Zhou X.-P.,Key Laboratory of Womens Reproductive Health of Zhejiang Province |
Hu X.-L.,Key Laboratory of Womens Reproductive Health of Zhejiang Province |
Zhu Y.-M.,Brigham and Womens Hospital |
Qu F.,Zhejiang University |
And 2 more authors.
Asian Journal of Andrology | Year: 2011
In this study, we aimed to determine the effects of hepatitis B virus (HBV) infection on sperm quality and the outcome of assisted reproductive technology (ART). A total of 916 men (457 HBV-positive and 459 HBV-negative) seeking fertility assistance from January 2008 to December 2009 at the Womens Hospital in the School of Medicine at Zhejiang University were analysed for semen parameters. Couples in which the men were hepatitis B surface antigen (HBsAg)-seropositive were categorized as HBV-positive and included 587 in vitro fertilisation (IVF) and 325 intracytoplasmic sperm injection (ICSI) cycles from January 2004 to December 2009; negative controls were matched for female age, date of ova retrieval, ART approach used (IVF or ICSI) and randomized in a ratio of 1:1 according to the ART treatment cycles (587 for IVF and 325 for ICSI). HBV-infected men exhibited lower semen volume, lower total sperm count as well as poor sperm motility and morphology (P<0.05) when compared to control individuals. Rates of two-pronuclear (2PN) fertilisation, high-grade embryo acquisition, implantation and clinical pregnancy were also lower among HBV-positive patients compared to those of HBV-negative patients after ICSI and embryo transfer (P<0.05); IVF outcomes were similar between the two groups (P>0.05). Logistic regression analysis showed that HBV infection independently contributed to increased rates of asthenozoospermia and oligozoospermia/azoospermia (P<0.05) as well as decreased rates of implantation and clinical pregnancy in ICSI cycles (P<0.05). Our results suggest that HBV infection in men is associated with poor sperm quality and worse ICSI and embryo transfer outcomes but does not affect the outcome of IVF and embryo transfer. © 2011 AJA, SIMM &SJTU. All rights reserved. Source
Lu Y.-C.,Zhejiang University |
Ding G.-L.,Zhejiang University |
Yang J.,Zhejiang University |
Zhang Y.-L.,Zhejiang University |
And 9 more authors.
Human Reproduction | Year: 2012
Background The present study was designed to investigate the expression of small-conductance calcium-activated K+ channels 3 (SK3) in preimplantation embryos and to explore their role in the underlying mechanism of blastocyst hatching. Methods Human preimplantation embryos were donated by patients who achieved successful pregnancy with in vitro fertilization. Mouse preimplantation embryos in different stages were collected and cultured with or without siRNA cell injection. The expression of SK3 was examined by RTPCR, quantitative real-time PCR, western blot and immunofluorescence. Functional expression of SK3 was investigated using the patch-clamp technique. [Ca 2]i was measured by fluorescent imaging. Embryos were cultured in vitro to investigate the effect of SK3 knockdown or apamin, an SK3 inhibitor, on blastocyst hatching and F-actin formation. Results In human blastocysts, the level of SK3 expression was significantly lower in blastocysts that failed to hatch than in blastocysts that hatched successfully. In mouse embryos, SK3 mRNA and protein were not found in zygotes, but were detected from the 2-cell stage onward, with the highest levels observed in blastocysts. SK3 was predominately located in the trophectoderm cell membrane of expanded blastocysts. SK3 knockdown in trophectoderm cells not only suppressed the SK3 current, but also reduced [Ca2]i elevation and membrane potential hyperpolarization induced by thapsigargin. Although the formation of expanded blastocysts was not affected, blastocyst hatching and F-actin formation were significantly inhibited after SK3 knockdown in trophectoderm cells. Conclusions SK3-mediated [Ca 2]i elevation and membrane potential hyperpolarization in trophectoderm cells are important for blastocyst hatching, and defects in SK3 expression may contribute to infertility. © 2012 The Autho. Source
Zhu Q.,Zhejiang University |
Jin Y.,Zhejiang University |
Wang P.,Zhejiang University |
Wang H.,Zhejiang University |
And 5 more authors.
Cell Biology International | Year: 2015
The aims of this study were to delineate the expression of fatty-acid binding protein (FABP) 4 in human uterine endometrium and its function in the regulation of proliferation, migration and invasion of epithelial cells. Immunohistochenistry, immunofluorence and Western blotting were used to determine the expression and cellular localization of FABP4 in endometrium and endometrial epithelial cell lines. Interference of small ribonuclear acid (siRNA) and specific FABP4 inhibitor were used to inhibit FABP4. The proliferation, migration and invasion of epithelial cells were evaluated with CCK-8 assay, wound-healing test and transwell analysis respectively. We found that FABP4 was expressed by epithelial cells of proliferative endometrium and epithelial and stromal cells of secrectory endometrium. Epithelial cell lines Ishikawa and RL-952 expressed FABP4 and this expression was decreased by FABP4 siRNA. FABP4 siRNA and specific FABP4 inhibition significantly decreased the proliferation, migration and invasion of epithelial cell lines. We concluded that FABP4 is functionally expressed in endometrial epithelium and is necessary for maintaining the cell function of epithelial cells of endometrium. © 2015 International Federation for Cell Biology. Source
Feng G.-F.,Zhejiang University |
Zhang J.,Zhejiang University |
Feng L.-M.,Zhejiang University |
Shen N.-X.,Zhejiang University |
And 2 more authors.
Asian Journal of Andrology | Year: 2013
In this study, we aimed to determine whether the main mitochondrial DNA (mtDNA) haplogroups of the Han people have an impact on spermatozoa motility. We recruited 312 men who were consecutively admitted to two affiliated hospitals of College of Medicine, Zhejiang University from May 2011 to April 2012 as part of fertility investigations. Semen and whole blood samples were collected from the men. We determined the mtDNA haplogroups by analysing the sequences of mtDNA hypervariable segment I and testing diagnostic polymorphisms in the mtDNA coding region with DNA probes. No significant differences were found in the clinical characteristics of the mtDNA haplogroup R and non-R (P>0.05). Our results suggest that mtDNA haplogroup R is a strong independent predictor of sperm motility in the Han population, conferring a 2.97-fold (95%confidence interval: 1.74-4.48, P<0.001) decreased chance of asthenozoospermia compared with those without haplogroup R. © 2013 AJA, SIMM & SJTU. All rights reserved. Source
Qu F.,Zhejiang University |
Wang F.-F.,Zhejiang University |
Lu X.-E.,Zhejiang University |
Dong M.-Y.,Zhejiang University |
And 7 more authors.
Human Reproduction | Year: 2010
Background The present study was designed to evaluate whether the alteration of aquaporin-9 (AQP-9) expression in granulosa cells (GCs) of patients with polycystic ovary syndrome (PCOS) was associated with the hyperandrogenism in follicular fluid (FF). Methods We recruited infertile women with PCOS (n = 14) and infertile women with tubal blockage (controls, n = 31) for this study. We examined total testosterone (TT), free androgen index (FAI), sex hormone-binding globulin (SHBG), FSH, LH and estradiol in FF. Real-time PCR and western blotting were performed to assess AQP-9 expression in GCs, including effects of dihydrotestosterone (DHT) in vitro. Result SAQP-9 protein was localized in the nucleus, cytoplasm and cell membrane of the human GCs. The TT, FAI and LH levels were all higher, and SHBG levels lower, in the FF of women with PCOS versus controls (P = 0.0145, 0.0001, 0.0191, 0.0001, respectively). AQP-9 mRNA level in GCs of patients with PCOS was tightly correlated with the TT, SHBG levels and FAI in FF (P = 0.0020, 0.0001, 0.0020, respectively). In vitro, DHT (10-9 mol/l) decreased AQP-9 mRNA (lowest at 12 h) and protein levels in control GCs (P = 0.0005, 0.0247, respectively). The inhibitory effect of DHT on AQP-9 mRNA was attenuated by LY294002, a phosphatidylinositol 3-kinase (PI3K) inhibitor (P = 0.0013). Fifty micromolar 4-(hydroxymercuri) benzoic acid sodium salt (PMB) and 10-9 mol/l DHT blunted the swelling of GCs in hypotonic medium, respectively (P = 0.0350, 0.0027). CONCLUSION Hyperandrogenism in FF of women with PCOS inhibited AQP-9 in GCs through the PI3K pathway. © The Author 2010. Source