Huan Shen

Endometrial MicroRNA Signature during the Window of Implantation Changed in Patients with Repeated Implantation Failure

Background: At present, a diagnostic tool with high specificity for impaired endometrial receptivity, which may lead to implantation failure, remains to be developed. We aimed to assess the different endometrial microRNA (miRNA) signatures for impaired endometrial receptivity by microarray analysis. Methods: A total of 12 repeated implantation failure (RIF) patients and 10 infertile patients, who conceived and delivered after one embryo transfer attempt, were recruited as RIF and control groups, respectively. Endometrial specimens from the window of implantation (WOI) were collected from these two groups. MiRNA microarray was conducted on seven and five samples from the RIF and control groups, respectively. Comparative, functional, and network analyses were performed for the microarray results. Quantitative real-time polymerase chain reaction (PCR) was performed on other samples to validate the expression of specific miRNAs. Results: Compared with those in the control group, the expression levels of 105 miRNAs in the RIF group were found to be significantly up- or down-regulated (at least 2-fold) by microarray analysis. The most relevant miRNA functional sets of these dysregulated miRNAs were miR-30 family, human embryonic stem cell regulation, epithelial-mesenchymal transition, and miRNA tumor suppressors by tool for annotations of microRNA analysis. Network regulatory analysis found 176 miRNA-mRNA interactions, and the top 3 core miRNAs were has-miR-4668-5p, has-miR-429, and has-miR-5088. Expression levels of the 18 selected miRNAs in new samples by real-time PCR were found to be regulated with the same trend, as the result of microarray analysis. Conclusions: There is a significant different expression of certain miRNAs in the WOI endometrium for RIF patients. These miRNAs may contribute to impaired endometrial receptivity.

Diverse endometrial mRNA signatures during the window of implantation in patients with repeated implantation failure

Abstract High endometrial receptivity in the window of implantation (WOI) is essential for successful implantation. However, a diagnostic tool with high specificity for impaired endometrial receptivity remains to be developed. We collected endometrium specimens during the WOI from patients with RIF and women who conceived after one IVF/ICSI attempt. We conducted mRNA microarray on the samples followed by relevant comparative and functional analysis. Microarray analysis revealed 357 dysregulated mRNAs between the two groups. The majority of these mRNAs were found to encode membrane proteins by Gene Ontology (GO) analysis. The major functional biological pathways associated with the down-regulated mRNAs were cytokine-cytokine receptor interaction, the p53 signalling pathway and the complement and coagulation cascades. Up-regulated mRNAs were found mainly to participate in pathways such as PPAR signalling, hematopoietic cell lineage, phosphatidylinositol signalling system, ECM-receptor interaction and notch signalling. AQP3, DPP4 and TIMP3 whose expression patterns were down-regulated in RIF patients both by microarray and real-time PCR had a high correspondence with previous studies demonstrating that these genes may contribute to the defects in endometrial receptivity in RIF patients. Overall, these RIF-associated mRNAs may help devise new diagnostic tools for endometrial receptivity.

Aberrantly expressed long noncoding RNAs in recurrent implantation failure: A microarray related study

ABSTRACT Long noncoding RNAs (lncRNAs) are a class of noncoding RNAs longer than 200 nucleotides. They were long regarded as transcription noise for their low expression and non-protein coding features. Recent published reports indicate that lncRNAs are involved in virtually every aspect of human biology. We aimed to profile the endometrial lncRNA expression pattern in women with recurrent implantation failure (RIF) and predict the function of the genes of the dysregulated lncRNA transcripts. Endometrial samples (24) were collected during window of implantation (14 RIF women and 10 women who conceived after embryo transfer). For the microarray study, 7 RIF endometrium and 5 control endometrium were selected, and quantitative real-time PCR (RT-qPCR) was performed on the rest of the endometrial samples to validate the microarray results. After that, lncRNA-mRNA co-expression analysis, GO analysis, KEGG analysis, and lncRNA-transcript factor (TF) analysis were carried out to analyze the gene functions of the dysregulated lncRNA transcripts. We detected a total of 197 lncRNA transcripts that were dysregulated in RIF endometrium compared with the control group. The relative expression levels of eight selected lncRNA transcripts were validated by RT-qPCR and were in accordance with the microarray outcomes. GO and KEGG analyses revealed that the coexpressed mRNA transcripts were involved in pathways that may affect endometrial receptivity such as cell adhesion. The lncRNA target predictions provided potential TF targets of the dysregulated lncRNA transcripts. Our results indicate that lncRNA expression profiles of RIF endometrium were different from that of normal receptive endometrial, suggesting that lncRNAs may regulate endometrial receptivity. Abbreviations: GO: Gene Oncology; GFs: growth factors; KEGG: Kyoto Encyclopedia of Genes and Genomes; lncRNAs: long noncoding RNAs; PCA3: prostate cancer antigen 3; RT-qPCR: quantitative real-time PCR; RIF: recurrent implantation failure; STK: serine/threonine kinase; TF: transcription factor; WOI: window of implantation

Correlation between receptor-interacting protein 140 expression and directed differentiation of human embryonic stem cells into neural stem cells

Overexpression of receptor-interacting protein 140 (RIP140) promotes neuronal differentiation of N2a cells via extracellular regulated kinase 1/2 (ERK1/2) signaling. However, involvement of RIP140 in human neural differentiation remains unclear. We found both RIP140 and ERK1/2 expression increased during neural differentiation of H1 human embryonic stem cells. Moreover, RIP140 negatively correlated with stem cell markers Oct4 and Sox2 during early stages of neural differentiation, and positively correlated with the neural stem cell marker Nestin during later stages. Thus, ERK1/2 signaling may provide the molecular mechanism by which RIP140 takes part in neural differentiation to eventually affect the number of neurons produced.

Impact of surgery for endometriomas on pregnancy outcomes following in vitro fertilization-intracytoplasmic sperm injection. Who should be the preferred laparoscopists: gynecologists or reproductive surgeons?

ObjectiveTo investigate whether laparoscopic excision of ovarian endometriomas pretreated with operation by gynecologists or reproductive surgeons exerts different effects on in vitro fertilization-intracytoplasmic sperm injection results.Materials and methodsRetrospective case control study. Relevant information was collected from the electronic records of women who underwent IVF/ICSI from 01/01/2013 to 30/12/2015 in our unit. The study group consisted of 35 women who previously had laparoscopic endometrioma excision by reproductive surgeons in our unit; the control group included 36 patients who underwent surgery for endometriomas by gynecologists in our hospital.Result(s)There were slightly higher numbers of AFC and higher pregnancy rate in the study group, although differences did not reach statistical significance. For patients over 35 years old, there were more oocyte retrieved, mature oocytes and two pronucei (2PN) in the study group than the control group although observed differences did not reach statistical significance.Conclusion(s)Electrocautery is more deleterious on ovarian reserve than hemostatic suture. In procedure of patients who wish to conceive, surgeons should use hemostatic suturing technique preferentially.

The Effect of Mono-(2-Ethylhexyl) Phthalate on Human Granulosa Cells

Various phthalates are used extensively as plasticizers in kinds of consumer products. They are harmful for human reproduction and fertility. Mono-(2-ethylhexyl) phthalate (MEHP) is the active metabolite of di-(2-ethylhexyl) phthalate(DEHP),which is most commonly used as a plasticizer and solvent in numerous consumer products. MEHP has been demonstrated to be a reproductive toxicant in rodents, but the evidences of using human cells are rare. In present study, we examined the effect of MEHP on viability and steroid production of human granulosa cells. Human granulosa cells obtained from nine patients undergoing in vitro fertilization were cultured in medium with various concentrations of MEHP (100,300,500 µmol L− 1). After culture for 48 hours, granulosa cells viability was determined. Estradiol, progesterone and testosterone were assayed in the spent culture medium. We found that the viability of granulosa cells was reduced obviously at 100, 300 and 500µmol L− 1 of MEHP. High concentration of MEHP(500uM) was harmfull to steriod hormone synthesis, such as estradiol and progestorone. These findings will provide favorable evidence for adverse effect of MEHP on womens reproductive system.

The Pilot Study of Phthalates and Monoesters in Human Chorion Tissues

Phthalates are widely used in industry as solvents, additives and plasticizers. Some phthalates and their monoester metabolites are known to cause carcinogenic, toxic or endocrine-modulating effects. Chorion tissues as human early embryo are the base of organogenesis. Embryo (3-8 weeks) is sensitive period of teratogensis. With the development of industrialization, in addition to genetic factors, environmental factors become an important reason for human reproductive system defect, particularly under the interaction of the environmental factors and genetic factors. Recently we have studied human chorion tissues to see whether the phthalates especially their monoester metabolites are exist in. In the result five phthalates and monoester metabolites such as dibutyl phthalate (DBP), butyl benzyl phthalate (BBP), di(2-ethylhexyl) phthalate (DEHP), monobutyl phthalate(MBP) and mono(2-ethyl-hexyl) phthalate(MEHP) are discovered, that means that phthalates and their monoesters both can pass through embryo and affect fetus before birth.