Objective To investigate the the quasispecies character of 3′ untranlated region(3′ UTR) in hepatitis C virus(HCV) by analysing the nucleotide sequence polymorphism and mutation features in 3′ UTR region.Methods Patients infected with genotype 1b HCV were identified by reverse transcription-nested polymerase chain reaction(RT-PCR) and restriction fragment length polymorphism(RFLP) assay.Fragments of the cDNA of 3′ UTR were amplified using semi-nested RT-PCR,and subjected to cloning.The 12-15 clones that contained HCV 3′UTR gene fragments amplified from each patients were sequenced.Results The full-length sequence of 1b genotype HCV 3′UTR in cDNA were obtained.The 3′UTR region consists of four elements: the 5′ region,the poly(U),poly(U/C) and 98-base region.The nucleotide sequence diversity ranged from 0.2%～2.1% and the mutation points were almost distributed in the the 5′region and poly(U/C).Conclusions The HCV has complex quasispecies character in 3′ UTR.

As to evaluating the medical compliance of patients with chronic hepatitis B (CHB) and the influence factors, information of 163 patients with CHB treated with lamivudine were collected by using the self-designed questionnaire, and the medical compliance and influencing factors of the patients were acquired by the research followed, and the prognosis and allocation were also observed. The results showed the medical compliance in 77.3% of patients with CHB was good, and in 22.7% patients was poor. After following-up for 2 years, the negative rates of HBV DNA and HBeAg shown in patients with good medical compliance were 78.6% and 46.0%, respectively,showing a significantly higher rate than in those with poor medical compliance, which having the rate of 37.8% and 21.6%, respectively (P<0.01). We suggested that there should be a correlation between the medical compliance of patients with CHB and the prognosis.

OBJECTIVETo obtain very end full-length cDNA of hepatitis C virus (HCV) 5' untranslated region (5' UTR), and analyse its primary and secondary structure.

METHODSBy reverse transcription-nested polymerase chain reaction (RT-PCR) and restriction fragment length polymorphism (RFLP), a patient infected with genotype 2a HCV was found. Total RNA isolated from the serum as template, the cDNA of 5' noncoding region was amplified using rapid amplification of cDNA ends methods (RACE), the fragments were recombined by A-T clone strategy, the recombinants were confirmed by RFLP and PCR then sequenced. Secondary structures were analysed by RNA draw.

RESULTSVery end full-length cDNA of 2a genotype HCV 5' UTR was obtained by RACE. In five clones obtained, three contained full-length 5' UTR cDNA, and A21G, G170A, T222C, T247C, C339T substitutions were found compared with HC-J6. he homologies with HCV-1,HC-J6,HC-C2, HC-J8 were 93.6%-94.4%, 92.1%-93.0%, 98.8%-99.7%, 96.2%-96.5%, respectively; however, the substitutions did not alter the secondary structure. Two out of five clones were deleted to have 53 and 144 bases at 5' terminus of HCV 5' UTR, respectively.

CONCLUSIONSRACE is rapid and effective, works well to obtain very end of virus genome. With that, Authors obtained full-length cDNA of genotype 2a of HCV 5' UTR. There are genes deleted at 5' terminus circulated in hepatitis C patients.

5' Untranslated Regions ; genetics ; Base Sequence ; DNA, Complementary ; genetics ; DNA, Viral ; genetics ; Hepacivirus ; classification ; genetics ; isolation & purification ; Hepatitis C ; virology ; Humans ; Molecular Sequence Data ; Nucleic Acid Amplification Techniques ; methods ; Polymorphism, Restriction Fragment Length ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA

3' Untranslated Regions ; genetics ; Base Sequence ; China ; DNA, Complementary ; chemistry ; genetics ; Genetic Variation ; Hepacivirus ; genetics ; Humans ; Molecular Sequence Data ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Alignment ; Sequence Analysis, DNA ; Sequence Homology, Nucleic Acid