Increased VGLUT3 involved in visceral hyperalgesia in a rat model of irritable bowel syndrome

AIM:To investigate the activity of vesicular glutamate transporter-3(VGLUT3) in a visceral hyperalgesia rat model of irritable bowel syndrome,and the role of mast cells(MCs).METHODS:Transient intestinal infection was inducedby oral administration of Trichinella spiralis larvae in rats.On the 100th day post-infection(PI),the rats were divided into an acute cold restraint stress(ACRS)group and a non-stressed group.Age-matched untreated rats served as controls.The abdominal withdrawal reflex was used to measure the visceromotor response to colorectal distension(CRD).The expression levels of VGLUT3 in peripheral and central neurons were analyzed by immunofluorescence and western blotting.RESULTS:VGLUT3 expression in the L6S1 dorsal root ganglion cells was significantly higher in the PI group than in the control group(0.32±0.009 vs0.22±0.008,P<0.01),and there was no significant difference in the expression of VGLUT3 between MCdeficient rats and their normal wild-type littermates.Immunofluorescence showed that the expression levels of VGLUT3 in PI+ACRS rats were enhanced in the prefrontal cortex of the brain compared with the control group.CONCLUSION:VGLUT3 is involved in the pathogenesis of visceral hyperalgesia.Coexpression of c-fos,5-hydroxytryptamine and VGLUT3 after CRD was observed in associated neuronal pathways.Increased VGLUT3 induced by transient intestinal infection was found in peripheral nerves,and was independent of MCs.Moreover,the expression of VGLUT3 was enhanced in the prefrontal cortex in rats with induced infection and stress.

Gene expression profiles in peripheral blood mononuclear cells of ulcerative colitis patients

AIM: To identify peripheral blood mononuclear cell (PBMC) gene expression profiles of ulcerative colitis (UC) patients, using oligonucleotide microarrays, to gain insights into UC molecular mechanisms. METHODS: The Human OneArray microarrays were used for a complete genome-wide transcript profiling of PBMCs from 12 UC patients and 6 controls. Differential analysis per gene was performed with a random variance model; t test and P values were adjusted to control the false discovery rate (5%). Gene ontology (GO) was deployed to analyze differentially expressed genes at significant levels between patients and controls to identify the biological processes involved in UC. RESULTS: Comparative analysis revealed that 4438 probes (4188 genes) were differentially expressed between the two groups, of which 3689 probes (3590 genes) were down-regulated whereas 749 probes (598 genes) were up-regulated. Many disregulated genes in our data have been reported by previous microarray studies carried out on intestinal mucosa samples, such as S100A8 , CEACAM1 and S100A9 . GO enrichment analysis revealed 67 high enrichment up-regulated categories and one significant down-regulated category. The up-regulated genes were mainly involved in immune and inflammatory response, cell cycle and proliferation, DNA metabolism and repair. CONCLUSION: Gene expression profiling of PBMCs from patients with UC has highlighted several novel gene categories that could contribute to the pathogenesis of UC.

Prognostic prediction models for postoperative patients with stageⅠtoⅢcolorectal cancer based on machine learning

BACKGROUND Colorectal cancer(CRC)is characterized by high heterogeneity,aggressiveness,and high morbidity and mortality rates.With machine learning(ML)algorithms,patient,tumor,and treatment features can be used to develop and validate models for predicting survival.In addition,important variables can be screened and different applications can be provided that could serve as vital references when making clinical decisions and potentially improving patient outcomes in clinical settings.AIM To construct prognostic prediction models and screen important variables for patients with stageⅠtoⅢCRC.METHODS More than 1000 postoperative CRC patients were grouped according to survival time(with cutoff values of 3 years and 5 years)and assigned to training and testing cohorts(7:3).For each 3-category survival time,predictions were made by 4 ML algorithms(all-variable and important variable-only datasets),each of which was validated via 5-fold cross-validation and bootstrap validation.Important variables were screened with multivariable regression methods.Model performance was evaluated and compared before and after variable screening with the area under the curve(AUC).SHapley Additive exPlanations(SHAP)further demonstrated the impact of important variables on model decision-making.Nomograms were constructed for practical model application.RESULTS Our ML models performed well;the model performance before and after important parameter identification was consistent,and variable screening was effective.The highest pre-and postscreening model AUCs 95%confidence intervals in the testing set were 0.87(0.81-0.92)and 0.89(0.84-0.93)for overall survival,0.75(0.69-0.82)and 0.73(0.64-0.81)for disease-free survival,0.95(0.88-1.00)and 0.88(0.75-0.97)for recurrence-free survival,and 0.76(0.47-0.95)and 0.80(0.53-0.94)for distant metastasis-free survival.Repeated cross-validation and bootstrap validation were performed in both the training and testing datasets.The SHAP values of the important variables were consistent with the clinicopathological characteristics of patients with tumors.The nomograms were created.CONCLUSION We constructed a comprehensive,high-accuracy,important variable-based ML architecture for predicting the 3-category survival times.This architecture could serve as a vital reference for managing CRC patients.

Characteristic dysbiosis of gut microbiota of Chinese patients with diarrhea-predominant irritable bowel syndrome by an insight into the pan-microbiome

Background:Irritable bowel syndrome(IBS)is reported associated with the alteration of gut microbial composition termed as dysbiosis.However,the pathogenic mechanism of IBS remains unclear,while the studies of Chinese individuals are scarce.This study aimed to understand the concept of dysbiosis among patients with Chinese diarrhea-predominant IBS(IBS-D),as a degree of variance between the gut microbiomes of IBS-D population and that of a healthy population.Methods:The patients with IBS-D were recruited(assessed according to the Rome III criteria,by IBS symptom severity score)from the Outpatient Department of Gastroenterology of Peking University Third Hosp让al,and volunteers as healthy controls(HCs)were enrolled,during 2013.The 16S rRNA sequences were extracted from fecal samples.Ribosomal database project resources,basic local alignment search tool,and SparCC software were used to obtain the phylotype composition of samples and the internal interactions of the microbial community.Herein,the non-parametric test,Wilcoxon rank-sum test was carried out to find the statistical significance between HC and IBS-D groups.All the P values were adjusted to q values to decrease the error rate.Results:The study characterized the gut microbiomes of Chinese patients with IBS-D,and demonstrated that the dysbiosis could be characterized as directed alteration of the microbiome composition leading to greater disparity between relative abundance of two phyla,Bacteroidetes(Z=4.77,q=1.59×10^-5)and Firmicutes(Z=-3.87,q=5.83×10^-4).Moreover,it indicated that the IBS symptom features were associated with the dysbiosis of whole gut microbiome,instead of one or several certain genera even they were dominating.Two genera,Bacteroides and Lachnospiracea incertae sedis,were identified as the core genera,meanwhile,the non-core genera contribute to a larger pan-microbiome of the gut microbiome.Furthermore,the dysbiosis in patients with IBS-D was associated with a reduction of network complexity of the interacted microbial community(HC us.IBS-D:639 vs.154).The disordered metabolic functions of patients with IBS-D were identified as the potential influence of gut microbiome on the host(significant difference with q<0.01 between HC and IBS-D).Conclusions:This study supported the view of the potential influence of gut microbiome on the symptom of Chinese patients with IBS-D,and further characterized dysbiosis in Chinese patients with IBS-D,thus provided more pathological evidences for IBS-D with the further understanding of dysbiosis.

A population-based survey of gastroesophageal reflux disease in a region with high prevalence of esophageal cancer in China

Background: The exact relationship between gastroesophageal reflux disease (GERD) and esophageal squamous cell cancer (ESCC) is far from clarification. The aim of this study was to investigate the epidemiology of GERD in a region with high prevalence of ESCC in China. Methods: A population-based, cross-sectional study was conducted in a high ESCC prevalent area, Anyang, Henan, China. All subjects fulfilled questionnaires and underwent gastroendoscopy with routine esophageal biopsy. The subjects were divided into GERD subtypes (reflux esophagitis [RE] and non-erosive reflux disease [NERD]) and controls. Odds ratios (ORs) and 95% confidence intervals (Cis) were calculated to examine risk factors for RE and NERD. Results: A total of 2844 subjects were finally enrolled. The prevalence of GERD (RE + NERD) was 17.3%. Among them, 271 (9.53%) adults were diagnosed with RE. The prevalence of RE increased with age (7.09% in 45-50 years, 8.00% in 51-60 years, and 9.53% in 61-69 years,x^2 = 62.216, P < 0.001). Sixty-seven (2.36%) subjects were diagnosed with the silent RE. A total of 221 (7.77%) subjects were diagnosed with NERD. Frequent liquid food consumption (OR [95% CI]: 1.502 [1.076-2.095]) was independent risk factor for RE as well as age, male gender, high body mass index (BMI), ever smoking. Age was independent risk factor for NERD. For silent RE, age, male gender, and frequent liquid food consumption were risk factors. Conclusions: In the population with high prevalence of ESCC, a high prevalence of GERD and inverted proportion of RE/NERD were presented. Age was an independent risk factor for GERD. The male gender, high BMI, smoking, and frequent liquid food consumption may be risk factors for RE but not for NERD.

Epidemiological and clinical features of functional dyspepsia in a region with a high incidence of esophageal cancer in China

Background:Functional dyspepsia(FD)has rarely been investigated in areas with a high prevalence of esophageal squamous cell carcinoma(ESCC).This study aims to reveal the epidemiological and clinical features of FD and organic dyspepsia(OD)in such a population.Methods:A middle-aged and elderly population-based study was conducted in a region with a high incidence of ESCC.All participants completed the Gastroesophageal Reflux Disease Questionnaire and Functional Gastrointestinal Disease Rome III Diagnostic Questionnaire,and they underwent gastroscopy.After exclusion of gastroesophageal reflux disease,uninvestigated dyspepsia(UID)was divided into OD and FD for further analyses.Results:A total of 2916 participants were enrolled from July 2013 to March 2014 in China.We detected 166 UID cases with questionnaires,in which 17 patients with OD and 149 with FD were diagnosed via gastroscopy.OD cases presented as reflux esophagitis(RE),ESCC,and duodenal ulcer.Heartburn(52.94%)and reflux(29.41%)were common in OD,but no symptomatic differences were found between FD and OD.Male sex,low education level,and liquid food were the risk factors for OD,while frequent fresh vegetable consumption was a protective factor.FD included 56(37.58%)cases of postprandial distress syndrome(PDS),52(34.89%)of epigastric pain syndrome(EPS),nine(6.04%)of PDS+EPS,and 32(21.48%)of FD+functional esophageal disorders.The Helicobacter pylori infection rate in FD patients was not higher than that in the control group(34.23%vs.42.26%,P=0.240).Frequent spicy food consumption was associated with PDS(odds ratio[OR]:2.088,95%confidence interval[CI]:1.028–4.243),while consumption of deep well water was protective for PDS(OR:0.431,95%CI:0.251–0.741).Conclusions:The prevalence of FD was 5.11%in the studied population.Gastroscopy should be prescribed for dyspepsia patients in case that ESCC and RE would be missed in UID cases diagnosed solely by the Rome III questionnaire.Trial Registration:ClinicalTrials.gov,NCT01688908;https://clinicaltrials.gov/ct2/show/record/NCT01688908.

Effect of the interleukin 10 polymorphisms on interleukin 10 production and visceral hypersensitivity in Chinese patients with diarrhea-predominant irritable bowel syndrome

Background:Irritable bowel syndrome (IBS),a functional gastrointestinal disorder,is characterized by cytokine imbalance.Previously,decreased plasma interleukin 10 (IL-10) level was reported in patients with IBS,which may be due to genetic polymorphisms.However,there are no reports correlating the IL-10 polymorphisms with IL-10 production in patients with IBS.This study aimed to analyze the effect of IL-10 polymorphisms on IL-10 production and its correlation with the clinical symptoms in Chinese patients with diarrhea-predominant IBS (IBS-D).Methods:Two IL-10 single nucleotide polymorphisms (rs1800871 and rs1800896) were detected in 120 patients with IBS-D and 144 healthy controls (HC) using SNaPshot.IBS symptom severity score,Bristol scale,hospital anxiety,and depressive scale (HADS) were used to evaluate the clinical symptoms,as well as the psychological status and visceral sensitivity of the subjects.IL-10 levels in the plasma and peripheral blood mononuclear cell (PBMC) culture supernatant were measured using enzyme-linked immunosorbent assay,while those in ileal and colonic mucosal biopsies were measured using immunohistochemistry.Results:The frequency of rs1800896 C allele was significantly lower in the patients with IBS-D than that in the HC (odds ratio:0.49,95% confidence interval:0.27-0.92,P =0.0240).The IL-10 levels in the plasma (P =0.0030) and PBMC culture supernatant (P =0.0500) of the CT genotype subjects were significantly higher than those in the TT genotype subjects.The CT genotype subjects exhibited a higher pain threshold in the rectal distention test than the TT genotype subjects.Moreover,IL-10 rs1800871 GG genotype subjects showed an increase in the HADS score compared to other genotype subjects.Conclusions:IL-10 rs1800896 C allele is correlated with higher IL-10 levels in the plasma and the PBMC culture supernatant,which is associated with a higher pain threshold in the Chinese patients with IBS-D.This study provides an explicit relationship of IL-10 polymorphisms with IL-10 production,which might help in understanding the pathogenesis of IBS-D.

Research gaps for three main tropical diseases in the People’s Republic of China

This scoping review analyzes the research gaps of three diseases:schistosomiasis japonica,malaria and echinococcosis.Based on available data in the P.R.China,we highlight the gaps between control capacity and prevalence levels,and between diagnostic/drug development and population need for treatment at different stages of the national control programme.After reviewing the literature from 848 original studies and consultations with experts in the field,the gaps were identified as follows.Firstly,the malaria research gaps include(i)deficiency of active testing in the public community and no appropriate technique to evaluate elimination,(ii)lack of sensitive diagnostic tools for asymptomatic patients,(iii)lack of safe drugs for mass administration.Secondly,gaps in research of schistosomiasis include(i)incongruent policy in the implementation of integrated control strategy for schistosomiasis,(ii)lack of effective tools for Oncomelania sp.snail control,(iii)lack of a more sensitive and cheaper diagnostic test for large population samples,(iv)lack of new drugs in addition to praziquantel.Thirdly,gaps in research of echinococcosis include(i)low capacity in field epidemiology studies,(ii)lack of sanitation improvement studies in epidemic areas,(iii)lack of a sensitivity test for early diagnosis,(iv)lack of more effective drugs for short-term treatment.We believe these three diseases can eventually be eliminated in China's Mainland if all the research gaps are abridged in a short period of time.

Clinical characteristic and fecal microbiota responses to probiotic or antidepressant in patients with diarrhea-predominant irritable bowel syndrome with depression comorbidity: a pilot study

To the Editor: Irritable bowel syndrome (IBS) is a prevalent functional gastrointestinal disorder that presents as abdominal pain with altered bowel habits. The pathophysiologic mechanism of IBS is not well understood, although many hypotheses have been proposed, including visceral hypersensitivity, gastrointestinal dysmotility, lowgrade inflammation of the intestinal mucosa, and dysfunction of the brain-gut interaction. Dysfunction of the braingut interaction is thought to be involved in IBS because a considerable proportion of patients with IBS have some form of psychologic comorbidity, such as depression or anxiety. In addition, mental and psychologic problems increase the risk for IBS and the symptom severity of IBS. Recently, changes in the gut microbiota have also been suggested to contribute to both IBS and depression. Our previous study demonstrated significant altered gut microbiota profiles in patients with diarrhea-predominant IBS (IBS-D) and depression.

Synthesis of 2,5-disubstitued benzimidazole using SnCl_2-catalyzed reduction system at room temperature

Stannous chloride dihydrate is used as an efficient catalyst in reductive cyclization of 2-nitro-5- substituted aniline Schiff base leading to stable 2,5-disubstitued benzimidazole derivatives in excellent yields with good purity. It provides a novel method of synthesis of 2,5-disubstitued benzimidazole under reductive system at room temperature.