Add info. Bladder cancer (BC) is the most common urologic malignancy, facing enormous diagnostic challenges.Urinary extracellular vesicles (EVs) is a particularly promising source for developing diagnostic markers for bladder cancer because of the direct contact between urine and bladder. We investigate biomarkers for bladder cancer diagnosis based on an N-glycomics study involving the clinical cohort of 333 individuals. Finally, eight N-glycans were screened out to construct a diagnostic model, able to well differentiate bladder cancer from healthy individuals.To further understand the functions and potential roles of urinary EVs, we conducted N-glycoproteomic analysis of urinary EVs from the BC, UB, UM, and HC groups to explore the nature of glycoproteins carrying cancer-associated N-glycan signatures in EVs.