Briefly, 1 × Probes Master, 200 nM of each primer,

100 nM Universal ProbeLibrary probe, and 2 μl diluted cDNA template were added to each reaction in a total volume of 20 μl. The protocol consisted of an initial denaturation step at 95°C for 10 min, followed by 40 cycles of amplification and quantification at 95°C for 15 s, 60°C for 10 s, and 72°C for 10 s, and was finally cooled at 40°C. The transcript amounts were estimated from the respective standard curves and normalized to the GAPDH transcript amount determined in corresponding samples. Reactions were run in duplicate. Statistical analysis Results are presented as mean ± SEM. Differences of mean expression levels between groups were compared with the student t-test or Welch’s t-test. Associations were assessed by Pearson’s correlation coefficient test or

Spearman’s GSK3235025 solubility dmso rank-correlation coefficient test, and expressed by the corresponding correlation coefficient (rs). Curves of native liver survival were calculated using Kaplan-Meier methodology and log rank test was used to compare survival rates. P values < 0.05 were considered significant. References 1. Hartley JL, Davenport M, Kelly DA: Selleckchem mTOR inhibitor biliary atresia. Lancet 2009, 374:1704–1713.PubMedCrossRef 2. Schweizer P: Treatment of extrahepatic biliary atresia: results and long-term prognosis after hepatic portoenterostomy. HMPL-504 ic50 Pediatr Surg International 1986, 1:30–36. 3. Ohi R: Biliary atresia: a surgical perspective. Clin Liver Dis 2000, 4:779–804.PubMedCrossRef 4. Sokol RJ, Mack C, Narkewicz MR, Karrer FM: Pathogenesis and outcome of biliary atresia: current concepts. J Pediatr Gastroenterol Nutr 2003, 37:4–21.PubMedCrossRef 5. Shneider BL, Brown MB, Haber B, Whitington PF, Schwarz K, Squires R, Bezerra J, Shepherd R, Rosenthal P, Hoofnagle JH, Sokol RJ, Biliary Atresia Research Consortium: A multicenter study of the outcome

of biliary atresia in the United States, 1997 to 2000. J Pediatr 2006, 148:467–474.PubMedCrossRef 6. Davenport M, Howard ER: Macroscopic appearance at portoenterostomy-a prognostic variable in biliary atresia. J Pediatr Surg 1996, 31:1387–1390.PubMedCrossRef 7. Davenport M, Caponcelli Rapamycin ic50 E, Livesey E, Hadzic N, Howard E: Surgical outcome in biliary atresia: etiology affects the influence of age at surgery. Ann Surg 2008, 247:694–698.PubMedCrossRef 8. Gautier M, Jehan P, Odievre M: Histologic study of biliary fibrous remnants in 48 cases of extrahepatic biliary atresia: correlation with postoperative bile flow restoration. J Pediatr 1976, 9:704–709. 9. Hitch DC, Shikes RH, Lilly JR: Determinants of survival after Kasai’s operation for biliary atresia using actuarial analysis. J Pediatr Surg 1979, 14:310–314.PubMedCrossRef 10.