Both sensitivity-based method and simulation method are used to analyze the streamflow response to climate variability and human activities in the upper catchment of the Yellow River Basin (UYRB) in this study. The separation regime of effects from climate variability and human activities is investigated. Results show that the changes of streamflow are more sensitive to precipitation than potential evapotranspiration (PET). Effect of climate variability on streamflow estimated using the sensitivity-based method is weak in the upper catchment of Jimai station, and strong in the upper catchment of Lanzhou station, where the climate effects accounted for about 50% of total streamflow changes. Effects of human activities on streamflow accounted for about 40% in the UYRB, with weaker effects in the upper catchment of Tangnaihai station than those in the upper catchment of Lanzhou station. Both climate variability and human activities are main factors to affect the changes of streamflow in the UYRB.