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Objective: To investigate the clinical value and application of ATP based bioluminescence tumor chemosensitivity assay(ATP-TCA) in the chemotherapy for hydrothorax caused by non-small cell lung cancer (NSCLC).Methods: Hydrothorax specimens from 120 NSCLC patients were analyzed by ATP-TCA and the most sensitive chemotherapeutic drugs were used in NSCLC patients (treatment group).At the same time,56 NSCLC patients with hydrothorax were admitted in our department and were given chemotherapy without guidance of the ATP-TCA(control group).Before the third chemotherapeutic cycle, clinical outcomes were analyzed in the two groups.Results: Effective rate of hydrothorax in treatment group was 67%, while 46% in control group (P<0.05).In refractory hydrothorax patients, it was 69% and 40% (P<0.05), respectively.In vitro results correlated well with clinical outcomes (P<0.01).Conclusion: Effective rate of chemotherapy for hydrothorax in NSCLC is higher in treatment group than that in control group.ATP-TCA is especially helpful for refractory hydrothorax.