Po Tien

Wuhan University, Wu-han-shih, Hubei, China

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Publications (149)585.3 Total impact

  • [Show abstract] [Hide abstract]
    ABSTRACT: The interferon (IFN) immune system plays an essential role in protecting the host against most viral infections. In order to explore the interactions between the IFN pathway and Respiratory syncytial virus (RSV) infection, and to identify potential IFN-stimulated genes (ISGs) that may be involved in suppressing the replication of RSV, we utilized an IFN pathway-specific microarray to study the effects of RSV infection on the IFN pathway in HeLa cells. We showed that RSV infection enhanced the expression of a series of ISGs, including oligoadenylate synthetase 2 (OAS2), interferon-induced transmembrane protein 1 (IFITM1) and myxovirus-resistance 2 (Mx2). Our results also showed that the IFITM proteins potently inhibited RSV infection mainly by interfering with both virus entry and the subsequent replication steps, but not the attachment process. The anti-viral effect of IFITM3 protein was not affected by ubiquitination modification. Furthermore, knocking down the endogenous and IFN-induced expressions of IFITM1 and IFITM3 proteins facilitated RSV infection. Expression of the IFITM proteins was found to delay the phosphorylation of interferon regulatory factor-3 (IRF3) through interfering with the detection of viral RNA by the melanoma differentiation-associated gene 5 (MDA5) and the retinoic acid-inducible gene I (RIG-I). These results demonstrated that the restriction of RSV infection by the IFITM proteins was achieved through the inhibition of virus entry and replication, and they provide further insight for exploring the mechanism of IFITM proteins-mediated virus restriction.
    Journal of General Virology 09/2014; · 3.13 Impact Factor
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    ABSTRACT: Series of non-nucleoside reverse transcriptase inhibitors derived from indole-based α-amino acids were designed and synthesized. Their inhibitory activities have been detected by a TZM-bl cell assay on HIV virus type HIV-1IIIB. The comprehensive understanding of the SAR was obtained by utilizing the variation of the substituents of the indole-based α-amino acids. From the screened compounds, the novel inhibitors 19 and 29 were identified to be highly potent candidates with EC50 value of 0.060 μM and 0.045 μM respectively (CC50 value of 109.545 μM, 49.295 μM and SI value of 1825.8, 1095.4). While, in most cases, that variation of substituents at different position had a significant effect on the potency of activities. The results also indicate that the indole-based α-amino acids as efficient NNRTIs displayed comparable anti-HIV-1 activities with the reference drug NVP. We hope the identification of these indole-based amino acids as efficient NNRTIs of RT could stimulate researchers to develop more diversified anti-HIV drugs.
    Organic & Biomolecular Chemistry 08/2014; · 3.57 Impact Factor
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    ABSTRACT: Cytotoxic T lymphocyte (CTL) epitopes in the HBV protein of hepatitis B virus (HBV) may play a key role in viral control and liver damage. The aim of this study was to identify and study the function of HLA-A∗33:03-restricted CTL epitopes in HBV protein of the HBV genotypes B and C, which are epidemic in China. Sixteen HBV peptides were predicated by computational analysis, and synthesized peptides were examined for their affinity to HLA-A∗33:03 using a stable cell line. After being analyzed by enzyme-linked immunospot and cytolytic activity assays, as well as the tetramers staining method using peripheral blood mononuclear cells isolated from HBV-infected patients, five peptides (Hbs245–253, HBs335–343, HBc119–127, HBc104–112, and HBp391–399) were chosen to further confirm their HLA_A∗33:03 restriction in transgenic mice.
    Biochemical and Biophysical Research Communications 06/2014; 449(1):135–140. · 2.41 Impact Factor
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    ABSTRACT: Biomolecule-nanoparticle hybrid bioconjugates based on bioscaffolds such as protein cages and virus capsules have been widely studied. Highly stable and durable biotemplates are a vital pillar in constructing bio-inorganic functional hybrid composites. Here, we introduce a highly heat-resistant coat protein (CP) of Sulfolobus tengchongensis spindle-shaped virus 1 (STSV1) isolated from the hyperthermophilic archaeon as a prospective biological matrix. Our experiments showed that STSV1 CP was successfully cloned and solubly expressed in the Escherichia coli Rosetta-(DE3) host strain. Protein expression was verified by SDS-PAGE and western blot analysis of the reference C-terminally six-histidine (His6) tagged STSV1 CP (HT-CP). Thermal stability experiments showed that the STSV1 coat protein remained fairly stable at 80 °C. The proteins can be purified facilely by heat treatment followed by size exclusion chromatography (SEC). Transmission electron microscopy (TEM) analysis of the purified STSV1 CP protein aggregates demonstrated that the protein could self-assemble into rotavirus-like nanostructures devoid of genetic materials under our experimental conditions. Similar results were obtained for the HT-CP purified by heat treatment followed by Ni-NTA and SEC, indicating that moderately engineered STSV1 CP can retain its self-assembly property. In addition, the STSV1 CP has a high binding affinity for TiO2 nanoparticles. This illustrates that the STSV1 CP can be used as a bioscaffold in nanobiotechnological applications.
    Extremophiles : life under extreme conditions. 06/2014;
  • Zhuo Yang, Po Tien
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    ABSTRACT: MicroRNAs (miRNAs) play an important role in infection and replication of virus in host cells. To identify cellular miRNAs involved in the host response to enterovirus 71 (EV71) infection, we examined miRNAs effects on the replication of EV71 in rhabdomyosarcoma cells. We constructed target gene of miRNAs screening system. 3'untranslated region (UTR) dual luciferase reporter analysis was used to identify putative miRNA targets in the EV71 virus genome. First, 13 segments of EV71 virus genomes were inserted to the pMIR vector and the luciferase expression were assayed to identify the target gene of putative miRNA. The reporter gene expression of the cells transfected with the vector containing 5'-UTR was significantly downregulated. Then we screened the miRNAs that may target to 5'-UTR using online analysis programs. Furthermore, Western blotting and real-time PCR test were performed to investigate the effect of miRNAs on viral replication. The study showed that miR373 and miR542-5p could suppress the replication of EV71 virus through binding to the 5'-UTR gene. Cellular miRNAs could regulate the replication of EV71 virus in host cells, and our paper should report the role of miR373 and miR542-5p in this regulation for the first time. Our findings supported the notion that the cellular miRNAs might be essential in the host-pathogen interactions.
    06/2014; 30(6):943-53.
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    ABSTRACT: Cytotoxic T lymphocyte (CTL) epitopes in the HBV protein of hepatitis B virus (HBV) may play a key role in viral control and liver damage. The aim of this study was to identify and study the function of HLA-A∗33:03-restricted CTL epitopes in HBV protein of the HBV genotypes B and C, which are epidemic in China. Sixteen HBV peptides were predicated by computational analysis, and synthesized peptides were examined for their affinity to HLA-A∗33:03 using a stable cell line. After being analyzed by enzyme-linked immunospot and cytolytic activity assays, as well as the tetramers staining method using peripheral blood mononuclear cells isolated from HBV-infected patients, five peptides (Hbs245-253, HBs335-343, HBc119-127, HBc104-112, and HBp391-399) were chosen to further confirm their HLA_A∗33:03 restriction in transgenic mice.
    Biochemical and Biophysical Research Communications 05/2014; · 2.41 Impact Factor
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    ABSTRACT: To find novel compounds against H5N1, three series of known or novel small molecular polyphenols were synthesized and tested in vitro for anti-H5N1 activity. In addition, the preliminary structure-antiviral activity relationships were elaborated. The results showed that some small molecular polyphenols had better anti-H5N1 activity, and could serve as novel virus entry inhibitors against H5N1, likely targeting to HA2 protein. Noticeably, compound 4a showed the strongest activity against H5N1 among these compounds, and the molecular modeling analysis also suggested that this compound might target to HA2 protein. Therefore, compound 4a is well qualified to serve as a lead compound or scaffold for the further development of H5N1 entry inhibitor.
    Bioorganic & medicinal chemistry letters 04/2014; · 2.65 Impact Factor
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    ABSTRACT: Exposure of cells to type I interferon (IFN) induces an antiviral state that prevents viral infection, but viruses can utilize multiple tactics to antagonize the host immune system. Enterovirus 71 (EV71) and Coxsackievirus A16 (CVA16) are two major pathogens that cause hand, foot, and mouth disease (HFMD), which is prevalent among children. We found that both EV71 and CA16 have different reactions to type I IFN pretreatment and induction patterns of type I IFN on Rhabdomyosarcoma (RD) cells. Further, a human-α and β IFN PCR array was employed to analyze the expressions of 84 genes related to the type I IFN pathway. We found significant up-regulation of multiple genes in the presence of type I IFN and differential regulation patterns during EV71 or CA16 infection in RD cells. For instance, EV71 infection repressed the JAK-STAT signaling pathway and interferon-stimulated gene (ISG) expression, whereas CA16 infection normally triggers the JAK-STAT pathway, leading to the expression of ISGs. Taken together, this study provides a comprehensive view of the differential impacts of EV71 and CA16 infection on 84 genes in the IFN pathway, shedding light on the different resistances of these viruses to type I IFN treatment and cytotoxic effects in RD cells.
    Biochemical and Biophysical Research Communications 04/2014; · 2.41 Impact Factor
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    ABSTRACT: Biomolecule-mediated assembly of novel nanoconjugates has been subjected to numerous investigations nowadays, which provides a new insight into the material science and engineering. Via the molecular biology technology, the genetically engineered polypeptide for inorganics (GEPI) can be designed as a molecular binder into the bio-scaffold to assemble hybrid functional nanoarchitectures. In the present work, we constructed a multi-functional virus-like particle (VLP) scaffold based on the Tobacco mosaic virus (TMV) (wild-type strain U1) in a facile way. The S123C site-mutated coat protein (CP) of TMV recombinated with a Ti–GEPI and 6-histidine tag (His tag) was successfully cloned, expressed and purified. The as-produced r-CP products retained both binding affinity for inorganic nanoparticles and self-assembly capability. Analysis of r-CP self-assembly during condensation polymerization was conducted. The r-CP-assembled aggregates including disc-like structures and rod-like VLPs were recovered by preparative size-exclusion chromatography (SEC) and then examined by TEM analysis. Notably, r-CP protein displayed a thermo-triggered reversibly switchable transition between bistable states of transparency and opaqueness. Western blot analysis, Immunogold labelling and TEM analysis were employed to test the existence and binding function of the His tag group in r-CP VLPs. Furthermore, dispersion of TiO2 NPs in r-CP solution and recyclable application in bio-benefication was introduced. Here we demonstrated the possibilities of combining peptide-mediated immobilization with VLP-based biotemplate bearing multi-binding moieties for prospectful functional applications.
    Journal of Materials Science 04/2014; 49(7). · 2.31 Impact Factor
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    ABSTRACT: Enterovirus 71 (EV71) is a highly transmissible pathogenic agent that causes severe central nervous system diseases in infected infants and young children. Here, we reported that EV71 VP1 protein could bind to vimentin intermediate filaments expressed on host cell surface. Soluble vimentin or an antibody against vimentin could inhibit the binding of EV71 to host cells. Accompanied with the reduction of vimentin expression on cell surface, the binding of EV71 to cells was remarkably decreased. Further evidence showed that the N terminal of vimentin is responsible for the interaction between EV71 and vimentin. These results indicated that vimentin on host cell surface may serve as an attachment site that mediated the initial binding and subsequently increased the infectivity of EV71. This study delivers important findings on the roles of vimentin filaments in relation to EV71 infection and provides information that not only improved our understanding of EV71 pathogenesis, but also presents us with potentially new strategies for the treatment of diseases caused by EV71 infections.
    Journal of Virology 03/2014; · 5.08 Impact Factor
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    ABSTRACT: Exposure of cells to type I interferon (IFN) induces an antiviral state that prevents viral infection, but viruses can utilize multiple tactics to antagonize the host immune system. Enterovirus 71 (EV71) and Coxsackievirus A16 (CVA16) are two major pathogens that cause hand, foot, and mouth disease (HFMD), which is prevalent among children. We found that both EV71 and CA16 have different reactions to type I IFN pretreatment and induction patterns of type I IFN on Rhabdomyosarcoma (RD) cells. Further, a human-α and β IFN PCR array was employed to analyze the expressions of 84 genes related to the type I IFN pathway. We found significant up-regulation of multiple genes in the presence of type I IFN and differential regulation patterns during EV71 or CA16 infection in RD cells. For instance, EV71 infection repressed the JAK-STAT signaling pathway and interferon-stimulated gene (ISG) expression, whereas CA16 infection normally triggers the JAK-STAT pathway, leading to the expression of ISGs. Taken together, this study provides a comprehensive view of the differential impacts of EV71 and CA16 infection on 84 genes in the IFN pathway, shedding light on the different resistances of these viruses to type I IFN treatment and cytotoxic effects in RD cells.
    Biochemical and Biophysical Research Communications 01/2014; · 2.41 Impact Factor
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    ABSTRACT: Nuclear proteins can be triggered to be redistributed to the cytoplasm to assist with EV71 virus replication. This process is frequently involved in cellular signal transduction upon virus infection. In this study, we have demonstrated that a new nuclear protein, 68-kDa Src-associated in mitosis protein (Sam68), was translocated to the cytoplasm and was co-localized with EV71 during virus infection. Confocal microscopy and subcellular fractionation assay confirmed that virus 3C protease triggered the redistribution of Sam68 to the cytoplasm. Knockdown of Sam68 expression using ShRNA significantly inhibited virus replication, suggesting that Sam68 may be a host factor involved in EV71 life cycle. In addition, EV71-induced Akt phosphorylation involved a PI3K-dependent mechanism. Sam68 is known to be an upstream regulator of PI3K and our immunoprecipitation studies confirmed that Sam68 interacted directly with the p85 regulatory subunit of PI3K and mediated PI3K/Akt activation during EV71 infection. On the contrary, silencing of Sam68 dramatically abrogated Akt phosphorylation. These data, plus the fact that Sam68 is known to be a signaling adaptor protein, indicated that Sam68 is a signal molecule with a functional role in the PI3K/Akt signal pathway during EV71 infection.
    Virus Research 12/2013; · 2.75 Impact Factor
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    ABSTRACT: Herein, we report the development of efficient inhibitors of reverse transcriptase (RT) of HIV-1 based on indole-alkyl trifluoropyruvate derivatives by a TZM-bl cell assay. The inhibitory activities of the two enantiomers and the corresponding racemic mixture have been compared. TZM-bl cells exhibited strong enantioselective discrimination for the (R)-configuration, among these indole derivatives, the most active compound R-, with a 5-NO2 substituent, gave the best result when tested in the TZM-bl cells on HIV virus type HIV-1IIIB, with an EC50 value of 0.019 μM, CC50 value of 210.697 μM and SI (selectivity index, CC50/EC50) value of 11 089, respectively. The cell test showed that, in most cases, the R-enantiomer was superior to the Rac-mixture, which was better than the corresponding S-enantiomer. The results indicated that the R-enantiomer is the most favorable configuration as an efficient HIV-1 inhibitor. Molecular modeling studies suggested a structural basis for the enantioselectivity of RT towards this class of molecules.
    Organic & Biomolecular Chemistry 11/2013; · 3.57 Impact Factor
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    ABSTRACT: Chronic hepatitis B virus infection is a major cause of hepatic fibrosis, leading to liver cirrhosis and hepatocellular carcinoma. Hepatitis B virus e antigen (HBeAg) is an accessory protein of HBV, not required for viral replication but important for natural infection in vivo. Hepatic stellate cells (HSCs) are the major producers of excessive extracellular matrix during liver fibrogenesis. Therefore, we examined the influence of HBeAg on HSCs. The rat HSC line HSC-T6 was transfected with HBeAg plasmids, and expression of α-smooth muscle actin, collagen I, transforming growth factor-β1 (TGF-β), and tissue inhibitors of metalloproteinase 1 (TIMP-1) was investigated by quantitative real-time PCR. The proliferation of HSCs was determined by MTS analysis. HBeAg transduction induced up-regulation of these fibrogenic genes and proliferation of HSCs. We found that HBeAg induced TGF-β secretion in HSCs, and the activation of HSCs was prevented by a neutralizing anti-TGF-β antibody. Depletion and addition of HBeAg protein in conditioned medium from HSC-T6 cells transduced with HBeAg indicated that HBeAg directly induced the activation and proliferation of rat primary HSCs. Taken together, HBeAg induces the activation and proliferation of HSCs, mainly mediated by TGF-β, and HBeAg protein purified from cell medium can directly activate HSCs.
    Biochemical and Biophysical Research Communications 05/2013; · 2.41 Impact Factor
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    ABSTRACT: A series of chiral gossypol derivatives and its analogs were synthesized and tested in vitro for their anti-H5N1 activity. Interestingly, (+)-gossypol derivatives and its analogs were more active against H5N1 than the corresponding (-)-gossypol derivatives and its analogs. Through a simple chemical modification with amino acids, less active chiral gossypol could be converted into more active derivatives, and most of chiral gossypol derivatives were more potent against H5N1 than 1-adamantylamine. With regard to the mechanism of action, chiral gossypol derivatives and its analogs might impair the virus entry step of cell infection, likely targeting to HA2 protein.
    Bioorganic & medicinal chemistry letters 03/2013; · 2.65 Impact Factor
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    ABSTRACT: Abstract HR212, a recombinant protein composed of the heptad repeat, is a rationally designed human immunodeficiency virus type 1 (HIV-1) fusion inhibitor. This protein can be easily produced by Escherichia coli at a low cost. Previously, studies indicated that HR212 can efficiently inhibit the entry and replication of both laboratory and clinical HIV-1 strains, and this protein is more stable and less sensitive to proteinases than T20. The procedure of HIV-1 entry into the host cells can be divided into three main steps: gp120-CD4 interactions, coreceptor binding, and gp41 six-helix bundle formation and subsequent membrane fusion. The present study demonstrates that HR212 does not block gp120-CD4 binding or interfere with binding to the coreceptors CXCR4 and CCR5. Instead, HR212 efficiently blocks the six-helix bundle formation between peptides derived from the N-terminal heptad repeat (NHR) and the C-terminal heptad repeat (CHR) region of gp41. Fluorescence native polyacrylamide gel electrophoresis (FN-PAGE) indicated that HR212 could form a complex with peptide N36 to block gp41 fusogenic core formation. These results suggest that HR212 inhibits HIV-1 entry by targeting the NHR region of gp41. Therefore, HR212 can potentially be developed as a novel, high-efficiency, specific HIV-1 entry inhibitor.
    AIDS research and human retroviruses 01/2013; · 2.18 Impact Factor
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    ABSTRACT: Amino acids at position 267–298 in E2 protein of GB virus C (GBV-C) were recognized as the antigenic site, and peptides within the region were previously reported to have inhibitory effect on HIV entry. The effect of sequence variability between different types of GBV-C on the antigenic region of the E2 protein was studied by using phylogenetic analysis. Eighty-one unique sequences encompassing this region derived from all seven GBV-C genotypes were compared to each other in this study. The results showed that GBV-C E2 antigenic nucleotide sites belonging to genotype 3 clustered together regardless of synonymous or nonsynonmous sites in the region, whereas, GBV-C E2 antigenic nucleotide sites belonging to the other 6 genotypes clustered together regardless of genotypes. Despite the fact that GBV-C genotypes might confer different degree of ‘protection’ against HIV, the lack of clustering as a unique group based on the amino acid differences in the antigenic site among the six genotypes suggested some other genomic regions or secondary structure of E2 protein might have played a crucial role in determining the variable protection effect of GBV-C on HIV infection.
    Virus Research 01/2013; 178(2):502–505. · 2.75 Impact Factor
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    ABSTRACT: Enterovirus 71 (EV 71) and Coxsackievirus A16 (CA 16) are two major causative agents of hand, foot and mouth disease (HFMD). They have been associated with severe neurological and cardiological complications worldwide, and have caused significant mortalities during large-scale outbreaks in China. Currently, there are no effective treatments against EV 71 and CA 16 infections. We now describe the development of a novel minicircle vector based RNA interference (RNAi) system as a therapeutic approach to inhibiting EV 71 and CA 16 replication. Small interfering RNA (siRNA) molecules targeting the conserved regions of the 3C(pro) and 3D(pol) function gene of the EV 71 and CA 16 China strains were designed based on their nucleotide sequences available in GenBank. This RNAi system was found to effectively block the replication and gene expression of these viruses in rhabdomyosarcoma (RD) cells and virus-infected mice model. The inhibitory effects were confirmed by a corresponding decrease in viral RNA, viral protein, and progeny virus production. In addition, no significant adverse off-target silencing or cytotoxic effects were observed. These results demonstrated the potential and feasibility of this novel minicircle vector based RNAi system for antiviral therapy against EV 71 and CA 16 infection.
    Antiviral research 08/2012; 96(2):234-244. · 3.61 Impact Factor
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    ABSTRACT: Potassium channel tetramerization domain containing 1 (KCTD1) contains a BTB domain, which can facilitate protein-protein interactions that may be involved in the regulation of signaling pathways. Here we describe an expression and purification system that can provide a significant amount of recombinant KCTD1 from Escherichia coli. The cDNA encoding human KCTD1 was amplified and cloned into the expression vector pET-30a(+). The recombinant protein was expressed in E. coli BL21(DE3) cells and subsequently purified using affinity chromatography. To confirm that KCTD1 was correctly expressed and folded, the molecular weight and conformation were analyzed using mass spectroscopy, Western blot, and circular dichroism. Optimizing KCTD1 expression and investigating its secondary structure will provide valuable information for future structural and functional studies of KCTD1 and KCTD family proteins.
    Biochemistry (Moscow) 08/2012; 77(8):941-5. · 1.15 Impact Factor
  • Yeping Sun, Po Tien
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    ABSTRACT: Dynamin, a large guanosine triphosphatase (GTPase), has been implicated in virus entry, but its mechanisms of action are controversial. The entry procedure of most enveloped viruses involves endocytosis and membrane fusion. Dynamin has been suggested to act both as a regulatory GTPase by controlling the early stages of clathrin-mediated endocytosis (CME), which is an important endocytic pathway utilized by many viruses, and as a mechanchemical enzyme that induces membrane fission and pinches endocytic vesicles off from the cellular plasma membrane in later stages in several endocytic pathways, including CME. In addition to its involvement in virus endocytosis, dynamin has also been proposed to participate in membrane fusion between the virus and endosomes following endocytosis. Crystal structures and cryo-electron micrography (cryo-EM) have elucidated the structure of dynamin, which led to development of a mechanochemical model of how dynamin-mediated membrane fission occurs. Based on this, we propose a hypothetical model that explains how dynamin facilitates virus membrane fusion and discuss its roles in virus entry.
    Critical Reviews in Microbiology 06/2012; · 5.07 Impact Factor

Publication Stats

2k Citations
585.30 Total Impact Points

Institutions

  • 2003–2014
    • Wuhan University
      • • State Key Laboratory of Virology
      • • College of Life Sciences
      Wu-han-shih, Hubei, China
  • 1995–2014
    • Chinese Academy of Sciences
      • • Center for Molecular Virology
      • • Institute of Microbiology
      • • Department of Molecular Virology
      Peping, Beijing, China
  • 2013
    • Heilongjiang Bayi Agricultural University
      Sa-erh-t’u, Heilongjiang Sheng, China
  • 1993–2013
    • Northeast Institute of Geography and Agroecology
      • Institute of Microbiology
      Beijing, Beijing Shi, China
    • Beijing Institute of Microbiology and Epidemiology
      Peping, Beijing, China
    • Heinrich-Heine-Universität Düsseldorf
      • Institute of Physical Biology
      Düsseldorf, North Rhine-Westphalia, Germany
  • 2012
    • Renmin University of China
      Peping, Beijing, China
  • 2009–2011
    • Kunming Institute of Zoology CAS
      • State Key Laboratory of Genetic Resources and Evolution
      Yün-nan, Yunnan, China
  • 2008
    • Academia Sinica
      • Institute of Plant and Microbial Biology
      T’ai-pei, Taipei, Taiwan
  • 2007
    • Peking University
      • School of Life Sciences
      Beijing, Beijing Shi, China
  • 2006–2007
    • Case Western Reserve University
      • Department of Pathology (University Hospitals Case Medical Center)
      Cleveland, OH, United States
  • 2004
    • Zhejiang University
      • College of Life Sciences
      Hangzhou, Zhejiang Sheng, China
  • 2003–2004
    • Tsinghua University
      • Laboratory of Structural Biology
      Beijing, Beijing Shi, China
  • 2000
    • Fujian Agricultural University
      China