Objective??To analyze distribution and antibiotic resistance pathogenic bacteria in neonatal sepsis in 10 years. Methods The distribution of pathogens and their antibiotic resistance were retrospectively analyzed in neonatal sepsis from January, 2004 to December, 2013. The results were compared between 2004-2008 and 2009-2013. Results The percentage of Streptococcus agalactiae rose from 1.0% to 4.2% and fungi rose from 1.9% to 7.9% in all pathogens in past 10 years. But the distribution of pathogenic bacteria was not significantly different. The ESBLs of Escherichia coli were increased from 28.6% to 36.0% (P>0.05). The Escherichia coli resistant to imipenem and meropenem had not been found. The Staphylococcus resistance to oxacallin and ampicillin/sulbactam was increased. The Staphylococcus resistant to vancomycin had not been found. Conclusions The main pathogens of neonatal sepsis are coagulase-negative staphylococci and E. coli. The fungi and Streptococcus agalactiae infections are significantly increased.