Women with high telomerase activity in luteinised granulosa cells have a higher pregnancy rate during in vitro fertilisation treatment.

OBJECTIVE: To study the effect of telomerase activity (TA) in human luteinised granulosa cells (GCs) on the outcome of in vitro fertilisation treatment. METHODS: Fifty-six women, aged 23 to 39 years, were enrolled and divided into four groups according to their levels of TA. RESULTS: Seventeen cases in group A exhibited nondetectable TA, 16 cases in group B expressed low levels of TA (between 0.1 and 0.65 OD × mm), 14 cases in group C expressed moderate TA levels (between 0.66 and 1.00 OD × mm) and 9 cases in group D expressed high levels of TA (more than 1.00 OD × mm). The level of total serum testosterone (T) was significantly higher in groups C and D than in group A (1.43±0.10 vs. 1.08±0.11 nmol/L, P<0.030 and 1.56±0.08 vs. 1.08±0.11 nmol/L, P<0.005, respectively). The TA level was positively correlated with T (r=0.291, P<0.011). No obvious differences were observed in rates of fertilisation, cleavage, mature oocyte formation or good-quality embryos among the groups. The patients in group D exhibited the highest rates of embryo implantation and clinical pregnancy (with rates of 52.63% and 77.78%, respectively, compared with 18.92% and 29.41% in group A, 25.71% and 37.50% in group B and 48% and 50% in group C, with P<0.018 and P=0.112, respectively). The patients in group D also had a greater likelihood of becoming pregnant than those in group A (OR: 9.703, P < 0.023), group B (OR: 14.765, P<0.009) or group C (OR: 5.560, P=0.103). CONCLUSIONS: Luteinised GCs have a certain potential for proliferation and TA of luteinised GCs may predict the clinical outcomes of IVF treatment. Some unknown regulatory mechanisms between TA and T should be studied in further trials.

Defining hirsutism in Chinese women: a cross-sectional study.

OBJECTIVE: To determine the normative cut-off score that defines hirsutism among Chinese women, and the effect of age, menstrual irregularities, and polycystic ovaries on the same. DESIGN: Cross-sectional, population-based study. SETTING: A multistage systematic cluster random sampling among 16 communities from two rural and two city regions. PATIENT(S): A representative sample of 2,988 women aged 20-45 years from the general population of Southern China. INTERVENTION(S): Subjects underwent physical and ultrasound evaluations. MAIN OUTCOME MEASURE(S): Terminal hair growth was assessed using the modified Ferriman-Gallwey (mFG) scoring system. RESULT(S): An mFG score ≥5 was observed in 10% and a score of ≥2 in 25% of the subjects. Cluster analysis identified an mFG score of 5 as the cut-off value that defines abnormal terminal facial and body hair growth in a male pattern (i.e., hirsutism) in the total population; scores of 6, 5, and 4 for women aged 20-25, 26-30, and >30 years, respectively. Defined by these cut-off values, the prevalence of hirsutism in our total population was 10.5%; and decreased with increasing age: 14.4%, 10.7%, 7.9%, 3.6%, and 1.5%, respectively, in women aged 20-25, 26-30, 31-35, 36-40, and 41-45 years. Furthermore, the incidence of acne, menses irregularities, polycystic ovaries, and acanthosis nigricans were significantly increased among the hirsute women. CONCLUSION(S): An mFG score of 5 or greater indicates hair growth above the norm among women in the general Southern Chinese population, a cut-off value that decreases with increasing age.

Regulation of AMH and SCF expression in human granulosa cells by GnRH agonist and antagonist.

With the progress of cancer treatment, fertility preservation has become an urgent requisition. Gonadotropin-releasing hormone agonist (GnRH-a) and antagonist (GnRH-ant) have been used to protect fertility for these patients. However, studies showed that although GnRH-a and GnRH-ant had a comparable down-regulating effect on the pituitary, GnRH-ant could not preserve ovarian function. Moreover, GnRH-ant alone could deplete primordial follicles. It might be speculated that an additional intraovarian system existed except the pituitary system. Anti-Müerian hormone (AMH) and Stem cell factor (SCF) proved to be the key factors in follicle recruitment and development. The balance between AMH and SCF was tightly related to ovarian reserve. To investigate the intraovarian effect of GnRH-a or GnRH-ant on ovarian reserve, we examined AMH and SCF expression in human granulosa cells (hGCs). GCs were isolated from follicular aspirates after oocyte removal from the patients undergoing assisted reproduction techniques. After pretreated with GnRH-a (triptorelin) or GnRH-ant (cetrorelix) for 48 h, mRNA and protein of AMH and SCF were analyzed by Real-time PCR and Immunoblot assay respectively. The results indicated that AMH mRNA and protein expressions were down-regulated in the GnRH-ant groups, SCF mRNA and protein expressions were up-regulated in the high-dose GnRH-ant group. There was no difference of AMH and SCF expression in the GnRH-a group or GnRH-a + GnRH-ant group compared with control. These results suggested the effects of GnRH-a and GnRH-ant on the regulation of AMH and SCF were different, which may provide insight into the mechanism of GnRH-a and GnRH-ant interventions on ovarian reserve.

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.

Determining the normal cut-off levels for hyperandrogenemia in Chinese women of reproductive age.

OBJECTIVE: To determine the normative cut-off levels of free testosterone (FT), dehydroepiandrosterone sulfate (DHEAS) and total testosterone (TT) among reproductive age women in China. STUDY DESIGN: A total of 450 reference subjects without known factors affecting androgen levels were selected from a total study population of 904 presumably healthy women undergoing annual check-ups. The upper limits of normal levels of biochemical androgens were computed by k-means cluster analysis, with the results categorized by age and expressed as both concentrations and percentiles. RESULTS: The upper limits (cut-off levels) of normal FT, DHEAS and TT levels as determined by k-means cluster analysis are 26 pmol/L, 4.92 µmol/L and 2.39 nmol/L, respectively, in the selected reference subjects. The corresponding percentiles of the cut-off levels of FT (91.8th vs. 87.9th), DHEAS (69.8th vs. 67.6th) and TT (90.0 th vs. 83.8th) were all higher in the reference subjects than in the total study population, suggesting that there were fewer abnormal subjects with elevated androgens in the healthier reference population than in the total study population. The level of DHEAS significantly declined by age (P < 0.05), whereas there was no significant difference in the mean FT or TT levels between age groups among these women of reproductive age. CONCLUSIONS: The normal hyperandrogenism cut-off values for FT, DHEAS and TT are 26.00 pmol/L, 4.92 µmol/L and 2.39 nmol/L, respectively, among women of reproductive age in China who are without factors that might affect androgen levels. The cut-off levels in percentiles are 91.8th for FT, 69.8th for DHEAS and 90.0 th for TT.

Metabolic abnormalities in adolescents with polycystic ovary syndrome in south China.

BACKGROUND: Adults with polycystic ovary syndrome (PCOS) can have multiple metabolic abnormalities. However, studies in the adolescent population are still limited and these results seem to vary widely. This study was to investigate the metabolic abnormalities in adolescents with PCOS in South China and the potential risk factors contributed to these health risks. METHODS: Anthropometric measurements and biochemical parameters were evaluated in 128 adolescents with PCOS and their age- and BMI-matched controls. RESULTS: The prevalence of pre-diabetes, insulin resistance, hyperinsulinemia, dyslipidemia and metabolic syndrome in adolescents with PCOS was 11.7%, 46.9%, 29.7%, 22.7% and 4.7%, respectively. 16.3%, 74.4%, 67.4%, 39.5% and 14% of the PCOS subjects with BMI > 85th had pre-diabetes, insulin resistance, hyperinsulinemia, dyslipidemia and metabolic syndrome, whereas 9.4%, 32.9%, 10.6%, 14.1% and 0% of the PCOS subjects with BMI < 85th had such disturbances. CONCLUSIONS: Adolescents with PCOS in South China had more metabolic abnormalities than their age- and BMI-matched non-PCOS counterparts. Obesity could worsen insulin resistance, hyperinsulinemia and metabolic syndrome in PCOS adolescents.

Elevated serum anti-mullerian hormone in adolescent and young adult Chinese patients with polycystic ovary syndrome.

PURPOSES: To explore the serum anti-mullerian hormone (AMH) level in adolescent and young adult Chinese patients with polycystic ovary syndrome (PCOS), and to evaluate its diagnostic value for PCOS. STUDY DESIGN: Serum AMH was measured in a cohort of 47 adolescent and young adult Chinese patients with PCOS and 40 age-matched controls. Its diagnostic potential was evaluated by Receiver Operating Characteristic (ROC) curves. RESULTS: The serum AMH level was higher in PCOS patients than in controls (9.85 ± 4.93 ng/mL vs. 7.13 ± 3.02 ng/mL, P = 0.002), and positively related to the mean ovarian volume in PCOS patients (r = 0.319, P = 0.029). The area under the ROC curve for AMH reached a value of 0.664 (0.551-0.778, 95% confidence interval). The best compromise between specificity (70%) and sensitivity (61.7%) was obtained with a cut-off value of 8 ng/mL. CONCLUSIONS: Serum AMH levels are elevated in adolescent and young adult Chinese patients of PCOS. Serum AMH measurement offers a relatively poor diagnostic potency with a sensitivity of 61.7% and a specificity of 70% at 8 ng/mL.

Appropriate BMI levels for PCOS patients in Southern China.

BACKGROUND: This study was designed to establish appropriate body mass index (BMI) level for PCOS women and to compare metabolic abnormalities between PCOS women and control women in Southern China. METHODS: This clinical cross-sectional study included 999 PCOS patients (857 adults and 142 adolescents) and 922 age-matched controls (742 adults and 180 adolescents). RESULTS: The proportion of PCOS patients with a BMI above 23 kg/m(2) was 34.63%. Serum insulin, triglycerides, waist and waist/hip (W/H) measurements were all correlated positively with BMI in PCOS women. Prevalence of metabolic syndrome in Chinese PCOS patients was 18.9%. Receiver-operating characteristic analysis suggested that at the point of BMI > or =23 kg/m(2), the diagnostic power to detect metabolic disorders was at its best. CONCLUSIONS: PCOS patients with a BMI of 23 kg/m(2) or beyond may have a higher risk of metabolic disorders. Using an appropriate BMI level may help to improve attention to metabolic disorders among PCOS patients in Southern China.

[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.

Regulation of gonadotropin-releasing hormone (GnRH) receptor-I expression in the pituitary and ovary by a GnRH agonist and antagonist.

The aim of the current study was to examine the effects of gonadotropin-releasing hormone agonist (GnRH-a) and antagonist (GnRH-ant) on the expression of GnRH receptor-I (GnRHR-I) in pituitary and ovaries in cyclophosphamide (CTX) chemotherapeutic rats and to investigate the possible mechanism of interventions of GnRH-a and GnRH-ant in CTX-induced ovarian damage. In total, 36 female rats were distributed into 6 groups at random: normal saline (NS) group, CTX group, GnRH-a + NS group, GnRH-a + CTX group, GnRH-ant + NS group, and GnRH-ant + CTX group. After the rats were killed, the expression of GnRHR-I messenger RNAs (mRNAs) and proteins in pituitary and ovaries were examined by reverse transcriptase polymerase chain reaction (RT-PCR) and Western blot, respectively. The distribution of GnRHR-I in various compartments of the ovaries was observed by immunohistochemistry. Significant downregulation of GnRHR-I mRNA and protein expression in the pituitary were observed after treatment with GnRH-a or GnRH-ant. Moreover, GnRH-ant was more potent for this direct and fast inhibition. In ovary, GnRHR-I expression was significantly downregulated by GnRH-a. Although GnRH-ant slightly decreased the ovarian expression of GnRHR-I mRNA, the protein level was only weakly changed. In the ovarian compartment, GnRHR-ant groups had markedly GnRHR-I expression in early and late growing follicles compared to GnRHR-a groups that exhibited decreased expression of GnRHR-I, especially in late growing follicles. In summary, this study presents evidence for the different regulating effects of GnRH-a and GnRH-ant on the expression of GnRHR-I in pituitary and ovaries, which may provide insight into the mechanism of GnRH-a and GnRH-ant interventions on chemotoxic ovarian damage.

Association of biochemical hyperandrogenism with type 2 diabetes and obesity in Chinese women with polycystic ovary syndrome.

OBJECTIVE: To evaluate the effect of hyperandrogenism on metabolic disorders among patients with polycystic ovary syndrome (PCOS) diagnosed using the Rotterdam criteria. METHODS: A retrospective analysis of the clinical records of 883 women with PCOS and 717 premenopausal controls identified from the general population. RESULTS: A total of 686 (77.7%) patients were classified with PCOS based on National Institutes of Health (NIH) criteria, and 164 out of 197 (83.2%) additional patients had no hyperandrogenism. Women with normal androgen levels exhibited lower frequencies of obesity, type 2 diabetes, acanthosis nigricans, genetic history of diabetes, and elevated Matsuda index compared with hyperandrogenic patients. Hyperandrogenemia, but not hirsutism, was independently associated with the risk for type 2 diabetes (odds ratio [OR] 5.7; P=0.028) and obesity (OR 1.7; P=0.005) among Chinese patients with PCOS. CONCLUSIONS: Hyperandrogenemia is associated with type 2 diabetes and obesity in Chinese women with PCOS and should be considered at first-line management of hyperandrogenism and infertility due to PCOS.

Increased plasma metastin levels in adolescent women with polycystic ovary syndrome.

OBJECTIVE: This study was designed to: (1) measure metastin levels in women with polycystic ovary syndrome (PCOS) and in adolescent controls; (2) investigate the possible correlations between metastin and PCOS-related reproductive and metabolic disturbances. STUDY DESIGN: The study was a clinical study. Nineteen adolescent women with PCOS, twenty-three adult women with the syndrome, and twenty adolescent controls were selected. Blood samples were collected between day 1 and day 5 of a spontaneous bleeding episode in the PCOS groups and of a menstrual cycle of the controls at 9 a.m. after an overnight fast. Circulating levels of LH, FSH, prolactin, testosterone (T), free testosterone, DHEAS, sex hormone-binding globulin, insulin, glucose and metastin were measured. RESULT(S): Plasma metastin levels are increased in adolescent women with PCOS compared to adolescent controls. Plasma metastin levels were positively correlated with LH levels, 2-h glucose levels and T levels. CONCLUSION(S): These results indicate that metastin is increased in adolescent PCOS women. The increased metastin levels were positively correlated with LH and T levels, and may affect the development of PCOS in adolescents.

[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.

Low prevalence of the metabolic syndrome but high occurrence of various metabolic disorders in Chinese women with polycystic ovary syndrome.

OBJECTIVE: Variations in the prevalence of metabolic syndrome (MetS) among women with polycystic ovary syndrome (PCOS) in different races were reported. We sought to report this prevalence and its components in Chinese women with PCOS and compared these characteristics with healthy controls. DESIGN: Anthropometric measurements and biochemical parameters were evaluated in 578 PCOS patients diagnosed by the Rotterdam criteria and 281 age- and body mass index (BMI)-matched controls. International Diabetes Federation criteria for MetS were used. RESULTS: The prevalence of MetS was 16.8% in this study, and 60.7% of patients displayed at least one component of MetS. Among the patients, the rates of dyslipidemia, impaired fasting glucose, and elevated blood pressure were 41.6, 19.8, and 16.1% respectively; the rates of these corresponding components in age- and BMI-matched controls were 14.6, 5.3, and 5.7% respectively. In PCOS patients, the prevalence of MetS was 0.0, 3.9, 20.2, and 51.1% for four different BMI groups respectively; the prevalence of MetS was 7.3, 14.9, 24.2, and 42.4% in the four age groups respectively. Nearly 90% of patients diagnosed with MetS belonged to overweight and obese groups. BMI and age rather than free testosterone, free androgen index, fasting insulin, or sex hormone-binding globulin were included in formulation for predicting MetS according to multivariable logistic regression. CONCLUSIONS: Low prevalence of MetS but high occurrence of various metabolic disorders was found in women with PCOS compared with age- and BMI-matched controls in this study. BMI and age appeared to contribute more to developing MetS than other parameters associated with insulin resistance or hyperandrogenism.

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.

Cryopreservation of human ovarian tissue by solid-surface vitrification.

OBJECTIVE: To cryopreserve human ovarian tissue using solid-surface vitrification (SSV) technique for the first time. STUDY DESIGN: Human ovarian slices from each of 26 patients were randomly allocated to fresh, SSV, and slow-freezing groups, respectively. Histological observation and the TUNEL assay of the tissue were performed after cryopreservation. In vitro culture was done to study the initial recruitment of follicles and hormone production ability after SSV/slow-freezing. RESULTS: The majority of primordial follicles were maintained intact through either SSV or the slow-freezing method. No statistically significant destructive effect of SSV or slow-freezing for primordial follicles and stromal cells was found using the TUNEL assay. In the SSV and slow-freezing groups, estradiol and progesterone were secreted continuously during 10 days in culture, and the proportions of growing follicles increased significantly comparing to the uncultured fresh group. The follicular proportions and the concentrations of estradiol and progesterone exhibited no statistically significant differences between the SSV and slow-freezing groups. CONCLUSION: SSV is an effective, simple and inexpensive alternative for human ovarian tissue cryopreservation.

Prevalence of polycystic ovary syndrome in unselected women from southern China.

OBJECTIVE: The objective was to determine the prevalence and metabolic parameters of polycystic ovary syndrome (PCOS) in southern China. STUDY DESIGN: The study was observational with a parallel study. Participants were studied in a medical examination center. A population of 915 women of reproductive age was investigated at the time of their annual physical examination to determine the prevalence of PCOS in unselected women from southern China. RESULTS: Our results demonstrated a 2.2% (20/915) prevalence of PCOS. Women with PCOS had higher levels of luteinizing hormone and higher luteinizing hormone/follicle-stimulating hormone ratios than those in the other groups. Women with PCOS had higher fasting insulin levels and lower fasting glucose/fasting insulin ratios than women in any of the other groups. CONCLUSIONS: Some clinical and biochemical characteristics were apparent in PCOS patients in our population, and ethnic differences may be considered when studying the clinical and metabolic features of PCOS in China.

[Effects of gonadotropin releasing hormone analogues on chemotherapy-induced ovarian function damage in rats].

OBJECTIVE: To investigate the effects of gonadotropin releasing hormone agonist (GnRH-a) and antagonist (GnRH-ant) on cyclophosphamide (CTX)-induced ovarian damage in rats. METHODS: Seventy-two Sprague-Dawley female rats were divided randomly into six groups, which received normal saline (NS), CTX, GnRH-a + NS, GnRH-a + CTX, GnRH-ant + NS, and GnRH-ant + CTX respectively. Levels of serum follicle-stimulating hormone (FSH), luteinizing hormone (LH) and estradiol (E(2)) were measured successively by the enzyme-linked immunosorbent assay method, and half of the rats were killed in the first week and between the fourth and the fifth week after stop of medication, respectively to compare the weight of the ovaries and the number of the primordial follicles and the growth follicles. RESULTS: (1) Throughout experiment, the serum levels of FSH, LH and E(2) of the control group fluctuated slightly, while those in the CTX group kept rising. During medication treatment, compared with the control group [(118 +/- 16) microg/L, (350 +/- 35) microg/L] and the CTX group [(113 +/- 15) microg/L, (289 +/- 42) microg/L], the concentrations of LH [(42 +/- 8) - (47 +/- 7) microg/L, (31 +/- 5) - (36 +/- 7) microg/L] and FSH [(124 +/- 45) - (136 +/- 32) microg/L, (178 +/- 54) - (198 +/- 27) microg/L] in the GnRH-a groups and the GnRH-ant groups were maintained at low levels significantly and the levels of LH in the GnRH-ant groups were significantly lower than that in the GnRH-a groups, but the levels of FSH in the GnRH-ant groups were significantly higher than that in the GnRH-a groups (P < 0.05); and extremely different from the GnRH-a groups [(0.98 +/- 0.18) - (1.46 +/- 0.22) ng/L], the level of E(2) of the GnRH-ant groups [(3.58 +/- 0.43) - (3.98 +/- 0.74) ng/L] did not significantly decrease (P < 0.01). After stop of therapy, the concentrations of LH, FSH and E(2) in the GnRH-a groups and the GnRH-ant + NS group rose gradually and were similar to the levels of the control group (P > 0.05), but the levels of FSH, LH and E(2) of the GnRH-ant + CTX group rose obviously and were similar to the levels of the CTX group, especially the FSH , and the levels of LH and FSH of the GnRH-ant + CTX group [(156 +/- 12) microg/L, (520 +/- 44) microg/L] and the CTX group [(178 +/- 18) microg/L, (546 +/- 36) microg/L] were significantly higher than that of the other four groups [(121 +/- 15) - (132 +/- 13) microg/L, (335 +/- 35) - (359 +/- 26) microg/L] at the 4(th) - 5(th) week after stop of treatment (P < 0.05). (2) At the 1(st) week after stopping therapy, the GnRH-a + NS group [(12 +/- 5) mg/100 g]and the GnRH-a + CTX group [(18 +/- 8) mg/100 g] had the lowest weight of ovaries which was significantly different from the other groups [(30 +/- 9) - (37 +/- 8) mg/100 g, P < 0.05]. At the 4(th) - 5(th) week after stopping therapy, the GnRH-ant + CTX group [(22 +/- 6) mg/100 g] and the CTX group [(20 +/- 4) mg/100 g] had the lowest weight of ovaries which were significantly different from the other groups [(29 +/- 5) - (31 +/- 9) mg/100 g, P < 0.05]. (3) At the 1(st) week after stopping therapy, there were the largest number of primordial follicles [(824 +/- 45), (689 +/- 39)] and the lowest number of growth follicles [(15 +/- 1), (21 +/- 3)] in the GnRH-a + NS group and the GnRH-a + CTX group, but there were the lowest number of primordial follicles [(255 +/- 24), (343 +/- 29)] and the largest number of growth follicles [(110 +/- 21), (87 +/- 17)] in the GnRH-ant + CTX group and the CTX group. At the 4(th) - 5(th) week after stopping therapy, the number of growth follicles in the GnRH-a + NS group (58 +/- 11) and the GnRH-a + CTX group (56 +/- 16) recovered to almost the level of the control group (57 +/- 9, P > 0.05), but the number of all kinds of follicles declined significantly in the GnRH-ant + CTX group [(195 +/- 15), (36 +/- 12)] and the CTX group [(212 +/- 11), (36 +/- 9)] compared to the other four groups [(302 +/- 15) - (690 +/- 43), (44 +/- 12) - (58 +/- 11), P < 0.05]. CONCLUSION: In rat model, GnRH-a prevents the ovarian function damage induced by CTX, but GnRH-ant does not show similar protective effect.

[Cyclophosphamide-induced rat ovarian damage and expression of gonadotropin-releasing hormone receptor in the damaged ovaries].

OBJECTIVE: To investigate ovarian follicular damage induced by chemotherapeutic agents and gonadotropin- releasing hormone receptor (GnRHR) expression in the damaged ovaries in rats. METHODS: Two groups of adult SD rats were subjected to intraperitoneal injection of a single-dose cyclophosphamide and saline, respectively, and 8 weeks later, the ovaries were taken for observing the ovarian damages. The distribution of GnRHR was detected with immunohistochemistry, and RT-PCR was used to determine the expression of GnRHR mRNA in the rat ovaries. RESULTS: Massive primordial follicular loss occurred in the ovaries of rats exposed to cyclophosphamide with also evident stromal ovarian blood vessel damages and focal fibrosis. Both the protein and mRNA expressions of GnRHR were detected in normal rat ovaries, but in rats exposed to cyclophosphamide, the expressions were significantly lowered in the ovaries (P<0.05). CONCLUSION: Low-level GnRHR expressions in the ovaries of rats with cyclophosphamide exposure suggest microenvironment disturbances in the damaged rat ovaries in advanced stage of chemotherapy.