In order to study the denitrification and N₂O production rate of sediments in the Dongyu River in spring, the sediment and surface water samples were collected along the Dongyu River in May, 2017. The acetylene inhibition method and incubation method were applied to determine the denitrification potential, denitrification rate and N₂O production rate. The N₂O diffusion flux was estimated by using the headspace equilibrium method and two-layered diffusion model. Moreover, the physicochemical properties of sediment and surface water were also analyzed to discuss the main influencing factors by using correlation analysis. The results showed that denitrification potential of the spring sediments in Dongyu River was 7 305.8~26 947.7 μmol/(m²·h), but from upstream to downstream, it tended to increase first and then decrease. The denitrification rate of sediment was 86.6~694.2 μmol/(m²·h), which was significantly lower than the denitrification potential at the same site (P < 0.01), moreover the change of the denitrification rate along the Dongyu River was not in accordance with the change of denitrification potential. The denitrification rate was only significantly positively correlated with the NH₄⁺ concentration in the sediments (P < 0.05), which indicated that the denitrification rate of sediment might be synthetically affected by the physicochemical properties of the sediments and surface water, and also there might be coupling effect of nitrification and denitrification in the water body. The N₂O production rate was 19.8~144.3 μmol/(m²·h), and the diffusion flux of N₂O varied from 170.9~667.8 μmol/(m²·h), which indicated that the water body of the Dongyu River was the source of N₂O emission to atmosphere. The diffusion flux of N₂O was significantly higher than the N₂O production rate of the corresponding point (P < 0.01), and it was significantly positively correlated with the concentrations of DO and NO₂^- in the overlying water (P < 0.05), which suggested that besides the N₂O produced from the sediment, the transformation of nitrogen in the overlying water was also an important pathway to discharge N₂O to atmosphere from the Dongyu River in spring.