17 HCV-RNA was determined using the COBAS TaqMan HCV test (Roche Diagnostics, Basel, Switzerland). The serum samples stored at −80°C before IFN therapy were used. The linear dynamic range of the assay was 1.2-7.8 log IU/mL, and the undetectable samples were defined selleckchem as negative. A sustained virological response (SVR) was defined as clearance of HCV-RNA using the COBAS TaqMan HCV test 6 months after

the cessation of IFN therapy. Status of liver was mainly determined on the basis of peritoneoscopy and/or liver biopsy. Liver biopsy specimens were obtained using a modified Vim Silverman needle with an internal diameter of 2 mm (Tohoku University style; Kakinuma Factory, Tokyo, Japan), fixed in 10% formalin, and stained with hematoxylin and eosin, Masson’s trichrome, silver impregnation, and periodic acid-Schiff after diastase digestion. The size of specimens for examination was more than six portal areas.18 The observation Smad inhibitor starting point was 6 months after the termination of IFN therapy. After that, patients were

followed up at least twice a year in our hospital. Physical examination and biochemical tests were conducted at each examination together with a regular checkup. In addition, annual examinations during follow-up were undertaken. When a patient had complaints during follow-up, the physician check details in charge performed additional examinations based on symptoms.

Four hundred eighteen patients were lost to follow-up. The final date of follow-up in 418 patients with loss of follow-up was regarded as the last consulting day. In addition, 881 patients were retreated with IFN. The final date of follow-up in 881 patients re-treated with IFN were regarded as the time of the initiation of IFN retreatment. Thus, 418 patients with loss of follow-up and 881 patients retreated with IFN were counted censored data in statistical analysis.19 The mean follow-up period was 6.8 (SD 4.3) years in 418 patients with loss of follow-up and 7.5 (SD 4.8) years in 881 patients retreated with IFN. Censored patients were counted in the analysis. Clinical differences among three groups of patients with HCC with malignancies other than HCC without events were evaluated using the Kruskal-Wallis test. The cumulative development rates of malignancies were calculated using the Kaplan-Meier technique, and differences in the curves were tested using the log-rank test.20,21 Independent risk factors associated with malignancies were studied using the stepwise Cox regression analysis.