1.Progress in Research of the Role of Hepatitis C Virus Protein in Cell Signal Transduction Pathway
Mei XUE ; Jiejie DAI ; Xueshan XIA
Acta Laboratorium Animalis Scientia Sinica 2009;17(6):470-474
Abnormal cell signal transduction is associated with the occurrence and development of human diseases. Some virus pathogenicity and infection mechanism are due to virus antigen protein acting on the host cell signal transduction pathway, leading to host cell signal transduction disorder. Hepatitis C virus (HCV) is a major pathogen of chronic hepatitis C, which causes cirrhosis and hepatocellular carcinoma. But the pathogenesis of HCV and persistent infection mechanism remain far from clear. HCV pathogenesis may be related to the HCV protein expression interfering host cell signal transduction pathways. The studies of hepatitis C virus proteins acting on host cell signal transduction pathways, not only help to clarify the impact of its pathogenic mechanisms, but also benefit to new drug design and development for new treatment methods. This article summarizes the recent progress in research on the effect of hepatitis C virus protein in cell signal transduction pathways in the past few years.
2.Study on human immunodeficiency virus-1 super-infection among high-risk populations in Myanmar
Xiaojie LI ; Xueshan XIA ; Qianqiu WANG ; Yutaka TAKEBE ; Shigeru KUSAGAWA
Chinese Journal of Infectious Diseases 2011;29(4):239-241
Objective To investigate the occurrence and the characteristics of human immunodeficiency virus (HIV)-1 co/super-infection among high-risk populations in Myanmar.Methods Forty-six HIV-1 positive plasma in Myanmar were collected. Possible cases with HIV-1 co/super infection were identified by discordant sequence results obtained with different polymerase chain reaction (PCR)/sequencing primers or by ambiguous readings in direct sequencing. HIV-1 quasispecies in plasma were then characterized by clonal sequence analysis of independent PCR-clones generated by TA cloning method. Thereafter, their phylogeny and recombinant structure were investigated. Results Co/super infection was identified in 3 (6.5 %) cases among the 46 screened HIV-1 positive patients.All of these three patients were heterosexuals and were co/super infected with CRF01_AE/subtype B′recombinants. Conclusions HIV-1 co/super infections are relatively common and provide a prerequisite for rapid generation of new recombinant forms in Myanmar.
3.HIV and HCV: from Co-infection to Epidemiology, Transmission,Pathogenesis, and Treatment
Lei KANG ; Jing HU ; Xueshan XIA ; Jianguo WU
Virologica Sinica 2007;22(6):443-450
Human immunodeficiency virus (HIV) is the infectious agent causing acquired immunodeficiency syndrome (AIDS), a deadliest scourge of human society. Hepatitis C virus (HCV) is a major causative agent of chronic liver disease and infects an estimated 170 million people worldwide,resulting in a serious public health burden. Due to shared routes of transmission, co-infection with HIV and HCV has become common among individuals who had high risks of blood exposures. Among hemophiliacs the co-infection rate accounts for 85%; while among injection drug users (IDU) the rate can be as high as 90%. HIV can accelerate the progression of HCV-related liver disease, particularly when immunodeficiency has developed. Although the effect of HCV on HIV infection is controversial,most studies showed an increase in mortality due to liver disease. HCV may act as a direct cofactor to fasten the progression of AIDS and decrease the tolerance of highly active antiretroviral therapy(HARRT). Conversely, HAART-related hepatotoxicity may enhance the progression of liver fibrosis.Due to above complications, co-infection with HCV and HIV-1 has imposed a critical challenge in the management of these patients. In this review, we focus on the epidemiology and transmission of HIV and HCV, the impact of the two viruses on each other, and their treatment.
4.Prokaryotic expression of Hepatitis C Virus (HCV) NS3 protein and preparation of polyclonal antibody.
Tao SUN ; Guangwen YANG ; Jinyang ZHANG ; Xueshan XIA
Chinese Journal of Biotechnology 2015;31(5):711-721
To increase detection sensitivity and specificity on hepatitis C virus (HCV) is vital for prevention and controlling of the disease. To establish a more reliable detection method for HCV diagnosis, the full gene fragment of ns3 (non-structural protein of HCV) from recombinant plasmid of J6/JFH1 2a was amplified and then connected into the pET-28a prokaryotic expression vector, and the latter was subsequently transformed into Escherichia coli BL21 (DE3) to have the target protein expression. As a result, a protein with a molecular weight of 72 kDa was obtained and visualized in 10% SDS-PAGE. The purified NS3 protein was used as immunogen to inoculate BALB/c mice and the sera was collected after the fourth immunization. The antibody titer of serum is determined to be about 1:256000 with ELISA. Western blotting and indirect immunofluorescence analysis showed that the mouse polyclonal antibody could react specifically with the native NS3 protein in Huh 7.5.1 cells infected with HCV. These findings may provide basis for further preparation of monoclonal antibodies against NS3 and the development of related detection kit.
Animals
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Antibodies, Viral
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immunology
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Enzyme-Linked Immunosorbent Assay
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Escherichia coli
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Hepacivirus
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Mice
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Mice, Inbred BALB C
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Plasmids
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Viral Nonstructural Proteins
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biosynthesis
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immunology
5.Establishment of a EV71 virus infection model of tree shrew primary renal cells
Ming YANG ; Xiaoxing HU ; Wenguang WANG ; Li ZHANG ; Shuwei DONG ; Yue FENG ; Jiejie DAI ; Xueshan XIA
Acta Laboratorium Animalis Scientia Sinica 2017;25(2):117-122
Objective To establish an enterovirus 71(EV71) infection model of tree shrew primary renal cells.Methods Tree shrew primary renal cells were obtained by trypsin digestion.After subculture and purification,EV71 virus was used to infect these primary cells.The culture supernatant of these EV71-infected cells was collected for virus titer detection at 1,2,4,6 and 8 days post-infection.The cells were collected for detection of EV71 VP1 protein by Western blot assay.Furthermore,the expression and location of VP1 protein in the infected cells were detected by indirect immunofluorescence assay.Vero cells were taken as positive control to evaluate the infectivity of EV71 virus to tree shrew primary renal cells.Results Morphologically,the cultured cells were proved to be majorly consisted of the primary renal cells after subculture and purification.The obtained primary cells were infected by EV71 virus.The virus titer was up to 1.3×106 TCID 50/mL during 48-96 h post-infection,proving that EV71 virus infected and proliferated in the tree shrew primary renal cells.Western blot showed that the viral VP1 protein was detected from infected primary cells at 2 to 8 d post infection.VP1 protein was also observed in the cytoplasm at 2 to 6 d post infection by indirect immunofluorescence.Compared with Vero cells,the infectivity of EV71 virus to tree shrew primary renal cells and its proliferation were confirmed.Conclusions Based on the successful establishment of cell culture of tree shrew primary renal cells,the infectivity to the obtained cells and proliferation of EV71 virus in the cells are confirmed.The model of EV71 virus-infected tree shrew primary renal cells is initially established.
6.Progress in application of tree shrew models in research on human viral diseases
Anguo YIN ; Dexuan KUANG ; Xiaofei LI ; Yuan ZHANG ; Xiaomei SUN ; Xueshan XIA ; Jiejie DAI
Acta Laboratorium Animalis Scientia Sinica 2014;(2):86-89
Tree shrews get more and more concerns due to many of its physiological , biochemical and anatomical characteristics similar to those of human beings .Therefore, tree shrews models of human diseases such as viral diseases , neurological diseases and tumors attract more and more attention of researchers .In this article we will review the recent ad-vances in application of tree shrew models in research on human viral diseases .
7.Research progress of tree shrew models of viral hepatitis and modeling strategy
Yiquan PANG ; Yue FENG ; Xiaomei SUN ; Li LIU ; Jiejie DAI ; Xueshan XIA
Acta Laboratorium Animalis Scientia Sinica 2014;(2):95-110
Viral hepatitis is a major liver disease caused by virus infection .Viral hepatitis is popular in China , mainly caused by hepatitis B and hepatitis C viruses .Experimental animal model is a necessary platform for the research on mechanism of viral infection and pathogenicity , for treatment and vaccine development .Up to date, a great progress in the development of viral hepatitis animal models has been achieved in spite of the most of findings are limited to hepatitis B and C.Here, we summarize the recent findings of viral hepatitis animal models , focusing on the tree shrew animal model and its modeling strategy .
8.Establishment of a model of tree shrew primary small intestinal epithelial cells infected with human rotavirus G1P[8]
Daoqun LI ; Jie PENG ; Ziqin DIAN ; Wenguang WANG ; Amei ZHANGA ; Yue FENG ; Hua NIU ; Jiejie DAI ; Xueshan XIA
Acta Laboratorium Animalis Scientia Sinica 2017;25(2):111-116
Objective To explore the proliferation characteristics of primary small intestinal epithelial cells of tree shrews and the characteristics of human rotavirus(RV) G1P[8] infection to these cells,and establish a model of tree shrew primary small intestinal epithelial cells infected with human rotavirus G1P[8].Methods The primary small intestinal epithelial cells were obtained by collagenase Ⅺ and dispase I digestion from tree shrew.After purification and identification,the obtained primary small intestinal epithelial cells were infected with RV.Then,culture supernatants of infected cells were collected every 12 hours after infection.Viral titer and viral load were subsequently determined.Western blot and indirect immunofluorescence observation were used to detect the expression of RV protein VP6 in the primary cells.The infectivity of RV to the tree shrew primary cells was finally evaluated.Results After purification and identification of primary epithelial cells from the tree shrew,high purity above 90% primary tree shrew small intestinal epithelial cells was obtained.These primary small intestinal epithelial cells could be infected with RV virus by comparing the virus infectivity to primary renal cells,HCT116 cells and MA104 cells.The virus titer reached to 2.0×105TCID 50/mL at 72 h after infection.Using Western blot and indirect immunofluorescence observation,the specific viral protein of VP6 was determined to be expressed in the tree shrew primary small intestinal epithelial cells,and were located in the cytoplasm from days 1 to 5.Conclusions The separation,purification and cultivation methods of tree shrew primary small intestinal epithelial cells are successful,and the tree shrew model of RV-infected the tree shrew primary small intestinal epithelial cells is successfully established.
9.Analysis of National Natural Science Foundation of China during the period of " 12th Five-Year Plan" in Xinjiang Medical University
Xueshan CHEN ; Xia DUAN ; Apaer DILIBAIER ; Yimiti DELIXIATI
Chinese Journal of Medical Science Research Management 2018;31(3):189-194
Objective Information of the funding projects supported by the National Natural Science Foundation of Xinjiang Medical University from 2011 to 2015 was collected and analyzed in this paper.It also summarized the successful experiences,existing problems of project management during the "12th Five-Year" period for future improvement.Methods Statistical analysis was carried out on the funding projects by the National Natural Science Foundation of China from 2011 to 2015.Results With the support of the National Natural Science Foundation of China,the scientific research work experienced rapid progress and great improvement in Xinjiang Medical university.Conclusions By summarizing and improving the level of fund management in our university,we can also provide support for the sustainable development of our university's scientific research.
10.Screening of hepatocyte proteins interacting with hepatitis B virus X protein using CytoTrap yeast two-hybrid technique.
Baihai JIAO ; Yan WEN ; Xiaojia LIU ; Yue FENG ; Amei ZHANG ; Li LIU ; Xueshan XIA
Journal of Southern Medical University 2013;33(4):486-490
OBJECTIVETo screen the hepatocyte proteins that interact with hepatitis B virus X protein (HBx).
METHODSThe recombinant plasmid pSos-HBx was constructed by inserting Sos-HBx fragment into the bait vector, and after sequence verification the plasmid was transformed into competent yeast cells. The expression and self-activation of Sos-HBx protein was detected in the yeast cells. The hepatocyte proteins interacting with the bait protein was screened with CytoTrap yeast two-hybrid technique.
RESULTSThe reconstructed plasmid harboring HBx gene expressed Sos-HBx protein in the yeast cells without self-activation of the protein. CytoTrap yeast two-hybrid system identified 6 hepatocyte proteins that interacted with HBx, including fibronectin 1, translationally controlled tumor protein, IQ motif and WD repeats 1, follistatin, orosomucoid 1, and disulfide isomerase family A member 3.
CONCLUSIONSix HBx-binding hepatocyte proteins have been identified using the CytoTrap yeast two-hybrid system, which provides clues for further investigation of the role of HBx protein in hepatitis and liver cancer.
Genetic Vectors ; Hepatocytes ; metabolism ; Humans ; Plasmids ; Protein Interaction Domains and Motifs ; Proteins ; metabolism ; Trans-Activators ; metabolism ; Two-Hybrid System Techniques