Expression of genes relative to GAPDH was calculated based on the threshold cycle (Ct) as 2?��(��Ct), where promotion info ��Ct=Ct,GENE?Ct,GAPDH and �� (��Ct)=��Ct,N?��Ct,T (N, matched normal tissue cDNA; T, tumor tissue cDNA; NN, normal tissue cDNA from patients without cancer). Primer sequences are shown in Table S3. Immunohistochemistry Tissue microarrays were performed with sections (5 ��m) of colon cancer tissues, adjacent tissues 1.5 cm away from tumor, and non-malignant normal colon tissues which were purchased from US Biomax, Inc. (Rockville, MD). The tissues were deparaffinized and incubated with anti-OSMR rabbit polyclonal antibody (1100 dilution) (Santa Cruz Biotechnology, Santa Cruz, CA) at 4��C overnight. They were then incubated in broad spectrum secondary antibody purchased from DAKO (Carpinteria, CA) for 30 min.
After washing the slides in PBS, tissue sections were stained with freshly prepared DAB chromogen solution (DAKO). We treated tissues with streptavidin and biotin (Invitrogen) for 20 min each to block endogenous biotin levels. Sections were counterstained in Mayer’s Hematoxyline. Luciferase reporter assay The pGL3-OSMR-
Hepatitis B virus (HBV) affects over 350 million people worldwide while countries in Asia and Africa account for over 70% of chronic HBV infection, with prevalence up to 15%�C20% [1], [2]. In China, it was estimated that at least 10% of the general population are chronically infected with HBV, which becomes the most common cause of liver diseases [3].
Though the efficacy of antiviral therapy in chronic hepatitis B (CHB) has been greatly improved in the last decades after discovery of interferon and nucleoside analogues, lack of response still remains common [4]. It is well recognized that uncontrolled virus replication can cause liver damage and predispose those nonresponders into liver diseases of advanced stage. Therefore, unraveling factors associated with treatment failure in CHB patients is of clinical importance. Nonalcoholic fatty liver disease (NAFLD) is defined as a common clinico- pathologic condition characterized by lipid deposition with/without inflammation in hepatocytes and comprises a wide spectrum of liver damage, including simple steatosis, nonalcoholic steatohepatitis (NASH) and fibrosis [5]. With social development and lifestyle change, NAFLD has now become a major cause of liver related morbidity and mortality, with the incidence of around 20% worldwide [6] and 15% in China [7].
Therefore, the coexistence of HBV infection and NAFLD becomes a novel characteristic of liver disease in China. However, their bilateral influence in both disease development and therapeutic response has been rarely reported. Hepatic steatosis has long been considered as a common hepatocellular change in both simple Carfilzomib steatosis and NASH.