Pseudomonas aeruginosa is a major pathogen responsible for nosocomial infections. A 16-year retrospective report from 2000 to 2015 was conducted to assess the antimicrobial resistance of P. aeruginosa in Southern China. A total of 1387 P. aeruginosa were collected from inpatients and outpatients. Susceptibility testing results were interpreted according to the Clinical and Laboratory Standards Institute (CLSI, 2015). Piperacillin, piperacillin-tazobactam, ceftazidime, aminoglycosides and carbapenems remained to be active against P. aeruginosa, with resistance rates ranging from 5.6% to 29.7%. Generally, ampicillin, ampicillin-sulbactam, ceftriaxone and trimethoprim-sulfamethoxazole nearly lost the effect on P. aeruginosa, as the resistance rates increase up to 90%. Notably, sputum and blood specimen showed higher resistance rates than other sources in carbapenems, suggesting more caution should be paid on the choice of antibiotic against infections associated with respiratory tract.