RESUMEN
Background: Epigenetic DNA methylation is an essential mechanism controlling gene expression and cellular function. Existing analyses with conventional assays have generated significant insights into static states of DNA methylation, but were unable to visualize the dynamics of epigenetic regulation. Aim: We utilized a genomic DNA methylation reporter (GMR) system to track changes in DNA methylation during cardiac differentiation. Methods and Results: The promoter region of Cdk1 (Cyclin-dependent kinase 1) or Sox2 (SRY-Box Transcription Factor 2) gene was cloned upstream of the small nuclear ribonucleoprotein polypeptide N (Snrpn) minimal promoter followed by a fluorescent reporter gene. Mouse induced pluripotent stem cells (iPSCs) carrying Sox2 GMR rapidly lost fluorescent reporter signal upon the induction of differentiation. Cdk1 GMR reporter signal was strong in undifferentiated iPSCs, and gradually decreased during directed cardiomyocyte (CM) differentiation. RT-qPCR and pyrosequencing demonstrated that the reduction of Sox2 and Cdk1 was regulated by hypermethylation of their CpG regions during cardiac differentiation. The present study demonstrated the dynamic DNA methylation along the course of cell cycle withdrawal during CM differentiation. Conclusion: The GMR reporter system can be a useful tool to monitor real-time epigenetic DNA modification at single-cell resolution.
RESUMEN
Background: Global warming, lifestyle, or working in a high temperature environment leads to have increased health risk factors. This meta-analysis was conducted to determine the impact of high ambient temperature on male reproductive function. Methods: Scientific articles were screened in the database including MEDLINE, EMBASE, National center for biotechnology information (NCBI) or Web of Science with relating keywords. Impact data of high ambient temperature on semen parameters were extracted and analyzed by STATA software according to the Random Effects Model. The high ambient temperature exposure group and Non-exposure group were compared using the standard mean difference (SMD). Publications were evaluated for publication bias by Egger test. Results: Nine articles were finally selected from databases examining the effect of high ambient temperature on male reproductive health of 356 men from Iran, Italy, Thailand, China, Egypt. High ambient temperature showed a significant decrease in the seminal parameters, semen volume during each ejaculation (SMD = -0.74; 95% CI -1.11, -0.36), sperm concentration (SMD = -1.07; 95% CI -1.42, -0.72), total sperm count (SMD = -1.52; 95% CI -2.96, -0.08), sperm motility (SMD = -1.93; 95% CI -2.83, -1.04), sperm progressive motility (SMD = -1.65; 95% CI -2.39, -0.91) and normal morphology (SMD = -2.41; 95% CI -3.30, -1.52). Conclusion: High ambient temperature negatively affects sperm quality, including decreased semen volume, sperm count, sperm concentration, motility and normal morphology. This might lead to protective strategies to avoid the adverse effects of high ambient temperature on male fertility.