Objective To explore whether the Chinese medicine Guben Yiliu Ⅲ can improve the effect of gemcitabine on human pancreatic cancer xenograft in nude mice. Methods Nude mice with transplanted human pancreatic cancer were divided randomly into 4 groups: control group, gemcitabine treatment group, combined (Guben Yiliu Ⅲ + gemcitabine) group, and Guben Yiliu Ⅲ group, 10 mice in each group. The gemcitabine group and combined group were treated with gemcitabine from the 8^th day after transplantation in a dose of 100 mg/kg by i.p. injection, twice a week. Guben Yiliu Ⅲ and combined groups were given the aqueous solution of Guben Yiliu Ⅲ granules p.o. since the 8^th day after transplantation. Result The inhibition rate of transplanted tumor in the three treatment groups were 48.9% in the gemcitabine group, 68.9% in the combined group, and 28.0% in the Guben Yiliu Ⅲ group. The combined group showed a significantly higher inhibition rate than the gemcitabine group (P<0.05). The gemcitabine group, combined group and Guben Yiliu Ⅲ group showed a significantly slower growth rate than the control group. However, the combined treatment group showed a pronounced side effect and body weight loss than the other 3 groups. Conclusions The Chinese medicine Guben Yiliu Ⅲ can improve the inhibitory effect of gemcitabine on nude mice with human pancreatic cancer xenograft in the auxilla of nude mice.