Enterovirus 71 (EV71) is one of the primary causative agents of hand, foot and mouth disease in children and closely associated with severe neurological complications and even deaths.EV71 outbreaks have occurred throughout the Asia-Pacific region since 1990s, posing global public health threat;however, no specific therapeutic strategy exists for severe EV71 infection.Several inactivated vaccine products have entered or finished the clinical trial stage, and some novel vaccine candidates, including live attenuated, subunit, and virus-like particle, show great potential for further develop-ment.This review summarizes the present situation and progress in the development of EV71 vaccines.

Grass carp reovirus (GCRV), a double stranded RNA virus that infects aquatic animals, often with disastrous effects, belongs to the genus Aquareovirus and family Reoviridea. Similar to other reoviruses, genome replication of GCRV in infected cells occurs in cytoplasmic inclusion bodies, also called viral factories. Sequences analysis revealed the nonstructural protein NS80, encoded by GCRV segment 4, has a high similarity with uNS in MRV(Mammalian orthoreoviruses), which may be associated with viral factory formation. To understand the function of the uNS80 protein in virus replication, the initial expression and identification of the immunogenicity of the GCRV NS80 protein inclusion forming-related region (335.742) was investigated in this study. It is shown that the over-expressed fusion protein was produced by inducing with IPTG at 28oC. In addition, serum specific rabbit antibody was obtained by using super purified recombinant NS80(335.742) protein as antigen. Moreover, the expressed protein was able to bind to anti-his-tag monoclonal antibody (mouse) and NS80(335-742) specific rabbit antibody. Further western blot analysis indicates that the antiserum could detect NS80 or NS80C protein expression in GCRV infected cells. This data provides a foundation for further investigation of the role of NS80 in viral inclusion formation and virion assembly.

Sequences analysis revealed Grass carp reovirus (GCRV) s10 was 909 nucleotides coding a 34 kDa protein denoted as VP7, which was determined to be a viral outer capsid protein (OCP). To obtain expressed OCP in vitro, a full length VP7 gene was produced by RT-PCR amplification, and the amplified fragment was cloned into T7 promoted prokaryotic expression vector pRSET. The recombinant plasmid,which was named as pR/GCRV-VP7,was then transformed into E.coli BL21 host cells. The data indicated that the expressed recombinant was in frame with the N-terminal fusion peptide. The over-expressed fusion protein was produced by inducing with IPTG, and its molecular weight was about 37kDa, which was consistent with its predicted size. In addition, the fusion protein was produced in the form of the inclusion body with their yield remaining steady at more than 60% of total bacterial protein. Moreover,the expressed protein was able to bind immunologically to anti-his-tag monoclonal antibody (mouse) and anti-GCRV serum (rabbit). This work provides a research basis for further structure and function studies of GCRV during entry into cells.

Vaccine immunization represents the most effective strategy against viral infections .Antiviral vaccine candi-dates currently include live attenuated vaccine , inactivated vaccine , DNA vaccine and genetic engineering vaccine .Live at-tenuated vaccines , eliciting efficient humoral and cellular immune response by simulating natural virus infection , are clini-cally widely used.Traditionally, live attenuated vaccine strains are obtained by serial passaging in vitro or in vivo, a costly procedure accumulating random mutations .microRNA( miRNA)-targeting host gene regulation exhibits high species and tis-sue specificity .Introducing certain miRNA target sequence into viral genome by reverse genetic technique can effectively attenuate virus strains, which has great potential in vaccine design and development .In this article,attenuation strategies and progress in its application in vaccine research are disscussed .

ObjectiveTo explore the relationship of marriage quality with individual characteristics and copy style among the epilepsy patients.MethodsThe Chinese Marriage Quality Questionnaire,and Eysenck Personality Questionnaire Short Form Scale-Chinese edition (EPQ-RSC),and the Questionnaire of Simple Way to Coping were assessed used to assess 97 cases of the aduh married epilepsy patients and 93 cases of the control group 1 (epilepsy patients spouses) and 91 cases of the control group 2 (healthy crowds ).ResultsThe total scores of the marriage quality of epilepsy patients ( 295.78 ± 40.37 ) was lower than ones in the control group 2 ( ( 354.85 ± 28.11 ),P =0.000) and higher than that of the control group 1 ( (278.55 ± 42.23 ),P =0.014),the difference was all statistically significant(P＜ 0.05).The total scores of marriage quality of the epilepsy patients was negative correlated with the sex,and the age,and the marriage age and the severity,and the duration,and psychoticism( the Pearson correlation was respectively:-0.204,-0.436,-0.243,-0.691,-0.568,-0.635 ),and was positive correlated with which the formal schooling,and the attack types (symptoms epilepsy),and the introversion or extroversion,and the hidness,and the positive( the Pearson correlation was respectively 0.317,0.750,0.267,0.386,0.188) the difference was all statistically significant (P ＜ 0.05).Those seven variables that the attack type,and the hidness,and the severity,and the sex,and the education,and the psychoticism,and the positive could be predict to marriage quality of epilepsy patients,Adjusted R square was 0.903,After adjustment was 0.021 (P ＜ 0.05 ).ConclusionThe lever of marriage quality of the epilepsy patients is lower,and correlate with severity of epilepsy disease,and personality,copy style,and can be do comprehensive intervention from those.

Objective To prepare quality control samples for St.Louis encephalitis virus(SLEV)molecular detection by constructing pseudovirus containing target sequences of SLEV.Methods According to the principles of armored RNA technique, the prM gene sequence of SLEV was cloned into the prokaryotic expression vector to generate recombinant plasmid pSE380-MS2-SLEV.Then, recombinant E.coli transformed with the corresponding plasmid was induced with IPTG to produce recombinant pseudovirus particles.The particles were purified by chloroform and further characterized by double enzyme digestion and transmission electron microscopy.The temperature sensitivity experiments and quantitative RT-PCR were performed to validate the potential of these pseudovirus particles as quality control samples.Results PCR amplification and sequencing analysis confirmed that the prM gene sequence of SLEV was cloned into vector pSE380-MS2.Transmission electron microscopy showed that homogenous spherical particles with a diameter of about 25 nm were produced upon IPTG induction.The SLEV genomic RNA within the pseudovirus particles was resistant to DNaseⅠand RNase A digestion, and remained stable for 20 days at 37℃.These samples were validated with quantitative RT-PCR for SLEV.Conclusion The RNase-resistant and stable pseudovirus particles containing prM fragment of SLEV are constructed successfully, which can be used as positive quality control samples for RNA extraction and molecular detection.

Influenza, caused by influenza virus, is a serious respiratory illness which poses a global public health threat. Vaccination is the primary strategy for the prevention and control of influenza. Although both inactivated vaccines and the live attenuated vaccines are effective in preventing influenza, the current vaccines have poor efficacy in the elderly and fail to provide protection against heterosubtype viruses. Development of a safer and more effective influenza vaccine that provides broad cross protection, overcoming the intrinsic limitation of the current vaccines, has been a scientific challenge. During the past decades, structural biology, reverse genetic and other virological technologies developed quickly and sped the progress of influenza vaccinology. Some new strategies for developing influenza vaccine have been generated, produced encouraging results, which showed great prospect as next-generation of influenza vaccines.
Disease Outbreaks ; prevention & control ; Hemagglutinin Glycoproteins, Influenza Virus ; immunology ; Humans ; Influenza Vaccines ; biosynthesis ; immunology ; Influenza, Human ; immunology ; prevention & control ; virology ; Orthomyxoviridae ; immunology ; Vaccines, Attenuated ; immunology ; Vaccines, Inactivated ; immunology

Hand-foot-mouth disease (HFMD) is an acute infectious disease caused by various enteroviruses. Recently, large HFMD outbreaks caused by enterovirus type 71 (EV71) have been frequently reported in China, posing great threats on children's health. There is no specific antiviral therapy for severe HFMD, and patient management mainly depends on supportive and symptomatic treatment. Intravenous immunoglobulin (IVIG) is a pharmaceutical preparation of human IgG that is pooled from thousands of healthy blood donors, and contained neutralization antibodies against various enteroviruses, including EV71. IVIG therapy should be carefully administrated for severe HFMD considering its role on passive immunization against EV71 and immune regulation.
Antibodies, Neutralizing ; immunology ; therapeutic use ; Antibodies, Viral ; immunology ; therapeutic use ; Enterovirus A, Human ; immunology ; Hand, Foot and Mouth Disease ; therapy ; virology ; Humans ; Immunization, Passive ; Immunoglobulins, Intravenous ; immunology ; therapeutic use

Objective: To assess the quality for meta-analysis on prevention and treatment of coronary artery disease (CAD) in China.
Methods: We systemically searched 4 Chinese databases of VIP, CNKI, CBM and Wan Fang for their meta-analysis on CAD prevention and treatment from 1987-01 to 2013-10. According to inclusion and exclusion criteria, 2 researchers independently screened and cross-checked all the literatures. The qualities of methodology and report were evaluated by R-AMSTAR and PRISMA scales.
Results: A total of 201 literatures were enrolled for our study. The average score of methodology quality was (24.65±3.97), no literature met all required items, and the major problems were as lack of“a priori design”, insufifcient and bias of data selection combining inappropriate data synthesis. The average score of report quality was (17.20 ± 2.90), no literature met all 27 required items, and the major problems were as incomplete report of abstract, objective, protocol and registration, incomplete data collection/analysis, using and publishing bias information, incomplete quality assessment.
Conclusion: Both of methodology and report of meta-analysis for CAD prevention and treatment have quality problems at different levels, further improvement should be expected.

Viral replicon is a kind of self-replicating viral RNA sourced from viral genome, which contains viral non-structural genes that are critical for viral genome replication with structural proteins deleted or replaced by foreign genes. Kunjin virus is a member of the Flavivirida family, Flavivirus genus, and Kunjin virus replicon is the first and the clearly defined flavivirus replicon. Kunjun virus replicon has been regarded as an excellent viral vector on account of its high expression, lower cytotoxicity and genetic stability. These unique characteristics of kunjin virus replicons make them suitable for the study of viral genome replication, recombinant proteins production, vaccine development and gene therapy. In this article, recent progress in the development, properties and applications of kunjin virus replicon system was briefly reviewed.
Genetic Vectors ; genetics ; Genome, Viral ; Recombination, Genetic ; Replicon ; genetics ; Virus Replication ; physiology ; West Nile virus ; genetics ; metabolism