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Interferon-induced Transmembrane Proteins (IFITMs) were identified through small interference RNA (siRNA) screening method in 1980s. The antiviral properties of the IFITMs were firstly discovered in 1996. Recently, its antiviral effect and mechanism have become a research hotspot. Many studies have shown that IFITM can inhibit the replication of multiple pathogenic viruses, including influenza A virus (IAV), Human Immunodeficiency Virus (HIV-1), hepatitis C virus (HCV), Ebola virus (EBOV), West Nile virus and so on. IFITMs inhibit the replication of virus in the early stage of the viral life cycle, which occurred before the release of viral genomes into the cytosol. Recent studies indicate that IFITM proteins could block viral replication by mediate viral membrane fusion. However, the mechanism is still under investigation. Here we review the discovery and characterization of the IFITM proteins, elucidate their antiviral activities and the potential mechanisms.
Animals ; Humans ; Interferons ; genetics ; immunology ; Membrane Proteins ; genetics ; immunology ; Virus Diseases ; genetics ; immunology ; virology ; Viruses ; genetics ; immunology

Objective To explore origin recognition complex 1(ORC1) expression in the rat vascular muscle cells at different phases of proliferation.Methods Vascular smooth muscle cells(VSMCs) of thoracic aorta of rats in primary culture were obtained by the adherence method of tissue culture.Total RNA of VSMCs was extracted.The expression of ORC1 mRNA of VSMCs at different phases of proliferation was determined by reverse transcription polymerase chain reaction(RT-PCR) and the expression of ORC1 protein by immunocytochemistry and laser confocal microscopy.Results Cultured VSMCs were confirmed by light microscope and immunocytochemistry.The expression of ORC1 mRNA in the quiescence stage of VSMCs was not found.After VSMCs were stimulated with serum,the level of ORC1mRNA had an obvious increase at 6 h,peaked at 12 to 24 h and decreased in the following 24 h.The expression of ORC1 protein was also not found in the quiescence stage of VSMCs,but the level of ORC1 protein during proliferation of VSMCs was significantly increased.Conclusion ORC1 may have an important role during the process of VSMCs proliferation in rats.

Objective To explore the expression of origin recognition complex1(ORC1) during DNA replication progress of rat vascular muscle cells(VSMCs).Methods VSMCs of rat thoracic aorta were obtained by the adherence method of tissue culture.The growth curve was drawn by MTT.The association between DNA replication and the expression of ORC1 mRNA and protein in different growth phases of VSMCs was analyzed.Results The expression of ORC1 mRNA and protein in quiescence stage of VSMCs was not found.After stimulated with serum,the expression of ORC1 mRNA in rat VSMCs increased significantly,peaked at 12-24 h.The expression of ORC1 protein was similar to ORC1 mRNA in VSMCs.Meanwhile,the higher DNA replication of stimulated VSMCs was observed,peaked at 12-24 h after serum addition. Conclusion ORC1 may be involved in the DNA replication of rat VSMCs during the progress of proliferation.

Objective To investigate the influence of RNA interference targeting ORC1 gene on the proliferation of rat vascular smooth muscle cells(VSMCss).Methods VSMCss were transfected with siRNA targeting ORC1 gene by liposome.The expression of ORC1 protein was detected by Western blotting.MTT test and ~(3)H thymidine~()(~(3)H-TdR) incorporation were used to detect VSMCss proliferation.The expression of proliferating cell nuclear antigen(PCNA) in VSMCss was detected by immunocytochemistry.Results After transfected of the three pairs of siRNA targeting ORC1 genes respectively,the expression of ORC1 was all lower than that in the control group(non-transfection and negative siRNA).Especially,that in the group transfected of the second pair positive siRNA decreased most significantly.The optical density of MTT,~(3)H thymidine incorporation and the expression of PCNA decreased significantly in VSMCss transfected with siRNA targeting ORC1 gene as compared with that in the control group.Conclusion ORC1 gene silenced by RNA interference can inhibit VSMCss proliferation.

The microencapsulated genetic cells may be a new platform instead of genetic engineering drugs, as they can overcome the genetic engineering drugs' shortages such as short half-life in vivo, low activity, and incomplete elimination of organic solvent. This article reviews and summarizes the advantages, possible problems and solution and the feasibility of using microencapsulated genetic engineering cells in the treatment of cancer.
Animals ; Capsules ; Cell Transplantation ; Combined Modality Therapy ; Drug Compounding ; methods ; Genetic Engineering ; Genetic Therapy ; methods ; Mice ; Neoplasms, Experimental ; therapy ; Transfection

Preparation of maternal strain A/PR/8/34 HA antiserum for influenza virus classical reassortment. A/PR/8/34 virus was digested by bromelain after inactivation and purification. 5%-20% sucrose continuous density gradient centrifugation method was used to purify HA protein. SIRD method was used to select the target protein. SDS-PAGE method was used to identified HA protein. High Immunogenic A/PR/8/34 HA protein was successfully prepared and HI titer reached 10240. High purity HA antiserum was identified by SIRD method. The key reagent in the classical reassortment of influenza virus was prepared, and the complete set of technical methods were explored, which laid the foundation for the independent research and development of seasonal influenza vaccine strains of China.
Animals ; Antibodies, Viral ; immunology ; Electrophoresis, Polyacrylamide Gel ; Female ; Hemagglutination Inhibition Tests ; Hemagglutinin Glycoproteins, Influenza Virus ; analysis ; immunology ; Humans ; Influenza A Virus, H1N1 Subtype ; genetics ; immunology ; Influenza, Human ; immunology ; virology ; Rabbits ; Reassortant Viruses ; genetics ; immunology

Abstract: To investigate the distribution of avian influenza virus in environmental samples from live poultry markets (LPM) in China, samples were collected and tested by nucleic acid during 2009-2013 season. Each sample was tested by real-time RT PCR using flu A specific primers. If any real-time PCR was positive, the sample was inoculated into specific-pathogen-free (SPF) embryonated chicken eggs for viral isolation. The results indicated that the positive rate of nucleic acid in enviromental samples exhibited seasonality. The positive rate of nucleic acid was significantly higher in Winter and Spring. The positive rate of nucleic acid in LPM located in the south of China was higher than in northern China. Samples of Sewage for cleaning poultry and chopping board showed that higher positive rate of nucleic acid than other samples. The Subtype identification showed that H5 and H9 were main subtypes in the enviromental samples. Viral isolation indicated H5 subtypes was more than H9 subtypes between 2009 and 2013 while H9 subtypes increased in 2013. Our findings suggested the significance of public health based on LPM surveillance and provided the basis of prevention and early warning for avian flu infection human.
Animals ; China ; Feces ; virology ; Fresh Water ; virology ; Influenza A virus ; classification ; genetics ; isolation & purification ; Influenza in Birds ; virology ; Poultry ; Poultry Diseases ; virology ; Public Health ; Seasons ; Sewage ; virology

Hemagglutinin (HA) contains a head domain with a high degree of variability and a relatively conserved stem region. HA is the major viral antigen on the surface of the influenza virus. To define the biologic activities of chimeric HA bearing different head domains and stem regions or their potential use, a HA chimeric gene containing the head domain of the H7 subtype virus and stem region of the H3 subtype virus was modified and expressed using a baculovirus expression vector. Then, the secreted protein was purified and its biologic activities characterized. Approximately 1.4 mg/mL cH7/3 HA could be obtained, and its molecular weight was ≈ 70 kD. The trimer form of cH7/3 protein had hemagglutination activity and could be recognized by specific antibodies. The method described here can be used for further studies on the screening of HA stem-reactive antibodies or the development of vaccines with conserved epitopes.
Antibodies, Viral ; immunology ; Baculoviridae ; genetics ; metabolism ; Gene Expression ; Genetic Vectors ; genetics ; metabolism ; Hemagglutination ; Hemagglutinin Glycoproteins, Influenza Virus ; genetics ; immunology ; Humans ; Influenza Vaccines ; genetics ; immunology ; Influenza, Human ; prevention & control ; virology

Objective To investigate the possibility of using well-differentiated human airway epithelial cells (HAE) to isolate and identify human influenza A virus from a stale respiratory tract specimen.Methods The stale specimen used in this study was a nasopharyngeal swab specimen collected from a patient with unexplained pneumonia in Qinghai in 2010.It was positive for influenza A virus (H3N2) RNA, but negative for hemagglutination.Equal amount of the specimen was inoculated on HAE and on Madin-Darby canine kidney (MDCK) cells for virus isolation and passage.Cytopathic effects were observed daily after inoculation.Hemagglutination inhibition test was performed at every passage.Electron microscope was used to observe viral morphology.Viral genome was sequenced, followed by molecular evolutionary analysis.Results No progeny virus was isolated in MDCK cells, while a influenza A virus subtype H3N2 strain [A/Qinghai/178/2010(H3N2)] was isolated in HAE with a typical morphology and cytopathic effect of influenza A infection.The hemagglutination inhibition activity was 1∶16.Results of the molecular evolutionary analysis of viral genome showed that the influenza A virus (H3N2) strain was highly homologous to the A/Nanjing/1655/2010(H3N2) strain, which was isolated during the 2010 influenza pandemic in Nanjing.Conclusion HAE can be used for isolation and identification of virus from stale respiratory tract specimens.It is more sensitive than MDCK cells with regard to human influenza virus isolation.