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Multi-drug resistant (MDR) Gram-negative bacteria have become one of the problems of nosocomial infection in China.The in-vitro antimicrobial activities of biapenem, arbekacin, cefminox and the other thirteen comparators against every non-repeat 100 E.coli, K.pneumoniae, A.baumannii, P.aeruginosa and 99 K.pneumoniae collected from six teaching hospitals across China in 2012 were evaluated by the broth microdilution method.The carbapenems remained high antimicrobial activities against E.coli (98%) and K.pneumoniae (more than 95%), followed by arbekacin and amikacin.The susceptibility rate of E.coli to cefminox was 96%.The MIC50 of E.coli to imipenem were 2 fold higher than biapenem and meropenem.MIC50 of E.coli to arbekacin were 4 fold lower than amikacin.The MIC50 and MIC90 of K.pneumoniae to meropenem was 4 and 8 fold lower than both of biapenem and imipenem, respectively.Both of the MIC50 and MIC90 of K.pneumoniae to arbekacin were 2 fold lower than amikacin.The resistance rates of A.baumannii to all antibiotics were more than 50% except for colistin.Amikacin and arbekacin were the most active agents against most of the P.aeruginosa isolates, followed by colistin (93%).The MIC50 of P.aeruginosa to arbekacin was 2 fold lower than amikacin.The MIC50 of P.aeruginosa to biapenem were 2 fold lower than imipenem and meropenem.In conclusion, biapenem and arbekacin remained high antimicrobial activities against Gram-negative bacteria except for A.baumannii, and cefminox maintained high antimicrobial activities against E.coli.