Objective To summarize the clinical application of esophagogastrostomy with layered anastomosis and to observe the healing quality of anastomotic stoma in animal experiments. Methods One thousand and twenty-four patients suffered from carcinoma of esophagus or carcinoma of gastric cardia had undergone esophagogastrostomy by layered anastomosis with absorbable suture. Twenty-four experimental dogs (adult male healthy hybrid dogs) were divided into two groups: the experimental group and the control group. The former (experimental group) underwent the layered anastomosis, the diameter of esophagogastric stoma and the length and depth of stomal scar were measured under anesthesia in both groups on 5th,8th,14th,and 42th postoperative day, respectively. So were done the histological measurement, such as the count infiltrating inflammatory cells, the proliferation of blood capillary and other cells. And the cytokines related to wound healing (LsAB technique) such as epidermal growth factor(EGF), transforming growth factor-beta 1 (TGF-beta 1) were detected, either. Results One thousand and twenty-four patients had no anastomotic leakage. There were only 6 patients suffered from mild anastomotic stricture, and they got well after one dilatation. The results of the measurement of 24 experimental dogs revealed that, in the experimental group, the mucosa was in good connecting condition, had a soften anastomotic stoma and a thin scar. The counts of inflammatory cells and fibroblast showed more in number at the early time after operation (Plt;0.05), while showed less in number at the advanced time of operation (Plt;0.05). In the control group, however, the mucosa were in a bad connecting condition, the scar was thicker, and the muscle layer was frequently exposed. The counts of inflammatory cells and fibroblast were fewer at the early time after operation, however, they had a clearly tendency of increasing at the anaphase after the operation. On the cytokines related to the healing of wound in the experimental group, there was a high expression and activity at the early period. There were a little expression up to postoperative 42 d. Whereas, in the control group, there had a low expression level,increased clearly on postoperative 8 d, and still a higher expression up to postoperative 42 d. Conclusions The esophagogastrostomy by layered anastomosis has a high healing quality with a thin scar. The proliferation of cells and the expression of growth factors benefits the normal healing of wound by first intention.