Qualitative and quantitative analyses of the bifidobacterial microbiota in the colonic mucosa of patients with colorectal cancer, diverticulitis and inflammatory bowel disease

AIM： To characterize the bifidobacterial microbiota of the colonic mucosa in patients with colon cancer, inflammatory bowel disease or diverticulitis. METHODS： A sample of the distal colonic mucosa was taken during surgery from a total of 34 patients, twenty-one with diagnosed colorectal cancer, nine with diverticulitis and four with inflammatory bowel disease, requiring surgery for their condition. Bacterial DNA was extracted from the resected mucosal samples and bifidobacterial mucosa-associated microbiota was qualitatively and quantitatively determined by means of qualitative and quantitative PCR. RESULTS： Bifidobacteria were found in 100% of the samples from patients with diverticulitis or IBD and a 76% of those suffering colon cancer. The species B. Iongum and B. bifidum were the most widely found, followed by B. animalis, B. catenulatum and B. adolescentis. B. breve, B. dentium and B. angulatum were not detected in any sample. A significantly higher occurrence of B. Iongum was observed in patients with diverticulitis than in those with colon cancer or IBD （100%, 62% and 75%, respectively, P 〈 0.05）. Similar results were obtained for B, animalis （56%, 0% and 25%, P 〈 0.05）, while B. adolescentis was only found in the mucosa from patients with colon cancer （5 out of 21, 24%）. At the quantitative level, patients with colon cancer or IBD showed lower counts of total Bifidobacterium （4.94 and 5.91 vs 6.96 log Cells/sample, respectively, P 〈 0.05） and of the species B. longum （4.05 and 4.79 vs 6.76, P 〈 0.05） than those with diverticulitis.CONCLUSION： Aberrancies in mucosa associated microbiota are present in different intestinal diseases. This may indicate a role of the microbiota in the pathogenesis of these diseases.

Effects of moxibustion on heat-shock protein 70 expression in the spinal cord and colonic mucosa in a rat model of ulcerative colitis

Pathological changes in the colon are closely associated with the spinal cord, and innervation of spinal cord can regulate cellular functions. Our previous studies verified that moxibustion protects and restores the colonic mucosa, but the mechanisms of action remain unknown. The present study observed the effects of moxibustion and salicylazosulfapyridine on expression of heat-shock protein 70 （HSP70） and its mRNA in the spinal cord and colonic mucosa of ulcerative colitis rats. Results demonstrated that moxibustion and salicylazosulfapyridine increased HSP70 mRNA expression in the spinal cord and colonic mucosa of ulcerative colitis rats. The decreased transcriptional activity of HSP70 in the spinal cord and colonic mucosa might participate in damage to the colonic mucosa in ulcerative colitis rats. Moxibustion exerted protective effects on colonic mucosa by up-regulating HSP70 transcriptional activity in the spinal cord and colonic mucosa.

Proteomic analysis of down-regulated proteins in colonic mucosa of chronic slow transit constipation rats

Objective： To investigate the alternations of proteins in the colonic mucosa of chronic slow transit constipation （STC） rats with a 2-DE-based proteomic method and analyze the function of these down-regulated proteins so as to provide theoretical basis for the pathogenesis of intestinal mucosa of chronic STC rats. Methods： STC model was established by feeding rats with 8 mg/（kg＇d） diphenoxylate for 120 d. An experimental model of chronic STC rat was used for separation of proteomics from colonic mucosa using two-dimensional electrophoresis （2-DE）. Proteins altered in expressional level were identified by Image Master 2DElite, mass spectrometry, and bibliometrics were applied to identify the differential protein expression and their clinical s observed in the pathogenesis lgn of ificance and function were analyzed. Results： Obvious differential protein expression was STC, including mast cell protease （A1）, non-specific dipeptidase （A2） and chondrosome succinate dehydrogenase precursor （A3）. The expressions of A1, A2 and A3 were down-regulated in the gel graph of STC rats Conclusion： The down-regulation of chondrosome succinate dehydrogenase, mast cell protease as well as non-specific dipeptidase in rat colon suggests the functional impairment of the oxidoreduction of mitochondrion is very important in the genesis and development of STC. The immunological reaction of STC rats is weakened, and the function of digesting and absorbing protein may be damaged to some extent.

DISTINGUISHING BETWEEN HYPERPLASTIC AND ADENOMATOUS POLYPS AND NORMAL COLONIC MUCOSA BY USING MULTIPHOTON LASER SCANNING MICROSCOPY

Precisely distinguishing between hyperplastic and adenomatous polyps and normal human colonic mucosa at the cellular level is of great medical significance.In this work,multiphoton laserscarming microscopy(MPLSM)was used to obtain the high.-contrast images and the morpho-logical characteristics from normal colonic mucosa,hyperplastic polyps and tubular adenoma.Byintegrating the length and area measurement tools and computing tool,we quantified thedifference of crypt morphology and the alteration of nuclei in normal and diseased human colonicmucosa.Our results demonstrated that the morphology of crypts had an obvious tendency tocystic dilatation or elongated in hyperplastic polyps and tubular adenoma.The cont ent andnumber of mucin droplets of the scattered goblet cells had a piecemeal reduction in hyperplastic polyps and a large decrease in tubular adenoma The nuclei of epithelial cells might be elongated and pseudostratified,but overt dysplasia was absent in hyperplastic polyps.Nevertheless,thenuclei showed enlarged,crowded,stratified and a rod-like structure,with loss of polarity intubular adenoma.These results suggest that MPLSM has the capacity to distinguish betweenhyperplastic and adenomat ous polyps and normal human colonic mucosa at the celular level.

MicroRNA exhibit altered expression in the inflamed colonic mucosa of ulcerative colitis patients

To investigate the miRNA expression in colonic mucosal biopsies from endoscopically inflamed and non inflamed regions of ulcerative colitis (UC) patients. METHODSColonic mucosal pinch biopsies were analyzed from the inflamed and non inflamed regions of same UC patient. Total RNA was isolated and differential miRNA profiling was done using microarray platform. Quantitative Real Time PCR was performed in colonic biopsies from inflamed (n = 8) and non-inflamed (n = 8) regions of UC and controls (n = 8) to validate the differential expression of miRNA. Potential targets of dysregulated miRNA were identified by using in silico prediction tools and probable role of these miRNA in inflammatory pathways were predicted. RESULTSThe miRNA profile of inflamed colonic mucosa differs significantly from the non-inflamed. Real time PCR analysis showed that some of the miRNA were differentially expressed in the inflamed mucosa as compared to non inflamed mucosa and controls (miR-125b, miR-223, miR-138, and miR-155), while (miR-200a) did not show any significant changes. In contrast to microarray, where miR-378d showed downregulation in the inflamed mucosa, qRT-PCR showed a significant upregulation in the inflamed mucosa as compared to the non inflamed. The in silico prediction analysis revealed that the genes targeted by these miRNAs play role in the major signaling pathways like MAPK pathway, NF-κB signaling pathway, cell adhesion molecules which are all assciated with UC. CONCLUSIONThe present study reports disease specific alteration in the expression of miR-125b, miR-155, miR-223 and miR-138 in UC patients and also predict their biological significance.

Histopathological changes of free buccal mucosa and colonic mucosa grafts after translation to dog bladder

Over the past years, more cases using buccal mucosa for urethral reconstruction have been reported.(1-4) The excellent early results with this tissue led some authors to extend their indications for its use. However, patients with complex, long-segment urethral strictures and significant scar tissue formation after the failure of previous urethroplasty, still present an operative challenge. The buccal mucosa may not be useful for the treatment of the complicated lengthy urethral strictures because of limited material. To explore the possibility of urethral reconstruction with a graft of colonic mucosa for the treatment of complicated lengthy strictures, we investigated the use of colonic mucosa as a novel substitute for urethral reconstruction in dogs with severe lengthy urethral structure.(5,6) The objects of the present study were to investigate further whether the colonic mucosal graft is an ideal substitute material, for urethral reconstruction, in place of buccal mucosal graft and their pathological characteristic after long-term exposure to urine.

Colon mucosal injury caused by water jet malfunction during a screening colonoscopy:A case report

BACKGROUND Screening colonoscopies are routinely performed and have low occurrences of adverse events such as perforation,bleeding,infection,and post-polypectomy syndrome.True device related adverse events are rarely reported in the literature.CASE SUMMARY We report a case of a 51-year-old patient without past medical history who presented for her first screening colonoscopy.The patient was thought to have friable mucosa in the cecum and oozed upon water irrigation during screening colonoscopy.It was later identified that the colonoscope used during the index procedure had malfunctioned and produced a pin-point water jet which damaged the colon mucosa of cecum.The maintenance service identified a piece of rubber fragment lodged in the instrument component at the tip of the scope,resulting in high pressure water jet.Repeat colonoscopy with a functioning colonoscope confirmed normal colon mucosa without friability.CONCLUSION This is the first report of mucosa injury from a colonoscope water jet malfunction.Endoscopists should recognize the potential for endoscopic malfunction.

Anti-inflammatory mechanism of oxymatrine in dextran sulfate sodium-induced colitis of rats

AIM： To investigate the anti-inflammatory mechanism of oxymatrine in dextran sulfate sodium （DSS）-induced colitis of rats. METHODS： Acute colitis was induced by giving 2% DSS orally in drinking water for 8 d. Twenty-six male rats were randomized into oxymatrine-treated group （group A, 10 rats）, DSS control （group B, 10 rats） and normal control （group C, 6 rats）. The rats in group A were injected muscularly with oxymatrine at the dosage of 63 mg/（kg·d） from d 1 to 11 and drank 2% DSS solution from d 4 to 11. The rats in group B were treated with 0.9% saline in an equal volume as group A and drank 2% DSS solution from d 4 to 11. The rats in group C were treated with 0.9% saline as group B from d 1 to 11 and drank water normally. Diarrhea and bloody stool as well as colonic histology were observed. The levels of serum tumor necrosis factor-α （TNF-α） and interleukin-6 （IL-6） were determined by ELISA, and nuclear factor-κB （NF-κB） activity and the expression of inter-cellular adhesion molecule-1 （ICAM-1） in colonic mucosa were detected by immunohistochemistry method. RESULTS： Compared with DSS control group, the inflammatory symptoms and histological damages of colonic mucosa in oxymatrine-treated group were significantly improved, the serum levels of TNF-α, IL-6, and the expression of NF-κB, ICAM-1 in colonic mucosa were significantly reduced. CONCLUSION： The fact that oxymatrine can reduce the serum levels of TNF-α, IL-6, and the expression of NF-κB and ICAM-1 in colonic mucosa in DSS-induced colitis of rats indicates that oxymatrine may ameliorate the colonic inflammation and thus alleviate diarrhea and bloody stool.