Objective To investigate the change in distribution and antimicrobial resistance of bacteria isolated from urine specimens in China. Methods According to the technical program of China Antimicrobial Resistance Surveillance System (CARSS), data of bacteria isolated from urine specimens and antimicrobial susceptibility testing results reported by all CARSS member hospitals from 2014 to 2019 were analyzed by WHONET5.6 software. Results The top 5 bacteria isolated from urine specimens of male patients were Escherichia coli (33.1%-34.6%), Enterococcus faecalis (9.2%-10.2%), Klebsiella pneumoniae (9.0%-9.4%), Enterococcus faecium (7.8%-10.2%) and Pseudomonas aeruginosa (5.6%-6.9%), the top 5 bacteria isolated from urine specimens of female patients were Escherichia coli (57.0%-57.4%), Klebsiella pneumoniae (7.5%-8.3%), Enterococcus faecium (6.8%-8.7%), Enterococcus faecalis (5.5%-6.0%) and Proteus mirabilis (3.3%-3.5%). Resis-tance rates of Enterococcus faecalis from urine specimens of male and female patients to ampicillin and nitrofurantoin were <12% and 7% respectively, to vancomycin was <3%; resistance rates of Enterococcus faecium to ampicillin and levofloxacin were both about 90%, to vancomycin were <4%. Resistance rates of Escherichia coli to ceftria-xone were>47%, to cefoperazone/sulbactam, piperacillin/tazobactam, and nitrofurantoin were all ≤8%, resis-tance rate to β-lactams in male patients was higher than in female patients, resistance rate to ceftriaxone was about 12 percentage higher. Resistance rates of Klebsiella pneumoniae isolated from male and female patients to ceftria-xone were about 58% and about 45% respectively. Resistance rates of Pseudomonas aeruginosa isolated from urine specimens of male and female patients to cefoperazone/sulbactam and piperacillin/tazobactam were both <14%, resistance rates to carbapenems were about 15%. Resistance rates of Acinetobacter baumannii to cefoperazone/sulbactam and minocycline were <27% and <22% respectively, resistance rates to carbapenems were 31.7%-47.7% in males and 26.5%-41.2% in females. Conclusion Constituent of bacteria isolated from urine specimens is different in different genders, antimicrobial resistance rates of partial Enterobacteriaceae are different among different genders, resistance rates of partial bacteria also changed in different years. Surveillance on antimicrobial resistance of bacteria isolated from urine specimens can provide reference data for the rational use of antimicrobial agents in urinary tract infection.