Squamous cell/adenosquamous carcinoma (SC/ASC) of the gallbladder are rare tumors and there arefew clinical reports in the literature. Herein we report our clinical experience with 46 patients with SC/ASCand 80 with adenocarcinoma (AC). Expression of EphB1 and Ephrin-B in each tumor was determined usingimmunohistochemical methods for determination of correlations with prognosis. There was no difference in EphB1and Ephrin-B expression between SC/ASC and AC tumors (P>0.05), but greater expression in those less than3 cm in diameter, stage Ⅰ or Ⅱ (TNM stage), with no lymph node metastases, with no local invasion and treatedwith radical resection was apparent. Expression of EphB1 (P<0.05) and Ephrin-B (P<0.01) was higher in welldifferentiated than in poorly differentiated AC tumors. Kaplan-Meier survival analysis indicated that degreeof differentiation, tumor diameter, lymph node metastases, local invasion, surgical approach and expressionrate of EphB1 and Ephrin-B were closely related to the survival of SC/ASC (P<0.05) and AC patients (P<0.01).Patients with tumors that positive expressed EphB1 and Ephrin-B, whether it is SC/ASC (P SC/ASC =0.000) orAC (P AC =0.000 or P AC =0.002) had longer survival than those negative expression. Cox multivariate analysisindicated a negative correlation between expression of EphB1 or Ephrin-B and overall survival. Hence, EphB1and Ephrin-B could be regarded as independent good prognostic factorsand important biological markers forSC/ASC and AC of gallbladder.