Objective: To explore the relationship between miR-181b rs322931 polymorphism and the risk of gastric cancer. Methods: The peripheral venous blood of 172 patients with gastric cancer and 224 healthy controls were collected. The miR-181b rs322931 was typed by TaqMan probe method, and correlation between miR-181b rs322931 and the risk of gastric cancer was evaluated using SPSS software. Results: The frequencies of miR-181b rs322931 CC, CT, TT genotype, C, and T allele were 61.6, 33.7, 4.7, 78.5, and 21.5% in gastric cancer, and 74.1, 23.7, 2.2, 85.9, and 14.1% in controls, respectively. After the ?2 test and correction for age and gender, the risk of gastric cancer in carriers of CT and CT/TT genotypes increased by 1.71 and 1.79 times, respectively (CT vs. CC: 95% CI, 1.10-2.67, P=0.02; CT /TT compared with CC: 95%CI, 1.16-2.74, P=0.008). Moreover, compared with alleles, the risk of gastric cancer in T allele carriers increased by 1.69 times (95%CI, 1.16-2.44, P=0.006). Conclusion: miR-181b rs322931 polymorphism may be one of the susceptibility genes of gastric cancer in the Chinese Han nationality.