Abstract: Climate warming, population growth and excessive economic development have exacerbated water scarcity in China. The spatio-temporal evolution of China’s human water use (341 prefectures) is explored, based on the methods of Breaks for Additive Seasonal and Trend and Spatial Autocorrelation Analysis. Results show that China’s human water usage was continuously increasing from 1965 to 2013, although a breakpoint was detected mainly after 1990. Increasing and decreasing of water use coexisted after the breakpoint year, thus resulting in a deceleration of China’s human water use. The distribution of human water use had spatial agglomeration characteristics, and the agglomeration of water use was high in the east and low in the west (except Xinjiang) basically in line with Hu Huanyong boundary. The agglomeration regions of human water use and irrigation water use were relatively consistent before 1995, and regions with high water use concentrations were in Xinjiang and Yangtze River Delta. But the agglomeration regions of human water use were more similar to the agglomeration regions of industrial water use after 1995. Regions with high concentrations of industrial and urban and rural water uses were distributed in northeastern China, northern China and southern China, and regions with low concentrations were in western China (largest agglomeration effect existed in rural water use). Exploring the spatio-temporal characteristics of water use in China could provide a scientific basis for water resources management and the improvement of water use efficiency.