Abstract: Objective To explore the relationship between 25 hydroxyvitamin D［25(OH)D］deficiency and seasons, dyslipidemia, overweight or obesity. Methods A total of 2 677 healthy subjects with physical examination from September 2017 to August 2019 were collected and divided into three groups according to serum level of 25(OH)D. The recruiters with level of 25(OH)D>30 ng/mL was included in normal group, those with level of 25(OH)D>20~30 ng/mL was selected as 25(OH)D insufficiency group, and those with level of 25(OH)D ≤ 20 ng/mL was selected as 25(OH)D deficiency group. The body mass index(BMI), serum lipid index levels among three groups, the distribution of 25(OH)D levels and nutritional status in four seasons were compared. Results 25(OH)D detection results showed that the ratios of patients with 25(OH)D deficiency, insufficiency and normal were 66.23%(1 773/2 677), 25.03%(670/2 677), 8.74%(234/2 677), respectively. The highest 25(OH)D level was 19.92 ng/mL in the summer group, followed by 17.54 ng/mL in the autumn group and 15.95 ng/mL in the spring group, and the lowest in the winter group(13.61 ng/mL). The - between-group difference was statistically significant(P<0.01). The highest 25(OH)D deficiency rate was 78.61% in the winter group and its lowest rate was 51.34% in the summer group. The highest 25(OH)D normal rate was 13.55% in the summer group, which was significantly higher than that in the spring, autumn and winter groups(P<0.01). BMI and triglycerides(TG)increased in the 25(OH)D normal, deficiency, insufficiency groups, while high-density lipoprotein cholesterol(HDL-C), and apolipoprotein A1(ApoA1)decreased, with statistical significant difference(P<0.05 or P<0.01). Logistic regression analysis showed that after adjusting age and gender, risks of abnormity in TG(OR=2.273, 95%CI: 1.636~3.159, P<0.001), HDL-C(OR=2.817, 95%CI: 1.783~4.464, P<0.001), and overweight/obesity(OR=1.892, 95%CI: 1.394~2.569, P<0.005)were all increased compared with those with normal 25(OH)D level. Compared with those with normal 25(OH)D level, risks of abnormity in TG(OR=2.100, 95%CI: 1.483~2.974, P<0.001), HDL-C(OR=2.183, 95%CI: 1.353~3.534, P<0.001), and overweight/obesity(OR=1.603, 95%CI: 1.155~2.224, P<0.001)were all increased. Conclusion Deficiency of 25(OH)D is affected by the seasonal changes, and is directly related to the abnormal blood lipid and overweight/obesity.