Objective: The critical values in health checkup play a key role in preventing chronic diseases and different types of cancer. This study aimed to analyze the prevalence, association relation, and dynamic evolution of critical values in health checkups at a large physical examination center in China.Methods: Herein, we chose 33,639 samples of physical examiners from January 2017 to December 2019. After strict exclusion processes, combined with the critical values in health checkup reporting data, 4,721 participants with at least one critical value were included. We first defined a critical value list for laboratory test, imaging, cervical cancer screening, electrocardiogram, and health checkup informed on site, and then performed a cross-sectional study to analyze the distribution and significance of critical values of 4,721 participants from different views and the association relation of 628 participants with more than one critical value and a retrospective cohort study to analyze the incidence and dynamic evolution of critical values based on 2,813 participants attending the physical examination from 2017 to 2019.Results: A total of 4,721 participants were included in the retrospective study. The prevalence of 10 critical values from 33,639 participants was over 0.6%. The critical values of obesity, hypertension, Glucose_T, Liver_T, Kidney_T, Lipid_T, Urine_T, and Head_CT were significantly increased in men (P < 0.05), whereas the results were the opposite for the Blood_T and Thyroid_US (P < 0.01). The prevalence trend of critical values increased along with age, where the prevalence of men was higher than that of women under 60 years old (P < 0.01), while the prevalence of women increased by four times and exceeded the prevalence of men above 70 years old. Association relation analysis identified 16 and 6 effective rules for men and women, respectively, where the critical values of Urine_T and Glucose_T played the central roles. Furthermore, a retrospective dynamic evolution analysis found that the incidence of new critical values was about 10%, the incidence of persistent critical values was about 50%, and that most of the effective evolution paths tended to no critical values for men and women.Conclusion: In conclusion, this study provides a new perspective to explore the population health status using the critical value reporting data in a physical examination center, which can assist in decision-making by health management at the population level and in the prevention and treatment of various types of cancer and chronic diseases at the individual level.