PCOS).Methods This study was a randomized controlled clinical trial conducted based on the 2004 Rotterdam criteria.From October 2020 to December 2023,180 overweight/obese PCOS patients aged 20-40 were recruited from the Department of Department of Gynecological Endocrinology,Beijing Obstetrics and Gynecology Hospital,Capital Medical University.The patients were randomly divided into two groups in a 1∶2 ratio.Among them,60 patients received treatment with EE/DRSP(EE20 μg,DRSP 3 mg),while 120 patients received a combination treatment of EE/DRSP and orlistat(360 mg/d).The height,weight,waist circumference,hip circumference,and blood pressure of the patients were measured before treatment and after 12 weeks of treatment.Laboratory tests included measurements of follicle-stimulating hormone(FSH),luteinizing hormone(LH),fasting insulin(FINS),homeostasis model assessment of insulin resistance(HOMA-IR),free androgen index(FAI),alanine transaminase(ALT),aspartate transaminase(AST),gamma-glutamyl transferase(GGT),fasting plasma glucose(FPG),total cholesterol(TC),triglycerides(TG),high density lipoprotein-cholesterol(HDL-C),low density lipoprotein-cholesterol(LDL-C),lipoprotein(a)[Lp(a)],sex hormone-binding globulin(SHBG),total testosterone(T),and free testosterone(FT).After 12 weeks of treatment,the medication was discontinued,and natural ovulation was observed.Results After 12 weeks of treatment,the ovulation rate of the EE/DRSP combined with orlistat group reached 70.8%,while the natural ovulation rate of the EE/DRSP group alone was only 35%,indicating that the ovulation rate was significantly increased after EE/DRSP combined with orlistat treatment.After 12 weeks of treatment,both groups showed a significant decrease in total testosterone,free testosterone,and low-density lipoprotein levels(all P<0.05),and the decrease in BMI,waist circumference,fasting insulin,and HOMA-IR in the EE/DRSP combined with orlistat group was greater than that in the EE/DRSP group alone(P<0.05).After treatment,both groups showed a significant increase in high-density lipoprotein and triglyceride levels(all P<0.05),with no significant changes in total cholesterol and fasting blood glucose(all P>0.05).Conclusion After 12 weeks of treatment,EE/DRSP combined with orlistat can significantly improve the ovulation rate of PCOS patients.It is superior to EE/DRSP alone in reducing androgen levels,body weight,insulin resistance,and low-density lipoprotein levels.

Progesterone receptor membrane component 1(PGRMC1)is closely related to hormone therapy which belongs to the membrane-associated progesterone receptor(MAPR)family.A large number of in vitro experiments,in vivo animal experiments,clinical samples of breast cancer patients and blood studies showed that all synthetic progesterone(excluding natural progesterone and dydrogesterone)can promote the rapid proliferation of breast cancer cells overexpressing PGRMC1.In patients with breast cancer,PGRMC1 is significantly negatively correlated with tumor grade and prognosis,and PGRMC1 level in blood is positively correlated with PGRMC1 expression in breast cancer tissues,and PGRMC1 is superior to traditional tumor markers such as carcinoembryonic antigen(CEA),carbohydrate antigen(CA125),and CA153 in predicting early breast cancer.Therefore,PGRMC1 may serve as a predictive marker for identifying an elevated risk of breast cancer associated with menopausal hormone replacement therapy.

Objective To investigate the curative effect of letrozole combined with Zi Gui Nv Zhen Capsules on patients with polycystic ovary syndrome(PCOS)complicated by thin endometrial infertility.Methods 140 patients with PCOS who met the diagnosis of endometrial thin infertility(endometrial thickness<0.8 cm and no pregnancy for 1 year without contraception)were recruited and randomly assigned to the control group and the experimental group for 3 cycles of treatment.The control group was given letrozole 2.5 mg/d on the 3rd day of menstruation,estradiol valerate 2 mg/d on the 8-10th day of menstruation,and dydrogesterone 10 mg/d in sequence.The experimental group was treated with Zi Gui Nv Zhen Capsules on the basis of the control group.During 3 cycles of treatment,endometrial thickness(EMT),ovulation rate and cumulative pregnancy rate on the day of ovulation were examined.Results The EMT of the experiment group was significantly higher than that of the control group on the ovulation day of three cycles[(0.86±0.23)cm vs(0.71±0.15)cm;(0.83±0.26)cm vs(0.68±0.19)cm;(0.87±0.26)cm vs(0.71±0.11)cm],the cumulative pregnancy rate of experimental group was significantly higher than that of control group(44.3%vs 25.7%).There was no significant difference in ovulation rate between the two groups(P>0.05).Conclusion Our findings demonstrate that the combined use of Zi Gui Nv Zhen Capsule with basic conventional therapy significantly enhances endometrial thickness and improves pregnancy rates in PCOS patients with thin endometrium.