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Objectives: Our goal was to explore the outcome of the study and the feasibility for patients of the technique of anatomical pulmonary segmentectomy by uniportal video-assisted thoracic surgery (VATS)., Methods: A total of 156 consecutive patients with lung lesions who received anatomical pulmonary segmentectomy by VATS between 2015 and 2016 in our hospital were enrolled. All the subjects had high-resolution, thin-section chest computed tomography (CT) examinations with 3-dimensional reconstruction, a pulmonary function test, abdominal ultrasonography, electrocardiogram and cardiac ultrasonography. The lung lesion was localized before the operation using CT reconstruction or a hookwire to plan the operative method., Results: Uniportal VATS pulmonary segmentectomy was successfully completed in 151 (96.8%) patients. Most cases involved the right apical and apical posterior segments and the left trisegment. Only 1 patient had a right middle interior segmentectomy, left upper apical anterior segmentectomy or a right lower posteriolateral segmentectomy, respectively. There were 26 cases of benign lesions (including 17 cases of atypical hyperplasia) and 130 cases of non-small-cell lung cancer. Operation time (146 ± 56 vs 113 ± 32 min), blood loss (63 ± 17 vs 54 ± 13 ml) and complication rates (13.5% vs 5.8%) were obviously lower in 2016 compared with 2015 (P < 0.01). In contrast, the preoperative hookwire localization rate was markedly higher in 2016 compared with 2015., Conclusions: Uniportal VATS anatomical pulmonary segmentectomy is safe and feasible in clinical applications. Compared with the 2- or 3-port method, there were some technical difficulties in the early phase of the learning curve for uniportal VATS that were overcome through a period of practice.