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Articles by S.S. Lu
Total Records ( 2 ) for S.S. Lu
  X.M. Song , S.S. Lu , M. Wang , Q.Y. Li , D. Li , X.G. Yang , Y.Q. Lu , M. Zhang and K.H. Lu
  This study evaluated the effects of Glutathione (GSH) supplemented in the semen extender during the buffalo sperm sorting process on sperm quality, embryonic development after IVF and pregnancy rate after AI. The percentage of sperm motility, plasma membrane integrity and DNA fragmentation were detected by flow cytometry or by microscopy in stained, sorted and frozen semen treated with or without 0.75 mM GSH during the flow sorting procedure. The cleavage and blastocyst rates were examined at day 2 and 6-8 after IVF with frozen semen treated with or without 0.75 mM GSH. Pregnancy diagnosis was determined by transrectal palpation at 90 day after AI with frozen semen treated with or without 0.75 mM GSH. The percentage of sperm with Progressive Motility (MP, %) was significantly higher (p<0.05) in sorted semen supplemented with 0.75 mM GSH than that in the control. The percentages of moribund, dead and Phosphatidylserine (PS) translocated sperm detected by flow cytometry were significantly decreased (p<0.05) in frozen semen supplemented with GSH compared to the control. Higher blastocyst and pregnancy rates (p<0.05) were found after IVF and AI with frozen sperm treated with 0.75 mM GSH than that in the control group. In conclusion, addition of 0.75 mM GSH to the semen extenders (stained, sorted and frozen) during the sperm sorting process can improve sperm quality in vitro embryonic development and in vivo fertility after AI thus indicating potential for commercial application in buffalo sperm sorting.
  X.B. Zheng , X.M. Cen , L.X. Wang , G.Y. Xin , X.H. Fu , P. Liu , G.H. Li , M. Zhang , S.S. Lu and K.H. Lu
  Nanog is one of important transcription factors to maintain characteristics of pluripotent stem cells. The present study was to clone Nanog of Capra hircus to express His-Nanog protein in E. coli BL 21 cells and further to purify it. The total RNA was extracted from primordial genital ridge tissues of a fetal lam and by means of RT-PCR, Nanog gene was amplified which was subcloned to pET32a to construct its prokaryotic expression vector. Confirmed by restrictive endonuclease digestion and DNA sequencing, the recombinant plasmid was transformed into E. coli BL21(DE3) and His-Nanog fusion protein was expressed by the induction of IPTG and identified with SDS-PAGE analysis. Under denaturing condition, the His-Nanog protein was purified by using Ni-NTA resin and verified by Western blotting assay. The results showed that The Open Reading Frame (ORF) of Nanog gene in Capra hircus is composed of 903 nucleutide acids, coding 320 amino acids; SDS-PAGE assay showed that His-Nanog fusion protein was efficiently expressed in form of inclusion bodies in E. coli BL21 (DE3) inclusion bodies were solubilized in 6 mol L-1 GuHCl, His-Nanog fusion protein with higher purity was purified by using Ni-NTA resin; Western blotting assay showed that the purified His-Nanog could bind to anti-His tag antibody specifically indicating the expected immunogenicity. This recombinant protein could be used directly to prepare polyclonal or monoclonal anti-Nanog antibody which will lead to study Nanog’s function or characteristics of pluripoten stem cells (such as iPS cells) in Capra hircus.
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