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Hydrologic regimes are increasingly altered under the impacts of climate change and human activities. Streamflow data from 1960 to 2014 were analysed to investigate changes in the flow regimes in the Yangtze River using multiple hydrologic metrics and the Budyko framework. The long-term data were separated into two periods: the preimpact period (1960-2002) and the postimpact period (2003-2014), according to the year the Three Gorges Dam began operation. The results showed that both indicators of hydrologic alteration and ecoflow metrics were clearly altered. The highly changed indicators included flow in February, annual minimum 1-, 3-, 7-, 30-, and 90-day flows, base flow index, date of annual minimum flow, and low pulse duration. The integrated degree of hydrologic alteration ranged from 41% to 61%, indicating a moderate alteration of the flow regimes in the Yangtze River. The regulation of the Three Gorges Dam increased low flow and weakened peak flow, which resulted in autumn ecodeficit and winter ecosurplus increasing dramatically since the 2000s. The ecoflow metrics were more sensitive to precipitation than to potential evapotranspiration. The joint effects of human activities and climate change varied among the river reaches in the different decades. The streamflow was mainly affected by human activities in the upper reach during the 1970s-1990s, with a contribution ratio ranging from 63% to 77%. Climate change shifted to a major contributor in the middle and lower reaches since the 1980s as well as in the upper reach in 2000-2014, accounting for 50-82% of the streamflow changes. These different responses were primarily caused by the variations of precipitation and intensive human activities, particularly the rapid growth of reservoirs and other large projects since the 1970s in the upper Yangtze River. These results provide interesting insights into the spatio-temporal hydrologic alteration across the Yangtze River.