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Gene expression analysis was performed via qPCR using the SYBR-Green qPCR Super Mix-UDG kit on a Mastercycler ep realplex detection system (Agilent, USA)

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Neutrophils were removed from co-culture with cancer cells after 3 h of incubation at 37 degrees, washed, counted and 50,000 neutrophils re-suspended in 50 l 1X PBS, and placed in a black 96 well plate (Corning 3904), with each condition performed in triplicate

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From a clinical perspective, these results suggest that nebulized levofloxacin is a safe and effective therapeutic option for patients with CBI and chronic structural lung diseases, especially NCFB, particularly those with frequent exacerbations and prior treatment with other IAs