Objective: To evaluate the clinicopathological characteristics of gastric cancer patients with perioperative serum carbohydrate antigen 19-9 (CA19-9)-positive and alpha-fetoprotein (AFP)-positive, and combined effect of perioperative serum CA19-9 and AFP on clinicopathological characteristics of gastric cancer patients. Methods: The clinicopathological characteristics of 900 gastric cancer patients with serum CA19-9 and AFP detection perioperatively from the First Affiliated Hospital of NJMU were analyzed retrospectively. Results: Patients with elevated serum CA19-9 had the characteristics of more advanced T stage (T4 stage, 94.2% vs. 65.5%), later clinical stage (Ⅲ＋Ⅳstage ,89.2% vs. 61.5%), more common lymph node metastasis (81.5% vs. 59.1%) and more remarkable vascular as well as neural invasion compared with those with normal level of serum CA19-9 (32.5% vs. 23.3%; 42.5% vs. 27.1%); Compared with AFP-negative patients, the AFP-positive patients suffered later clinical stage (Ⅲ＋Ⅳstage ,88.9% vs. 60.6%), more common lymph node metastasis (81.5% vs. 59.1%) and more marked neural invasion (44.4% vs. 27.0%) (P＜0.05). Patients with one or two positive tumor markers suffered more advanced T stage, later clinical stage, more common lymph node metastasis and more remarkable vascular as well as neural invasion compared with those with double-negative tumor markers (P＜0.05). Two-category logistic regression analysis showed that CA19-9(+) was the independent risk factor for GC T stage; compared with CA19-9(-)-AFP(-), CA19-9(+)-AFP(+) was the independent risk factor for GC vascular invasion, CA19-9(+)-AFP(-) was the independent risk factor for GC T stage (P＜0.05). Conclusion: Perioperative serum levels of CA19-9 and AFP are closely correlated with clinicopathological features for patients with gastric cancer, and the two tumor markers are highly recommended detecting perioperatively for offering information of clinicopathological characteristics.