The purpose of splenectomy was to improve hypersplenic thrombocytopenia and introduce IFN for clearance of the HCV virus. Forty-eight patients who underwent hepatectomy due to liver tumors were recruited as controls (control group 1). PB samples from 10 healthy adult volunteers (control group 2) and spleen tissues obtained by splenectomy from seven patients because of trauma (control group 3) were also used as controls. In addition, all patients were HIV negative. Patients received no medical treatment except splenectomy during the study period. All samples were studied after

obtaining EPZ-6438 supplier the appropriate institutional informed consent. We also obtained permission from the ethical review board. A total of 26 pieces from the resected liver specimens of patients with HCV-related liver cirrhosis and hypersplenism who underwent splenectomy were also examined for the immunohistochemical expression of CD4+ lymphocytes, CD8+ lymphocytes, FOXP3, granzyme B and TGF-β1 positive cells (Fig. 1). We classified liver specimens into five stages according to the degree of fibrosis as follows: F0, no fibrosis in the portal tract; F1, portal fibrosis without septa; F2, portal fibrosis with a few septa; F3, numerous septa without cirrhosis; and F4, cirrhosis. We

collected resected liver specimens from 10 cases PI3K activation each of F1, F2, F3 and F4 with HCV-related liver disease. We also collected specimens from eight cases of liver hemangioma of F0 with both negative hepatitis B surface antigen and HCV antibody. Follow-up liver biopsy sections find more were obtained from the same part of the liver if possible from seven of the 26 patients at various intervals after splenectomy

(Table 2). These sections were used for CD4 and CD8 immunostaining and Masson-trichrome staining for the morphometric evaluation of fibrotic areas. A total of 26 spleens with HCV-related liver cirrhosis and hypersplenism were examined for the immunohistochemical expression of CD4 positive lymphocytes, CD8 positive lymphocytes, FOXP3, granzyme B and TGF-β1 positive cells. We measured the same parameters in spleens from the seven control cases in control group 3 as a non-cirrhotic control (Fig. 1). Spleen and liver tissues were pathologically assessed by two pathologists (Y. N. and M. K.). Peripheral blood samples were serially collected from 26 patients with HCV-related liver cirrhosis and hypersplenism just before and 14 days, 1 month, 3 months, 6 months and 1 year after splenectomy. We examined the ratio of CD4+ T cells to all lymphocytes, CD8+ T cells to all lymphocytes, and the CD4+/CD8+ ratio in PB samples using flow cytometry. TGF-β1 levels in PB were also measured using enzyme-linked immunoassays in the sera just before and 14 days, 1 month, 3 months, 6 months and 1 year after splenectomy. Patients were excluded from the protocol if IFN or other therapeutics were introduced for the liver disease.