OBJECTIVES: Platelet-rich plasma (PRP) treatment has been reported to improve ovarian function in women, but the relationship between the dose and frequency of PRP treatment and its therapeutic effect remains unclear. This study aims to evaluate the efficacy of single and double PRP treatments for diminished ovarian reserve (DOR). METHODS: A total of 65 patients treated at the Reproductive Center of Xiangtan Central Hospital from September 2020 to October 2022 were randomly divided into 4 groups: A single PRP treatment group (21 patients, PRP treatment once), a double PRP treatment group (15 patients, PRP treatment twice), a blank control group (15 patients), and an artificial cycle control group (14 patients, treated with estrogen and progesterone). The differences in baseline follicle-stimulating hormone (FSH), antral follicle count (AFC), ovarian volume, number of oocytes retrieved, number of good quality embryos, and pregnancy outcomes before and after treatment were compared among the 4 groups. RESULTS: Compared to before treatment, both single and double PRP treatment groups showed a significant reduction in FSH, and an increase in AFC, ovarian volume, numbers of oocytes retrieved, and number of MII oocytes (all P<0.05). Compared to the blank control group, the single and double PRP groups showed a decrease in FSH, with an increase in AFC, number of oocytes retrieved, and number of MII oocytes (all P<0.05). The rates of good quality embryos, clinical pregnancy rate, and live birth rate in the single and double PRP groups were higher than those in the blank control group and artificial cycle control groups, but the differences were not statistically significant (all P>0.05). Compared to the single PRP treatment group, the double PRP group had lower FSH and higher AFC, but the differences were not statistically significant (both P>0.05). CONCLUSIONS Both single and double PRP ovarian injections can effectively improve ovarian reserve function in DOR patients and enhance ovarian response. Compared to single PRP ovarian injection, double PRP ovarian injection shows a trend of better improvement in ovarian reserve function.
Humans ; Female ; Platelet-Rich Plasma ; Ovarian Reserve/physiology* ; Adult ; Pregnancy ; Follicle Stimulating Hormone/blood* ; Pregnancy Rate ; Ovary ; Ovulation Induction/methods* ; Ovarian Follicle ; Oocyte Retrieval

OBJECTIVE: Hyperstimulation methods are broadly used for in vitro fertilization (IVF) in patients with infertility; however, the side effects associated with these therapies, such as ovarian hyperstimulation syndrome (OHSS), have not been well studied. N-glycoproteomes are subproteomes used for the remote sensing of ovarian stimulation in follicular growth. Glycoproteomic variation in human follicular fluid (hFF) has not been evaluated. In this study, we aimed to identify and quantify the glycoproteomes and N-glycoproteins (N-GPs) in natural and stimulated hFF using label-free nano-liquid chromatography/electrospray ionization-quad time-of-flight mass spectrometry. METHODS: For profiling of the total proteome and glycoproteome, pooled protein samples from natural and stimulated hFF samples were selectively isolated using hydrazide chemistry to obtain the total proteomes and glycoproteomes. N-GPs were validated by the consensus sequence N-X-S/T (92.2% specificity for the N-glycomotif at p<0.05). All data were compared between natural versus hyperstimulated hFF samples. RESULTS: We detected 41 and 44 N-GPs in the natural and stimulated hFF samples, respectively. Importantly, we identified 11 N-GPs with greater than two-fold upregulation in stimulated hFF samples compared to natural hFF samples. We also validated the novel N-GPs thyroxine-binding globulin, vitamin D-binding protein, and complement proteins C3 and C9. CONCLUSION: We identified and classified N-GPs in hFF to improve our understanding of follicular physiology in patients requiring assisted reproduction. Our results provided important insights into the prevention of hyperstimulation side effects, such as OHSS.
Chemistry ; Complement System Proteins ; Consensus Sequence ; Female ; Fertilization in Vitro* ; Follicular Fluid* ; Humans* ; In Vitro Techniques* ; Infertility ; Mass Spectrometry ; Ovarian Hyperstimulation Syndrome ; Ovulation Induction ; Physiology ; Proteome ; Proteomics ; Reproduction ; Sensitivity and Specificity ; Thyroxine-Binding Globulin ; Up-Regulation ; Vitamin D-Binding Protein

This study investigated the molecular markers of DS-1-47, a component of an implantation- promoting traditional Chinese medicine consisting of Astragalus mongholicus, Atractylodes macrocephala, Scutellaria baicalensis and Dipsacales, in an attempt to clarify the molecular mechanism and action targets of DS-1-47. Controlled ovarian stimulation (COS) method was used to establish the implantation dysfunction models of mice. Animals were divided into normal pregnant group, COS model group and DS-1-47 group. Laser capture microdissection-double dimensional electrophoresis-mass spectrum (LCM-DE-MS) was used to analyze the uterine protein molecules that were possibly involved in the promotion of implantation. Twenty-three proteins in DS-1-47 group were significantly changed as compared to those in COS model group, with 7 proteins down-regulated and 16 proteins up-regulated. Except for some constituent proteins, the down-regulated proteins included collagen α-1 (VI) chain, keratin 7, keratin 14, myosin regulatory light chain 12B, myosin light polypeptide 9, heat shock protein β-7, and C-U-editing enzyme APOBEC-2; the up-regulated proteins included apolipoprotein A-I, calcium regulated protein-3, proliferating cell nuclear antigen, L-xylulose reductase, and calcium binding protein. These 23 proteins that were regulated by DS-1-47 represented a broad diversity of molecule functions. The down-regulated proteins were associated with stress and immune response, and those up-regulated proteins were related to proliferation. It was suggested that these proteins were important in regulating the uterine environment for the blastocyst implantation. By identification of DS-1-47 markers, proteomic analysis coupled with functional assays is demonstrated to be a promising approach to better understand the molecular mechanism of traditional Chinese medicine.
Animals ; Drugs, Chinese Herbal ; pharmacology ; Embryo Implantation ; drug effects ; Female ; Mice ; Ovulation Induction ; Pregnancy ; Proteome ; genetics ; metabolism ; Uterus ; drug effects ; metabolism ; physiology

OBJECTIVETo explore the protective effect of electroacupuncture (EA) pretreatment on ovarian function in rats with ovulation induction.

METHODSThirty SD female rats were numbered according to random number table. According to vaginal smear method, rats of estrus were divided into a normal group (10 rats) and cohabitated with male SD rats with the proportion of 1:1. With computer-generated random number, the remaining rats were divided into a model group and an EA group, 10 rats in each one. The model of superovulation was established with pregnant mare serum gonadotropin (PMSG) and human chorionic gonadotropin (HCG) in the model group and EA group. Before model establishment and cohabitation, rats in the EA group were treated with EA at "Guanyuan (CV 4)" and "Sanyinjiao (SP 6)", once for 15 min, for consecutive 7 days. Rats in the normal group and model group received no further treatment. The third day 23:00 pm after cohabitation, blood samples in three groups were collected to test the level of estradiol (E₂) and progesterone (P). After the rats were sacrificed, the HE staining method was applied to observe the morphological changes of ovarian tissue; the immunohistochemical method was applied to measure the expression of vascular endothelial growth factor (VEGF) and its receptor VEGFR-2; the real-time quantitative PCR technique was applied to measure the gene expression of VEGF and VEGFR-2.

RESULTSThe number of ovarian follicle in the EA group was higher than that in the model group and normal group (all P < 0.05); the ratio of corpus luteum size to ovarian size in the EA group was lower than that in the model group (P < 0.01). The ratio of plasma estradiol to progesterone in the EA group tended to be normal group (P < 0.05) and lower than that in the model group (P < 0.01). The protein expression of VEGF and VEGFR-2 in lutein granulosa cell and follicular fluid in the EA group was lower than that in the model group (P < 0.05); gene level of VEGF and VEGFR-2 in ovarian tissue in the EA group was lower than that in the model group (P < 0.05, P < 0.01).

CONCLUSIONEA pretreatment has certain protective effect on ovarian function in rats with ovulation induction, which is likely to be related to regulation of VEGF and its receptor.

Acupuncture Points ; Animals ; Chorionic Gonadotropin ; blood ; Electroacupuncture ; Estradiol ; blood ; Female ; Male ; Ovary ; physiology ; Ovulation Induction ; Pregnancy ; Progesterone ; blood ; Rats ; Rats, Sprague-Dawley ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

Serum anti-Mullerian hormone (AMH) levels are regarded as an age-specific marker for predicting the ovarian reserve in women of reproductive age. Some studies have shown that the luteinizing hormone (LH)/follicle stimulating hormone (FSH) ratio can be used as a predictor of ovarian reserve. The purpose of this study was to assess the variation of LH/FSH ratio with aging and to evaluate the correlation between serum LH/FSH ratio and AMH levels as a predictor of the ovarian reserve in normo-ovulatory women. We retrospectively analyzed the day 3 serum hormone levels in 1,251 patients (age range: 20-50 yr) between January 2010 and January 2011. We divided the patients into 6 groups according to their age. Relation between serum AMH level and LH/FSH ratio was analyzed statistically. The serum AMH level was inversely correlated with age (r = -0.400, P < 0.001). A significant negative correlation was found between serum LH/FSH ratio and age (r = -0.213, P < 0.001). There was a significant partial correlation between serum LH/FSH ratio and AMH level when adjusted by age (r = 0.348, P < 0.001). The LH/FSH ratio could be considered as a useful marker for the ovarian reserve and could be applied to the clinical evaluation with AMH.
Adult ; Aging/blood ; Anti-Mullerian Hormone/*blood ; Female ; Follicle Stimulating Hormone/*blood ; Humans ; Luteinizing Hormone/*blood ; Menstruation/blood/physiology ; Middle Aged ; Ovarian Reserve/*physiology ; Ovulation/*blood/physiology ; Retrospective Studies ; Young Adult

Serum anti-Mullerian hormone (AMH) levels are regarded as an age-specific marker for predicting the ovarian reserve in women of reproductive age. Some studies have shown that the luteinizing hormone (LH)/follicle stimulating hormone (FSH) ratio can be used as a predictor of ovarian reserve. The purpose of this study was to assess the variation of LH/FSH ratio with aging and to evaluate the correlation between serum LH/FSH ratio and AMH levels as a predictor of the ovarian reserve in normo-ovulatory women. We retrospectively analyzed the day 3 serum hormone levels in 1,251 patients (age range: 20-50 yr) between January 2010 and January 2011. We divided the patients into 6 groups according to their age. Relation between serum AMH level and LH/FSH ratio was analyzed statistically. The serum AMH level was inversely correlated with age (r = -0.400, P < 0.001). A significant negative correlation was found between serum LH/FSH ratio and age (r = -0.213, P < 0.001). There was a significant partial correlation between serum LH/FSH ratio and AMH level when adjusted by age (r = 0.348, P < 0.001). The LH/FSH ratio could be considered as a useful marker for the ovarian reserve and could be applied to the clinical evaluation with AMH.
Adult ; Aging/blood ; Anti-Mullerian Hormone/*blood ; Female ; Follicle Stimulating Hormone/*blood ; Humans ; Luteinizing Hormone/*blood ; Menstruation/blood/physiology ; Middle Aged ; Ovarian Reserve/*physiology ; Ovulation/*blood/physiology ; Retrospective Studies ; Young Adult

The objective of this study was to investigate factors that influence the success of resynchronization protocols for bovines with and without progesterone supplementation. Cow synchronized and not found pregnant were randomly assigned to two resynchronization protocols: ovsynch without progesterone (P4) supplementation (n = 66) or with exogenous P4 administered from Days 0 to 7 (n = 67). Progesterone levels were measured on Days 0 and 7 of these protocols as well as 4 and 5 days post-insemination. Progesterone supplementation raised the P4 levels on Day 7 (p < 0.05), but had no overall effect on resynchronization rates (RRs) or pregnancy per artificial insemination (P/AI). However, cows with Body Condition Score (BCS) > 3.5 had increased P/AI values while cows with BCS < 2.75 had decreased P/AI rates after P4 supplementation. Primiparous cows had higher P4 values on Day 7 than pluriparous animals (p = 0.04) and tended to have higher RRs (p = 0.06). Results of this study indicate that progesterone supplementation in resynchronization protocols has minimal effects on outcomes. Parity had an effect on the levels of circulating progesterone at initiation of the protocol, which in turn influenced the RR.
Animals ; Cattle/*physiology ; Dinoprost/administration & dosage/*pharmacology ; Estrus Synchronization/*drug effects/methods ; Female ; Fertility Agents/administration & dosage/pharmacology ; Gonadotropin-Releasing Hormone/administration & dosage/*pharmacology ; Insemination, Artificial/veterinary ; Ovulation/drug effects ; Pregnancy ; Progesterone/administration & dosage/*pharmacology ; Tromethamine/administration & dosage/*pharmacology

The aim of this study was to investigate the effect of Bu-Shen-An-Tai recipe (BSATR) and its two components (Bushen recipe, and Huoxue recipe) on endometrial morphology during peri-implantation in superovulated mice. Mice were randomly divided into five groups, including the normal (N), model (M), Bushen (BS), Huoxue (HX) and Bu-Shen-An-Tai (BH) groups. The uteri were collected on day 4 of pregnancy, and the endometrium thickness, microvessel density (MVD) and number of pinopodes observed. Compared with the M group, the endometrial thickness in the BS, HX and BH groups was significantly increased and there was a significant difference in endometrial thickness between the BS and the BH groups. The mean MVD was significantly lower in the M group than in the N group, and there was a significant increase in MVD in the BS, HX and BH groups as compared with the M group. Compared with the M group, the pinopode scores in the endometrium were significantly increased in the HX and BH groups; and the BS group had significantly higher pinipode scores than the HX and BH groups. In conclusion, the results of the present study demonstrated that the recipes (Bushen, Huoxue and BSATR) could improve the endometrial environment by regulating the endometrial thickness, MVD and the number of pinopodes at the window of implantation. Moreover, the Huoxue recipe and the BSATR were more efficient than the Bushen recipe, with the BSATR tending to have the most beneficial effects.
Animals ; Antigens, CD34 ; metabolism ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Embryo Implantation ; physiology ; Endometrium ; drug effects ; physiology ; ultrastructure ; Female ; Immunohistochemistry ; Mice ; Microscopy, Electron, Scanning ; Microvessels ; metabolism ; physiology ; Ovulation ; physiology ; Pregnancy ; Random Allocation ; Time Factors

OBJECTIVETo study the effects and underlying mechanisms of Zhuyun Recipe (ZR) on the endometrial receptivity in ovarian stimulation (OS) and blastocyst implantation dysfunction (BID) mice.

METHODSTotally 200 normal female Kunming mice were randomly divided into 6 groups, i. e., the control group (Group A), the OS group (Group B), the OS + ZR group (Group C), the BID group (Group D), the BID + ZR group (Group E), and the ZR group (Group F). The pregnant mare's serum gonadotrophin (PMSG) and human chorionic gonadotrophin (HCG) were intraperitoneally injected to mice in Group B. Mifepristone was subcutaneously injected to mice in Group D at 9:00 am on the 4th gestation day. Corresponding medications were given to mice in Group C, E, and F at 1.5 mL/100 g by gastrogavage at 8:00 am from the first to the 4th gestation day. Eight uterus samples were collected at 9:00 pm on the 4th gestation day and fixed. The expression levels of leukemia inhibitory factor (LIF) and integrin beta3 were detected using immunohistochemical assay. The pregnant mice were sacrificed at 9:30 pm on the 8th gestation day, and their uterus were taken out. The number of blastocysts was counted.

RESULTSCompared with Group A, the pregnant rate was 6.67% (1/15 cases) in Group B and 18.75% (3/16 cases) in Group D, the mean OD value of LIF was 0. 18 +/- 0.02 in Group B and 0.23 +/- 0.02 in Group D, and the mean OD value of integrin beta3 was 0.20 +/- 0.05 in Group B and 0.19 +/- 0. 02 in Group D, showing statistical difference (P < 0.01). The pregnant rate was 54.55% (12/22 cases) in Group C and 65. 22% (15/23 cases) in Group E, the mean OD value of LIF was 0.37 +/- 0. 09 in Group C and 0.39 +/- 0.02 in Group E, and the mean OD value of integrin beta3 was 0.34 +/- 0.04 in Group C and 0.38 +/- 0.08 in Group E, showing statistical difference when compared with those of Group B and Group D respectively (P < 0.05).

CONCLUSIONSOS and BID had negative effects on the endometrial receptivity and hindered the blastocyst implantation. ZR could improve the uterine receptivity and elevate the pregnant rate by up-regulating the expressions of endometrial LIF and integrin beta3.

Animals ; Drugs, Chinese Herbal ; pharmacology ; Embryo Implantation ; drug effects ; Endometrium ; drug effects ; physiology ; Female ; Integrin beta3 ; metabolism ; Leukemia Inhibitory Factor ; metabolism ; Mice ; Mice, Inbred Strains ; Ovulation Induction ; Pregnancy

OBJECTIVETo study the clinical effects of Yupei Qisun Sequential Method (YQSM, by Shen supplementing and Pi invigorating) of Chinese medicine on correlated indices of repeated implantation failure patients in the fresh cycle.

METHODSSixty patients with more than three failure cycles of in virto fertilization and embryo transfer (IVF-ET) or intracytoplasmic sperm injection (ICSI) were recruited. They were assigned to the treatment group (treated by IVF/ICSI and Chinese medicine) and the control group (treated by IVF/ICSI alone), 30 in each group. The total dose of gonadotropin (Gn), the days of controlled ovary hyperstimulation (COH), the thickness of endometrium on the day of embryo transplantation, the number of retrieved oocytes, the fertilization number, the embryo number, the high quality embryo number, the pregnancy rate, and the implantation rate were compared.

RESULTSIn the treatment group the numbers of embryo and high quality embryo were 7.5 +/- 4.9 and 5.1 +/- 3.2 respectively, which were both higher than those of the control group with significant difference (5.1 +/- 3.2, 3.2 +/- 1.8; P < 0.05). No significant difference existed in aspects of the total numbers of Gn, the days of COH, the thickness of endometrium on the day of embryo transplantation, the numbers of retrieved oocytes, the fertilization number, the pregnancy rate, and the implantation rate between the two groups (P > 0.05).

CONCLUSIONYQSM combined with COH could improve the quantity and the quality of embryos, which was promising to increase the accumulative pregnancy rate of RIF patients.

Adult ; Embryo Transfer ; Endometrium ; physiology ; Female ; Fertilization in Vitro ; Gonadotropins ; administration & dosage ; Humans ; Medicine, Chinese Traditional ; methods ; Oocyte Retrieval ; Ovulation Induction ; Pregnancy ; Pregnancy Rate ; Sperm Injections, Intracytoplasmic ; Treatment Failure