Follicular hyperandrogenism and insulin resistance in polycystic ovary syndrome patients with normal circulating testosterone levels

Polycystic ovary syndrome（PCOS） is a common reproductive disease with high heterogeneity. The role of excess androgen in PCOS etiology remains disputed, since around 20%-50% of PCOS women do not display hyperandrogenemia. The microenvironment of the ovary critically influences follicular development. In the present study, we assessed the role of androgen in PCOS by investigating whether excessive follicular fluid androgen was present in PCOS patients with normal serum androgen levels and influenced by follicular fluid insulin resistance（IR）.Follicular fluid samples of 105 women with PCOS and 105 controls were collected. Levels of steroid hormones,glucose and insulin in the follicular fluid were examined and compared with data from serum biochemistry tests. We found that 64.9%（63/97） of PCOS patients with normal serum androgen levels displayed abnormally high follicular fluid androgen level. The follicular fluid androgen level was positively correlated with follicular fluid IR within a certain range and follicular fluid estrogen-to-testosterone（E2/T） ratio was significantly reduced in these patients.These results indicated that there existed a subgroup of PCOS patients who displayed excessive follicular fluid androgen and IR despite their normal circulating testosterone（T） levels. Our study highlights the importance of ovary hyperandrogenism and IR in the etiology of PCOS.

Proteome changes of porcine follicular fluid during follicle development

Background:Ovarian follicular fluid influences follicle and oocyte growth,but the fluctuation of its protein content during folliculogenesis has not been comprehensively analyzed.Here we used a shotgun approach and bioinformatics analyses to investigate and compare the proteomes of porcine follicular fluid(pFF)obtained from small(<4 mm),medium(4–6 mm)and large(>6–12 mm)follicles.Results:Follicular fluid samples containing highest estrogen levels were selected as non-atretic from small(SNA:26.1±15 ng/mL),medium(MNA:162±54 ng/mL),and large(LNA:290±37 ng/mL)follicles for proteomic analyses.We detected 1627,1699,and 1756 proteins in SNA,MNA,and LNA samples,respectively.Nearly 60–63%of total proteins were specific to each sample,11–13%were shared in pairwise comparisons,and 247 proteins were shared among all samples.Functional categorization indicated comparable gene ontology(GO)terms distribution per cellular component,molecular function,and biological process categories across samples;however,the ranking of highly significantly enriched GO terms per category revealed differences between samples.The patterns of proteinto-protein interactions varied throughout follicle development,and proteins such as serine protease inhibitor,clade E(SERPINE);plasminogen activator,urokinase(PLAU);and plasminogen activator,urokinase receptor(PLAUR)appeared stage-specific to SNA,MNA,and LNA,respectively.The“complement and coagulation cascades”was the common major pathway.Besides,properdin and fibulin-1 were abundant proteins that appeared absent in LNA samples.Conclusion:This study provides extensive and functional analyses of the pFF proteome changes during folliculogenesis and offers the potential for novel biomarker discovery in pFF for oocyte quality assessment.

Association of the microbial culture of follicular fluid,vaginal swab and catheter tip with β-hCG IVF positive and negative

Objective:To find out the association of microbial contamination withβ-human chorionic gonadotropin(β-hCG)in-vitro fertilization(IVF)positive and negative.Methods:A total of 73 fresh IVF cycle women were included in the retrospective study.Vaginal swab culture samples were collected prior to ovum pick-up and embryo transfer.The follicular fluids were collected during ovum pick-up and catheter tip culture samples were collected after successful embryo transfer.After 14 days of the embryo transfer,women were classified intoβ-hCG IVF positive and negative.The comparative statistical analyses of aerobic microbial culture reports were done betweenβ-hCG IVF positive and negative women.Results:Out of 73 women,42(57.5%)were found to beβ-hCG IVF positive and 31(43.5%)were negative.In the aerobic culture of ovum pick-up vaginal swab,follicular fluid,embryo transplantation vaginal swab and catheter tip,Enterococcus faecalis was found to be higher compared to other bacteria(Streptococcus spp.,Candida,Escherichia coli and Klebsiella).Regarding the comparison between IVF positive and negative,the overall microbial infection rate of vaginal swab culture during ovum pick-up and embryo transplantation was found to be higher in IVF negative women than in IVF positive women(38.71%vs.28.57%);however,it was not statistically significant(P>0.05).The follicular fluid and catheter tip culture microbial infection rate was found to slightly higher in IVF positive women than in IVF negative women(54.76%vs.41.94%and 19.05%vs.9.68%,respectively),but there were not significant differences(P>0.05).Conclusions:The aerobic microbial culture reports of follicular fluid,vaginal swab culture,and catheter tip culture are not statistically significantly withβ-hCG IVF positive.

Protein Expression of Goat Follicular Fluid （GFF）

Oocyte maturation process is very complex. Until now synthesis and protein function as well as pathway mechanism in oocyte maturation process are not yet well understood. The mechanism of goat follicular fluid （GFF） in maturation in vitro medium needs to be deeply studied before it is implemented widely. The aim of the research was analyzing the GFF to know protein expression of ERK1, ERK 2, p90rsk, Cyclin-Bl and cdc25A by Western Blotting method. GFF collection by aspiration from small follicle （〈 4 mm） and big follicle （4-8 mm） by using needle with spuit size of 18 G. It was concluded that there was protein expression of ERK2 and p90rsk, at 42 kDa and 22 kDa respectively.

Influence of Follicular Fluid on <i>in Vitro</i>Maturation and Fertilization of Bovine Oocytes

The aim of this study was to investigate the effect of time on in vitro maturation of bovine oocytes and of the addition of follicular fluid on meiotic progression. The cumulus-oocyte complexes (COCs) collected from 3 to 6 mm follicles were obtained from ovaries of slaughtered female animals. The medium of maturation was supplemented or not with 20 μL follicular fluid (FF);661 oocytes were matured in vitro (extrusion of the first polar corpuscle) for 22 hours with added follicular fluid (AFF) (72.01%) or without follicular fluid (WFF) (67.53%) and 679 oocytes were matured in vitro for 26 hours (extrusion of the first polar corpuscle) with AFF (92.1%) and WFF (77.15%). The results of extrusion of the second polar corpuscle as an event related to the fertilization percentages showed that the increase in the fertilization rate is maintained at 26 hours with AFF (79.45%), but the percentage decreases WFF (65.08%). After 22 hours, the fertilization rate was 62.38% AFF and 53.40% WFF. The developmental competence of bovine oocytes is affected by the duration of maturation in vitro and the inclusion in the FF culture medium. The use of follicular fluid in the in vitro maturation medium may be a biological strategy to increase the cumulus expansion, the nuclear maturation and the in vitro fertilization.

Follicular fluid levels of prostaglandin E2 and the effect of prostaglandin E2 on steroidogenesis in granulosa-lutein cells in women with moderate and severe endometriosis undergoing in vitro fertilization and embryo transfer

Background The mechanisms of endometriosis with infertility have not been fully studied. The present study aimed to assess the follicular fluid （FF） levels of prostaglandin E2 （PGE2）, which plays a critical role within the ovary, and to investigate the effect of PGE2 on steroidogenesis in granulosa-lutein cells （GLCs） from women with and without endometriosis. Methods Thirty-three women with laparoscopically documented endometriosis and 40 controls undergoing in vitro fertilization （IVF） were studied. We assayed the concentrations of PGE2 in FF, the production of E2 and progesterone in FF and in culture medium, and the expression of steroidogenic acute regulatory protein （STAR） and CYP19A1 in GLCs with the intervention of PGE2. Results PGE2 and progesterone concentrations were increased and displayed positive correlation in endometriotic FE PGE2 induced the expression of STAR and the production of progesterone in GLCs from women with endometriosis, and the expression of STAR and the production of progesterone were increased in GLCs from women with endometriosis. However, there were no significant effects of PGE2 on promoting the production of E2 or the expression of CYP19A1 in GLCs. Moreover, the production of E2 and the expression of CYP19A1 in GLCs from women with endometriosis were significantly decreased compared to the controls. Conclusions PGE2 concentrations are increased in endometriotic FF, along with concomitant increases in progesterone and STAR. In contrast, the E2 and CYP19A1 are decreased in GLCs, which may delay the development of the follicles and cause an imbalance in the follicular steroid hormone levels. These changes may have close relationship with endometriosis-associated infertility.

Effect of Quyu Jiedu Granule(祛瘀解毒颗粒) on Micro-environment of Ova in Patients with Endometriosis

Objective:To observe the effect of Quyu Jiedu Granules(祛瘀解毒颗料,QJG) on the micro-environment of ova in patients with endometriosis(EM).Methods:Twenty EM patients who received in vitro fertilization and embryo transfer(IVF-ET) were randomized equally into a treated group and a control group. Further,20 patients who received IVF-ET due to oviduct factors were enrolled into a non-endometriosis group. The dosage of gonadotrophic hormone used,the number of ova attained,fertilization rate and clinical pregnancy ra...

Effect of soothing liver therapy on oocyte quality and growth differentiation factor-9 in patients undergoing in vitro fertilization and embryo transfer

OBJECTIVE:To investigate the effect of Soothing liver therapy on infertile women undergoing in vitro fertilization and embryo transfer(IVF-ET)and to explore its mechanism.METHODS:Fifty-eight women with tubal infertility were randomized into two groups:30 in an experimental group treated with Xiaoyao powder(Shugan)plus gonadotropin-releasing hormone analog(GnRHa)/follicle-stimulating hormone(FSH)/human chorionic gonadotropin(hCG)and 28 in the control group who were treated with GnRHa/FSH/hCG only.The total gonadotropin(Gn)doses required,endometrial thickness,oocyte numbers,high quality embryo production rate and pregnancy rate of the two groups were compared.The concentration of growth differentiation factor-9(GDF-9)in follicular fluid was detected by western blotting and the expression of GDF-9 mRNA in granulosa cells was measured using reverse tran-scription-polymerase chain reaction amplification.RESULTS:In the experimental group,the Gn dose was significantly lower than that in the control group;the endometrial thickness,high quality embryo production and pregnancy rates were significantly higher and the expression of GDF-9 mRNA was also significantly higher than in the control group(all P<0.05).CONCLUSION:Shugan treatment can improve the pregnancy rate of women with tubal infertility;its mechanism is possibly related to the increased expression of GDF-9 in granulosa cells.