Project description:Objective:This study aims to discover circulating exosomal miRNAs as potential noninvasive biomarkers early detection of fetus with ventricular septal defect (VSD). Method:A total of 182 pregnant women including 91 VSD cases and 91 matched controls were included in this study. Exosomes were isolated and next-generation sequencing was used to obtain a profile of dysregulated exosomal miRNA. The differential abundance was verified by quantitative real-time PCR (q RT-PCR). . Receiver operating characteristic (ROC) curves were conducted to evaluate the diagnostic accucary Results: 77 serum exosomal miRNA were uncovered to be differentially expressed in the VSD group as compared to the control group. Among these, five down-regulated exosomal miRNA were validated by qRT-PCR. hsa-miR-146a-5p was identified to be capable of distinguishing VSDs from controls (area under the ROC (AUC): 0.997; p < 2.2e-16). Conclusion: circulating exosomal miRNA, in particular, hsa-miR-146a-5pmay be a predictive biomarker for non-invasive prenatal diagnosis of fetal VSDs.
Project description:To identify the molecular markers of early pregnancy in pigs, we compared global gene expression profiles of the maternal peripheral blood in pregnant sows with the non-pregnant sows. Peripheral blood sample was collected at 14 days after insemination from the submandibular vein of pregnant and non-pregnant sows respectively, and total RNA was isolated, purified and sent for microarray analysis. This study identified 127 up-regulated and 56 down-regulated genes (FC >= 1.5 and P < 0.05) in peripheral blood from pregnant sows versus non-pregnant sows. Of the differently expressed genes, nine (including LPAR3, RXFP4, GALP, CBR1, CBR2, GPX6, USP18, LHB and NR5A1) were found to exert function related to early pregnancy processes. Seven differentially expressed genes (CHGB, USP18, VWF, LPAR3, NR5A1, PPARD, BIN1) were selected to perform qRT-PCR in the same RNA samples.The expression profiles of these genes detected by qRT-PCR were consistent with those by microarray, which confirmed the reliability of our microarray data.
Project description:We report the application of miRNA next generation sequencing (NGS) for the analysis of impact of processing on miRNA in human breast milk, donated by 3 volunteers. MiRNA content of total and exosomal fraction was compared between unprocessed milk and sample subjected to either Holder (thermal) pasteurization (HoP) or elevated pressure processing (HPP). NGS reads were mapped to miRBase in order to obtain miRNA counts. Then, we analyzed differences in the miRNA abundance and function between raw and processed material. It was observed that both processing methods reduce number of miRNA reads and HoP is significantly more detrimental to miRNA than HPP.
Project description:To investigate the exosomal miRNA changes under LPS treatment in RAW 264.7 cells, 2 μg/mL LPS were added into complete medium to incubate RAW 264.7 cells. And then The exosomes were isolated and tested the exosomal miRNAs change using microarray.