Cytokine tumor necrosis factor alpha induces intestinal epithelial barrier dysfunction.

BACKGROUND AND AIMS: Epithelial barrier dysfunction is involved in a number of diseases in the body. The mechanism is to be further understood. The present study aimed to investigate the role of one of the common microbial products, flagellin (FGN), in the induction of intestinal epithelial barrier dysfunction. METHODS: We collected the colon epithelium specimens from 40 patients with ulcerative colitis (UC), 40 patients with Crohn's disease (CD) and 40 healthy volunteers. The expression of toll like receptors (TLR)5 of the specimens was assessed by RT-PCR and western blotting. The expression of tumor necrosis factor alpha (TNFα) and its role in compromising the barrier function in the intestinal epithelial cells, T84 cells, were observed by a cell culture model. RESULTS: The results showed that the expression of TLR5 was observed in the colon epithelium of healthy subjects that was increased in UC patients and further increased in CD patients. Treating T84 cells with FGN increased the expression of TNFα in the cells that caused the T84 cell apoptosis as well as compromised the T84 monolayer barrier function, which could be prevented by knocking down the gene of TNFα in T84 cells. CONCLUSIONS: We conclude that the human colon epithelial cells express detectable TLR5 that is increased in patients with CD and UC. The exposure to FGN can increase the expression of TNFα that further compromises the intestinal epithelial barrier function.

A fatal outcome in a patient with coeliac disease who suspended a strict gluten-free diet: a case report.

Coeliac disease (CD) is an autoimmune small bowel disease that occurs in susceptible individuals that develop an immunological reaction to gluten. A strict gluten-free diet (GFD) is the primary treatment for CD. This case report describes a patient with CD recurrence due to a discontinuation of a strict GFD by the patient. After recurrence, the patient developed fever and pancytopaenia, and quickly died of haemophagocytic lymphohistiocytosis (HLH). To the best of our knowledge, this is the first description of a case of CD associated with HLH due to discontinued GFD, which may contribute to improving the awareness of the importance of maintaining a strict GFD and having regular follow-up examinations.

Mesenteric phlebosclerosis with amyloidosis in association with the long-term use of medicinal liquor: A case report.

BACKGROUND: Mesenteric phlebosclerosis (MP) is a rare disease of the colon. The clinical manifestations of this disease are nonspecific and it may easily be misdiagnosed. We report a case of MP with amyloidosis in the colonic vessel walls in a patient with hypertension who had been consuming Chinese medicinal liquor for 10 years. We also review the relevant literature and summarize the characteristics of MP in patients in mainland China. CASE SUMMARY: A 64-year-old man was referred to our department from his primary hospital because of abdominal pain, diarrhea, and fever for almost 10 d. Computed tomography showed colon wall thickening, with threadlike calcifications in the mesenteric vein in the transverse colon. Colonoscopy revealed purple-blue mucosa with multiple ulcers in the ascending and transverse colon. Biopsy showed thickening and calcification of the vein walls, perivascular and mucosal collagen degeneration, and amyloidosis. The patient had been consuming Chinese medicinal liquor, mainly that made from gardenia fruit, for 10 years. Based on these results, a diagnosis of MP with amyloidosis was made. After conservative treatment, the patient's discomfort subsided and he was followed closely. The use of Chinese herbal medicine was suspected to play a role in the pathogenesis of MP. CONCLUSION: The clinical manifestations of MP are nonspecific. Recognition of its typical imaging findings, including multiple calcifications on computed tomography and purple-blue mucosal discoloration on colonoscopy, is vital.

A stubborn anemia caused by ectopic pancreas bleeding in the jejunum revealed by capsule endoscopy.

Ectopic pancreas is extremely rare in clinical setting. Meanwhile, a stubborn anemia without obvious dark bloody stool due to ectopic pancreas diagnosed by capsule endoscopy has not been reported. We reported a case of an ectopic pancreas inducing obscure gastrointestinal bleeding in a 70-year-old woman presenting as stubborn anemia, which was diagnosed by capsule endoscopy. The patient recovered well after resection the lesion. Diagnosis of ectopic pancreas is extremely difficult with conventional techniques. Endoscopists should pay more attention to the ectopic pancreas as a rare differential consideration for occult intestinal bleeding.

Segmental small bowel necrosis associated with antiphospholipid syndrome: a case report.

Antiphospholipid syndrome is a multi-system disease characterized by the formation of thromboembolic complications and/or pregnancy morbidity, and with persistently increased titers of antiphospholipid antibodies. We report the case of a 50-year-old, previously healthy man who presented with fever and new-onset, dull abdominal pain. A contrast-enhanced computed tomography scan showed segmental small bowel obstruction, for which an emergency laparotomy was performed. Histopathologic examination of resected tissues revealed multiple intestinal and mesenteric thromboses of small vessels. Laboratory tests for serum antiphospholipid (anticardiolipin IgM) and anti-ß2-glycoprotein I antibodies were positive. Despite proactive implementation of anticoagulation, steroid, and antibiotic therapies, the patient's condition rapidly deteriorated, and he died 22 d after admission. This case highlights that antiphospholipid syndrome should be suspected in patients with unexplainable ischemic bowel and intestinal necrosis presenting with insidious clinical features that may be secondary to the disease, as early diagnosis is critical to implement timely treatments in order to ameliorate the disease course.

Extranodal NK/T-cell lymphoma, nasal type: a case report of 7-year natural course and review of literature.

We present our experience a vary case of Extranodal natural killer (NK)/T-cell lymphomas, nasal type (ENKL) who survived for 7 years, and review the recent advances on the differential diagnosis.

Centralized isolation of Helicobacter pylori from multiple centers and transport condition influences.

AIM: To evaluate the efficacy of centralized culture and possible influencing factors. METHODS: From January 2010 to July 2012, 66452 patients with suspected Helicobacter pylori (H. pylori) infection from 26 hospitals in Zhejiang and Jiangsu Provinces in China underwent gastrointestinal endoscopy. Gastric mucosal biopsies were taken from the antrum for culture. These biopsies were transported under natural environmental temperature to the central laboratory in Hangzhou city and divided into three groups based on their transport time: 5, 24 and 48 h. The culture results were reported after 72 h and the positive culture rates were analyzed by a χ (2) test. An additional 5736 biopsies from H. pylori-positive patients (5646 rapid urease test-positive and 90 (14)C-urease breath test-positive) were also cultured for quality control in the central laboratory setting. RESULTS: The positive culture rate was 31.66% (21036/66452) for the patient samples and 71.72% (4114/5736) for the H. pylori-positive quality control specimens. In the 5 h transport group, the positive culture rate was 30.99% (3865/12471), and 32.84% (14960/45553) in the 24 h transport group. In contrast, the positive culture rate declined significantly in the 48 h transport group (26.25%; P < 0.001). During transportation, the average natural temperature increased from 4.67 to 29.14 °C, while the positive culture rate declined from 36.67% (1462/3987) to 24.12% (1799/7459). When the temperature exceeded 24 °C, the positive culture rate decreased significantly, especially in the 48 h transport group (23.17%). CONCLUSION: Transportation of specimens within 24 h and below 24 °C is reasonable and acceptable for centralized culture of multicenter H. pylori samples.

Competitive inhibition of adherence of enterotoxigenic Escherichia coli, enteropathogenic Escherichia coli and Clostridium difficile to intestinal epithelial cell line Lovo by purified adhesin of Bifidobacterium adolescentis 1027.

AIM: To observe competitive inhibition of adherence of enterotoxigenic Escherichia coli (ETEC), enteropathogenic Escherichia coli (EPEC) and Clostridium difficile (C. difficile) to intestinal epithelial cell line Lovo by purified adhesin of Bifidobacterium adolescentis 1027 (B. ado 1027). METHODS: The binding of bacteria to intestinal epithelial cell line Lovo was counted by adhesion assay. The inhibition of adherence of ETEC, EPEC and C. difficile to intestinal epithelial cell line Lovo by purified adhesin of B. ado 1027 was evaluated quantitatively by flow cytometry. RESULTS: The purified adhesin at the concentration of 10 microg/mL, 20 microg/mL and 30 microg/mL except at 1 microg/mL and 5 microg/mL could inhibit significantly the adhesion of ETEC, EPEC and C. difficile to intestinal epithelial cell line Lovo. Moreover, we observed that a reduction in bacterial adhesion was occurred with increase in the concentration of adhesin, and MFI (Mean fluorescent intensity) was decreased with increase in the concentration of adhesin. CONCLUSION: The purified adhesin of B. ado 1027 can inhibit the adhesion of ETEC, EPEC and C. difficile to intestinal epithelial cell line Lovo in a dose-dependent manner.

Contribution of eIF-4E inhibition to the expression and activity of heparanase in human colon adenocarcinoma cell line: LS-174T.

AIM: Heparanase degrades heparan sulfate proteoglycans (HSPGs) and is a critical mediator of tumor metastasis and angiogenesis. Recently, it has been cloned as a single gene family and found to be a potential target for antimetastasis drugs. However, the molecular basis for the regulation of heparanase expression is still not quite clear. The aim of this study was to determine whether the expression of eukaryotic initiation factor 4E (eIF-4E) correlated with the heparanase level in tumor cells and to explore the correlation between heparanase expression and metastatic potential of LS-174T cells. METHODS: A 20-mer antisense s-oligodeoxynucleotide (asODN) targeted against the translation start site of eIF-4E mRNA was introduced into LS-174T cells by lipid-mediated DNA-transfection. eIF-4E protein and mRNA levels were detected by Western blot analysis and RT-PCR, respectively. Heparanase activity was defined as the ability to degrade high molecular weight (40-100 kDa) radiolabeled HS (heparan sulfate) substrate into low molecular weight (5-15 kDa) HS fragments that could be differentiated by gel filtration chromatography. The invasive potential of tumor cell in vitro was observed by using a Matrigel invasion assay system. RESULTS: The 20-mer asODN against eIF-4E specifically and significantly inhibited eIF-4E expression at both transcriptional and translational levels. As a result, the expression and activity of heparanase were effectively retarded and the decreased activity of heparanase resulted in the decreased invasive potential of LS-174T. CONCLUSION: eIF-4E is involved in the regulation of heparanase production in colon adenocarcinoma cell line LS-174T, and its critical function makes it a particularly interesting target for heparanase regulation. This targeting strategy in antisense chemistry may have practical applications in experimental or clinical anti-metastatic gene therapy of human colorectal carcinoma.

Effect of protein kinase C on multidrug resistance of multidrug-resistant colorectal cancer LoVo/Adr cells.

OBJECTIVE: To observe the effect of protein kinase C (PKC) on the multidrug resistance of multidrug-resistant colorectal cancer LoVo/Adr cells and explore the mechanism. METHODS: The changes of PKC activity in LoVo/Adr cells in response to treatment with staurosporine (SP) and phorbol-12-myristate-13-acetate (PMA) were detected by way of 32P incorporation. The effect of PKC on adriamycin uptake in LoVo/Adr cells was detected by flow cytometry. Reverse transcriptase-PCR was utilized to observe the effect of PKC on mdr1 gene expression. RESULTS: PMA evinced bi-directional regulation of PKC activity in LoVo/Adr cells, and SP significantly inhibited membrane and cytosol fraction of PKC activity. Preincubation with PMA for 30 min caused the uptake of adriamycin to decrease significantly, but when the preincubation was prolonged to 24 h, significant increase occurred in adriamycin uptake. Neither PMA nor SP, however, could affect the expression of mdr1 gene. CONCLUSION: PKC regulates multidrug resistance of the cells through mechanisms other than regulation of mRNA level of mdr1 gene.

Establishment of dextran sulfate sodium-induced ulcerative colitis model in mice.

OBJECTIVE: To establish a model of ulcerative colitis in mice. METHODS: The mice were given 5% dextran sulfate sodium (DSS) solution freely for 7 consecutive days after which distilled water was given in stead for another 10 d, to complete one cycle of whole treatment plan that consisted of 4 such cycles. The symptoms were observed daily. At the end of the first and the fourth cycle respectively, the mice were killed for examining the whole colon under anatomic microscopy and serial tissue sections were prepared for histological observation. RESULTS: The symptoms observed in the mice including hematochezia diarrhea and loss of the body weight were similar to those of ulcerative colitis patients. Histological examination revealed infiltration of neutrocytophilia and lymphocythemia, with loss of integrity of the colon mucosa in the gland. CONCLUSION: This mouse model of ulcerative colitis can be easily induced and readily applied in various studies of ulcerative colitis.

[Effects of catalase on the mitochondria and apoptosis of SW480 cells].

OBJECTIVE: To investigate the effects of catalase on the mitochondria and apoptotic behavior of SW480 cells treated by lipopolysaccharide (LPS), and the changes in intracellular expression of nuclear factor kappaB (NF-kappaB). METHODS: LPS-stimulated SW480 cells were treated with catalase and the changes in their mitochondria and apoptotic behavior were observed by transmission electron microscopy. The expression of NF-kappaB was detected by immunohistochemical method. RESULTS: Mitochondria damages and apoptosis were slightly diminished in the LPS-stimulated cells with catalase pretreatment, and the NF-kappaB expression was also decreased. CONCLUSION: Catalase protect the mitochondria of SW480 cells from damages by LPS and inhibit the cell apoptosis, possibly due to the activation of NF-kappaB.

[Expression of pro-inflammation cytokines and activation of nuclear factor kappab in the intestinal mucosa of mice with ulcerative colitis].

OBJECTIVE: To investigate the expression of pro-inflammation cytokines and activation of nuclear factor kappaB (NF-kappaB) in mouse models of ulcerative colitis. METHODS: Mouse models of ulcerative colitis were established by oral administration of 5% dextran sulfate sodium for 7 d, and the expression of tumor necrosis factor (TNF)- alpha and interleukin (IL)-1beta in the intestinal mucosa were detected by semi-quantitative reverse transcriptional (RT) PCR. The activation of NF-kappaB in the intestinal mucosa was evaluated by electrophoretic mobility shift assay (EMSA). RESULTS: The expressions of TNF-alpha and IL-1beta were increased in the intestinal mucosa (P=0.009), and the nuclear binding activity of NF-kappaB was also up-regulated after the onset of colitis. CONCLUSION: Pro-inflammatory cytokines play important roles in the pathogenesis of UC, and may exacerbate the inflammation of the intestinal mocosa and cause apoptosis of the epithelial cells, possibly under the regulation of NF-kappaB activation.

[Role of Helicobacter pylori infection in the pathogenesis and clinical outcome of childhood acute idiopathic thrombocytopenic purpura].

OBJECTIVE: To investigate the role of Helicobacter pylori (Hp) and its products cytotoxin-associated protein (Cag A), vacuolating cytotoxin (VacA) in childhood acute idiopathic thrombocytopenic purpura (aITP), to evaluate the effect of Hp on their clinical outcome. METHODS: Subjects were enrolled according to case-control design, including 184 aITP children and 154 healthy controls. They were inquired for demographic characteristics, the risk factors regarding Hp infection and ITP through a uniformed questionnaire. Patients with Hp infection were diagnosed by combined detection of serum Hp antibodies and stool antigens. CagA and VacA proteins were tested by ELISA method. In addition, clinical data and follow-up data of aITP children were collected. Non-conditional logistic regression and t test were applied for statistical analysis. RESULTS: (1) The prevalence of Hp infection in aITP children and controls were 41.30% and 35.71%, respectively. No association between Hp infection and children aITP was found with OR of 1.170 (95%CI: 0.7163 - 1.673) after adjusting for confounding variables. (2) No statistical differences regarding initial platelet counts, megakaryocytes counts and the constituent ratio were found between the aITP children with and without Hp infection (P > 0.05). (3) No differences regarding initial platelet counts were found between aITP children with and without the expression of CagA (P > 0.05). The follow-up data showed that 32.88% of aITP children with Hp infection, as well as 29.70% of aITP children without Hp infection developed into cITP. No association between Hp infection and development to cITP was found with adjusted OR 1.171 (95%CI: 0.555 - 2.11 2). CONCLUSIONS: The results didn't suggest that Hp is unlikely to play a role in the onset of childhood aITP, and in the development of aITP to cITP.

[Role of eukaryotic initiation factor-4E (eIF-4E) in regulation of expression of NF-kappaB and its subsequent influence on transcription and activity of heparanase in human colon adenocarcinoma cell line].

BACKGROUND & OBJECTIVE: Recent studies have revealed that some transcription factors may be translationally regulated by eukaryotic initiation factor-4E (eIF-4E) in human cancer. These modifications in the expression levels of the key transcription factors will alter the expression of some malignancy-related gene products at the transcriptional levels. The current study was designed to investigate the effect of eIF-4E on the expression and activity of NF-kappaB and observe how the activity level of NF-kappaB contributes, as a secondary effect, to the transcription of heparanase in human colon adenocarcinoma LS-174T cells. METHODS: In order to repress the expression of eIF-4E, a 20-mer antisense oligodeoxynucleotide (ASODN) targeted against the translation start site of eIF-4E mRNA was transfected into human colorectal cancer cell line LS-174T via liposome reagent, followed by assessment of the activity and the protein expression of NF-kappaB by an electrophoretic gel mobility shift assay (EMSA) and Western blot analysis respectively. The alterations of heparanase expression were examined by RT-PCR and Western blot analysis, and heparanase activity in LS-174T cells was measured by specific enzymatic activity test using radiolabeled heparan sulfate as the substrate and gel filtration chromatography for the analysis of the degradation product. RESULTS: The 20-mer ASODN against eIF-4E specifically and significantly inhibited eIF-4E expression at both transcriptional and translational levels, and the repression of eIF-4E gene expression was correlated with decreased expression levels and activity of NF-kappaB protein. Furthermore, this down regulation of the ubiquitous transcription factor NF-kappaB led to reduced transcription of the heparanase gene, and the transfected cells also showed a considerable decrease in heparanase protein and activity. CONCLUSION: These results suggest that eIF-4E play an important role in translational regulation of NF-kappaB expression in LS-174T cells.NF-kappaB is an essential factor in the regulatory mechanisms of heparanase gene transcription, and the suppressed NF-kappaB activity in LS-174T cells may significantly reduce the transcriptional expression of heparanase that consequently leads to decreased heparanase protein expression and enzymatic activity.