Objective To investigate the current status of professional values among healthcare workers in Sichuan Province and explore the associated influencing factors, thereby providing evidence for enhancing professional values and improving hospital management. Methods A cross-sectional study was conducted between October 2022 and March 2023. Healthcare workers from 25 hospitals in Sichuan Province were recruited using convenience sampling. A general information questionnaire and the Professional Values Scale for Healthcare Workers were administered to assess the current status of professional values and explore the influencing factors. Results A total of 4502 questionnaires were distributed, with 4267 valid ones returned, resulting in an effective recovery rate of 94.8%. The total professional values score among the 4267 healthcare workers was 114.95±21.12. The dimensions ranked in descending order of their mean item scores were as follows: intrinsic values (3.89±0.67), altruistic values (3.85±0.72), social values (3.57±0.68), extrinsic values (3.38±0.79), and leisure values (3.12±0.75). Multiple linear regression analysis revealed that salary-workload match was the most significant factor influencing professional values. After controlling for other variables, the “very poor match” group scored 38.87 points lower on average than the “excellent match” group (P<0.001). Health status also had a significant impact, with the “very unhealthy” group scoring 26.67 points lower on average than the “very healthy” group (P<0.001). Additionally, nurses scored 3.88 points higher on average than physicians (P<0.001), and males scored 2.10 points lower on average than females (P=0.014). Conclusions The professional values of healthcare workers are characterized by “intrinsic motivation and altruistic orientation”. Salary-workload match and health status are the most critical influencing factors. Healthcare institutions should prioritize pay equity, employee health management, and career development pathways, while adopting differentiated human resource management strategies.