The aim of this study was to evaluate the application of phase-contrast cine magnetic resonance imaging (MRI) in endoscopic aqueductoplasty (EA) for patients with obstructive hydrocephalus. The clinical diagnosis of hydrocephalus caused by aqueduct obstruction in 23 patients was confirmed by phase-contrast cine MRI examination. The patients were treated with EA and MRI was repeated during the follow-up. The cerebrospinal fluid (CSF) flow velocity in the aqueduct was measured to determine whether the aqueduct was obstructed. The results of phase-contrast cine MRI examinations indicated that there was no CSF flow in the aqueduct for all patients prior to surgery. Aqueductoplasty was successfully performed in all patients. The results of phase-contrast cine MRI examinations performed a week after surgery demonstrated an average CSF flow velocity of 4.74±1.77 cm/sec. During the follow-up, intracranial hypertension recurred in two patients in whom CSF flow was not observed in the aqueduct by the phase-contrast cine MRI scan. Aqueduct re-occlusion was revealed by an endoscopic exploration. By measuring the CSF flow velocity, phase-contrast cine MRI accurately identifies aqueduct obstruction. Cine MRI is a nontraumatic, simple and reliable method for determining whether the aqueduct is successfully opened following aqueductoplasty.