Objective: To understand physical fitness among primary and middle school students in Henan Province, so as to provide theoretical basis for physical fitness promotion among primary and middle school students. Methods: Based on data of National Survey on Student Physical Fitness and Health in Henan Province in 2019, a total of 42 734 students were investigated. The Z value of each physical fitness index were calculated by Z method, physical fitness index (PFI) was obtained to reflect the physical fitness condition synthetically. Physical fitness of different groups were compared, and associated factors of PFI were analyzed by multiple linear regression. Results: Lung capacity, grip strength, standing long jump, 50 m run, 50 m×8 shuttle run and PFI in boys were [(2 552.1±1 226.5)mL, (26.0±13.9)N, (171.0±43.7)cm, (9.3±1.8)s, (132.1±22.6)s, (1.4±3.6)], respectively, which were significantly higher than that of girls [(1 965.2±765.3)mL, (19.1±8.2)N, (142.4±26.6)cm, (10.5±1.4)s, (136.9±21.1)s, (-1.4±3.2)]( t =59.35,62.66,81.87,-74.92,-16.72,85.96, P <0.01). The seated forward flexion of boys [(7.0±7.5)cm] was significantly lower than that of girls[(11.8±7.1)cm], the difference was statistically significant ( t =-68.57， P <0.01). Significant sex differences were observed in physical fitness at different age ( P <0.01). There was a certain gap between urban boys and rural boys in terms of strength quality, endurance quality and flexibility quality, and the physical quality of urban girls was better than that of rural girls as a whole. The detection rates of poor eyesight, malnutrition, overweight and obesity, anemia and high blood pressure of primary and middle school students were 68.0%, 5.4%, 26.0%, 15.9% and 18.0%, respectively. Region, sex, age and nutritional status(malnutrition,overweight and obesity) were the influencing factors of PFI( β =-0.23,-2.92,-0.11,-1.19, -0.78 , P <0.05). Conclusion Physical fitness among primary and middle school students varies in gender, age group, region and disease condition in Henan Province. Scientific intervention should be administered, especially for rural, female students, senior students, undernourished and overweight and obese students, to improve health awareness and physical fitness.