Yi-Zhan Cao

Fourth Military Medical University, Xi’an, Liaoning, China

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Publications (6)2.55 Total impact

  • Yi-zhan Cao, Qing-he Nie
    Zhongguo wei zhong bing ji jiu yi xue = Chinese critical care medicine = Zhongguo weizhongbing jijiuyixue 08/2003; 15(7):441-4.
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    ABSTRACT: To construct three recombinant shuttle plasmids of adenovirus expression vector which can express hepatitis C virus(HCV) different structure genes(C, C+E1, C+E1+E2) in order to pack adenovirus expression vectors which can express HCV different structure gene effectively. The different HCV structure genes derived from the plasmid pBRTM/HCV1-3011 by using polymerase chain reaction (PCR) were inserted into the backward position of cytomegalovirus(CMV) immediate early promotor element of shuttle plasmid(pAd.CMV-Link.1) of adenovirus expression vector respectively, then the three recombinant plasmids (pAd.HCV-C, pAd.HCV-CE1, pAd.HCV-S) were obtained. The recombinant plasmids were identified by endonuclease, PCR and sequencing. HCV structure genes were expressed transiently with Lipofectamine 2000 coated in HepG2 cells which were confirmed by immunofluorescence and Western-Blot. Insert DNAs of the three recombinant plasmids' were confirmed to be HCV different structure genes by endonuclease, PCR and sequencing. The three recombinant plasmids can express HCV structure gene (C, C+E1, C+E1+E2) transiently in HepG2 cells which were confirmed by immunofluorescence and Western-Blot. The three recombinant shuttle plasmids of adenovirus expression vector can express HCV structure gene(C, C+E1, C+E1+E2) transiently. This should be useful to pack adenovirus expression vector which can express HCV structure genes.
    Zhongguo wei zhong bing ji jiu yi xue = Chinese critical care medicine = Zhongguo weizhongbing jijiuyixue 03/2003; 15(2):69-72.
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    ABSTRACT: Objective. To investigate the rule of intensity and duration of HSP70 expression in rat brain and its relationship with brain injury after repeated +Gz exposures. Method. SD male rats were arranged into control group, +2 Gz, +4 Gz, +6 Gz, and +10 Gz exposure groups. Rat brains were taken 6 h, 10 h, 1 d, 2 d, 4 d or 6 d after +Gz exposure for histopathologic and immunohistochemic or in situ hybridization studies. The expression of HSP70 and HSP70 mRNA or morphology of neurons were observed. Result. The intensity and duration of HSP70 expression were weak and brief at +2 Gz exposure, but was relatively extensive. There was a middling reaction of HSP70 only in hippocampal area after +10 Gz exposure. The duration, extension and intensity of HSP70 expression were wide, long and strong after +4 Gz and +6 Gz exposures. After 1 or 3-5 times exposures, the HSP70 expression reached its peak on the first day after +4 Gz exposures, and dropped obviously on the second day. However the expression of HSP70 maintained a high level after 2 d and was still higher than normal on the 6 d after 3-5 times repeated +4 Gz exposures. The distribution of HSP70 mRNA expression was as same as that of the HSP70 expression but the peak appeared much earlier (10 h) and its duration was shorter. After +10 Gz/5 min exposure, degenerated neurons were found in cortex, hippocampus and thalamus regions while the number of degenerated neurons were obviously decreased in such areas in pre-exposure groups with repeated +4 Gz/3 min for 3-5 times. Conclusion. The intensity and duration of HSP70 and HSP70 mRNA expression after +4 Gz and +6 Gz exposure were stronger and longer than +2 Gz and +10 Gz exposure. The degree of neuron damage after +10 Gz/5 min exposure in pre-exposure groups with repeated +4 Gz/3 min 3-5 times was obviously slight comparing with that of single +10 Gz exposure group.
    Hang tian yi xue yu yi xue gong cheng = Space medicine & medical engineering 01/2003; 15(6):391-6.
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    ABSTRACT: To set up a new method to detect tissue inhibitors of metalloproteinase-1 and -2(TIMP-1 and TIMP-2) in sera of patients with hepatic cirrhosis, and to investigate the expression and location of TIMP-1 and TIMP-2 in liver tissue of patients with hepatic cirrhosis, and the correlation between TIMPs in liver and those in sera so as to discuss whether TIMPs can be used as a diagnosis index of hepatic fibrosis. The monoclonal antibodies (McAbs) of TIMP-1 and TIMP-2 were used to sensitize erythrocytes, and solid-phase absorption to sensitized erythrocytes (SPASE) was used to detect TIMP-1 and TIMP-2 in the sera of patients with hepatic cirrhosis. Meanwhile, with the method of in situ hybridization and immunohistochemistry, we studied the mRNA expression and antigen location of TIMP-1 and TIMP-2 in the livers of 40 hepatic cirrhosis patients with pathologic diagnosis. With SPASE, they were 16.4% higher in the acute hepatitis group, 33.3% higher in the chronic hepatitis group, and the positive rates were 73.6% and 61.2% respectively in sera of hepatic cirrhosis patients, which were remarkably higher than those in chronic hepatitis and acute hepatitis group (P<0.001). In 40 samples of hepatic cirrhosis tissues, all of them showed positive expression of TIMP-1 and TIMP-2 mRNA detected with immunohistochemistry or in situ hybridization (positive rate was 100%). Expression of TIMPs in different degrees could be found in liver tissue with cirrhosis. TIMPs were located in cytoplasm of liver cells of patients with hepatic cirrhosis. There was a significant correlation between serum TIMPs level and liver TIMPs level. SPASE is a useful method to detect the TIMP-1 and TIMP-2 in sera of patients with hepatic cirrhosis, and TIMP-1 and TIMP-2 can be considered as a useful diagnostic index of hepatic fibrosis, especially TIMP-1.
    World Journal of Gastroenterology 04/2002; 8(2):282-7. · 2.55 Impact Factor
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    ABSTRACT: AIM:To construct a replication-deficient recombinant adenovirus expression vector of HCV C. METHODS:The HCV core gene was cloned at the down- stream of CMV promoter of the adenoviral shuttle plasmid pAd.CMV-link.1, and the resultant recombinant plasmid pAd. HCV-C was cotransfected into 293 cell together with plasmid pJM17 containing adenoviral genome, then the adenovirus expression vector was obtained, and identified by infecting test,electronic microscope observation and PCR co-amplification. The plasmid pAd.HCV-C was identified by endonuclease, PCR and sequencing.The expressive activity of adenovirus vector was identified by immunofluorescence and Western blot. RESULTS:HCV core gene in the inserted DNA of pAd.HCV-C was confirmed by endonuclease, PCR and sequencing. Results of infecting test, electronic microscopic observation and PCR co-amplification showed that the adenovirus vector had been constructed successfully. Expression of HCV core antigen was proved in the HepG2 cells by immunofluorescence and Western blot.
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    ABSTRACT: AIM:To evaluate the effect of hammerhead ribozyme 213 (Rz 213) against hepatitis C virus (HCV) infection. METHODS:Rz213 cleaving 5'noncoding region (5'NCR) of HCV was beforehand transfected in a human hepatic carcinoma cell (HHCC) line and selected for G418 resistance. Cells stably expressing Rz213 were retransfected with pCMVNCRluc containing 5'NCR-luc fusion genes by lipofectAMINE;luciferase activity in lysate of transfactant was measured in scintillation counter. RESULTS:HHCC cells stably expressing Rz213 exhibited significant resistance to retransfection of targeting gene.