Japanese Encephalitis Virus (JEV) is responsible for over 30,000 annual cases of encephalitis worldwide, causing 30% mortality. JEV is thus a continuing threat to public health, so development of new antiviral drugs a...Japanese Encephalitis Virus (JEV) is responsible for over 30,000 annual cases of encephalitis worldwide, causing 30% mortality. JEV is thus a continuing threat to public health, so development of new antiviral drugs against JEV is desirable. Here, we examined JEV replication in mouse and used a short hairpin RNA JRi as the antiviral agent. The features of virus replication in neuron and survival rates of mice infected with JEV were different between virus strains. The mice infected with the virulent JEV strain (JaGAr01) were injected with pJRi (100 μg/mouse) which produced shRNAJRi. The survival rates of mice treated at 3 days before, the same day and 3 days after JEV infection were 22%, 78% and 44%, respectively. In addition, we demonstrated that the injection of pJRi induced interferon (IFN) production in cells and mice. These results suggest that the replication of JEV can be efficiently inhibited by RNAi and innate immunity including IFN. These data mean that pJRi has the inhibitory activity against JEV infection in vivo, and could be used as an antiviral drug to treat JEV infection.展开更多
[ Objective] This study aimed to establish a rapid, sensitive and specific method using reverse transcription loop-mediated isothermal amplification （RT-LAMP） technology to detect swine Japanese B encephalitis virus...[ Objective] This study aimed to establish a rapid, sensitive and specific method using reverse transcription loop-mediated isothermal amplification （RT-LAMP） technology to detect swine Japanese B encephalitis virus （JEV）. [ Method ] Four specific LAMP primers were designed according to six loci the conservative region of JEV E gene sequence. Positive JEV RNA sample was used as a template for one-step amplification, and the reaction conditions and reaction system were optimized. [ Result] Experimental results showed that the established method had high sensitivity, with the detection limit of 0.5pg; specificity experi- ments indicated that the method had high specificity and there was no amplification reaction for other viral pathogens. The coincidence rate between detection results of RT-LAMP and RT-Nested-PCR was 90.9%. After RT-LAMP reaction, a chemiluminescent agent was added for visual observation, which greatly reduced the detection time. This method required no special equipment but only a water bath, which was a simple, sensitive and rapid detection method for swine Japanese B encephalitis virus and could be applied in primary laboratories. [ Conclusion] An RT-LAMP detection method for swine Japanese B encephalitis virus was successfully established and preliminarily applied in clinical practice.展开更多
An animal model of epidemic(Japanese)B encephalitis was estabilisged by injecting theJin Wei Yah 1 stain of B encephalitis virus into the peritoneal cavity of mice.The ultrastructuralchanges of the nerve cells in thei...An animal model of epidemic(Japanese)B encephalitis was estabilisged by injecting theJin Wei Yah 1 stain of B encephalitis virus into the peritoneal cavity of mice.The ultrastructuralchanges of the nerve cells in their brains were studied,special attention being paid to some types ofnerve cell in the cerebellar cortex.The infectet Purkinje cells and especially the granular cells showedspecial and inter,sting pathological features.These were compared with the changes found in the in-fected nerve cells in the cerebral cortex,diencephalon and mesencephalon.A radiating structure consisting of a microveside-microtubule aggregation body at the centerand endoplastic reticulum or virus replication multivesicular structures around it was often found in thein fected nerve cells.Its morphological features were described in detail,and its significance and the se-quenoe of events of its development discussed.In the late stage of infection,virus particles were found in the nuclei of part of the necroticcells.It is considered that they entered the nuclei from the cytoplasm during or after the death of theinfected cells.The observation smade in this study have comfimed in the granular cell of the cerebellum theidea of Chert et al.that the encephalitis B virus particle can he formed in the perinudear cistem ofthe infected nerve cell,and have brought forth further information in this respect.The way of releaseof the virus particles from the infected nerve cells observed in this study is fundamentally consistentwith that observed by Chen et al.but most of the virus particles left the nerve cell via the cell pro-展开更多
文摘Japanese Encephalitis Virus (JEV) is responsible for over 30,000 annual cases of encephalitis worldwide, causing 30% mortality. JEV is thus a continuing threat to public health, so development of new antiviral drugs against JEV is desirable. Here, we examined JEV replication in mouse and used a short hairpin RNA JRi as the antiviral agent. The features of virus replication in neuron and survival rates of mice infected with JEV were different between virus strains. The mice infected with the virulent JEV strain (JaGAr01) were injected with pJRi (100 μg/mouse) which produced shRNAJRi. The survival rates of mice treated at 3 days before, the same day and 3 days after JEV infection were 22%, 78% and 44%, respectively. In addition, we demonstrated that the injection of pJRi induced interferon (IFN) production in cells and mice. These results suggest that the replication of JEV can be efficiently inhibited by RNAi and innate immunity including IFN. These data mean that pJRi has the inhibitory activity against JEV infection in vivo, and could be used as an antiviral drug to treat JEV infection.
基金Supported by Key Scientific and Technological Project of Guangxi Province(GKG 1123007-3)Special Fund for Agro-scientific Research in the Public Interest(201103008,2008030152GX)+2 种基金Scientific Research Project of Fishery,Animal Husbandry and Veterinary Bureau of Guangxi Zhuang Autonomous Region(GYM[08]283219)Natural Science Foundation of Guangxi Zhuang Autonomous Region(2011GXNSFB018032)Systematic Research Project of Guangxi Key Laboratory of Animal Vaccines and New Technology(12-071-28-A-5)
文摘[ Objective] This study aimed to establish a rapid, sensitive and specific method using reverse transcription loop-mediated isothermal amplification （RT-LAMP） technology to detect swine Japanese B encephalitis virus （JEV）. [ Method ] Four specific LAMP primers were designed according to six loci the conservative region of JEV E gene sequence. Positive JEV RNA sample was used as a template for one-step amplification, and the reaction conditions and reaction system were optimized. [ Result] Experimental results showed that the established method had high sensitivity, with the detection limit of 0.5pg; specificity experi- ments indicated that the method had high specificity and there was no amplification reaction for other viral pathogens. The coincidence rate between detection results of RT-LAMP and RT-Nested-PCR was 90.9%. After RT-LAMP reaction, a chemiluminescent agent was added for visual observation, which greatly reduced the detection time. This method required no special equipment but only a water bath, which was a simple, sensitive and rapid detection method for swine Japanese B encephalitis virus and could be applied in primary laboratories. [ Conclusion] An RT-LAMP detection method for swine Japanese B encephalitis virus was successfully established and preliminarily applied in clinical practice.
文摘An animal model of epidemic(Japanese)B encephalitis was estabilisged by injecting theJin Wei Yah 1 stain of B encephalitis virus into the peritoneal cavity of mice.The ultrastructuralchanges of the nerve cells in their brains were studied,special attention being paid to some types ofnerve cell in the cerebellar cortex.The infectet Purkinje cells and especially the granular cells showedspecial and inter,sting pathological features.These were compared with the changes found in the in-fected nerve cells in the cerebral cortex,diencephalon and mesencephalon.A radiating structure consisting of a microveside-microtubule aggregation body at the centerand endoplastic reticulum or virus replication multivesicular structures around it was often found in thein fected nerve cells.Its morphological features were described in detail,and its significance and the se-quenoe of events of its development discussed.In the late stage of infection,virus particles were found in the nuclei of part of the necroticcells.It is considered that they entered the nuclei from the cytoplasm during or after the death of theinfected cells.The observation smade in this study have comfimed in the granular cell of the cerebellum theidea of Chert et al.that the encephalitis B virus particle can he formed in the perinudear cistem ofthe infected nerve cell,and have brought forth further information in this respect.The way of releaseof the virus particles from the infected nerve cells observed in this study is fundamentally consistentwith that observed by Chen et al.but most of the virus particles left the nerve cell via the cell pro-
