To evaluate the safety and efficacy in terms of infarct-related artery (IRA) patency of prourokinase (proUK) injection within 6 h after symptom onset in patients with ST-elevation myocardial infarction (STEMI). A total of 1851 patients with acute myocardial infarction were enrolled to receive thrombolytic therapy with proUK. Qualifying patients were injected with 20 mg proUK intravenously followed by intravenous infusion of 30 mg proUK within 30 min. IRA was evaluated by coronary angiography at 120 min after thrombolysis. The efficacy and safety of proUK thrombolysis were analyzed in all the enrolled patients. Successful clinical reperfusion was observed in 1580 patients (85.4%). The number of leads with ST segment elevation and the amplitude of ST segment elevation were significantly reduced within 24 h after thrombolytic therapy with proUK. Eighty-three patients (4.48%) had died by the end of follow-up (7 days after thrombolysis), and the incidences of post-infarction angina, re-infarction or reocclusion, and emergency revascularization were 15.2, 4.21, and 8.27%, respectively. The most common bleeding complication was subcutaneous or mucosal bleeding and the incidence of intracranial hemorrhage was as lower as 0.32%. The fibrinolysis therapy with proUK is efficient and safe with very low rate of bleeding complications. It provides an alternative treatment options for STEMI patients especially in settings where primary PCI cannot be offered in a timely manner.