The release pathway of dissolved organic matter (DOM) in sediments of the Xinkaihe River in Shenyang, as well as the effect of time, temperature, pH, sediment particle size, salinity, and dissolved organic carbon (DOC) concentration in the overlying water on the release of DOM in sediments was evaluated. In addition, the effect of release of DOM in sediments on the spectroscopy characteristics of DOM in the overlying water was also examined. The results showed that the mixing action of interstitial water had the greatest influence on the release of DOM in sediments, which was followed by the static release of surface sediment, while the release of suspended particles made slight influence on the release of DOM in sediments. The release amount of DOM in sediments increased with the increase of temperature, pH, sediment particle size, and the salinity. When the DOC concentration in the overlying water was relatively high, the DOM moved from the overlying water to the sediment. The disturbance could affect the migration of DOM in sediments. The fulvic acid- and aromatic protein-like fluorophores were dominant in fluorescent materials in DOM in sediments. The fluorescent materials with excitation wavelength of 280~300nm and 320~380nm were the main fluorescent materials which were released from the sediment to the overlying water. Moreover, the disturbance promoted the release of fluorescent materials from the sediment to the overlying water.