Background <p>With the rapid development of non-public tertiary general hospitals in China, the construction of patient safety culture has become a priority to narrow the gap with public hospitals in terms of patient safety management, however, the current driving factors have not been identified. This study aimed to evaluate the current status of patient safety culture in a non-public tertiary general hospital and to identify the principal influencing factors. In addition, the study sought to verify the construct validity of the Hospital Survey on Patient Safety Culture (HSOPSC) scale adapted for samples from non-public tertiary hospitals.</p> Methods <p>A cross-sectional survey on patient safety culture was conducted among 840 medical staff members in a non-public tertiary general hospital from June to August 2025 using the Chinese version of the HSOPSC scale, which comprises six dimensions and 39 items. Stratified cluster sampling was adopted, with stratification first performed according to clinical departments, followed by cluster sampling, with each department treated as a sampling cluster. Item screening and scale optimization were conducted through item discrimination analysis, factor loading evaluation, and reliability assessment. One-way analysis of variance was applied for the preliminary comparison of the total mean patient safety culture scores across medical staff with different characteristics. Statistically significant variables from the one-way analysis were included in a multivariate logistic regression model to identify independent factors affecting the perception of patient safety culture. Department type, working tenure, and professional category were taken as the core independent variables, and their impacts on the perception of team patient safety culture were evaluated.</p> Results <p>Following adjustment and refinement, the revised HSOPSC scale retained 29 items and demonstrated an overall Cronbach’s α coefficient of 0.95. The overall positive response rate for “Patient Safety Culture” was 66.65%. “Managerial Support for Patient Safety” (77.70%), “Colleague Collaboration” (76.15%), and “Communication” (75.43%) were strengths, whereas “Staff Recognition of Hospital Safety Culture” (58.21%) and “Incident Reporting” (66.47%) were weaknesses. Multivariate analysis identified department as an independent influencing factor. Medical staff in critical care (OR = 0.42), laboratory (OR = 0.25), and platform departments (OR = 0.49) had significantly lower safety culture perception (P &lt; 0.05).</p> Conclusion <p>Patient safety culture in the non-public tertiary general hospital is at a moderate level, with marked departmental differences. Future interventions should prioritize fostering a non-punitive reporting environment, improving staffing in high-risk units, and strengthening interdepartmental communication and collaboration to enhance the overall patient safety culture climate.</p>