The textile printing industry discharges large volumes of effluent containing high concentrations of urea and nitrogenous compounds. Anoxic-oxic (AO) treatment is a promising method for treating printing wastewater. However, the effect of sodium sulfate (Na2SO4) salinity on the urea hydrolysis and nitrogen removal simultaneously in the AO process has received little attention. In this study, five batch reactors were used to treat synthetic printing wastewater with high urea and nitrogen concentrations. A strategy was applied to increase the Na2SO4 concentration from 0 to 19 g/L in the anoxic stage of each reactor. The effect of Na2SO4 on urea hydrolysis, total nitrogen removal and COD removal, sludge characteristics, and bacterial community structure were investigated. The findings showed that urea hydrolysis increased with increasing Na2SO4 concentration. The main mechanism of urea removal was intracellular hydrolysis, with a urea removal efficiency (URE%) of approximately 98% in all batch reactors. In addition, under the stress of Na2SO4, the total nitrogen and COD removal performances were partially inhibited. The most significant removal performances after AO treatment were observed at 0 g/L Na2SO4, with nitrogen and COD removal efficiencies of 88% and 95%, respectively. When Na2SO4 concentration reached 19 g/L, the sludge settling performance and compactness were enhanced. The extracellular polymeric substance (EPS) components in the sludge were dependent on their ability of removing organics. Bacterial community diversity analysis revealed that the enrichment of the Proteobacteria, Firmicutes, and Gemmatimonadota phyla in the anoxic stages of batch reactors was related to intracellular urea hydrolysis. Bacteriodota and Chloroflexi were responsible for total nitrogen removal in all anoxic and oxic stages. This research will develop the understanding of Na2SO4 salinity impact on simultaneous urea hydrolysis and nitrogen removal during AO treatment process.