Therapeutic effects of curculigoside on cyclophosphamide-induced premature ovarian failure in mice.

OBJECTIVE: The main purpose of this study was to elucidate the anti-apoptotic effects of curculigoside (CUR) on ovarian granulosa cells (GCs) in a mouse model of cyclophosphamide (CTX)-induced premature ovarian failure (POF). METHOD: Intraperitoneal injection of CTX (100 mg/kg body weight) induced POF in mice. Thirty-six female mice were divided into six groups: blank group; POF model group; low-dose CUR group; medium-dose CUR group; high-dose CUR group; and estradiol benzoate group. Mice were orally administered for 28 consecutive days. Twenty-four hours after the completion of treatment, mice were weighed and euthanized, and blood was collected from the eyeball under anesthesia. The ovaries were surgically separated and weighed, and the ovarian index was calculated. Hematoxylin-eosin (HE) staining was used to observe follicular development and corpus luteum morphology in the ovaries. Serum levels of follicle stimulating hormone (FSH), anti-Müllerian hormone (AMH) and estradiol (E2) were measured. Superoxide dismutase (SOD) activity, glutathione peroxidase (GSH-Px) content and malondialdehyde (MDA) levels in ovarian tissue were determined. The GC apoptosis level was measured. Western blotting was used to detect protein expression levels of Beclin-1, LC3, P62, AKT, p-AKT, mTOR and p-mTOR in the ovaries. RESULTS: The results showed that CUR can improve body weight and ovarian index; promote follicular development and reduce follicular atresia; improve FSH, AMH and E2 levels; downregulate MDA levels and restore antioxidant enzyme activity; inhibit the autophagy level; activate the AKT/mTOR signaling pathway; and alleviate GC apoptosis. CONCLUSION: CUR improves POF by activating the AKT/mTOR signaling pathway, inhibiting autophagy and alleviating GC apoptosis.

Analysis of the potential regulatory mechanisms of female and latent genital tuberculosis affecting ovarian reserve function using untargeted metabolomics.

Female and latent genital tuberculosis (FGTB and LGTB) in young women may lead to infertility by damaging ovarian reserve function, but the regulatory mechanisms remain unclear. In this study, we investigated the effects of FGTB and LGTB on ovarian reserve function and potential regulatory mechanisms by untargeted metabolomics of follicular fluid, aiming to provide insights for the clinical management and treatment approaches for afflicted women. We recruited 19 patients with FGTB, 16 patients with LGTB, and 16 healthy women as a control group. Clinical data analysis revealed that both the FGTB and LGTB groups had significantly lower ovarian reserve marker levels compared to the control group, including lower anti-Müllerian hormone levels (FGTB: 0.82 [0.6, 1.1] µg/L; LGTB: 1.57 [1.3, 1.8] µg/L vs. control: 3.29 [2.9, 3.5] µg/L), reduced antral follicular counts (FGTB: 6 [5.5, 9.5]; LGTB: 10.5 [7, 12.3] vs. control: 17 [14.5, 18]), and fewer retrieved oocytes (FGTB: 3 [2, 5]; LGTB: 8 [4, 8.3] vs. control: 14.5 [11.5, 15.3]). Conversely, these groups exhibited higher ovarian response marker levels, such as longer gonadotropin treatment days (FGTB: 12 [10.5, 12.5]; LGTB: 11 [10.8, 11.3] vs. control: 10 [8.8, 10]) and increased gonadotropin dosage requirements (FGTB: 3300 [3075, 3637.5] U; LGTB: 3037.5 [2700, 3225] U vs. control: 2531.25 [2337.5, 2943.8] U). All comparisons were statistically significant at P < 0.05. The results suggested that FGTB and LGTB have adverse effects on ovarian reserve and response. Untargeted metabolomic analysis identified 92 and 80 differential metabolites in the control vs. FGTB and control vs. LGTB groups, respectively. Pathway enrichment analysis revealed significant alterations in metabolic pathways in the FGTB and LGTB groups compared to the control group (P < 0.05), with specific changes noted in galactose metabolism, biotin metabolism, steroid hormone biosynthesis, and nicotinate and nicotinamide metabolism in the FGTB group, and caffeine metabolism, primary bile acid biosynthesis, steroid hormone biosynthesis, and glycerophospholipid metabolism in the LGTB group. The analysis of metabolic levels has revealed the potential mechanisms by which FGTB and LGTB affect ovarian reserve function, namely through alterations in metabolic pathways. The study emphasizes the importance of comprehending the metabolic alterations associated with FGTB and LGTB, which is of considerable relevance for the clinical management and therapeutic approaches in afflicted women.

HGF-modified human umbilical cord mesenchymal stem cells rescue impaired ovarian reserve function in chemotherapy-induced POI rats by improving angiogenesis while decreasing apoptosis and fibrosis in the ovary.

Complications caused by Primary ovarian insufficiency (POI), including infertility, osteoporosis, cardiovascular diseases and depression, severely affect the life quality of female patients. Although hormone replacement therapy (HRT) can alleviate some long-term complications, there is still no standard treatment for the restoration of ovarian reserve function. Currently, human umbilical cord mesenchymal stem cells (HUCMSC) transplantation showed considerable treatment effect for POI in both rat model and clinic. To improve the effectiveness of naïve HUCMSC (HUCMSC-Null) treatments on POI, an exogenous gene hepatocyte growth factor (HGF) which promotes follicular angiogenesis in POI ovaries was used to modify HUCMSC. Subsequently, HGF-overexpressed HUCMSC (HUCMSC-HGF) was transplanted into the ovaries of chemotherapy-induced POI Sprague-Dawley (SD) rats to observe the effectiveness on POI improvement and its related mechanisms. Our results showed that when compared with POI and HUCMSC-Null treatment group, HUCMSC-HGF significantly improved ovarian reserve function in POI group, which might be attributed to the decrease of ovarian tissue fibrosis and granulosa cells (GCs) apoptosis, and the increase of ovarian angiogenesis mediated by HGF over-expression. The findings suggest that HGF-modified HUCMSC may present a more superior capacity than HUCMSC alone for the rescue of ovarian reserve function in POI.

Relationship between thyroid antibody levels and ovarian reserve function in infertile chinese women with normal thyroid-stimulating hormone.

BACKGROUND: To analyze the relationship of thyroid peroxidase antibody and thyroid globulin antibody levels with ovarian reserve function in infertile women. METHODS: The data of 721 infertile patients who visited the hospital from January 2019 to September 2022 and whose thyroid-stimulating hormone (TSH), free triiodothyronine (FT3), and free thyroxine (FT4) levels were in the normal range, were retrospectively analyzed. These patients were divided into two sets of three groups-the negative group, the 2.6 IU/ml ~ 100 IU/ml group and the TPOAb > 100 IU/ml group according to the TPOAb (thyroid peroxidase antibody) level, or the TgAb (anti-thyroglobulin antibody) negative group, the 14.58 IU/ml ~ 100 IU/ml group and the TgAb > 100 IU/ml group according to the TgAb level. They were compared for differences in ovarian reserve function index and thyroid hormone levels and analyzed for the relationship among thyroid antibody levels, ovarian reserve function, and thyroid hormone levels. RESULTS: When TSH > 2.5 mIU/L, the bFSH (basal follicle stimulating hormone) level in the TPOAb > 100 IU/ml group (9.10 ± 1.16 IU/L) was significantly higher than that in the TPOAb negative group (8.12 ± 1.97 IU/L) and the 2.6 IU/ml ~ 100 IU/ml group (7.90 ± 1.48 IU/L) (P < 0.05); when TSH ≤ 2.5 mIU/L, there were no statistically significant differences in the bFSH and AFC (antral follicle count) number at different TPOAb levels. Whether TSH ≤ 2.5 mIU/L or TSH > 2.5 mIU/L, there were no statistically significant differences in the bFSH and AFC number at different TgAb levels (P > 0.05). FT3/FT4 ratio in the TPOAb 2.6 IU/ml ~ 100 IU/ml group and the > 100 IU/ml group was significantly lower than in the negative group. FT3/FT4 ratio in the TgAb 14.58 ~ 100 IU/ml group and the > 100 IU/ml group was also significantly lower than in the TgAb negative group (P < 0.05). TSH level in the TPOAb > 100 IU/ml group was significantly higher than in the 2.6 ~ 100 IU/ml group and the TPOAb negative group, but there were no statistically significant differences among different TgAb groups. CONCLUSIONS: When TPOAb > 100 IU/ml and TSH > 2.5 mIU/L, it may affect the ovarian reserve function in infertile patients, and the mechanism may be associated with increased TSH and the imbalance of FT3/FT4 ratio caused by the increase of TPOAb.

Multiple Protocols Combined with Hyperbaric Oxygen Therapy on the Maintenance of Ovarian Function in Patients After Ovarian Cystectomy.

Objective: This study aims to explore the effect of adjuvant hyperbaric oxygen therapy on ovarian function after laparoscopic ovarian cystectomy. Methods: A total of 60 patients with ovarian cysts treated at our hospital from January 2018 to August 2020 were enrolled. According to the different treatment modalities, the patients were divided into the control and observation groups. Patients in both groups underwent laparoscopic ovarian cystectomy with oral administration of Chinese patent medicine Kuntai capsules after surgery. Hyperbaric oxygen therapy was added to patients in the observation group in addition to the treatment in the control group. The anti-Müllerian hormone (AMH), follicle-stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E2), and antral follicle count (AFC) serum levels were detected in both groups before the operation and at the first and third menstrual cycles postoperatively to evaluate ovarian function. Results: At the first and third menstrual cycles after surgery, the AMH, E2, and AFC serum levels in the two groups were significantly lower than before surgery, and the FSH and LH serum levels were higher than before surgery. The differences were statistically significant (P < 0.05). After the operation, AMH, E2, and AFC serum levels in the observation group were significantly higher than in the control group. FSH and LH serum levels were significantly lower than in the control group, and the differences were statistically significant (P < 0.05). Conclusion: For patients undergoing laparoscopic ovarian cystectomy, the adjuvant hyperbaric oxygen therapy could significantly improve the postoperative ovarian reserve function with remarkable effects.

Clinical observation of Kuntai capsule combined with Fenmotong in treatment of decline of ovarian reserve function.

BACKGROUND: Decreased ovarian reserve function is an ovarian hypofunction disease that occurs in women before 40 years of age, leading to a decline in fertility and perimenopausal symptoms, such as irregular menstruation, amenorrhea, infertility, decreased libido, and autonomic nervous dysfunction. Fenmatong (FMT) is a compound mixture of estradiol tablets and estradiol didroxyprogesterone tablets, which can improve ovarian reserve function by supplementation of exogenous estrogen. However, this treatment has also been shown to cause breast pain, gastrointestinal discomfort, irregular vaginal bleeding, and changes in sexual desire. In severe cases, FMT can promote the development of breast cancer, endometrial cancer, and venous embolic disease. AIM: To observe the effects of Kuntai capsules and FMT on endocrine indexes and uterine artery blood circulation in patients with decreased ovarian reserve function. METHODS: Patients (130) with decreased ovarian reserve function, who were treated in our hospital from May 2018 to May 2020, were divided into two groups: The FMT group, in which patients were treated with FMT, and the observation group, in which patients were treated with Kuntai capsules. Chinese medicine symptom scores, uterine artery blood flow parameters, ovarian ultrasound test indexes, pictorial blood loss assessment chart (PBAC) scores, and hormone levels were recorded, and total effective rates were calculated for both groups. RESULTS: The total effective rate in the observation group was higher than that in the FMT group (P < 0.05).After treatment, primary symptoms, including low menstrual volume, delayed menstruation, red color and thick consistency of menses, dizziness, palpitation, weakness at the waist and knee, insomnia and excessive dreaming, irritability, and dryness and astringency of the pudendal canal in the observation group decreased, and scores for primary and secondary symptoms in the observation group were significantly lower than those in the FMT group (P < 0.05).The systolic peak flow rate (PSV), end-diastolic flow rate (EDV), ovarian diameter, sinus follicle count, and resistance index (RI) of the uterine arteries in the observation group and FMT group increased after treatment. Notably, the PSV, EDV, ovarian diameter, and antral follicle count in the observation group were higher than those in the FMT group, whereas the RI in the observation group was lower than that in the FMT group (P < 0.05).The PBAC scores in the observation and FMT groups increased after treatment, with that in the observation group becoming significantly higher than that in the FMT group (P < 0.05). After treatment, estradiol (E2) and anti-Mullerian hormone (AMH) levels increased, whereas follicle-stimulating hormone (FSH) levels decreased in the observation group and FMT group; E2 and AMH levels became significantly higher and FSH levels became significantly lower in the observation group than in the FMT group (P < 0.05). CONCLUSION: Compared with FMT, Kuntai capsules promoted uterine artery blood circulation, improved menstruation, relieved symptoms, regulated endocrine function, and improved curative effects.

Development of an optimal protocol for isolation and purification of human granulosa cells in patients with different ovarian reserves.

The aim of the current study was to develop an accurate and reproducible method for isolation of granulosa cells (GCs) in patients with different ovarian reserves. The cells of healthy individuals and patients with polycystic ovary syndrome (PCOS) were isolated using a modified two-step Percoll density gradient centrifugation. The cells of patients with poor ovarian response (POR) were isolated using a one-step method suitable for samples containing few cells. Cells extracted using these purification techniques were compared regarding cell yield, viability and purity using immunocytochemistry, flow cytometry, Cell Counting Kit-8, western blotting and RNA integrity analysis. The purity and activity of the cells in the POR group were comparable with those in the healthy and PCOS groups and no statistically significant differences were identified. Furthermore, isolated cells analyzed for RNA integrity indicated good quality RNA and presented an RNA integrity number of 8-10, demonstrating that the technique enabled the isolation of GCs from different types of patients. Thus, a reliable and reproducible technique for the isolation of pure GCs with high yield is described in the present study. This protocol provides an efficient method targeted to patients with different ovarian reserve functions that enables the preparation of GCs for evaluating their molecular functions.

Clinical significance of combined detection of anti-Mullerian hormone and follicular output rate in women of late reproductive age.

OBJECTIVE: This study was designed to explore the clinical significance of anti-Mullerian hormone (AMH) combined with follicular output rate (FORT) in women of late reproductive age. METHODS: A total of 258 women (age range: 35-45 years old) who underwent pre-pregnancy examination in our hospital were collected as the research group (RG), among whom 184 were treated with in vitro fertilization-embryo transfer (IVF-ET). Concurrently, 126 women aged 24-30 years who came to our hospital for pre-pregnancy examination were enrolled as the control group (CG). AMH and FORT were detected and compared between the two groups to analyze the clinical significance of the two in women of late reproductive age. RESULTS: Compared with the CG, AMH was decreased statistically in the RG (P<0.05). AMH was statistically higher in the regular menstrual group than in the menstrual disorder group (P<0.05), and FORT was statistically higher in the pregnancy group in comparison with the non-pregnancy group (P<0.05). AMH decreased with age (P<0.05), while FORT did not correlate with any notable difference among the three subgroups (P>0.05). High, medium and low AMH groups showed no significant difference in the number of retrieved oocytes and transplantable embryos, as well as FORT (P<0.05). A lower AMH level, was correlated with fewer number of retrieved oocytes and transplantable embryos, and higher the FORT level. Significant differences were present among the high, middle and low FORT groups regarding the number of retrieved oocytes and transplantable embryos, the clinical pregnancy rate and AMH level (P<0.05). The lower the level of FORT was, the less the number of retrieved oocytes and transplantable embryos was, the lower clinical pregnancy rate was, and the higher the AMH level was. CONCLUSIONS: AMH decreases gradually in women with an increase of age, and FORT can effectively predict pregnancy outcome. AMH detection combined FORT is of great significance in predicting the ovarian reserve function in women of late reproductive age.

The effects of multiple controlled ovarian hyperstimulation over a 2-year period on ovarian reserve and reactivity: a retrospective clinical study.

BACKGROUND: To evaluate the effects of controlled ovarian hyperstimulation (COH) on ovarian reserve function during in vitro fertilization and embryo transfer (IVF-ET). METHODS: From August 2018 to August 2020, the medical records of patients who received IVF-ET in the Department of Reproductive Medicine, Beijing Gynaecology and Obstetrics Hospital, Capital Medical University were analyzed retrospectively. Among them, 372 patients received 2 cycles of COH, 54 patients received 3 cycles, and 13 patients received 4 cycles. The levels of follicle-stimulating hormone (FSH), the number of antral follicles, levels of anti-Müllerian hormone (AMH), the total amount of gonadotropin (GN), the time of ovulation induction, the number of eggs obtained, the number of available embryos, and the number of high-quality embryos cycles were compared in different treatment. RESULTS: The age of female patients did not significantly affect the levels of AMH or FSH during menstruation, nor the number of antral follicles before ovulation induction (P>0.05). However, with an increase in age, an increase in the number of controlled COH cycles was observed. In patients who underwent 2 COH cycles, the number of high-quality embryos in the second cycle increased significantly compared to the first cycle (P<0.05). However, there were no significant differences in the ovulation induction time, the number of eggs, the GN dosage, and the number of available embryos (P>0.05). In patients with 3 treatment cycles, the GN dose used in the third cycle was significantly lower than that used in the first cycle (P<0.05). There were no significant differences in the ovulation induction time, the number of eggs obtained, and the quality of embryos (P>0.05). In patients with 4 treatment cycles, significant differences were observed in the ovulation induction time between the first and the fourth controlled COH cycle (P<0.05). However, no significant differences were detected in GN dosage, ovulation induction time, the number of eggs obtained, the number of available embryos, and the quality of embryos (P>0.05). CONCLUSIONS: Ovarian reserve function was not significantly affected in patients with up to 4 ovarian stimulation cycles.

The effect of adenomyosis on IVF after long or ultra-long GnRH agonist treatment.

RESEARCH QUESTION: Does adenomyosis affect IVF independent of decreased ovarian reserve, and what are the characteristics and IVF outcome of the ultra-long gonadotrophin-releasing hormone (GnRH) agonist protocol in adenomyosis? DESIGN: Observational cohort study of three groups of patients undergoing first cycle of IVF treatment with normal ovarian reserve: (A) 362 patients with adenomyosis using the ultra-long GnRH agonist protocol; (B) 127 patients with adenomyosis using the long GnRH agonist protocol; (C) 3471 patients with tubal infertility using the long GnRH agonist protocol. RESULTS: Compared with groups B and C, the number of oocytes retrieved in group A decreased, and the gonadotrophin dosage and duration in group A were higher (P < 0.001). In long GnRH agonist treatment, clinical pregnancy rate (OR 0.492, 95% CI 0.327 to 0.742, P < 0.001), implantation rate (OR 0.527, 95% CI 0.350 to 0.794, P = 0.002) and live birth rate (OR 0.442, 95% CI 0.291 to 0.673, P < 0.001) decreased and miscarriage rate (OR 3.078, 95% CI 1.593 to 5.948, P < 0.001) increased in adenomyosis patients compared with tubal infertility. For adenomyosis patients, clinical pregnancy rate (OR 1.925, 95% CI 1.137 to 3.250, P = 0.015), implantation rate (OR 1.694, 95% CI 1.006 to 2.854, P = 0.047) and live birth rate (OR 1.704, 95% CI 1.012 to 2.859, P = 0.044) increased in the ultra-long GnRH agonist treatment compared with long GnRH agonist treatments. CONCLUSION: Adenomyosis could negatively affect IVF outcomes independent of ovarian reserve after long GnRH agonist protocol. Patients with adenomyosis following the ultra-long GnRH agonist protocol could have a better pregnancy outcome than those following the long GnRH agonist protocol.

[Effect of electroacupuncture combined with Yuyin pill on sex hormone and Th2 cytokines in patients of decreased ovarian reserve function with liver-kidney yin deficiency].

OBJECTIVE: To observe the effect of electroacupuncture (EA) combined with Yuyin pill on clinical symptoms, levels of serum sex hormone and Th2 cytokines in patients of decreased ovarian reserve function (DOR) with liver-kidney yin deficiency, and to compare the efficacy between EA combined with Yuyin pill and Yuyin pill alone. METHODS: Sixty patients with DOR were randomly divided into an observation group (30 cases, 2 cases dropped off) and a control group (30 cases, 1 case dropped off). The patients in the control group were treated with Yuyin pill, 1 pill each time, 3 times a day. Based on the treatment of the control group, the patients in the observation group were additionally treated with acupuncture at Guanyuan (CV 4), Zhongji (CV 3), Guilai (ST 29), Zigong (EX-CA 1), Zusanli (ST 36), Sanyinjiao (SP 6), Taixi (KI 3) and Taichong (LR 3); EA was applied at bilateral Zusanli (ST 36) and Sanyinjiao (SP 6), with continuous wave, in frequency of 20 Hz and current intensity of 1 to 4 mA, for 20 min. The treatment was given 3 times a week. All the patients terminated treatment during menstrual period, and the treatment was given for 3 continuous menstrual cycles. The menstrual condition score and systemic symptom score were compared between the two groups before and after treatment. The levels of serum sex hormones on 2nd to 3rd day of menstruation, including follicle stimulating hormone (FSH), luteinizing hormone (LH) and estradiol (E2), and the serums levels of interleukin (IL) -4 and IL-10 secreted by Th2 cytokines were compared between the two groups before and after treatment. RESULTS: After the treatment, the menstruation condition scores and systemic symptom scores in the two groups were reduced (P<0.05), and the scores in the observation group were lower than those in the control group (P<0.05). After the treatment, the levels of serum FSH, LH and FSH/LH were reduced (P<0.05), and the E2 levels were increased in the two groups (P<0.05), and the levels of FSH, LH in the observation group were lower than those in the control group (P<0.05), and the E2 level was higher than that in the control group (P<0.05). After the treatment, the levels of serum IL-4 and IL-10 in the two groups were increased (P<0.05), and the levels of IL-4 and IL-10 in the observation group were higher than those in the control group (P<0.05). CONCLUSION: EA combined with Yuyin pill could significantly improve menstruation, systemic symptoms and serum sex hormone levels in patients of decreased ovarian reserve function with liver-kidney yin deficiency, which may restore ovarian function by up-regulating the expression of Th2 cytokines.

Human umbilical cord mesenchymal stem cells improve the reserve function of perimenopausal ovary via a paracrine mechanism.

BACKGROUND: Human umbilical cord mesenchymal stem cells (hUCMSCs) are a type of pluripotent stem cell which are isolated from the umbilical cord of newborns. hUCMSCs have great therapeutic potential. We designed this experimental study in order to investigate whether the transplantation of hUCMSCs can improve the ovarian reserve function of perimenopausal rats and delay ovarian senescence. METHOD: We selected naturally aging rats confirmed by vaginal smears as models of perimenopausal rats, divided into the control group and the treatment group, and selected young fertile female rats as normal controls. hUCMSCs were transplanted into rats of the treatment group through tail veins. Enzyme-linked immunosorbent assay (ELISA) detected serum levels of sex hormones, H&E staining showed ovarian tissue structure and allowed follicle counting, immunohistochemistry and western blot analysis revealed ovarian expression of hepatocyte growth factor (HGF), vascular endothelial cell growth factor (VEGF), and insulin-like growth factor-1 (IGF-1), polymerase chain reaction (PCR) and western blot analysis revealed hUCMSCs expression of HGF, VEGF, and IGF-1. RESULTS: At time points of 14, 21, and 28 days after hUCMSCs transplantation, estradiol (E2) and anti-Müllerian hormone (AMH) increased while follicle-stimulating hormone (FSH) decreased; ovarian structure improved and follicle number increased; ovarian expression of HGF, VEGF, and IGF-1 protein elevated significantly. Meanwhile, PCR and western blot analysis indicated hUCMSCs have the capacity of secreting HGF, VEGF, and IGF-1 cytokines. CONCLUSIONS: Our results suggest that hUCMSCs can promote ovarian expression of HGF, VEGF, and IGF-1 through secreting those cytokines, resulting in improving ovarian reserve function and withstanding ovarian senescence.

[Effects of ginger-separated moxibustion at Baliao points combined with Bushen Huoxue formula on patients with decreased ovarian reserve function].

OBJECTIVE: To compare the efficacy differences between ginger-separated moxibustion at Baliao points combined with Bushen Huoxue formula and Bushen Huoxue formula alone on patients with decreased ovarian reserve function. METHODS: Fifty patients of decreased ovarian reserve function were randomly divided into an observation group and a control group, 25 cases in each one. The patients in the observation group were treated with ginger-separated moxibustion at Baliao points combined with Bushen Huoxue formula; the moxibustion was given for 1.5 h, once every seven days, and no treatment was given during menstrual period. The patients in the control group were treated with Bushen Huoxue formula. One-month treatment was taken as one treatment course, and totally three courses were given. The change of follicle-stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E2), anti-mullerian hormone (AMH), antral follicle count (AFC), peak systolic velocity (PSV), resistance index (RI) were observed before and after treatment in the two groups. RESULTS: After treatment, the FSH, FSH/LH and RI were significantly lowered, but the E2, AFC, PSV were significantly increased in the two groups (all P<0.05); the FSH, FSH/LH and E2 in the observation group were lower and AFC was higher than those in the control group (all P<0.05). CONCLUSION: The ginger-separated moxibustion at Baliao points combined with Bushen Huoxue formula are superior to Bushen Huoxue formula alone in improving ovarian reserve function.