There are some published studies focus on the invert U-shaped relationship between fasting plasma glucose (FPG) and serum uric acid (UA), while the threshold value and gender differences of this relationship were still obscure. We aimed to explore the dose-response relation between FPG level and serum UA concentration by conducted this epidemiological research in a large health check-up population in China. A total of 237,703 people were collected from January 2011 to July 2014 in our cross-sectional study; 100,348 subjects age 18 to 89 years and without known diabetes were included for the current analysis. One-way analysis of variance, generalized additive models, and 2-piecewise linear regression model were used. The mean concentration of UA with FPG of <6.1, 6.1 to 6.9, and ≥7.0 mmol/L was 240.9, 260.2, and 259.6 μmol/L in women and 349.0, 360.8, and 331.0 μmol/L in men. An invert U-shape with a threshold FPG of 7.5 (women)/6.5 (men) mmol/L was observed in the regression curve of FPG and UA, even after adjusting for potential confounders. The adjusted regression coefficients were 2.4 (95% confidence interval [CI]: 1.5 to 3.4, P < 0.001) for FPG < 7.5 mmol/L, -3.2 (95% CI: -5.0 to -1.3, P < 0.001) for FPG ≥ 7.5 mmol/L in women; while 0.8 (95% CI: -0.4 to 2.0, P = 0.19) for FPG < 6.5 mmol/L, -7.1 (95% CI: -8.0 to -6.1, P < 0.001) for FPG ≥ 6.5 mmol/L in men. Furthermore, the interaction between different FPG level and sex was significant (P < 0.05). An invert U-shape with a threshold of FPG was existed for serum UA level in Chinese adults age 18 to 89 years without known diabetes, and significant gender differences were found. Future researches should pay more attention to this relationship.