Hand foot and mouth disease is a febrile sickness complex characterized by cutaneous eruption (exanthem) on the palms and soles with simultaneous occurrence of muco-cutanous vesiculo-ulcerative lesions (enanthem) affe...Hand foot and mouth disease is a febrile sickness complex characterized by cutaneous eruption (exanthem) on the palms and soles with simultaneous occurrence of muco-cutanous vesiculo-ulcerative lesions (enanthem) affecting the mouth. The illness is caused by a number of enteroviruses with coxsackievirus A16 and enterovirus 71 as the main causative agents. Human enterovirus 71 (EV71) belongs to the species Human enterovirus A under the genus Enterovirus within the family Picornaviridae. EV71 has been associated with an array of clinical diseases including hand foot and mouth disease (HFMD), aseptic meningitis, encephalitis and poliomyelitis-like acute flaccid paralysis. A large outbreak of HFMD due to highly neurovirulent EV71 emerged in Malaysia in 1997, and caused 41 deaths amongst young children. In late 2000, a recurrence of an outbreak of HFMD occurred in Malaysia with 8 fatalities in peninsular Malaysia. Outbreak of HFMD due to EV71 recurred in 2003 with an unknown number of cases and mortalities. A similar outbreak of HFMD with 2 recorded deaths in young children occurred in peninsular Malaysia in late 2005 and this was followed by a larger outbreak in Sarawak (Malaysian Borneo) with 6 reported fatalities in the early part of 2006. The current on-going outbreak of HFMD started in peninsular Malaysia in epidemiological week 12 of 2010. As with other HFMD outbreaks in Malaysia, both EV71 and CA16 were the main aetiological viruses isolated. In similarity with the HFMD outbreak in 2005, the isolation of CA16 preceded the appearance of EV71. Based on the VP1 gene nucleotide sequences, 4 sub-genogroups of EV71 (C1, C2, B3 and B4) co-circulated and caused the outbreak of hand, foot and mouth disease in peninsular Malaysia in 1997. Two sub-genogroups (C1 and B4) were noted to cause the outbreak in 2000 in both peninsular Malaysia and Sarawak. EV71 of sub-genogroup B5 with smaller contribution from sub-genogroup C1 caused the outbreak in 2003. In the 2005 outbreak, besides the EV71 strains of sub-genogroup C1, EV71展开更多
AIM： Enterovirus 71 （EV71） has been implicated as the etiological agent responsible for the recent outbreaks of hand, foot and mouth disease associated with severe neurological diseases in the Asia-Pacific region. ...AIM： Enterovirus 71 （EV71） has been implicated as the etiological agent responsible for the recent outbreaks of hand, foot and mouth disease associated with severe neurological diseases in the Asia-Pacific region. METHODS： The assembly process was hypothesized to occur via an orchestrated proteolytic processing of the P1 precursor by the viral protease 3CD. To test this hypothesis, we constructed 3 recombinant baculoviruses： Bac-P1 expressing P1; Bac-3CD expressing 3CD; and Bac-P1-3CD co-expressing P1 and 3CD. RESULTS： Both single infection by Bac-P1-3CD and coinfection by Bac-P1 and Bac-3CD resulted in correct cleavage of P1 to yield individual proteins VP0, VP1 and VP3, while the former approach yielded higher VLP production. In the cells, the structural proteins selfassembled into clusters of virus-like particles （VLP） resembling the authentic EV71 particle aggregates. After ultracentrifugation purification, the dispersed VLPs were indistinguishable from the authentic virus in size, appearance, composition and surface epitopes, as determined by SDS-PAGE, Western blot, transmission electron microscopy and immunogold labeling. CONCLUSION： Our data, for the first time, suggest that in insect cells EV71 structural proteins adopt a processing and assembly pathway similar to poliovirus assembly. The preservation of particle morphology and composition suggest that the VLP may be a valuable vaccine candidate to prevent EV71 epidemics.展开更多
Background Enterovirus 71 （EV71） and coxsackievirus A16 （Cox A16） are major causative agents for hand, foot and mouth disease （HFMD）. Studies indicate that the frequent HFMD outbreaks result in a few hundreds ch...Background Enterovirus 71 （EV71） and coxsackievirus A16 （Cox A16） are major causative agents for hand, foot and mouth disease （HFMD）. Studies indicate that the frequent HFMD outbreaks result in a few hundreds children＇s death in China in recent years. The vaccine and other research for HFMD need to be developed urgently. The aims of our study were： to explore dynamic development of mother-source neutralizing antibodies against EV71 and Cox A16 in infants from Jiangsu Province, China, and to provide the fundamental data for further establishing of corresponding immunization course. Methods Peripheral blood samples were collected from 133 of parturient women once immediately before delivery and their infants at two and seven months of age. Method of micro-dose cytopathogenic effect was used to measure neutralizing antibodies against EV71 and Cox A16, respectively. Results Seropositive rates of anti-EV71 and anti-Cox A16 in prenatal women were 79.7% （106/133） and 92.5% （123/133）, respectively; geometric mean titers （GMTs） were 29.0 and 61.9; 75.9% （101/133） prenatal women were both positive in anti-EV71 and anti-Cox A16; seropositive rates of anti-EV71 and anti-Cox A16 were 25.6% （34/133） and 38.3% （51/133） in infants at two months of age; GMTs were 12.3 and 18.0, respectively. GMTs of anti-EV71 were significantly higher for infants at seven months （82.6） compared with that at two months （P 〈0.05）, showing infants had inapparently infected by EV71 during two to seven months. Although only one offspring （0.75%） at seven months was found having anti-Cox A16 transfered from maternal, this observation suggested no maternal antibody may remain in infants at seven months. Conclusions The prevalence of EV71 and Cox A16 were relatively high in Jiangsu Province. Bivalent vaccine against both EV71 and Cox A16 should be developed, and the ideal time point for prime immunization for infants is around 2-5 months of age.展开更多
Enterovirus 71(EV71),one of the major causative agents for hand-foot-and-mouth disease(HFMD),has caused more than 100 deaths among Chinese children since March 2008.The EV71 genome encodes an RNAdependent RNA polymera...Enterovirus 71(EV71),one of the major causative agents for hand-foot-and-mouth disease(HFMD),has caused more than 100 deaths among Chinese children since March 2008.The EV71 genome encodes an RNAdependent RNA polymerase(RdRp),denoted 3D^(pol),which is central for viral genome replication and is a key target for the discovery of specific antiviral therapeutics.Here we report the crystal structures of EV71 RdRp(3D^(pol))and in complex with substrate guanosine-5'-triphosphate and analog 5-bromouridine-5'-triphosphate best to 2.4Åresolution.The structure of EV71 RdRp(3D^(pol))has a wider open thumb domain compared with the most closely related crystal structure of poliovirus RdRp.And the EV71 RdRp(3D^(pol))complex with GTP or Br-UTP bounded shows two distinct movements of the polymerase by substrate or analogue binding.The model of the complex with the template:primer derived by superimposition with foot-and-mouth disease virus(FMDV)3D/RNA complex reveals the likely recognition and binding of template:primer RNA by the polymerase.These results together provide a molecular basis for EV71 RNA replication and reveal a potential target for anti-EV71 drug discovery.展开更多
Enterovirus 71(EV71) is one of the main pathogens that causes hand-foot-and-mouth disease(HFMD). HFMD caused by EV71 infection is mostly self-limited; however, some infections can cause severe neurological diseases, s...Enterovirus 71(EV71) is one of the main pathogens that causes hand-foot-and-mouth disease(HFMD). HFMD caused by EV71 infection is mostly self-limited; however, some infections can cause severe neurological diseases, such as aseptic meningitis, brain stem encephalitis, and even death. There are still no effective clinical drugs used for the prevention and treatment of HFMD. Studying EV71 protein function is essential for elucidating the EV71 replication process and developing anti-EV71 drugs and vaccines. In this review, we summarized the recent progress in the studies of EV71 noncoding regions(50 UTR and 30 UTR) and all structural and nonstructural proteins, especially the key motifs involving in viral infection, replication, and immune regulation. This review will promote our understanding of EV71 virus replication and pathogenesis, and will facilitate the development of novel drugs or vaccines to treat EV71.展开更多
The comparative analysis of the biological characterization and the genetic background study of EV71 circulating strains is commonly recognized as basic work necessary for development of an effective EV71 vaccine. In ...The comparative analysis of the biological characterization and the genetic background study of EV71 circulating strains is commonly recognized as basic work necessary for development of an effective EV71 vaccine. In this study, we sequenced five EV71 circulating strains, isolated from Fuyang, Hefei, Kunming and Shenzhen city of China and named them FY-23, FY-22, H44, K9 and S1 respectively. The sequence alignment demonstrated their genotypes be C4. The genetic distance of the VP1 gene from these isolates suggested that they were highly co-related with genetic identity similar to other previously reported EV71 strains in China. Additionally, these strains were identified to display some obvious proliferation dynamics and plaque morphology when propagated in Vero cells. However, a distinctive difference in pathogenic ability in neonatal mice was found. Some differences in cross neutralization test & immunogenic analysis were also found. All these results are related to the biological characterization of circulating EV71 strains in China and aid in the development of an EV71 vaccine in the future.展开更多
Human enterovirus 71(HEV71) is the cause of hand,foot and mouth disease and associated neurological complications in children under five years of age.There has been an increase in HEV71 epidemic activity throughout th...Human enterovirus 71(HEV71) is the cause of hand,foot and mouth disease and associated neurological complications in children under five years of age.There has been an increase in HEV71 epidemic activity throughout the Asia-Pacific region in the past decade,and it is predicted to replace poliovirus as the extant neurotropic enterovirus of highest global public health significance. To date there is no effective antiviral treatment and no vaccine is available to prevent HEV71 infection. The increase in prevalence, virulence and geographic spread of HEV71 infection over the past decade provides increasing incentive for the development of new therapeutic and prevention strategies against this emerging viral infection. The current review focuses on the potential, advantages and disadvantages of these strategies. Since the explosion of outbreaks leading to large epidemics in China, research in natural therapeutic products has identified several groups of compounds with anti-HEV71 activities. Concurrently, the search for effective synthetic antivirals has produced promising results. Other therapeutic strategies including immunotherapy and the use of oligonucleotides have also been explored. A sound prevention strategy is crucial in order to control the spread of HEV71. To this end the ultimate goal is the rapid development, regulatory approval and widespread implementation of a safe and effective vaccine. The various forms of HEV71 vaccine designs are highlighted in this review. Given the rapid progress of research in this area, eradication of the virus is likely to be achieved.展开更多
A sensitive reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for human enterovirus 71 (EV71) and Coxsackievirus A16 (CVA16) infection was further evaluated. The one step reaction was perfor...A sensitive reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for human enterovirus 71 (EV71) and Coxsackievirus A16 (CVA16) infection was further evaluated. The one step reaction was performed in a single tube at 65?C for 45 min for EV71 and 35 min for CVA16. The detection limits of RT-LAMP assays for both EV71 and CVA16 were 0.1 of a 50% tissue culture infective dose (TCID50) per reaction, based on 10—Fold dilutions of a titrated EV71 or CVA16 strain. The specific assay showed there were no cross-reactions with Coxsackievirus A (CVA) viruses (CVA 2, 4, 5, 7, 9, 10, 14, and 25), Coxsackievirus B (CVB) viruses (CVB 1, 2, 3, 4, and 5) or ECHO viruses (ECHO 3, 6, 11, and 19). In parallel with commercial quantitative real-time polymerase chain reaction (qRT-PCR) diagnostic kits for EV71 and CVA16, the RT-LAMP assay was evaluated with 515 clinical specimens, the results showed the RT-LAMP assay and the qRT-PCR assay were in complete agreement for 513/515 (99.6%) of the specimens. Two samples with discrepant results from two methods were further verified by nested reverse transcription polymerase chain reaction (nRT-PCR) assay and sequencing to be true positives for CVA16. In conclusion, RT-LAMP assay is demonstrated to be a sensitive and specific assay and have a great potential for the rapid and visual screening of EV71 and CVA16 in China, especially in those resource-limited hospitals and rural clinics of provincial and municipal regions.展开更多
Human enterovirus 71 viruses have been long circulating throughout the world. In this study, we performed a positive selection analysis of the VP1 genes of capsid proteins from Enterovirus 71 viruses. Our results show...Human enterovirus 71 viruses have been long circulating throughout the world. In this study, we performed a positive selection analysis of the VP1 genes of capsid proteins from Enterovirus 71 viruses. Our results showed that although most sites were under negative or neutral evolution, four positions of the VP1 genes were under positive selection pressure. This might account for the spread and frequent outbreaks of the viruses and the enhanced neurovirulence. In particular, position 98 might be involved in neutralizing antibodies, modulating the virus-receptor interaction and enhancing the virulence of the viruses. Moreover, both positions 145 and 241 might correlate to determine the receptor specificity. However, these positions did not display much difference in amino acid polymorphism. In addition, no position in the VP1 genes of viruses isolated from China was under positive selection.展开更多
This article is the tenth series of the Fungal Diversity Notes,where 114 taxa distributed in three phyla,ten classes,30 orders and 53 families are described and illustrated.Taxa described in the present study include ...This article is the tenth series of the Fungal Diversity Notes,where 114 taxa distributed in three phyla,ten classes,30 orders and 53 families are described and illustrated.Taxa described in the present study include one new family(viz.Pseudoberkleasmiaceae in Dothideomycetes),five new genera(Caatingomyces,Cryptoschizotrema,Neoacladium,Paramassaria and Trochilispora)and 71 new species,(viz.Acrogenospora thailandica,Amniculicola aquatica,A.guttulata,Angustimassarina sylvatica,Blackwellomyces lateris,Boubovia gelatinosa,Buellia viridula,Caatingomyces brasiliensis,Calophoma humuli,Camarosporidiella mori,Canalisporium dehongense,Cantharellus brunneopallidus,C.griseotinctus,Castanediella meliponae,Coprinopsis psammophila,Cordyceps succavus,Cortinarius minusculus,C.subscotoides,Diaporthe italiana,D.rumicicola,Diatrypella delonicis,Dictyocheirospora aquadulcis,D.taiwanense,Digitodesmium chiangmaiense,Distoseptispora dehongensis,D.palmarum,Dothiorella styphnolobii,Ellisembia aurea,Falciformispora aquatic,Fomitiporia carpinea,F.lagerstroemiae,Grammothele aurantiaca,G.micropora,Hermatomyces bauhiniae,Jahnula queenslandica,Kamalomyces mangrovei,Lecidella yunnanensis,Micarea squamulosa,Muriphaeosphaeria angustifoliae,Neoacladium indicum,Neodidymelliopsis sambuci,Neosetophoma miscanthi,N.salicis,Nodulosphaeria aquilegiae,N.thalictri,Paramassaria samaneae,Penicillium circulare,P.geumsanense,P.mali-pumilae,P.psychrotrophicum,P.wandoense,Phaeoisaria siamensis,Phaeopoacea asparagicola,Phaeosphaeria penniseti,Plectocarpon galapagoense,Porina sorediata,Pseudoberkleasmium chiangmaiense,Pyrenochaetopsis sinensis,Rhizophydium koreanum,Russula prasina,Sporoschisma chiangraiense,Stigmatomyces chamaemyiae,S.cocksii,S.papei,S.tschirnhausii,S.vikhrevii,Thysanorea uniseptata,Torula breviconidiophora,T.polyseptata,Trochilispora schefflerae and Vaginatispora palmae).Further,twelve new combinations(viz.Cryptoschizotrema cryptotrema,Prolixandromyces australi,P.elongatus,P.falcatus,P.longispinae,P.microveliae,P.neoalardi,P.polhemorum,P.protubera展开更多
Background:Hand-foot-and-mouth disease(HFMD)is a common pediatric infectious disease caused by a variety of intestinal viruses.Enterovirus 71(EV71)is the primary pathogen that might cause severe symptoms and even deat...Background:Hand-foot-and-mouth disease(HFMD)is a common pediatric infectious disease caused by a variety of intestinal viruses.Enterovirus 71(EV71)is the primary pathogen that might cause severe symptoms and even death in children with HFMD.This study aimed to investigate the intestinal detoxification time of HFMD children with EV71 infection and its related factors.Methods:Sixty-five HFMD children with EV71 infection were followed up.Their stool samples were collected once every 4 to 7 days.Viral nucleic acids were detected byfl uorescent polymerase chain reaction until the results became negative.The positive rates of viral nucleic acids were analyzed by the Kaplan-Meier method.The Log-rank test and Cox-Mantel test were used to analyze factors affecting the HFMD children with EV71 infection.Results:On the 2nd,4th,6th and 10th week,the positive rates of viral nucleic acids in stool samples of the 65 children were 94.6%,48.1%,17.2%and 0,respectively.Univariate analysis showed that the intestinal detoxification time of the children were related to gender,pre-admission disease course,severity of disease,and use of steroids or gamma globulin(P<0.05).Multivariate analysis showed that the severity of disease was an independent factor affecting the intestinal detoxification time(P<0.05),with a relative risk of 2.418.Conclusions:The longest intestinal detoxification time of HFMD children with EV71 infection was 10 weeks.The severity of disease was an important factor affecting the intestinal detoxification time of HFMD children with EV71 infection.Severe HFMD children with EV71 infection had a longer intestinal detoxification time.展开更多
Background:During Enterovirus type 71(EV71)infection,the structural viral protein 1(VP1)activates endoplasmic reticulum(ER)stress associated with peripheral myelin protein 22(PMP22)accumulation and induces autophagy.H...Background:During Enterovirus type 71(EV71)infection,the structural viral protein 1(VP1)activates endoplasmic reticulum(ER)stress associated with peripheral myelin protein 22(PMP22)accumulation and induces autophagy.However,the specific mechanism behind this process remains elusive.Methods:In this research,we used the VP1-overexpressing mouse Schwann cells(SCs)models co-transfected with a PMP22 silencing or Autocrine motility factor receptor(AMFR/gp78)overexpressing vector to explore the regulation of gp78 on PMP22 and its relationship with autophagy and apoptosis.Results:The activity of gp78 could be influenced by EV71-VP1,leading to a decrease in the ubiquitination and degradation of PMP22,resulting in PMP22 accumulation in ER.In VP1-overexpressing mouse SCs,all three ER stress sensors,including pancreatic endoplasmic reticulum kinase(PERK),activating transcription factor 6(ATF6)and inositol-requiring enzyme 1(IRE1)and the related downstream signals(C/EBP-homologous protein(CHOP)and Caspase 12)were activated,as well as the ER-resident chaperone Glucose-regulated protein 78(GRP78).In addition,VP1 upregulated the autophagy marker Microtubule-associated protein 1 light chain 3 beta(LC3B),while PMP22 silencing or gp78 overexpression reversed the phenomenon.Meanwhile,PMP22 silencing or gp78 overexpression increased proliferation of EV71-VP1-transfected mouse SCs.Conclusion:Gp78 could regulate PMP22 accumulation through ubiquitination degradation and cause ER stress and autophagy in EV71-VP1-overexpressing mouse SCs.Therefore,the gp78/PMP22/ER stress axis might emerge as a promising therapeutic target for myelin and neuronal damage induced by EV71 infection.展开更多
Human enterovirus 71(EV71)is the main causative pathogen of hand,foot,and mouth disease(HFMD)in children.The epidemic of HFMD has been a public health problem in Asia-Pacific region for decades,and no vaccine and effe...Human enterovirus 71(EV71)is the main causative pathogen of hand,foot,and mouth disease(HFMD)in children.The epidemic of HFMD has been a public health problem in Asia-Pacific region for decades,and no vaccine and effective antiviral medicine are available.Curcumin has been used as a traditional medicine for centuries to treat a diversity of disorders including viral infections.In this study,we demonstrated that curcumin showed potent antiviral effect again EV71.In Vero cells infected with EV71,the addition of curcumin significantly suppressed the synthesis of viral RNA,the expression of viral protein,and the overall production of viral progeny.Similar with the previous reports,curcumin reduced the production of ROS induced by viral infection.However,the antioxidant property of curcumin did not contribute to its antiviral activity,since N-acetyl-L-cysteine,the potent antioxidant failed to suppress viral replication.This study also showed that extracellular signal-regulated kinase(ERK)was activated by either viral infection or curcumin treatment,but the activated ERK did not interfere with the antiviral effect of curcumin,indicating ERK is not involved in the antiviral mechanism of curcumin.Unlike the previous reports that curcumin inhibited protein degradation through ubiquitin–proteasome system(UPS),we found that curcumin had no impact on UPS in control cells.However,curcumin did reduce the activity of proteasomes which was increased by viral infection.In addition,the accumulation of the short-lived proteins,p53 and p21,was increased by the treatment of curcumin in EV71-infected cells.We further probed the antiviral mechanism of curcumin by examining the expression of GBF1 and PI4KB,both of which are required for the formation of viral replication complex.We found that curcumin significantly reduced the level of both proteins.Moreover,the decreased expression of either GBF1 or PI4KB by the application of siRNAs was sufficient to suppress viral replication.We also demonstrated that curcumin showed anti-apoptotic展开更多
文摘Hand foot and mouth disease is a febrile sickness complex characterized by cutaneous eruption (exanthem) on the palms and soles with simultaneous occurrence of muco-cutanous vesiculo-ulcerative lesions (enanthem) affecting the mouth. The illness is caused by a number of enteroviruses with coxsackievirus A16 and enterovirus 71 as the main causative agents. Human enterovirus 71 (EV71) belongs to the species Human enterovirus A under the genus Enterovirus within the family Picornaviridae. EV71 has been associated with an array of clinical diseases including hand foot and mouth disease (HFMD), aseptic meningitis, encephalitis and poliomyelitis-like acute flaccid paralysis. A large outbreak of HFMD due to highly neurovirulent EV71 emerged in Malaysia in 1997, and caused 41 deaths amongst young children. In late 2000, a recurrence of an outbreak of HFMD occurred in Malaysia with 8 fatalities in peninsular Malaysia. Outbreak of HFMD due to EV71 recurred in 2003 with an unknown number of cases and mortalities. A similar outbreak of HFMD with 2 recorded deaths in young children occurred in peninsular Malaysia in late 2005 and this was followed by a larger outbreak in Sarawak (Malaysian Borneo) with 6 reported fatalities in the early part of 2006. The current on-going outbreak of HFMD started in peninsular Malaysia in epidemiological week 12 of 2010. As with other HFMD outbreaks in Malaysia, both EV71 and CA16 were the main aetiological viruses isolated. In similarity with the HFMD outbreak in 2005, the isolation of CA16 preceded the appearance of EV71. Based on the VP1 gene nucleotide sequences, 4 sub-genogroups of EV71 (C1, C2, B3 and B4) co-circulated and caused the outbreak of hand, foot and mouth disease in peninsular Malaysia in 1997. Two sub-genogroups (C1 and B4) were noted to cause the outbreak in 2000 in both peninsular Malaysia and Sarawak. EV71 of sub-genogroup B5 with smaller contribution from sub-genogroup C1 caused the outbreak in 2003. In the 2005 outbreak, besides the EV71 strains of sub-genogroup C1, EV71
基金Supported by the National Science Council,No.93-2214-E-007-016 Ministry of Economic Affairs,No.93-EC-17A-17S1-0009
文摘AIM： Enterovirus 71 （EV71） has been implicated as the etiological agent responsible for the recent outbreaks of hand, foot and mouth disease associated with severe neurological diseases in the Asia-Pacific region. METHODS： The assembly process was hypothesized to occur via an orchestrated proteolytic processing of the P1 precursor by the viral protease 3CD. To test this hypothesis, we constructed 3 recombinant baculoviruses： Bac-P1 expressing P1; Bac-3CD expressing 3CD; and Bac-P1-3CD co-expressing P1 and 3CD. RESULTS： Both single infection by Bac-P1-3CD and coinfection by Bac-P1 and Bac-3CD resulted in correct cleavage of P1 to yield individual proteins VP0, VP1 and VP3, while the former approach yielded higher VLP production. In the cells, the structural proteins selfassembled into clusters of virus-like particles （VLP） resembling the authentic EV71 particle aggregates. After ultracentrifugation purification, the dispersed VLPs were indistinguishable from the authentic virus in size, appearance, composition and surface epitopes, as determined by SDS-PAGE, Western blot, transmission electron microscopy and immunogold labeling. CONCLUSION： Our data, for the first time, suggest that in insect cells EV71 structural proteins adopt a processing and assembly pathway similar to poliovirus assembly. The preservation of particle morphology and composition suggest that the VLP may be a valuable vaccine candidate to prevent EV71 epidemics.
文摘Background Enterovirus 71 （EV71） and coxsackievirus A16 （Cox A16） are major causative agents for hand, foot and mouth disease （HFMD）. Studies indicate that the frequent HFMD outbreaks result in a few hundreds children＇s death in China in recent years. The vaccine and other research for HFMD need to be developed urgently. The aims of our study were： to explore dynamic development of mother-source neutralizing antibodies against EV71 and Cox A16 in infants from Jiangsu Province, China, and to provide the fundamental data for further establishing of corresponding immunization course. Methods Peripheral blood samples were collected from 133 of parturient women once immediately before delivery and their infants at two and seven months of age. Method of micro-dose cytopathogenic effect was used to measure neutralizing antibodies against EV71 and Cox A16, respectively. Results Seropositive rates of anti-EV71 and anti-Cox A16 in prenatal women were 79.7% （106/133） and 92.5% （123/133）, respectively; geometric mean titers （GMTs） were 29.0 and 61.9; 75.9% （101/133） prenatal women were both positive in anti-EV71 and anti-Cox A16; seropositive rates of anti-EV71 and anti-Cox A16 were 25.6% （34/133） and 38.3% （51/133） in infants at two months of age; GMTs were 12.3 and 18.0, respectively. GMTs of anti-EV71 were significantly higher for infants at seven months （82.6） compared with that at two months （P 〈0.05）, showing infants had inapparently infected by EV71 during two to seven months. Although only one offspring （0.75%） at seven months was found having anti-Cox A16 transfered from maternal, this observation suggested no maternal antibody may remain in infants at seven months. Conclusions The prevalence of EV71 and Cox A16 were relatively high in Jiangsu Province. Bivalent vaccine against both EV71 and Cox A16 should be developed, and the ideal time point for prime immunization for infants is around 2-5 months of age.
基金This work was supported by the National Basic Research Program(973 Program)the National Programs for High Technology Research and Development Program(863 Program)(Grant Nos.2006CB806503,2006AA020502)+1 种基金National Major Project(Grant Nos.2009ZX10004-304,2009ZX09311-001)Tsinghua University Initiative Scientific Research Program(Grant No,2009THZ01).
文摘Enterovirus 71(EV71),one of the major causative agents for hand-foot-and-mouth disease(HFMD),has caused more than 100 deaths among Chinese children since March 2008.The EV71 genome encodes an RNAdependent RNA polymerase(RdRp),denoted 3D^(pol),which is central for viral genome replication and is a key target for the discovery of specific antiviral therapeutics.Here we report the crystal structures of EV71 RdRp(3D^(pol))and in complex with substrate guanosine-5'-triphosphate and analog 5-bromouridine-5'-triphosphate best to 2.4Åresolution.The structure of EV71 RdRp(3D^(pol))has a wider open thumb domain compared with the most closely related crystal structure of poliovirus RdRp.And the EV71 RdRp(3D^(pol))complex with GTP or Br-UTP bounded shows two distinct movements of the polymerase by substrate or analogue binding.The model of the complex with the template:primer derived by superimposition with foot-and-mouth disease virus(FMDV)3D/RNA complex reveals the likely recognition and binding of template:primer RNA by the polymerase.These results together provide a molecular basis for EV71 RNA replication and reveal a potential target for anti-EV71 drug discovery.
基金supported by the National Natural Science Foundation of China (Grant 81503118)CAMS Initiative for Innovative Medicine (CAMS-I2 M-1-010)The National Science and Technology Major Project of the Ministry of Science and Technology of China (2018ZX09711003-005-004)
文摘Enterovirus 71(EV71) is one of the main pathogens that causes hand-foot-and-mouth disease(HFMD). HFMD caused by EV71 infection is mostly self-limited; however, some infections can cause severe neurological diseases, such as aseptic meningitis, brain stem encephalitis, and even death. There are still no effective clinical drugs used for the prevention and treatment of HFMD. Studying EV71 protein function is essential for elucidating the EV71 replication process and developing anti-EV71 drugs and vaccines. In this review, we summarized the recent progress in the studies of EV71 noncoding regions(50 UTR and 30 UTR) and all structural and nonstructural proteins, especially the key motifs involving in viral infection, replication, and immune regulation. This review will promote our understanding of EV71 virus replication and pathogenesis, and will facilitate the development of novel drugs or vaccines to treat EV71.
基金National Science and Technology Major Project (2008ZX10004-014)
文摘The comparative analysis of the biological characterization and the genetic background study of EV71 circulating strains is commonly recognized as basic work necessary for development of an effective EV71 vaccine. In this study, we sequenced five EV71 circulating strains, isolated from Fuyang, Hefei, Kunming and Shenzhen city of China and named them FY-23, FY-22, H44, K9 and S1 respectively. The sequence alignment demonstrated their genotypes be C4. The genetic distance of the VP1 gene from these isolates suggested that they were highly co-related with genetic identity similar to other previously reported EV71 strains in China. Additionally, these strains were identified to display some obvious proliferation dynamics and plaque morphology when propagated in Vero cells. However, a distinctive difference in pathogenic ability in neonatal mice was found. Some differences in cross neutralization test & immunogenic analysis were also found. All these results are related to the biological characterization of circulating EV71 strains in China and aid in the development of an EV71 vaccine in the future.
文摘Human enterovirus 71(HEV71) is the cause of hand,foot and mouth disease and associated neurological complications in children under five years of age.There has been an increase in HEV71 epidemic activity throughout the Asia-Pacific region in the past decade,and it is predicted to replace poliovirus as the extant neurotropic enterovirus of highest global public health significance. To date there is no effective antiviral treatment and no vaccine is available to prevent HEV71 infection. The increase in prevalence, virulence and geographic spread of HEV71 infection over the past decade provides increasing incentive for the development of new therapeutic and prevention strategies against this emerging viral infection. The current review focuses on the potential, advantages and disadvantages of these strategies. Since the explosion of outbreaks leading to large epidemics in China, research in natural therapeutic products has identified several groups of compounds with anti-HEV71 activities. Concurrently, the search for effective synthetic antivirals has produced promising results. Other therapeutic strategies including immunotherapy and the use of oligonucleotides have also been explored. A sound prevention strategy is crucial in order to control the spread of HEV71. To this end the ultimate goal is the rapid development, regulatory approval and widespread implementation of a safe and effective vaccine. The various forms of HEV71 vaccine designs are highlighted in this review. Given the rapid progress of research in this area, eradication of the virus is likely to be achieved.
文摘A sensitive reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for human enterovirus 71 (EV71) and Coxsackievirus A16 (CVA16) infection was further evaluated. The one step reaction was performed in a single tube at 65?C for 45 min for EV71 and 35 min for CVA16. The detection limits of RT-LAMP assays for both EV71 and CVA16 were 0.1 of a 50% tissue culture infective dose (TCID50) per reaction, based on 10—Fold dilutions of a titrated EV71 or CVA16 strain. The specific assay showed there were no cross-reactions with Coxsackievirus A (CVA) viruses (CVA 2, 4, 5, 7, 9, 10, 14, and 25), Coxsackievirus B (CVB) viruses (CVB 1, 2, 3, 4, and 5) or ECHO viruses (ECHO 3, 6, 11, and 19). In parallel with commercial quantitative real-time polymerase chain reaction (qRT-PCR) diagnostic kits for EV71 and CVA16, the RT-LAMP assay was evaluated with 515 clinical specimens, the results showed the RT-LAMP assay and the qRT-PCR assay were in complete agreement for 513/515 (99.6%) of the specimens. Two samples with discrepant results from two methods were further verified by nested reverse transcription polymerase chain reaction (nRT-PCR) assay and sequencing to be true positives for CVA16. In conclusion, RT-LAMP assay is demonstrated to be a sensitive and specific assay and have a great potential for the rapid and visual screening of EV71 and CVA16 in China, especially in those resource-limited hospitals and rural clinics of provincial and municipal regions.
文摘Human enterovirus 71 viruses have been long circulating throughout the world. In this study, we performed a positive selection analysis of the VP1 genes of capsid proteins from Enterovirus 71 viruses. Our results showed that although most sites were under negative or neutral evolution, four positions of the VP1 genes were under positive selection pressure. This might account for the spread and frequent outbreaks of the viruses and the enhanced neurovirulence. In particular, position 98 might be involved in neutralizing antibodies, modulating the virus-receptor interaction and enhancing the virulence of the viruses. Moreover, both positions 145 and 241 might correlate to determine the receptor specificity. However, these positions did not display much difference in amino acid polymorphism. In addition, no position in the VP1 genes of viruses isolated from China was under positive selection.
基金the Foreign Experts Bureau of Yunnan Province,Foreign Talents Program(2018,Grant No.YNZ2018002)Thailand Research grants entitled Biodiversity,phylogeny and role of fungal endophytes on above parts of Rhizophora apiculata and Nypa fruticans(Grant No.RSA5980068)+14 种基金the future of specialist fungi in a changing climate:baseline data for generalist and specialist fungi associated with ants,Rhododendron species and Dracaena species(Grant No.DBG6080013)Impact of climate change on fungal diversity and biogeography in the Greater Mekong Subregion(Grant No.RDG6130001)Chiang Mai University for the award of visiting ProfessorCAS President’s International Fellowship Initiative(PIFI)for funding his postdoctoral research(Grant No.2018PC0006)the National Science Foundation of China(NSFC,project code 31750110478)supported by the Graduate Program for the Undiscovered Taxa of Koreain part by the Project on Survey and Discovery of Indigenous Fungal Species of Korea funded by NIBR and Project on Discovery of Fungi from Freshwater and Collection of Fungarium funded by NNIBR of the Ministry of Environment(MOE)in part carried out with the support of Cooperative Research Program for Agriculture Science and Technology Development(PJ013744),Rural Development Administration,Republic of Koreain part supported by the BK21 plus program through the National Research Foundation(NRF)funded by the Ministry of Education of Korea.Jian-Kui Liu thanks the National Natural Science Foundation of China(NSFC 31600032)the CNPq(Conselho Nacional de Desenvolvimento Cientifico e Tecnologico)for a research grant(309058/2015-5)funding for collecting trips(401186/2014-8)a collaborative project with RL as Special Visiting Professor(314570/2014-4)Funding for phylogenetic work on Graphidaceae was provided by a grant from the National Science Foundation(NSF)to The Field Museum:DEB-1025861"ATM-Assembling a taxonomic monograph:The lichen family Graphidaceae"PI Thorsten Lumbsch,CoPI Robert Luckingthe CAPES,CNPq,and FAPEMIG for financial support and ICMBio
文摘This article is the tenth series of the Fungal Diversity Notes,where 114 taxa distributed in three phyla,ten classes,30 orders and 53 families are described and illustrated.Taxa described in the present study include one new family(viz.Pseudoberkleasmiaceae in Dothideomycetes),five new genera(Caatingomyces,Cryptoschizotrema,Neoacladium,Paramassaria and Trochilispora)and 71 new species,(viz.Acrogenospora thailandica,Amniculicola aquatica,A.guttulata,Angustimassarina sylvatica,Blackwellomyces lateris,Boubovia gelatinosa,Buellia viridula,Caatingomyces brasiliensis,Calophoma humuli,Camarosporidiella mori,Canalisporium dehongense,Cantharellus brunneopallidus,C.griseotinctus,Castanediella meliponae,Coprinopsis psammophila,Cordyceps succavus,Cortinarius minusculus,C.subscotoides,Diaporthe italiana,D.rumicicola,Diatrypella delonicis,Dictyocheirospora aquadulcis,D.taiwanense,Digitodesmium chiangmaiense,Distoseptispora dehongensis,D.palmarum,Dothiorella styphnolobii,Ellisembia aurea,Falciformispora aquatic,Fomitiporia carpinea,F.lagerstroemiae,Grammothele aurantiaca,G.micropora,Hermatomyces bauhiniae,Jahnula queenslandica,Kamalomyces mangrovei,Lecidella yunnanensis,Micarea squamulosa,Muriphaeosphaeria angustifoliae,Neoacladium indicum,Neodidymelliopsis sambuci,Neosetophoma miscanthi,N.salicis,Nodulosphaeria aquilegiae,N.thalictri,Paramassaria samaneae,Penicillium circulare,P.geumsanense,P.mali-pumilae,P.psychrotrophicum,P.wandoense,Phaeoisaria siamensis,Phaeopoacea asparagicola,Phaeosphaeria penniseti,Plectocarpon galapagoense,Porina sorediata,Pseudoberkleasmium chiangmaiense,Pyrenochaetopsis sinensis,Rhizophydium koreanum,Russula prasina,Sporoschisma chiangraiense,Stigmatomyces chamaemyiae,S.cocksii,S.papei,S.tschirnhausii,S.vikhrevii,Thysanorea uniseptata,Torula breviconidiophora,T.polyseptata,Trochilispora schefflerae and Vaginatispora palmae).Further,twelve new combinations(viz.Cryptoschizotrema cryptotrema,Prolixandromyces australi,P.elongatus,P.falcatus,P.longispinae,P.microveliae,P.neoalardi,P.polhemorum,P.protubera
基金supported by grants from the Health Science and Technology Program of Hangzhou(No.2011A039)the Medical Science and Technology Program of Zhejiang Province(No.2012KYB159).
文摘Background:Hand-foot-and-mouth disease(HFMD)is a common pediatric infectious disease caused by a variety of intestinal viruses.Enterovirus 71(EV71)is the primary pathogen that might cause severe symptoms and even death in children with HFMD.This study aimed to investigate the intestinal detoxification time of HFMD children with EV71 infection and its related factors.Methods:Sixty-five HFMD children with EV71 infection were followed up.Their stool samples were collected once every 4 to 7 days.Viral nucleic acids were detected byfl uorescent polymerase chain reaction until the results became negative.The positive rates of viral nucleic acids were analyzed by the Kaplan-Meier method.The Log-rank test and Cox-Mantel test were used to analyze factors affecting the HFMD children with EV71 infection.Results:On the 2nd,4th,6th and 10th week,the positive rates of viral nucleic acids in stool samples of the 65 children were 94.6%,48.1%,17.2%and 0,respectively.Univariate analysis showed that the intestinal detoxification time of the children were related to gender,pre-admission disease course,severity of disease,and use of steroids or gamma globulin(P<0.05).Multivariate analysis showed that the severity of disease was an independent factor affecting the intestinal detoxification time(P<0.05),with a relative risk of 2.418.Conclusions:The longest intestinal detoxification time of HFMD children with EV71 infection was 10 weeks.The severity of disease was an important factor affecting the intestinal detoxification time of HFMD children with EV71 infection.Severe HFMD children with EV71 infection had a longer intestinal detoxification time.
基金The study was supported by Guangdong Natural Science Foundation(Grant Numbers 2020A1515010014,2022A1515012411)Science and Technology Key Project for People’s Livelihood of Guangzhou,China(Grant Number 202206010060)+1 种基金Guangzhou Science and Technology Bureau Basic Research Project(SL2024A03J01288)Innovative Project of Children’s Research Institute,Guangzhou Women and Children’s Medical Center,China(Grant Numbers Pre-NSFC-2019-002,NKE PRE-2019-015).
文摘Background:During Enterovirus type 71(EV71)infection,the structural viral protein 1(VP1)activates endoplasmic reticulum(ER)stress associated with peripheral myelin protein 22(PMP22)accumulation and induces autophagy.However,the specific mechanism behind this process remains elusive.Methods:In this research,we used the VP1-overexpressing mouse Schwann cells(SCs)models co-transfected with a PMP22 silencing or Autocrine motility factor receptor(AMFR/gp78)overexpressing vector to explore the regulation of gp78 on PMP22 and its relationship with autophagy and apoptosis.Results:The activity of gp78 could be influenced by EV71-VP1,leading to a decrease in the ubiquitination and degradation of PMP22,resulting in PMP22 accumulation in ER.In VP1-overexpressing mouse SCs,all three ER stress sensors,including pancreatic endoplasmic reticulum kinase(PERK),activating transcription factor 6(ATF6)and inositol-requiring enzyme 1(IRE1)and the related downstream signals(C/EBP-homologous protein(CHOP)and Caspase 12)were activated,as well as the ER-resident chaperone Glucose-regulated protein 78(GRP78).In addition,VP1 upregulated the autophagy marker Microtubule-associated protein 1 light chain 3 beta(LC3B),while PMP22 silencing or gp78 overexpression reversed the phenomenon.Meanwhile,PMP22 silencing or gp78 overexpression increased proliferation of EV71-VP1-transfected mouse SCs.Conclusion:Gp78 could regulate PMP22 accumulation through ubiquitination degradation and cause ER stress and autophagy in EV71-VP1-overexpressing mouse SCs.Therefore,the gp78/PMP22/ER stress axis might emerge as a promising therapeutic target for myelin and neuronal damage induced by EV71 infection.
基金This work was supported by the grants of National Natural Science Foundation of China to Zhaohua Zhong(Grant No.81271825)Wenran Zhao(Grant No.31270198).
文摘Human enterovirus 71(EV71)is the main causative pathogen of hand,foot,and mouth disease(HFMD)in children.The epidemic of HFMD has been a public health problem in Asia-Pacific region for decades,and no vaccine and effective antiviral medicine are available.Curcumin has been used as a traditional medicine for centuries to treat a diversity of disorders including viral infections.In this study,we demonstrated that curcumin showed potent antiviral effect again EV71.In Vero cells infected with EV71,the addition of curcumin significantly suppressed the synthesis of viral RNA,the expression of viral protein,and the overall production of viral progeny.Similar with the previous reports,curcumin reduced the production of ROS induced by viral infection.However,the antioxidant property of curcumin did not contribute to its antiviral activity,since N-acetyl-L-cysteine,the potent antioxidant failed to suppress viral replication.This study also showed that extracellular signal-regulated kinase(ERK)was activated by either viral infection or curcumin treatment,but the activated ERK did not interfere with the antiviral effect of curcumin,indicating ERK is not involved in the antiviral mechanism of curcumin.Unlike the previous reports that curcumin inhibited protein degradation through ubiquitin–proteasome system(UPS),we found that curcumin had no impact on UPS in control cells.However,curcumin did reduce the activity of proteasomes which was increased by viral infection.In addition,the accumulation of the short-lived proteins,p53 and p21,was increased by the treatment of curcumin in EV71-infected cells.We further probed the antiviral mechanism of curcumin by examining the expression of GBF1 and PI4KB,both of which are required for the formation of viral replication complex.We found that curcumin significantly reduced the level of both proteins.Moreover,the decreased expression of either GBF1 or PI4KB by the application of siRNAs was sufficient to suppress viral replication.We also demonstrated that curcumin showed anti-apoptotic
