The production of each miRNA was quantified with the StepOne Real-Time PCR System (Applied Biosystems). All reactions were carried out in triplicate. The mean value of the threshold cycle (Ct), the intersection between the amplification curve and the threshold line, was normalized using the value of RNU48, a small RNA serving as endogenous control. In addition, a single sample was used as the calibrator sample to correct the values. Then, 2-ΔΔCt values find more were calculated as relative values.[19] Using a comparative Ct method, relative

expressions of each miRNA were compared between healthy controls and patients with each disease. Parameters related to clinical presentation for miRNA and PBC included ALT, ALP, GGT, total bilirubin (TB), IgG, IgM, and AMA-M2 at the time of miRNA sampling. In addition, histopathological assessment was performed according to the new histologic and grading system for PBC.[20] Briefly, scores for fibrosis and bile duct loss were combined for staging: stage 1, total score of 0; stage 2, score of 1–2; stage 3, score of 3–4; and stage 4, score of 5–6. Cholangitis activity (CA) and hepatitis activity (HA) were graded as CA0-3 and HA0-3, respectively. Response to PBC treatment was defined by a decrease in ALP of more than 40% of the baseline

value or normal level within 1 year of treatment with ursodeoxycholic acid (UDCA) at a maximum dose of 900 mg/day (n = 7), according to the Barcelona criteria.[21] Administration of Bezafibrate within selleck chemicals llc one year after the start of UDCA therapy in some patients was decided on the basis of response to monotherapy with UDCA. Blood samples from PBC patients were obtained during treatment with UDCA and/or bezafibrate. Similarly, blood samples from patients with AIH, PBC-AIH selleckchem overlap and SLE were obtained during treatment with prednisolone (5–10 mg/day) and/or UDCA (600 mg/day). The baseline characteristics of PBC and other patients at the time of miRNA sampling

are summarized in Table 1. Data were expressed as mean ± standard deviation (SD). Statistical analysis was performed using Student’s t-test, Pearson’s correlation coefficient and differences were considered statistically significant when the P-value was less than 0.05 in the two-sided test. As shown in Figure 1, there were significant differences in the expression of some miRNAs between healthy controls and patients with autoimmune liver diseases. In PBC, expressions of miR-155 and miR-146a were significantly increased compared to those in healthy controls. Similarly, increased miR-155 expression and decreased miR-26a expression were observed in AIH, and significantly increased expression of miR-155 was observed in PBC-AIH overlap syndrome. In SLE, expressions of miR-155 and miR-16 were significantly increased compared to those in healthy controls.