Objective: To explore the diagnostic value of high frequency ultrasound in detecting the atrophy of extensor digitorum brevis (EDB) in patients with different course of diabetic peripheral neuropathy. Methods: The EDB of 53 DPN patients, 49 non-DPN patients and 52 healthy volunteers were scanned by high frequency ultrasound to observe the internal echo, measure the maximum thickness and cross-sectional area. The patients in group DPN were divided into three groups according to the course of disease: 5–10 years, 11–15 years, and >15 years, compared the ultrasonic images of the of patients with different courses of disease, and observed the differences between the thickness and CSA. Results: In the normal control group, the shapes of extensor digitorum brevis were plump. The thickness and cross-sectional area of control group were larger than those of DPN group and non-DPN group. The difference was statistically significant (P<0.05). In DPN patients with different courses of disease, the thickness and CSA of the EDB in the group of >15 years were significantly lower than those in the group of 11–15 years, and the thickness and CSA in the group of 11–15 years were significantly lower than those in the group of 5–10 years, differences were statistically significant (P<0.05). Conclusion: High frequency ultrasound can show the changes of the ultrasonogram of extensor digitorum brevis. It can measure the thickness and cross-sectional area of the muscle to reflect the atrophy of EDB in DPN patients with different courses of disease. It has certain diagnostic value and provides the corresponding imaging basis for the early diagnosis and early treatment of clinical early detection.