Objective: A questionnaire survey was conducted on the implementation effect of the policy of “public welfare first-class guarantee and public welfare second-class management” in public health institutions in Wuhan, so as to provide references for the government to deepen the reform of the public health system. Methods: A cross-sectional survey and cluster sampling methods were employed, by using the self-designed questionnaire to survey data of the target population. The questionnaire results were statistically analyzed using Excel 2007 and SPSS 22.0. Results: The mean understanding of the “public welfare first-class guarantee and public welfare second-class management” policy among public health institution staff was 2.69±0.71, and there were statistically significant differences in population distribution (P＜0.05). The policy was found to promote the development strategy of the institution and it had a promoting effect to implement institution public welfare function. Policy implementation had a positive impact on staff career development and could stimulate staff work enthusiasm. Public health institutions generally had low levels of satisfaction with the current salary levels and performance assessment mechanisms of their units. Conclusion: It is necessary to strengthen the policy reform of “public welfare first-class guarantee, public welfare second-class management”. The pilot effects of “public welfare first-class guarantee, public welfare second-class management” policy has a rising space. In order to enhance confidence in the future development of public health institutions, it is necessary to give policy support in many aspects such as price level for approval, the number of establishment staff and financial investment guarantee mechanism.