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The increasing prevalence of antimicrobial resistance (AMR) infections in children is becoming a growing global concern. AMR poses a significant challenge in pediatric diarrhea, where multidrug-resistant Enterobacteriaceae threaten treatment efficacy. This study investigates the prevalence, resistance mechanisms, and genetic characteristics of carbapenem-, tigecycline-, and polymyxin-resistant Enterobacteriaceae isolated from pediatric diarrhea cases , in 2017 (n=1059) and 2023 (n=367). The prevalence of mcr-1-positive strains declined significantly from 2.36% (25/1059) in 2017 to 0.82% (3/367) in 2023, while tet(X4)-positive strains rose from 0% to 1.91% (7/367). Carbapenemase-producers remained rare (0.28% in 2017, 1.63% in 2023), with blaNDM as the dominant carbapenemase gene. Whole genome sequencing revealed diverse antimicrobial resistance genes (ARGs) and evolving plasmid types. Notably, by 2023 all mcr-1 were carried on IncI2 plasmids. Conjugation experiments confirmed plasmids carrying mcr-1, blaNDM-5, and tet(X4) are transferable to a recipient strain, highlighting the potential for horizontal gene transfer. Our findings demonstrate a significant shift in pediatric diarrhea-associated Enterobacteriaceae resistance profiles from 2017 to 2023. The decline in mcr-1 mirrors the impact of reduced polymyxin use, whereas the rise of tet(X4) signals a new challenge. These findings underscore the dynamic nature of AMR in pediatric Enterobacteriaceae and emphasize the need for stringent antimicrobial stewardship and surveillance. A One Health approach, integrating clinical, agricultural, and environmental efforts, is crucial to mitigating AMR spread and protecting pediatric health globally.