Abstract: Objective To evaluate the application of dynamic changes in amplitude-integrated electroencephalography (aEEG), serum neuron-specific enolase (NSE), and plasma neuropeptide Y (NPY) in the diagnosis and treatment of neonatal hyperbilirubinemia. Methods A total of 156 full-term newborns with hyperbilirubinemia who were admitted to the neonatal intensive care unit of Xuzhou Central Hospital from January to December 2022 were selected. According to the bilirubin-induced neurologic dysfunction (BIND) assessment scale scores, the patients were divided into two groups: a control group (n=108, with no brain injury) and a study group (n=48, with brain injury). Both groups underwent dynamic aEEG monitoring. Both groups were compared for their levels of serum NSE and plasma NPY and aEEG background activity, in order to assess the predictive value of serum NSE, plasma NPY, and aEEG alone or in combination, for early detection of brain injury due to hyperbilirubinemia in newborns. Results The serum NSE and plasma NPY levels in the study group were significantly higher than those in the control group, with statistical differences (P<0.001). Both NSE and NPY levels were positively correlated with the occurrence of brain injury in newborns with hyperbilirubinemia. Abnormal aEEG background activity was also positively correlated with the occurrence of brain injury in newborns with hyperbilirubinemia, with significant differences (P<0.05). The area under the curve (AUC) of early prediction of brain injury in newborns with hyperbilirubinemia was 0.981 for NSE levels, 0.877 for NPY levels, and 0.853 for aEEG background activity; the AUC for the combined prediction was 0.987. Conclusions The combination of dynamic aEEG monitoring and measurements of serum NSE and plasma NPY levels provides better early prediction of brain injury in newborns with hyperbilirubinemia, which is worthy of clinical promotion.