The fragile antibody leads to a great challenge as a scaffold to fabricate the luminescent metal nanoclusters using one-pot method.This study presents a stable single-chain anti-body(scFv57R-ATS)for the fabrication of...The fragile antibody leads to a great challenge as a scaffold to fabricate the luminescent metal nanoclusters using one-pot method.This study presents a stable single-chain anti-body(scFv57R-ATS)for the fabrication of luminescent gold nanoclusters(AuNCs@scFv57R-ATS)and a quick,sensitive rabies virus detection in living cells.In this paper,AuNCs@scFv57R-ATS was designed to specifically recognize antigen RV in modified HeLa cells,which promoted the demonstration of metal nanocluster fluorescent probes for antigen targeting and therapy.展开更多
猪塞内加谷病毒(Seneca Valley virus,SVV)是一种新出现的可感染仔猪及母猪并导致仔猪死亡的病毒。2015年3月份至今年在我国、巴西及美国出现了几起塞内加谷病毒感染猪群并伴随严重的临床症状及发病死亡的疫情,造成严重的经济损失。为...猪塞内加谷病毒(Seneca Valley virus,SVV)是一种新出现的可感染仔猪及母猪并导致仔猪死亡的病毒。2015年3月份至今年在我国、巴西及美国出现了几起塞内加谷病毒感染猪群并伴随严重的临床症状及发病死亡的疫情,造成严重的经济损失。为分离鉴定SVV,本研究将RT-PCR检测为SVV阳性的猪临床样品无菌处理后接种PK-15细胞,连续传代培养。通过细胞病变、RT-PCR扩增、电镜观察和基因序列测定对细胞培养物进行鉴定。结果表明我们成功分离到国内首株SVV,并将其命名为SVV CH-01-2015。全基因组的系统发育分析表明SVV CH-01-2015位于Senecavirus病毒属,与Senecavirus病毒属中的病毒成员同源性最高,由此可以确定本研究所分离到的SVV属于Senecavirus病毒属。本研究为进一步研究SVV的致病性和致病机制奠定了基础。展开更多
番茄褐色皱果病毒Tomato brown rugose fruit virus(ToBRFV)于2014年首次在以色列发现,随后传播到欧洲、美洲以及亚洲等地。ToBRFV在番茄叶片上引起花叶,更重要的是在番茄果实上引起褐色皱缩斑,导致番茄完全失去商品价值,是番茄安全生...番茄褐色皱果病毒Tomato brown rugose fruit virus(ToBRFV)于2014年首次在以色列发现,随后传播到欧洲、美洲以及亚洲等地。ToBRFV在番茄叶片上引起花叶,更重要的是在番茄果实上引起褐色皱缩斑,导致番茄完全失去商品价值,是番茄安全生产的重大威胁。为遏制ToBRFV的传播,多个国家已经将该病毒列入检疫对象。2019年,我们在山东番茄上检测到该病毒。本文综述了ToBRFV发生与危害、寄主范围和症状、传播方式、基因组结构、检测方法,并提出了防治建议,希望有助于防范该病毒在我国的扩散。展开更多
Chinese isolate of transmissible gastroenteritis virus(TGEV)was propagated and harvested in swine testicle(ST)cells.Two pairs of primers were designed according to the published sequence with Oligo 4.1 and DNasis soft...Chinese isolate of transmissible gastroenteritis virus(TGEV)was propagated and harvested in swine testicle(ST)cells.Two pairs of primers were designed according to the published sequence with Oligo 4.1 and DNasis softwares.The products of RT-PCR were named Sa and Sb,of 2.3kb and 2.1kb respectively.Sa was inserted in EcoR I and Kpn I sites after Sb was cloned in Kpn I and Pst I sites of the same pUC18 plasmid.The recombinant designated pUC-S was verified and analyzed by corresponding restriction endonuclease(RE)and nested PCR on the basis of genetic sites of S gene and physical map of pUC18 plasmid,which was identified as S gene from Chinese isolate of TGEV.展开更多
Accurate differentiation of the pathogenic phenotypes of infectious bursal disease viruses(IBDVs) will instruct effective vaccination programs and improve the study of the molecular epidemiology of IBDVs. In this stud...Accurate differentiation of the pathogenic phenotypes of infectious bursal disease viruses(IBDVs) will instruct effective vaccination programs and improve the study of the molecular epidemiology of IBDVs. In this study, an 833 bp hypervariable nucleotide region was identified in VP2 genes of known IBDVs with different virulences through multiple sequence alignment.Moreover, using NEBcutter software analysis, two restriction enzyme sites, SpeⅠ(generating 531 and 302 bp fragments) and StuⅠ(generating 242 and 591 bp fragments) were found presented in very virulent but not attenuated IBDVs. Moreover, the restriction enzyme site SacⅠ(generating 218 and 615 bp fragments) presented in attenuated IBDVs but not very virulent IBDVs. Therefore,a reverse-transcription(RT)-PCR combined with a restriction fragment length polymorphism(RFLP) assay was developed to differentiate attenuated and very virulent IBDVs. The RT-PCR assay was used to confirm 282 IBDV positive samples from 310 suspicious dead chicken samples. The 60 IBDV positive samples were used to evaluate the assay, followed by confirmation via gene sequencing and histopathological examinations of the bursas of Fabricius from chickens infected by these IBDVs. The results showed that 24 viral strains with SpeⅠand StuⅠsites were very virulent, causing severe pathological damage in the bursas of Fabricius, while36 viral strains with the SacⅠsite were attenuated IBDVs, exhibiting only slight pathological damage. The combined RT-PCR and RFLP assay provided a useful approach for differentiating the pathogenic phenotypes of IBDVs.展开更多
Porcine parvovirus(PPV) is one of the major causes of reproductive failure in pigs, which poses a great threat to the pig breeding industry and results in tremendous economic losses worldwide. Autophagy is the biologi...Porcine parvovirus(PPV) is one of the major causes of reproductive failure in pigs, which poses a great threat to the pig breeding industry and results in tremendous economic losses worldwide. Autophagy is the biological process of cell self-defense and self-protection. Despite many viruses can cause cell autophagy, when they enter cell or copied, the relationship between autophagy and PPV infection has not been reported. In this study, impact of autophagy after swine testicular(ST) cells infected by PPV was studied. Autophagy was demonstrated by the effective replication of PPV through transmission electron microscopy, immunofluorescence and western blot analysis. Moreover, autophagy was confirmed to benefit PPV replication by real-time fluorescence quantitative PCR and determination of median tissue culture infective dose(TCID). For the first time, the complex interaction between PPV infection and autophagy was explored in this study. It indicated that PPV could induce autophagy in ST cells, which in turn facilitated its own replication, which might be one of the mechanisms of the virus infection. These findings could facilitate the study of the pathogenesis of PPV infection and provide new insight into the development of effective therapeutic strategies.展开更多
Garlic virus infection is an important disease which affects garlic production,with the increasing years of planting,harm of virus is serious year by year,which seriously affect yield and quality of garlic.In order to...Garlic virus infection is an important disease which affects garlic production,with the increasing years of planting,harm of virus is serious year by year,which seriously affect yield and quality of garlic.In order to know the garlic virus effectively,the paper reviewed the research situation of several important garlic virus in virus species,origin,distribution,host range,symptom,route of transmission,classification,genome and detection technique and the prevention technology of garlic viruses.At the same ...展开更多
The objective of this study was to construct the infectious clone of encephalomyocarditis virus (EMCV) BJC3 strain.The genomic cDNA of the virus was amplified by three overlapping segments using RT-PCR,and cloned into...The objective of this study was to construct the infectious clone of encephalomyocarditis virus (EMCV) BJC3 strain.The genomic cDNA of the virus was amplified by three overlapping segments using RT-PCR,and cloned into low-copy plasmid pWSK29 to construct the full-length cDNA clone pWSKBJC3/ w.The pWSKBJC3/w was in vitro transcribed and transfected into BHK-21 cells to rescue the virus.The results showed that the full-length cDNA clone was infectious and the virus could be rescued in BHK-21 cells.The rescued virus designated RvBJC3W was identified by RT-PCR and indirect immunofluorescence assay(IFA).The rescued virus had similar growth characteristics to its parental virus BJC3 and retained pathogenicity for mice.Our results indicate that the first infectious cDNA clone of EMCV in China has been successfully established and provides an essential tool for investigating the molecular basis of pathogenicity of EMCV.展开更多
OBJECTIVE How infection of Herpes simplex virus typeⅠ(HSV-1) induces enhancement of autophagy.MEHTODS The wild type HSV-1 strain Kos 1.1 was propagated in Vero cells and purified.SK-N-SH cells seeded in DMEM/F12 were...OBJECTIVE How infection of Herpes simplex virus typeⅠ(HSV-1) induces enhancement of autophagy.MEHTODS The wild type HSV-1 strain Kos 1.1 was propagated in Vero cells and purified.SK-N-SH cells seeded in DMEM/F12 were exposed to HSV-1 with 6 h or 12 h and multiplicity of infection(MOI) for 10 or 40 in each experiment.The infectious titers of the HSV-1 samples were determined by plaque assays.MDC staining to test the number of autophagosome within the cell after infection with time and moi was indicated in each experiment.At the molecular level,Western blotting and immunofluorescence analyses were done to study the expression of the proteins related to the cell autophagy.The mRNA transcribed from the gene related to autophagy was quantified by reverse transcription followed by real-time PCR.After intranasal infection of different transgenic mice,immunoflurorence studies were done to detected the expression of Aβ42 and proteins related to autophagy from the brain sections.Morris water maze experiment was performed to test the change of spatial learning and memory between different transgenic mice.RESULTS SK-N-SH cell showed time-and moi-dependent increase of MDC positive staining after HSV-1 infection.Western blotting analysis showed that LC3-Ⅱ was less in mock-infected cells but it was detected after 12 h from 10 to 40 moi HSV-1 infected cells.The level increased in a viral concentration-dependent manner.In agreement with the Western blotting results,direct fluorescence microscopy revealed that the signals of LC3 were consistent with their localization on autophagic compartments.P62,another protein related to autophagoysome formation,also increased with MOI.15 ku fragment of intracellular apolipoproteins E(APOE) protein increased after infection,but at the mRNA level it remained the same.The expression of APP showed less decrease but intracellular Aβ42 increased significantly compared with the mock group.Within the brain,after intranasal infection for 7 d,autophagy related proteins LC3 b and P62 increased as well,at the same time Aβ42 was found co-localized with LC3 b within the cell.Behavior test revealed that 17-month-old APOE4 mice had pool spatial learning and memory after infection compared with other groups.CONCLUSION HSV-1 induces an autophagic response and accelerates the fragmentation of APOE protein.展开更多
There are a total of more than 40 reported maize viral diseases worldwide. Five of them have reportedly occurred in China. They are maize rough dwarf disease, maize dwarf mosaic disease, maize streak dwarf disease, ma...There are a total of more than 40 reported maize viral diseases worldwide. Five of them have reportedly occurred in China. They are maize rough dwarf disease, maize dwarf mosaic disease, maize streak dwarf disease, maize crimson leaf disease, maize wallaby ear disease and corn lethal necrosis disease. This paper reviewed their occurrence and distribution as well as virus identification techniques in order to provide a basis for virus identification and diagnosis in corn production.展开更多
Objective:It intended to examine whether there is BDV infection in the human tumor tissues of central nervous system in China and investigate the correlation between BDV infection and tumors of central nervous system....Objective:It intended to examine whether there is BDV infection in the human tumor tissues of central nervous system in China and investigate the correlation between BDV infection and tumors of central nervous system.Methods:Nested reverse transcriptase polymerase chain reaction(nRT-PCR)and fluorescence quantitative polymerase chain reaction(FQ-PCR)was used to detect the BDV p24 fragments in 60 samples of human tumor tissues of central nervous system and 14 normal brain tissues.Results:The study indicated the positive rate of the BDV p24 fragment in human tumor tissues of the central nervous system(6.67%)was higher than that in normal brain tissues(0),but no statistical significance(P>0.05).Conclusion:It suggests that the BDV infection is present in the human tumor tissues of central nervous system in China, while the sample size wasn't large enough and we could not certify the possible correlation between BDV infection and cenfral nervous system tumors.展开更多
文摘The fragile antibody leads to a great challenge as a scaffold to fabricate the luminescent metal nanoclusters using one-pot method.This study presents a stable single-chain anti-body(scFv57R-ATS)for the fabrication of luminescent gold nanoclusters(AuNCs@scFv57R-ATS)and a quick,sensitive rabies virus detection in living cells.In this paper,AuNCs@scFv57R-ATS was designed to specifically recognize antigen RV in modified HeLa cells,which promoted the demonstration of metal nanocluster fluorescent probes for antigen targeting and therapy.
文摘猪塞内加谷病毒(Seneca Valley virus,SVV)是一种新出现的可感染仔猪及母猪并导致仔猪死亡的病毒。2015年3月份至今年在我国、巴西及美国出现了几起塞内加谷病毒感染猪群并伴随严重的临床症状及发病死亡的疫情,造成严重的经济损失。为分离鉴定SVV,本研究将RT-PCR检测为SVV阳性的猪临床样品无菌处理后接种PK-15细胞,连续传代培养。通过细胞病变、RT-PCR扩增、电镜观察和基因序列测定对细胞培养物进行鉴定。结果表明我们成功分离到国内首株SVV,并将其命名为SVV CH-01-2015。全基因组的系统发育分析表明SVV CH-01-2015位于Senecavirus病毒属,与Senecavirus病毒属中的病毒成员同源性最高,由此可以确定本研究所分离到的SVV属于Senecavirus病毒属。本研究为进一步研究SVV的致病性和致病机制奠定了基础。
文摘番茄褐色皱果病毒Tomato brown rugose fruit virus(ToBRFV)于2014年首次在以色列发现,随后传播到欧洲、美洲以及亚洲等地。ToBRFV在番茄叶片上引起花叶,更重要的是在番茄果实上引起褐色皱缩斑,导致番茄完全失去商品价值,是番茄安全生产的重大威胁。为遏制ToBRFV的传播,多个国家已经将该病毒列入检疫对象。2019年,我们在山东番茄上检测到该病毒。本文综述了ToBRFV发生与危害、寄主范围和症状、传播方式、基因组结构、检测方法,并提出了防治建议,希望有助于防范该病毒在我国的扩散。
文摘Chinese isolate of transmissible gastroenteritis virus(TGEV)was propagated and harvested in swine testicle(ST)cells.Two pairs of primers were designed according to the published sequence with Oligo 4.1 and DNasis softwares.The products of RT-PCR were named Sa and Sb,of 2.3kb and 2.1kb respectively.Sa was inserted in EcoR I and Kpn I sites after Sb was cloned in Kpn I and Pst I sites of the same pUC18 plasmid.The recombinant designated pUC-S was verified and analyzed by corresponding restriction endonuclease(RE)and nested PCR on the basis of genetic sites of S gene and physical map of pUC18 plasmid,which was identified as S gene from Chinese isolate of TGEV.
基金Supported by the National Technology and Research Project of China(2015BAD12B01-4)
文摘Accurate differentiation of the pathogenic phenotypes of infectious bursal disease viruses(IBDVs) will instruct effective vaccination programs and improve the study of the molecular epidemiology of IBDVs. In this study, an 833 bp hypervariable nucleotide region was identified in VP2 genes of known IBDVs with different virulences through multiple sequence alignment.Moreover, using NEBcutter software analysis, two restriction enzyme sites, SpeⅠ(generating 531 and 302 bp fragments) and StuⅠ(generating 242 and 591 bp fragments) were found presented in very virulent but not attenuated IBDVs. Moreover, the restriction enzyme site SacⅠ(generating 218 and 615 bp fragments) presented in attenuated IBDVs but not very virulent IBDVs. Therefore,a reverse-transcription(RT)-PCR combined with a restriction fragment length polymorphism(RFLP) assay was developed to differentiate attenuated and very virulent IBDVs. The RT-PCR assay was used to confirm 282 IBDV positive samples from 310 suspicious dead chicken samples. The 60 IBDV positive samples were used to evaluate the assay, followed by confirmation via gene sequencing and histopathological examinations of the bursas of Fabricius from chickens infected by these IBDVs. The results showed that 24 viral strains with SpeⅠand StuⅠsites were very virulent, causing severe pathological damage in the bursas of Fabricius, while36 viral strains with the SacⅠsite were attenuated IBDVs, exhibiting only slight pathological damage. The combined RT-PCR and RFLP assay provided a useful approach for differentiating the pathogenic phenotypes of IBDVs.
基金Supported by the National Natural Science Foundation of China(31372438,31201911)
文摘Porcine parvovirus(PPV) is one of the major causes of reproductive failure in pigs, which poses a great threat to the pig breeding industry and results in tremendous economic losses worldwide. Autophagy is the biological process of cell self-defense and self-protection. Despite many viruses can cause cell autophagy, when they enter cell or copied, the relationship between autophagy and PPV infection has not been reported. In this study, impact of autophagy after swine testicular(ST) cells infected by PPV was studied. Autophagy was demonstrated by the effective replication of PPV through transmission electron microscopy, immunofluorescence and western blot analysis. Moreover, autophagy was confirmed to benefit PPV replication by real-time fluorescence quantitative PCR and determination of median tissue culture infective dose(TCID). For the first time, the complex interaction between PPV infection and autophagy was explored in this study. It indicated that PPV could induce autophagy in ST cells, which in turn facilitated its own replication, which might be one of the mechanisms of the virus infection. These findings could facilitate the study of the pathogenesis of PPV infection and provide new insight into the development of effective therapeutic strategies.
基金Supported by Heilongjiang Natural Science Fund Item (C2004-05)
文摘Garlic virus infection is an important disease which affects garlic production,with the increasing years of planting,harm of virus is serious year by year,which seriously affect yield and quality of garlic.In order to know the garlic virus effectively,the paper reviewed the research situation of several important garlic virus in virus species,origin,distribution,host range,symptom,route of transmission,classification,genome and detection technique and the prevention technology of garlic viruses.At the same ...
基金supported by the Key Project of National Natural Science Foundation of China(30530550)
文摘The objective of this study was to construct the infectious clone of encephalomyocarditis virus (EMCV) BJC3 strain.The genomic cDNA of the virus was amplified by three overlapping segments using RT-PCR,and cloned into low-copy plasmid pWSK29 to construct the full-length cDNA clone pWSKBJC3/ w.The pWSKBJC3/w was in vitro transcribed and transfected into BHK-21 cells to rescue the virus.The results showed that the full-length cDNA clone was infectious and the virus could be rescued in BHK-21 cells.The rescued virus designated RvBJC3W was identified by RT-PCR and indirect immunofluorescence assay(IFA).The rescued virus had similar growth characteristics to its parental virus BJC3 and retained pathogenicity for mice.Our results indicate that the first infectious cDNA clone of EMCV in China has been successfully established and provides an essential tool for investigating the molecular basis of pathogenicity of EMCV.
基金National Natural Science Fundation of China (81471232).
文摘OBJECTIVE How infection of Herpes simplex virus typeⅠ(HSV-1) induces enhancement of autophagy.MEHTODS The wild type HSV-1 strain Kos 1.1 was propagated in Vero cells and purified.SK-N-SH cells seeded in DMEM/F12 were exposed to HSV-1 with 6 h or 12 h and multiplicity of infection(MOI) for 10 or 40 in each experiment.The infectious titers of the HSV-1 samples were determined by plaque assays.MDC staining to test the number of autophagosome within the cell after infection with time and moi was indicated in each experiment.At the molecular level,Western blotting and immunofluorescence analyses were done to study the expression of the proteins related to the cell autophagy.The mRNA transcribed from the gene related to autophagy was quantified by reverse transcription followed by real-time PCR.After intranasal infection of different transgenic mice,immunoflurorence studies were done to detected the expression of Aβ42 and proteins related to autophagy from the brain sections.Morris water maze experiment was performed to test the change of spatial learning and memory between different transgenic mice.RESULTS SK-N-SH cell showed time-and moi-dependent increase of MDC positive staining after HSV-1 infection.Western blotting analysis showed that LC3-Ⅱ was less in mock-infected cells but it was detected after 12 h from 10 to 40 moi HSV-1 infected cells.The level increased in a viral concentration-dependent manner.In agreement with the Western blotting results,direct fluorescence microscopy revealed that the signals of LC3 were consistent with their localization on autophagic compartments.P62,another protein related to autophagoysome formation,also increased with MOI.15 ku fragment of intracellular apolipoproteins E(APOE) protein increased after infection,but at the mRNA level it remained the same.The expression of APP showed less decrease but intracellular Aβ42 increased significantly compared with the mock group.Within the brain,after intranasal infection for 7 d,autophagy related proteins LC3 b and P62 increased as well,at the same time Aβ42 was found co-localized with LC3 b within the cell.Behavior test revealed that 17-month-old APOE4 mice had pool spatial learning and memory after infection compared with other groups.CONCLUSION HSV-1 induces an autophagic response and accelerates the fragmentation of APOE protein.
基金Supported by the Finance Department of Hebei Province(A2012120104)
文摘There are a total of more than 40 reported maize viral diseases worldwide. Five of them have reportedly occurred in China. They are maize rough dwarf disease, maize dwarf mosaic disease, maize streak dwarf disease, maize crimson leaf disease, maize wallaby ear disease and corn lethal necrosis disease. This paper reviewed their occurrence and distribution as well as virus identification techniques in order to provide a basis for virus identification and diagnosis in corn production.
文摘Objective:It intended to examine whether there is BDV infection in the human tumor tissues of central nervous system in China and investigate the correlation between BDV infection and tumors of central nervous system.Methods:Nested reverse transcriptase polymerase chain reaction(nRT-PCR)and fluorescence quantitative polymerase chain reaction(FQ-PCR)was used to detect the BDV p24 fragments in 60 samples of human tumor tissues of central nervous system and 14 normal brain tissues.Results:The study indicated the positive rate of the BDV p24 fragment in human tumor tissues of the central nervous system(6.67%)was higher than that in normal brain tissues(0),but no statistical significance(P>0.05).Conclusion:It suggests that the BDV infection is present in the human tumor tissues of central nervous system in China, while the sample size wasn't large enough and we could not certify the possible correlation between BDV infection and cenfral nervous system tumors.
