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Cao X.,IVF Institute | Cao X.,Shanghai JiaoTong University | Liu Y.,Shanghai JiaoTong University | Chen G.,IVF Institute | Ding Z.,Shanghai JiaoTong University
Chinese Journal of Andrology | Year: 2010

Objective: To compare the efficiency and feasibility of human sperm acrosome reaction (AR) induced respectively by human zona pellucida glycoproteins, progesterone and calcium ionophore A23187 in vitro. Methods: Human spermatozoa were obtained using the swim-up method and re-suspended in Tyrode's buffer. Then the sperm AR were induced with human zona pellucida (ZP) glycoproteins, progesterone and calcium ionophore A23187 respectively. Sperm acrosomal status was assessed by Coomassie blue G250 staining and the percentage of acrosome-reacted cells was calculated in every sperm sample. Results The AR ratios of three groups under ZP, progesterone and A23187 treatments were (32.45±6.92)% (n=l 1), (42.41±19.19)% (n=22), (68.24 ± 8.85)%(n=43) respectively, and statistically significantly differences were observed between every two groups (P<0.05). Conclusion: The sperm AR ratios were relatively dispersive under induced by human ZP, progesterone and calcium ionophore A23187 respectively. The spermatozoa treated with A23187 showed the highest AR ratio and lowest individual variation among the sperm samples, however, the highest individual variations among the sperm samples was found in spermatozoa treated with progesterone.

Li Y.,Fudan University | Gong W.,Fudan University | Ma X.,Fudan University | Sun X.,IVF Institute | And 3 more authors.
Molecular Medicine Reports | Year: 2015

Epithelial ovarian carcinoma (EOC) is a lethal gynecological malignancy. Epithelial.mesenchymal transition (EMT) has an important role in the tumorigenesis and progression of EOC. During the process of EMT, the transforming growth factor-β (TGF-β)-Smad signaling pathway has been indicated to regulate cell motility and tumor development. Among numerous EMT-associated transcripts, Smad7 is considered to be an inhibitor, however its involvement together with TGF-β1 in the progression of ovarian cancer remains to be elucidated. The present study demonstrated that Smad7 was overexpressed in SK-OV-3 and stem-like side populations of EOC cells, both of which grow in an epithelial pattern. The transformation of cells from an epithelial to a mesenchymal phenotype was stimulated by TGF-β1 with a corresponding increase in Smad7 expression in SK.OV.3 cells. These results indicate that Smad7 is a regulator in the maintenance of the epithelial phenotype in EOC cells, and may serve as an inhibitory element which targets TGF-β-stimulated EMT. Furthermore, inhibition of Smad7 resulted in cellular mesenchymal transformation, with an increased expression of N-cadherin and a decreased expression of E-cadherin. The invasiveness and migratory capabilities of Smad7 small hairpin RNA transduced EOC cells was also reduced. The findings of the present study have identified Smad7 as a fundamental factor in the maintenance of epithelial growth of EOC cells. Reversal of EMT results in a mesenchymal-epithelial transition, which is necessary for EOC cell colonization at metastatic sites.

Novik V.,IVF Institute | Moulton E.B.,IVF Institute | Sisson M.E.,IVF Institute | Shrestha S.L.,IVF Institute | And 4 more authors.
Molecular Cytogenetics | Year: 2014

Background: Most previous studies of chromosomal mosaicism in IVF embryos were performed by fluorescence in situ hybridization (FISH) methods. While there are reports implicating chromosome aneuploidy in implantation failure following transfer and pregnancy loss by spontaneous miscarriage, the significance of mosaicism for the developmental potential of growing embryos is unknown. However, the low prevalence of chromosomal mosaicism in chorionic villus sampling and amniotic fluid specimens suggests the presence of selection against mosaic embryos for implantation and early pregnancy. The absence of evidence for selective allocation of abnormal cells to the trophectoderm (TE) of mosaic blastocysts permits these cells to be a good proxy for embryonic mosaicism detection by chromosomal microarrays (CMA). The purpose of this study was to establish the limits of detection and the prevalence of chromosome mosaicism in day 5/6 human embryos using CMA with TE biopsies. Results: From reconstitution experiments we established log2 ratio thresholds for mosaicism detection. These studies indicated that chromosomal mosaicism at levels as low as between 25-37% can be consistently identified. Follow-up studies by FISH on non-transferred abnormal embryos confirmed the diagnostic accuracy of CMA testing. The number of cells in a TE biopsy can influence mosaicism detection. Conclusions: Chromosomal microarrays can detect mosaicism in TE biopsies when present at levels as low as between 25-37% and the prevalence of day 5/6 blastocysts which were mosaic and had no other abnormalities reached 15% among a cohort of 551 embryos examined. Validated protocols for establishing detection thresholds for mosaicism are important to reduce the likelihood of transferring abnormal embryos. © 2014 Novik et al.; licensee BioMed Central Ltd.

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