Objective To explore the effect of nursing subspecialty construction in clinical departments. Methods The pilot program aiming at building nursing subspecialties in clinical departments was launched, with the department of neurology serving as the representative pilot unit. To make comparison, the program was not rolled out in some departments, with the department of respiratory diseases serving as the representative non-pilot unit.The pilot unit at department level built up a nursing subspecialties construction team, which, on basis of clinical subspecialty and physician-nurse partnership, developed specialty-oriented nursing care and core technology to constantly enrich the connotation of nursing service, and to promote more specialization and refinement of nursing. Results The pilot unit achieved higher rankings of scoring rate in nursing specialty construction, and achieved higher scores in nursing practice environment, than the non-pilot unit (P＜0.05,P＜0.01). Conclusion Nursing subspecialty construction can enrich the connotation of nursing specialties, accelerate talents cultivation, create a healthy nursing practice environment, and improve patient experience of health seeking.