Objective To investigate the effect of Clostridium Butyricum Powder combined with interferon in the treatment of children with Rotavirus (RV) infectious diarrhea and its influence on the levels of lactic acid and inflammatory factors. Methods A total of 86 children with RV infectious diarrhea who were admitted in Heze Maternal and Child Health Hospital from December 2020 to November 2021 were selected as the research objects. They were divided into the observation group (43 cases) and the control group (43 cases) according to random number table method.The control group was treated with interferon, and the observation group was treated with Clostridium Butyricum Powder combined interferon. The clinical efficacy, the levels of lactic acid and inflammatory factors were compared between the two groups. Results The total effective rate of treatment in the observation group was higher than that in the control group, and the difference was statistically significant (P＜0.05). After treatment, the levels of D-lactic acid (D-LAC) in the two groups were lower than those before treatment, and the levels of D-LAC after treatment in the observation group were lower than those in the control group, and the differences were statistically significant (P＜0.05). After treatment, the levels of interleukin-6 (IL-6), interleukin-10 (IL-10) and tumor necrosis factor-α (TNF-α) in the two groups were lower than those before treatment, and the levels of IL-6, IL-10, TNF-α after treatment in the observation group were lower than those in the control group, and the differences were statistically significant (P＜0.05). Conclusion The treatment of children with RV infectious diarrhea by Clostridium Butyricum Powder combined with interferon can improve the clinical efficacy and improve the levels of lactic acid and inflammatory factors.