ABSTRACT
Fecal microbiota transplantation (FMT) can induce clinical remission in ulcerative colitis (UC) patients. Enemas, nasoduodenal tubes, and colonoscopies are the most common routes for FMT administration. However, there is a lack of definitive evidence regarding the effectiveness of capsulized FMT treatment in UC patients. In this study, we administered capsulized FMT to 22 patients with active UC to assess the efficiency of capsulized FMT and determine the specific bacteria and metabolite factors associated with the response to clinical remission. Our results showed that the use of capsulized FMT was successful in the treatment of UC patients. Capsulized FMT induced clinical remission and clinical response in 57.1% (12 of 21) and 76.2% (16 of 21) of UC patients, respectively. Gut bacterial richness was increased after FMT in patients who achieved remission. Patients in remission after FMT exhibited enrichment of Alistipes sp. and Odoribacter splanchnicus, along with increased levels of indolelactic acid. Patients who did not achieve remission exhibited enrichment of Escherichia coli and Klebsiella and increased levels of biosynthesis of 12,13-DiHOME (12,13-dihydroxy-9Z-octadecenoic acid) and lipopolysaccharides. Furthermore, we identified a relationship between specific bacteria and metabolites and the induction of remission in patients. These findings may provide new insights into FMT in UC treatment and provide reference information about therapeutic microbial manipulation of FMT to enhance its effects. (This study has been registered at ClinicalTrails.gov under registration no. NCT03426683). IMPORTANCE Fecal microbiota transplantation has been successfully used in patients. Recently, capsulized FMT was reported to induce a response in patients with UC. However, limited patients were enrolled in such studies, and the functional factors of capsulized FMT have not been reported in the remission of patients with UC. In this study, we prospectively recruited patients with UC to receive capsulized FMT. First, we found that capsulized FMT could induce clinical remission in 57.1% of patients and clinical response in 76.2% after 12 weeks, which was more acceptable. Second, we found a relationship between the decrease of opportunistic pathogen and lipopolysaccharide synthesis in patients in remission after capsulized FMT. We also identified an association between specific bacteria and metabolites and remission induction in patients after capsulized FMT. These findings put forward a possibility for patients to receive FMT at home and provide reference information about therapeutic microbial manipulation of FMT to enhance its effects.
Subject(s)
Colitis, Ulcerative , Communicable Diseases , Gastrointestinal Microbiome , Humans , Bacteria , Colitis, Ulcerative/therapy , Colitis, Ulcerative/microbiology , Fecal Microbiota Transplantation/methods , Feces/microbiology , Treatment OutcomeABSTRACT
BACKGROUND AND AIMS: Endoscopy is increasingly performed for evaluating patients with ulcerative colitis (UC). However, its diagnostic accuracy is largely affected by the subjectivity of endoscopists' experience and scoring methods, and scoring of selected endoscopic images cannot reflect the inflammation of the entire intestine. We aimed to develop an automatic scoring system using deep-learning technology for consistent and objective scoring of endoscopic images and full-length endoscopic videos of patients with UC. METHODS: We collected 5875 endoscopic images and 20 full-length videos from 332 patients with UC who underwent colonoscopy between January 2017 and March 2021. We trained the artificial intelligence (AI) scoring system using these images, which was then used for full-length video scoring. To more accurately assess and visualize the full-length intestinal inflammation, we divided the large intestine into a fixed number of "areas" (cecum, 20; transverse colon, 20; descending colon, 20; sigmoid colon, 15; rectum, 10). The scoring system automatically scored inflammatory severity of 85 areas from every video and generated a visualized result of full-length intestinal inflammatory activity. RESULTS: Compared with endoscopist scoring, the trained convolutional neural network achieved 86.54% accuracy in the Mayo-scored task, whereas the kappa coefficient was .813 (95% confidence interval [CI], .782-.844). The metrics of the Ulcerative Colitis Endoscopic Index of Severity-scored task were encouraging, with accuracies of 90.7%, 84.6%, and 77.7% and kappa coefficients of .822 (95% CI, .788-.855), .784 (95% CI, .744-.823), and .702 (95% CI, .612-.793) for vascular pattern, erosions and ulcers, and bleeding, respectively. The AI scoring system predicted each bowel segment's score and displayed distribution of inflammatory activity in the entire large intestine using a 2-dimensional colorized image. CONCLUSIONS: We established a novel deep learning-based scoring system to evaluate endoscopic images from patients with UC, which can also accurately describe the severity and distribution of inflammatory activity through full-length intestinal endoscopic videos.
Subject(s)
Colitis, Ulcerative , Deep Learning , Humans , Colitis, Ulcerative/diagnostic imaging , Artificial Intelligence , Colonoscopy , Inflammation , Computers , Severity of Illness Index , Intestinal MucosaABSTRACT
Background: Grading of endoscopic lesions is important for determining the severity of ulcerative colitis and developing treatment strategies, but the commonly used methods are not sufficient. Objectives: This study aimed to investigate whether new endoscopic scoring systems incorporating lesions and disease extent are associated with clinical disease severity and maintainable remission. Design: This was a retrospective study. In all, 110 patients with ulcerative colitis were included and 87 completed 12-month follow-up. Methods: Colonoscopy was performed within 1 week before blood samples were taken. Degree of ulcerative colitis burden of luminal inflammation (DUBLIN) scores were calculated as the product of Mayo endoscopic score (MES) by disease extent and ulcerative colitis endoscopic index of severity was used to replace MES when calculating modified DUBLIN scores. Results: DUBLIN and modified DUBLIN scores were increased in the moderate and severe groups significantly (p < 0.05). Both of increased scores contributed to the detection of serious diseases, and the clinical cutoff values of DUBLIN and modified DUBLIN were 3[area under the curve (AUC) = 0.809, p = 0.001) and 7(AUC = 0.815, p = 0.001), respectively. They were with high sensitivity, but the specificity of DUBLIN was lower. Both scores were correlated to partial Mayo scores, C-reactive protein and erythrocyte sedimentation rate positively, and they were correlated to the albumin negatively (p < 0.05). Higher modified DUBLIN scores (>7) were associated with an increased risk of treatment failure (hazard ratio = 4.96, 95% confidence interval: 1.17-21.00, p = 0.03), but there were no association between DUBLIN scores and long-term remission (p > 0.05). Conclusion: Increased DUBLIN and modified DUBLIN scores were conducive to screening serious disease, but only modified DUBLIN scores had the potential to assist in making an upgraded therapeutic schedule.
ABSTRACT
This study aimed to investigate whether serum indicators related to iron stores in the body are associated with clinical and endoscopic disease severity. Eighty-four patients with Crohn's disease (CD) and twenty-four healthy volunteers were included. The indicators related to iron stores were detected within one week after endoscopic and CT enterography examinations. Patients were divided into three groups according to the CDAI(Crohn's disease activity index)scores. Serum iron levels were decreased in all groups (p < 0.05), and the values of remission group were higher than those of moderate group (p < 0.001). The total iron binding capacity(TIBC)values of the moderate group were lower than those of the controls and the other groups (p < 0.05). None of the indicators differed significantly among the patients classified by SES-CD (p > 0.05). Underweight, decreased serum iron and TIBC were independent risk factors for moderate clinical disease. Combined detection of decreased serum iron and TIBC was helpful in differentiating severe patients. The sensitivity and specificity were 32.7% and 100%, respectively (AUC = 0.812, p < 0.01). Decreases in serum iron and TIBC were associated with the clinical activity of CD. Combined detection of the two indicators was conducive to screening serious disease.
Subject(s)
Crohn Disease , Crohn Disease/diagnosis , Endoscopy , Humans , Iron , Sensitivity and Specificity , Severity of Illness IndexABSTRACT
BACKGROUND: To identify risk factors for hypovitaminosis D in inflammatory bowel disease and conduct a comprehensive systematic review with meta-analysis to quantify the impact on vitamin D deficiency. METHODS: We conducted a literature search of studies through PubMed, Embase, Cochrane Library, and Web of Science. In addition, relevant articles were searched manually. Studies were included if the odds ratios (OR) and 95% CI of each risk factor were reported or could be calculated. We will use the fixed-effects or random-effects model to estimate the pooled effect. RESULTS: Out of 1018 articles, 25 eligible studies were identified, including 5826 participants. The risk factors associated with hypovitaminosis D were non-Caucasian (OR: 3.79, 95% CI: 2.68-5.34), Crohn's disease (OR: 1.38, 95% CI: 1.21-1.56), disease activity (OR: 1.85, 95% CI: 1.61-2.13), inflammatory bowel disease-related surgery (OR: 1.61, 95% CI: 1.38-1.89), exposure to steroid (OR: 1.61, 95% CI: 1.28-2.03), and biologics (OR: 1.78, 95% CI: 1.48-2.14). In 30 ng/mL and adjusted OR subgroup, male (OR: 1.84, 95% CI: 1.47-2.31) and winter season (OR: 2.49, 95% CI: 1.69-3.67) also were risk factors, respectively. 5-aminosalicylic acid (OR: 1.10, 95% CI: 0.74-1.63) and smoking (OR: 1.19, 95% CI: 0.98-1.45) were unrelated to vitamin D deficiency. CONCLUSIONS: For vitamin D deficiency in inflammatory bowel disease, non-Caucasian, Crohn's disease, disease activity, surgery, exposure to steroid and biologics, males are risk factors, while 5-aminosalicylic acid and smoking are not. The relationship between body mass index, winter season, exposure to immunomodulators, and vitamin D deficiency remains unclear.
Subject(s)
Colitis , Crohn Disease/complications , Inflammatory Bowel Diseases/blood , Vitamin D Deficiency/blood , Vitamin D/blood , Biological Products , Crohn Disease/epidemiology , Humans , Inflammatory Bowel Diseases/complications , Male , Mesalamine , Quality of Life , Risk Factors , Vitamin D Deficiency/epidemiology , Vitamins/bloodABSTRACT
BACKGROUND: Data from single-center experience or small sample-sized studies have shown that chromoendoscopy (CE) might be superior to white-light endoscopy (WLE) for dysplasia surveillance in ulcerative colitis (UC) patients. We performed a prospective randomized trial with a long-term follow-up to compare the detection rate of dysplasia among WLE with targeted biopsies (WLT), WLE with random biopsies (WLR), and dye-based CE with targeted biopsies (CET) in UC patients. METHODS: Patients with long-standing UC were enrolled from 11 medical centers from March 2012 to December 2013 and randomized into three arms (WLT, WLR, and CET). Only high-definition endoscopy was used in all three groups. The patients were followed up by annual endoscopy with biopsies through December 2017. RESULTS: With a median follow-up time of 55 months, a total of 122 patients with 447 colonoscopies were finally analysed in the per-protocol set: WLT (n = 43), WLR (n = 40), and CET (n = 39). A total of 34 dysplastic lesions were found in 29 colonoscopies of 21 patients. WLR and CET could identify more colonoscopies that diagnosed dysplasia than WLT (8.1% and 9.7% vs 1.9%; P = 0.014 and 0.004, respectively). WLR obtained more biopsied samples than WLT and CET (16.4 ± 5.1 vs 4.3 ± 1.4 and 4.3 ± 1.4; both P < 0.001). During the second half of the follow-up (37 - 69 months), CET could identify more colonoscopies that diagnosed dysplasia than WLT (13.3% vs 1.6%, P = 0.015) and showed a trend for increasing the detection rate compared with WLR (13.3% vs 4.9%, P = 0.107). CONCLUSIONS: For a better outcome of cancer/dysplasia surveillance in patients with long-standing UC, CET appeared to be more effective than WLT and less tedious than WLR. CET was found to be particularly useful when a long-term (>3 years) follow-up was conducted for dysplasia surveillance. The trial was registered on www.chictr.org.cn (ChiCTR1900023689).
ABSTRACT
BACKGROUND: This study was to investigate the cytokines and phenotype of macrophages pre-treated with class A1 scavenger receptor (SR-A1) antibody in vitro and the influence on apoptotic pathway of colonic epithelial cells, and to explore the role of SR-A1 mediated macrophages in impaired intestinal barrier of inflammatory bowel diseases (IBDs). METHODS: Mouse macrophage RAW264.7 was pre-treated with SR-A1 antibody in the presence of lipopolysaccharide (LPS). Transwell system was employed for co-culture of RAW264.7 and Caco-2 in the presence of LPS and IFN-γ, with or without SR-A1 antibody pre-treatment. The percentage of F4/80+CD11c+ macrophages, apoptosis rate of Caco-2 cells, and expression of apoptosis and tight junction proteins in Caco-2 cells was determined. RESULTS: Pre-treatment with SR-A1 antibody up-regulated IL-10 expression in RAW264.7, whereas down-regulated the expression of TNF and iNOS. Immunofluorescence staining indicated the upregulation of NF-κB p-p56 after LPS stimulation was significantly inhibited in the presence of SR-A1 antibody. The increase in p-JNK expression was inhibited by SR-A1 antibody. Transwell assay showed the percentage of F4/80+CD11c+ macrophages and apoptotic Caco-2 cells increased after treatment with LPS and IFN-γ, which could be reversed in the presence of SR-A1 antibody. The induction of cleaved caspase-3 and claudin-1 in Caco-2 cells was also suppressed when SR-A1 antibody pre-treatment. CONCLUSIONS: Pre-treatment with SR-A1 antibody can inhibit inflammatory response in LPS-induced macrophages in a NF-κB dependent manner. Pre-treatment with SR-A1 antibody also inhibits M1 phenotype expression of macrophages, and attenuates the pro-apoptotic effect on colonic epithelial cells and disruption of intestinal barrier integrity induced by macrophages.
ABSTRACT
AIM: This study aimed to investigate the anti-inflammatory mechanism of IL-25 mediated mesenchymal stem cells (MSC) treatment for inflammatory bowel disease (IBD) in a DSS-induced rat colitis model. METHODS: Rats with DSS-induced colitis were divided into control and treatment groups: normal control group (rats fed with water), DSS group (rats fed with DSS solution), MSC group (DSS-treated rats injected intravenously with GFP-MSCs), IL-25-MSC group (DSS-treated rats injected intravenously with IL-25 primed GFP-MSCs), and mesalazine group (DSS-treated rats fed with mesalazine). RESULTS: In IL-25-MSC group, therapeutic efficacy (clinical symptoms) was better than in MSC group, but comparable to mesalazine group. In IL-25-MSC group and mesalazine group, fewer infiltrating inflammatory cells and lower pathological score were observed in the intestine. The FOXP3+ cells and IL-4+ cells decreased, but IL-17A+ cells and IFN-γ+ cells increased in the peripheral blood and colonic mucosa after DSS induced colitis, and these phenomena were reversed by MSC or mesalazine treatment. IL-17A+ cells reduced and FOXP3+ cells increased in IL-25-MSC group as compared with MSC group. The expressions of Ki67 and LGR5 were significantly elevated in MSC treatment groups as compared with normal control group, DSS group, and mesalazine group. Definite GFP positive cells were not observed in the intestine of MSC-treated rats. CONCLUSION: IL-25 primed MSCs exert improved therapeutic effects on the intestinal inflammation of IBD rats which may be related to the inhibition of Th17 immune response and induction of T Regulatory cell phenotype. Thus, IL-25 may be an attractive candidate for MSC-based therapy of IBD.
ABSTRACT
BACKGROUND: This study aimed to investigate the influence of IL-25 on the capacity of mesenchymal stem cells (MSCs) to induce intestinal epithelial cell regeneration. METHODS: The CD4+IL-25R+ cells and LGR5+IL-25R+ cells in colonic mucosa of Crohn's disease (CD) patients, ulcerative colitis (UC) patients and healthy controls were detected by immunofluorescence staining, and the CD4+IL-25R+ cells in peripheral blood were detected by flow cytometry. Rat MSCs were separated and stimulated with IL-25. Then, MSCs were further incubated in IL-25-free DMEM for 24 h, and this DMEM was collected as conditioned medium (CM). IEC-6 cells were divided into 3 groups: experimental group (CM and TNF-α), control group (DMEM and TNF-α) and negative control group (DMEM). RESULTS: The CD4+IL-25R+ cells and LGR5+IL-25R+ cells significantly increased in the colonic mucosa of active CD patients and UC patients compared with IBD patients in remission and healthy controls. The CD4+IL-25R+ cells reduced in peripheral blood of IBD patients, which was inversely correlated with inflammatory markers (ESR and CRP). CM facilitated the migration and proliferation of IEC-6 cells in the presence of TNF-α. The protein expression of AKT, p38 and ERK increased in IEC-6 cells after treatment with CM and TNF-α. CONCLUSION: IL-25R is involved in Th-related mucosal inflammation and proliferation of intestinal stem cells in IBD. IL-25 enhances the capacity of MSC to induce intestinal epithelial cell regeneration, and MSC therapy with IL-25 may be a new direction for IBD treatment.
ABSTRACT
The trefoil factor family (TFF) is a group of short secretory peptides of gastric mucous neck cells. The loss of TFF2 protein expression enhances gastric inflammation and occurs in gastric cancer. In this study, we examined the effect of TFF2 on gastric cancer cell lines in vitro and characterized the interaction between TFF2 and Sp3, including the mechanisms that mediate this interaction, using genomics and proteomics approaches, as well as genetics techniques, such as RNA interference and gene knockdown. Assays were performed to examine the role of TFF2 and Sp3 in cancer cell proliferation, invasion and migration. We found that TFF2 expression inhibited the proliferation and invasion capacity of gastric cancer cells, and induced apoptosis. TFF2 interacted with the Sp3 protein, as shown by immunoï¬uorescence staining and immunoprecipitation with western blot analysis. Sp3 knockdown in gastric cancer cells antagonized TFF2 antitumor activity. Additionally, TFF2 upregulated the expression of pro-apoptotic proteins, such as Bid, but downregulated the expression of NF-κB and the anti-apoptotic proteins, Bcl-xL and Mcl1. By contrast, Sp3 knockdown significantly blocked TFF2 activity, affecting the expression of these proteins. The data from our study demonstrate that the antitumor activity of TFF2 is mediated by an interaction with the Sp3 protein in gastric cancer cells. Additional in vivo and ex vivo warrned in order to fully characterize this interaction.
Subject(s)
Cell Proliferation/genetics , Gene Expression Regulation, Neoplastic , Sp3 Transcription Factor/genetics , Trefoil Factor-2/genetics , Apoptosis/genetics , BH3 Interacting Domain Death Agonist Protein/metabolism , Blotting, Western , Cell Line , Cell Line, Tumor , Cell Movement/genetics , HEK293 Cells , Humans , Microscopy, Video , Myeloid Cell Leukemia Sequence 1 Protein/metabolism , NF-kappa B/metabolism , Neoplasm Invasiveness , Protein Binding , RNA Interference , Sp3 Transcription Factor/metabolism , Stomach Neoplasms/genetics , Stomach Neoplasms/metabolism , Stomach Neoplasms/pathology , Trefoil Factor-2/metabolism , bcl-X Protein/metabolismABSTRACT
Trefoil factor family 2 (TFF2) participates in mucus stabilization and repair, apoptosis, and inflammatory responses. Previously published reports have indicated that several growth factors and basal transcription factors are associated with the expression of TFF2. However, the detailed mechanisms that regulate TFF2 expression are not fully understood. The present study was designed to assess the essential role of the transcription factor SP3 with respect to TFF2 expression. We first demonstrated that there was a negative correlation between the expression levels of SP3 and TFF2. Thus, in the examined cells, the overexpression of SP3 decreased the expression level of TFF2, whereas the inhibition of SP3 increased the expression level of TFF2. Moreover, we discovered two GC boxes in the TFF2 promoter and confirmed the specific binding of SP3 to this promoter. On the whole, this study indicated that Sp3 was a major regulator of TFF2 expression. This knowledge should contribute to our understanding of the role that is played by SP3 in the regulation of TFF2 expression.
Subject(s)
Gene Expression Regulation , Peptides/genetics , Sp3 Transcription Factor/metabolism , Cell Line, Tumor , Humans , Peptides/metabolism , Promoter Regions, Genetic , Sp3 Transcription Factor/genetics , Transcription, Genetic , Transcriptional Activation , Trefoil Factor-2ABSTRACT
BACKGROUND/AIMS: The aim of this study was to investigate the diagnostic yield of double-balloon enteroscopy (DBE) and its impact on subsequent management. METHODOLOGY: This study is a retrospective analysis of 300 consecutive patients for investigation of small bowel disease that had been suspected by both clinical symptoms and imaging tests. The final management was guided by the results of DBE. Demographic, clinical, procedural and outcome data were collected for analysis. RESULTS: Among the 300 patients, lesions were found in 213 (70.1%). These lesions were small-intestinal ulcers, Crohn's disease, chronic inflammation, Meckel's diverticulae, angiodysplasia, polyps, ulcerative lipoma and tumor. Ninety-nine of the 115 patients with suspected intestinal hemorrhage were confirmed, with a positive rate of 86.1%. Also confirmed were 60 of the 99 patients with abdominal pain (positive rate of 60.6%); sixteen of 22 patients with chronic diarrhea (positive rate of 72.7%); thirty-three of the 55 patients with abdominal distention or malnutrition (positive rate of 60.0%); and 7 of 9 patients with refractory hypoalbuminemia. Treatment was performed in 2.3% of patients (3 polyps and 2 angiodysplasia). CONCLUSIONS: This large pilot series shows that DBE is a safe and feasible diagnostic and therapeutic tool for suspected or documented small-bowel diseases.
Subject(s)
Double-Balloon Enteroscopy/methods , Intestinal Diseases/diagnosis , Intestine, Small/pathology , Adolescent , Adult , Aged , Female , Humans , Intestinal Diseases/therapy , Male , Middle AgedABSTRACT
BACKGROUND AND AIM: Hepatitis B virus (HBV) infection poses great challenges to humans, claiming one million lives annually worldwide. Solid data have related HBV to hepatocellular carcinoma. METHODS: In the present research, we verified the interaction between surface protein (HBs) encoded by HBV and aldolase A (ALDA) using yeast two-hybrid, mammalian two-hybrid, co-immunoprecipitation, GST pull-down and laser scanning confocal. RESULTS: Anti-ALDA antibody precipitated Gal4-HBs fusion protein in the presence of HBs. Anti-HBs antibody precipitated p65ΔN-ALDA only in the presence of ALDA. Small HBs could be pulled down by GST-ALDA. Cells transfected with pCMV-AD-ALDA showed a protection from ultraviolet radiation-induced apoptosis (21.3% ± 1.3% for ALDA, 35.4% ± 2.1% for control, P < 0.05). CONCLUSIONS: An interaction does exist between ALDA and HBs. The S region within HBs is sufficient for binding ALDA. In addition, ALDA conferred protection to ultraviolet radiation-induced apoptosis, and this effect was enhanced by the interaction between HBs and ALDA.
Subject(s)
Apoptosis/radiation effects , Carcinoma, Hepatocellular/immunology , Fructose-Bisphosphate Aldolase/immunology , Hepatitis B Surface Antigens/immunology , Hepatitis B, Chronic/immunology , Liver Neoplasms/immunology , Ultraviolet Rays , Animals , Apoptosis/immunology , Blotting, Western , Carcinoma, Hepatocellular/genetics , Carcinoma, Hepatocellular/pathology , Cell Line , Cricetinae , DNA/analysis , DNA, Neoplasm/genetics , Flow Cytometry , Hepatitis B, Chronic/genetics , Hepatitis B, Chronic/pathology , Humans , Liver Neoplasms/genetics , Liver Neoplasms/pathology , Microscopy, Confocal , Polymerase Chain ReactionABSTRACT
AIMS: To study the expression of human intestinal trefoil factor (hITF) mRNA in Crohn's disease and to screen the cellular proteins that can interact with the hITF protein by a yeast two-hybrid system in order to explore the mechanism of hITF in protecting intestinal mucosa from injury. METHODS: Seventy-eight patients underwent double-balloon enteroscopy (DBE). Expression of hITF mRNA was detected by quantitative real-time polymerase chain reaction analysis (qRT-PCR). The hITF gene was amplified by PCR and cloned into vector pDEST32. The yeast cells cotransformed with pDEST32-hITF and the human jejunal cDNA library were plated in a selective SC-Leu-Trp-His-Ura medium. The subsequent screen was performed with χ-gal detection, and true-positive clones were sequenced and analyzed with bioinformatics. Co-immunoprecipitation (Co-IP) was performed to confirm the binding of putative proteins to the hITF protein. RESULTS: Thirty-nine patients were diagnosed with Crohn's disease. We found that the expression of hITF mRNA is significantly increased in Crohn's disease compared to normal controls. A total of ten colonies were selected and sequenced. Among these, six colonies were Homo sapiens zinc finger protein 193 (ZNF193), three colonies were Homo sapiens Aldo-keto reductase family 1C 1 (AKR1C1), and one colony was of an unknown gene. A reverse two-hybrid experiment and Co-IP indicated that ZNF193 and AKR1C1 might interact with hITF. CONCLUSIONS: The expression of hITF mRNA is increased in Crohn's disease. ZNF193 and AKR1C1 are proteins that can interact with the hITF protein by a yeast two-hybrid system and Co-IP, hITF may contribute to the mucosal repair through this interaction.
Subject(s)
Crohn Disease/genetics , Genetic Testing/methods , Peptides/genetics , Plasmids/genetics , Two-Hybrid System Techniques , Cell Line , Crohn Disease/metabolism , Crohn Disease/pathology , Gene Expression/physiology , Gene Library , Homeostasis/physiology , Humans , Intestinal Mucosa/pathology , Intestinal Mucosa/physiology , Jejunum/pathology , Jejunum/physiology , Kidney/cytology , RNA, Messenger/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Trefoil Factor-2 , Up-Regulation/physiologyABSTRACT
Jumping translocation breakpoint protein (JTB) is suppressed in many cancers, implying it plays a role in the neoplastic transformation of cells. In order to explore the role of JTB in the carcinogenesis of liver, we used mammalian two-hybrid, co-immunoprecipitation, GST pull-down and laser scanning confocal to verify the interaction between HBs and JTB. According to the results, HBs interacts with JTB. In addition, we further determined that S region within HBs is sufficient for binding JTB. Overexpression of JTB conferred resistance to apoptosis induced by ultraviolet radiation, whereas this effect was compromised by the co-overexpression of HBs.
Subject(s)
Cell Transformation, Neoplastic/metabolism , Hepatitis B Surface Antigens/metabolism , Liver Neoplasms/metabolism , Membrane Proteins/metabolism , Neoplasm Proteins/metabolism , Radiation Tolerance/physiology , Apoptosis/physiology , Apoptosis/radiation effects , Blotting, Western , Cell Transformation, Neoplastic/genetics , Cell Transformation, Neoplastic/radiation effects , Flow Cytometry , Fluorescent Antibody Technique , Hepatitis B, Chronic/complications , Humans , Immunoprecipitation , Liver Neoplasms/genetics , Liver Neoplasms/virology , Microscopy, Confocal , Polymerase Chain Reaction , Transfection , Two-Hybrid System Techniques , Ultraviolet RaysABSTRACT
BACKGROUND: Transfection in mammalian cells based on liposome presents great challenge for biological professionals. To protect themselves from exogenous insults, mammalian cells tend to manifest poor transfection efficiency. In order to gain high efficiency, we have to optimize several conditions of transfection, such as amount of liposome, amount of plasmid, and cell density at transfection. However, this process may be time-consuming and energy-consuming. Fortunately, several mathematical methods, developed in the past decades, may facilitate the resolution of this issue. This study investigates the possibility of optimizing transfection efficiency by using a method referred to as least-squares support vector machine, which requires only a few experiments and maintains fairly high accuracy. RESULTS: A protocol consists of 15 experiments was performed according to the principle of uniform design. In this protocol, amount of liposome, amount of plasmid, and the number of seeded cells 24 h before transfection were set as independent variables and transfection efficiency was set as dependent variable. A model was deduced from independent variables and their respective dependent variable. Another protocol made up by 10 experiments was performed to test the accuracy of the model. The model manifested a high accuracy. Compared to traditional method, the integrated application of uniform design and least-squares support vector machine greatly reduced the number of required experiments. What's more, higher transfection efficiency was achieved. CONCLUSION: The integrated application of uniform design and least-squares support vector machine is a simple technique for obtaining high transfection efficiency. Using this novel method, the number of required experiments would be greatly cut down while higher efficiency would be gained. Least-squares support vector machine may be applicable to many other problems that need to be optimized.
Subject(s)
Liposomes , Software , Transfection/methods , Algorithms , Cell Line, Transformed , Genetic Vectors , Humans , Least-Squares Analysis , Models, BiologicalABSTRACT
AIM: To study the role of mitochondrial energy disorder in the pathogenesis of ethanol-induced gastric mucosa injury. METHODS: Wistar rats were used in this study. A gastric mucosal injury model was established by giving the rats alcohol. Gross and microscopic appearance of gastric mucosa and ultrastructure of mitochondria were evaluated. Malondiadehyde (MDA) in gastric mucosa was measured with thiobarbituric acid. Expression of ATP synthase (ATPase) subunits 6 and 8 in mitochondrial DNA (mtDNA) was determined by reverse transcription polymerase chain reaction (RT-PCR). RESULTS: The gastric mucosal lesion index was correlated with the MDA content in gastric mucosa. As the concentration of ethanol was elevated and the exposure time to ethanol was extended, the content of MDA in gastric mucosa increased and the extent of damage aggravated. The ultrastructure of mitochondria was positively related to the ethanol concentration and exposure time. The expression of mtDNA ATPase subunits 6 and 8 mRNA declined with the increasing MDA content in gastric mucosa after gavage with ethanol. CONCLUSION: Ethanol-induced gastric mucosa injury is related to oxidative stress, which disturbs energy metabolism of mitochondria and plays a critical role in the pathogenesis of ethanol-induced gastric mucosa injury.
Subject(s)
Energy Metabolism/drug effects , Ethanol/toxicity , Gastric Mucosa/drug effects , Mitochondria/drug effects , Oxidative Stress/drug effects , Animals , DNA, Mitochondrial/metabolism , Dose-Response Relationship, Drug , Gastric Mucosa/metabolism , Gastric Mucosa/ultrastructure , Male , Malondialdehyde/metabolism , Mitochondria/metabolism , Mitochondria/ultrastructure , Mitochondrial Proton-Translocating ATPases/metabolism , Models, Animal , RNA, Messenger/metabolism , Rats , Rats, Wistar , Severity of Illness Index , Thiobarbituric Acid Reactive Substances/metabolism , Time FactorsABSTRACT
AIM: To study the molecular forms of trefoil factor 1 (TFF1) in normal gastric mucosa and its expression in normal and abnormal gastric tissues (gastric carcinoma, atypical hyperplasia and intestinalized gastric mucosa) and the role of TFF1 in the carcinogenesis and progression of gastric carcinoma and its molecular biological mechanism underlying gastric mucosa protection. METHODS: The molecular forms of TFF1 in normal gastric mucosa were observed by Western blot. The expression of TFF1 in normal and abnormal gastric tissues (gastric carcinoma, atypical hyperplasia and intestinalized gastric mucosa) was also assayed by immunohistochemical method. The average positive AO was estimated by Motic Images Advanced 3.0 software. RESULTS: Three patterns of TFF1 were found in normal gastric mucosa: monomer, dimmer, and TFF1 compound whose molecular weight is about 21 kDa. The concentration of TFF1 compound was the highest among these three patterns. TFF1 was expressed mainly in epithelial cytoplasm of the mucosa in gastric body and antrum, especially around the nuclei. The closer the TFF1 to the lumen, the higher the expression of TFF1. The expression of TFF1 in peripheral tissue of gastric carcinoma (0.51 +/- 0.07) was higher than that in normal gastric mucosa (0.44 +/- 0.06, P < 0.001). The expression of TFF1 in gastric adenocarcinoma was positively related to the differentiation of adenocarcinoma. The lower the differentiation of adenocarcinoma was, the weaker the expression of TFF1. No TFF1 was expressed in poorly-differentiated adenocarcinoma. The expression of TFF1 in moderately-well differentiated adenocarcinoma (0.45 +/- 0.07) was a little lower than that in normal mucosa (P > 0.05). The expression of TFF1 in gastric mucosa with atypical hyperplasia (0.57 +/- 0.03) was significantly higher than that in normal gastric mucosa (P < 0.001). No TFF1 was expressed in intestinalized gastric mucosa. There was no statistically significant difference between the expressions of TFF1 in gastric mucosa around the intestinalized tissue (0.45 +/- 0.07) and normal gastric mucosa (P > 0.05). CONCLUSION: TFF1 is expressed mainly in epithelial cytoplasm of the mucosa in gastric body and antrum. Its main pattern is TFF1 compound, which may have a greater biological activity than monomer and dimer. The expression of TFF1 in peripheral mucosa of gastric ulcer is higher than that in mucosa 5 cm beyond the ulcer, indicating that TFF1 plays an important part in protection and restitution of gastric mucosa. The expression of TFF1 is increased in peripheral tissues of gastric carcinoma and gastric mucosa with atypical hyperplasia, but is decreased in cancer tissues, implying that TFF1 may be related to suppression and differentiation of carcinoma. The weaker expression of TFF1 in poorly-differentiated carcinoma may be related to the destruction of glands and cells in cancer tissues and the decrease in secretion of TFF1.
Subject(s)
Gastric Mucosa/metabolism , Stomach Neoplasms/genetics , Tumor Suppressor Proteins/genetics , Adult , Aged , Biopsy , Breast Neoplasms/genetics , Carcinoma/genetics , Carcinoma/pathology , Cell Line, Tumor , Duodenum/cytology , Duodenum/pathology , Female , Gastric Mucosa/abnormalities , Gastric Mucosa/cytology , Gastric Mucosa/pathology , Gene Expression Regulation, Neoplastic , Humans , Hyperplasia/genetics , Middle Aged , Reference Values , Stomach Neoplasms/pathology , Stomach Neoplasms/surgery , Trefoil Factor-1ABSTRACT
OBJECTIVE: To study the molecular forms of TFF1 in normal gastric mucosa and its expression in normal, gastric carcinoma, atypical hyperplasia, and intestinalized gastric mucosa. METHODS: The molecular forms of TFF1 in normal gastric mucosa was observed by western blotting. The expression of TFF1 in normal, gastric carcinoma, atypical hyperplasia, and intestinalization gastric mucosa was assayed immunohistochemically. RESULTS: TFF1 existed in normal gastric mucosa in forms of monomer, dimer and 21-kD TFF1 complex, with the last being the richest. TFF1 was expressed mainly in the epithelial cytoplasm of the mucosa in the gastric body and antrum, especially around the nucleus, and the closer to the lumen, the higher the expression. TFF1 expression in the tissues adjacent to gastric carcinoma was higher than that in normal gastric mucosa (P<0.001), and the expression in gastric adenocarcinoma was positively correlated to differentiation of adenocarcinoma. No TFF1 was expressed in poorly differentiated adenocarcinoma. The expression of TFF1 in moderate and well differentiated adenocarcinoma was a little lower than that in normal mucosa (P>0.05). The gastric mucosa with atypical hyperplasia had significantly higher TFF1 expression than normal gastric mucosa (P<0.001), and TFF1 was not detected in intestinalized gastric mucosa. There was no significant difference in TFF1 expression between gastric mucosa around the intestinalized tissues and normal gastric mucosa (P>0.05). CONCLUSIONS: TFF1 plays an important part in protection and restitution of the gastric mucosa, and TFF1 may be related to suppression and differentiation of carcinoma.
Subject(s)
Gastric Mucosa/metabolism , Stomach Neoplasms/metabolism , Tumor Suppressor Proteins/metabolism , Adenocarcinoma/metabolism , Adenocarcinoma/pathology , Adult , Aged , Blotting, Western , Esophageal Neoplasms/metabolism , Esophageal Neoplasms/pathology , Female , Gastric Mucosa/pathology , Humans , Immunohistochemistry , Male , Middle Aged , Stomach Neoplasms/pathology , Trefoil Factor-1ABSTRACT
OBJECTIVE: To investigate the role of trefoil factor 1 (TFF1) expression in gastric mucosa healing and carcinoma suppression. METHODS: TEF1 expressions in normal and pathological gastric mucosa tissues were detected by immunohistochemical analysis, and the average optical density (OD) was estimated by Motic Images Advanced 3.0 software. RESULTS: Increased TFF1 expression was detected in gastritis, gastric ulcer and duodenal ulcer tissues as compared with that in normal gastric mucosa. TFF1 expression was increased in multiple and compound ulcer in comparison with simple ulcer, but there was no significant difference between gastric ulcer and duodenal ulcer, or between gastritis and simple ulcer tissues. Increased TFF1 was also detected in the mucosa adjacent to the gastric adenocarcinoma, and adenocarcinoma with poorer differentiation had lower TFF1 expression. CONCLUSIONS: TFF1 expression is increased in gastritis, gastric ulcer and duodenal ulcer, and multiple and compound ulcer has higher TFF1 expression than simple ulcer, suggesting the protective role of TFF1 in gastric mucosa and epithelial restitution. TFF1 may directly contribute to the differentiation of adenocarcinoma, and the poorer the differentiation, the lower the expression of TFF1.