| Objective:To study the possibility of seromuscular anastomosis of colon in rats without sewing submucosa and the healing quality of colon compared with the traditional manual method---single-layer anastomosis.Methods:48 healthy and clean Wistar rats, female, weight 240-270g. Randomly divided into two groups:single-layer anastomosis group(group A, n=24) and seromuscular anastomosis group(group B, n=24), single-cage rearing. Right colon was cut and end-to-end anastomosis was performed on each rat. 8 rats of each group were randomly chosen and killed on postoperative days 3,7 and 14. Harvest the tissues around the colon anastomosis.1.Determination of the peritoneal adhesion index.2.Determination of the anastomotic bursting pressure (ABP).3.Determination of the colonic anastomotic hydroxyproline (HP) content.4.HE staining, histopathologic evaluation of the anastomotic sites.5.CD146 immunohistochemical staining of the anastomotic sites.6.Determination of MVD counting.7.Immunohistochemical staining of the anastomotic sites using type I collagen.8.Determination of the relative expression of CD146mRNA using RT-PCR.9.Determination of the relative expression of CD146 protein using Western blot.10. Measurement data was presented as mean±standard deviation(x± s) and the single factor analysis of variance P<0.05 was considered statistically significant.Results:1.Peritoneal adhesion index:The determination showed on day 3,7 and 14, the peritoneal adhesion index of group B was apparently lower than that of group A, with statistical significance(P<0.05).2.Anastomotic bursting pressure(ABP):On day 3, ABP of group B was slightly lower than that of group A, but without statistical significance(P> 0.05). On day 7, ABP of group B was higher, but without statistical significance(P> 0.05). On day 14, ABP of the two groups was close(.P> 0.05).3.Hydroxyproline(HP) content:The hydroxyproline content of postoperative colonic anastomotic tissue upward trended over time. On day 3. HP content of the two groups was close (P> 0.05). On day 7 and 14 HP of group B was apparently higher than that of group A(P<0.05).4. HE staining:Histopathologic evaluation of the anastomotic sites indicated that on day 3 neutrophil of group A was more than that of group B, with a large number of necrotic cells arranged disorderly. On day 7, neutrophil declined while mononuclear cells and lymphocytes increased. Granulation tissue of group B formed, in which a large number of fibroblasts and new blood vessels vertically grew against wound, more than those of group A. On day 14, the healing of group B was better than that of group A, with cells arranged orderly.5. CD146 immunohistochemical staining:Observations of pathological slices indicated that on day 3, CD146 expression of endothelial cells of group B was positive, and strongly positive on day 7, while it reduced the positive degree on day 14; CD146 expression of endothelial cells of group A was some positive, some weakly positive on day 3, and positive on day 7 and 14.6. MVD value:On day 3, the MVD value of group B significantly increased, reached a high level on day 7, then decreased on day 14 to a lower level. On day 3 and 14, the MVD value of group A was slightly lower than that of group B, but without statistical significance(P> 0.05); while on day 7, it was apparently lower(P<0.05). 7. The area value of type I collagen:On day 3 there was no significant difference between the two groups(P> 0.05). And on day 7 and 14, the area value of type I collagen of group B significantly increased, and was significantly different(P<0.05).8. The relative expression of CD146mRNA:On day 3, there was no statistical difference on the relative expression of the two groups(P> 0.05). And on day 7 and 14, the relative expression of group B significantly increased, with larger difference from that of group A(P<0.05).9. The relative expression of CD146 protein:On day 3, there was no statistical difference on the relative expression of the two groups(P> 0.05). On day 7, the relative expression of CD 146 protein of group B significantly increased, with larger difference from that of group A(P<0.05). While on day 14, the relative expression of CD146 in both groups significantly reduced, without any significant difference(P> 0.05).Conclusion:l.The peritoneal adhesion caused by seromuscular anastomosis of colon is slighter than that caused by single-layer anastomosis.2.The seromuscular anastomosis of colon is as safe and reliable as the single-layer anastomosis.3.The hydroxyproline content of seromuscular anastomosis group increases compared with that of single-layer anastomosis.4. The content of type I collagen of seromuscular anastomosis group increases compared with that of single-layer anastomosis.5. The number of blood vessels of seromuscular anastomosis group increases compared with that of single-layer anastomosis.6. Seromuscular anastomosis of colon is safe and reliable and easier than any other manual anastomosis.
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