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Aim To investigate the levels of krüppel-like factor 2 (KLF2) and endothelial nitric oxide synthase 3 (NOS3) in the serum of patients with acute cerebral infarction (ACI) of large-artery atherosclerosis (LAA) type, and to analyze their value in the diagnosis and disease assessment of LAA type ACI. Methods A total of 150 patients with LAA type ACI were divided into mild group (n = 36), moderate group (n = 48), and severe group (n = 66) based on their condition. Additionally, a control group ( n = 150) was selected from health exminers during the same period. The levels of serum KLF2 and NOS3 in each group were compared; receiver operator characteristic(ROC) curve was applied to analyze the diagnostic value of serum KLF2 and NOS3 levels for LAA type ACI and the predictive value for the occurrence of severe LAA type ACI, respectively. Results The serum KLF2 and NOS3 levels were significantly lower in LAA type ACI group than those in control group (P<0. 05). The serum KLF2 and NOS3 levels in the mild, moderate and severe groups were significantly decreased in turn(P<0. 05). The area under the curve (AUC) of the combined diagnosis of serum KLF2 and NOS3 for LAA type ACI was 0. 858, with a sensitivity of 73. 33% and a specificity of 86. 00%, which was superior to the individual diagnosis of KLF2 and NOS3 (Zcombined detection-KLF2 = 3. 796, Zcombined detection-NOS3 = 4. 689, all P< 0. 001). The AUC of combined prediction of serum KLF2 and NOS3 for the occurrence of severe LAA type ACI was 0. 878, with a sensitivity of 77. 27% and a specificity of 90. 48%, which was superior to the independent prediction of KLF2 and NOS3 (Zcombined detection-KLF2 = 2. 401, P= 0. 016; Zcombined detection-NOS3 = 3. 070, P = 0. 002). Conclusions The serum levels of KLF2 and NOS3 in patients with LAA type ACI were significantly reduced and negatively correlated with the severity of the disease. The combination of the two has high evaluation efficacy in the diagnosis and disease prediction of LAA type ACI. [ABSTRACT FROM AUTHOR]