BACKGROUND: Assessment of physical fitness is a common and effective method for evaluating the health status of teenage students. OBJECTIVES: The present study was conducted to assess physical fitness in adolescents residing in Shanghai, and investigate the relationships among anthropometry, muscle fitness, and a 20-meter shuttle run test (20-m SRT). METHODS: A total of 449 middle school students (12–17 years old, 246 boys and 203 girls) from four different regions of Shanghai, China were included as study participants. RESULTS: Multiple linear regression analyses indicate that upper extremity muscle strength and standing vertical jump were positively related to age and weight, while negatively related to gender, scapular skinfold, and calf skinfold. Strong positive correlations were also demonstrated between the number of laps accomplished on the 20-m SRT and upper extremity muscle strength as well as standing vertical jump height. CONCLUSION: Muscle strength is an important anthropometric characteristic that may be used for the assessment of sprinting ability in adolescents.