Objective It has been an established practice abroad to assist intravenous thrombolytic therapy for cerebral infarction (CI) with the telestroke program in primary hospitals, but there are rare relevant reports in China. This study is based on a 7×24 telestroke program jointly established by the Stroke Center of Shanghai Ninth People's Hospital (Shanghai Center) and the Stroke Center of the People's Hospital of Xiangyun in Yunnan Province, China (Xiangyun Center). Xiangyun Center is supported by Shanghai Center in intravenous thrombolytic therapy for CI.Methods The cases of acute CI within 1 year were prospectively collected from the telestroke program of Shanghai Center and Xiangyun Center, and were summarized for the quality control and follow-up of the cases undergoing an intravenous thrombolysis; the two centers were compared in terms of intravenous thrombolysis rate, intravenous thrombolysis rate within 3 hours, door-to-needle (DTN) time, onset-to-treatment (OTT) time, and 3-month follow-up results of the following:modified Rankin Scale (mRS) score, bleeding rate, and mortality.Results Within the 1-year telestroke observation period, there was a significant difference in intravenous thrombolysis rate between the two centers (27.04% for Shanghai Center vs 7.73% for Xiangyun Center, P=0.000), but there were no significant differences observed between the two centers in DTN time (62.76±26.41 min for Shanghai Center vs 70.55±28.51 min for Xiangyun Center, P>0.05) or 3-month follow-up results of mRS score, bleeding rate, and mortality (P>0.05).Conclusions The prognosis and safety of intravenous thrombolytic therapy under telestroke assistance in Xiangyun Center is not inferior to those of Shanghai Center. Telestroke is an effective method to improve the intravenous thrombolysis rate in primary hospitals in rural areas.