Objective To investigation the distribution and antimicrobial resistance of clinical bacterial strains from Xiangya Hospital of Central South University in 2013-2017, and provide basis for clinical rational use of antimicrobial agents. Methods Strains isolated from cultured specimens of outpatients and inpatients were collected, antimicrobial susceptibility testing was performed by Kirby-Bauer method or automated system, bacterial resistance monitoring data were analyzed statistically. Results A total of 38 075 bacterial strains were isolated, including 13 184 (34.6%) strains of gram-positive bacteria and 24 891 (65.4%) strains of gram-negative bacteria. The top five isolated bacteria were Escherichia coli(n=5 158,13.5%),Acinetobacter spp.(n=4 740,12.4%), Klebsiella spp. (n=4 470,11.7%), coagulase-negative staphylococcus (n=4 008,10.5%), and Pseudomonas aeruginosa(n=3 577, 9.4%). Isolation rates of methicillin-resistant Staphylococcus aureus(MRSA) and methicillin-resistant coagu- lase-negative staphylococcus (MRCNS) were 31.7% (980/3 096) and 77.7% (3 113/4 008) respectively, isolation rate of MRSA showed a decreasing tendency(P<0.01); resistance rate of methicillin-resistant strains of Staphylococcus was higher than methicillin-sensitive strains; vancomycin-resistant Staphylococcus and linezolid-resistant Staphylococcus aureus were not found. Resistance rates of Enterococcus faecium to penicillin, ampicillin, high concentration gentamycin, high concentration streptomycin, erythromycin, nitrofurantoin and fluoroquinolones were all much higher than those of Enterococcus faecalis (all P<0.01), resistance rate of Enterococcus faecalis and Enterococcus faecium to vancomycin and linezolid was<3.5%. Linezolid- or vancomycin-resistant Streptococcus spp. was not found. Resistance rates of Escherichia coli and Klebsiella pneumoniae to carbapenems were 0.6%-3.9% and 6.3%-24.9% respectively. Resistance rates of Pseudomonas aeruginosa and Acinetobacter baumannii to imipenem were 29.5%-34.7% and 69.9%-85.7% respectively. Resistance rates of Escherichia coli, Klebsiella pneumoniae and Acinetobacter baumannii to carbapenems increased year by year (all P<0.01). Conclusion Resistance rate of Acinetobacter baumannii to carbapenems is high, resistance rate of Klebsiella pneumoniae to carbapenems is rising rapidly. It is necessary to strengthen the rational use of antimicrobial agents, take effective measures to control healthcare-associated infection, and reduce the emergence and spread of antimicrobial-resistant bacteria.