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OBJECTIVE:To investigate the effects on the brain using three needling manipulations (twirling,lifting-thrusting,and twirling plus lifting-thrusting)when the right-side Zusanli (ST 36) acupoint was stimulated with needles.METHODS:Seventeen healthy subjects accepted three needling manipulations stimulating the right Zusanli (ST 36) over separate days.Functional magnetic resonance imaging was used to detect changes in the brain during the manipulations,and then the needling sensations were recorded using the MGH acupuncture sensation scale (MASS) after each scan.fMRI data were processed using Statistical Parametric Mapping 8 to analyze the positive and negative activation in the brain induced by different acupuncture manipulations.RESULTS:The individual needling sensations showed no statistically significant differences among the three manipulations.However,the MASS index showed that lifting-thrusting ＞ twirling plus lifting-thrusting ＞ twirling.Lifting-thrusting activated left premotor cortex,left postcentral,right middle frontal gyrus,left inferior frontal gyrus,right lingual gyrus,left insula,right putamen,bilateral cingulate gyrus and right cerebellum;and deactivated bilateral hippocampus and left caudate.Twirling activated bilateral orbital middle frontal gyrus,left opercular and triangular inferior frontal gyrus,and right middle occipital gyrus;and deactivated bilateral precuneus,right amygdala,left anterior cingulate gyrus,right inferior temporal gyrus,right middle frontal gyrus,right supplementary motor gyrus,and left postcentral.Twirling plus lifting-thrusting activated bilateral postcentral,left inferior occipital gyrus,left insula,left thalamus,left cingulate gyrus,and right putamen;and deactivated right superior frontal gyrus,right superior parietal gyrus,right temporal gyrus,right middle occipital gyrus,right insula,and left lingual gyrus.Pairwise comparisons of the three manipulations showed that signals induced by lifting-thrusting were the strongest,especially in the limbic system,followed by twirling plus lifting-thrusting;twirling alone was the weakest.CONCLUSION:Three methods of needling manipulations similarly activated areas associated with the somatosensory system,vision,cognition,and emotional regulation.This may have significant implications for acupuncture in clinical practice.