Objective Increasing neuroimaging studies have focused on the sustained after effects of acupuncture, especially for the changes of brain activities in rest.However, short-period stimuli have mostly been chosen in these works.The present study attempted to investigate how resting state brain activities in healthy subjects were modulated by relatively long-period (30 min) acupuncture, a widely used modality for clinical practice.Methods Transcutaneous electric acupoint stimulation (TEAS) or intermittent minimal TEAS (MTEAS) were given for 30 min to 10 subjects with at least 3 months apart.Functional MRI(fMRI) data were collected including the pre-stimulation resting state and post-stimulation resting state, using arterial spin labeling (ASL) techniques, representing both cerebral blood flow (CBF) signals and blood oxygen-dependent level (BOLD) signals simultaneously.Results (1) While mean global CBF was not altered by TEAS or MTEAS, pain-processing, decision making and impulsive related brain regions (primary and secondary somatosensory cortex, posterior insula and lateral prefrontal cortex) showed more deactivation after 30 min TEAS as compared to MTEAS.(2) Analysis of BOLD signals revealed that TEAS, but not MTEAS, produced an increased functional connection between posterior cingulate cortex and anterior cingulate cortex, a brain region involved in anti-nociceptive, attention and reward.Conclusion Our results implied that modulation of these regional brain activities and brain network may relate to acupuncture analgesia as well as other acupuncture-associated therapeutic effects.Furthermore, the resting state regional CBF quantified by ASL perfusion fMRI may serve as a potential biomarker in future acupuncture studies.