| Helicobacter pylori（Hp）is an important human pathogen than can cause diffient diseases.In 2015,the Kyoto Gastritis Global Consensus blew the call for the eradication of Hp.Afterwards,the domestic and foreign guidelines for the eradication of HP continued to expand.In 2017,The fifth Chinese National Consensus Report on the management of Helicobacter pylori infection ancluded that thoes who are confirmed Hp infection should be eradicated.In 2018,Houston Consensus Conference on Testing for Helicobacter pylori Infection in the United States stated that all people infected by Hp should be treated,and in2019,Guidelines for the management of helicobacter pylori infection in japan:2016 revised edition stated that Hp infection is an indication of treatment,Antibiotics who are the main component of therapy to eradication Hp are widely used.With the overuse of antibiotics,antibiotic-related drug resistance rate is increasing,in recent years,the number of people suffering from refractory Helicobacter pylori has increased sharply.Currently,a variety of remedial treatment options have been proposed in clinical practice,and the selection of sensitive antibiotics based on the results of drug sensitivity is one of the treatment strategies.Several guidelines indicate that drug sensitivity tests are recommended if the selection of antibiotics based on clinical experience fails to eradicate Hp.However,the current drug sensitivity culture technology is still not mature enough,the sampling process requires high requirements,and the culture cycle is long,and it is difficult to be widely carried out in clinical practice,so it is necessary to explore new drug sensitivity technology.Based on this background,this study collected patients with refractory Hp infection in the Department of Gastroenterology of our hospital,and their gastric mucosa was taken and cultured by the new drug sensitivity technology,which was to explore its clinical application value.At present,the adjuvant therapy of probiotics is a hot research topic and many studies had discussed its mechanism on how to limit Hp’s growth.Some clinical studies proposed that timely addition of probiotics can restore the disturbed intestinal flora after HP infection and reduce the side effects of antibiotics.More and more datas suggest that probiotics,such as Lactobacillus and Bifidobacterium,are beneficial to Hp eradication.Probiotics’antagonistic effect on Hp may be achieved through a series of direct or indirect interactions,including secreting antimicrobial substances,competitive inhibition,interference with adhesion process,enhancement of mucosal barrier,or regulation of immunity.Microbial flora in the body’s balance is of great importance to keep the body healthy,gut microbiota is very complex,Much research focuse on the role of probiotic intestinal flora,however,gastric microbiota research is very few,we always think the stomach acid environment is no microbes,probiotics stomach micro ecological impact is rarely reported.Therefore,we presupplemented exogenous Lactobacillus in patients with refractory Hp infection to explore whether it could inhibit the colonization of Helicobacter pylori in the molecular level and adjust the microecology in the stomach,sothat it can play an auxiliary role in the eradication of Helicobacter pylori.In addition,according to statistics,China as high burden countries of gastric cancer about 89%of gastric cancer associated with Hp infection,so it is necessary to introduce"Screen and treat"strategy.However,China’s urbans who reported Hp screening rate is low,the general public’s perception of the Hp control situation is not clear,and it is also a lack of reports from different professional prevention and treatment of cognitive.Therefore,we conducted a survey of the knowledge,attitudes and practices of Chinese physicians and the general public on Hp infection,showing the current situation,and trying to explain the influencing factors of screening among the general public and the counterbalance factors of eradication treatment by doctors.Part Ⅰ The value of novel antibiotic susceptibility testing in the eradication prescription for refractory Hp infectionObjective:To study the influence of novel antibiotics susceptibility testing on the success rate of culture and eradication rate of drug-resistant strains of clinical infection;and based on the results,to analyse the resistance rates of the Hp to 5 antibiotics（amoxicillin,methazole,cramycin,levofloxacin,furanazole ketones）in patients who are multiple HP eradication failures in Shanghai.Methods:In this study,patients with refractory Helicobacter pylori infection were selected in the Department of Gastroenterology,Shanghai Changhai Hospital（Naval Medical University）from March 2019 to September 2020.Before treatment（Day 0）,the patient was examined by gastroscopy,and 1 piece of gastric antrum mucosa tissue was taken for the new drug sensitivity culture,the specific methods are as follows:gastric mucosa tissues were directly put into the new broth medium（including horse serum and brinella broth）and placed in an incubator with an ambient temperature of 37℃and a carbon dioxide concentration of 10%for 3-5 days.After being liquid turbidity,2 drops of urea solution and2 drops of phenol red solution were added to the eight pipe holes,then plus 20 ul bacterium fluid to blend for observation changes of the color,if during 10 minutes,the liquid’s color were from light yellow to red or violet,proving that Helicobacter pylori was successfully growing,if no change,the results were negative.Drug sensitivity identification are as follows:10μL bacterial liquid was inoculated into the micropores of the new drug sensitivity test plate,which containing:Amoxicillin（0.5mg/L and 1mg/L）,clarithromycin（0.5mg/L and 1mg/L）,metronidazole（4mg/L and 8mg/L）,levofloxacin（0.5mg/L and 1mg/L）,furazolidone（0.5mg/L and 1mg/L）,drug sensitivity plates were put into an incubator（the culture condition was set at 37℃,and the carbon dioxide concentration is 10%）.After 3 days,50 ul of substrate reaction solution was added to each microwell.Micropore discoloration can usually be detected within 10 minutes.If the liquid turns blue,it indicates that the Hp grows normally and it is resistant to the antibiotics.If it does not change,it indicates that the Hp is sensitive to the antibiotic.And it is moderately sensitive if the hole color changes only at low concentration.Therapies is to select two kinds of sensitive antibiotics based on the results of drug susceptibility testing combining esomeprazole and colloidal bismuth subcitrate for 10 days,then to repeat 13C or 14C breath test after stopping the medication at least one month.And finally,The success rate of culture,eradication rate and antibiotic resistance were statistically analyzed,and the correlation between different genders and ages and drug resistance rate were analyzed.Results:A total of 51 patients were included,and 33 of gastric mucosal tissue were successfully cultured,with a success rate of 64.7%.Drug resistance rates of clarithromycin,levofloxacin,metronidazole,amoxicillin,and furazolidone respectively were 81.8%,78.8%,45.5%,33.3%,and 9%.The single,double and multiple drug resistance rates in Shanghai were 12.1%,36.4%and 48.5%respectively.According to ITT analysis and PP analysis,the eradication rate was 87.9%and 93.5%.There was no statistical difference in drug resistance rates of amoxicillin,clarithromycin,levofloxacin,metronidazole and furazolidone among different genders and ages（P>0.05）.Conclusion:The success culture rate of novel antibiotics susceptibility testing was 64.7%.The single drug resistance rate and double drug resistance rate of multiple antibiotics were high in Shanghai.According to the drug sensitivity results,the eradication rate of individual treatment for refractory infection patients was high,which indicates that the reliable clinical application value of novel antibiotics susceptibility testing.Part Ⅱ The effect of compound lactobacillus acidophilus tablets on the gastric microbiota of refractory Hp infected patientsObjective:To explore whether probiotics can regulate the gastric microecological environment of patients with Hp infection at the molecular level,to assist whether probiotics can assist in the eradication of refractory Helicobacter pyloriMethods:In this study,patients who had failed to eradicate Helicobacter pylori for at least two times were selected from March 2019 to January 2020 in the Department of Gastroenterology of Shanghai Changhai Hospital.To compare the differences before and after probiotic intervention in diversity and abundance of the gastric flora of patients infecting with Hp,and to count the rate of relief of clinical symptoms in patients after probiotic intervention.Two pieces of gastric antrum mucosa were taken under endoscope before the probiotics intervention（day 0）and after the intervention（day 15）.16s DNA high-throughput sequencing was performed by extracting the DNA.Results:A total of 41 patients were included in this trial,and the results of gastric micro-ecological sequencing were as follows:（1）Alpha diversity analysis:dastric flora richness and diversity index before and after Lactobacillus intervention have no significant difference including observed species index,chaol index,PD whole tree index,goods coverage index,shannon index,simpson index（P>0.05）,showing that there is no change in the diversity and abundance of the gastric flora.（2）Beta diversity analysis:Uni Frac heat map analysis,Anosim analysis,species Adnois analysis,species MRPP analysis,NMDS analysis and PCOA analysis showed similarities between lactobacillus intervention and gastric flora before and after the intervention,they are no significant separation,indicating there is no significant differences in species diversity before and after Lactobacillus intervention.（3）LEFSE analysis:It was found that the relative abundance of Dorea,Faecalibacterium,Corynebacterium and Corynebacterium increased significantly after probiotic intervention.（4）Clinical symptoms:Clinical symptoms improved significantly after probiotic intervention（P<0.001）.Conclusion:Lactobacillus pre-intervention for 2 weeks has little effect on the relative abundance and species diversity of gastric flora,which may mainly play a role by regulating certain gastric flora and intestinal flora.Part Ⅲ The survey of public awareness of Helicobacter pylori infection in ChinaObjectives:The purpose of this study was to conduct a national survey on the knowledge,attitudes,and practices of physicians and the general public regarding Helicobacter pylori infection in China,and to present the current situation。Methods:This was an Internet-based survey of the general Chinese population and Chinese physicians from different specialties,carried out from January to February 2019.Both surveys（general population’s and physicians’）included questions to assess the knowledge and attitudes toward H pylori and its action.Data analysis was performed using SPSS 20.0.Results:A total of 3211 people and 546 physicians were enrolled.In the population,the proportion of subjects who answered correctly to all questions about H pylori’s infectivity was only 16%,and that for H pylori’s harmfulness and that for H pylori preventive measures were 35%and 43.6%,respectively.In general,physicians had a better understanding of H pylori’s harmfulness（83.9%）than the other population.The vast majority of the surveyed population（87.0%）and physicians（82.2%）supported a national H pylori screening plan to prevent gastric cancer.The support ratio paralleled with the overall knowledge level.Unexpectedly,gastroenterologists tend to have a relatively low support rate for H pylori screening than non-gastroenterologists（58.2%vs 84.2%,P<.001）,which may be related to consideration of heavy medical burden（67.3%）.Conclusion:The awareness rate and screening rate of HP in Chinese people are low,especially in provinces with high incidence and mortality of gastric cancer.Health education of HP knowledge should be strengthened.The general public holds a positive attitude towards HP screening,which is conducive to the development of HP screening and radical cure,and can further reduce the burden of gastric cancer in China.Chinese physicians of all specialties have a high level of HP knowledge and generally support universal HP screening,but gastroenterologists have a low support for HP screening,which may be related to the lack of medical resources,suggesting that health departments should strengthen resource investment related to HP screening. |
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