NOD2/CARD15基因R702W、G908R及L1007fs多态性与广西壮族人群炎症性肠病的相关性

目的:探讨我国广西壮族人群NOD2/CARD15基因R702W、G908R及L1007fs的遗传多态性与炎症性肠病的相关性.方法:分别收集2007-02/2010-10在广西地区无亲缘关系的壮族(n=70)和汉族(n=76)IBD患者及壮族(n=80)和汉族(n=84)正常对照者的肠黏膜组织.采用酚氯仿法提取各组织样本DNA,采用聚合酶链反应-限制性片段长度多态性分析(PCR-RFLP)方法对NOD2/CARD15基因R702W、G908R及L1007fs进行检测,统计基因型及等位基因频率,分析上述3个多态性位点与广西壮族人群炎症性肠病的相关性.结果:广西壮族和汉族IBD患者与正常对照者均未发现NOD2/CARD15基因R702W、G908R及L1007fs突变型基因型,所有多态性位点上的基因型全部为野生型纯合子,其基因型频率和等位基因频率分布在IBD患者和正常对照者中差异无统计学意义(P>0.05).结论:NOD2/CARD15基因R702W、G908R及L1007fs多态性与广西壮族人群炎症性肠病无明显相关性.     

克罗恩病CARD15/NOD2基因突变的研究

目的 发现与中国人克罗恩病（CD）发病相关的单核苷酸多态性（SNP）及其与临床特点的关系.方法 临床资料完整的CD、溃疡性结肠炎（UC）患者及健康体检者各30例.提取人血白细胞基因组DNA,PCR扩增NOD2基因第4、8、11外显子,纯化后直接测序.结果 5例CD患者有SNP改变,其中2例为P268S,1例为R459R,2例为P268S和R459R,而在UC患者和健康人中各检测到1例R459R.所有研究对象未发现R702W、G908R及3020insC改变.CD有4例P268S,与UC和健康体检者比较差异有统计学意义（χ^2=8.037,P＜0.05）.4例P268S CD患者病变均在回肠（χ^2=9.231,P=0.01）,发病年龄小于20岁（χ^2=10.769,P＜0.01）,并发肠腔狭窄而需手术（χ^2=7.972,P＜0.01）,2例有P268S和R459R的患者病情较重,多次复发.结论 P268S可能是CARD15/NOD2基因中与中国人CD相关的SNP,与患者发病年龄、病变部位及肠腔狭窄明显相关,与患者性别及病变严重程度无关.     

NOD2/CARD15基因突变与中国人克罗恩病相关性的研究

背景：近年多项研究证明NOD2／CARD15基因序列的单核苷酸多态性（SNP）与西方白种人克罗恩病（CD）明显相关．其中3个SNP（R702W、G908R和3020ins C）与CD的相关性尤为显著。目的：探讨NOD2／CARD15基因SNP与中国人CD发病的相关性及其与CD临床特点的关系。方法：选取临床资料完整的CD患者48例、溃疡性结肠炎（UC）患者和健康对照者各50例，提取人血白细胞基因组DNA，经聚合酶链反应（PCR）扩增NOD2基因全部12对外显子，纯化后直接测序，根据结果分析其突变与CD病变特点的关系。结果：CD组、UC组和健康对照组均未检出3个西方人常见的NOD2／CARD15基因多态性位点。CD组的P268S突变率显著高于UC组和健康对照组（P〈0．05）。5例P268S突变的CD患者病变均位于回肠（P〈0．01），4例发病年龄≤20岁（P〈0．01），且均并发肠腔狭窄（P〈0．01）。结论：中国人CD患者中存在NOD2／CARD15基因P268S突变，且与患者的发病年龄、病变部位和并发症相关，有必要对其功能作进一步探讨。     

NOD2／CARDl5基因突变与中国人克罗恩病相关性研究

背景NOD2／CARDl5基因是人类克罗恩病（Crohn＇s disease，CD）第一个易感基因，既往研究发现P268S可能与中国人CD发病及临床特征相关。目的本研究旨在证实P268S与中国人CD发病及其临床特征的相关性。方法血样来自临床确诊的50例CD患者，60例溃疡性结肠炎（ulcerativecolitis，uc）患者及100例健康体检者（healthy controls，HC）。提取人血白细胞基因组DNA．PCR扩增目的片段，PCR—RFLP发现突变位点，DNA测序证实突变位点。结果共有8例CD患者发现有P268S改变，而在UC患者和HC中分别发现2例和3例P268S改变，CD组明显高于UC和HC组（X^2=10．829，P=0．004），而UC组和HC组无明显差异。8例有P268S改变的CD患者临床特征包括病变多位于回肠．发病年龄轻（6例〈20岁），常并发肠腔狭窄而需手术治疗，中一重度患者比例高。结论P268S可能是NOD2/CARDl5基因中与中国人CD相关的SNP。P268S与CD患者发病年龄、病变部位及并发症及病情严重程度可能相关。     

Clinical significance of NOD2/CARD15 and Toll-like receptor 4 gene single nucleotide polymorphisms in inflammatory bowel disease

AIM： To evaluate the role of genetic factors in the pathogenesis of Crohn＇s disease （CD） and ulcerative colitis （UC）, we investigated the single nucleotide polymorphisms （SNPs） of NOD2/CARD15 （R702W, Gg08R and L1007finsC）, and Toll-like receptor 4 （TLR4） genes （D299G and T399I） in a selected inflammatory bowel disease （IBD） population coming from Southern Italy. METHODS： Allele and genotype frequencies of NOD2/ CARD15 （R702W, Gg08R and L1007finsC） and TLR4 （D299G and T399I） SNPs were examined in 133 CD patients, in 45 UC patients, and in 103 healthy controls. A genotype-phenotype correlation was performed. RESULTS： NOD2/CARD15 R702W mutation was significantly more frequent in CD （9.8%） than in controls （2.4%, P = 0.001） and in UC （2.3%, P = 0.03）. No significant difference was found between UC patients and control group （P 〉 0.05）. In CD and UC patients, no significant association with G908R variant was found. L1007finsC SNP showed an association with CD （9.8%） compared with controls （2.9%, P = 0.002） and UC patients （2.3%, P = 0.01）. Moreover, in CD patients, G908R and L1007finsC mutations were significantly associated with different phenotypes compared to CD wild-type patients. No association of IBD with the TLR4 SNPs was found in either cohort （allele frequencies： D299G-controls 3.9%, CD 3.7%, UC 3.4%, P 〉 0.05; T399I-controls 2.9%, CD 3.0%, UC 3.4%, P 〉 0.05）. CONCLUSION： These findings confirm that, in our IBD patients selected from Southern Italy, the NOD2/ CARD15, but not TLR4 SNPs, are associated with increased risk of CD.     

CARD15/NOD2基因多态性与克罗恩病发病的荟萃分析

克罗恩病（CD）是一种具有遗传背景的多因素疾病。位于第16号染色体上的CARD15／NOD2基因被确定为克罗恩病的易感基因，该基因是参与细胞凋亡调控的Apaf21／PCed4超家族成员之一，主要在单核巨噬细胞内表达。有研究证明，CARD15／NOD2基因上三个多态位点（R702W、G908R、3020sinC）与克罗恩病发病相关。为综合评价CARD15／NOD2基因序列单核苷酸多态性同克罗恩病发病的相关性，我们用荟萃分析方法对二者的关系进行综合定量再分析，探讨CARD15／NOD2基因多态性与克罗恩病发病的关系。     

克罗恩病相关基因的研究进展

克罗恩病(Crohn’s disease,CD)是一种由多个基因共同参与的胃肠道慢性炎性肉芽肿性疾病。本文对近年来CD的相关基因NOD2/CARD15、ATG16L1、IRGM、IL-23/TH17信号通路、PTGER4、IBD5基因位点、PTPN2的研究进展进行总结。通过检索PUBMED及CNKI数据库,以"克罗恩病、相关的基因、发病机制"为关键词,以CD的相关的基因和其致病机制、相关基因在CD中的作用及临床意义、CD相关基因位点临床治疗效果为纳入标准,检索了2000年1月至2017年8月的相关文献。发现NOD2/CARD15、ATG16L1、IRGM、IL-23/TH17信号通路、PTGER4、IBD5基因位点、PTPN2等基因与CD的遗传易感性密切相关。这些基因与CD的临床表现、治疗效果和患者生存率等相关,为CD今后发病机制的研究,及相关基因靶向的治疗奠定了理论基础。     

中国汉族人群NOD2、IRGM、ATG16L1和STAT4基因多态性与克罗恩病的相关性研究

背景：克罗恩病（CD）的病因和发病机制尚未完全阐明,近年国外研究发现NOD2、IRGM、ATG16L1、STAT4基因突变与CD相关。目的：分析NOD2、IRGM、ATG16L1、STAT4基因多态性与中国汉族人群CD发病的相关性。方法：连续纳入2007年1月～2010年1月苏州市立医院中国汉族CD患者66例,66名健康体检者作为正常对照,以PCR联合基因测序检测4种基因相应单核苷酸多态性（SNP）位点的基因型,分析各基因型和等位基因频率。结果：CD组和正常对照组NOD2基因rs2066842位点、IRGM基因rs13361189位点、ATG16L1基因rs2241880位点和STAT4基因rs7574865位点基因型和等位基因频率分布均符合Hardy-Weinberg遗传平衡定律,两组间4种基因相应SNP位点的基因型和等位基因频率差异均无统计学意义。结论：NOD2、IRGM、ATG16L1和STAT4基因多态性与中国汉族人群CD发病不相关。     

NOD2/CARD15基因突变与中国人克罗恩病相关性研究

背景 NOD21CARDl5 基因是人类克罗恩病(Crohn's disease,CD)第一个易感基因,既往研究发现P268S可能与中国人CD发病及临床特征相关.目的 本研究旨在证实P268S与中国人CD发病及其临床特征的相关性.方法 血样来自临床确诊的50例CD患者,60例溃疡性结肠炎(ulcerative colitis,uc) 患者及100 例健康体检者(healthy controls,HC).提取人血白细胞基因组DNA,PCR 扩增目的片段,PCR-RFLP发现突变位点,DNA 测序证实突变位点.结果 共有8例CD患者发现有P268S改变,而在UC患者和 HC 中分别发现2例和3例P268S改变,CD组明显高于UC和HC组(x2=10.829,P=0.004),而UC组和HC组无明显差异.8例有P268S改变的CD患者临床特征包括病变多位于回肠.发病年龄轻(6例<20岁),常并发肠腔狭窄而需手术治疗,中一重度患者比例高.结论 P268S可能是NOD21CARDl5 基因中与中国人CD相关的SNP.P268S与CD患者发病年龄、病变部位及并发症及病情严重程度可能相关.     

NOD2/CARD15基因多态性与克罗恩病患者相关性研究

目的NOD2/CARD15基因是人类的第一个克罗恩病(CD)易感基因,其间的3个单核苷酸多态性(SNPs)与白种人CD有显著性相关,但与日本人无关。本研究旨在证实这3个SNPs是否与浙江地区人群的CD易感性有关。方法血样来自浙江地区32例CD患者,110例溃疡性结肠炎患者及292例健康对照者。通过PCRSSP方法直接检测野生型及NOD2/CARD15基因的3个多态性(Arg702Trp,Gly908Arg,Leu1007fsinsC)。结果没有发现1例CD患者纯合子或杂合子的SNPs突变,同样在溃疡性结肠炎患者和健康人中也未能检测到。结论本研究表明一些存在于特定人群的CD易感基因可能在其他人群中不存在,白种人CD患者相关的易感基因NOD2/CARD15常见的3个SNPs与浙江地区CD人群无关。     

P268S突变型NOD2/CARD15真核表达载体的构建及其体外表达

背景：NOD2/CARD15基因序列单核苷酸多态性（SNP）与欧美人群的克罗恩病（CD）明显相关．其中R702W、G908R和3020insC3个SNP位点与CD的相关性尤为显著。而13本、韩国以及我国香港和浙江地区的研究均未发现上述3个SNP的改变，但最近研究发现了可能与中国人CD相关的P268S突变。目的：构建P268S突变型NOD2／CARD15真核表达载体和体外转染体系，为研究突变型NOD2／CARD15的功能提供实验基础。方法：应用定点诱变技术构建P268S突变型NOD2／CARD15真核表达载体，以阳离子脂质体介导体外转染技术瞬时转染人胚肾细胞HEK293T．以蛋白质印迹法和逆转录聚合酶链反应（RT—PCR）检测HEK293T细胞NOD2／CARD15的表达。结果：经克隆、酶切、测序证实获得P268S突变型NOD2／CARD15基因，突变载体转入HEK293T细胞后，NOD2／CARD15有效表达。结论：成功构建了P268S突变型NOD2／CARD15真核表达载体，阳离子脂质体是人胚肾细胞有效的体外转染体系。     

NOD2/CARD15基因突变与克罗恩病的相关性研究进展

克罗恩病(CD)是炎症性肠病(IBD)的一种，其病因及发病机制仍不明确，大量证据表明是由遗传因素、环境因素自身免疫紊乱三者相互作用的结果，其中遗传易感因素在CD发病机制中起重要作用。其遗传易感性主要表现为家族聚集现象、单卵双生共患率及不同人群发病率、流行率差异较大。自2001年发病并证实人类CD第一个易感基因NOD2／CARD15以来，关于该基因突变与CD相关性研究已成为IBD研究领域的热点，本文就其研究进展作一综述。     

NOD2/CARD15与克罗恩病相关性研究进展

克罗恩病(CD)是一种常见的炎症性肠病,是环境因素作用于易感个体产生的肠道慢性非特异性炎症。其发病机制尚未完全明了。NOD2/CARD15是目前确认的第一个与CD相关的基因,通过介导凋亡,调节免疫等多种途径参与CD的发病。本文就NOD2/ CARD15与CD的研究进展作一综述。     

自噬相关遗传变异与克罗恩病

克罗恩病（CD）是一种肠道慢性非特异性炎性疾病,其发病机制尚未完全明确。目前普遍认为CD是由于环境因素作用于基因突变的易感个体而发病。目前已发现30多个基因位点的突变与CD相关,包括NOD2、TLR和ATG16L1,这3个基因均直接或间接与自噬过程相关。此文就自噬与CD发病关系作一综述,为临床寻找新的CD治疗方法提供依据。     

NOD2,IL23R and ATG16L1 polymorphisms in Lithuanian patients with inflammatory bowel disease

AIM:To investigate the frequency of NOD2, IL23R and ATG16L1 genetic variants in a case-control panel for inflammatory bowel disease (IBD) from Lithuania.METHODS: One hundred and eighty unrelated IBD pa- tients [57 Crohn‘s disease (CD) and 123 ulcerative colitis (UC)] and 186 healthy controls were genotyped for the following known genetic susceptibility variants:NOD2-Arg702Trp (rs2066844), Gly908Arg (rs2066845) and Leu1007insC (rs2066847), as well as IL23R-Arg381Gln (rs11209026) and ATG16L1-Thr300Ala (rs2241...     

CD第一个易感基因NOD2及其与CD易感性的研究进展

目前认为克罗恩病(CD)是遗传易感受环境因素作用而发病，其发病符合多基因病的遗传规律^[1]，其遗传易感性主要表现在家族聚集现象及双胞胎共患率上^[2]。近年来，随着人类基因组计划的进展和多基因病研究及统计学方法的发展，已发现了人类CD的第一个易感基因—NOD2基因，现命名为CARDl5基因，本就NOD2基因及其与CD易感性的研究进展作一综述。     

Crohn��s disease genotypes of patients in remission vs relapses after infliximab discontinuation

AIM:To investigate genetic differences between Crohn's disease(CD) patients with a sustained remission vs relapsers after discontinuing infliximab while in corticosteroid-free remission.METHODS:Forty-eight CD patients received infliximab and were in full corticosteroid-free clinical remission but then discontinued infliximab for reasons other than a loss of response,were identified by review of an electronic database and charts.Infliximab-associated remission was defined as corticosteroid-free plus normalization of clinical disease activity [CD activity index(CDAI) 150] during follow-up visits based on physician global assessments.A CD relapse(loss of infliximab-induced remission) was clinically defined as a physician visit for symptoms of disease activity(CDAI 220) and a therapeutic intervention with CD medication(s),or a hospitalization with complications related to active CD.Genetic analyses were performed on samples from 14 patients(n = 6 who had a sustained long term remission after stopping infliximab,n = 8 who rapidly relapsed after stopping infliximab).Nucleotide-binding oligomerization domain 2(NOD2)/caspase activation recruitment domain 15(CARD15) polymorphisms(R702W,G908R and L1007fs) and the inflammatory bowel disease 5(IBD5) polymorphisms(IGR2060a1 and IGR3081a1) were analyzed in each group.RESULTS:Five single nucleotide polymorphisms of IBD5 and NOD2/CARD15 genes were successfully analyzed for all 14 subjects.There was no significant increase in frequency of the NOD2/CARD15 polymorphisms(R702W,G908R and L1007fs) and the IBD5 polymorphisms(IGR2060a1 and IGR3081a1) in either group of patients;those whose disease relapsed rapidly or those who remained in sustained long term remission following the discontinuation of infliximab.Nearly a third of patients in full clinical remission who stopped infliximab for reasons other than loss of response remained in sustained clinical remission,while two-thirds relapsed rapidly.There was a marked difference in the duration of clinical remission following discontinuance of infliximab between the two groups.The patients who lost remission did so after 1.0 years ± 0.6 years,while those still in remission were at the time of this study,8.1 years ± 2.6 years post-discontinuation of infliximab,P 0.001.The 8 patients who had lost remission after discontinuing infliximab had a mean number of 5 infusions(range 3-7),with a mean treatment time of 7.2 mo(range 1.5 mo-15 mo).The mean duration of time from the last infusion of infliximab to the time of loss of remission was 382 d(range 20 d-701 d).The 6 patients who remained in remission after discontinuing infliximab had a mean number of 6 infusions(range 3-12),with a mean treatment duration of 12 mo(range 3.6 mo-32 mo)(P = 0.45 relative to those who lost remission).CONCLUSION:There are no IBD5 or NOD2/CARD15 mutations that predict which patients might have sustained remission and which will relapse rapidly after stopping infliximab.     

NOD2基因P268S突变是中国人克罗恩病的易感基因吗?

本期龙靖华等作者发表的论文，采用聚合酶链反应（PCR）扩增NOD2／CARD15基因12对外显子．纯化后直接测序的方法，研究了我国广东地区48例克罗恩病（CD）患者、50例溃疡性结肠炎（UC）患者和50名健康对照者NOD2基因突变与炎症性肠病（IBD）的相关性，结果发现了一个有意义的现象。CD组NOD2P268S突变率显著高于UC组和健康对照组．且与CD患者的发病年龄、     

Altered pattern of tumor necrosis factor-alpha production in peripheral blood monocytes from Crohn's disease

AIM To evaluate the inflammatory state in Crohn's disease(CD) patients and correlate it with genetic background and microbial spreading.METHODS By means of flow cytometry, production of tumor necrosis factor-alpha(TNF-α) was measured in peripheral blood monocytes from patients suffering from CD, ulcerative colitis(UC) and in healthy subjects after stimulation of the NOD2 and TLR pathways. CD patients were genotyped for the three most common NOD2 variants(R702W, G908 R and L1007Pfs*2) and basal production of TNF-α was correlated to NOD2 genotype. Also, production of TNF-α was correlated to plasmatic levels of LPS Binding Protein(LBP), soluble(s) CD14 and to the activity state of the disease.RESULTS The patients with CD were characterized by a significantly higher monocyte basal expression of TNF-αcompared with healthy subjects and UC patients, and after stimulation with Pam3CSK4(ligand of TLR2/1) and MDP-L18(ligand of NOD2) this difference was maintained, while other microbial stimuli(LPS, ligand of TLR4 and Poly I:C, ligand of TLR3) induced massive activation in CD monocytes as well as in UC and in healthy control cells. There was no significant difference in the production of TNF- α between patients who carried CD-associated heterozygous or homozygous variants in NOD2 and patients with wild type NOD2 genotype. Although serum LBP levels have been shown to correlate positively with the state of activity of the disease, TNF-α production did not show a clear correlation with either LBP or s CD14 levels in plasma. Moreover, no clear correlation was seen between TNF-α production and activity indices in either CD or UC.CONCLUSION Peripheral monocytes from CD express higher basal and stimulated TNF-α than controls, regardless of NOD2 genotype and without a clear correlation with disease activity.     

冠心病患者血小板表面Fc受体γ链ⅡA和CD40配体的表达

目的研究冠心病患者血小板表面Fc受体γ链ⅡA(FcγRⅡA)和CD40配体(CD40L)的表达水平及其与冠状动脉(冠脉)粥样硬化病变程度的相关性。方法采用流式细胞术检测健康对照组(30名)及冠心病患者(95例)血小板表面FcγRⅡA和CD40L的表达水平,并将冠心病组上述指标与该组患者冠脉造影病变支数Gensini积分进行相关性分析。结果与对照组比较,冠心病组血小板表面FcγRⅡA和CD40L的表达水平叫显升高(P<0.01),血小板FcγRⅡA的表达水平(r_1=0.45,r_2=0.51,r_3=0.57,P<0.05)及CIM01.的表达水平(r_1=0.52,r_2=0.55,r_3=0.63,P<0.05)与冠脉病变支数Gensini积分均呈正相关。结论血小板表面FcγRⅡA和CD40L的表达水平在冠心病患者中升高,可作为评估冠脉病变严重程度的指标。