Observation On Morphological And Histological Healing Process Of Gastrointestinal Wall After Endoscopic Full Thickness Ligation

PrefaceThe endoscopic full thickness ligation is meanly used in treating gastrointestinal(GI) submucosal tumors(diameter＜12mm) originated frommuscularis propri.The tumors were meanly including leiomyoma and interstitialoma,which were common diseases in GI system.The treatment of submucosal tumor mainly were surgical intervention,laparoscopic intervention and endoscopic ligation.Those methods respectively had their own advantages and disadvantages.Particular to the small tumors with the diameter less than 12mm,the endoscopic ligation was the first choice[3].The endoscopic ligation has the advantages as minimal invasion,short recover time and minimal financial cost.So far the endoscopic ligation for GI submucosal tumors was applied for several years,and gained a noticeable effect,which had been reported frequently.However,the ligation result was mainly followed up by esophagogastroduodenoscopy.There was not research focus on the tumor exfoliation time and healing state of the ligated lesion in a histological view.This trial was to evaluate the morphological and histological changes of the ligated mucosa,and to inspect the safety and feasibility of the endoscopic full thickness ligation.Materials1.Animal group4 experimental dogs,body weight about 15-20Kg,provided by China Medical University affiliated ShengJing hospital experimental animal department. 2.Test agentmaiyers hematoxylin;75%-100%alcohol;dimethyl benzene;1%eosin(eosin, Guangzhou Qiyun bio.Cor.);1%hydrochloric acid alcohol;PBS(citric acid buffer solution);carbazotic acid aniline red;3%hydrogen peroxide;1st antibody(Wuhan Boster cor.);2nd antibody(Sigma cor.)3.Test equipmentsendoscopic vedio system EPM-3500 and 1 gastroendoscope(Pentax EG2940), ligator:gas drived ligator(simple ring releaser),negtive aspirater,patho-clips,foreign body clips,injection syringes.4.mchemical examination equipments:microscope(OLYMPUSBX41)section cutter SHANDON finess;auto water extractor(SHANDON thermo Electron);paraffin imbedding device(LeicaEG1150H);freezer;incubator;onstant temperature bake device(TK-218VIH);paraffin section cold device(ZH-3H); electrothermal blast baking oven(HN101-2AD)Methods(1) Randomly number the 4 experimental dogs to A,B,C and D.Perform edoscopic ligation to the fundus gastricus of the dogs.Observe the healing state each day till the tumor exfoliate.Record the exfoliationg time of the ligated lesion. Respectively get the tissue sample at the exfoliation time and 3 days after exfoliation,1 week after exfoliation.Fix the tissue by 10%Formaldehyde.(2) Paraffin imbedding and section.Respectively perform HE stain,Van Gieson, immunohistological stain;SP method.(3)record and calculate the collagen content and blood capillary amount.Analyze and quantitate the records by Bandscan gray scale analysis software. Results(1) Morphological results:the average exfoliation time was 3 to 4 days.A deep ulcer was seen at the ligated region after the lesion exfoliation.It was 1 week the ulcer healing to a red scar.(2) Histological results:the collagen content and blood capillary amount enhanced obviously in 1 to 3 days after ligation,reach a peak value in 3 to 4 days,and maintain a previous value 5 days later,which meant the serosa had adhered to eachother steadly in 3-4 days after the ligation.Conclusion(1) In the endoscopic ligation treatment,before exfoliation,the serosa had adhered to eachother steadly.Thus perforation hardly appear after the ligated tissue exfoliate.(2) The local blood capillary and collagen of GI wall reach the peak value in 4 days after ligation.