| AIM: Conventionally, chronic infection with HBV is defined via the lasting presence of HBs in the serum for at least 6 months. Regardless if detecting chronic HBV infection in clinical or not, and if causing progressive liver disease or not, the chronic HBV infection appears to increase the risk for the development of HCC. Although vaccination programs in several countries have been successful, for those already infected, it is important to develop discriminative screening procedures as a basis for targeted therapy of critical infection stages. The present study introduces circulating viral transcripts as a potential parameter.The four overlapping ORFs on viral DNA, i.e., PreC/C, polymerase, PreS1/PreS2/S, and X, are transcribed into genomic and subgenomic RNA molecules. The translation product of the smallest viral transcript, HBx, is regarded as a hepatocarcinogenic factor.Properties assigned to HBx include the stimulation of transcription and signal transduction, interference with DNA repair, induction of apoptosis, and malignant transformation both in vitro and in vivo. During replication of HBV, all viral transcripts mature at a unique poly(A) signal downstream of the HBx ORF, namely a UAUAAA motif at position 1789. Collectively, these transcripts are referred to as full-length tanscripts in this body. Other transcripts, designated as truncated, use for maturation a cryptic poly(A) signal, a CAUAAA motif within the HBx ORF (position 1661). TrRNA was hidden in the HBx ORFs, and only when the mature signal of fRNA was inactivation, should trRNA have activition. The method to detect fRNA and trRNA of HBV transcripts in liver tissue had been established by Schutz et al in 1996. Su and Zhang et al established the method to detect the two kinds of viral transcripts in serum later. So these two methods were used in our experiment. Serum of 145 patients who have liver cancer, hepatocirrhosis, and other liver disease with HBsAg positive and negative were collected to extract nucleic acid, and to detect HBx DNA, fRNA and trRNA by PCR and RT-PCR from nucleic acid. Tissues of 49 patients who have liver surgical operation were collected to extract nucleic acid, and HBx DNA and RNA were detected. We analyzed the results in serum and tissue of all patients and contrasted the resluts of serum and tissue in the same patient for the first time which have not been reported now, and to explore the relationship between the HBV transcript and end-stage liver disease.Methods: Experiment one: 145 patients who is in hospital in 2002-12 / 2003-08 were selected from general surgery of Xi-jing and Tang-du hospital of our university. They are 102 males and 43 females, 8~73y youngest and oldest (average 46.68y), 67 cases of liver cancerpatients, 46 cases of hepatocirrhosis patients, 19 cases of other liver disease patients and 13 cases of non-liver disease patients, also 70 cases of HBsAg positive patients which included 20 HBsAg positive and HBeAg positive patients, 75 cases of HBsAg negative patients which included 32 non-HBV immunologic mark patients.Venous blood of 145 patients were drawed in the moring and inject to the sterile, anticoagulantal and desiccated tube which would be centrifuged at 4C, 3000g, for 25 minutes at once. The serum were separated from upper suspension, transferred to a new eppendorf tube, and stored at -70C .Nucleic acid in serum was extracted by using high pure viral nucleic acid kit and its final volume was 50ul. Using nucleic acid as template and txs+/txas- as primer, HBx DNA was amplified by PCR. HBV transcripts fRNA and trRNA were amplified by two-times PCR, which used one-step method Titan one tube RT-PCR system and has the same template as HBx DNA amplification, and the primer is txs+/ txs2+/txass-. Their products of PCR and RT-PCR were displayed by agarose electrophoresis and to be analyzed.Experiment two: 49 patients of 145 which has surgical operation of liver were selected from 145 patients. They are 32 males and 17 females, 8~68 youngest and oldest, 16 cases of liver...
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