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Objective:To analyze the influence of vitamin D 3 supplementation on the clinical efficacy of mesalazine in patients with ulcerative colitis (UC). Methods:From January 2015 to December 2020, patients with mild-to-moderate active UC were retrospectively and continuously enrolled, who accepted mesalazine treatment for at least 12 months at the Second Affiliated Hospital of Wenzhou Medical University. According to simultaneous supplement of vitamin D 3 (125 IU/d), the patients were divided into study group and control group. Demographic and disease characteristics, serum 25-hydroxyvitamin D[25(OH)D] levels and other information were collected through retrieving hospital database. Student′s t-test, Mann-Whitney U test and Chi-square test were applied for comparison of disease characteristics. The changes of modified Mayo scores[ΔMayo] and 25(OH)D[Δ25(OH)D] were compared before and after treatment by paired t-test, Wilcoxon signed rank test and Chi-square test. Multiple linear regression model was used to analyze the independent factors affecting ΔMayo and Δ25(OH)D, and variables with P-values less than 0.20 in the univariate analysis were allowed for further multivariate analysis. Results:A total of 74 UC patients (44 males, 30 females), with median age (range) 39.5 (20-76) years old, were analyzed and respectively assigned into study group ( n=36) and control group ( n=38). In study group, the average level of serum 25(OH)D was significantly increased at month 12 compared with that at baseline [(22.87±7.30) μg/L vs. (18.15±7.48) μg/L, P<0.001]. However, no significant elevation of serum 25(OH)D was found in control group [(19.17±8.49) μg/L vs. (19.82±9.47) μg/L, P=0.466]. Furthermore, there was a significant decrease of modified Mayo score [-3(-4.75, -1.25) vs.-2(-3.25, 0), P=0.034] and a higher clinical remission rate (55.6% vs. 28.9%, P=0.020) at month 12 in study group than those in control group. In addition, according to the baseline level of serum 25(OH)D before mesalazine treatment, 74 UC patients were divided into vitamin D deficiency group ( n=38, serum 25(OH)D<20 μg/L) and non-deficiency group ( n=36, serum 25(OH)D≥20 μg/L). At month 12 in vitamin D deficiency group, patients with vitamin D3 supplementation had a greater decline in modified Mayo score [-4(-5.75, -2) vs.-2(-4, 0), P=0.048] and a higher clinical remission rate (60.0% vs. 22.2%, P=0.019) compared with those without. Conclusions:In patients with mild-to-moderate active UC receiving mesalazine treatment, vitamin D3 supplementation may improve the clinical efficacy, especially in patients with vitamin D deficiency.
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Background: Imbalance of Th17/Treg cells might play a key role in the initiation of Crohn's disease (CD). Moreover, tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) pathway was suggested to be implicated in the pathogenesis of CD and other autoimmune diseases. Aims: To investigate the influence of TRAIL gene deletion on the balance of Th17 and Treg cells in mice with experimental colitis. Methods: TRAIL-/- C57BL/6 mice were obtained by CRISPR/Cas9 method. Then 20 TRAIL-/- mice and 20 wild-type (WT) mice were randomly divided into WT group, TRAIL-/- group, WT trinitrobenzenesulfonic acid (TNBS)-induced colitis group and TRAIL-/- TNBS-induced colitis group, with 10 mice in each group. The severity of colonic inflammation was assessed by disease activity index (DAI) and histology activity index (HAI). Peripheral blood and colonic tissue were collected to determine the expression levels of Th17 and Treg cell-associated transcription factors (RORγt, Foxp3) and cytokines [interleukin-17 (IL-17), IL-10] by real-time PCR, Western blotting and ELISA method, respectively. Results: Compared with untreated mice, mice with experimental colitis showed decreased body weight, shortened colon length as well as increased DAI and HAI (P<0.05), and the manifestations of colitis in TRAIL-/- mice were more serious than that in WT mice (P<0.05). In WT mice with experimental colitis, the expression levels of RORγt and IL-17 in peripheral blood and colonic tissue were significantly increased than those in untreated WT mice (P<0.05), while the expression levels of Foxp3 and IL-10 were significantly decreased (P<0.05). Furthermore, the expression levels of all four Th17 and Treg cell-associated molecules were higher in TRAIL-/- colitis mice than in WT colitis mice (P<0.05). In addition, the ratio of RORγt to Foxp3 were higher in TRAIL-/- colitis mice than in WT colitis mice (P<0.05). Conclusions: Deletion of TRAIL gene may aggravate the severity of colonic inflammation via up-regulating Th17/Treg ratio in mice with experimental colitis.
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Objective To investigate the relationship between forkhead/winged helix transcription factor (Foxp) 3 gene polymorphisms and susceptibility and phenotype of Crohn's disease (CD) in Han nationality in Zhejiang province.Methods From January 2007 to December 2015,268 diagnosed CD patients and 490 healthy controls were enrolled.The four single nucleotide polymorphism (SNP) of Foxp3 rs3761547,rs2232365,rs2294021 and rs3761548 were examined by a SNaPshot technique,and their relation with the efficacy of infliximab was evaluated.The linkage disequilibrium (LD) and haplotype were also analyzed.Unconditional Logistic regression analysis was performed for statistical analysis.Results There was no significant difference in the four mutant alleles and genotype frequencies between 31 patients with effective infliximab treatment and 19 patients with ineffective treatment (all P＞0.05).The results of LD analysis indicated that the above four SNP were in a tight linkage.The frequency of haplotype GCGC of male CD group was 29.20％ (40/137),which was higher than that of male healthy control group (19.37％,43/222),and the difference was statistically significant (odd ratio (OR)=1.717,95％ confidence interval (CI) 1.045 to 2.820,P=0.032).The frequency of haplotype ACGA of female CD group was 13.36％ (35/262),which was lower than that of female healthy control group (19.03％,102/536),and the difference was statistically significant (OR=0.656,95％CI 0.433 to 0.995,P=0.046).The frequency of haplotype ATAC of male colon (L2) type was 25.93％ (7/27),which was lower than that of ileocecal colon (L3) type (75.38％,49/65),and the difference was statistically significant (OR=0.114,95％CI 0.041 to 0.320,P＜0.01).The frequency of haplotype GCGC of male L2 type was 51.85％ (14/27),which was higher than that of L3 type (9.23％,6/65),and the difference was statistically significant (OR=10.590,95％CI 3.423 to 32.758,P＜0.01).The frequency of haplotype ATAC of male stenotic (B2) type was 73.21％ (41/56),which was higher than that of nonstenotic and nonpenetrated (B1) type (47.30％,35/74),and the difference was statistically significant (OR=0.328,95％CI 0.156 to 0.693,P=0.003).The frequency of haplotype GCGC of male B2 type was 17.86％ (10/56) which was lower than that of nonstenotic and nonpenetrated (B1) type (39.19％,29/74),and the difference was statistically significant (OR=2.946,95％CI 1.295 to 6.784,P=0.009).The frequency of haplotype ACGA of male penetrated (B3) type was 71.43％ (5/7),which was higher than that of nonstenotic and nonpenetrated (B1) type (12.16％,9/74),and the difference was statistically significant (OR =0.055,95％ CI 0.009 to 0.329,P ＜ 0.01).Conclusion Foxp3 (rs3761547,rs2232365,rs2294021,rs3761548) gene polymorphisms are associated with the susceptibility and phenotype of CD in Chinese Han patients,but not related with the efficacy of infliximab.
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<p><b>OBJECTIVE</b>To assess the association of vascular endothelial growth factor (VEGF) gene polymorphisms with susceptibility to Crohn's disease (CD) in a Chinese population.</p><p><b>METHODS</b>For 275 CD patients and 495 controls, the genotypes of VEGF gene rs699947 and rs3025039 loci were determined with a SNaPshot method.</p><p><b>RESULTS</b>The allelic and genotypic frequencies of the rs699947 and rs3025039 loci did not differ between the two groups (all P>0.05). By stratification analysis, allele A and genotype CA+AA of rs699947 were more frequent in patients with colonic CD compared with the controls (P=0.006, 95%CI:1.143-2.234; P=0.005, 95%CI:1.203-2.900, respectively). Compared with the controls, the allele A and genotype CA+AA of rs699947 were less frequent in patients with ileal lesions including ileal CD and ileocolonic CD (P=0.033, 95%CI:0.524-0.974;P=0.043, 95%CI:0.481-0.989, respectively). The frequency of TT homozygote of rs3025039 was lower in patients with non-stricturing and non-penetrating CD compared with the controls (P=0.036, 95%CI:0.016-0.870).</p><p><b>CONCLUSION</b>Polymorphisms of the VEGF gene rs699947 locus may contribute to an increased risk for colonic CD, but may play a protective role in patients with ileal lesion. Individuals carrying the TT genotype for VEGF rs3025039 locus may be less susceptible to non-stricturing and non-penetrating CD.</p>
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Objective To explore the relationship of Crohn's disease (CD) susceptibility to aryl hydrocarbon receptor (AhR) polymorphisms and haplotypes in Han population in Wenzhou city, China. Methods A total of 310 CD patients and 573 age-and sex-matched healthy controls were enrolled in our study. Three single nucleotide polymorphisms (SNPs) of AhR(rs10249788,rs2158041,rs2066853) were determined by the improved multiple ligase detection reaction technique. Unconditional logistic regression analyses was applied to analyze the allelic and genotypic differences of each SNP between CD patients and controls, as well as their influence on the clinicopathologic characteristics in CD patients. Analyses of linkage disequilibrium and haplotype were performed by Haploview 4.2 software in all study subjects. Results Compared with the controls, the variant allele (T) and genotype (CT+TT) of (rs2158041) were evidently decreased among CD patients (19.52% vs. 25.04%, P=0.009; 34.19% vs. 44.68%, P=0.003). According to"the Montreal Classification Standards", CD patients were divided into different subgroups. The variant allele(T)and genotype(CT+TT)of(rs2158041)were significantly lower in patients with terminal ileum CD than in controls (16.79% vs. 25.04%, P=0.005; 28.24% vs. 44.68%, P=0.001). Similar conclusions were also drawn in patients with constricting disease when compared with the controls(15.20%vs.25.04%,P=0.003;28.43% vs.44.68%,P=0.003).The three SNPs above were shown to be in a linkage disequilibrium.Compared with the controls respectively,the frequency of haplotype(CCG)was increased in CD patients (44.73% vs. 39.60%, P=0.039), whereas that of haplotype (CTG) was decreased (18.02% vs. 22.78%, P=0.047). Conclusions AhR (rs2158041) variation might influence the risk as well as the location and behavior of CD. The haplotype (CCG) possibly increase the risk of CD development, whereas haplotype(CTG)might decrease it.
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Dysregulated immune response is crucial for the pathogenesis of inflammatory bowel disease.Recent studies suggested that imbalance among Th1, Th2 and Th17 cells was closely related to the aberrant intestinal immune response.Aims: To investigate the association of imbalance among Th1, Th2 and Th17 cells and Crohn''s disease (CD).Methods: Thirty-six CD patients admitted from Jan.2013 to Dec.2014 at the Second Affiliated Hospital of Wenzhou Medical University were enrolled;40 patients undergoing intestinal polypectomy were collected as controls.Inflamed and normal mucosal tissues were obtained from CD patients and the controls, respectively;expressions of Th1, Th2 and Th17 cells related transcriptional factors (T-bet, GATA-3 and RORγt) and cytokines (IFN-γ, IL-4 and IL-17A) were determined by real-time PCR and immunohistochemistry.Results: In comparison with the controls, mRNA expression of T-bet and RORγt, mRNA and protein expressions of IFN-γ and IL-17A, as well as the ratios for T-bet/GATA-3 and RORγt/GATA-3, which represented balance of Th1/Th2 and Th17/Th2, respectively, were all significantly increased in inflamed mucosal tissue in CD patients (P<0.05).Expression of each of the three transcriptional factors was positively correlated with its corresponding cytokine (P<0.05).IFN-γ+ and IL-17A+ cells mainly located in the intestinal epithelial layer and lamina propria with cytoplasmic immunoreactivities in CD patients.In the stratified analysis, all the above-mentioned parameters were significantly higher in active CD than in inactive CD (P<0.05);furthermore, the ratio for RORγt/T-bet, which represented balance of Th17/Th1, was also increased significantly in active CD (P<0.05).Conclusions: Imbalance among Th1, Th2 and Th17 cells in intestinal mucosal tissue was closely related with CD.Polarization of Th1 and Th17 cells are involved in this process, and Th17 polarization is predominant in active disease.
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Objective To explore the relation between genetic polymorphisms and the expression in colonic tissues of solute-linked carrier family 26 member A3 (SLC26A3) and susceptibility of Crohn's disease (CD) in Han population of Zhejiang Province.Methods A total of 265 CD patients and 566 gender-and age-matched healthy individuals were enrolled.Alleles and genotypes of SLC26A3 (rs17154444,rs7810937,rs7785539,rs2108225,rs6951457) were examined by SNaPshot.The linkage disequilibrium (LD) and haplotype were also analyzed.Eight patients with colonic CD and eight genderand age-matched patients with benign colonic polyps (control group) were selected.The expression level of SLC26A3 protein in the colonic tissue was detected by immunohistochemistry.T test and rank-sum test were performed for statistical analysis.Unconditional Logistic regression analysis was used to analyze the distributions of SLC26A3 polymorphisms and their effects on the clinicopathological features of CD patients.Results The frequencies of mutant allele of rs2108225,rs7785539 and rs6951457 of the CD group were 53.77％ (285/530),4.72％ (25/530) and 2.83％ (15/530),and the frequencies of mutant genotype were 76.23 ％ (202/265),9.43 ％ (25/265) and 5.66 ％ (15/265),which were lower than those of the control group (60.95％,690/1 132;8.13％,92/1 132;6.10％,69/1 132;83.92％,475/566;15.37％,87/566 and 11.84％,67/566),and the differences were statistically significant (all P＜0.05).The frequencies of mutant allele of rs17154444 and rs7810937 of the CD group were 10.19％ (54/530) and 34.91 ％ (185/530),and the frequencies of mutant genotype were 18.49 ％ (49/265) and 56.23 ％ (149/ 265),compared with those of the control group (8.30％,94/1 132;30.92％,350/1 132;15.55％,88/566 and 51.77％,293/566),the differences were not statistically significant (all P＞0.05).The frequency of mutant allele G of rs2108225 in patients with ileal CD was 47.89 ％ (91/190),and the frequency of mutant genotypeAG+GG was 65.26％(62/95),which were both lower than those of colonic CD (61.62％,122/198 and 85.86％,85/99),and the differences were statistically significant (both P＜0.012 5).rs7810937,rs7785539 and rs2108225 were in a strong linkage disequilibrium.The frequencies of haplotypes AGG and ACA of the CD group were 53.96％ (286/530) and 4.34％ (23/530),which were lower than those of the control group (60.07％,680/1 132 and 7.51％,85/1 132),and the differences were statistically significant (52 =5.534,P=0.019;x2 =5.967,P=0.015).And the frequency of haplotype AGA of the CD group was 8.30％ (44/530),which was higher than that of the control group (1.15％,13/1 132),and the difference was statistically significant (x2 =7.793,P＜0.01).Furthermore,the expression level of SLC26A3 protein in colonic tissues of eight colonic CD patients was 0.19±0.07,which was lower than that of patients with benign colonic polyps (0.26 ±-0.03),and the difference was statistically significant (t=2.55,P=0.023).In addition,the expression levels of SLC26A3 protein in patients carrying genotype GG or AG of rs2108225 were 0.19±0.03 and 0.10±0.01,respectively,which were lower than that of patients carrying genotype AA (0.26± 0.02),and the differences were statistically significant (t=3.19,P=0.033;t=9.06,P=0.003).Conclusions The genetic polymorphismns and their haplotypes of SLC26A3 (rs7785539,rs2108225 and rs6951457) are associated with the susceptibility of CD,and SLC26A3 (rs2108225) polymorphism may affect the expression level of SLC26A3 protein in the colonic tissues.
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<p><b>OBJECTIVE</b>To assess the association of transcobalamine II (TCN2) gene polymorphisms and serum levels of homocysteine (Hcy), vitamin Band folate with ulcerative colitis (UC) among Chinese patients.</p><p><b>METHODS</b>For 397 UC patients and 574 controls, two single nucleotide polymorphisms of the TCN2 gene (rs1801198, rs9606756) were tested with an improved multiple ligase detection reaction method. Serum Hcy, vitamin Band folate were measured with an enzymatic cycling assay and an chemiluminescence immunoassay, respectively.</p><p><b>RESULTS</b>The allelic and genotypic frequencies of rs1801198 and rs9606756 did not differ significantly between the two groups (all P> 0.05). Compared with those of the control group, the frequencies of G allele and CG+GG genotype of rs1801198 were greater in patients with moderate and severe UC (both P< 0.05). The same conclusion may also be drawn for the G allele and AG genotype of rs9606756 (both P< 0.05). Compared with the controls, average Hcy level was enhanced in UC patients (P< 0.01), whereas average vitamin Band folate levels were decreased in UC patients (both P< 0.01). In both groups, the average level of Hcy was lower in individuals carrying CC of (rs1801198) than in those with CG+GG (both P< 0.05). A similar conclusion was also drawn for individuals with AA of rs9606756 when compared with those carrying AG(both P< 0.05). Compared with patients with mild UC, average Hcy level was increased in those with moderate and severe UC (P< 0.01), while average vitamin Band folate levels were decreased in those with moderate and severe UC (both P< 0.01). The prevalence of hyperhomocysteinemia(HHcy), vitamin Bdeficiency and folate deficiency was greater in UC patients than in controls (all P< 0.01). In UC patients, the level of Hcy was negatively correlated with those of vitamin B(P< 0.01), albumin(P< 0.01), red blood cells(P< 0.01) and platelet (P< 0.05), but positively correlated with white blood cells(P< 0.01) and Mayo score (P< 0.01). Both HHcy and folate deficiency were independent risk factors for UC (OR=4.173, OR=5.206, both P< 0.01).</p><p><b>CONCLUSION</b>TCN2 (rs1801198, rs9606756) variations, as well as serum levels of Hcy, vitamin Band folate, are correlated with UC. Both HHcy and folate deficiency are independent risk factors for UC.</p>