Objective To investigate and evaluate the method of PCR amplification blocking associated with fluorescent probe for the detection of pre-C region of HBV G1896A gene mutation.Methods The primers were designed based on the mutation of HBV DNA 1896 locus.The 3′end of primers was at 1896 site,and it was complemented with the base sequence of mutation template of 1896 site.The mismatching bases were separately introduced into the second and the third base of the primer by inverse counting from the 3′end for increasing the specificity of reaction.Results The PCR amplification for wild plasmid with the mutant primer showed an effectively blocking,but not showed blocking for the mutant plasmid (G1896A).The sensitivity of detection for the mutant plasmid was 5?103 copies/ml.Ninety-five cases of HBV-positive serum was selected randomly and amplified with the mutant primer,and 8 cases were positive HBV G1896A gene mutant(mutant rate of 8.4%).Conclusion The amplification blocking associated with fluorescent probe for the detection of HBV G1896A gene mutation is a effective,convenient method for the detection of clinical samples.

PURPOSE: This study was undertaken to show the clinical results of combined radiotherapy and hyperthermia in primary hepatoma. MATERIALS AND METHODS: Between December 1989 and March 1993, 50 patients with hepatomas were treated by combined radiotherapy and hyperthermia. Among them, we analyzed retrospectively 33 patients who received the complete course of treatment. The ages of the patients ranged from 36 to 75(mean age: 55.5 years). Twenty-six patients (78.8%) were men, and 7 (21.2%) were women. According to Child's classification, nine patients (27.3%) were A group, 9 (27.3%) were B group, 15 (45.4%) were C group. Radiation therapy was done by a 6 MV and 15 MV linear accelerator. Patients were treated with daily fractions of 150-180 cGy to doses of 2550 cGy - 4950 cGy (median: 3000 cGy). Local hyperthermia was done by 8 MHz RF capacitive heating device (Cancermia. Green Cross Co., Korea), 50-60 min/session, 1-2 sessions/wk, and 8.5 sessions (median number)/patients. We analyzed the prognostic factors including age, sex, tumor type, Child's classification, a-fetoprotein, liver cirrhosis, ascites, portal vein invasion, esophageal varix, number of hyperthermia, chemotherapy, total bilirubin level, Karnofsky performance status. RESULTS: The overall 1-year survival was 24.2%, with a mean survival of 10 months. Of 33 patients, tumor regression (PR+MR) was seen in 30.4%, no response was seen in 52.2%, 17.4% patient was progressed. In patients who had tumor regression, the overall 1-year survival was 42.1% with a mean survival of 14 months. Factors influencing the survival were sex (p=0.05), tumor type (p=0.0248), Child's classification (p=0.0001), liver cirrhosis (p=0.0108), ascites (p=0.0009), and Karnofsky performance status (p=0.0028). Complications developed in 28 patients, including 18 hot pain, 5 fat necrosis, 3 transient fever, 2 nausea and vomiting. CONCLUSION: In this study, the results suggests that combined radiotherapy and hyperthermia may improve the survival rate of hepatoma.
Ascites ; Bilirubin ; Carcinoma, Hepatocellular* ; Classification ; Drug Therapy ; Esophageal and Gastric Varices ; Fat Necrosis ; Female ; Fever* ; Heating ; Hot Temperature ; Humans ; Hyperthermia, Induced ; Karnofsky Performance Status ; Liver Cirrhosis ; Male ; Nausea ; Particle Accelerators ; Portal Vein ; Radiotherapy* ; Retrospective Studies ; Survival Rate ; Vomiting