Prevalence of polycystic ovary syndrome under NIH criteria among the tenth-grade Chinese schoolgirls in Guangzhou area: a cross-sectional epidemiological survey.

BACKGROUND: Polycystic ovary syndrome (PCOS) is currently considered to have a peri-adolescence onset and continuously influence the reproductive and metabolic health of the patients, while the diagnostic criteria among adolescent population haven't been universally unified till now. This survey seeks to preliminarily evaluate the prevalence of PCOS in the tenth grade schoolgirls in Guangzhou area under NIH criteria and analyze the clinical features of adolescents with PCOS. METHODS: The cross-sectional epidemiological survey was carried out among the tenth grade schoolgirls in Guangzhou area by the method of cluster sampling. The contents of this survey included the questionnaire, physical exams and serum measurements. Until now, totally 1294 girls underwent this survey and 1095 serum samples were restored. 235 non-hirsute (mFG < 6), postmenarcheally 2-year girls were randomly selected as the control group, among which the cut-off value of biochemical hyperandrogenemia was set accordingly. The prevalence of PCOS among this population was preliminarily evaluated according to the NIH criteria. RESULTS: Along with the increase of gynecological age, the menstruations of girls was becoming more regular and the incidence of oligomenorrhea or amenorrhea was declining. Even among those who were less than 2 years after menarche, those whose menstrual cycle were longer than 90 days accounted for lower than 5%. The 95th percentile of mFG score was 6 among the girls who were < 2 years after menarche, and 5 among the girls who were > 2 years after menarche. Among the 235 healthy girls, the 95th percentile values of Testosterone (T), Free androgen index (FAI) and Androstenedione (A2) were 2.28 nmol/mL, 4.37, and 5.20 nmol/mL respectively. Based on the NIH criteria, the prevalence of PCOS in this survey was 3.86%. The prevalence of adolescent PCOS tend to slightly increase with age and gynecological age, but the difference was not statistically significant. The prevalence of PCOS among obese girls was markedly higher than that in lean girls. CONCLUSION: Based on the NIH criteria, the prevalence of PCOS among the tenth grade schoolgirls in Guangzhou area was 3.86%. The diagnosis of hyperandrogenism among adolescents should also be based on both clinical and biochemical parameters.

A Phase III randomized controlled trial of oral dydrogesterone versus intravaginal progesterone gel for luteal phase support in in vitro fertilization (Lotus II): results from the Chinese mainland subpopulation.

Lotus II, a randomized, open-label, multicenter, international study compared the efficacy and safety of oral dydrogesterone versus micronized vaginal progesterone (MVP) gel for luteal support in IVF. A prespecified subgroup analysis was performed on 239 Chinese mainland subjects from the overall study population (n = 1034), who were randomized to oral dydrogesterone 30 mg or 8% MVP gel 90 mg daily from the day of oocyte retrieval until 12 weeks of gestation. The aim was to demonstrate non-inferiority of oral dydrogesterone to MVP gel, assessed by the presence of a fetal heartbeat at 12 weeks of gestation. In the Chinese mainland subpopulation, there was a numerical difference of 9.4% in favor of oral dydrogesterone, with ongoing pregnancy rates at 12 weeks of gestation of 61.4% and 51.9% in the oral dydrogesterone and MVP gel groups, respectively (adjusted difference, 9.4%; 95% CI: -3.4 to 22.1); in the overall population, these were 38.7% and 35%, respectively (adjusted difference, 3.7%; 95% CI: -2.3 to 9.7). In both the Chinese mainland subpopulation and the overall population, dydrogesterone had similar efficacy and safety to MVP gel. With convenient oral administration, dydrogesterone has potential to transform luteal support treatment.

Oral dydrogesterone versus intravaginal micronized progesterone gel for luteal phase support in IVF: a randomized clinical trial.

STUDY QUESTION: Is oral dydrogesterone 30 mg daily non-inferior to 8% micronized vaginal progesterone (MVP) gel 90 mg daily for luteal phase support in IVF? SUMMARY ANSWER: Oral dydrogesterone demonstrated non-inferiority to MVP gel for the presence of fetal heartbeats at 12 weeks of gestation (non-inferiority margin 10%). WHAT IS KNOWN ALREADY: The standard of care for luteal phase support in IVF is the use of MVP; however, it is associated with vaginal irritation, discharge and poor patient compliance. Oral dydrogesterone may replace MVP as the standard of care if it is found to be efficacious with an acceptable safety profile. STUDY DESIGN, SIZE, DURATION: Lotus II was a randomized, open-label, multicenter, Phase III, non-inferiority study conducted at 37 IVF centers in 10 countries worldwide, from August 2015 until May 2017. In total, 1034 premenopausal women (>18 to <42 years of age) undergoing IVF were randomized 1:1 (stratified by country and age group), using an Interactive Web Response System, to receive oral dydrogesterone 30 mg or 8% MVP gel 90 mg daily. PARTICIPANTS/MATERIALS, SETTING, METHODS: Subjects received either oral dydrogesterone (n = 520) or MVP gel (n = 514) on the day of oocyte retrieval, and luteal phase support continued until 12 weeks of gestation. The primary outcome measure was the presence of fetal heartbeats at 12 weeks of gestation, as determined by transvaginal ultrasound. MAIN RESULTS AND THE ROLE OF CHANCE: Non-inferiority of oral dydrogesterone was demonstrated, with pregnancy rates in the full analysis sample (FAS) at 12 weeks of gestation of 38.7% (191/494) and 35.0% (171/489) in the oral dydrogesterone and MVP gel groups, respectively (adjusted difference, 3.7%; 95% CI: -2.3 to 9.7). Live birth rates in the FAS of 34.4% (170/494) and 32.5% (159/489) were obtained for the oral dydrogesterone and MVP gel groups, respectively (adjusted difference 1.9%; 95% CI: -4.0 to 7.8). Oral dydrogesterone was well tolerated and had a similar safety profile to MVP gel. LIMITATIONS, REASONS FOR CAUTION: The analysis of the results was powered to consider the ongoing pregnancy rate, but a primary objective of greater clinical interest may have been the live birth rate. This study was open-label as it was not technically feasible to make a placebo applicator for MVP gel, which may have increased the risk of bias for the subjective endpoints reported in this study. While the use of oral dydrogesterone in fresh-cycle IVF was investigated in this study, further research is needed to investigate its efficacy in programmed frozen-thawed cycles where corpora lutea do not exist. WIDER IMPLICATIONS OF THE FINDINGS: This study demonstrates that oral dydrogesterone is a viable alternative to MVP gel, due to its comparable efficacy and tolerability profiles. Owing to its patient-friendly oral administration route, dydrogesterone may replace MVP as the standard of care for luteal phase support in fresh-cycle IVF. STUDY FUNDING/COMPETING INTERESTS(S): This study was sponsored and supported by Abbott. G.G. has received investigator fees from Abbott during the conduct of the study. Outside of this submitted work, G.G. has received non-financial support from MSD, Ferring, Merck-Serono, IBSA, Finox, TEVA, Glycotope and Gedeon Richter, as well as personal fees from MSD, Ferring, Merck-Serono, IBSA, Finox, TEVA, Glycotope, VitroLife, NMC Healthcare, ReprodWissen, Biosilu, Gedeon Richter and ZIVA. C.B. is the President of the Belgian Society of Reproductive Medicine (unpaid) and Section Editor of Reproductive BioMedicine Online. C.B. has received grants from Ferring Pharmaceuticals, participated in an MSD sponsored trial, and has received payment from Ferring, MSD, Biomérieux, Abbott and Merck for lectures. G.S. has no conflicts of interest to be declared. A.P. is the General Secretary of the Indian Society of Assisted Reproduction (2017-2018). B.D. is President of Pune Obstetric and Gynecological Society (2017-2018). D.-Z.Y. has no conflicts of interest to be declared. Z.-J.C. has no conflicts of interest to be declared. E.K. is an employee of Abbott Laboratories GmbH, Hannover, Germany and owns shares in Abbott. C.P.-F. is an employee of Abbott GmbH & Co. KG, Wiesbaden, Germany and owns shares in Abbott. H.T.'s institution has received grants from Merck, MSD, Goodlife, Cook, Roche, Origio, Besins, Ferring and Mithra (now Allergan); and H.T. has received consultancy fees from Finox-Gedeon Richter, Merck, Ferring, Abbott and ObsEva. TRIAL REGISTRATION NUMBER: NCT02491437 (clinicaltrials.gov). TRIAL REGISTRATION DATE: 08 July 2015. DATE OF FIRST PATIENT'S ENROLLMENT: 17 August 2015.

UHPLC/Q-TOFMS-based plasma metabolomics of polycystic ovary syndrome patients with and without insulin resistance.

Polycystic ovary syndrome (PCOS), characterized with menstrual irregularities, hyperandrogenism and ovulatory abnormalities, is usually companied with insulin resistance (IR) and accounts for one of the most prevalent reproductive dysfunction of premenopausal women. Despite accumulating investigations, diagnostic standards of this pathological condition remain obscure. The aim of present study is to characterize the plasma metabolic characteristics of PCOS patients with and without IR, and subsequently identify the potential biomarkers for the diagnosis of PCOS and its IR complication. A total of 59 plasma samples from eligible healthy controls (CON, n=19), PCOS patients without IR (non-IR PCOS, n=19) and PCOS patients with IR (IR PCOS, n=21) were profiled by an ultra high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC/Q-TOFMS) followed by multivariate statistical analysis. Compared to the healthy controls, significant decrease in the levels of phosphocholines (PCs) and lyso PC (18:2), and increase in trilauric glyceride level were observed in the plasma of IR PCOS. Meanwhile, the significant increase in the levels of saturated fatty acids (palmitic acid and stearic acid) and decanoylcarnitine, and decrease in PC (36:2) and PS (36:0) were found in non-IR PCOS patients. Trilauric glyceride and decanoylcarnitine were identified as the potential biomarkers with the highest sensitivity and specificity for the diagnosis of PCOS patients with and without IR, respectively. Furthermore, based on these alterations of metabolites, MetPA network pathway analysis suggested a profound involvement of the abnormalities of glycerophospholipid, glycerolipid and fatty acid metabolisms in the pathogenesis of PCOS and IR complications. Collectively, LC-MS-based metabolomics provides a promising strategy for complementary diagnosis of PCOS and its IR complication and offers a new insight to understand their pathogenesis mechanisms.

Does reducing gamete co-incubation time improve clinical outcomes: a retrospective study.

PURPOSE: The objective of this retrospective study was to determine whether patients undergoing in vitro fertilization (IVF) benefit from reducing the gamete co-incubation time. METHODS: Patients (n = 570) were enrolled, including 281 patients in the reduced incubation time group (2-h incubation) and 289 patients in the standard IVF group (18-h incubation). RESULTS: The observed outcomes, including the clinical pregnancy rate (CPR), implantation rate (IR), live birth rate (LBR), and miscarriage rate (MR), were similar between the two groups. When the data were divided into two subgroups based on the maternal age (≤30 and >30 years), the rates of top-quality embryos (30.83 vs. 25.89 %; p = 0.028), CPR (66.67 vs. 42.11 %; p = 0.013), and IR (41.90 vs. 31.25 %, p = 0.019) of the 2-h incubation group were significantly higher in the younger subgroup. However, for older patients, only a lower MR (7.59 vs. 20.83 %; p = 0.019) was achieved. Reducing the time of incubation still improved the CPR (OR = 1.993, 95 % CI 1.141-3.480) and MR (OR = 3.173, 95 % CI 1.013-9.936) in the younger and older subgroups, respectively, after it was adjusted for potential confounders. CONCLUSIONS: Reducing incubation time improves the clinical results of IVF, although the LBR is not statistically different between the 2- and 18-h incubation time groups. And the specific clinical outcomes of reducing incubation time varied between the >30-year-old and the ≤30-year-old.

Identification of differentially expressed microRNAs in the ovary of polycystic ovary syndrome with hyperandrogenism and insulin resistance.

BACKGROUND: Polycystic ovary syndrome (PCOS) is the commonest endocrinopathy in women of reproductive age. The patients often develop insulin resistance (IR) or hyperinsulinemia despite manifesting anovulation and signs of hyperandrogenism. The cause and effect relationship of hyperinsulinemia and hyperandrogenemia (HA) is still debated. Micro-ribonucleic acids (miRNAs) have recently been shown to play a role in regulation of ovarian function. Our current study focused on the altered expression of miRNAs with PCOS. METHODS: Ovarian theca interna tissues were obtained from 10 PCOS patients and 8 controls that were non-PCOS and had normal insulin sensitivity undergoing laparoscopy and/or ovarian wedge resection. Total RNA of all samples was extracted. We studied the repertoire of miRNAs in both PCOS and non-PCOS women by microarray hybridization. Bioinformatic analysis was performed for predicting targets of the differentially expressed miRNAs. Furthermore, selected miRNAs were validated by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR). RESULTS: A total of 27 miRNAs were differentially expressed in PCOS patients with respect to the controls in our discovery evaluationand two (miR-92a and miR-92b) of them were significantly downregulated in PCOS women in followed validation (P < 0.05). Targets prediction revealed that miR-92a targeted both GATA family of zinc finger transcription factor GATA-binding factor 6 (GATA6) and insulin receptor substrate proteins 2 (IRS-2). CONCLUSIONS: MiRNAs are differentially expressed between PCOS patients and controls. We identified and validated two miRNAs-miR-92a and miR-92b. They are significantly downregulated and may be involved in the pathogenesis of PCOS.

Association between the proportion of dominant follicles and oocyte developmental competence.

PURPOSE: To explore the optimal timing for hCG triggering by investigating the impact of different proportion of dominant follicles on the oocyte developmental competence. METHODS: One hundred ninety-eight infertile women were divided into three groups according to the proportion of dominant follicles on hCG day: (1) low: <15% (n = 66); (2) middle: 15-27% (n = 66); (3) high: >27% (n = 66). The grouping criteria were the bottom and top tertiles of the proportion of dominant follicles. RESULTS: The gonadotropin dosage, duration and maximum follicle diameter in the low proportion group were lower than those in the middle and high proportion groups. Oocyte maturation and the abnormal fertilization rate in the low proportion group were lower than those in the middle and high proportion groups. The normal fertilization rate did not differ among the three groups. The cleavage rate and number of transferable embryos in the low proportion group were significantly higher than those in the high proportion group. The high-quality embryo rate, implantation rate, and pregnancy rate in the low proportion group were significantly higher than those in the middle and high proportion groups. CONCLUSIONS: A high proportion of dominant follicles are closely associated with impaired oocyte developmental competence and low pregnancy rate. These findings suggest that follicular overgrowth induced by delayed hCG triggering may undermine oocyte developmental competence and the proportion of dominant follicles may be a potential parameters for hCG triggering.

Increased GDF9 and BMP15 mRNA levels in cumulus granulosa cells correlate with oocyte maturation, fertilization, and embryo quality in humans.

BACKGROUND: Oocyte secreted factors (OSFs), including growth differentiation factor 9 (GDF9) and bone morphogenetic protein 15 (BMP15), play an important role in the process of follicular development and oocyte maturation. Since OSFs are expressed in oocytes and cumulus granulosa cells, the aim of the present study was to explore whether the expression levels of GDF9 and BMP15 mRNAs in cumulus granulosa cells can be used as molecular markers for predicting oocyte developmental potential. METHODS: Cumulus cells of 2426 cumulus-oocyte complexes were collected from 196 female patients who underwent intracytoplasmic sperm injection (ICSI) and were used for mRNA detection on the egg retrieval day. Pearson correlation analysis was used to analyze the correlation between OSF expression and general physiological parameters. Partial correlation analysis was used to analyze the correlation between OSF expression and oocyte developmental potential. Covariance analysis was used to compare OSF expression among different groups. Receiver operating characteristic curves were used to examine the diagnostic value of GDF9 and BMP15 mRNA for predicting pregnancy. RESULTS: The expression levels of GDF9 and BMP15 mRNAs were significantly associated with age, body mass index (BMI), oocyte maturation, normal fertilization, and cleavage rate (P < 0.05). The expression levels of GDF9 and BMP15 mRNAs in the group with high-quality embryos were significantly higher than those in the group without high-quality embryos (P < 0.05). The expression levels of GDF9 and BMP15 mRNAs in the pregnancy group were significantly higher than those in the nonpregnancy group (P < 0.05). The cut-off value of GDF9 mRNA for predicting pregnancy was 4.82, with a sensitivity of 82% and a specificity of 64%. The cut-off value of BMP15 mRNA for predicting pregnancy was 2.60, with a sensitivity of 78% and a specificity of 52%. CONCLUSIONS: The expression levels of GDF9 and BMP15 mRNAs were closely associated with oocyte maturation, fertilization, embryo quality, and pregnancy outcome; therefore, GDF9 and BMP15 mRNAs in cumulus granulosa cells may be considered as new molecular markers for predicting oocyte developmental potential.

Characterizing plasma phospholipid fatty acid profiles of polycystic ovary syndrome patients with and without insulin resistance using GC-MS and chemometrics approach.

Polycystic ovary syndrome (PCOS), a heterogeneous endocrine and metabolic disorder, is the leading cause of infertility in women of reproductive age. Insulin resistance (IR) occurs in 50-70% of women with PCOS. In this study, we aimed to characterize the plasma phospholipid fatty acid profile for PCOS patients with and without IR, as well as for the early prognosis of PCOS and its IR complication. A gas chromatography-mass spectrometry (GC-MS) followed by multivariate statistical analysis was established to globally characterize the phospholipid fatty acid profiles in plasma from non-IR PCOS, IR PCOS, and eligible healthy controls, and subsequently discovered fatty acid biomarkers. A total of 22 fatty acids were identified and quantified. Their proportions varied among three groups, suggesting each group has its own fatty acid pattern. Orthogonal partial least squares discriminant analysis (OPLS-DA) according to their fatty acid profiles showed that 29 tested samples could be clearly differentiated according to groups. More importantly, nervonic acid (C24:1 n-9) and dihomo-γ-linolenic acid (C20:3 n-6) were identified as the potential fatty acid biomarkers of PCOS and its IR complication, respectively, for their most contribution to group separation. Pearson correlation analysis indicated that C24:1 n-9 and C20:3 n-6 were well correlated with clinical characteristics of PCOS and IR indicators, respectively. These findings demonstrated that GC-MS-based plasma phospholipid fatty acid profile might provide a complementary approach for clinical diagnosis of PCOS and its IR complication.

[Study on the facial and body terminal hair growth in women in Guangdong by using modified Ferriman-Gallwey scoring system].

OBJECTIVE: To study growth of facial and body terminal hair of women in Guangdong province and its relationship with age, menstrual irregularities and polycystic ovary, and determine normative cut-off score of modified Ferriman and Gallwey (mFG). METHODS: A cross-sectional study was conducted on 2988 women at age of 20-45 years from 16 communities of two urban and two rural regions in Guangdong province from June 2008 to July 2009. Terminal body hair growth was assessed by using the modified Ferriman and Gallwey (mFG) scoring system. The normative cut-off value of mFG were calculated by using the K-means cluster analysis (K=2). Those women were classified into following groups, including 982 women at group of ages of 20- years, 765 women at group of 26- years, 597 women at group of 31- years, 384 women at group of 36- years, 260 women at group of 41-45 years. Due to absence or errors of medical records, some cases were excluded from this study. Based on menses irregularities (MI), polycystic ovaries (PCO), there were 488 cases in MI group, 2413 cases in normal menses group, 568 cases in PCO group, and 2207 cases in non-PCO group finally. The incidences of acne, MI, acanthosis nigricans, and polycystic ovaries were also analyzed in all the hirsute groups. RESULTS: (1) among 2988 women, it was observed 149 women (5%) with mFG≥7,314 women (10.5%) with ≥5,747 women with mFG≥2. (2) Cluster analysis identified an mFG score of 5 as the cut-off value that define abnormal hirsute in the total population and all the sub-groups with/without MI or PCO; (3) Based on age classification, it was found that increased age was associated with decreased trends of the percentile and cut-off value of hirsutism. The value of hirsutism of mFG were 6 in group of 20- years, 5 in group of 26- years, 4 in groups of 31- years, 36- years and 41-45 years. (4) The prevalence of acne, menstrual irregularities and POC were 45.5% (143/314), 73.6% (231/314), 25.8% (81/314) in total population, 25.1% (671/2674), 16.1% (431/2674), 19.8% (529/2674) in normal hair women, which reached statistical difference (P<0.05). The prevalence of acne, menstrual irregularities and acanthosis nigricans were 44.4% (130/293), 23.2% (68/293), 4.1% (12/293) in those age hirsute groups, 25.3% (681/2695), 16.2% (437/2695), 1.9% (51/2695) in normal hair women, which reached statistical difference (P<0.05). CONCLUSIONS: (1) among women in Guangdong province, mFG scoring showed decreased trends in women with increasing age. (2) An mFG score≥5 was cut-off value in diagnosis of hirsutism. (3) The hirsute women exhibited higher incidence of acne, menses irregularity, and acanthosis nigricans than those of women with normal hair growth.

Progress in understanding human ovarian folliculogenesis and its implications in assisted reproduction.

PURPOSE: To highlight recent progress in understanding the pattern of follicular wave emergence of human menstrual cycle, providing a brief overview of the new options for human ovarian stimulation and oocyte retrieval by making full use of follicular physiological waves of the patients either with normal or abnormal ovarian reserve. METHODS: Literature review and editorial commentary. RESULTS: There has been increasing evidence to suggest that multiple (two or three) antral follicular waves are recruited during human menstrual cycle. The treatment regimens designed based on the theory of follicular waves, to promote increased success with assisted reproduction technology (ART) and fertility preservation have been reported. These new options for human ovarian stimulation and oocyte retrieval by making full use of follicular waves of the patients either with normal or abnormal ovarian reserve lead to new thinking about the standard protocols in ART and challenge the traditional theory that a single wave of antral follicles grows only during the follicular phase of the menstrual cycle. CONCLUSIONS: The understanding of human ovarian folliculogenesis may have profound implications in ART and fertility preservation. Further studies are needed to evaluate the optimal regimens in ART based on the theory of follicular waves and to identify non-invasive markers for predicting the outcome and the potential utilities of follicles obtained from anovulatory follicular waves in ART.

Protection of CHO cells by transfer of survivin driven by ovarian-specific promoter OSP-2.

Chemotherapy is the major therapy for cancer in clinic. However, chemotherapeutic agents can harm the other tissues/organs besides cancer. Thus, there are great interests in protecting the innocents by the transfer of protective genes. There are two problems to be solved, one is the selection of protective genes and the other is the orientation of the exotic genes. Recent researches demonstrated that the principal mechanism of chemotherapeutics was through apoptosis. Hereby, introduction of anti-apoptosis genes might interrupt the processes of apoptosis to avoid side effect from chemotherapeutics. On the other hand, tissue-specific promoters, which control gene expression in a tissue-specific manner, might be an alternative tool to guarantee the location of target genes. In this research, we applied gene therapy to chemoprotection using anti-apoptosis gene survivin and ovarian-specific promoter OSP-2. The results showed that OSP-2 could specifically drive the expression of survivin in ovarian cells and survivin could protect cells via inhibiting apoptosis. This might put a light on the future of chemoprotective gene therapy.

[Effects of gonadotroph-releasing hormone analogues on follicle apoptosis in rats with chemotherapy-induced ovarian damage].

OBJECTIVE: To study the effects of gonadotroph-releasing hormone (GnRH) agonist (GnRH-a) and GnRH antagonist (GnRH-ant) on cyclophosphamide (CTX)-induced follicle apoptosis in female rats. METHODS: Thirty-six female Sprague- Dawley rats were randomized into 6 groups, namely normal saline (NS), CTX, GnRH-a+NS, GnRH-a+CTX, GnRH-ant+NS, and GnRH-ant+CTX groups. The rats were sacrificed between the first and second week after the treatments., and the follicle apoptosis was investigated using TUNEL assay and transmission electron microscopy. RESULTS: The apoptosis rate of the granulose cells in the follicles in late development was significantly higher than that in early follicles, and the apoptosis rate of the oocytes and granulose cells in rats with CTX treatment was significantly higher than that in rats without CTX treatment (P<0.05). The apoptosis rate of the granulose cells in GnRH-a groups (ranging from 33.40 - or + 4.59 to 73.25 - or + 5.35) was significantly higher than that in GnRH-ant groups (27.46 - or + 4.52 to 49.38 - or + 5.02, P<0.05), but there was no significant difference in the oocytes of early follicles between GnRH-a groups (23.48 - or + 4.25 to 36.15 - or + 4.23) and GnRH-ant groups (21.47 - or + 3.81 to 34.04 - or + 5.54, P>0.05). Electron microscopy revealed characteristic apoptotic changes of the oocytes in early follicles and granulose cells in early and late follicles. The apoptotic changes were especially typical in the granulose cells showing the formation of the apoptotic bodies, and the oocytes only showed chromatin condensation and aggregation. CONCLUSION: In the rat mode, GnRH-a promotes while GnRH-ant suppressed follicle apoptosis induced by CTX. GnRH analogues regulates mainly granulose cell apoptosis, but have little effect on oocyte apoptosis.

[Effect of anti-Müllerian hormone on P450 aromatase mRNA expression in cultured human luteinized granulose cells].

OBJECTIVE: To investigate the effect of anti-Müllerian hormone (AMH) on hormone secretion and P450 aromatase mRNA expression from cultured human luteinized granulosa cells. METHODS: Human luteinized granulose cells were derived from 10 patients treated by in vitro fertilization-embryo transplantation (IVF-ET) in the Second Affiliated Hospital of Sun Yat-sen University from June to December 2006. Granulose cells were divided into group A, B, C, D, E depending on different concentration of AMH, testosterone group and blank group. 1x10(-7) mol/L testosterone and 1, 5, 10, 20, 50 microg/L AMH were added into the culture medium of group A, B, C, D and E. 1x10(-7) mol/L testosterone was added into the culture medium of testosterone group while no other ingredient was added into the medium of blank group. Estrogen levels in supernates were measured at 24, 48, 72 hours after cell incubation. RT-PCR was performed to detect the P450 aromatase mRNA expression in group B, C, D, E and testosterone group at 72 hours after cell incubation. RESULTS: (1) Estrogen levels in supernates of granulose cell culture at 24, 48, 72 hours were (8.529+/-0.381)x10(4), (10.977+/-0.436)x10(4), (13.309+/-0.506)x10(4) pmol/L in group A, (7.027+/-0.276)x10(4), (9.167+/-0.300)x10(4), (10.794+/-0.555)x10(4) pmol/L in group B, (6.039+/-0.226)x10(4), (7.585+/-0.548)x10(4), (8.797+/-0.518)x10(4) pmol/L in group C, (5.118+/-0.460)x10(4), (5.716+/-0.496)x10(4), (6.205+/-0.667)x10(4) pmol/L in group D, (4.932+/-0.148)x10(4), (5.323+/-0.184)x10(4), (5.629+/-0.212)x10(4) pmol/L in group E. When compared with blank group [(0.001+/-0.001)x10(4), (0.006+/-0.003)x10(4), (0.029+/-0.011)x10(4) pmol/L], the statistical differences were observed in group A, B, C, D, E (P<0.01); when compared with testosterone group [(8.418+/-0.569)x10(4), (10.841+/-0.689)x10(4), (13.301+/-0.637)x10(4) pmol/L], the statistical differences were observed in group B, C, D and E (P<0.01); statistical differences were also observed in group C, D and E when compared with group B, and also group D and E when compared with group C (P<0.01). No significant difference was observed between group D and E (P>0.05). In group A, B, C, D, E and testosterone group, the estrogen levels at 24 hours after cell culture were significantly lower than those at 48 and 72 hours (P<0.01); statistical difference was observed between estrogen levels at 48 and 72 hours (P<0.01). No significant difference was observed among 24, 48 and 72 hours in blank group (P>0.05). (2) Relative ratios of intensity of P450 aromatase/beta-actin at 72 hours of cell culture in group B, C, D and E were 0.6148+/-0.0046, 0.5156+/-0.0012, 0.4698+/-0.0027 and 0.4282+/-0.0017, respectively, which were statistically lower than that in testosterone group (0.8224+/-0.0021, P<0.01); statistical differences were also observed in group C, D and E when compared with group B, and also group D and E when compared with group C (P<0.01). No significant difference was observed between group D and E (P>0.05). CONCLUSION: It is suggested that AMH might affect estrogen synthesis by inhibiting P450 aromatose activity so that lead to hyperandrogenism microenvironment in local ovary.

Cloning and transcriptional activity of a novel ovarian-specific promoter from rat retrovirus-like elements.

The long terminal repeats (LTRs) are the control centers for retrovirus gene expression, which possess all of the requisite signals. It has been proved that the LTRs of Moloney murine leukemia virus (MoMLV) could constitutively activate genes in diverse cell types. Recently, a retrovirus-like element, OSP-1 (ovarian-specific promoter 1), was extracted from rat ovary according to the LTRs of MoMLV, whose name was derived from the fact of ovarian-specific transcription. It was reasonable to speculate that the tissue-specificity was acquired through mutations and that there should be abound other mutants, active or inactive. In the present study, we isolated several homologous sequences to OSP-1 and detected their function. Consequently, one of them could also drive target gene expression specifically to ovarian cell lines and was named OSP-2 which shared 98% similarity to OSP-1. On the other hand, we picked up other two closest sequences and proved them inactive, which was 97% and 95% similar to OSP-1, respectively. Sequence analysis revealed the different mutations around/within the binding sites of transcriptional factors that might play important roles in tissue-specificity. In summary, we extracted a novel ovarian-specific promoter as well as other nonfunctional mutants, which in part shed light on the study of ovarian-specific transcription. In addition, it also provided a new tool in cancer gene therapy and to create transgenic animals.

[Plasma metastin in adolescent polycystic ovary syndrome.].

OBJECTIVE: This study was designed to investigate the correlationship between plasma metastin and pathogenesis of adolescent women with polycystic ovary syndrome (PCOS). METHODS: From Jan. 2006 to Jun. 2006, 42 PCOS patients including 19 adolescent women and 23 adults with syndrome were treated in Second Affiliated Hospital of Sun Yat-Sen University. According to the range of age, those patients were divided into 19 cases in adolescent group ( 19 years). Meanwhile, 20 adolescent women were matched as controls. Blood samples were collected between day 1 and day 5 of a spontaneous bleeding episode in the PCOS groups and a menstrual cycle of the controls. The levels of luteinizing hormone (LH), follicle-stimulating hormone (FSH), testosterone (T), free T (FT), dehydroepiandrosterone sulfate (DHEAS), sex hormone-binding globulin (SHBG), insulin, glucose, and metastin were detected from day 1 to day 5 of spontaneous bleeding or withdrawal bleeding by progesterone. On the next day, oral glucose tolerance test (75 g) and insulin release test were performed on those above patients and controls. The area under curve (AUC), the ratio of fasting blood glucose to insulin (GIR) and homeostasis model assessment insulin resistance index (HOMA-IR) were calculated. RESULTS: (1) The level of hormone: the level of LH was in (12 +/- 7) U/L in adult group and (12 +/- 8) U/L in adolescent PCOS group, which were significantly higher than (6 +/- 4) U/L in controls (P < 0.05). The level of FT was (2.3 +/- 1.2) pmol/L in adult group, which was significantly higher than (1.3 +/- 0.8) pmol/L in adolescent group and (1.1 +/- 0.5) pmol/L in control group (P < 0.05). It was observed that the level of (3.1 +/- 2.7)micromol/L in adolescent group was significantly lower than (6.3 +/- 2.7) micromol/L in control group (P < 0.05). Meanwhile, the level of FAI of 5.6 +/- 4.1 in adult group was significantly higher than 3.0 +/- 1.3 in control group (P < 0.05). No significant difference in FSH, T and SHBG levels among three groups were observed (P > 0.05). (2) Metastin and metabolism: Both the levels of fasting blood insulin, 2-hour insulin and AUC of insulin were (13 +/- 7) mU/L, (88 +/- 59) mU/L and (133 +/- 80) mUxL(-1)xmin(-1) in adolescent group, which were significantly higher than (7 +/- 3) mU/L, (57 +/- 29) mU/L and (82 +/- 34) mUxL(-1)xmin(-1) in control group. The fasting blood insulin of (13 +/- 7) mU/L in adolescent group was significantly higher than (9 +/- 5) mU/L in adult group. The level of fasting blood glucose and 2-hour glucose were (5.01 +/- 0.44) mmol/L and (6.48 +/- 1.16) mmol/L in adult group, which were significantly higher than (4.68 +/- 0.29) mmol/L and (5.44 +/- 0.83) mmol/L in control group and (4.67 +/- 0.30) mmol/L and (5.93 +/- 1.44) mmol/L in adolescent group. The glucose AUC of (9.99 +/- 1.85) mmolxL(-1)xmin(-1) in adult group was significantly higher than (8.42 +/- 1.53) mmolxL(-1)xmin(-1) in control group (P < 0.05). HOMA-IR of 2.6 +/- 2.0 in adolescent group was significantly higher than 1.4 +/- 0.7 in control group. GIR of 10 +/- 8 in adolescent group was significantly lower than 16 +/- 10 in control group (P < 0.05). The metastin level of (0.25 +/- 0.19) pmol/L in adolescent group and (0.29 +/- 0.29) pmol/L in adult group were all significantly higher than (0.18 +/- 0.23) pmol/L in control group (PPh glucose were observed (r = 0.256, 0.286 and 0.267. P = 0.044, 0.025 and 0.043). CONCLUSIONS: The expression of metastin in adolescent PCOS women was significantly higher that of normal adolescent women. The increased level of metastin might be associated with pathogenesis of adolescent women with PCOS.

[Effect of GnRH analogs on the expression of Bcl-2 gene family in the ovary of rats with cyclophosphamide-induced ovarian damage].

OBJECTIVE: To study the effect of gonadotroph-releasing hormone (GnRH) agonist (GnRH-a) and GnRH antagonist (GnRH-ant) against cyclophosphamide (CTX)-induced gonadotoxicity in female rats. METHODS: Thirty-six female SD rats were divided randomly into 6 groups to receive treatment with normal saline (NS), CTX, GnRH-a+NS, GnRH-a+CTX, GnRH-ant+NS, GnRH-ant+CTX, respectively. The rats were sacrificed between the first and second week after termination of the medication to compare the weight of the ovaries, the number of the primordial follicles and the follicle growth. The expressions of bcl-2 and bax mRNA in the ovaries were examined using RT-PCR. RESULTS: The number of the primordial follicles was significantly greater and that of the growing follicles significantly lower in GnRH-a+NS and GnRH-a+CTX groups than in the GnRH-ant+CTX and CTX groups (P<0.05). The rats in GnRH-a+NS and GnRH-a+CTX groups had the lowest ovarian weight among 6 the groups (P<0.05). The bcl-2 mRNA level in the GnRH-ant+NS group was significantly higher than that in the other groups (P<0.05). The Bax mRNA in the GnRH-a+NS and GnRH-a+CTX groups was significantly higher than that in the NS group (P<0.05), but close to that in the CTX group (P>0.05); bax mRNA expression in the GnRH-ant+NS group was significantly lower than that in the NS group (P<0.05), but in GnRH-ant+CTX group, its expression was close to that in the NS group (P>0.05). CONCLUSIONS: In female rats exposed to CTX, the GnRH analogs provides ovarian protection against CTX-induced gonadotoxicity by regulating the expression of the Bax mRNA in the ovary. GnRH-a may decrease the sensitivity of the follicles to CTX-induced gonadotoxicity by promoting follicle apoptosis and inhibiting follicle proliferation, and GnRH-ant increases the sensitivity to the CTX through a reverse effect on the follicles.