Abstract: OBJECTIVE To explore the evolution of the resistance of common gram-negative bacteria in hospitals to quinolones from 2009 to 2014, so as to guide the clinical use of quinolone antibacterial agents. METHODS The evolution of the resistance of common gram-negative bacteria isolated from clinical specimens of a large top three hospital a in Guangxi province from 2009 to 2014 was discussed by statistics and analysis. The data were analyzed by SPSS 18.0. RESULTS Among seven common gram-negative bacteria in hospital, Escherichia coli and Acinetobacter showed the highest drug resistance rates: for six consecutive years, the drug resistance rates of E. coli to ciprofloxacin and levofloxacin were above 60%, while those of Acinetobacter to ciprofloxacin and levofloxacin were above 58.9%. CONCLUSION The situation of the resistance of gram-negative bacteria in hospital to ciprofloxacin and levofloxacin is grim, and E. coli and Acinetobacter face the most formidable challenge. Experience-directed medication of clinicians should refer to the bacterial resistance surveillance data of this hospital and maximize the effects of quinolone drugs.