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| Articles
by
S.S. Lu |
Total Records (
2 ) for
S.S. Lu |
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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
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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. |
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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
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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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