4.Progress in the studies of prostate cancer related molecules.
Wei SHI ; Li DONG ; Jun-sheng BAO
National Journal of Andrology 2015;21(4):357-362
Prostate cancer is one of the common malignant tumors of the urinary system and mostly found in elderly men. Like most tumors, prostate cancer involves a variety of molecules in its occurrence and progression. More studies on the development of prostate cancer focus on the tumor markers, DNA damage repair related genes, and tumor invasion and metastasis related factors. This article presents an overview on the research progress in these three aspects.
Biomarkers, Tumor
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Biomedical Research
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DNA Repair
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Disease Progression
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Humans
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Male
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Neoplasm Invasiveness
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Prostatic Neoplasms
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genetics
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pathology
5.Synthesis of DNA Encoding Botulinum Neurotoxin Receptor syt II and Its Fusion Expression in E.coli
Jing SHI ; Hui WANG ; Shi-Zhong BAO ; Xiao-Jun HOU ; Jun YIN ;
China Biotechnology 2006;0(07):-
In order to construct a prokaryotic expression vector of human receptor syt II N-fragment and to express recombinant MBP-Syt fusion protein in E.coli and to purify and identify its activity. According to codon preference of E.coli, a DNA fragment encoding human syt II N-fragment was synthesized, and then cloned into prokaryotic vector pMAL-c2x for sequencing. Then the recombinant plasmid pMAL-Syt was introduced into E.coli ER2566 by transformation for expression and the obtained engineered bacteria were induced by IPTG. The fusion protein was purified by amylose resin affinity chromatography and identified by SDS-PAGE and Western blot. The binding activity of the protein was determined by ELISA. It is concluded that MBP-Syt protein is of good binding activity.
6.mRNA expression change of myelin gene in hippocampus following cerebral ischemia
Ying-Zhu CHEN ; Huan BAO ; Ye TIAN ; Shi-Yao BAO ; Jun XU ; Cheng-Lin YUAN ;
Chinese Journal of Emergency Medicine 2006;0(09):-
Objective To study the mRNA expression of myelin basic protein (MBP) and myelin oligodendregha glyeoprotein (MOG) in hippocampus of rats following global brain ischemia.Method The four- vessel occlusion animal model in the Sprague-Dawley rats was used in this study.The mRNA expression levels of MBP and MOG in the hippocampus of rats were analyzed by reverse transcription polymerase chain reaction (RT- PCR) at day 2,4,7,14 and 28 days after global brain ischemia.There were eight rats at each time-point and sham operated group.Results The mRNA expression of both MBP and MOG in hippocampus of rats decreased at 2 days after global brain ischemia.The gene expression of myelin gene decreased significantly at 7 days and it reached to the lowest level at 28 days.Compared with sham operated group,the gene expression of MBP and MOG in hippocampus of rats decreased significantly at 7,14 and 28 days after global brain ischemia (P
7.The Recombinant Expression and Receptor-binding Activity of the B Subunit of Shiga-like Toxin Type Ⅱ
Shi-Zhong BAO ; Jing SHI ; Kun CAI ; Jun YIN ; Hui WANG ;
China Biotechnology 2006;0(10):-
Objective:To express the B subunit of Shiga-like toxin type Ⅱ,and analyze its expression form and receptor-binding activity.Methods:The slt2b gene was obtained from EHEC O157∶H7 by PCR,and cloned to the expression vector pET22b(+).The genetically engineered bacteria pET22b(+)-stx2B/BL21 expressed the recombinant StxB after induced with IPTG.The renatured inclusion bodies were purified by ion exchange chromatography.The expression form of rStx2B was investigated by denaturing and native electrophoresis.The receptor-binding activity was confirmed by fluorescence detection and flow cytometer.Result:The constructed genetically engineered bacteria expressed the rStx2B at a high level.The purified protein was obtained after denaturation,renaturation and ion exchange chromatography.According to the denaturing and native electrophoresis,the rStx2B was expressed in a dimmer form,which consists of two monomers cross linked with disulfide bridge.The rStx2B showed good receptor-binding activity by Hela-binding assay.Conclusion:The genetically engineered bacteria were constructed successfully.The receptor-binding activity of rStx2B was independent of the pentamers.
8.Construction and Screening of a Phage Display Library of Repertoire Single Chain Fv Antibody from Mouse Immunized with BoNTB/Hc
Xiu-Qing YANG ; Hui WANG ; Jing SHI ; Kun CAI ; Xiao-Jun HOU ; Shi-Zhong BAO ; Jun YIN ;
Microbiology 1992;0(06):-
To produce antibodies capable of neutralizing botulinum neurotoxin type B(BoNT/B),We cloned the carboxy-terminal end of Hc containing the major determinants responsible for specific toxin,induced and purifed.The heavy-chain and kappa light-chain variable region gene repertoire of immunoglobulin were amplified individually from the spleen cell mRNA by RT-PCR and joined as a single-chain Fv(scFv)DNA fragment.These fragment were cloned into the phagemid pCANTAB5E and the phage display library was constructed.Results showed that the high affinity scFv was obtained after 4 rounds of panning,with its DNA sequence conforming to that of mouse antibody.
9.Effect of different pressure oxygen pre-breathe in diving decompression sickness of rats.
Fang-fang WANG ; Yi-qun FANG ; Pu YOU ; Xiao-chen BAO ; Jun MA ; Shi ZHANG
Chinese Journal of Applied Physiology 2015;31(5):401-404
OBJECTIVETo investigate the effect of different pressure oxygen pre-breathing in preventing decompression sickness of rats.
METHODSForty male SD rats were randomly divided into 4 groups: decompression sickness (DCS) group and three oxygen pre-breathing groups with 1 ATA, 2 ATA and 3 ATA pressure respectively. The rats of DCS group were placed in the hyperbaric chamber and the chamber was compressed evenly within 3 minutes to depths of 7 absolute atmosphere(ATA) and held at the designated depth for 60 min, then decompressed (3 min) at constant speed to the surface pressure. After that, the rats were taken out for further detection. While the rats of oxygen pretreatment groups pre-breathed different pressure oxygen for 20 min before entering into chamber. The mortality and behavioral of rats were observed with 30 min post decompression. The dry/wet ratio of the lung, protein levels in the bronchoalveolar lavage fluid (BALF), and the inflammatory cytokine tumor necrosis factor (TNF-alpha) expression were also tested.
RESULTSCompared with that of the DCS group, the mortality and morbidity of oxygen pre-breathe groups didn't change obviously. But the total BALF protein level and the inflammatory cytokine TNF-alpha expression of 1 ATA oxygen pre-breathe group were obviously decreased, while the dry/wet ratio of lung as obviously increased instead (P < 0.05).
CONCLUSIONAlthough preoxygenation can' t obviously change the mortality and mobidity of rats, normal pressure oxygen pre-breathing can mitigate the protein infiltration in BALF and the expression of inflammatory cytokine in lung tissue.
Animals ; Bronchoalveolar Lavage Fluid ; chemistry ; Decompression Sickness ; Diving ; Lung ; pathology ; Oxygen ; physiology ; Pressure ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; metabolism