Objective To describe the occurrence of neonatal procedural pain in NICU, and to provide basic data and references for neonatal pain management. Methods Totally, a convenient sample of 120 neonates admitted to NICU were recruited.A demographic and disease information questionnaire and an occurrence of procedural pain questionnaire were designed and relevant data were collected. A designated researcher observed at bedside the occurrence of neonatal procedural pain which were resulted by diagnostic, therapeutic, and nursing procedures.The intensity of pain was mea-sured by using the Neonatal Infant Pain Scale （NIPS）. Results The neonates experienced a total of 8 200 painful procedures.The neonates were exposed to a median of 20（interquartile rage:7,58） painful procedures per baby during hospitalization and a median of 5（2,10） painful procedures per baby per day.Respiratory system-related procedures were performed the most frequently （3 170 counts,38.7%）,and auxiliary diagnostic examinations were performed the least frequently （20 counts,0.2%）. All the 23 procedures listed in the questionnaire were painful, and 82.6%（19/23） of the procedures caused severe pain or more. Retinopathy of prematurity （ROP） screening and peripherally inserted central catheter （PICC） were the most painful procedures （pain score:7.0）, and venous injection was the least painful procedure（pain score:2.5±1.5）. Conclusion Painful procedures with high pain intensity were performed frequently in NICU neonates, which is in need of medical workers′ attention. Measures should be taken to alleviate neonatal procedural pain.