1.Construction,expression and purification of HIV-1 p24-gp41 fusion protein in E.coli
Weimin JIANG ; Hongzhou LU ; Xiaozhang PAN
Chinese Journal of Infectious Diseases 2001;0(03):-
Objective To construct recombinant plasmid with p24 and gp41 gene, express fusion protein in E.coli. Methods To design primer with restriction endonuclease position,and amplify p24 and gp41 by RT-PCR, link both into pMD18-T vector.To choose correct clone with target gene.Then p24 fragment will be cleaved and linked into pMD18-T vector within gp41 gene. Both post-linked gene will be cleaved and linked into pET21a vector. The vector will be transformed into E.coli. And protein is highly effective expressed in E coli. Western blotting proved that the expressed product could react with 6?his antibody. Result Fusion protein p24-gp41 is highly effective expressed in E.coli. Conclusion Fusion protein p24-gp41 is highly effective expressed in E.coli in pET21a vector.
2.Measurement of the CD4 and CD8 normal value range in adults from Shanghai area
Weimin JIANG ; Xiaozhang PAN ; Laiyi KANG
Chinese Journal of Infectious Diseases 1997;0(04):-
Objective To measure normal range of CD4 from Chinese adults in Shanghai area by flow cytometry. Methods CD4 and CD8 value were measured through flow cytometry in adult blood specimens from various age and gender group. Results There were no difference in CD4 value among different sex and age groups, with the mean value being 726.99?255.21. However, CD8 and CD4/CD8 ratio vary significantly among different groups. The mean value of CD8 was 539.58?134.07, while the mean value of CD4/CD8 1.49?0.57. Conclusions The mean value of CD4 in Shanghai adults is about 100/mm 3 lower than that in American.
3.Expression of viral infectivity factor protein of human immunodificiency viruses and its biological function
Yunzhi ZHANG ; Hong JIN ; Hongzhou LU ; Xunjia CHENG ; Xiaozhang PAN
Chinese Journal of Infectious Diseases 2008;26(4):231-234
Objectives To analyze the characteristic of HIV-1 viral infectivity factor (Vif) gene variants isolated from Shanghai. To construct the prokaryotic expression vector of HIV-1 vif gene and understand its immunogenieity. Methods HIV-1 vii genes were amplified and sequenced from 23 serum samples of HIV-1 infected patients in Shanghai and then compared with the international standard HIV-1 strain. Subsequently, these amplified Vif fragments were sub cloned into pETS2b(+)expression vector. The recombinant prokaryotie plasmids pETS2b (+)-HIV-1/Vif were then transferred into BL21DS(Star)cells for expressing and purifying HIV-1 Vif protein. HIV-1 Viff rat polyclonal antibody was then preparing by injecting the purified Vif proteins into the mice. ELISA was used to determine the purity of Vif proteins and the immunogenicity of its polyclonal antibodies. Results The nucleotide acid mutation rate of HIV-1 vif gene in Shanghai AIDS patient was (0. 179±0. 006)% compared with the international standard HIV-1 strain. Some similar mutations in vif gene were found in HIV-1 strains isolated from Shanghai while the amino acid sequence between 151 and 240 in Vif protein was conserved. The construction of HIV-1 Vif prokaryotic expression plamids and the preparation of Vif polyclonal antibodies were successfully done in this study. The reaction between recombinant HIV-1 Vif protein and the serum from HIV-1 infected patient was not significantly different from that between the recombinant protein and healthy control serum(P>0.05). HIV-1 Vif polyclonal antibody reacted differently with recombined HIV-1 Vif protein compared with healthy control serum samples (t=178.61, P<0.01). Conclusions The Vif gene mutation rate is high in HIV-1 strains isolated from Shanghai compared with international standard HIV-1 strain. The prokaryotic expression plasmids of HIV-1 vif antigen are successfully constructed and Vif polyclonal antibodies are prepared well.
4.CSF-VP shunt placement and albendazole therapy for cerebral cysticercosis.
Liping ZHU ; Xinhua WENG ; Yaozhong SHI ; Xiaozhang PAN ; Ling MO
Chinese Medical Journal 2002;115(6):936-938
Cerebral cysticercosis is the most common worldwide parasitic infection of the central nervous system. Intraventricular involvement is apparent in 15% to 28.8% of cases with neural compartment infestation.' Although different forms of the disease (parenchymatous, subarachnoid, and mixed form ) have been treated successfully with chemotherapy, direct surgical excision of simple cystic lesions appears to be an adequate primary therapeutic strategy in the majority of intraventricular forms. In recent years, however, some authors have advocated the use of anthelmintic treatment in all cases of intraventricular cysts so that surgical procedures of the posterior fossa and their potential complications can be avoided. The strict definition for managing the spectrum of intraventricular infestation remains controversial. We present our experience in the treatment of a patient with primary isolated intraventricular cysticercosis.
Albendazole
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therapeutic use
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Anthelmintics
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therapeutic use
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Child
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Female
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Humans
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Neurocysticercosis
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therapy
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Ventriculoperitoneal Shunt