Objective To investigate the change in clinical distribution and antimicrobial resistance of Klebsiella pneumoniae(KP), and provide evidence for KP infection control and treatment. Methods 10 075 strains of KP isolated from a hospital from 2009 to 2018 were analyzed retrospectively, constituent of KP in different years, different departments and different infection sites were compared, changing trend of resistance of KP was analyzed. Results KP mainly isolated from sputum specimens, which was the main pathogen of liver abscess and the most isolated pathogen from department of neonatology. Isolation rate of KP increased from 8.0% in 2009 to 13.8% in 2018, showing an upward trend year by year (χ_trend²=389.9, P<0.001). Resistance rates of KP to imipenem and meropenem increased from 0.6% and 1.5% to 34.4% and 32.8% respectively during 10 years; isolation rates of ESBLs-producing KP increased from 18.8% to 62.6%. KP isolated from general intensive care unit(ICU) and department of neurology showed the highest resistance rate to imipenem, rising from 1.9% and 0 to 63.7% and 52.9% respectively, antimicrobial resistance rates of KP from blood, sputum, urine, and abdominal drainage increased to from 0, 1%, 0, and 0 to 29.3%, 26.6%, 34.0%, and 22.9% respectively. Conclusion The isolation rate and antimicrobial resistance rate of KP showed a continuous upward trend during 10 years, isolation rate of CRKP also showed a continuous upward trend, there were differences among different departments and different infection sites.