Objective A high-performance liquid chromatography(HPLC)method was modified and used to determine the inhibitory effects of GM6001 and inhibitor A on tumor necrosis factor-α converting enzyme(TACE).Methods TACE and polypeptides substrate were incubated for 15 min at 37°C.Ala-Dpa was added as internal standard of quantitative analysis.Then the solution was analyzed by HPLC.The 55% methanol aqueous solution was used as the mobile phase.The wavelength of detector was 353 nm.The ratio of the peak area of remaining substrate to that of internal standard was determined.And the amounts of inverted substrate could be obtained from calibration curve.The TACE activity could be calculated.Results The relative peak areas of substrate were linearly increased depending on the growth of substrate concentration.The correlation coefficient was 0.996 8 and linear range was from 10 to 400 μmol/L.Precision experiments indicated that the precision was improved obviously by using internal standard method in the determination of TACE activity by HPLC.The values of 50% inhibitory concentration IC50 of GM6001 and inhibitor A determined by the newly proposed method were 317.5 and 175.8 nmol/L,respectively.Conclusion The HPLC method assaying TACE activity with Ala-Dpa as internal standard is more accurate,and more practical for screening of TACE inhibitors.

Background and objective China is a high-incidence area of lung cancer,and its morbidity and mortality were the highest in malignant tumors.At present,the popularization of low-dose computed tomography (CT) examination has significantly improved the detection rate of early lung cancer,anatomical segmentectomy is currently widely used early in non-small cell lung cancer (NSCLC) and those who cannot tolerate lobectomy in patients with lung cancer.However,the anatomical structure of segment and its surgical operation is relatively complex,lead to segmentectomy has a high risk and difficulty.We performed anatomical segmentectomy by use of combining three-dimensional computed tomography bronchography and angiography (3D-CTBA) and three-dimensional video-assisted thoracic surgery (3D-VATS) single-operationhole minimally invasive surgery in the treatment of NSCLC to investigate its clinical effect,and evaluate its clinical relevant feasibility and theoretical basis.Methods We carried out a retrospective review of the 57 cases by use of combining 3D-CTA in preoperative and 3D-VATS single-operation-hole anatomical segmentectomy in intraoperative in the treatment of NSCLC performed in Department of Thoracic Surgery,the First Affiliated Hospital of Soochow University from October 2015 to April 2017.Results The whole group was successfully performed by use of VATS without anyone conversion to thoracotomy.The mean operation time was (142.2±28.3) min,and the mean blood loss was (93.8±46.5) mL.The mean number of lymph node dissection was (9.1±2.2),and the mean postoperative thoracic drainage was (429.8±181.2) mL.The postoperative retention of chest tube time was (2.8±1.1) d.The mean hospitalization time was (5.2±1.3) d.The postoperative pathology showed 9 cases with benign lesions,accounting for 15.7％,48 cases with malignant lesions,accounting for 84.2％.Postoperative complications:pulmonary infection in 3 cases (5.2％),atelectasis in 1 case (1.9％),small amount of hemoptysis in 1 case (1.9％),lung leakage ＞3 d in 2 cases (3.5％).Arrhythmia in 4 cases (7.0％).The patients were followed up for 10 months.No complications occurred such as bronchial pleural fistula,chylothorax,encapsulated pleural effusion and those patients were followed up without recurrence and distant metastasis.Conclusion The use of combining 3D-CTBA and 3D-VATS single-operation-hole to anatomical segmentectomy is safe and effective in the treatment of NSCLC and is suitable for early non-small cell lung cancer,especially those can not tolerate lobectomy.