AIM: To develop a Fok-I nested polymerase chain reaction(PCR)-restriction fragment length polymorphism analysis(PRA) method for the detection of hepatitis B virus X region(HBx) V5 M mutation.METHODS: Nested PCR was ap...AIM: To develop a Fok-I nested polymerase chain reaction(PCR)-restriction fragment length polymorphism analysis(PRA) method for the detection of hepatitis B virus X region(HBx) V5 M mutation.METHODS: Nested PCR was applied into DNAs from 198 chronic patients at 2 different stages [121 patients with hepatocellular carcinoma(HCC) and 77 carrier patients]. To identify V5 M mutants, digestion of nested PCR amplicons by the restriction enzyme Fok-I(GGA TGN9↓) was done. For size comparison, the enzymetreated products were analyzed by electrophoresis on 2.5% agarose gels, stained with ethidium bromide, and visualized on a UV transilluminator.RESULTS: The assay enabled the identification of 69 patients(sensitivity of 34.8%; 46 HCC patients and 23 carrier patients). Our data also showed that V5 M prevalence in HCC patients was significantly higher than in carrier patients(47.8%, 22/46 patients vs 0%, 0/23 patients, P < 0.001), suggesting that HBx Ag V5 M mutation may play a pivotal role in HCC generation in chronic patients with genotype C infections.CONCLUSION: The Fok-I nested PRA developed in this study is a reliable and cost-effective method to detect HBx Ag V5 M mutation in chronic patients with genotype C2 infection.展开更多
Microstructure of the deepest zone of high speed steel W6Mo5Cr4V2(M2)melt after laser fu-sion was found to be so fine as the ehill zone of a solidified ingot.When narrower chill zoneformed,the long columnar dendrites ...Microstructure of the deepest zone of high speed steel W6Mo5Cr4V2(M2)melt after laser fu-sion was found to be so fine as the ehill zone of a solidified ingot.When narrower chill zoneformed,the long columnar dendrites grow into the melt and then the fine equiaxed cellularstructure appears in upper melt region nearly surface.The substructure of cellular grains anddendrites was observed to consist of martensite and retained austenile,while the carbides asM_6C_■ Cr_7C_3 and MC distributed at their boundaries.It is believed that the highermicrohardness up to HV_(0.1)=865-960 of the laser fused structure of the alloy is due to the oc-currence of martensite.展开更多
基金Supported by a National Research Foundation(NRF)of Korea grant funded by the Korean government(Ministry of EducationScience+2 种基金and TechnologyMEST)Grant No.2013-005810
文摘AIM: To develop a Fok-I nested polymerase chain reaction(PCR)-restriction fragment length polymorphism analysis(PRA) method for the detection of hepatitis B virus X region(HBx) V5 M mutation.METHODS: Nested PCR was applied into DNAs from 198 chronic patients at 2 different stages [121 patients with hepatocellular carcinoma(HCC) and 77 carrier patients]. To identify V5 M mutants, digestion of nested PCR amplicons by the restriction enzyme Fok-I(GGA TGN9↓) was done. For size comparison, the enzymetreated products were analyzed by electrophoresis on 2.5% agarose gels, stained with ethidium bromide, and visualized on a UV transilluminator.RESULTS: The assay enabled the identification of 69 patients(sensitivity of 34.8%; 46 HCC patients and 23 carrier patients). Our data also showed that V5 M prevalence in HCC patients was significantly higher than in carrier patients(47.8%, 22/46 patients vs 0%, 0/23 patients, P < 0.001), suggesting that HBx Ag V5 M mutation may play a pivotal role in HCC generation in chronic patients with genotype C infections.CONCLUSION: The Fok-I nested PRA developed in this study is a reliable and cost-effective method to detect HBx Ag V5 M mutation in chronic patients with genotype C2 infection.
文摘Microstructure of the deepest zone of high speed steel W6Mo5Cr4V2(M2)melt after laser fu-sion was found to be so fine as the ehill zone of a solidified ingot.When narrower chill zoneformed,the long columnar dendrites grow into the melt and then the fine equiaxed cellularstructure appears in upper melt region nearly surface.The substructure of cellular grains anddendrites was observed to consist of martensite and retained austenile,while the carbides asM_6C_■ Cr_7C_3 and MC distributed at their boundaries.It is believed that the highermicrohardness up to HV_(0.1)=865-960 of the laser fused structure of the alloy is due to the oc-currence of martensite.
