Abstract: To quantitatively evaluate the hydrological regime changes in the middle and lower reaches of the Yangtze River during the past 60 years, the trend and mutation of runoff were analyzed by the Mann-Kendall nonparametric test and mean difference T test at the Yichang, Hankou and Datong gauge station from 1954 to 2016. Without considering the impact of evapotranspiration, the slope change ratio of cumulative quantity (SCRCQ) was adopted to estimate the impacts of precipitation and human activities. The hydrologic regime alteration degree of 32 hydrological indicators were finally studied by the range of variability approach (RVA).The results showed that :(1) there was no significant trend change of annual runoff in the middle and lower reaches of the Yangtze River, and the abrupt change was consistent with the impoundment time of the Three Gorges Reservoir(TGR); (2) the impoundment of the Three Gorges Reservoir mainly affects the annual variation of runoff in the middle and lower reaches of the Yangtze River, especially the annual maximum of the runoff, and the impact degree decreases with the increase of distance from the reservoir; (3) the hydrological alteration degrees of Yichang, Hankou and Datong hydrologic stations were 55%, 46% and 32% respectively, and the hydrological regime of the middle and lower reaches of the Yangtze River belonged to medium-high alteration generally; (4) the precipitation of climate change is the leading factor resulting to the change of annual runoff in the middle and lower reaches of the Yangtze River, and its contribution degree accounts for about 70%, while the contribution degree of water draw, water conservancy projects and other human activities accounts for about 30%.