In IHC, staining percentage scores (0-5 points) multiplied staining intense scores (0-5 points) to get IHC scores (0-25 points). Staining percentage scores were defined as 0 point (0%-5%), 1 point (6%-25%), 2 points (26%-50%), 3 points (51%-75%), 4 points (76%-95%) and 5 points (96%-100%). Staining intense scores were defined as 0 point (negative), 1 point (weak), 2 points (weak to moderate), 3 points (moderate), 4 points (moderate to strong) and 5 points (strong). With the use of X-tile software (Version 3.6.1, Yale University), the optimal cut-points for IHC were analyzed and calculated as 12 points of CD54, 0 point of Lgr5, 0 point of CD133, 0 point of Oct4, 6 points of EpCAM and 8 points of Sox2 in primary lesions. The cut-points of these markers in metastatic LNs were the same with primary lesions, except Lgr5 (5 points) and Oct4 (1 point). Regarding qPCR, the cut-points of the expression folds of GC tissues to corresponding adjacent normal gastric tissues were also analyzed and calculated through X-tile. The cut-points were 1.5 folds of CD54, 1.3 folds of Lgr5, 2.5 folds of CD133, 0.6 folds of Oct4, 7 folds of EpCAM and 0.2 folds of Sox2. According to the cut-points in IHC and qPCR, the patients were further divided into low expression groups (≤cut-points) and high expression groups (>cut-points).