Objective: To study the application value and application path of pathological examination in gynecological physical examination. Methods: A total of 1200 patients undergoing gynecological physical examination in X Hospital from January 2024 to December 2024 were selected. All patients received cervical Pap smear examination, and patients with abnormal examination results underwent colposcopic biopsy and HPV infection test for cervical cancer screening. Results: The results of cervical Pap smear showed that a total of 780 patients among 1200 patients showed cervical abnormalities. The Pap smear combined with colposcopy was used for pathological examination to detect cervical cancer lesions in time, and biopsy and HPV infection examination were arranged for high-risk patients to provide data reference for clinical treatment. Conclusions: Pathological examination has a good screening effect in gynecological physical examination. It can detect lesions early and take timely intervention measures, which is helpful to reduce the incidence and mortality of the disease.