OBJECTIVE: To investigate the distribution of pulmonary surfactant protein A (SP-A) like molecules and the bridge of frontier host defense and adaptive immune response cell of CD68 positive macrophages in inflammatory bowel disease (IBD). METHODS: Surgical specimens derived from involved areas and normal area of the colon with Crohn disease (CD) and ulcerative colitis (UC) were obtained from Department of Pathology, Rhode Island Hospital, Brown University Medical Center. The distribution of SP-A like molecule in intestine of IBD was detected by immunohistochemistry. RESULTS: SP-A like molecule located in epithelia of intestine, the surface of intestine villi, blood vessels of connective tissue, and some inflammatory cells. The number of macrophages with both SP-A like molecule and CD68 positive was dramatically increased in the inflammatory area than the normal area. Some CD68 positive macrophages expressed SP-A like immunoreactivity by immunofluorescence double labeling. CONCLUSION SP-A is an important host defense molecule in lung, and SP-A expression in large intestine may reflect a close relation between 2 organs in immune response towards inflammation.
Antigens, CD ; metabolism ; Antigens, Differentiation, Myelomonocytic ; metabolism ; Colitis, Ulcerative ; immunology ; metabolism ; Colon ; metabolism ; Crohn Disease ; immunology ; metabolism ; Humans ; Immunohistochemistry ; Inflammatory Bowel Diseases ; immunology ; metabolism ; Macrophages ; immunology ; metabolism ; Pulmonary Surfactant-Associated Proteins ; genetics ; metabolism

BACKGROUND/AIMS: This animal study aimed to define the underlying cellular mechanisms of intestinal barrier dysfunction. METHODS: Rats were fed 4% with dextran sodium sulfate (DSS) to induce experimental colitis. We analyzed the sugars in 24-hour urine output by high pressure liquid chromatography. The expression of claudins, mannan-binding lectin (MBL), and MBL-associated serine proteases 2 (MASP-2) were detected in the colonic mucosa by immunohistochemistry; and apoptotic cells in the colonic epithelium were detected by the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling method assay. RESULTS: The lactulose and sucralose excretion levels in the urine of rats with DSS-induced colitis were significantly higher than those in the control rats. Mannitol excretion was lower and lactulose/mannitol ratios and sucralose/mannitol ratios were significantly increased compared with those in the control group (p<0.05). Compared with the controls, the expression of sealing claudins (claudin 3, claudin 5, and claudin 8) was significantly decreased, but that of claudin 1 was increased. The expression of pore-forming claudin 2 was upregulated and claudin 7 was downregulated in DSS-induced colitis. The epithelial apoptotic ratio was 2.8%+/-1.2% in controls and was significantly increased to 7.2%+/-1.2% in DSS-induced colitis. The expression of MBL and MASP-2 in the intestinal mucosa showed intense staining in controls, whereas there was weak staining in the rats with colitis. CONCLUSIONS: There was increased intestinal permeability in DSS-induced colitis. Changes in the expression and distribution of claudins, increased epithelial apoptosis, and the MASP-2-induced immune response impaired the intestinal epithelium and contributed to high intestinal permeability.
Animals ; Apoptosis/*physiology ; Claudins/*metabolism ; Colitis/chemically induced/immunology/*physiopathology ; Colon/immunology/physiopathology ; Dextran Sulfate ; Intestinal Mucosa/*physiopathology ; Lactulose/metabolism ; Mannitol/metabolism ; Mannose-Binding Lectin/*immunology ; Permeability ; Rats ; Rats, Sprague-Dawley ; Sucrose/analogs & derivatives/metabolism ; Up-Regulation

OBJECTIVETo investigate the influence of Weichang Anwan on the treatment of IBS-D in model rats.

METHODAnimal model of compound diarrhea was induced by a lactose enriched diet in the Wistar rat, combining with restraint stress. At first, the best cycle of taking medicine was tested. In order to decide the best cycle of taking medicine, 24 female Wistar rats were randomly divided into normal control group, model group and 60 mg x kg(1) x d(-1) weichangan group. The rate of weight increase, the rate of diarrhea, the incubation period of diarrhea and the diarrhea index were observed. And then 45 female Wistar rats randomly divided into five groups: normal control group, model group and Weichang Anwan groups of high, medium and low doses( 80, 60, 40 mg x kg(-1) x d(-1)). The mast cells in mucous membrane were observed by light microscope. The level of NO in blood serum was checked by the method of nitrate reductase. 5-HT in blood serum was detected by fluorimetry. The level of SP in colon was measured by radioimmunoassay.

RESULTAfter taking Weichang Anwan for 4 days, the rate of weight increase in Weichangan group was higher than the model group's. And the rate of diarrhea was lower significantly. So the best cycle of taking medicine was 4 days. The levels of NO and 5-HT in blood serum decreased remarkably in the model group than those of the normal control group. At the same time, the amount of the mast cells and the level of SP in colon significantly increased. Compared with the model group, the levels of NO and 5-HT in blood serum increased remarkably in the groups of high doses and medium doses. Meanwhile, the amount of the mast cells and the level of SP in colon decreased significantly.

CONCLUSIONWeichang Anwan has the effect of antidiarrhea. It can adjust the levels of NO and 5-HT in blood serum and can inhibit the expression of SP in colon which can active the mast cell. Weichangan can also decrease the amount of the mast cells directly.

Animals ; Colon ; drug effects ; immunology ; metabolism ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Humans ; Irritable Bowel Syndrome ; drug therapy ; immunology ; metabolism ; Mast Cells ; immunology ; Nitric Oxide ; blood ; Random Allocation ; Rats ; Rats, Wistar ; Serotonin ; blood ; Substance P ; metabolism

OBJECTIVETo explore the effects of Huoxiang Zhengqi liquid (HXZQ) on enteric mucosal immune responses in mice with Bacillus dysenteriae and Salmonella typhimurium induced diarrhea (BSD).

METHODMice were randomly divided into four groups with 10 mice in each group: control group (control), BSD group, Huoxiang Zhengqi liquid treated BSD groups at high dosage and low dosage (HXZQ high, HXZQ low). HXZQ was administrated from the day of diarrhea induction at dosage of 5.21 g kg(-1) and 0.52 g kg (-1) respectively. Peyer's patch and periphery lymphocytes were prepared for flow cytometry, and level of TNF-alpha in periphery and enteric tissue homogenate were determined with ELISA. Student's t-test was used for statistics.

RESULTMice in BSD group started showing continuous diarrhea at the day of induction till the fourth day when the mice were sacrificed. Diarrhea in the mice of HXZQ high and low groups lasted for 36 and 54 h respectively. There were more CD4+ and CD8+ cells in periphery, less CD4+ cells in peyer's patch in BSD mice comparing to normal mice. In peyer's patch, there were more CD8+ cells in mice in HXZQ high and low groups and more CD4+ in mice in HXZQ high group. Higher level TNF-alpha in periphery and intestinal tissue homogenate in BSD group were observed. Mice in HXZQ high group showed the decreased level TNF-alpha in periphery and enteric tissue homogenate.

CONCLUSIONThe immune regulation on peyer's patch CD4+ and CD8+ cells and suppression on TNF-alpha level in enteric homogenate might partially explain the effect of HXZQ on improvement of BSD.

Animals ; CD4-CD8 Ratio ; Colon ; immunology ; metabolism ; pathology ; Diarrhea ; immunology ; metabolism ; microbiology ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Dysentery, Bacillary ; immunology ; metabolism ; microbiology ; Immunity, Mucosal ; drug effects ; Intestinal Mucosa ; immunology ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Peyer's Patches ; drug effects ; immunology ; pathology ; Plants, Medicinal ; chemistry ; Random Allocation ; Salmonella Infections ; immunology ; metabolism ; microbiology ; Salmonella typhimurium ; immunology ; Shigella dysenteriae ; immunology ; T-Lymphocyte Subsets ; drug effects ; immunology ; pathology ; Tumor Necrosis Factor-alpha ; blood ; metabolism

UNLABELLEDTo provide a reliable animal model for study of human CMV disease in gastrointestinal track, we tried to infect with murine cytomegalovirus (MCMV) in mice that were received allogenetic skin transplantation under immunosuppression. (1) Skin transplantation was performed between 18 donor C57BL/6 mice and 72 recipient BALB/c mice. (2) All recipient mice were then given Cyclosporine at 12 mg/kg daily for 2 weeks by intraperitoneal injection. Mice were randomly divided into 3 groups. Two experimental groups were received MCMV-infected mouse embryonic fibroblasts (MEF) at 10(4) PFU and 10(5) PFU respectively, and the control group received MEF only. We observed any possibly pathophysiological behavior changes and recorded the changes in body weight. The mice were sacrificed at 5d, 9d, 14d, 21d post infection and colon tissue was collected for analysis.

RESULTSMice infected with MCMV at 10(5) PFU group showed anorexia, lethargy and degression in locomotor activity. This group of mice showed significant decrease in body weight than that of other groups. Colon tissues were collected 14 days after infection. Histological examination revealed that the mucous layer became thinner in the proximal colon and increased number of lymphoid follicles in distal colon in infected animals. The changes in the mucosal structure was most prominent in the group 10(5) PFU MCMV. Viral DNA was present in the colon by in situ hybridization for IE1 gene, and viral gB transcript was positive by RT-PCR. One of the viral major proteins, pp65, was widely distributed in the colon by immunohistochemistry. These data demonstrated that MCMV established infection in colon of the mice after allogenetic skin transplantation. Electron microscopy showed that there were herpes virus particles in the colon tissue.

CONCLUSIONInfection with MCMV in mouse after allogenetic skin transplantation by nasal cavity inoculation resulted in the pathological changes in colon tissue similar to that of inflammation in human colon. The small animal model of colon inflammation may provide a platform for further study of pathogenesis as well as medical intervention of HCMV involved inflammation of human bowel.

Animals ; Colon ; immunology ; pathology ; virology ; Cytomegalovirus Infections ; immunology ; pathology ; virology ; Disease Models, Animal ; Female ; Herpesviridae Infections ; immunology ; pathology ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Muromegalovirus ; genetics ; immunology ; isolation & purification ; Random Allocation ; Skin Transplantation ; adverse effects ; immunology ; pathology ; Transplantation, Homologous ; adverse effects ; immunology ; pathology ; Viral Proteins ; genetics ; metabolism

The human colorectal carcinoma-associated GA733 antigen epithelial cell adhesion molecule (EpCAM) was initially described as a cell surface protein selectively expressed in some myeloid cancers. Gangliosides are sialic acid-containing glycosphingolipids involved in inflammation and oncogenesis. We have demonstrated that treatment with anti-EpCAM mAb and RAW264.7 cells significant inhibited the cell growth in SW620 cancer cells, but neither anti-EpCAM mAb nor RAW264.7 cells alone induced cytotoxicity. The relationship between ganglioside expression and the anti-cancer effects of anti-EpCAM mAb and RAW264.7 was investigated by high-performance thin-layer chromatography. The results demonstrated that expression of GM1 and GD1a significantly increased in the ability of anti-EpCAM to inhibit cell growth in SW620 cells. Anti-EpCAM mAb treatment increased the expression of anti-apoptotic proteins such as Bcl-2, but the expression of pro-apoptotic proteins Bax, TNF-alpha, caspase-3, cleaved caspase-3, and cleaved caspase-8 were unaltered. We observed that anti-EpCAM mAb significantly inhibited the growth of colon tumors, as determined by a decrease in tumor volume and weight. The expression of anti-apoptotic protein was inhibited by treatment with anti-EpCAM mAb, whereas the expression of pro-apoptotic proteins was increased. These results suggest that GD1a and GM1 were closely related to anticancer effects of anti-EpCAM mAb. In light of these results, further clinical investigation should be conducted on anti-EpCAM mAb to determine its possible chemopreventive and/or therapeutic efficacy against human colon cancer.
Animals ; Antibodies, Monoclonal/*immunology/*therapeutic use ; Antigens, Neoplasm/*immunology ; Apoptosis/drug effects ; Cell Adhesion Molecules/*immunology ; Cell Line ; Cell Line, Tumor ; Cell Proliferation/drug effects ; Colon/drug effects/immunology/metabolism/pathology ; Colonic Neoplasms/*drug therapy/genetics/*immunology/pathology ; Gangliosides/genetics/*immunology ; Gene Expression Regulation, Neoplastic/drug effects ; Humans ; Male ; Mice ; Mice, Inbred BALB C

Schwannomas are rare tumors derived from the cells of Schwann which form the neural sheath. Some patients with gastrointestinal schwannoma have been previously reported in the literature. However, schwannomas of the colon are extremely rare. We herein describe a case of schwannoma of the colon. A 49-year-old woman was admitted with complaint of abdominal pain and investigations revealed the presence of a 4 cm sized mass in the ascending colon. Following right hemicolectomy, histopathology and immunohistochemistry confirmed the colonic lesion to be a benign schwannoma. There was no evidence of specific complication or recurrence until now.
Colon, Ascending/*pathology ; Colonic Neoplasms/*diagnosis/pathology/ultrasonography ; Female ; Humans ; Middle Aged ; Neurilemmoma/*diagnosis/pathology/ultrasonography ; S100 Proteins/analysis/immunology ; Tomography, X-Ray Computed

OBJECTIVETo observe the therapeutic effect of Chinese compound, Changlu Enema (CE), on immune ulcerative colitis (UC) in mice.

METHODSMice were randomly divided into the normal group, the model group, the CE high dose (CE-H) and low dose (CE-L) group and the salicylazosulfapyridine (SASP) group. All mice, except those in the normal group, were made into UC model by colonic mucosa protein immunization. After 21 days of medication, the changes of UC activity index and body weight in mice were observed, and the condition of colonic inflammation and histomorphological changes in colonic tissue were observed also.

RESULTSUC activity index was lower and body weight was higher in the two CE groups than those in the model group respectively, showing significant difference (P < 0.05 or P < 0.01); pathological examination showed the pathological changes and inflammatory response in colonic tissue were relieved significantly after treatment, and the improvement in the CE-H group was better than that in the SASP group.

CONCLUSIONTCM compound CE is markedly effective in treating UC rats.

Administration, Rectal ; Animals ; Colitis, Ulcerative ; drug therapy ; immunology ; Colon ; drug effects ; pathology ; Drugs, Chinese Herbal ; administration & dosage ; therapeutic use ; Male ; Mice ; Phytotherapy ; Random Allocation ; Treatment Outcome

OBJECTIVETo investigate the relationship between various Chinese medicine (CM) types and T-cell subsets (CD4(+) and CD8(+)) in the colonic mucous membranes of patients with ulcerative colitis (UC).

METHODSFifty UC patients were enrolled, after differentiation into four types by CM syndromes, i.e., the internal heat-damp accumulation type (IHDA), the qi-stagnancy with blood stasis type (QSBS), the Pi-Shen yang-deficiency type (PSYD) and the yin-blood deficiency type (YBD). From every patient, 3-5 pieces of intestinal mucous membrane tissues were taken through colonoscopy to determine the levels of the T-cell subsets (CD4(+) and CD8(+)) using immunohistochemical method. The results were compared with those in the normal control.

RESULTSThe level of CD8(+)increased and the ratio of CD4(+)/CD8(+)decreased mainly in colonic mucous membranous tissues in UC patients. The level of CD4(+)decreased significantly in IHDA types (P<0.01), but decreased only slightly in the PSYD, QSBS and YBD types. CD8(+)increased significantly in the IHDA types (P<0.01), but only slightly in the other three types.

CONCLUSIONThe IHDA types of UC are closely related with T-cell subsets. The difference of T-cell subsets in various IHDA types of UC patients has provided a theoretical basis for syndrome differentiation in the CM typing of UC.

Adolescent ; Adult ; Aged ; Biopsy ; Blood Circulation ; CD4-CD8 Ratio ; Colitis, Ulcerative ; immunology ; pathology ; Colon ; drug effects ; immunology ; pathology ; Colonoscopy ; Female ; Humans ; Immunohistochemistry ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Qi ; T-Lymphocyte Subsets ; drug effects ; Yang Deficiency ; immunology ; pathology ; Yin Deficiency ; immunology ; pathology ; Young Adult

BACKGROUND/AIMS: This study investigated the expression of T cell immunoglobulin- and mucin-domain-containing molecule 3 (TIM-3), human beta-defensin (HBD)-2, forkhead box protein 3 (FOXP3), and the frequency of CD4+ CD25+ FOXP3+ regulatory T cells (Tregs) in children with Crohn's disease (CD) during infliximab therapy. METHODS: We enrolled 20 CD patients who received infliximab treatment for 1 year. Peripheral blood and colonic mucosal specimens were collected from all CD patients and from healthy control individuals. RESULTS: A significant difference in TIM-3 mRNA expression was evident in peripheral blood mononuclear cells and colonic mucosa between CD patients before infliximab therapy and the healthy controls (p<0.001 and p=0.005, respectively). A significant difference in HBD-2 mRNA expression was found in colonic mucosa between CD patients before infliximab therapy and the healthy controls (p=0.013). In the active phase of CD, at baseline, the median percentage of T cells that were CD25+ FOXP3+ was 1.5% (range, 0.32% to 3.49%), which increased after inflixmab treatment for 1 year to 2.2% (range, 0.54% to 5.02%) (p=0.008). CONCLUSIONS: Our study suggests that both the adaptive and innate immune systems are closely linked to each other in CD pathogenesis. And the results of our study indicate that it could be a useful therapeutic tool, where restoration of TIM-3, HBD-2 and the function of Tregs may repair the dysfunctional immunoregulation in CD.
Adolescent ; Case-Control Studies ; Colon/immunology ; Crohn Disease/*drug therapy/immunology/*metabolism ; Female ; Forkhead Transcription Factors/*metabolism ; Gastrointestinal Agents/*therapeutic use ; Humans ; Infliximab/*therapeutic use ; Intestinal Mucosa/immunology ; Leukocytes, Mononuclear/*metabolism ; Male ; Membrane Proteins/*metabolism ; T-Lymphocytes, Regulatory/immunology ; beta-Defensins/*metabolism