Purpose: The purpose of this study was to examine the prevalence of four important drug-resistance phenotypes: difficult-to-treat resistance (DTR), fluoroquinolone resistance (FQR), carbapenem resistance (CR), and extended-spectrum cephalosporin resistance (ECR). Methods: DTR was defined as insensitivity to all the beta-lactams and fluoroquinolones tested. We retrospectively analyzed the distribution characteristics of specific drug-resistant phenotypes of the main Gram-negative bacteria causing bloodstream infections (BSIs) in Tongji Hospital (Wuhan, China) between 2013 and 2021: Escherichia coli, Klebsiella pneumoniae, Pseudomonas aerugi-nosa, and Acinetobacter baumannii. Results: FQR was the main antimicrobial resistance phenotype of E. coli, accounting for 59.45% (1117/1879, 95% confidence interval, 57.21%-61.65%); the detection rates for CR and DTR were low, accounting for 1.86% (35/1879, 1.34%-2.58%), and 1.81% (34/1879, 1.30%-2.52%), respectively. However, the detection rates for CR and DTR in K. pneumoniae were 38.83% (497/1280, 36.20%-41.53%) and 35.94% (460/1280, 33.35%-38.60%), respectively. In P. aeruginosa, the detection rates of the four drug-resistant phenotypes (DTR, CR, FQR, and ECR) were all < 30%, but conversely, for A. baumannii, the detection rates were all > 80%. The changes in the data from 2013 to 2021 showed upward trends (z > 0) for CR-E. coli, DTR-E. coli, FQR-E. coli, CR-K. pneumoniae, DTR-K. pneumoniae, FQR-K. pneumoniae, and ECR-K. pneumoniae, but downward trends (z < 0) for ECR-E. coli, CR-A. baumannii, DTR-A. baumannii, FQR-A. baumannii, ECR-A. baumannii, CR-P. aeruginosa, DTR-P. aeruginosa, FQR-P. aerugi-nosa, and ECR-P. aeruginosa. Conclusion: DTR warrants further attention, especially in in BSI-associated K. pneumoniae and A. baumannii, in which the detection rates were very high. Between 2013 and 2021 in this region, DTR-E. coli and CR-E. coli showed obvious upward trends, whereas DTR-P. aeruginosa and ECR-P. aeruginosa showed obvious downward trends.