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Abstract: Dysregulated microRNAs (miRNAs) in the plasma of patients with lung squamous cell carcinoma (LSCC) might serve as biomarkers for LSCC diagnosis. The expression of miRNAs was performed using quantitative reverse transcription–polymerase chain reaction (qRT‐PCR) on the basis of Exiqon panels in the initial screening phase including three male LSCC pool samples and one normal control (NC) pool sample (per 10 samples were pooled as one pool sample). After the training (32 LSCC vs. 31 NCs), the testing (55 LSCC vs. 55 NCs), and the external validation (15 LSCC vs. 15 NCs) stages via qRT‐PCR, a four‐miRNA signature (miR‐181a‐5p, miR‐21‐5p, miR‐106a‐5p, and miR‐93‐5p) was identified for LSCC detection. Areas under the receiver operating characteristic (ROC) curve (AUC) of the four‐miRNA panel for the training, the testing, and the external validation phases were 0.795, 0.827, and 0.914, respectively. Then, the four miRNAs were explored in LSCC tissue samples (23 LSCC vs. 23 NCs), and their expression was significantly up‐regulated. However, none of the four miRNAs found significantly up‐regulated in plasma exosomes expect miR‐93‐5p with borderline significance (16 LSCC vs. 16 NCs). In summary, our study established a four‐miRNA peripheral plasma signature, which contributed to diagnosing male LSCC patients in China to a certain degree. [ABSTRACT FROM AUTHOR]