〔Abstract〕 Objective To explore the clinical efficacy of Yiqi Tongyang Huoxue prescription combined with urokinase in the treatment of acute myocardial infarction (AMI) and the effect of serum levels of soluble myeloid trigger receptor-like transcription factor-1 (sTLT-1), high mobility group box protein B1 (HMGB1). Methods A total of 92 patients with AMI patients admitted to the 7th People's Hospital of Zhengzhou from May 2020 to April 2021 were selected and divided into a control group and an observation group by random number table method, with 46 cases in each group. Patients in the control group were treated with urokinase, while patients in the observation group were treated with Yiqi Tongyang Huoxue prescription combined with urokinase. The total effective rate, cardiac function level, serum sTLT-1 and HMGB1 levels, and incidence of adverse reactions were compared between the two groups. Results The total effective rate of the observation group was 93.48%, higher than 78.26% of the control group, and the difference was statistically significant (P < 0.05). After treatment, the TCM (traditional Chinese medicine) syndrome scores of the two groups decreased to different degrees, and the TCM syndrome score of the observation group was lower than that of the control group, and the difference was statistically significant (P < 0.05). After treatment, the left ventricular ejection fraction (LVEF) of the two groups were higher than those before treatment, and the left ventricular end-systolic diameter (LVESD) and left ventricular end-diastolic diameter (LVEDD) of the two groups were lower than those before treatment. After treatment, the LVEF of the observation group was higher than that of the control group, and the LVESD and LVEDD were lower than those of the control group, the differences were statistically significant (P < 0.05). After treatment, the levels of the serum sTLT-1 and HMGB1 of the two groups were lower than those before treatment. After treatment, the levels of the serum sTLT-1 and HMGB1 of the observation group were lower than those of the control group, and the differences were statistically significant (P < 0.05). There was no significant difference in the incidence of adverse reactions between the two groups (P > 0.05). Conclusion Yiqi Tongyang Huoxue prescription combined with urokinase in the treatment of AMI patients has a significant clinical effect, can improve the cardiac function, reduce the levels of serum sTLT-1 and HMGB1, and has high safety