| Objectives: (1) To evaluate the distribution of histopathologic parameters and biological markers and the association between them in the progression from chronic hepatitis C to cirrhosis and hepatocellular carcinoma. To compare predictive superiority among the parameters and search for reliable predictors of outcome in chronic hepatitis C. (2) To assess the relationship of host factors, viral factors and biochemical indices and the efficacy of interferon (IFN) theropy. (3) To clarify the expression of CD34, CD31, vWF,α-SMA, Ki-67 and Bcl-2 and clinicopathologic significance in HCC. To compare the different expression of these markers between HCV-associated HCC and HBV-associated HCC. (4) To compare the expression of CD34, CD31 and Ki-67 among normal liver, chronic hepatitis, cirrhosis, HCC and liver metastatic adenocarcinoma and search for valuable markers to distinguish different diseases in liver. (5) According to the above-mentioned studies, to clarify the mechanism of progression from chronic hepatitis C to HCC. (6) To analyze the characteristic of ultrasound, especially three-dimensional color power angiography (3D-CPA) for primary liver cancer and its relation to clinicopathologic significance.Methods: (1) Subjects 73 patients with chronic hepatitis C who had undergone ultrasonography-guided needle liver biopsy at Suwa Red Cross hospital, Japan, from March 1993, to Norvember 1999, were involved in this study. The patients inclued 37 men and 36 women, with ages from 18 to 72 years (52.4±11.3 years). All patients were followed up at least 3 years from biopsy and screened, 9 patients developeddecompensated cirrhosis and 4 patients developed HCC. To compare and analyze all patiens divided into obvious progression group and no obvious progression group. Of 73, 60 patients including 31 men and 29 women, with ages from 18 to 72 years (52.3±11.7 years), were treated by IFN, with the conditions of first IFN therapy and no treatment in 6 months. The patients with HCC divided into two groups; One was composed of 29 patients who underwent partial hepatectomy in the Fourth Hospital of Hebei Medical University from Septemper 2000 to Norvember 2001, including 24 men and 5 women with ages from 31 to 68 years (51.6±9.3 years), the other was 7 patients who underwent hepatectomy in Suwa Red Cross Hospital, Japan, 24 men and 5 women with ages from 52 to 73 years (60.1±7.6 years). All patients had no treatment before operation. There were 10 cases of normal liver and 7 cases of cirrhosis. 7 patients with liver metastatic adenocarcinoma originating from colon and breast were also involved in this study. Ultrasound detections were performed in 248 patients with HCC, of 248 patients, 58 patients were examined by 3D-CPA; contrastive study of 3D-CPA with pathologic outcome was performed in 30 patients of HCC. (2) Histopathology Liver biopsy specimens of chronic hepatitis C were routinely fixed in 10% buffered formalin, embedded in paraffin. These sections were stained with hematoxylin and eosin (HE), Azan-Marroy, and periodic acid Schiff reaction (PAS). Each specimen consisted of at least 5 portal tracts and was evaluated for portal inflammation, piecemeal necrosis and bridging necrosis, intralobular degeneration and necrosis of hepatocytes, fibrosis and numerically scored according to the histology activity index (HIA) scoring system proposed by Knodell et al. (3) Immunohistochemistry All specimens were fixed in 10% buffered formalin and embedded in paraffin. The deparaffinized sections were treated in 0.3% hydrogen peroxide in methanol for 30 minutes to eliminate endogenous peroxidase activity. For CD31, vWF,α-SMA, Ki-67, Bcl-2, retrieval of antigenecity was performed by microwave, slides were placed in 10 mmol/L citrate buffer (pH6.0), boiled for 15minutes in a microwave oven, cooled for 15 minutes, and rinsed in distilled water. For CD34, retrieval of antigenecity was performed by trypsin, sections were put into 0.2% trypsin buffer at 37℃, and then at 64℃ for 45 minutes. Primary antibodies...
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