Statistical and Bioinformatic Analy

Statistical and Bioinformatic AnalysesStatistical analysis was performed using PASW v 18, IBM SPSS Statistics 21 (SPSS Inc, Chicago, Illinois) or GraphPad Prism v 5 (GraphPad Software Inc, San Diego, California).Statistical comparisons of clinical data between cases and controls were performed using ANOVA, Tukey post hoc tests, Mann-Whitney test, and Kruskal-Wallis tests, as appropriate. The threshold of statistical significance was set at a Pvalue of .05.Bioinformatic analysis was also performed. All samples in the initial exploratory study were analyzed for miRNA expression using human miRNA microarray 8  15 K release 14.0 (029297) arrays. The data from the microarrays were normalized using variance stabilizing normalization.16 The data were filtered to remove the control probes and those that did not pass the quality filters (ie, those that were undetectable or were not expressed higher than the negative controls in greater than 75% of the samples). After filtering, 259 miRNAs out of 866 were retained for differential expression analysis. All data processing and differential expression analysis was carried out in R using the AgiMicroRNA17 and LIMMA18 packages from Bioconductor 19 P values were corrected for multiple testing using the BenjaminiHochberg method.20Potential molecular targets and signalling pathways of hsa-miR-374a were identified using miRecords. Additional enrichment analyses were carried out using WebGestalt