Objective: To assess the clinical value of double contrast-enhanced ultrasonography (DCEUS) of the stomach combined with determination of serum macrophage inflammatory protein 1 (MIP-1) and vascular cell adhesion molecule 1 (VCAM-1) in preoperative staging for gastric cancer. Methods: Six-hundred and eighty-five patients with gastric cancer underwent gastroscopy and DCEUS examination for preoperative staging, and meanwhile, their preoperative MIP-1 and VCAM-1 levels were measured by ELISA assay. According to the results of postoperative pathological tumor staging, the accuracy in preoperative stage estimation of gastric carcinoma between DCEUS and DCEUS plus MIP-1 and VCAM-1 measurement was compared. Results: The sensitivity (specificity) of DCEUS in estimating T stage of gastric cancer was 93.10% (92.05%) for T1, 67.47% (65.50%) for T2, 78.62% (80.47%) for T3 and 91.41% (90.70%) T4 respectively, with an overall accuracy rate of 80.15%, in estimating N stage was 90.55% (80.99%) for N0, 63.57 % (73.87%) for N1, 88.40% (92.50%) for N2 and 82.35% (73.68%) for N3 respectively, with an overall accuracy rate of 82.92%, and in estimating M stage was 99.29% (84.82%) for M0 and 71.48% (98.43%) for M1 respectively, with an overall accuracy rate of 88.61%. Both serum levels of MIP-1 and VCAM-1 were significantly related to the degree of invasion, lymphatic metastasis, distant metastasis and pathological stage of gastric carcinoma (all P<0.05). The sensitivity (specificity) of DCEUS plus MIP-1 and VCAM-1 measurement in estimating T stage of gastric cancer was 93.10% (92.05%) for T1, 87.95% (94.19%) for T2, 95.07% (92.33%) for T3 and 91.41% (90.70%) for T4 respectively, with an overall accuracy rate of 92.41%, in estimating N stage was 98.43% (96.90%) for N0, 89.15% (94.26%) for N1, 95.22 % (95.22%) for N2 and 92.65% (89.36%) for N3 respectively, with an overall accuracy rate of 94.16%, and in estimating M stage was 99.76% (97.68%) for M0 and 96.20%(99.61%) for M1 respectively, with an overall accuracy rate of 98.39%. The overall accuracy rates of DCEUS plus MIP-1 and VCAM-1 measurement in estimating either T, N or M stage of gastric cancer was significantly higher than that of DCEUS alone (all P<0.05). Conclusion: Serum levels of MIP-1 and VCAM-1 are closely related to pathological stage of gastric cancer, and DCEUS in combination with MIP-1 and VCAM-1 detection may help increase the accuracy rate of preoperative staging of gastric cancer.