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High-resolution ultrasonography was used to analyze the nerve cross-sectional area (CSA) of the median nerve at 7 sites: the wrist crease, pisiform bone, hamate bone, 6 cm proximal to the tip of the wrist crease, proximal forearm (where the nerve enters the pronator teres muscle), 4 cm proximal to the tip of the medial epicondyle, and mid-humerus (mid-point between elbow crease and axilla) in 200 healthy volunteers from Guiyang, China. Results showed similar CSA values between the left and right sides, but the CSA 6 cm proximal to the tip of the wrist crease, proximal forearm, 4 cm proximal to tip of the medial epicondyle, and mid-humerus in males was greater than that of females. Moreover, CSA values at the wrist crease, pisiform bone, and hamate bone were greater in the middle-aged and old groups when compared to the young group, and correlated with body mass and height. Thus, reference values of median nerve CSA of the upper limbs can facilitate the analysis of abnormal nerve conditions.