Objective To analyze the willingness and actual behavior to influenza vaccination among healthcare workers (HCWs) in a tertiary general hospital, and provide a stratified intervention basis for improving the vaccination rate of HCWs. Methods In March 2025, a cross-sectional survey among HCWs was conducted through the "Questionnaire Star". The survey participants were divided into willing/unwilling group and vaccinated/unvaccinated group, with "willingness to vaccination" and "actual vaccination" as dependent variables. The influencing factors of different dimensions were analyzed by χ²-test and logistic regression. Results A total of 720 questionnaires were distributed, and 708 valid questionnaires were collected, with an effective rate of 98.3%. The vaccination willingness rate (80.1%) was higher than the actual vaccination rate (45.9%) (χ²=72.65, P<0.01). The willingness group scored higher in influenza knowledge ([76.9±9.3] points vs [67.8±12.5] points, P<0.001). Previous influenza vaccination history (OR=4.33, 95%CI: 2.85-6.56, P<0.001) and influenza vaccine high awareness level (OR=1.08, 95%CI: 1.06-1.10, P<0.001) were the promoting factors. The actual vaccination behavior was influenced by multiple factors. Older age (OR=1.04, 95%CI: 1.01-1.07, P=0.020), medical technology series (OR=3.94, 95%CI: 1.68-9.25, P=0.002), management personnel (OR=14.97, 95%CI: 1.16-193.76, P=0.038), personnel with senior professional title (OR=4.69, 95%CI: 1.42-15.55, P=0.011), key departments (OR=1.71, 95%CI: 1.08-2.72, P=0.023), previous vaccination history (OR=12.21, 95%CI: 7.97-18.70), and high score of awareness level (OR=1.02, 95%CI: 1.01-1.04) had higher vaccination rates. Conclusion The willingness of HCWs to influenza vaccination is mainly driven by their internal cognition and previous experience, while their actual vaccination behavior is further influenced by the combination of occupational structure characteristics and external environmental factors, resulting in a significant gap between willingness and behavior. Medical institutions should optimize their education strategies for improving the awareness of HCWs, take multiple measures to eliminate barriers to behavior transformation, so as to increase vaccination rate and reduce the risk of influenza transmission within hospitals.