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Objective To analyze the expression levels of serum transcription factor PU.1 and interleukin-4 (IL-4) in children with acute upper respiratory tract infectious diseases and explore their clinical diagnostic values. Methods A total of 78 children with acute upper respiratory tract infection treated in our hospital were selected as the disease group, and 78 healthy children with physical examination during the same period were selected as the normal control group. Fluorescence quantitative method and enzyme linked immunosorbent assay (ELISA) were used to detect the expression levels of serum transcription factors PU.1 and IL-4 in children respectively. The receiver operating characteristic (ROC) curve was used to analyze its clinical diagnostic value. Multivariate Logistic regression was used to analyze the independent risk factors affecting the occurrence of acute upper respiratory tract infectious diseases. Results The serum levels of interleukin-6 (IL6), C reactive protein (CRP), procalcitonin (PCT), tumor necrosis factor alpha (TNF-α), IL-4 and the relative expression of PU.1 were significantly higher in the disease group than those in the normal control group (P＜0.05). The areas under the ROC curve of serum IL-4 level and the relative expression of PU.1 and their combined detection for diagnosing the upper respiratory tract infectious diseases in children were 0.752, 0.855 and 0.984, respectively, the corresponding sensitivities were 58.97%, 78.21%, 94.87%, respectively, and the specificities were 83.33%, 78.21% and 97.44%, respectively. A multivariate Logistic regression analysis was performed, and the results showed that higher serum IL-4, PU.1, IL-6, CRP and TNF-α expression levels were independent risk factors affecting the occurrence of upper respiratory tract infectious diseases (P＜0.05). Conclusion The serum levels of PU. 1 and IL-4 in children with acute upper respiratory tract infectious diseases are relatively high, which have certain diagnostic value. [ABSTRACT FROM AUTHOR]