The complete genomic sequence of Duck hepatitis virus 1(DHV-1) ZJ-V isolate was sequenced and determined to be 7 691 nucleotides(nt) in length with a 5'-terminal un-translated region(UTR) of 626 nt and a 3'-te...The complete genomic sequence of Duck hepatitis virus 1(DHV-1) ZJ-V isolate was sequenced and determined to be 7 691 nucleotides(nt) in length with a 5'-terminal un-translated region(UTR) of 626 nt and a 3'-terminal UTR of 315 nt(not including the poly(A) tail).One large open reading frame(ORF) was found within the genome(nt 627 to 7 373) coding for a polypeptide of 2 249 amino acids.Our data also showed that the poly(A) tail of DHV-1 has at least 22 A's.Sequence comparison revealed significant homology(from 91.9% to 95.7%) between the protein sequences of the ZJ-V isolate and those of 21 reference isolates.Although DHV-1 has been classified as an unassigned virus in the Picornaviridae family,its genome showed some unique characteristics.DHV-1 contains 3 copies of the 2A gene and only 1 copy of the 3B gene,and its 3'-NCR is longer than those of other picornaviruses.Phylogenetic analysis to do sequence homology based on the VP1 protein sequences showed that the ZJ-V isolate shares high sequence homology with the reported DHV-1 isolates(from 92.9% to 99.2%),indicating that DHV-1 is genetically stable.展开更多
AIM: To establish a cell model harboring replicative clinical hepatitis B virus (HBV) isolates and evaluate its application in individualized selection of anti-HBV agents for chronic hepatitis B (CHB) patients. METHOD...AIM: To establish a cell model harboring replicative clinical hepatitis B virus (HBV) isolates and evaluate its application in individualized selection of anti-HBV agents for chronic hepatitis B (CHB) patients. METHODS: The full-length HBV genomic DNA from 8 CHB patients was amplified by polymerase chain reaction (PCR). All the patients were treated with lamivudine for at least seven months and finally became resistant to lamivudine. The amplified HBV DNA fragments were inserted into pHY106 vectors by Sap Ⅰ?digestion. The recombinant plasmids containing 1.1 copies of HBV genome were transiently transfected into Huh7 cell line, and the levels of HBsAg, HBeAg and intercellular HBV replicative intermediates were determined by ELISA and Southern blot analysis, respectively, with or without lamivudine and adefovir treatment. The antiviral treatment with adefovir was administered to the patients and analyzed in parallel. RESULTS: A total of 25 independent HBV isolateswere obtained from the sera of 8 patients, each patient had at least two isolates. One isolate from each individual was selected and subcloned into pHY106 vector, including 5 isolates with YVDD mutation and 3 isolates with YIDD mutation. All recombinant plasmids harboring HBV isolates were transfected into Huh7 cells. The results indicated that HBV genome carried in HBV replicons of clinical HBV isolates could effectively replicate and express in Huh7 cells. Adefovir, but not lamivudine, inhibited HBV replication both in vitro and in vivo, and in vitro inhibition was dose-dependent. CONCLUSION: The novel method described herein enables individualized selection of anti-HBV agents in clinic and is useful in future studies of antiviral therapy for CHB.展开更多
Background: Although hepatitis C virus (HCV) transmission through tissue transplantation has been rarely reported, a donor with undetected viremia may infect several recipients. A patient developed acute hepatitis C s...Background: Although hepatitis C virus (HCV) transmission through tissue transplantation has been rarely reported, a donor with undetected viremia may infect several recipients. A patient developed acute hepatitis C shortly after tissue transplantation. Ninety-one tissues or organs had been recovered from the donor. Objective: To determine whether the donor was the source of infection and the extent of transmission to other organ and tissue recipients. Design: Descriptive epidemiologic study; serum testing for HCV infection. Setting: Recipients were located in 16 states and 2 other countries. Participants: Donor and graft recipients. Measurements: Hepatitis C virus infection was defined as the presence of anti-HCV or HCV RNA. The authors determined the genetic relatedness of viral isolates from the donor and recipients by genotype comparison and quasi-species analysis. Results: The donor was anti-HCV-negative but was HCV RNA-positive (genotype 1a). Forty persons received transplants during 22 months. Five persons were HCV-infected before transplantation or had a genotype other than 1a, and 5 persons had no post-transplantation serum specimens available. Of the remaining 30 recipients, HCV infection occurred in 8 recipients: 3 of 3 organ recipients, 1 of 2 saphenous vein recipients, 1 of 3 tendon recipients, and 3 of 3 tendon with bone recipients. These 8 recipients had viral isolates genetically related to those of the donor. No cases occurred in recipients of skin (n = 2), cornea (n = 1), or irradiated bone (n = 16). Limitations: Post-transplantation serum specimens were unavailable for 5 recipients. Conclusions: An anti-HCV-negative donor was the source of HCV infection for 8 recipients of organs or tissues. Although HCV transmission from anti-HCV-negative donors is probably uncommon, changes in donor screening to include routine testing for HCV RNA merit further consideration to improve the safety of transplantation.展开更多
Genetic diversity of 51 oil-tea camellia germplasms was analyzed using the optimized inter-simple sequence repeat （ISSR）-PCR reaction system with 22 primers screened from a set of 100 ISSR primers. The results showe...Genetic diversity of 51 oil-tea camellia germplasms was analyzed using the optimized inter-simple sequence repeat （ISSR）-PCR reaction system with 22 primers screened from a set of 100 ISSR primers. The results showed that 493 discernible loci with distinct elec- trophoretic bands were obtained, of which, 478 loci （96.78%） were polymorphic. This indicated that oil-tea germplasms possess abundant genetic diversities. By clustering analysis performed using softwares of NTSYS 2.10 and Winboot, 51 oil-tea germplasms were divided into two groups： Group I had 48 lines of Camellia oleifera Abel, while Group II had three C. oleifera Abel related species and their similarity coefficient was 0.62. Group I was further divided into Group I-1 and Group I-2, and their similarity coefficient （Gs） was 0.634. All members of Group I-1 originated from Hunan Province, while Group I-2 included the rest of Hunan lines and those originated from other regions of China. Analyzed by software POPGENE 1.32, the Shannon＇s information index （I＊） of genetic polymorphism was 0.3852, the genetic diversity among different region popula- tions （Ht） was 0.2537, the genetic diversity within populations （Hs） was 0.15545, the dif- ferentiation coefficient of genetic diversity among populations （Gst） was 0.3967, and the gene flow among populations （Nm＊） was 0.8262. The Nei＇s genetic distances between the Hunan population and the populations originated from other regions of China implied that geographic isolation strongly influenced genetic differentiation among populations. Meanwhile, seedling rootstock grafting and high grafting for tree crown produced genetic variations among clonal offsprings.展开更多
Strain T02-25 was selected from approximately 30 rhizosphere isolates of Trichoderma species isolated from roots of crops. Its biological activity against Rhizoctonia solani was determined for the control efficacy to ...Strain T02-25 was selected from approximately 30 rhizosphere isolates of Trichoderma species isolated from roots of crops. Its biological activity against Rhizoctonia solani was determined for the control efficacy to pepper seedling blight caused by R. solani in the field. The assay methods were treating R. solani sclerotia by Trichoderma conidial suspension (10^6cfu ml^-1) and scattering Thichoderma rice bran over the pepper root medium. The results showed that T02-25 was active against R.solani in both ways, and its control efficacy was 82.7% and 78.0%, respectively. In addition to comparison of the efficacy of the two application methods, the relationship of different factors in the control efficacy of Trichoderma against R. solani was discussed.展开更多
基金National Natural Science Foundation of China (30670074)
文摘The complete genomic sequence of Duck hepatitis virus 1(DHV-1) ZJ-V isolate was sequenced and determined to be 7 691 nucleotides(nt) in length with a 5'-terminal un-translated region(UTR) of 626 nt and a 3'-terminal UTR of 315 nt(not including the poly(A) tail).One large open reading frame(ORF) was found within the genome(nt 627 to 7 373) coding for a polypeptide of 2 249 amino acids.Our data also showed that the poly(A) tail of DHV-1 has at least 22 A's.Sequence comparison revealed significant homology(from 91.9% to 95.7%) between the protein sequences of the ZJ-V isolate and those of 21 reference isolates.Although DHV-1 has been classified as an unassigned virus in the Picornaviridae family,its genome showed some unique characteristics.DHV-1 contains 3 copies of the 2A gene and only 1 copy of the 3B gene,and its 3'-NCR is longer than those of other picornaviruses.Phylogenetic analysis to do sequence homology based on the VP1 protein sequences showed that the ZJ-V isolate shares high sequence homology with the reported DHV-1 isolates(from 92.9% to 99.2%),indicating that DHV-1 is genetically stable.
基金The National Natural Science Foundation of China, No. 30271170the Ph.D. Program Fund of Chinese Ministry of Education, No. 20070487152
文摘AIM: To establish a cell model harboring replicative clinical hepatitis B virus (HBV) isolates and evaluate its application in individualized selection of anti-HBV agents for chronic hepatitis B (CHB) patients. METHODS: The full-length HBV genomic DNA from 8 CHB patients was amplified by polymerase chain reaction (PCR). All the patients were treated with lamivudine for at least seven months and finally became resistant to lamivudine. The amplified HBV DNA fragments were inserted into pHY106 vectors by Sap Ⅰ?digestion. The recombinant plasmids containing 1.1 copies of HBV genome were transiently transfected into Huh7 cell line, and the levels of HBsAg, HBeAg and intercellular HBV replicative intermediates were determined by ELISA and Southern blot analysis, respectively, with or without lamivudine and adefovir treatment. The antiviral treatment with adefovir was administered to the patients and analyzed in parallel. RESULTS: A total of 25 independent HBV isolateswere obtained from the sera of 8 patients, each patient had at least two isolates. One isolate from each individual was selected and subcloned into pHY106 vector, including 5 isolates with YVDD mutation and 3 isolates with YIDD mutation. All recombinant plasmids harboring HBV isolates were transfected into Huh7 cells. The results indicated that HBV genome carried in HBV replicons of clinical HBV isolates could effectively replicate and express in Huh7 cells. Adefovir, but not lamivudine, inhibited HBV replication both in vitro and in vivo, and in vitro inhibition was dose-dependent. CONCLUSION: The novel method described herein enables individualized selection of anti-HBV agents in clinic and is useful in future studies of antiviral therapy for CHB.
文摘Background: Although hepatitis C virus (HCV) transmission through tissue transplantation has been rarely reported, a donor with undetected viremia may infect several recipients. A patient developed acute hepatitis C shortly after tissue transplantation. Ninety-one tissues or organs had been recovered from the donor. Objective: To determine whether the donor was the source of infection and the extent of transmission to other organ and tissue recipients. Design: Descriptive epidemiologic study; serum testing for HCV infection. Setting: Recipients were located in 16 states and 2 other countries. Participants: Donor and graft recipients. Measurements: Hepatitis C virus infection was defined as the presence of anti-HCV or HCV RNA. The authors determined the genetic relatedness of viral isolates from the donor and recipients by genotype comparison and quasi-species analysis. Results: The donor was anti-HCV-negative but was HCV RNA-positive (genotype 1a). Forty persons received transplants during 22 months. Five persons were HCV-infected before transplantation or had a genotype other than 1a, and 5 persons had no post-transplantation serum specimens available. Of the remaining 30 recipients, HCV infection occurred in 8 recipients: 3 of 3 organ recipients, 1 of 2 saphenous vein recipients, 1 of 3 tendon recipients, and 3 of 3 tendon with bone recipients. These 8 recipients had viral isolates genetically related to those of the donor. No cases occurred in recipients of skin (n = 2), cornea (n = 1), or irradiated bone (n = 16). Limitations: Post-transplantation serum specimens were unavailable for 5 recipients. Conclusions: An anti-HCV-negative donor was the source of HCV infection for 8 recipients of organs or tissues. Although HCV transmission from anti-HCV-negative donors is probably uncommon, changes in donor screening to include routine testing for HCV RNA merit further consideration to improve the safety of transplantation.
文摘Genetic diversity of 51 oil-tea camellia germplasms was analyzed using the optimized inter-simple sequence repeat （ISSR）-PCR reaction system with 22 primers screened from a set of 100 ISSR primers. The results showed that 493 discernible loci with distinct elec- trophoretic bands were obtained, of which, 478 loci （96.78%） were polymorphic. This indicated that oil-tea germplasms possess abundant genetic diversities. By clustering analysis performed using softwares of NTSYS 2.10 and Winboot, 51 oil-tea germplasms were divided into two groups： Group I had 48 lines of Camellia oleifera Abel, while Group II had three C. oleifera Abel related species and their similarity coefficient was 0.62. Group I was further divided into Group I-1 and Group I-2, and their similarity coefficient （Gs） was 0.634. All members of Group I-1 originated from Hunan Province, while Group I-2 included the rest of Hunan lines and those originated from other regions of China. Analyzed by software POPGENE 1.32, the Shannon＇s information index （I＊） of genetic polymorphism was 0.3852, the genetic diversity among different region popula- tions （Ht） was 0.2537, the genetic diversity within populations （Hs） was 0.15545, the dif- ferentiation coefficient of genetic diversity among populations （Gst） was 0.3967, and the gene flow among populations （Nm＊） was 0.8262. The Nei＇s genetic distances between the Hunan population and the populations originated from other regions of China implied that geographic isolation strongly influenced genetic differentiation among populations. Meanwhile, seedling rootstock grafting and high grafting for tree crown produced genetic variations among clonal offsprings.
文摘Strain T02-25 was selected from approximately 30 rhizosphere isolates of Trichoderma species isolated from roots of crops. Its biological activity against Rhizoctonia solani was determined for the control efficacy to pepper seedling blight caused by R. solani in the field. The assay methods were treating R. solani sclerotia by Trichoderma conidial suspension (10^6cfu ml^-1) and scattering Thichoderma rice bran over the pepper root medium. The results showed that T02-25 was active against R.solani in both ways, and its control efficacy was 82.7% and 78.0%, respectively. In addition to comparison of the efficacy of the two application methods, the relationship of different factors in the control efficacy of Trichoderma against R. solani was discussed.
