Objective To analyze the distribution characteristics and clinical significance of peripheral blood lymphocyte subsets in patients with middle-advanced non-small cell lung cancer(NSCLC).Methods Ninety-three middle-advanced NSCLC patients admitted to our hospital from January 2022 to April 2023 were studied.The proportions and absolute values of peripheral total T cells,CD4+T cells,CD8+T cells,total B cells and natural killer(NK)cells were measured by flow cytometry,and their distribution characteristics were analyzed.Results Blood routine examination showed that 58.06%of total patients had a lower percentage of peripheral blood lymphocytes,and 35.48%had a lower absolute value of peripheral blood lymphocytes.Flow cytometry analysis showed that the distribution of T(84.95%),B(76.34%),and NK(92.47%)cells in peripheral blood was mostly within the normal range.The abnormal CD4+/CD8+ratio was found in 60.22%of total patients.The absolute values of peripheral blood T(60.22%),CD4+T cells(58.06%),and CD8+T cells(64.52%)were mainly reduced.44.09%of patients showed a decrease in absolute values of B cells and 32.26%had decreased absolute values of NK cells.The proportion of total T cells in peripheral blood of adenocarcinoma patients was significantly lower than that of squamous cell carcinoma patients(P=0.047).The proportion and absolute value of NK cells in peripheral blood of adenocarcinoma patients were significantly lower than those of squamous cell carcinoma patients(P=0.0025 and 0.02,respectively).There was no significant difference in the distribution of other factors among different pathological types(all P>0.05).Conclusion Patients with middle-advanced NSCLC have decreased peripheral blood immune cells and imbalanced CD4+/CD8+ratio.NK cell number is correlated with the pathological type of advanced NSCLC patients.