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Early non-chromosome-related missed abortion (MA) is commonly associated with an altered immunological environment during pregnancy. Human decidual natural killer (dNK) cells, the most abundant lymphocyte population within the first-trimester maternal-fetal interface, are vital maternal regulators of immune tolerance mediating successful embryo implantation and placentation. Previous studies have shown that dNK cells may play a role in MA. However, the gene expression status and specific altered manifestations of dNK cells in patients with early MA remain largely unknown. Here, we show that MA dNK cells have distinct mRNA and lncRNA expression profiles through RNA sequencing, with a total of 276 mRNAs and 67 lncRNAs being differentially expressed compared with controls. Protein-protein interaction analysis of differentially expressed mRNAs was performed to identify hub genes and key modules. An lncRNA-mRNA regulatory network characterized by the small-world property was constructed to reveal the regulation of mRNA transcription by differential hub lncRNAs. Functional annotation of differentially expressed mRNAs and lncRNAs was performed to disclose their potential roles in MA pathogenesis. Our data highlight several enriched biological processes (immune response, inflammatory response, cell adhesion, and extracellular matrix [ECM] organization) and signaling pathways (cytokine-cytokine receptor interaction, ECM-receptor interaction, Toll-like receptor signaling pathway, and phosphatidylinositol signaling system) that may influence MA. This study is the first to demonstrate the involvement of altered mRNA and lncRNA expression profiles in the dNK cell pathogenesis of early MA, facilitating a better understanding of the underlying molecular mechanisms and the development of novel MA therapeutic strategies targeting key mRNAs and lncRNAs.

Background Nicolaides–Baraitser syndrome (NCBRS) is a severe neurodevelopmental disorder with multiple abnormalities. To date, all pathogenic variants in SMARCA2 causing NCBRS are de novo and most are missense variants located in the ATPase domain of SMARCA2 protein. Methods In this study, a familial trio whole‐exome sequencing was performed on the proband presenting with intellectual disability, early‐onset epilepsy, and autistic features. A novel missense variant c.553C>G (p.Gln185Glu) in SMARCA2 was identified, which is located in the QLQ domain. The same variant was subsequently also found in the mother's ongoing pregnancy. Samples from accessible tissues such as saliva and sperm other than blood were collected from the parents, and the detection of the target variant was performed by amplicon‐based deep sequencing. Results Low‐level mosaicism of the target variant c.553C>G (p.Gln185Glu) was detected in the father's sperm with allele fraction of 2.8% by amplicon‐based deep sequencing, which was not detected in either parents’ blood or saliva specimens. Heterozygosity of this variant was confirmed in the proband. Conclusion This is the first report of paternal germline mosaicism for a SMARCA2 disease‐causing variant. In addition, the missense variant c.553C>G (p.Gln185Glu) in the QLQ domain causes mainly neurological and developmental phenotypes with unremarkable characteristic facial features and limb abnormalities. Our findings expand the phenotypic spectrum and mode of genetic transmission associated with the SMARCA2 variants., A novel SMARCA2 missense variant in the proband was found due to low‐level paternal germline mosaicism as the first disease‐causing variant in the QLQ domain associated with solely neurological and developmental phenotypes of NCBRS, which provides new insights into the structure–function relationship of this subunit in the SWI/SNF complexes.

Brown adipose tissue (BAT) is an exclusive tissue of nonshivering thermogenesis. It is fueled by lipids and glucose and involved in energy and metabolic homeostasis. Intrauterine exposure to hyperglycemia during gestational diabetes mellitus may result in abnormal fetal development and metabolic phenotypes in adulthood. However, whether intrauterine hyperglycemia influences the development of BAT is unknown. In this study, mouse embryos were exposed to the intrauterine hyperglycemia environment by injecting streptozocin into pregnant mice at 1 d post coitum (dpc). The structure of BAT was examined by hematoxylin and eosin staining and immunohistochemical analysis. The glucose uptake in BAT was measured in vivo by [18F]-fluoro-2-deoxyglucose-micro-positron emission tomography. The gene expression in BAT was determined by real-time PCR, and the 5'-C-phosphate-G-3' site-specific methylation was quantitatively analyzed. Intrauterine hyperglycemia exposure resulted in the impaired structure of BAT and decreased glucose uptake function in BAT in adulthood. The expressions of the genes involved in thermogenesis and mitochondrial respiratory chain in BAT, such as Ucp1, Cox5b, and Elovl3, were down-regulated by intrauterine hyperglycemia exposure at 18.5 dpc and at 16 wk of age. Furthermore, higher methylation levels of Ucp1, Cox5b, and Elovl3 were found in offspring of mothers with streptozotocin-induced diabetes. Our results provide the evidence for enduring inhibitory effects of intrauterine hyperglycemia on BAT development in offspring. Intrauterine hyperglycemia is associated with increased DNA methylation of the BAT specific genes in offspring, which support an epigenetic involvement.-Yu, D.-Q., Lv, P.-P., Yan, Y.-S., Xu, G.-X., Sadhukhan, A., Dong, S., Shen, Y., Ren, J., Zhang, X.-Y., Feng, C., Huang, Y.-T., Tian, S., Zhou, Y., Cai, Y.-T., Ming, Z.-H., Ding, G.-L., Zhu, H., Sheng, J.-Z., Jin, M., Huang, H.-F. Intrauterine exposure to hyperglycemia retards the development of brown adipose tissue.

Objective This study aimed to determine the effects of diet-induced paternal obesity on cognitive function in mice offspring. Methods Male mice (F0) were randomized to receive either a control diet (10 kcal% fat) or a high-fat diet (HFD; 60 kcal% fat) for 10 weeks before being mated with normal females to generate F1 offspring. Male F1 offspring were mated with normal females to generate F2 offspring. Behavioral tests were used to assess cognitive functions in F1 and F2 offspring. Reduced representation bisulfite sequencing was used to the explore mechanisms of epigenetic inheritance. Results HFD-induced paternal obesity resulted in cognitive impairments in F1 offspring, potentially due, at least in part, to increased methylation of the BDNF gene promoter, which was inherited from F0 spermatozoa. BDNF/tyrosine receptor kinase B signaling was associated with cognitive impairments in HFD-fed F1 offspring. However, there were no significant changes in F2 offspring. Conclusions The findings provide evidence of intergenerational effects of paternal obesity on cognitive function in offspring occurring via epigenetic spermatozoan modifications.

Background Diet and exercise during pregnancy have been used to prevent gestational diabetes mellitus (GDM) with some success. Objective To examine the effectiveness of lifestyle intervention on GDM prevention and to identify key effectiveness moderators to improve the prevention strategy. Search strategy Pubmed, Scopus, Cochrane, and cross-references were searched. Selection criteria Randomised controlled trials (RCTs) evaluating lifestyle interventions during pregnancy for GDM prevention. Data collection and analysis Two independent reviewers extracted data. A random-effects model was used to analyse the relative risk (RR) and 95% confidence interval (95% CI). Meta-regressions and subgroup analyses were used to investigate important moderators of effectiveness. Main results Forty-seven RCTs involving 15 745 participants showed that diet and exercise during pregnancy were preventive of GDM (RR 0.77, 95% CI 0.69-0.87). Four key aspects were identified to improve the preventive effect: targeting the high-risk population; an early initiation of the intervention; the correct intensity and frequency of exercise; and gestational weight gain management. Although 24 RCTs targeted women who were overweight or obese, body mass index (BMI) failed to predict the effectiveness of an intervention. Instead, interventions are most effective in high-incidence populations rather than simply in women who are overweight or obese. Furthermore, exercise of moderate intensity for 50-60 minutes twice a week could lead to an approximately 24% reduction in GDM. Conclusion The best strategy to prevent GDM is to target the high-risk population predicted by risk evaluation models and to control the gestational weight gain of women through intensified diet and exercise modifications early in their pregnancy. Tweetable abstract Four key effectiveness moderators of lifestyle interventions for GDM prevention.

Objective Chromosomal abnormalities such as aneuploidy have been shown to be responsible for causing spontaneous abortion. Genetic evaluation of abortions is currently underperformed. Screening for aneuploidy in the products of conception can help determine the etiology. We designed a high-throughput ligation-dependent probe amplification (HLPA) assay to examine aneuploidy of 24 chromosomes in miscarriage tissues and aimed to validate the performance of this technique. Methods We carried out aneuploidy screening in 98 fetal tissue samples collected from female subjects with singleton pregnancies who experienced spontaneous abortion. The mean maternal age was 31.6 years (range: 24–43), and the mean gestational age was 10.2 weeks (range: 4.6–14.1). HLPA was performed in parallel with array comparative genomic hybridization, which is the gold standard for aneuploidy detection in clinical practices. The results from the two platforms were compared. Results Forty-nine out of ninety-eight samples were found to be aneuploid. HLPA showed concordance with array comparative genomic hybridization in diagnosing aneuploidy. Conclusion High-throughput ligation-dependent probe amplification is a rapid and accurate method for aneuploidy detection. It can be used as a cost-effective screening procedure in clinical spontaneous abortions. © 2016 John Wiley & Sons, Ltd.

Diabetic pregnancy, with ever increasing prevalence, adversely affects embryogenesis and increases vasculometabolic disorder risks in adult offspring. However, it remains poorly understood whether maternal diabetes increases the offspring's susceptibility to heart injuries in adulthood. In this study, we observed that cardiac function and structure were comparable between adult offspring born to diabetic mice and their counterparts born to nondiabetic mice at baseline. However, in response to myocardial ischemia/reperfusion (MIR), diabetic mother offspring exhibited augmented infarct size, cardiac dysfunction, and myocardial apoptosis compared with control, in association with exaggerated activation of mitochondria- and endoplasmic reticulum (ER) stress-mediated apoptosis pathways and oxidative stress. Molecular analysis showed that the impaired myocardial ischemic tolerance in diabetic mother offspring was mainly attributable to blunted cardiac insulin receptor substrate (IRS)-1/Akt signaling. Furthermore, the effect of maternal melatonin administration on offspring's response to MIR was determined, and the results indicated that melatonin treatment in diabetic dams during pregnancy significantly improved the tolerance to MIR injury in their offspring, via restoring cardiac IRS-1/Akt signaling. Taken together, these data suggest that maternal diabetes predisposes offspring to augmented MIR injury in adulthood, and maternal melatonin supplementation during diabetic pregnancy may hold promise for improving myocardial ischemic tolerance in the offspring.

It is widely believed that endometrial atrophy in postmenopausal women is due to an age-related reduction in estrogen level. But the role of high circulating follicle-stimulating hormone (FSH) in postmenopausal syndrome is not clear. Here, we explored the role of high circulating FSH in physiological endometrial atrophy. We found that FSH exacerbated post-OVX endometrial atrophy in mice, and this effect was ameliorated by lowering FSH with Gonadotrophin-releasing hormone agonist (GnRHa). In vitro, FSH inhibited endometrial proliferation and promoted the apoptosis of primary cultured endometrial cells in a dose-dependent manner. In addition, upregulation of caspase3, caspase8, caspase9, autophagy-related proteins (ATG3, ATG5, ATG7, ATG12 and LC3) and downregulation of c-Jun were also observed in endometrial adenocytes. Furthermore, smad2 and smad3 showed a time-dependent activation in endometrial cells which can be partly inhibited by blocking the transforming growth factor beta receptor II (TβRII). In conclusion, FSH regulated endometrial atrophy by affecting the proliferation, autophagy and apoptosis of endometrial cells partly through activation of the transforming growth factor beta (TGFβ) pathway.

Aims The aim of this study was to determine the promoter methylation status of type 2 isoform of 11β-hydroxysteroid dehydrogenase (11β-HSD2) and its regulatory correlation with 11β-HSD2 gene expression in placentas of pre-eclampsia (PE) patients of Chinese Han ethnicity. Material and Methods The pathological features of placental tissues were studied using hematoxylin–eosin staining and immunohistochemical staining. The 11β-HSD2 mRNA and protein expressions were detected by real-time polymerase chain reaction and Western blotting. The methylation of the 11β-HSD2 promoter sequence was examined by bisulfite sequencing polymerase chain reaction. Results Trophoblast hyperplasia and discontinuous syncytial layer were observed in the PE group, and the 11β-HSD2 was distributed irregularly and its immunoreactivity was weakened distinctly. The expressions of 11β-HSD2 mRNA and protein decreased significantly in the PE group compared with the control group. Unexpectedly, almost no 11β-HSD2 methylation was detected in PE placental tissue (two fragments, 0.6% vs 0%) or normal placental tissue (1% vs 0.6%). No significant difference in 11β-HSD2 promoter methylation was found between the two groups. Conclusions The 11β-HSD2 expression decreased in PE women of Chinese Han ethnicity, but was not interrelated with the promoter methylation status.

Objective The aim of this study is to report the clinical application of noninvasive prenatal testing (NIPT) to detect chromosomal aneuploidies, especially trisomies 21, 18, and 13 in Chinese singleton pregnancies. Methods Pilot validation between NIPT with full karyotyping was conducted blindly on 306 cases. Subsequently, 7705 pregnancies were offered with NIPT. Follow-up data was obtained in all cases. Results In the validation stage, a total of five NIPT positive cases were observed for trisomies 21 and 18, and results were confirmed by karyotyping; there were no cases of trisomy 13. Thus, the overall sensitivity and specificity in the validation stage was 100%. In 7705 cases, NIPT results were obtained in 7701 cases; 66 cases were classified as positive, including 48 cases of trisomy 21, 14 cases of trisomy 18, and 4 cases of trisomy 13. Subsequent karyotyping documented two false positive diagnoses for trisomies 21, 18, and 13, respectively. Sensitivity and specificity for detection of trisomies 21 and 18 and 13 were 100% and 99.9%, respectively. Additionally, prenatal chromosomal detection for pregnancies with NIPT has shown a gradual increase since its implementation. Conclusion Noninvasive prenatal testing allows a more suitable and efficient workflow for our patients' needs, together withinvasive proceduresallowsa higherprenataldetectionofchromosomal aneuploidies.©2014JohnWiley& Sons,Ltd.

SummaryObjective To study the efficiency of letrozole, an aromatase inhibitor, in endometrial preparation for cryopreserved–thawed embryo transfer (FET) in women with polycystic ovarian syndrome (PCOS). Design Retrospective observational study. Patients One hundred and twenty patients with PCOS who met the inclusion criteria for the study. Measurements We assessed in vitro fertilization outcomes in one hundred and twenty patients with PCOS (148 cycles) who were prepared for and underwent FET between June 2011 and December 2012. Patients were prepared for FET using artificial hormone cycles induced with oestrogen and progesterone supplementation (n = 76), letrozole stimulation (n = 40) or hMG stimulation (n = 32). Results There were no differences in demographic characteristics between the groups, except that the letrozole group had a higher incidence of embryo transfer failure in the past. The letrozole stimulation group had a significantly higher maximal endometrial thickness and significantly higher rates of clinical pregnancy per transfer, ongoing pregnancy per transfer and implantation, compared with the artificial and hMG stimulation groups. Differences in these parameters between the artificial and hMG stimulation groups were not significant. Conclusion Using letrozole stimulation in endometrial preparation for cryopreserved–thawed embryo transfer in patients with PCOS may be associated with better outcomes than using hormonal manipulation or hMG stimulation.

The aims were to clarify the effect of placental trophoblasts on T lymphocytes by assessing production of cytokines and expression of transcription factors regulating Th1, Th2, and Th17 immunity in T lymphocytes. Placental trophoblasts were isolated and conditioned medium was made after trophoblast cultivation for 72 h. T lymphocytes were cultured in presence or absence of conditioned medium. ELISA was used to detect concentration of IL-2, TNF-α, IFN-γ, IL-4, IL-10, and IL-17 in supernatants of T cell and real-time PCR was used to detect the status of Th1 (T-bet, STAT-4), Th2 (GATA-3, STAT-6), and Th17 (RORC) immunity in T lymphocyte. We found that the level of IL-2, IFN-γ, TNF-α, and IL-17 was significantly decreased when T lymphocytes were cultured in conditioned medium compared with control, while IL-10 and IL-4 level were not significantly changed. The presence of conditioned medium significantly decreased the ratio of Th1/Th2. The expression of GATA-3 and STAT-6 were significantly increased and STAT-4 was reduced when T lymphocyte was cultured in conditioned medium, while the expression of T-bet and RORC was not significantly different. We concluded that placental trophoblast-induced shift of Th1/Th2 balance toward Th2 and inhibition of Th17 might be among the mechanisms involved in maternal tolerance to fetus.

Problem l-selectin ligand has displayed mediating adhesion at the maternal–fetal interface. Therefore, we investigated the impact of l-selectin ligand on establishing pregnancy in women undergoing in vitro fertilization and embryo transfer (IVF-ET). Method of study Endometrium between cycle days LH +6 to +9 was obtained from 56 Chinese women referred for IVF and tested for l-selectin ligand by immunohistochemistry and Western blot. The standard gonadotropin-releasing hormone agonist long protocol was used for ovarian stimulation. Results l-selectin ligand was localized in the endometrial gland and luminal epithelial cells. Western blot analysis of endometrium identified four bands and levels of component 1, 2 and 4 were significantly higher in the pregnancy group than in the non-pregnancy group (P

To characterise the changes in serum visfatin levels in late normal pregnancy and pre-eclampsia.Twenty-seven women with pre-eclampsia were recruited. Twenty-eight women in the third trimester of normal pregnancy served as pregnant control and 28 healthy non-pregnant women as non-pregnant control. Serum levels of visfatin were measured with an enzyme-linked immunosorbent assay.The means of serum visfatin were 626.4+/-45.5 ng/ml (mean+/-SEM) in non-pregnant control, 695.9+/-92.5 ng/ml in pregnant control, and 308.3+/-80.0 ng/ml in pre-eclampsia, respectively, and were significantly different among the groups (p0.001). Visfatin level was significantly lower in pre-eclampsia compared to non-pregnant control (p=0.004) and pregnant control (p0.001). Women with severe pre-eclampsia had a significantly lower serum visfatin level than those with mild pre-eclampsia (114.6+/-80.9 versus 425.2+/-122.1 ng/ml, p=0.037).A decrease in visfatin level was demonstrated in pre-eclampsia, suggesting that visfatin and adipokine-associated metabolic abnormalities are involved in the pathogenesis of the disease.

To explore the biological characteristics of the recombinant zona pellucida 3 (ZP3) peptides of rhuZP3a 22,176 and rhuZP3b 177,348 , we examined whether rhuZP3a 22,176 or rhuZP3b 177,348 trigger the acrosome reaction (AR) of human spermatozoa and we investigated the underlying mechanism. The assessment of AR was performed using chlortetracycline staining. The intracellular free calcium concentration ((Ca 2+ )i) in Fura-2/AM- loaded human sperm was monitored with a spectrofluorophotometer. We found that rhuZP3a 22,176 and rhuZP3b 177,348 were capable of eliciting AR at different concentrations. With the addition of either peptide, the (Ca 2+ )i level was raised to a peak with or without a plateau. The AR could be inhibited by pertussis toxin (PTX), EGTA, and pimozide (a T-type calcium channel blocker), whereas verapamil was less effective in this regard. The results of the present study suggest that peptides rhuZP3a 22,176 and rhuZP3b 177,348 have a role similar to human ZP3, and that the mechanism of the response to the peptides involves influx of calcium, the G protein pathway, and a T- type calcium channel.

In the report an adolescent female patient who suffered from continuous lower abdominal distending pain with an ovarian cyst was successfully cured with Chinese herbs. Chinese herbs were orally administered for 18 days, after which all of the symptoms and the ovarian cyst disappeared.