Active Ulcerative Colitis Group Research Articles

Possible role of intestinal fungal dysbiosis in dectin-1 and cytokines expression in patients with ulcerative colitis.

Dysregulation of cytokines and intestinal mycobiome has been surveyed in the progression of inflammatory bowel diseases (IBDs), including ulcerative colitis (UC) and Crohn's disease (CD). On the other hand, the intestinal fungal flora and its main receptor, Dectin-1, induce immune-derived cytokines. Total 64 individuals comprising 32 patients with UC (case group) and 32 healthy subjects (HS group) were assessed. The type and prevalence of fecal yeast species were determined by deoxyribonucleic acid (DNA) sequencing through polymerase chain reaction (PCR) amplification usingITS4 and ITS5 primers. Furthermore, the ribonucleic acid (RNAs) of IL-4, IL-10, IL-17, IL-22 and IFN-γ were extracted. The expression of Dectin-1 gene was then measured in the excised tissue samples. A higher global fungal load in UC-affectedpatients (75%) was found in comparison with the HS group (25%), especially Candida albicans. Saccharomyces cerevisiae was significantly reduced in the fecal samples of UC-affected patients compared to HS (15.04% vs. 1.93% UC). The expression level of Dectin-1 was significantly elevated in patients with active UC (7.37 ± 0.81) than in patients with non-active UC (5.01 ± 77.25) and healthy controls (0.97 ± 0.24) (p < 0.05). The expression levels of IL-4, IL-10, especially both IL-17 and IL-22, were higher in the active UC group compared to the HS group (p = 0.0101, p = 0.0155, p < 0.0001, p < 0.0001, respectively). Similar expression level of IL-4, IL-10, IL-17, IL-22 (p > 0.999) and lower expression of interferongamma (IFN-γ) (p = 0.0021) were found in the non-active UC group compared to the HS group. A significant weak to moderate correlation was detected between Dectin-1 and IL-17 (r = 0.339, p = 0.019), as well as Dectin-1 and IL-22 (r = 0.373, p = 0.015). Furthermore, the expression levels of Dectin-1, IL-17 and IL-22 displayed significant associations with disease activity (p < 0.001, p = 0.029 and p = 0.003, respectively), regardless of the participant group. The current study revealed a possible role for intestinal fungi to promote colonic inflammation and increase UC activity through Dectin-1 stimulation. A positive correlation was detected between intestinal fungal richness with UC susceptibility and activity. IL-4 and IL-10 were associated with disease activity. Besides, the expression levels of Dectin-1, IL-17 and IL-22 were independently associated with disease activity.

Monocyte/High-Density Lipoprotein Ratio Is an Indicator of Activity in Patients with Ulcerative Colitis.

In patients with ulcerative colitis, endoscopic and clinical indices are used to assess the disease activity. In addition, stud- ies have been carried out for easier and cheaper markers in recent years. For this purpose, we evaluated the monocyte/high-density lipoprotein ratio of the disease activity. According to clinical activity and partial Mayo scores, a total of 114 patients, 53 in the active ulcerative colitis group and 61 in the ulcerative colitis remission group were included in the study. Monocyte/high-density lipoprotein ratio, neutrophil/lymphocyte ratio, platelet/lymphocyte ratio, C-reactive protein, and erythrocyte sedimentation rate measurements of these 2 groups were recorded. Ulcerative colitis remission group and active ulcerative colitis group were compared in terms of activity. The monocyte/high-density lipoprotein ratio value in the active ulcerative colitis group was significantly higher than that of the ulcerative colitis remission group (10.68 ± 3.39, 6.68 ± 1.39, P < .001, respectively). The monocyte/high-density lipoprotein ratio value for active ulcerative colitis at a cut-off value of 7.4 had 83% sensitivity and 81% specificity. In the active ulcerative colitis group, neutrophil/lymphocyte ratio, C-reactive protein, and erythrocyte sedimentation rate values were significantly higher than the ulcerative colitis remission group (P < .001, P < .001, P < .001, respectively). Monocyte/high-density lipoprotein ratio is an inexpensive and effective marker that can be used to determine the activity of ulcerative colitis.

Inflammatory Indexes for Assessing the Severity and Disease Progression of Ulcerative Colitis: A Single-Center Retrospective Study.

BackgroundActive and severe ulcerative colitis (UC) and non-response to 5-aminosalicylic acid (5-ASA) are related to poor outcomes and should be accurately identified. Several integrated inflammatory indexes are potentially useful to assess the disease severity in patients with acute or critical diseases but are underexplored in patients with UC.MethodsPatients with UC consecutively admitted to our hospital between January 2015 and December 2020 were retrospectively grouped according to the activity and severity of UC and response to 5-ASA. The neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), systemic immune-inflammation index (SII), neutrophil-to-platelet ratio (NPR), platelet-to-albumin ratio (PAR), C-reactive protein-to-albumin ratio (CAR), and C-reactive protein-to-lymphocyte ratio (CLR) were calculated. The areas under receiver operating characteristic curves (AUC) were calculated.ResultsOverall, 187 patients with UC were included, of whom 151 were active, 55 were severe, and 14 were unresponsive to 5-ASA. The active UC group had significantly higher NLR, PLR, SII, and PAR levels. SII had the greatest predictive accuracy for active UC, followed by PLR, PAR, and NLR (AUC = 0.647, 0.641, 0.634, and 0.626). The severe UC group had significantly higher NLR, PLR, SII, PAR, CAR, and CLR levels. CLR had the greatest predictive accuracy for severe UC, followed by CAR, PLR, SII, NLR, and PAR (AUC = 0.732, 0.714, 0.693, 0.669, 0.646, and 0.63). The non-response to the 5-ASA group had significantly higher CAR and CLR levels. CAR had a greater predictive accuracy for non-response to 5-ASA than CLR (AUC = 0.781 and 0.759).ConclusionSII, CLR, and CAR may be useful for assessing the severity and progression of UC, but remain not optimal.

Soluble syndecan-1 as marker of intestinal inflammation: A preliminary study and evaluation of a new panel of biomarkers for non-invasive prediction of active ulcerative colitis.

Syndecan-1 (Sdc1) is a heparin sulfate proteoglycan expressed in intestinal epithelium, which plays a crucial role in inflammation and epithelial repair. Sdc1-knockout mice have a deteriorated course of dextran sulfate sodium-induced colitis as compared to controls. Syndecan-1 is also shed into the serum during inflammation of the epithelium. We hypothesized that an increased serum level of soluble Sdc1 is a biomarker of intestinal inflammation in ulcerative colitis (UC). To evaluate serum soluble Sdc1 as a biomarker of intestinal inflammation in UC. This is a proof-of-concept study. Patients were recruited by the University Hospital Münster and HELIOS Albert Schweitzer Klinik Northeim (Germany). Blood samples were collected from UC patients actively suffering from this condition and those in remission. The levels of Sdc1 were measured with Diaclone CD 138 ELISA kit (Diaclone Research, Besançon, France) and routine clinical data were collected (C-reactive protein (CRP) levels, calprotectin in stool samples). Data were analyzed using SPSS software. Soluble Sdc1 levels were significantly elevated in the active UC group as compared to the inactive UC group (94.5 ±68.1 ng/mL compared to 28.3 ±12.6 ng/mL, p = 0.0020). The levels of Sdc1 also significantly correlated with the severity of UC as measured with the Mayo score (p = 0.0248). Receiver operating characteristic (ROC) analysis showed a good correlation of Sdc1 with an endoscopic Mayo score ≥2, with a value of 0.7747 (95% confidence interval (95% CI) = 0.5775-0.9718). A cutoff value of 37.1 ng/mL of Sdc1 showed a sensitivity of 78% and a specificity of 77%. A panel of biomarkers including CRP, hemoglobin, hematocrit, and Sdc1 was able to precisely predict active UC with an area under the curve (AUC) = 0.9395 (95% CI = 0.8509-1.0000). Serum soluble Sdc1 correlates significantly with mucosa inflammation and Mayo score in UC. Clinical trials No. NCT02333526.

Expression of TOB/BTG family members in patients with inflammatory bowel disease.

In recent years, the role of anti-proliferative TOB proteins in the regulation of immune response by inhibiting T cell activation has been demonstrated. Nevertheless, no previous studies have explored their expression in patients with IBD. The aim of the study was to characterize the gene and protein expression of the TOB/BTG family in intestinal tissue of patients with IBD. This is an observational and cross-sectional study that included 63 IBD patients. Gene expression of TOB/BTG family was measured by RT-PCR. Protein expression of TOB/CD16 and BTG/Ki-67 was evaluated by immunohistochemistry. TOB/BTG family mRNAs were detected and quantitated by RT-qPCR in rectal and ileum biopsies from UC patients and CD patients, respectively, and non-inflammatory control tissues. Results showed that TOB1 and BTG1 gene expression was decreased in the colonic mucosa from patients with UC compared with the control group. The TOB2 and BTG2 genes were over-expressed in the colonic mucosa of patients with UC in remission compared with the active UC and control group. The high TOB2 gene expression was associated with histological remission (P=.01). TOB1/CD16, TOB2/CD16, BTG1/Ki-67, BTG2/Ki-67 and BTG4/Ki-67 single and double positive cells were mostly NK, macrophages, epithelial cells, connective tissue cells and perivascular inflammatory infiltrates in tissues from patients with UC and CD. This is the first depiction of the TOB/BTG family gene and protein expression in rectal and ileum tissues by a CD16+ subpopulation in IBD.

Ülseratif kolitte hastalığın şiddetini öngörmede non-invaziv belirteçler olarak hematolojik parametrelerin değerlendirilmesi

Amaç: Ülseratif kolitte (ÜK) hastalık aktivitesinin değerlendirilmesi ve tedavinin şekillendirilmesi için birçok klinik aktivite indeksi ve non invaziv belirteç araştırılmış fakat hiçbiri inflamatuvar aktivitenin saptanmasında histopatolojik ve endoskopik incelemeler kadar kesin bulgu vermemiştir. Basit ve kolayca elde edilebilen nötrofillerin lenfosit ve plateletlere (N/LP) oranının ÜK’deki klinik hastalık aktivitesi ve hastalığın endoskopik şiddeti arasındaki ilişkiyi incelemeyi amaçladık.Gereç ve Yöntem: Baron skoru ve Klinik Kolit Aktivite İndeksi (CCAİ) bulgularına göre 32 si aktif, 37 si remisyonda toplam 69 ÜK hastası ve 36 sağlıklı katılımcı çalışmaya dahil edildi. Mutlak nötrofil sayısı, NLR, N/LP, MPV, C-reaktif protein ve Eritrosit sedimantasyon hızı ölçüldü ve kaydedildi.Sonuçlar: Aktif ÜK grubunda ve aktif olmayan ÜK grubunda kontrol grubuna kıyasla önemli derecede nötrofil/lenfosit oranında (NLR) yükselme gözlendi (sırasıyla 2,98±1,69, 2,56±1,2, 1,59±0,43; p&amp;lt;0,0001). Fakat aktif ÜK grubu ile remisyon grubu arasında fark olmasına rağmen istatistiksel olarak anlamlı değildi (p&amp;lt;0,284). Aktif ÜK grubunda ve aktif olmayan ÜK grubunda kontrol grubuna kıyasla önemli derecede N/LP yükselmesi gözlendi (sırasıyla 0,96±0,4, 0,9±0,4, 0,7±0,2; p&amp;lt;00001). Fakat aktif ÜK grubu ile remisyon grubu arasında fark olmasına rağmen istatistiksel olarak anlımlı değildi (p=0,588). Sedim ve CRP’nin hastalık aktivitesini ayırt etmek için anlamlı bağımsız belirteçler olduğu bulundu (sırasıyla p&amp;lt;0,05, p&amp;lt;0,05).Tartışma: NLR’ler ve N/LP’ler, kontrol grubuna kıyasla ülseratif kolit hastalarında anlamlı farklılık göstermiştir. Fakat bu parametreler ülseratif kolit hastalarının aktivasyon-remisyon farkını saptamada bir biyobelirteçleri olarak kullanılamayacağı değerlendirilmiştir.

473 The Prevalence and Burden of Rome IV Functional Colorectal Disorders in Ulcerative Colitis

INTRODUCTION: Despite advances in Ulcerative Colitis (UC) therapies, many patients suffer refractory symptoms in the absence of active inflammation. For this group, treatment remains challenging, with a paucity of therapeutic options. In this prospective, ongoing study, we aim to determine the prevalence and burden of functional colorectal disorders inpatients with quiescent UC. METHODS: In a cross-sectional study, consecutive patients with UC attending Inflammatory Bowel Disease (IBD) clinicscompleted a series of validated questionnaires; including Hospital Anxiety and Depression Scale (HADS), the Rome IV diagnostic questionnaire for functional gastrointestinal disorders (FGIDs), an IBD-QoL score and the IBD-control questionnaire. Participants were asked to return a Fecal Calprotectin (FCP) within 2 weeks of completing questionnaires. Quiescent UC was defined as IBD-control 8 score ≥13 and IBD-control-VAS ≥85, and/or FCP levels ≤250 (where available, FCP was used in preference to IBD-control to classify UC activity). Based on Rome IV diagnosis and UC disease activity (active or quiescent), patients were divided into groups and data compared using non-parametric tests. RESULTS: Overall, n = 97 UC patients (n = 50 males, mean age 48 (range 18-82)) participated. 41/97, (42%) UC patients met the Rome IV diagnostic criteria for ≥1 FGIDs (irritable bowel syndrome n = 26, functional constipation n=6 and faecal incontinence (FI) n = 22). Disease activity data (IBD-control and/or FCP) were available for all patients, and based on these 61/97, 63% had quiescent UC. Within the quiescent UC group, 25/61 (41%) met the Rome IV diagnostic criteria for ≥1 FGIDs (irritable bowel syndrome n = 14, functional constipation n = 3 and FI n = 13). In the active UC group, those with co-existing FGIDs, compared to those without FGIDs, had significantly worse median QoL scores (P = 0.02), higher HADS-depression (P = 0.005) and HADS-anxiety (P = 0.05). By contrast, in those with quiescent UC, those with an FGID did not have different median HADS scores (depression P = 0.15, anxiety P = 0.62) or IBD-QoL scores (P = 0.20), compared those without FGIDs. CONCLUSION: This study is one of the first to use Rome IV criteria in UC and confirms that the prevalence of FGIDs is high. Patients with active disease and overlapping functional symptoms appear to have worse QoL and more psychological distress compared to those with quiescent disease. Clinicians should be vigilant to this functional overlap and treat both functional and inflammatory driven symptoms.

Increased abundance of proteobacteria in aggressive Crohn\u2019s disease seven years after diagnosis

Intestinal dysbiosis in inflammatory bowel disease (IBD) patients depend on disease activity. We aimed to characterize the microbiota after 7 years of follow-up in an unselected cohort of IBD patients according to disease activity and disease severity. Fifty eight Crohn’s disease (CD) and 82 ulcerative colitis (UC) patients were included. Disease activity was assessed by the Harvey-Bradshaw Index for CD and Simple Clinical Colitis Activity Index for UC. Microbiota diversity was assessed by 16S rDNA MiSeq sequencing. In UC patients with active disease and in CD patients with aggressive disease the richness (number of OTUs, p = 0.018 and p = 0.013, respectively) and diversity (Shannons index, p = 0.017 and p = 0.023, respectively) were significantly decreased. In the active UC group there was a significant decrease in abundance of the phylum Firmicutes (p = 0.018). The same was found in CD patients with aggressive disease (p = 0.05) while the abundance of Proteobacteria phylum showed a significant increase (p = 0.03) in CD patients. We found a change in the microbial abundance in UC patients with active disease and in CD patients with aggressive disease. These results suggest that dysbiosis of the gut in IBD patients is not only related to current activity but also to the course of the disease.

Neutrophil/lymphocyte ratio and lymphocyte/monocyte ratio in ulcerative colitis as non-invasive biomarkers of disease activity and severity

BackgroundApart from endoscopic interventions, readily attainable cost-effective biomarkers for ulcerative colitis (UC) assessment are required. For this purpose, we evaluated differential leucocytic ratio, mainly neutrophil–lymphocyte ratio (NLR) and lymphocyte-monocyte ratio (LMR) as simple available indicators of disease activity in patients with ulcerative colitis.MethodsStudy conducted on 80 UC patients who were classified into two groups of 40 each according to Mayo score and colonoscopic findings. Group 1 (active UC) and group 2 (inactive UC). Another 40 group-matched healthy participants were enrolled. White blood cell count, NLR, LMR, C-reactive protein, and Erythrocyte sedimentation rate were measured and recorded.ResultsSignificant elevation of NLR was observed in active UC group compared to inactive UC and controls (2.63 ± 0.43, 1.64 ± 0.25, 1.44 ± 0.19 respectively; p < 0.0001). The optimal NLR cut-off value for active UC was > 1.91, with a sensitivity and a specificity of 90% and 90% respectively. The mean LMRs of active UC was significantly lower compared with inactive UC patients and controls (2.25 ± 0.51, 3.58 ± 0.76, 3.64 ± 0.49 respectively; p < 0.0001). The cut-off value of LMR for determining the disease activity was ≤ 2.88 with a sensitivity of 90% and a specificity of 90%. NLR, LMR, and CRP were found to be significant independent markers for discriminating disease activity (p = 0.000). Besides, NLR was significantly higher in patients with pancolitis and positively correlated with endoscopically severe disease.ConclusionNLRs and LMRs are simple non-invasive affordable independent markers of disease activity in UC.

Combined Signature of the Fecal Microbiome and Plasma Metabolome in Patients with Ulcerative Colitis.

BackgroundUlcerative colitis is a chronic, idiopathic inflammatory disease that destroys the colon structure. Nevertheless, the exact pathogenesis is not clear and needs to be fully elucidated.Material/MethodsStool and plasma samples were used for 16S ribosomal RNA sequencing and liquid chromatography mass spectrometry, respectively. In addition, we detected the level of trimethylamine N-oxide. Finally, we performed Pearson correlation analysis between the microbiome and the metabolome.ResultsTwenty-three active ulcerative colitis, 25 inactive ulcerative colitis, and 30 control cases were included. Thirty-four significantly different metabolites were found between the active ulcerative colitis and control groups, 38 were found between the inactive ulcerative colitis and control groups, and only 1 was found between the active ulcerative colitis and inactive ulcerative colitis groups. The plasma trimethylamine N-oxide level of the inactive ulcerative colitis and active ulcerative colitis groups was significantly higher than that of the control group. Moreover, we identified significant changes in 24, 18, and 12 bacterial genera for active ulcerative colitis-control, inactive ulcerative colitis-control, and active ulcerative colitis-inactive ulcerative colitis, respectively. Cross-correlation indicated an association between sphingosine 1-phosphate and Roseburia, Klebsiella, and Escherichia-Shigella. Through the pathway analysis, we found sphingolipid metabolism was one of the most significantly increased pathways.ConclusionsAlthough levels of trimethylamine N-oxide were higher in ulcerative colitis patients, they did not achieve statistical significance in active ulcerative colitis and inactive ulcerative colitis groups. Sphingosine 1-phosphate was increased in ulcerative colitis patients and there were several microbiota associated with it. Although further study is still needed, sphingosine 1-phosphate will probably become a new target for treatment of ulcerative colitis.

STUDY OF THE PROTEOMIC PROFILE IN PATIENTS WITH ULCERATIVE COLITIS, ITS CORRELATION WITH DIAGNOSIS AND DISEASE ACTIVITY

Background: Inflammatory bowel disease (IBD) comprises primarily 2 disorders: ulcerative colitis (UC) and Crohn's disease (CD). The hallmark of IBD is chronic, uncontrolled inflammation of the intestinal mucosa. The diagnosis is based on a combination of disease history, colonscopy, inflammatory biomarkers, radiological and histological evaluation. Most biomarkers used are not reliable and not disease specific, but reflect generalized inflammation. Aim: The aim of the work is to identify plasma proteomic profiles of UC cases and correlating this profile with the other diagnostic markers and activity of the disease. Methods: We performed a study with 70 plasma samples collected from patients classified in two groups (37 UC, 33 healthy controls) according to accredited criteria. They were subjected to: complete history taking, thorough clinical examination, laboratory investigations (erythrocyte sedimentation rate (ESR), C- reactive protein (CRP), fecal calprotectin, perinuclear antineutrophil cytoplasmic antibodies (P-ANCA), ileocolonscopy, histopathology, imaging. Plasma proteomic pattern of UC patients and control subjects was determined using Matrix-Assisted Laser Desorption/Ionization (MALDI) Time of Flight (TOF) Mass Spectrometer (MS) analysis. All plasma samples were subjected to solid-phase extraction (SPE). We analyzed the spectra obtained from all the samples using ClinProTool software. Results: There was a statistically significant difference of the plasma proteome profiles of UC group in comparison to health volunteers. 64 signals were identified by the ClinProTool software and of these 16 peptide peaks were highly significant (sensitivity was 100%, specificity was 84.4%, positive predictive value was 100, and negative predictive value was 82). There was a statistically significant difference between active versus inactive UC group, 5 integration regions used for classification between active and inactive UC patients using Genetic Algorithm model (GA) which gave 88.5% cross validation and 100% recognition capability. Markers as ESR, CRP, fecal calprotectin, P-ANCA are statistically correlated to the plasma proteomics found in UC patients. Conclusion: Proteomic profile has the potential to improve diagnosis and evaluate UC activity, reducing the need for more invasive techniques. The pattern of these peptides holds the promise of distinguishing disease states and providing clinically important information such as prognosis, response to therapy or perhaps targets of therapy.

Vascular endothelial growth factor, endostatin levels and clinical features among patients with ulcerative colitis and irritable bowel syndrome and among healthy controls: a cross-sectional analytical study.

Increased angiogenetic activity in inflammatory bowel disease (IBD) has been shown in previous studies. The aim of this study was to evaluate the relationship of serum vascular endothelial growth factor (VEGF) and endostatin levels with clinical features and mucosal expression in patients with ulcerative colitis (UC). Cross-sectional analytical study conducted in a tertiary-level public hospital. Serum VEGF and endostatin levels were determined in 82 individuals: 39 with UC, 28 with irritable bowel syndrome (IBS) and 15 healthy controls (HCs), using enzyme-linked immunosorbent assays (ELISA). VEGF and endostatin expressions were studied using immunohistochemistry (IHC). Mean serum VEGF and endostatin levels were significantly higher in patients with UC than in patients with IBS and in HCs (511.9 ± 377.5 pg/ml, 305.0 ± 121.42 pg/ml and 36.1 ± 40.6 pg/ml; P = 0.001 for VEGF; and 155.50 ± 59.8 ng/ml, 116.9 ± 23.8 ng/ml and 102.2 ± 22.4 ng/ml; P < 0.001 for endostatin, respectively). There was a positive correlation between serum VEGF and endostatin levels (r = 0.422; P < 0.01). Mean H-scores for VEGF expression were higher in the active UC group than in the inactive UC and IBS groups, in the stroma, endothelium and epithelium. Mean H-scores for endostatin expression were higher in the active UC group than in the inactive UC and IBS groups, in the stroma and endothelium. There was no endostatin expression in the epithelium. Increased endostatin appears to be a defensive reaction to increased VEGF in patients with UC.

Relationship between intestinal mucosa associated microbiota and clinical manifestations in the patients with ulcerative colitis

Objective To investigate the changes of the intestinal mucosa-associated microbiota in the patients with ulcerative colitis (UC), and to explore their correlation with the clinical manifestations. Methods From June to October 2016, at Gastrointestinal Endoscopy Center, the Second Affiliated Hospital of Xi′an Jiaotong University, 28 patients with UC and 16 healthy individuals who underwent colonoscopy examination were enrolled. The mucosa specimens of them were collected for fluorescent in situ hybridization (FISH). The bacterial flora were observed and counted, the correlation between the bacterial flora and the clinical manifestations were analyzed. Kruskal-Wallis test and Spearman rank correlation analysis were performed for statistical analysis. Results Among 28 patients with UC, 16 were at active phase and 12 at remission phase. The number of total bacteria flora, Escherichia coli, Clostridium and Bacteroides of the active UC group and remission UC group were all more than those of healthy control group; however the number of Lactobacillus and Bifidobacteria were less than those of healthy control group, and the differences were statistically significant (χ2=23.34, 19.94; 23.40, 12.96; 23.39, 19.16; 23.32, 10.46; 23.19, 4.25; 18.94, 12.33; all P<0.05). The number of total bacteria flora, Escherichia coli and Bacteroides of the active UC group were more than those of the remission UC group, however the number of Lactobacillus and Bifidobacteria were less than those of the remission UC group, and the differences were statistically significant (χ2=7.32, 5.63, 5.62, 20.38 and 4.82; all P<0.05). In the patients with UC, the defecation frequency was positively correlated with the count of Bacteroides (r=0.459, P=0.014) and was negatively correlated with the count of Lactobacillus (r=-0.634, P<0.01). In UC patients, bloody stool, endoscopic appearance and total Mayo score were positively correlated with the counts of universal bacteria, Escherichia coli and Bacteroides (r=0.469, 0.403, 0.376; 0.604, 0.562, 0.475; 0.551, 0.463, 0.461; all P <0.05); and which were negatively correlated with Lactobacillus and Bifidobacteria (r=-0.570, -0.413; -0.899, -0.458; -0.862, -0.480; all P<0.05). The evaluation by the doctors was positively correlated with the counts of Escherichia coli (r=0.415, P=0.028), however was negatively correlated with Lactobacillus and Bifidobacteri (r=-0.841, -0.529; both P<0.01). Conclusion The intestinal microbiota have sigificantly changed in patients with UC which are correlated with some clinical manifestations of UC. Key words: Colitis, ulcerative; In situ hybridization, fluorescence; Mucosa associated microbiota; Clinical manifestations

P166 Decreased expression of elafin in peripheral blood and intestinal mucosa in inflammatory bowel disease

Background: Elafin is an endogenic serine protease inhibitor that has diverse functions such as anti-inflammation, immunoregulation, anti-microbe and so on. The cause of inflammatory bowel disease (IBD) is still unclear. Recently, the destruction of balance between proteases and antiproteases become the focus in the pathogenisis of IBD. Several studies have reported the expression of elafin in colonic tissue of IBD patients or murine models, although the results are controversial, its therapeutic implication is noteworthy. Here, we detect the expression of elafin in peripheral blood and colonic tissue of IBD patients, then, explore its role and value in assessing the activity and severity of IBD. Methods: Real-time PCR and ELISA were respectively performed to quantitate elafin mRNA and plasma concentration in a total of 106 peripheral blood samples from 106 patients (41 UC, 27 CD, 38 controls). Compare the expression of elafin between groups and among each stage of IBD. Analyze the correlation between elafin and other markers (ESR, CRP, IBD activity score). Histological inflammation was scored, and elafin was localized and semiquantified by immunohistochemistry in a total of 49 colonic paraffin sections (17 inflamed, 13 noninflamed, 19 controls) from 36 patients (12 UC, 5 CD, 19 controls). Results: Elafin mRNA levels decreased significantly in active UC group, but increased in remission UC group. However, in CD, we did not detect the above signifcant differences. Moreover, elafin mRNA levels had tendency to decrease with increasing disease severity both in UC and CD. At the same time, elafin plasma concentration decreased both in UC and CD, but the differences among each stage were not signifcant. Extensive expression was detected in the colonic mucosa and gland cells in healthy controls, whereas in UC and CD, both in inflamed and noninflamed tissue, immunostaning of elafin weakened, especially in inflamed mucosa. Conclusions: The expression of elafin in peripheral blood and colonic inflamed tissue from active IBD patients decreases and correlates with the activity of IBD to some extent, suggesting that it may play a protective role in IBD and may predict disease activity and severity.

Correlation between neutrophils lymphocytes ratio and disease activity in patients with ulcerative colitis

Objective To investigate the correlation between neutrophils lymphocytes ratio (NLR) and disease activity of ulcerative colitis (UC). Methods Case-control study was used to compare NLR differences of 82 active UC patients, 45 inactive UC patients, and 254 healthy controls. The multivariate analysis was applied to analyze the correlation between the NLR, erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), white blood cells (WBCs), and UC activity. The sensitivity and specificity of NLR to identify UC activity was evaluated. Results NLR of active UC group was 2.45±1.22, which was significantly higher than that of inactive UC group and healthy control group, their NLR were 1.92±0.68 and 1.83±0.75, respectively (H=9.991, P=0.007). The multivariate analysis showed that only CRP was correlated with UC activity (OR=1.396, 95% CI: 1.086~1.795, P=0.009). Receiver operating characteristic (ROC) curve analysis showed that the optimum NLR cut-off point for UC activity was 2.23, the sensitivity and specificity were 55.82% and 62.75%, respectively. Conclusions NLR has a certain reference value for the evaluation of UC activity, but it cannot be independent as a clinical index to evaluate the UC activity. Key words: Colitis, ulcerative/IM; Neutrophils; Lymphocytes

Interleukin 27 is up-regulated in patients with active inflammatory bowel disease.

The aim of the study was to characterize and quantify tissue gene and protein expression of IL-27 in ulcerative colitis (UC) and Crohn's disease (CD) patients. This is an observational and cross-sectional study. Fifty-four patients with IBD were studied: 27 active UC, 12 inactive UC, 10 active CD, and 5 inactive CD. All patients belonged to the Inflammatory Bowel Disease Clinic at the Instituto Nacional de Ciencias Médicas y Nutrición. We found that IL-27 gene expression was significantly higher in active UC versus inactive UC group (P=0.015). The IL-27 mRNA expression was increased in patients with active CD compared with inactive CD disease (P=0.035). The percentage of IL-27 immunoreactive cells was higher in active UC versus active CD patients and non-inflamed tissue controls. The IL-27 was significantly elevated in active UC and CD patients, and it was associated with disease severity.

The relationship between the neutrophil–lymphocyte ratio and disease activity in patients with ulcerative colitis

Preliminary evidence suggests that a higher neutrophil–lymphocyte ratio (NLR) may be an indicator of active ulcerative colitis (UC). However, it is not clear whether the NLR is a useful and simple indicator of clinical activity in UC after adjusting for the other inflammatory markers. We designed a retrospective study to evaluate the role of the NLR in estimating disease severity in UC patients. The study consisted of 71 patients with UC and 140 age- and sex-matched healthy individuals (control group). The NLR, erythrocyte sedimentation rate, C-reactive protein, and white blood cell count were measured. The NLR values of the active UC group were elevated compared with those of the patients with inactive UC and the controls (2.59 ± 1.47, 2.03 ± 1.07, and 1.98 ± 0.85, respectively; p = 0.005). The receiver operating characteristic revealed that the optimum NLR cut-off point for active UC was 2.39. A multivariable logistic analysis showed that of the parameters studied, C-reactive protein was the only parameter able to significantly discriminate active from inactive UC (B: 0.222; p = 0.017; odds ratio: 1.248; 95% confidence interval: 1.041–1.497).

The changes of serum IFN-gamma, TNF alpha and IL-10 levels in ulcerative colitis and its relevance to helicobacter pylori infection

Objective To study the changes of serum IFN-gamma, TNF alpha and IL-10 levels in ulcera-tive colitis and its relevance to helicobacter pylori infection. Methods 80 UC patients were selected as the research object, and 50 cases of 80 patients were in the activity stage of UC,30 patients were in remission. In the 50 patients with active UC：14 cases were mild,26 cases were moderate, and 10 cases were severe.30 healthy subjects were set as healthy controls. Detection of serum IFN-gamma,TNF alpha, IL-10 levels and Hp infection were made. Results The serum TNF alpha and IL-10 levels of the activity group were higher than those of the remission group and the control group, the difference was statistically significant［TNF alpha:(54.75±5.92)ng／L vs.(23.69±4.15)ng／L vs.(20.45±4.01)ng／L,F＝36.79,P＝0.000;IL-10：（35.16±6.72)ng／L vs.(25.12±5.37)ng／L vs.(23.96± 5.21)ng／L,F＝35.68,P＝0.000］，the serum IFN-gamma level of the activity group was lower than those of the remission group and the control group, the difference were statistically significant［（14.98±3.15)ng／L vs.(26.06± 4.31)ng／L vs.(27.16±4.39)ng／L,F＝35.68,P＝0.000］. As the aggravating of patients with UC in activity stage, TNF alpha and IL-10 levels showed a trend of increase(F＝32.81,24.95,all P＜0.05), while IFN-gamma showed a trend of reduce(F＝22.29,P＜0.05).In 50 cases of active UC patients, 18 patients with Hp positive, 32 cases were Hp negative. In active UC group Hp positive patients'serum TNF alpha and IL-10 levels higher than the Hp negative patients, the difference was statistically significant(t＝4.62,4.35,all P＜0.01). Serum levels of IFN-gamma of active UC patients with Hp positive group was lower than the Hp negative patients, the difference was statistically significant(t＝5.91,P＜0.01). Conclusion In active UC patients expression of TNF alpha and IL-10 are high, and IFN-gamma expression is low. The expression of IFN-gamma, TNF alpha and IL-10 are closely related to Hp infection. Key words: Colitis，Ulcerative; Interferon gamma; Tumor necrosis factor alpha; Interleukin 10; Helicobacter pylori

Determination of the discriminant score of intestinal microbiota as a biomarker of disease activity in patients with ulcerative colitis

BackgroundIn recent years, the gut microbiota has been found to provide an important link to the development of inflammatory bowel diseases (IBD) like ulcerative colitis (UC). Accordingly, inter-individual variation in the gut microbial community may be linked to inter-individual variation in the risk of IBD or other diseases. Further, the Terminal Restriction Fragment Length Polymorphism (T-RFLP) is a molecular biology technique for profiling bacterial species in faecal samples. This study was to evaluate a biomarker based on intestinal microbiota.MethodsThe study subjects were 69 patients with UC together with 80 relatives as controls. Twenty-three patients had active UC (group I) and 46 had quiescent UC (group II). The later included 17 patients with mild inflammation in the large intestine (group IIa), 29 without inflammation (group IIb). The patients’ relatives were consanguineous (group III, n = 47), and non-consanguineous (group IV, n = 33). Faecal samples were obtained from all subjects for the investigation of intestinal microbiota by applying the T-RFLP method. The Discriminant analysis of operational-taxonomic-unit (OTU) on T-RFLP fingerprints was performed. The Canonical Discriminant Function Coefficient (Df) for each OTU was calculated. The individual OTUs were multiplied by the Df value, and the sum was termed the Discriminant Score (Ds).ResultsThe Ds decreased thus: group I > group IIa > group IIb > group III > group IV. Significant difference was calculated for group I vs group IV (P < 0.01), group I vs group IIb (P < 0.05), group I vs group III (P < 0.01), group IIa vs IV (P < 0.01), group IIb vs group IV (P < 0.01), group III vs group IV (P < 0.01), indicating a strong association between gut microbial species and the development of UC.ConclusionsIn this study, the Ds related to UC, or otherwise absence of UC in the five groups. Potentially, Ds may become a clinically relevant biomarker of disease activity in UC. To our knowledge, this is the first application of the Ds to the study of microbiota in UC patients, consanguineous and non-consanguineous relatives.Trial registrationClinical trial No: UMIN 000004123.

Peroxisome proliferator-activated receptors family is involved in the response to treatment and mild clinical course in patients with ulcerative colitis.

Background. PPARs play an important role in the regulation of intestinal inflammation. Methods. We included a total of 46 UC patients and 31 controls. The gene expression of PPARs was measured by RT-PCR and protein expression by immunohistochemistry. Results. PPARα gene expression was significantly decreased in patients with active UC compared with remission UC group (P = 0.001) and controls (P = 0.001). We found that low gene expression of PPARα in mucosa confers a higher risk of UC activity (P ≤ 0.0001, OR = 22.6). We observed an increase of PPARα expression in patients with UC who were treated with 5-aminosalicylates compared with those who received any other combined therapy (P = 0.03, OR = 0.08). PPARγ gene expression was decreased in the active UC group compared with UC in remission (P = 0.001) and control group (P = 0.001). An increased expression of PPARγ gene was associated with mild clinical course of the disease (P ≤ 0.001, OR = 0.05). No gene expression of PPARβ/δ was found in the colonic mucosa from UC patients and controls. Conclusion. Our results suggest that patients with high gene expression of PPARs have a better response to medical treatment and a mild clinical course of the disease.