Objective To investigate head nurses′ knowledge and management of in-hospital standardized insulin injections, and to analyze the influencing factors. Methods By using stratified sampling method, a total of 796 head nurses from 25 hospitals in Hubei Province were selected. The general information questionnaire, the Insulin Injection Knowledge Questionnaire, and the In-hospital Homogeneous Administration of Insulin Therapy Questionnaire were used to survey the head nurses. Results The head nurses scored (14.10±3.26) points in insulin injection knowledge and (11.48±2.81) points in in-hospital homogeneous administration of insulin therapy. Department and education levels were the main factors influencing head nurses′ scores in insulin injection knowledge （P<0.05 for both）. Hospital levels and knowledge scores were the main factors influencing head nurses′ scores in hospital homogeneous administration of insulin therapy （P<0.05 for both）. Conclusion Head nurses′ knowledge of in-hospital standardized insulin injections is low, while their in-hospital homogeneous administration of insulin therapy is at medium level. Further training is needed to enhance standardization of insulin injection management across the hospital.