Abstract: 【Abstract】 Objective To study the use of antibiotics in neonatal intensive care unit (NICU) and the prognosis of the newborns using strategy of strictly controlling antibiotic use. Methods A retrospective analysis was performed on the medical data of 5 162 neonates discharged from January 2016 to June 2018.According to the strategy of antibiotic use, they were divided into two groups: preliminary control group (n=2 065) and strict control group (n =3 097). The rate of antibiotic use, the types of antibiotics, the mortality and morbidity of complications of the newborns, and the strategy of antibiotic use were analyzed. Statistical methods included t-test of two independent samples, χ2 test and Fisher's exact test. Results ① Among 5162 newborns, 2 986 (57.85%) were males, with an average gestational age of (38.2±2.3) weeks and an average birth weight of (3 027±586) g and there were 1 135 (21.99%) critical ill newborns. ② Among 5 162 newborns, 1 225(23.73%) newborns had used antibiotics. The top five antibiotics used were ampicillin, piperacillin tazobactam, penicillin, erythromycin and cefotaxime sodium sulbactam sodium. Third-line drugs were rare. Nine neonates were treated combined with three types of antibiotics, 145 neonates combined with two types of antibiotics, and the rest with only one type of antibiotic. ③ The antibiotic use rate of neonatal pneumonia was 61.18% (446/729), of which Chlamydia trachomatis pneumonia and Ureaplasma urealyticum pneumonia were 100.0%, using oral erythromycin dry suspension or azithromycin dry suspension. The antibiotic use rates of newborns diagnosed with meconium contamination of amniotic fluid, newborns withmaternal premature rupture of membranes, neonatal asphyxia, pathological jaundice (the first diagnosis),necrotizing enterocolitis (NEC) and suspected NEC, and preterm infants were 30.12% (153/508), 35.73%(144/403), 44.53% (118/265), 6.35% (89/1402), 78.02% (71/91), 40.39% (374/926). ④ Prophylactic use ofantibiotics in strict control group was significantly lower than that in preliminary control group (9.81% vs15.59%), while the incidence of NEC and suspected NEC was lower than that in preliminary control group(1.36% vs 2.37%, P<0.05). There was no significant difference in the mortality rate, the incidence of fungal infection and the incidence of nosocomial infection between the two groups. Conclusions Following the strict antibiotic use strategy can reduce the unnecessary antibiotic use, especially the preventive application and the application of third-line antibiotics, and reduce the complications of neonates.