Objective To investigate the value of EBUS-TBNA in the diagnosis of chest diseases. Methods Retrospective analysis of 343 patients examined by EBUS-TBNA from September 2009 to August 2011, 219 males and 124 females; age (59.4±13.6) years. Among them, 208 patients had suspicious or definite lung cancer with mediastinal or hilar lymph node enlargement on imaging; 94 patients had mediastinal and/or hilar lymph node enlargement or mediastinal occupancy without intrapulmonary occupancy; and 41 patients had intrapulmonary parenchymal occupancy adjacent to the airway. Results Patients with short chest lesion diameter (1.94±1.01) cm, punctured chest lesions (1.77±0.86) per case, and punctured (4.71±2.69) times, with an average of 2.66 punctures per lesion. The overall sensitivity of EBUS-TBNA for the diagnosis of benign and malignant lesions in the mediastinum and/or hilum was 95.6% (238/249 cases), specificity was 100% ( 94/94 cases), positive predictive value was 100% (238/238 cases), negative predictive value was 89.5% (94/105 cases), and accuracy was 96.8% (332/343 cases).208 patients with definite or Among the 208 patients with definite or suspected lung cancer, 151 were diagnosed with mediastinal lymph node metastasis by EBUS-TBNA, 4 with tuberculosis, and 2 with stage II nodal disease; 37 of the 51 patients with negative EBUS-TBNA underwent further surgery, and 32 were confirmed to be true negative. The sensitivity of EBUS-TBNA was 96.8% (151/156 cases), specificity 100.0% (32/32 cases), accuracy 97.3% (183/188 cases), positive predictive value 100.0% (151/151 cases), negative predictive value 86.5% (32/37 cases). 22 of 94 patients with mediastinal and/or hilar lymph node enlargement or mediastinal occupancy were malignant lesions, and 23 of 73 benign lesions were diagnosed as lymph node The sensitivity of EBUS-TBNA in diagnosing benign and malignant mediastinal lesions in this group was 88.0% (22/25 cases), specificity 100% (73/73 cases), and negative predictive value 95.9% (70/73 cases) accuracy 97.9% (92/94 cases). The accuracy of EBUSTBNA for the diagnosis of malignant and benign lesions in this group was 88.0% (22/25 cases) and 95.9% (70/73 cases), respectively. 33 of the 41 cases of intrapulmonary parenchymal occupations adjacent to the large airways were malignant; 8 cases were negative for EBUS-TBNA, 4 of which were confirmed as false negatives by further surgery. The sensitivity of EBUS-TBNA was 89.2% (33/37 cases) and the accuracy was 90.2% (37/41 cases). No puncture-related complications occurred in all cases. Conclusion EBUS-TBNA is reliable for the pathological staging of lung cancer and safe and effective for the diagnosis of thoracic diseases such as hilar or mediastinal occupations adjacent to the large airways.