Objective · To investigate association between serum 25-hydroxyvitamin D3 [25(OH)D3] level and diabetic peripheral neuropathy (DPN) in elderly patients with type 2 diabetes mellitus and explore its value in clinical practice for early screening and prevention of DPN. Methods · A total of 188 elderly patients with type 2 diabetes mellitus were enrolled in this cross-sectional study, including 100 cases with DPN and 88 cases without. Clinical data was collected and serum levels of 25(OH)D3, glycosylated hemoglobin (HbA1c), blood lipids and hepatic and renal functions were determined in all patients. Spearman correlation analysis was used to evaluate relationship between each index and 25(OH)D3, and Logistic regression was used for statistical analysis of risk factors for DPN. Results · The median of serum 25(OH)D3 level was significantly lower in the patients with DPN (34.95 nmol/ L) than that of the patients without DPN (52.6 nmol/L)(P<0.01). Spearman correlation analysis showed that there existed a negative correlation between the level of 25(OH)D3 and age, HbA1c and fasting blood glucose (r=-0.37, -0.53, and -0.29, respectively, P<0.01). The multiple Logistical regression analysis revealed that the 25(OH)D3 level was an independent risk factor for DPN. The odds ratio of serum 25(OH)D3 was 15.5 (OR=15.50, 95% CI= 2.00～119.62) for the lowest quartile versus the highest quartile. Conclusion · The decreased level of 25(OH)D3 may increase risks for the occurrence of DPN in the elders with type 2 diabetes mellitus and monitoring the level of 25(OH)D3 contributes to early detection of DPN.