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
Human caring is regarded as the essence and core of nursing.Based on Dr.Watson′s human caring science theory,nursing staff in Wuhan Union Hospital and other healthcare institutions have delivered warm human caring se...Human caring is regarded as the essence and core of nursing.Based on Dr.Watson′s human caring science theory,nursing staff in Wuhan Union Hospital and other healthcare institutions have delivered warm human caring service in patients′ daily care.During the outbreak of major infectious disease-pandemic of COVID-19,patients are suffering from great psychological distress and pressure.Like nursing professionals around the world,nurses in Wuhan,Hubei,China,along with the colleagues from the National Aid Medical Team,used entire of themselves to conduct diversity human caring measures to patients suffering from the major infectious disease with love and responsibilities,which greatly demonstrated the essence and value of nursing care,and has wide implications for further development of nursing.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
Preliminary study has been made to test wether three strains McAb（1B1,5D<sub>6</sub>, 6D<sub>8</sub>）are against the same antigen determinant.Througn ELISA additive and competition expeiments...Preliminary study has been made to test wether three strains McAb（1B1,5D<sub>6</sub>, 6D<sub>8</sub>）are against the same antigen determinant.Througn ELISA additive and competition expeiments,itproved that these three strains are against different antigen determinant.The result of positiveserum antigen component analysis with 2 strains IBDV McAb showed that sample IBD positiveserum had obvious inhibition against combination of 5D<sub>6</sub> McAb with corresponding antigen.Theresults of substitution of corresponding component in ELISA inhibition experiment and compari-son of non-IBD serum （SPF chicken serum,ND,MD,IA positive serum）proved that IBD anti-serum was the only one showing inhibition against 5D<sub>6</sub> McAb.Comparison with AGP and electro-microscope observation showed that ELISA inhibition experiment was characterised by high-specificity,rapidity and sensitiveness.799 serum samples were tested with ELISA inhibition anddouble immunodiffusion（AGP） experiments.ELISA had gotten 486 positive,with positive rate of60.83%;and AGP 334。展开更多
In soybean cotyledonary transformation,Agrobacterium infection is influenced by many factors.The cultivation stage of Agrobacterium strain and infectious concentration were studied in this research.Results showed that...In soybean cotyledonary transformation,Agrobacterium infection is influenced by many factors.The cultivation stage of Agrobacterium strain and infectious concentration were studied in this research.Results showed that the highest rate of resistant shoots was obtained when the strain cultivation concentration was OD 600 =0.6,and the infectious concentration was OD 600 =0.5.Seven soybean cultivars in North China were also screened for high susceptibility to Agrobacterium and three genotypes were found to be s...展开更多
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.
文摘Human caring is regarded as the essence and core of nursing.Based on Dr.Watson′s human caring science theory,nursing staff in Wuhan Union Hospital and other healthcare institutions have delivered warm human caring service in patients′ daily care.During the outbreak of major infectious disease-pandemic of COVID-19,patients are suffering from great psychological distress and pressure.Like nursing professionals around the world,nurses in Wuhan,Hubei,China,along with the colleagues from the National Aid Medical Team,used entire of themselves to conduct diversity human caring measures to patients suffering from the major infectious disease with love and responsibilities,which greatly demonstrated the essence and value of nursing care,and has wide implications for further development of nursing.
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.
文摘Preliminary study has been made to test wether three strains McAb（1B1,5D<sub>6</sub>, 6D<sub>8</sub>）are against the same antigen determinant.Througn ELISA additive and competition expeiments,itproved that these three strains are against different antigen determinant.The result of positiveserum antigen component analysis with 2 strains IBDV McAb showed that sample IBD positiveserum had obvious inhibition against combination of 5D<sub>6</sub> McAb with corresponding antigen.Theresults of substitution of corresponding component in ELISA inhibition experiment and compari-son of non-IBD serum （SPF chicken serum,ND,MD,IA positive serum）proved that IBD anti-serum was the only one showing inhibition against 5D<sub>6</sub> McAb.Comparison with AGP and electro-microscope observation showed that ELISA inhibition experiment was characterised by high-specificity,rapidity and sensitiveness.799 serum samples were tested with ELISA inhibition anddouble immunodiffusion（AGP） experiments.ELISA had gotten 486 positive,with positive rate of60.83%;and AGP 334。
基金Supported by New Variety Breeding Key Project of Genetic Modified Organisms (2008ZX08004-002)Special Research Fund from Harbin Technology and Science Innovation (2008RFQXN013)Postdoctoral Fund and Doctoral Fund of Northeast Agricultural University
文摘In soybean cotyledonary transformation,Agrobacterium infection is influenced by many factors.The cultivation stage of Agrobacterium strain and infectious concentration were studied in this research.Results showed that the highest rate of resistant shoots was obtained when the strain cultivation concentration was OD 600 =0.6,and the infectious concentration was OD 600 =0.5.Seven soybean cultivars in North China were also screened for high susceptibility to Agrobacterium and three genotypes were found to be s...
