Abstract: Objective: To analyze the relationship between different gender, disease types, age of diagnosis, agglutination intensity of direct anti-human globulin test and degree of hemolysis and anemia in newborn children with hemolytic disease(HDN).Methods: A total of 104 cases of HDN newborn children who were diagnosed in our hospital from January 2016 to December 2019 were analyzed, and total bilirubin(TBIL), lactate dehydrogenase(LDH), hemoglobin(Hb) concentrations of all children were detected. According to gender, disease type, age of diagnosis and DAT agglutination intensity, all children were divided into different groups, and the differences in TBIL, LDH, Hb concentrations between the groups were compared and statistically analyzed.Results: Of the 104 HDN children, 48 cases were male, accounting for 46.15%, and 56 cases were female, accounting for 53.85%. There were no statistically significant differences in the concentrations of TBIL, LDH, and Hb among children of different genders(all P>0.05). Children with Rh-HDN had higher TBIL and LDH than that of ABO-HDN, and Hb concentration was lower than that of ABO-HDN, the differences were statistically significant(all P＜0.05). Among the ABO-HDN patients, the TBIL concentration in the O-A group was higher than that in the O-B group, and the Hb concentration was lower than that in the O-B group. The differences were statistically significant(both P＜0.05). The TBIL in the O-AB group was higher than that in the O-A group and O-B group, and the Hb was lower than that in the O-B group, the difference was statistically significant(both P＜0.05). Among Rh-HDN patients, TBIL and LDH in anti-D group were higher than those in anti--E/C/Ce group, and the differences were statistically significant(both P＜0.05). The TBIL concentration in the diagnosed day 4-7 days group was lower than that in the>7 day group, and the differences were statistically significant(both P＜0.05). TBIL and LDH concentrations in DAT4 + group were higher than those in 3+, 2+, ≤1 + group, Hb concentration was lower than that in 2+, ≤1 + group, TBIL, LDH concentration in DAT3 + group were higher than those in 2+, ≤1 + group, and the differences were statistically significant(all P＜0.05). Children with RH-HDN were likely to cause an increase in TBIL and LDH and a decrease in Hb. The age of diagnosis and DAT agglutination intensity were positively correlated with TBIL and LDH, and DAT agglutination intensity was negatively correlated with Hb(all P＜0.05).Conclusion: The gender of children with HDN had no correlaiton with the degree of hemolysis and anemia. For children with HDN caused by Rh, age of diagnosis>7 days and DAT agglutination intensity ≥3 +, it should pay close attention to changes in bilirubin and hemoglobin, and take timely measures to prevent bilirubin encephalopathy.