RNA sequencing identified specific circulating miRNA biomarkers for early detection of diabetes retinopathy.

Diabetic retinopathy (DR) is the leading cause of blindness in patients with diabetes. However, biomarkers for early detection of DR are still lacking. MicroRNAs (miRNAs) regulate multiple biological functions and are often deregulated in DR. We aimed to investigate whether circulating miRNAs can be used as biomarkers of early-stage DR. We used RNA-seq and qRT-PCR to identify differential serum miRNAs in patients with type 2 diabetes mellitus with DR (T2DM-DR), T2DM without DR (T2DM-no-DR), and healthy controls. We validated differential circulating miRNAs in two phases using qRT-PCR assays. RNA-seq analysis identified 7 differential circulating miRNAs between T2DM-DR and T2DM-no-DR and 47 differential miRNAs between T2DM-DR and healthy subjects. Two-stage analysis verified that a profile of five serum miRNAs (hsa-let-7a-5p, hsa-miR-novel-chr5_15976, hsa-miR-28-3p, has-miR-151a-5p, has-miR-148a-3p) was significantly associated with T2DM-DR. Receiver-operator-characteristic analyses showed that a panel of three miRNAs (hsa-let-7a-5p, hsa-miR-novel-chr5_15976, and hsa-miR-28-3p) presented 0.92 sensitivity and 0.94 specificity for distinguishing T2DM-DR from T2DM-no-DR, and 0.93 sensitivity and 0.86 specificity for differentiating early-stage T2DM-DR (NPDR) from late-stage DR (PDR). Lentivirus-mediated overexpression of hsa-let-7a-5p in human retinal microvascular endothelial cells (HRMECs) significantly promoted proliferation rates of HRMECs. In conclusion, the three-miRNA signature from serum may serve as a noninvasive diagnostic biomarker for DR. Furthermore, we showed that DR-associated miRNAs may be involved in the pathogenesis of DR, at least in part, through modifying proliferation of HRMECs.

Optimal timing of oocyte maturation and its relationship with the spindle assembly and developmental competence of in vitro matured human oocytes.

OBJECTIVE: To develop and test a simple, feasible approach to improve spindle assembly and developmental competence of human in vitro matured oocytes by parthenogenetic activation in maturation medium. DESIGN: Prospective, randomized study. SETTING: University research laboratory-based assisted reproductive technology laboratory. PATIENT(S): Four hundred thirty-two patients with male factor infertility undergoing intracytoplasmic sperm injection (ICSI) cycles. INTERVENTION(S): In vivo failed-to-mature oocytes from the ICSI cycles were divided into four groups according to differing duration after the extrusion of first polar body (0-1, 2-3, 4-5, and 8-9 hours). Oocytes spindles in each group were immunostained for α-tubulin and chromosomes, and observed by confocal microscopy. MAIN OUTCOME MEASURE(S): Rate from pronuclear stage to blastocyst, embryo grading at the eight-cell and blastocyst stages, and spindle assembly. RESULT(S): There was a statistically significantly higher rate of development at the eight-cell and blastocyst stages in the 2- to 3-hour and 4- to 5-hour groups. The grading results at the eight-cell and blastocyst stages also showed that the proportion of embryos of high quality was similar among the 2- to 3-hour and 4- to 5-hour groups, and in these groups it was statistically significantly higher than the 0- to 1-hour and 8- to 9-hour groups. The results of immunofluorescence demonstrated that there was a statistically significantly higher rate of normal spindle assembly in the 2- to 3-hour and 4- to 5-hour groups. CONCLUSION(S): Optimal timing of maturation benefits the development of competence of in vitro matured oocytes by promoting normal spindle assembly.