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Objective To study the relationship between semen quality and the methylation level of maternally expressed gene 3 (MEG3) in sperm. METHODS We collected the general demographic data, semen samples and results of clinical semen analysis from 403 married men undergoing pre-conception examinations in March and April 2015 and March and April 2016. Using pyrosequencing, we quantitatively detected the methylation level at 8 CpG sites in the differentially methylated region of MEG3, and subjected the data obtained to variance analysis, Pearson correlation analysis