| Part Ⅰ Gastric Flora Analysis in Human BodyObjective: Studies have shown that many bacteria in the stomach adapt to their stomach environment and exist for a long time,and the changes of microflora in the stomach are closely related to certain digestive system diseases of the human body.In this study,two kinds of methods were adopted including traditional laboratory cultivation method and16s-r DNA detection method,in order to determine the bacterial variety in the gastric juice and whether the pathogens existed,to lay the foundation for the later study about TG-NOTES infection prevention.Methods: In this study,40 cases of gastrointestinal gastric juice were examined by routine endoscopy in the endoscopy center of Changhai Hospital from March 2015 to June 2015.Two kinds of methods were adopted to detect the bacteria.One method is to send the samples to Changhai Hospital bacteria detection section to identify the bacterial numbers and variety(20 cases).Another method is to identify the items by measuring bacteria16s-r DNA by the Shanghai Rui Yi Biological Co.,Ltd(20 cases).Results: The bacteria cultured in Changhai Hospital were mainly with Gram-positive bacteria,Gram-negative bacteria and fungi,including: Staphylococcus aureus,Neisseria gonorrhoeae,Tetranychus oryzae,Oral/Bacillus,Candida albicans and so on.16s-r DNA test results showed that the main bacterial contents of the 20 major bacteria in the gastric cavity were as follows: Prevotella,Helicobacter(Hp),Neisseria,Haemophilus,Veillonella,Fusobacterium,Porphyromonas,Streptococcus,Saccharibacteria_genera_incertae_sedis,Alloprevotella,Leptotrichia,Capnocytophaga,SR1_genera_incertae_sedis,Granulicatella,Campylobacter,Rothia,Treponema,Peptostreptococcus,Aggregatibacter,Actinomyces.Conclusion: The traditional bacterial culture method and 16s-r DNA method showed that there were bacteria in the stomach cavity,and there were many kinds of pathogens.It is clear that there is a need for local gastric cavity disinfection in the trans-gastric natural orifice transluminal endoscopic surgery(TG-NOTES).Part Ⅱ Observe the Acute Toxicity of Local Irrigation Disinfectant in RatsObjective: The preliminary study showed that gastric cavity is bacterial environment,and the localized lavage disinfection to prevent TG-NOTES infection have certified efficacy.It is reported that the local mucosal disinfectants,which are more widely used with disinfected efficacy and safety,are mainly chlorhexidine and Povidone iodine,but there is no reports related to chlorhexidine and povidone iodine in the human stomach cavity disinfection.The aim of this study is to determine whether gastric lavage chlorhexidine or povidone iodine has acute toxicity to SD rats through animal experiments,which will lay the foundation for further application in clinical research.Methods: In this research,the SD rats were treated trans-oral with chlorhexidine,povidone-iodine and normal saline(20ml/kg).Observe general situation of rats,gastric mucosa,liver and kidney tissue pathological slices and blood index(blood routine examination,liver and kidney function examination,etc.),clear the local irrigation disinfectant acute toxicity reaction of rat.Results: 1h,24 h,48h,7d,14 d,after oral administration no death was observed.There were no obvious abnormalities in the behavioral activity,appearance signs,respiration,fecal traits and local gastric mucosa.The results of hematological tests showed that there was no significant difference in blood routine indexes and biochemical indexes between the rats in the irrigation group and the control group regardless of male or female(p>0.05).Liver and kidney tissue pathological sections suggest no obvious tissue and structural damage in them.All levels of structural of gastric mucosa are integrity,no obvious bleeding and inflammatory exudation.Conclusion: Oral administration of chlorhexidine or povidone iodine has no acute toxic effect on rats.Part Ⅲ A Clinical Study of Prevent Infection with Chlorhexidine and Povidone Iodine in Trans-gastric Natural Orifice Transluminal Endoscopic Surgery(TG-NOTES)Objective: Previous studies have shown that gastric cavity is the antibacterial environment,and there is a necessity for irrigation and disinfection for it.Meanwhile,animal experiments confirmed that the localized disinfectant which chlorhexidine and povidone iodine had no acute toxicity to SD rats.The purpose of this part of the study is to clarify the changes of gastric bacteria microflora in clinical trials of human around gastric lavage with chlorhexidine and povidone iodine,clarify local gastric lavage of chlorhexidine or povidone iodine in TG-NOTES surgery whether can prevent infection,at the same time,compare the two disinfectants’ disinfection efficacy,and explicit the better one.Methods: This study used clinical trials and 40 patients were chosen which underwent routine gastroscopy from March 2015 to June 2015 in Gastroenterology of Changhai Hospital.All patients with side-lying position were applied regular intravenous anesthesia.Then the Olympus GIF-260 sterilization Endoscopy routine were used to get into the stomach cavity,to take the stomach cavity digestive juice 20ml(specimen A).The residual liquid in the stomach cavity were removed,and then chlorhexidine or povidone iodine solution 200-300 ml were used to spray irrigation,maintaining 5 min,then rinsed with100 ml of sterile saline,retaining 20 ml of rinse liquid(specimen B,specimen C).Specimens were sent for Bacteria Culture and Identification.After postoperative 24 h it was observed whether patients with abdominal pain,nausea,vomiting and other gastrointestinal symptoms.Results: All patients underwent routine gastroscopy successfully.No bacteria was detected with the method from our hospital whether it is in povidone iodine group or chlorhexidine group before irrigation and disinfection,16s-r DNA detection method results show that the type and abundance of bacterial species are significantly lower than before lavage.Significant difference: After disinfection,the type of pathogenic bacteria in the stomach cavity was significantly reduced than before,the difference between the two disinfectants group was not found in the OTU level,not the horizontal,nor found all the level difference.There was no discomfort in the chlorhexidine group and the patients in the povidone-iodine group(2/20)have nausea(2)and vomit(1),then self-mitigation without any treatment.Conclusion: using chlorhexidine solution or povidone iodine before operation is effective in preventing gastric infection by TG-NOTES.The bacteria in gastric cavity are obviously reduced after lavage,which is basically sterile.In addition,the two disinfectants are safe and effective,lower side effects.Part Ⅳ Comparative Study of Effecacy of Incision Closure between New Type Clip and Metal Clip in Trans-gastric Natural Orifice Transluminal Endoscopic Surgery(TG-NOTES)Objective: The quality of the closure of gastric wall incision is an important factor affecting the postoperative infection of TG-NOTES.In this study,we have adopted the method of animal experiment to compare the closure effective between existing simple metal clip and the new type of suture clip.Methods: 12 female experimental pigs respectively underwent conventional gastric antral site EFTR surgery,surgical diameter 3-5cm,full wall resection of gastric wall,6 cases which used metal clip clamp,6 cases which used new suture clip,comparison and evaluation the clamping time,clip quantity,survival of experimental animals.Results: After the conventional EFTR operation,the average clamping time was 8.54min;the average number of clamps was 4.16;the average clamping time of ordinary metal clip group was 10.73 min,the average number of clamps was 8.66,1 animal from ordinary metal Clip group died,other pigs are all alive.There was no significant difference in blood leukocyte and body temperature between the two groups.Conclusion: The new type of suture clip has the obvious advantages in the closed la incision,both in the clipping time and the number of clamps,and has great clin application prospect. |
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