To provide reference for rational use of antibiotics in clinic, this study investigated the distribution and antimicrobial resistance of pathogens causing bloodstream infection. The main pathogenic bacteria isolated from positive blood culture specimens and their antimicrobial resistance in Shanghai East Hospital from 2014 to 2021 were statistically analyzed. After eliminating the duplicate strains, a total of 1 396 strains of pathogenic bacteria were detected in blood culture samples during the past eight years. The gram-negative bacteria, gram-positive bacteria and fungi account for 56.3%, 38.4% and 5.3%, respectively. The top eight strains were Escherichia coli (21.5%), Coagulase negative Staphylococcus (CNS) (17.4%), Klebsiella pneumoniae (14.9%), Staphylococcus aureus (7.2%), Acinetobacter baumannii (5.4%), Enterococcus faecalis (4.2%), Enterococcus faecium (3.7%) and Pseudomonas aeruginosa (3.5%). The resistance rate of Escherichia coli to third-generation cephalosporins exceeded 50%, and the resistance rate to carbapenems was as low as 1%; The resistance rate of Klebsiella pneumoniae to carbapenems was 46.3%; The cumulative resistance rate of Acinetobacter baumannii to cephalosporins, aminoglycosides and carbapenems was more than 45%; The cumulative rates of methicillin resistant Staphylococcus aureus (MRSA) and methicillin-resistant coagulase-negative Staphylococci (MRCNS) were 44.6% and 74.5%, respectively.