Abstract: An experiment was conducted to study the effect of epidermal growth factor(EGF),glutamine(Gln) and porcine growth release factor gene plasmid(pGRF gene plasmid)on the development of the digestive system in weaned piglets. In this trial, thirty piglets(Landrace × Large white × Taihu, initial weight 4.51 kg),weaned at 28 days, were randomly allocated to five treatment, three replicates of two piglets each. Pigs in different group were given different combination of the substances metioned above. Group I was given EGF alone; Group Ⅱ was given EGF and Gln; Group Ⅲ was given EGF and pGRF gene plasmid; Group Ⅳ was given EGF, Gln and pGRF gene plasmid; Group Ⅴ was given physiological saline as control. Samples were collected 21 days later. The weight of main digestive organs including stomach, liver, pancreas, small intestine and mucous of intestine were measured. The length of villi, the value of ALP in mucous of small intestine and the value of bacterial shift were detected. The results showed that the effect of the combined of EGF, Gln and pGRF gene plasmid was much better than that of EGF. Compared with control group, the weight of small intestine, frontal jejunum, middle jejunum, rear jejunum and entire jejunum of group Ⅳ were inereased by 37.12%(P<0.1),103%(P<0.05),8627%(P<0.05),100%(P<0.1) and 96.02%(P<0.05) respectively. In group Ⅳ, the length of villi in jejunum was increased by 89.68% and no bacterial translation was found. But the activity of alkaline phosphate enzyme wasn’t influnced significantly in every treated groups （P＞0.05）.