Objective To understand the distribution and antimicrobial resistance of bacteria isolated from patients in department of urology in China in 2021. Methods According to the technical program of China Antimicrobial Resistance Surveillance System (CARSS), data and antimicrobial susceptibility testing results of bacteria isolated from patients in department of urology reported by all CARSS member units in 2021were analyzed with WHONET 5.6 software. Results A total of 232 603 strains of bacteria were isolated from urological patients, 166 483 (71.6%) were Gram-negative bacteria and 66 120 (28.4%) were Gram-positive bacteria. The top 5 specimens were urine, blood, wound secretion, sputum, and abdominal peritoneal effusion. The top 5 isolated Gram-negative bacteria were Escherichia coli(57.5%), Klebsiella pneumoniae(10.8%), Pseudomonas aeruginosa(5.8%), Proteus mirabilis(5.3%), and Enterobacter cloacae(3.4%). The top 5 isolated Gram-positive bacteria were Enterococcus faecalis(36.2%), Enterococcus faecium(14.9%), Staphylococcus epidermidis(10.5%), Streptococcus lactis(9.5%), and Staphylococcus aureus(7.3%). Vancomycin-, teicoplanin-, and linezolid-resistant Staphylococcus aureus was not found. The resistance rates of methicillin-resistant Staphylococcus aureus to gentamicin, rifampicin, levofloxacin, cotrimoxazole, clindamycin, and erythromycin were all higher than those of methicillin-sensitive Staphylococcus aureus. The resistance rates of Escherichia coli, Klebsiella pneumoniae, and Proteus mirabilis to imipenem(1.1%-5.3%), meropenem(0.7%-5.9%), and ertapenem (0.7%-4.4%) were relatively low, and the resistance rates to ceftriaxone ranged from 36.5% to 54.5%. Resistance rates of Enterobacterales to tegacyclin, polymyxin B, and ceftazidime/avibactam were relatively low. Resistance rates of Pseudomonas aeruginosa and Acinetobacter baumannii to imipenem, meropenem, polymyxin B, tegacyclin, ceftazidime/avibactam, and enzyme inhibitor agents were relatively low. Conclusion The major specimen from department of urology is urine, the main isolated pathogenic bacteria are Escherichia coli and Klebsiella pneumoniae, and antimicrobial resistance rate of Enterobacterales is different. Surveillance on the antimicrobial resistance of bacteria isolated from urine specimens can provide reference for the rational use of antimicrobial agents in urinary tract infection.