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Background: The association between loop diuretics and acute kidney injury (AKI) remains unclear. Methods: The population studied was selected from the Epidemiology of AKI in Chinese Hospitalized patients (EACH) study. Exposure to loop diuretics was defined as any filled prescription prior to the date when AKI was detected in patients with HA-AKI, and prior to the last date of SCr testing in those without AKI. The outcome was AKI, defined by the Kidney Disease Improving Global Outcomes criteria. Associations between loop diuretics and HA-AKI were examined by Cox proportional hazards models adjusted for baseline and time-dependent covariates. Results: Of the 150,020 patients, 16,437 (11.0%) were prescribed loop diuretics, and 5717 (3.8%) experienced HA-AKI events. The crude rates of HA-AKI in patients who were and were not prescribed loop diuretics were 1632 (9.9%) and 3262 (2.8%), respectively. A multivariate cox proportional hazards analysis showed that exposure to loop diuretics was associated with significantly increased risks of HA-AKI compared with non-users (hazard ratio (HR), 1.61; 95% CI (confidence interval), 1.55–1.67), other diuretics (HR, 1.09; 95% CI, 1.03–1.15), and osmotic diuretics (HR, 1.30; 95% CI, 1.20–1.42). Compared with other diuretics, the use of loop diuretics was associated with higher risks of HA-AKI in women, in patients without hypertension, in patients without heart failure, in patients without liver cirrhosis, and in patients not requiring surgery. Conclusions: Loop diuretics are widely used and are associated with increased risks of HA-AKI in hospitalized adults. Renal function should be more closely monitored during the use of loop diuretics.