Total RNA was isolated from human plasma and RNA concentrations were determined using a Nanodrop 1000 spectrophotometer. cDNA was generated from the RNA and microRNA assays were performed by real-time PCR using the specific primers for human miR-33a, miR-103, and miR-155. Expression levels of cel-miR-39, RNU6B, and mir-16 were used as the endogenous controls for assays of miR-33, miR-103, and miR-155, respectively. To quantify the relative expression level of each miRNA, threshold cycle (Ct) values were normalized against endogenous reference (ΔCt = Ct (target miR)−Ct (endogenous control)). Expression levels of human plasma miRNAs were calculated by the 2−ΔCt method. Each microRNA was assayed in triplicates.