Objective:To observe and investigate the response of different twist manipulation on whole-brain functional connectivity based on functional integration perspective.Methods:The 40 healthy subjects were randomly divided into the control group and the treatment group.The treatment group was further divided into the 120 r/min group,the 180 r/min group and the 240 r/min group according to different acupuncture manipulation,10 cases in each group.Each group was manipulated according to different stimulus parameters,but the control group only retained the needle without any manipulation.VAS was utilized to evaluate acupuncture sensation,and the difference of whole-brain functional connectivity was compared based on fMRI and ICA.Results:The VAS results showed that other sensations of acid,swelling,numbness and pain were significantly different in each group(P<0.01)except heaviness feeling.The ICA results revealed that after manipulation,the strength of whole-brain functional connectivity in the superior orbitofrontal gyrus was decreased in the 120 r/min group,the strength of whole-brain functional connectivity in the medial frontal gyrus was decreased in the 180 r/min group,and the strength of whole-brain functional connectivity in the middle temporal gyrus was increased in the 240 r/min group.There was no significantly change in the control group.Compared with the control group,the 120 r/min group showed that the strength of whole-brain functional connectivity in the calcarine was decreased,the 180 r/min group showed that the strength of whole-brain functional connectivity in the paracentral lobule was increased,and the 240 r/min group showed that the strength of whole-brain functional connectivity in the superior temporal gyrus,precuneus and superior frontal gyrus were decreased.Conclusion:Various twist manipulation have different responses on acupuncture sensation and whole-brain functional connectivity in subjects,which preliminarily predicts that different twist manipulation might produce different effects by mediating neural functional activity in various brain regions.