Abstract: Objective To explore the level of serum 25（OH）D₃ in male undergoing advanced health checkup from 2014 to 2018 and investigate its related influencing factors. Methods Totally 37 853 males hospitalized in the International Medical Center for health checkup were recruited in our study from 2014 to 2018.The trends of serum 25（OH）D₃ level in different years and age groups were analyzed,and the serum 25（OH）D₃ levels were compared between deficiency group,lack group and sufficient group in each year. Results The serum concentration of 25（OH）D₃ in these population demonstrated an increasing trend over time,with （16.61±7.14） ng/mlin 2014,（18.40±7.44） ng/ml in 2015,（20.45±7.99） ng/ml in 2016,（20.20±6.37） ng/ml in 2017,（21.03±5.60） ng/ml in 2018 （P for trend=0.000）.Most of the subjects were in vitamin D deficiency status.The rate of vitamin D deficiency was 73.90% in 2014,64.50% in 2015,57.20% in 2016,55.44% in 2017,48.70% in 2018,showing a decreasing trend （P for trend=0.000）.Vitamin D level increased with aging,with the highest level in 79-80 age group in 2014,2015,2016,2017,respectively. Conclusion The level of 25（OH）D₃ in male subjects is generally low,but it increases year by year.