This study aimed to explore the feasible gender differences and similarities in cerebral activity response to the acupuncture at local acupoints around knee.Fifteen male and 15 female healthy adults were recruited and included in this study. Functional magnetic resonance imaging (fMRI) was applied to measure cerebral activity response to acupuncture at Liangqiu (ST34), Xuehai (SP-10), Neixiyan (EX-LE4), and Dubi (ST-35).Acupuncture activated the postcentral gyrus, precuneus, temporal, posterior lobe, and occipital lobe in both males and females. When compared with females, males showed brain activation in the right middle frontal gyrus, inferior frontal gyrus, right precuneus, right superior parietal lobule, left cerebellum anterior lobe; and brain deactivation in the right frontal. When compared with males, females were observed brain activation in the right frontal lobe, right parietal lobe, and right middle temporal gyrus; and brain deactivation in the left and right medial frontal gyrus.The results of this study demonstrated that the neural effects of local acupoints around knee might be different between male and female subjects. Further clinical trials should take this gender effect into account in their design of studies.