Tubal ectopic pregnancy occurrence is associated with high expressions of prokineticin receptors and aberrant secretion of inflammatory cytokines.

OBJECTIVE: Tubal ectopic pregnancy (TEP) remains the most common cause of maternal morbidity and mortality in the early months of pregnancy. The aim of this study is to perform the correlation between PROKRs and pro-inflammatory genes and explore the role of novel genes in pathogenesis of TEP. METHODS: Here, quantitative real time PCR and immunohistochemistry were used to assess the expression of the novel genes in 120 TEP patients and 30 age-matched non-TEP patients. The correlation between PROKRs and pro-inflammatory genes were analyzed by Pearson correlation coefficient. Univariate and multivariate Cox regression analyses were used to assess the risk prediction rate of novel genes. Receiver operating characteristic was used to assess the performance of our model. RESULTS: PROKRs (PROKR1 and PROKR2) and pro-inflammatory genes (TNF-α, IL-6, and IL-8) expression levels significantly enhanced in TEP patients, and significantly positive correlation with pro-inflammatory genes for PROKRs. A multivariate Cox regression analysis demonstrated that 2 genes (PROKR2 and IL8) had significant diagnostic value, which were associated with the occurrence and development of TEP. CONCLUSION: Our data further denote that dysregulation of PROKR2 and IL-8 were risk factor and played an important role in the pathogenesis of TEP.

Novel Epigenomic Biomarkers of Male Infertility Identified by Methylation Patterns of CpG Sites Within Imprinting Control Regions of H19 and SNRPN Genes.

Male infertility is an important global health burden that can benefit from novel biomarkers and diagnostics innovation. Aberrant methylation of the imprinted genes H19 and SNRPN (small nuclear ribonucleoprotein polypeptide N) in sperm DNA has been implicated in abnormal sperm parameters and male infertility. However, whether certain methylation patterns of one or multiple CpG sites within an imprinted gene are pathological for multiple sperm defects remains poorly understood. To examine the diagnostic potential of certain methylation patterns of CpG sites for multiphenotype defects in human sperm, the sperm DNA methylation patterns of individual CpG sites within imprinting control regions (ICRs) of imprinted genes H19 and SNRPN were measured by bisulfite pyrosequencing in a Han Chinese population sample: 39 oligoasthenozoospermia (OA) patients, 36 asthenoteratozoospermia (AT) patients, and 50 normozoospermia (N) controls. A partial least squares discriminant analysis model was built with the CpG sites as independent variables. Among the 16 CpG sites screened, the methylation patterns of eight CpG sites within H19-ICR (CpG sites 1, 6-9, 12 and 15-16), and eight CpG sites within SNRPN-ICR (CpG sites 2, 5-6, 8-10, 13, and 16) correctly classified 74.4% and 72.0% of the samples in terms of male fertile status, respectively. Furthermore, by combination of these 16 selected CpG sites within ICRs of H19 and SNRPN, 88.0% of the samples could be successfully classified. Our study demonstrates that methylation profiles of CpG sites within ICRs of imprinted genes H19 and SNRPN may potentially serve as epigenomic biomarkers for assessment of infertility in men with multiple sperm defects. Further studies in independent population samples are called for diagnostic significance of methylation patterns of CpG sites within imprinted genes.

Functional investigation on aromatase in endometrial hyperplasia in polycystic ovary syndrome cases.

OBJECTIVE: To explore the possible significance of aromatase P450 in endometrial hyperplasia with a background of polycystic ovary syndrome (PCOS). METHODS: Immunohistochemistry was used to determine the expression of aromatase P450 in endometrium of PCOS patients. Semiquantitative analysis of aromatase P450 expression of mRNA and protein level wasalso carried out by real-time quantitative RT-PCR method. After endometrial cells were stimulated by testosterone and letrozole in vitro, the estradiol (E2) level was determined, and the expression of cell aromatase P450 mRNA was assessed. RESULTS: The aromatase P450 mRNA level was increased in endometria of PCOS patients. When endometrial cells were cultured with 10-6 M testosterone, the E2 level in the culture medium increased. An inhibitory effect on E2 generation and expression of aromatase P450 mRNA was observed when the endometrial cells were treated with 10(-5) M letrozole. CONCLUSIONS: There is an increased expression of aromatase P450 in PCOS patient endometrium. Androgen stimulation could enhance the synthesis of aromatase P450 mRNA and the production of E2 in endometrial cells in vitro while letrozole could do the reverse.

Effects of oocyte vitrification on histone modifications.

Vitrification has been widely used as an assisted reproductive technology in animals and humans, yet the impact of oocyte vitrification and warming on survival and histone modifications has to be evaluated. In the present study, the survival of mouse MII oocytes was assessed after freezing, as were changes in histone 3 lysine 9 (H3K9) dimethylation, histone 4 lysine 5 (H4K5) acetylation and histone 3 lysine 14 (H3K14) acetylation. The results show that, in oocytes subjected to vitrification, H3K9 methylation and H4K5 acetylation were increased. H3K14 acetylation could not be detected in either non-vitrified or vitrified oocytes. Oocytes are very sensitive to changes in H3K9 and H4K5 following vitrification. Both these histone modifications could be useful markers to monitor epigenetic perturbations induced by various experimental vitrification protocols and eventually for optimising the cryopreservation of human oocytes.

Selective impairment in glycogen synthase kinase-3 and mitogen-activated protein kinase phosphorylation: comparisons with the hyperandrogenic and the hyperinsulinemic rats.

OBJECTIVE: To characterize and compare the effect of DHEA and insulin plus hCG on ovarian morphology, estrous cycle, hormonal levels, insulin sensitivity, and the regulation of insulin signaling in rats. DESIGN: Animal model study. SETTING: University laboratory. ANIMAL(S): Female Sprague-Dawley rats. INTERVENTION(S): Female rats received DHEA or insulin plus hCG by continuous administration. MAIN OUTCOME MEASURE(S): Ovarian morphology, estrous cycle, hormonal levels, insulin sensitivity, protein levels, and phosphorylation state of glycogen synthase kinase-3beta and extracellular regulated kinase 1/2 in the ovary. RESULT(S): Rats treated with DHEA displayed anovulation, insulin resistance, and polycystic ovaries characterized by cysts and a diminished granulosa layer. In contrast, insulin plus hCG results in acyclicity with increasing androgen biosynthesis and ovarian morphology different from that in DHEA-treated rats. Moreover, we found that insulin-stimulated serine-phosphorylation of glycogen synthase kinase-3beta was higher in insulin plus hCG-treated rats but lower in DHEA-treated rats. Furthermore, basal and insulin-stimulated tyrosine-phosphorylation of extracellular regulated kinase 1/2 was higher in DHEA-treated rats than in controls. CONCLUSION(S): Notwithstanding that both the hyperandrogenism and the hyperinsulinemia synergistic with hCG-treated rats displayed the typical traits of human polycystic ovary syndrome, there is a divergence in the insulin-signaling pathway in the ovarian tissue, which may have a role in the pathogenesis of polycystic ovary syndrome.