Effects of Estrogen on Mucosal Structure and Numbers and Distribution of Intraepithelial Lymphocytes and Goblet Cells in Small Intestine of Rats

To study the effects of estrogen on the structure of the intestinal mucosal barrier, 18 healthy female Wistar Rats underwent estrus synchronization. In diestrus, they were divided into three groups： one sham operated control group （SHAM） ; one ovariec- tomized group （OVX） ; and one ovariectomized plus estradiol benzoate group （ OVX ＋ E2 ）. Intestinal mu- cosal epithelial cells, intraepithelial lymphocytes （[EL）, and goblet cells （GCs） were observed by light microscope. The results showed that in the OVX group, the intestinal mucosa damaged obviously, the villus atrophied, the ratio of villus height to crypt depth reduced, and the number of IELs and GCs re- duced. The indicators of OVX ＋ Ez group were signif- icantly higher than OVX group, but some indicators were lower than SHAM. These indicated that the function of intestinal mucosal barrier was greatly dam- aged in ovariectomied rat, and proper dosage of estra- diol benzoate Would improve the function of small in- testinal mucosal barrier in ovariectomied rat to some degree.

Effects of Chinese Medicine on the Number of Intraepithelial Lymphocytes and Goblet Cells of Intestinal Villus of Heat Stress Layers

[ Objective] To study the effects of Chinese herbal additives on the number of intraepithelial lymphocyte and goblet cells of intestinal villus of heat stress layers. [Method] 180 healthy 88-day-old ISA brown egg roosters were selected and randomly divided into nine treatment groups, nor- mal temperature control group, high temperature control group, VC group, prescription one high-dose group, prescription one middle-dose group, prescription one low-dose group, prescription two high-dose group, prescription two middle-dose group, prescription two low-dose group, respec- tively. Prescription one and two groups were respectively fed with low, medium and high concentrations of the three doses of Chinese herbal ex- tracts, and VC group was fed with the VC in aqueous solution. Histological sections conventional technology and HE staining method were used to observe the number of intraepithelial lymphocytes and goblet cells in each sections of small intestine of chicken. [ Result] The number of chicken in- testinal epithelial lymphocytes and goblet cells showed a gradually decreasing trend in a high-temperature state. [ Conclusion] Prescription one and two groups could promote the cytopoiesis of goblet cells and lymphocytes, and the effect of prescription two was the best. Moreover, Adding the Chinese herbs had good effects on relieving the heat stress of layers.

Time-dependent impact of a high-fat diet on the intestinal barrier of male mice

BACKGROUND Excessive saturated fat intake compromises the integrity of the intestinal mucosa,leading to low-grade inflammation,impaired mucosal integrity,and increased intestinal permeability,resulting in the migration of lipopolysaccharide(LPS)to other tissues.AIM To evaluate the chronic effects(at 10 and 16 wk)of a high-fat diet(HFD)(with 50%energy as fat)on the phylogenetic gut microbiota distribution and intestinal barrier structure and protection in C57BL/6 mice.METHODS Forty adult male mice were divided into four nutritional groups,where the letters refer to the type of diet(control and HFD or HF)and the numbers refer to the period(in weeks)of diet administration:Control diet for 10 wk,HFD for 10 wk,control diet for 16 wk,and HFD for 16 wk.After sacrifice,biochemical,molecular,and stereological analyses were performed.RESULTS The HF groups were overweight,had gut dysbiosis,had a progressive decrease in occludin immunostaining,and had increased LPS concentrations.Dietary progression reduced the number of goblet cells per large intestine area and Mucin2 expression in the HF16 group,consistent with a completely disarranged intestinal ultrastructure after 16 wk of HFD intake.CONCLUSION Chronic HFD intake causes overweight,gut dysbiosis,and morphological and functional alterations of the intestinal barrier after 10 or 16 wk.Time-dependent reductions in goblet cell numerical density and mucus production have emerged as targets for countering obesity-driven intestinal damage.

Goblet cell carcinoid of the appendix and mixed adenoneuroendocrine carcinoma: Report of three cases

Neuroendocrine neoplasms are the most common epithelial tumors among appendix tumors. Appendix tumors that are completely or partially composed of neuroendocrine cells are divided into two categories: Classic carcinoid tumors and goblet cell carcinoid tumors(GCCT). They are known to progress more aggressively than classic(neuro) endocrine tumors. In this study, three cases with acute appendicitis symptoms are presented, including their clinical and histopathological findings. Microscopic examination detected GCCT in two cases and mixed adenoneuroendocrine carcinoma in one case, in addition to acute appendicitis.

Growth performance,nutrient digestibility,intestinal morphology,cecal mucosal cytokines and serum antioxidant responses of broiler chickens to dietary enzymatically treated yeast and coccidia challenge

Background There is a growing search for natural feed additives to alleviate the deleterious effects of coccidia infection in poultry production.This study aimed to investigate the effect of enzymatically treated yeast(ETY) on the growth performance,nutrient digestibility,intestinal morphology,antioxidative status,and cecal mucosa cytokines of coccidia-challenged broiler chickens.Methods From d 1 to 14 post hatching,480 broiler chickens were allocated to 3 corn-soybean meal-based experimental diets with increasing concentrations of ETY(0,1,or 2 g/kg).The experiment was designed as a randomized complete block design with body weight(BW) used as a blocking factor.On d 14 post hatching,the birds were re-randomized within each of the 3 experimental diets.Each of the 3 diet groups was split into a challenge or no-challenge group.This resulted in a 3 × 2 factorial arrangement of treatments.The coccidia challenge was administered on d 15 by an oral gavage.Results Dietary ETY improved(P < 0.05) the G:F of birds on d 21 regardless of the challenge state and linearly increased(P < 0.01) the apparent ileal digestibility of dry matter(DM),nitrogen,and gross energy(GE).The coccidia challenge decreased(P < 0.05) BW gain and feed intake of broiler chickens and reduced(P < 0.01) the total tract retention of DM,GE,and nitrogen.The coccidia challenge increased(P < 0.01) the mRNA gene expression of TNFα,IL-1β,IL-10,and IL-6 in the cecal mucosa.There was a tendency(P ne = 0.08) for ETY to linearly reduce IL-1β expression.Additionally,ETY supplementation increased(P < 0.05) the geexpression of OCLN.Serum catalase increased(P < 0.05) with dietary ETY in broiler chickens on d 21.Dietary ETY linearly increased(P < 0.05) the ileal villus height to crypt depth ratio,and ileal goblet cell density in broiler chickens.The ileal and excreta oocyst counts decreased(P < 0.01) with increasing supplementation of dietary ETY in coccidia-challenged broiler chickens on d 21.Conclusions Dietary ETY enhanced nutrient utilization and augmented intestinal development in broiler chickens.However,dietary ETY did not completely attenuate the adverse effects of a coccidia challenge in broiler chickens.

Autoimmune encephalitis after surgery for appendiceal cancer:A case report

BACKGROUND Primary cancer of the appendix is rare and often difficult to diagnose preoperatively due to the lack of specific clinical symptoms. Autoimmune encephalitis(AIE) is the most common cause of non-infectious encephalitis. The etiologies of AIE include tumors(paraneoplastic), infections(parainfections), or recessive infections. The tumors that have been reported to cause AIE include thymomas, ovarian teratomas, lung cancers, and breast cancers. However, there are no reports of AIE occurring after surgery for appendiceal cancer. This report describes the diagnosis and treatment of a patient with an appendiceal cancer and postoperative AIE.CASE SUMMARY We report the case of a 47-year-old man who was transferred to our hospital due to a recurrent low intestinal obstruction. Abdominal enhanced computed tomography was used to consider the possibility of a terminal ileal tumor with serous infiltration and lymph node metastasis. A right hemi-colectomy was performed under general anesthesia with an ileo-transcolon anastomosis and laparoscopic exploration. The postoperative pathologic evaluation revealed a high-grade goblet cell carcinoma of the appendix, accompanied by mesangial and abdominal lymph node metastases, and neural tube and vascular infiltration. The operation was completed without complication. The patient developed restlessness on postoperative day 4, and gradually developed a disturbance of consciousness on postoperative day 6. He was transferred to West China Hospital of Sichuan University and diagnosed with AIE.CONCLUSION Albeit rare, the occurrence of neurologic and psychiatric symptoms in patients with an appendiceal cancer postoperatively suggests the possibility of AIE.

Effect of dietary fiber and threonine content on intestinal barrier function in pigs challenged with either systemic E.coli lipopolysaccharide or enteric Salmonella Typhimurium

Background:The independent and interactive effects of dietary fiber(DF)and threonine(Thr)were investigated in growing pigs challenged with either systemic E.coli lipopolysaccharide(LPS)or enteric Salmonella Typhimurium(ST)to characterise their effect on intestinal barrier function.Results:In experiment 1,intestinal barrier function was assessed via oral lactulose and mannitol(L:M)gavage and fecal mucin analysis in pigs challenged with E.coli LPS and fed low fiber(LF)or high fiber(HF)diets with graded dietary Thr.Urinary lactulose recovery and L:M ratio increased(P<0.05)during the LPS inoculation period in LF fed pigs but not in HF fed pigs.Fecal mucin output was increased(P<0.05)in pigs fed HF compared to LF fed pigs.In experiment 2,RT-qPCR,ileal morphology,digesta volatile fatty acid(VFA)content,and fecal mucin output were measured in Salmonella Typhimurium challenged pigs,fed LF or HF diets with standard or supplemented dietary Thr.Salmonella inoculation increased(P<0.05)fecal mucin output compared to the unchallenged period.Supplemental Thr increased fecal mucin output in the HF-fed pigs(Fib×Thr;P<0.05).Feeding HF increased(P<0.05)VFA concentration in cecum and colon.No effect of either Thr or fiber on expression of gene markers was observed except a tendency(P=0.06)for increased MUC2 expression with the HF diet.Feeding HF increased goblet cell numbers(P<0.05).Conclusion:Dietary fiber appears to improve barrier function through increased mucin production capacity(i.e.,goblet cell numbers,MUC2 gene expression)and secretion(i.e.,fecal mucin output).The lack of effect of dietary Thr in Salmonella-challenged pigs provides further evidence that mucin secretion in the gut is conserved and,therefore,Thr may be limiting for growth under conditions of increased mucin production.

Goblet cell carcinoid of the appendix:Six case reports

BACKGROUND Goblet cell carcinoid(GCC)of the appendix is a rare tumor characterized by neuroendocrine and adenocarcinoma features.Accurate preoperative diagnosis is very difficult,with most patients complaining mainly of abdominal pain.Computed tomography shows swelling of the appendix,so diagnosis is usually made incidentally after appendectomy based on a preoperative diagnosis of appendicitis.Even if a patient undergoes preoperative colonoscopy,accurate endoscopic diagnosis is very difficult because GCC shows a submucosal growth pattern with invasion of the appendiceal wall.CASE SUMMARY Between 2017 and 2022,6 patients with GCC were treated in our hospital.The presenting complaint for 5 of these 6 patients was abdominal pain.All 5 patients underwent appendectomy,including 4 for a preoperative diagnosis of appendicitis and the other for diagnosis and treatment of an appendiceal tumor.The sixth patient presented with vomiting and underwent ileocecal resection for GCC diagnosed from preoperative biopsy.Although 2 patients with GCC underwent colonoscopy,no neoplastic changes were identified.Two of the six patients showed lymph node metastasis on pathological examination.As of the last followup(median:15 mo),all cases remained alive without recurrence.CONCLUSION As preoperative diagnosis of GCC is difficult,this possibility must be considered during surgical treatments for presumptive appendicitis.

Colon mucus in colorectal neoplasia and beyond

Little was known about mammalian colon mucus(CM)until the beginning of the 21^(st) century.Since that time considerable progress has been made in basic research addressing CM structure and functions.Human CM is formed by two distinct layers composed of gel-forming glycosylated mucins that are permanently secreted by goblet cells of the colonic epithelium.The inner layer is dense and impenetrable for bacteria,whereas the loose outer layer provides a habitat for abundant commensal microbiota.Mucus barrier integrity is essential for preventing bacterial contact with the mucosal epithelium and maintaining homeostasis in the gut,but it can be impaired by a variety of factors,including CM-damaging switch of commensal bacteria to mucin glycan consumption due to dietary fiber deficiency.It is proven that impairments in CM structure and function can lead to colonic barrier deterioration that opens direct bacterial access to the epithelium.Bacteria-induced damage dysregulates epithelial proliferation and causes mucosal inflammatory responses that may expand to the loosened CM and eventually result in severe disorders,including colitis and neoplastic growth.Recently described formation of bacterial biofilms within the inner CM layer was shown to be associated with both inflammation and cancer.Although obvious gaps in our knowledge of human CM remain,its importance for the pathogenesis of major colorectal diseases,comprising inflammatory bowel disease and colorectal cancer,is already recognized.Continuing progress in CM exploration is likely to result in the development of a range of new useful clinical applications addressing colorectal disease diagnosis,prevention and therapy.

GASTROINTESTINAL ABNORMALITIES IDENTIFIED BY FLUORESCENCE ENDOMICROSCOPY

Real-tine in vivo microscopic imaging has becomne a reality with the advent of confocal and nonlinear endomicroscopy.These devices are best utilized in conjunction with standard white light endoscopy.We evaluated the use of fuorescence endomicroscopy in detecting microscopic abnormalities in colonic tisues.Mice of C57bl/6 strain had intraperitoneal injection with azoxymethane once every week for five weeks and littermates,not exposed to azoxymethane served as controls.After 14 weeks,intestines were imaged by fuorescence endomicroscopy.The images show obvious cellular structural diferences between those two groups of mice.The difference in endomicroscopy imaging can be used for identifying tissues suspicious for neoplasia or other changes,leading to early diagnosis of gastrointestinal track of cancer.

Intestinal mucosal barrier in functional constipation:Dose it change?

BACKGROUND The intestinal mucosal barrier is the first line of defense against numerous harmful substances,and it contributes to the maintenance of intestinal homeostasis.Recent studies reported that structural and functional changes in the intestinal mucosal barrier were involved in the pathogenesis of several intestinal diseases.However,no study thoroughly evaluated this barrier in patients with functional constipation(FC).AIM To investigate the intestinal mucosal barrier in FC,including the mucus barrier,intercellular junctions,mucosal immunity and gut permeability.METHODS Forty FC patients who fulfilled the Rome IV criteria and 24 healthy controls were recruited in the Department of Gastroenterology of China-Japan Friendship Hospital.The colonic mucus barrier,intercellular junctions in the colonic epithelium,mucosal immune state and gut permeability in FC patients were comprehensively examined.Goblet cells were stained with Alcian Blue/Periodic acid Schiff(AB/PAS)and counted.The ultrastructure of intercellular junctional complexes was observed under an electron microscope.Occludin and zonula occludens-1(ZO-1)in the colonic mucosa were located and quantified using immunohistochemistry and quantitative real-time polymerase chain reaction.Colonic CD3+intraepithelial lymphocytes(IELs)and CD3+lymphocytes in the lamina propria were identified and counted using immunofluorescence.The serum levels of D-lactic acid and zonulin were detected using enzyme-linked immunosorbent assay.RESULTS Compared to healthy controls,the staining of mucus secreted by goblet cells was darker in FC patients,and the number of goblet cells per upper crypt in the colonic mucosa was significantly increased in FC patients(control,18.67±2.99;FC,22.42±4.09;P=0.001).The intercellular junctional complexes in the colonic epithelium were integral in FC patients.The distribution of mucosal occludin and ZO-1 was not altered in FC patients.No significant differences were found in occludin(control,5.76E-2±1.62E-2;FC,5.17E-2±1.80E-2;P=0.240)and ZO-1(control,2.29E-2±0.93E-2;FC,2.68E-2±1.60E-2;P=0.333)protein expression between the two groups.The mRNA levels in occludin and ZO-1 were not modified in FC patients compared to healthy controls(P=0.145,P=0.451,respectively).No significant differences were observed in the number of CD3+IELs per 100 epithelial cells(control,5.62±2.06;FC,4.50±2.16;P=0.070)and CD3+lamina propria lymphocytes(control,19.69±6.04/mm^(2);FC,22.70±11.38/mm^(2);P=0.273).There were no significant differences in serum D-lactic acid[control,5.21(4.46,5.49)mmol/L;FC,4.63(4.31,5.42)mmol/L;P=0.112]or zonulin[control,1.36(0.53,2.15)ng/mL;FC,0.94(0.47,1.56)ng/mL;P=0.185]levels between FC patients and healthy controls.CONCLUSION The intestinal mucosal barrier in FC patients exhibits a compensatory increase in goblet cells and integral intercellular junctions without activation of mucosal immunity or increased gut permeability.

Dietary iron regulates intestinal goblet cell function and alleviates Salmonella typhimurium invasion in mice

Iron is an important micronutrient that plays a vital role in host defenses and bacterial pathogenicity. As iron treatments increase the risk of infection by stimulating the growth and virulence of bacterial pathogens, their roles in anti-infection immunity have frequently been underestimated. To estimate whether adequate dietary iron intake would help defend against pathogenic bacterial infection, mice were fed iron-deficient(2 mg kg-1feed), iron-sufficient(35 mg kg-1feed), or iron-enriched diet(350 mg kg-1feed) for 12 weeks, followed by oral infection with Salmonella typhimurium. Our results revealed that dietary iron intake improved mucus layer function and decelerated the invasion of the pathogenic bacteria, Salmonella typhimurium. Positive correlations between serum iron and the number of goblet cells and mucin2 were found in response to total iron intake in mice.Unabsorbed iron in the intestinal tract affected the gut microbiota composition, and the abundance of Bacteroidales, family Muribaculaceae, was positively correlated with their mucin2 expression. However, the results from antibiotic-treated mice showed that the dietary iron-regulated mucin layer function was not microbial-dependent. Furthermore, in vitro studies revealed that ferric citrate directly induced mucin2 expression and promoted the proliferation of goblet cells in both ileal and colonic organoids. Thus, dietary iron intake improves serum iron levels, regulates goblet cell regeneration and mucin layer function, and plays a positive role in the prevention of pathogenic bacteria.

High amylose to amylopectin ratios in nitrogen-free diets decrease the ileal endogenous amino acid losses of broiler chickens

This study explored the variation of ileal endogenous amino acid(IEAA)losses and its influencing factors in chickens offered nitrogen-free diets(NFD)containing different ratios of amylose to amylopectin(AM/AP).A total of 252 broiler chickens at 28 d old were randomly allocated into 7 treatment groups for a 3-d trial.The dietary treatments included a basal diet(control),a NFD containing corn starch(CS),and 5NFD with AM/AP ratios of 0.20,0.40,0.60,0.80,and 1.00,respectively.As the AM/AP ratio increased,the IEAA losses of all AAs,starch digestibility and maltase activity linearly decreased(P<0.05),but the DM digestibility linearly and quadratically decreased(P<0.05).Compared with the control,the NFD increased the number of goblet cells and its regulatory genes mucin-2 and krüppel-like factor 4(KLF-4)while decreasing serum glucagon and thyroxine concentrations,ileal villus height,and crypt depth(P<0.05).Additionally,NFD with lower AM/AP ratios(0.20 and 0.40)decreased the ileal microbiota species richness(P<0.05).In all NFD groups,the number of Proteobacteria increased whereas the abundance of Firmicutes dropped(P<0.05).However,the broilers in the AM/AP 0.60 group were closer to the digestive physiological state of chickens fed the control diet,with no significant change in maltase activity and mucin-2 expression(P<0.05).In conclusion,increasing AM/AP ratio in a NFD decreased the IEAA losses and the apparent ileal digestibility of starch but inevitably resulted in malnutrition and disruption of gut microbiota homeostasis.This study recommends AM/AP in NFD at 0.60 to measure IEAA of broiler chickens.

Poly-β-hydroxybutyrate alleviated diarrhea and colitis via Lactobacillus johnsonii biofilm-mediated maturation of sulfomucin

Maintainance of sulfomucin is a key end point in the treatment of diarrhea and inflammatory bowel disease(IBD).However,the mechanisms underlying the microbial sense to sulfomucin are poorly understood,and to date,there are no therapies targeting the secretion and maturation of sulfomucin in IBD.Herein,we biosynthesized poly-β-hydroxybutyrate(PHB)and found that PHB could alleviate inflammation caused by diarrhea and colitis by enhancing the differentiation of sulfomucin.Microbiota transplantation and clearance together demonstrate that PHB promoting sulfomucin is mediated by Lactobacillus johnsonii(L.johnsonii).Further studies revealed that PHB provides a favorable niche for L.johnsonii biofilm formation to resist disturbance and support its growth.L.johnsonii-biofilm alleviates colitis by regulating fucose residues to promote goblet cell differentiation and subsequent sulfomucin maturation.Importantly,PHB alleviates colitis by enhancing sulfomucin secretion and maturation in a L.johnsonii-dependent manner.PHB represents a class of guardians,acting as a safe probiotic-biofilm delivery system that significantly promotes probiotic proliferation.Altogether,this study adds weight to the possible role of probiotics and functional materials in the treatment of intestinal inflammation.The application of PHB and biofilm self-coating L.johnsonii carries high translational potential and may be of clinical relevance.

Endogenous mucin conveyed to the mucosa with microbes can assure lumen fermentation and large intestinal security-swine versus fowl

Endogenous protein leaving the ileum largely consists of accrued mucins from the upper gastrointestinal tract(GIT)that had resisted digestion.The amounts released rely on their mucosal generation during enteral feeding which vary with age as well as diet.These digestion resistant proteins of endogenous origin continue to be unavailable in the large intestine,whereas those of dietary origin provide amino acids that largely support the existing microbial population while denying limited amounts for absorption.Other mucins pre-exist within the large intestine as two layers at the lumen surface.A loose layer harboring a diverse microbial population is superimposed on the unstirred water layer(USWL)which simultaneously acts as an obstacle to microbes at the loose layer while performing as a molecular sieve for nutrients.The USWL is formed through interplay between enterocyte and goblet cells;however,the basis for presence of the loose layer is elusive.Large intestinal fermentation predominates within the colon of swine,whereas fowl employ their ceca.Motility within the colon of swine segregates fine materials into haustrae out-pocketings that parallel their placement within the ceca of fowl.Viscous mucins from small intestinal endogenous losses may envelop microbes within the large intestinal lumen to present successive adherents on the USWL that assemble its loose layer.The loose layer continually functions as a microbial reservoir in support of lumen fermentation.Microbial catabolism of mucin within the loose layer is known to be slow,but its proximity to the enterocyte is of advantage to enterocyte absorption with by-product amino acids fostering the USWL.

TRIM34 attenuates colon inflammation and tumorigenesis by sustaining barrier integrity

Loss of the colonic inner mucus layer leads to spontaneously severe colitis and colorectal cancer.However,key host factors that may control the generation of the inner mucus layer are rarely reported.Here,we identify a novel function of TRIM34 in goblet cells(GCs)in controlling inner mucus layer generation.Upon DSS treatment,TRIM34 deficiency led to a reduction in Muc2 secretion by GCs and subsequent defects in the inner mucus layer.This outcome rendered TRIM34-deficient mice more susceptible to DSS-induced colitis and colitis-associated colorectal cancer.Mechanistic experiments demonstrated that TRIM34 controlled TLR signaling-induced Nox/Duox-dependent ROS synthesis,thereby promoting the compound exocytosis of Muc2 by colonic GCs that were exposed to bacterial TLR ligands.Clinical analysis revealed that TRIM34 levels in patient samples were correlated with the outcome of ulcerative colitis(UC)and the prognosis of rectal adenocarcinoma.This study indicates that TRIM34 expression in GCs plays an essential role in generating the inner mucus layer and preventing excessive colon inflammation and tumorigenesis.

PIM1-HDAC2 axis modulates intestinal homeostasis through epigenetic modification

The mucosal barrier is crucial for intestinal homeostasis,and goblet cells are essential for maintaining the mucosal barrier integrity.The proviral integration site for Moloney murine leukemia virus-1(PIM1)kinase regulates multiple cellular functions,but its role in intestinal homeostasis during colitis is unknown.Here,we demonstrate that PIM1 is prominently elevated in the colonic epithelia of both ulcerative colitis patients and murine models,in the presence of intestinal microbiota.Epithelial PIM1 leads to decreased goblet cells,thus impairing resistance to colitis and colitis-associated colorectal cancer(CAC)in mice.Mechanistically,PIM1 modulates goblet cell differentiation through the Wnt and Notch signaling pathways.Interestingly,PIM1 interacts with histone deacetylase 2(HDAC2)and downregulates its level via phosphorylation,thereby altering the epigenetic profiles of Wnt signaling pathway genes.Collectively,these findings investigate the unknown function of the PIM1-HDAC2 axis in goblet cell differentiation and ulcerative colitis/CAC pathogenesis,which points to the potential for PIM1-targeted therapies of ulcerative colitis and CAC.