类风湿关节炎患者外周血单个核细胞中PADI4 mRNA的表达及其意义

目的检测类风湿关节炎(RA)患者外周血单个核细胞(PBMCs)肽酰基精氨酸脱亚氨酶4(PADI4)mRNA的表达,分析其与RA患者的临床指标的相关性,探讨PADI4在RA发病机制中的作用及意义。方法实时定量PCR检测RA组(60例)、正常对照组(40例)PBMCs中PADI4 mRNA表达,并分析其与抗环瓜氨酸肽(CCP)抗体、疾病活动指数DAS28评分、C反应蛋白(CRP)、红细胞沉降率(ESR)、类风湿因子(RF)及病程等指标的关系。结果 RA组PADI4 mRNA相对表达[34.6(16.7,70.8)],明显高于正常对照组[20.6(11.1,51.8)](P<0.05)。RA组中PADl4 mRNA表达与抗CCP抗体、DAS28评分水平、ESR、RF呈正相关(r=0.527,P<0.001;r=0.416,P=0.001;r=0.371,P=0.004;r=0.287,P=0.030),与CRP、病程无相关性(r=0.015,P=0.919;r=0.064,P=0.625)。结论 RA病人外周血PBMCs PADI4 mRNA表达显著增高,并与抗CCP抗体水平、DAS28评分、ESR、RF呈正相关,可能是RA病情活动的一个有用的指标,PADI4可能在RA的发病和病理过程中起重要作用。     

RF、AKA和抗CCP抗体联检对类风湿关节炎诊断的临床价值

目的:探讨类风湿因子(Rheum atoid factor,RF)、抗角质蛋白抗体(antikeratin antibody,AKA)及抗环瓜氨酸肽(anti-cyc lic c itru llinated peptide,CCP)抗体对类风湿关节炎(rheum atoid arthritis,RA)的临床意义和早期应用价值。方法:对40例类风湿关节炎患者、30例系统性红斑狼疮和30名正常健康体检者进行RF、AKA、抗CCP抗体检测,应用速率散射比浊法测定RF,间接免疫荧光法检测AKA,ELISA法测定抗CCP抗体。结果:40例RA患者血清中,RF灵敏度和特异性分别为70.0%、90.0%,AKA灵敏度和特异性分别为35.0%、96.7%,抗CCP抗体灵敏度和特异性分别为85.0%、93.3%,联检RF、AKA及抗CCP抗体灵敏度和特异性分别为97.07%、99.8%。结论:RF、AKA和抗CCP抗体可作为诊断RA比较特异的血清学指标,三项指标联检可在一定程度上弥补RF对RA的诊断不足,提高RA的阳性诊断率,且有助于疾病的预后判断。     

类风湿因子及相关自身抗体对类风湿关节炎的诊断价值

目的探讨类风湿因子（RF）、抗环瓜氨酸肽抗体（抗CCP）及抗角蛋白抗体（AKA）联合检测对类风湿关节炎（RA）的临床诊断价值。方法对80例RA和65例非RA的其他自身免疫病患者检测RF、RF-IgM、抗CCP及AKA 4种指标。结果自身免疫性疾病女性发病较高,以RA组为甚;RA组的RF、RF-IgM、抗CCP及AKA高于其他自身免疫性疾病组;单指标检测RA诊断的敏感性为RF〉RF-IgM〉AKA〉抗CCP,特异性为RF-IgM〉抗CCP〉RF〉AKA。阳性预测值为抗CCP〉RF〉RF-IgM、AKA,阴性预测值为RF〉RF-IgM〉AKA〉抗CCP;以并联或串联方式联合检测均以RF、RF-IgM及抗CCP三联及RF、RF-IgM、抗CCP及AKA四联检测为佳,并联检测的敏感性分别为93%及97%,串联检测的特异性分别为98%及99%。结论 RF、RF-IgM、抗CCP及AKA 4种指标联合检测可提高对RA诊断的敏感性和特异性,对RA的早期诊断有较高价值。     

抗CCP抗体和AKA在类风湿关节炎诊断的应用价值

目的 比较抗环瓜氨酸肽抗体(抗CCP抗体)和抗角蛋白抗体(AKA)在类风湿关节炎诊断中作用,探讨RA的早期诊断方法.方法 对已确诊的85例RA患者、74例非RA的自身免疫病患者同时测定抗CCP抗体(ELISA法)、AKA(间接免疫荧光检测).结果 抗CCP抗体对诊断RA的灵敏度和特异性分别为75.3％和93.2％;AKA对RA诊断的灵敏度和特异性分别为89.4％和85.14％.抗CCP抗体的灵敏度与AKA灵敏度差异无统计学意义(P＞0.05),特异性差异有统计学意义(P＜0.05);抗CCP抗体或AKA与二者联合检测的灵敏度差异有统计学意义(P＜0.05),特异性差异有统计学意义(P＜0.05).阳性预测值抗CCP抗体较高,阴性预测值以二者联合检测较好,Youden index二者联合检测比单独检测抗CCP抗体或AKA高.抗CCP抗体和AKA在检测RA组中抗CCP抗体和AKA同时阳性检出58例;抗CCP抗体或AKA阳性共检出85例.抗CCP抗体或AKA阳性率(96.5％)比二者同时阳性率(68.2％)大大提高.结论 抗CCP抗体、AKA对RA具有较好的灵敏度和高度的特异性,联合检测抗CCP抗体和AKA可作为早期RA患者及RF阴性RA患者的早期诊断指标.     

抗环瓜氨酸肽抗体在类风湿性关节炎诊断中的应用

目的　探讨抗环瓜氨酸肽 (CCP)抗体检测在类风湿性关节炎 (RA)诊断中的意义。方法　根据cDNA序列人工合成的CCP为抗原 ,用ELISA方法检测 2 18例 (RA 112例 ,非RA 76例及正常人 30例 )血清中的抗CCP抗体 ,并比较抗CCP抗体与类风湿因子 (RF)的相关性。结果　抗CCP抗体在RA组患者血清中的阳性率 4 9.1% ,明显高于非RA组 ( 3.94 % )和正常对照组 ( 0 .0 % ) ,经 χ2 检验P <0 .0 5。抗CCP抗体对诊断RA的敏感性和特异性分别为 4 9.1%、96 .1% ,阳性预测值和阴性预测值为 94 .8%、4 3.8%。在 112例确诊为RA患者的血清中抗CCP抗体与RF重叠阳性率为 85 .4 % ,两者含量呈正相关。结论　抗CCP抗体对RA具有很高的特异性 ,可视为新的RA血清学诊断指标     

类风湿关节炎四种自身抗体的检测及意义

为评估类风湿因子(rheumatoid factor,RF)、抗环瓜氨酸肽(cyclic citrullinated pepdide,CCP)抗体、抗Sa抗体和抗角蛋白抗体(anti-keratin antibody,AKA)自身抗体对类风湿关节炎(rheumatoid arthritis,RA)诊断的意义,采用速率散...     

第二代抗环瓜氨酸肽抗体的检测在类风湿关节炎诊断中的意义

目的探讨抗环瓜氮酸肽（CCP）抗体的检测在类风湿关节炎（RA）诊断中的价值。方法ELISA法分别检测108例RA、89例非RA（其它风湿病患者）和78例健康体检者的抗CCP抗体；用间接免疫荧光法和速率散射比浊法检测抗角蛋白抗体（AKA）和类风湿因子（RF），分析CCP抗体的水平及与AKA、RF的相关性。结果抗CCP抗体的阳性率在RA中为87．4％（94／108），在非RA中为8．99％（8／89），正常人为0％（0／78）。3种抗体对RA诊断的敏感性和特异性分别为CCP87．4％、91．01％，AKA58．33％，82．24％，RF81．36％、75．35％。CCP抗体与AKA在RA患者血清中的阳性率之间差异非常显著，与RF差异不显著。3种方法的检测结果间存在相关性。结论用ELISA法检测血清中CCP抗体简便、结果可靠，对RA诊断具有高度的敏感性和特异性。     

类风湿关节炎六种指标联合检测的临床意义

目的评价分析抗瓜氨酸化波形蛋白(MCV)抗体、抗葡萄糖-6-磷酸异构酶(GPI)抗体等六项指标在类风湿关节炎(RA)患者血清中的表达与疾病的相关性及应用价值。方法收集110例RA、26例SLE、23例强直性脊柱炎(OA)、20例干燥综合征(SS)患者和50名健康人血清标本,用ELISA检测血清中抗MCV、抗GPI和抗环瓜氨酸肽(CCP)抗体水平,采用间接免疫荧光法检测抗角蛋白(AKA)抗体,采用散射比浊法检测类风湿因子(RF)和hs-CRP水平。结果 4种抗体在RA患者组中的阳性率显著高于非RA疾病组和健康对照组。除AKA抗体阳性率较低(41.8%)外,其余抗体在RA组中的阳性率差异无统计学意义。结论抗MCV和抗GPI抗体诊断RA的敏感度最高;抗CCP抗体的特异性最高,AKA抗体具有较高的特异性,但敏感度最低,联合检测可弥补单项抗体检测造成的漏诊。     

轻链λ对RF和抗CCP抗体阴性的类风湿关节炎的临床价值研究

为了评价类风湿因子(RF)和抗CCP抗体阴性的关节炎患者就诊时血清轻链λ水平对类风湿关节炎(RA)的诊断价值,回顾性分析我院住院RA患者的临床资料,获取患者初诊的RF、抗环瓜氨酸肽(CCP)抗体、免疫球蛋白(Ig)和轻链等数据。根据RF和抗CCP抗体水平,RA患者分为自身抗体阴性组和阳性组。将血清阴性RA患者作为实验组,与非RA关节炎患者构成的对照组进行比较,用受试者工作特征曲线(ROC)和多元Logistic回归分析轻链λ对RA患者诊断的临床价值。结果显示,实验组较对照组关节炎患者的轻链λ水平升高,差异具有统计学意义(P=0.026);ROC分析表明轻链λ对关节炎患者发展为RA具有预测价值,曲线下面积(AUC)达到0.665[95%CI(0.518,0.812)];以620.5mg/dl为临界值,预测RA的敏感性为0.565(0.345,0.768),特异性为0.804(0.661,0.906);多元Logistic回归校正后轻链λ仍然与关节炎患者发生RA相关(OR=3.551,P=0.037)。研究结果表明,血清轻链λ水平对关节炎患者发展为RA具有一定的预测价值。     

血清抗环瓜氨酸肽抗体与类风湿关节炎合并肺间质纤维化的相关性分析

分析类风湿关节炎(rheumatoid arthritis,RA)患者血清中抗环瓜氨酸肽(CCP)抗体水平与类风湿关节炎患者合并肺间质纤维化的关系.选取RA患者87例,其中合并肺间质纤维化(IPF)22例,单纯RA45例,采用酶联免疫吸附实验(ELISA)检测血清抗CCP抗体滴度,免疫比浊法检测类风湿因子(RF)滴度,...     

Prevalence and significance of antibodies to citrullinated human papilloma virus-47 E2345-362 in rheumatoid arthritis

It has been confirmed that antibodies to citrullinated profilaggrin(306-324) may play important roles in RA. In this study, human papilloma virus (HPV)-47 E2(345-362), homologous to profilaggrin(306-324), was found using the NCBI BLAST program. Then, E2(345-362) and citrullinated E2(345-362), with arginine(348) replaced by citrulline, were synthesized. The presence of antibodies against these peptides was examined by an enzyme-linked immunosorbent assay. Associations between these antibodies and the clinical and laboratory features of RA were evaluated. Although the prevalence and AU value of antibodies to the E2(345-362) peptide were similar in RA and other rheumatic diseases, those of antibodies to the citrullinated E2(345-362) peptide were significantly higher in RA than in other rheumatic diseases. Additionally, sera that were preincubated with cyclic citrullinated peptide (CCP) demonstrated lower AU values of anti-citrullinated E2(345-362) peptide antibodies. Moreover, the prevalence of anti-CCP antibodies and that of anti-peptidylarginine deiminase (PADI4) antibodies in anti-citrullinated E2(345-362)-positive patients were all higher than those of anti-citrullinated E2(345-362)-negative patients. There were significant correlations between anti-citrullinated E2(345-362) and anti-PADI4. RA patients with antibodies to citrullinated E2(345-362) had higher DAS28 scores, erythrocyte sedimentation rates, and radiographic progression than those without the antibodies. These results suggest that HPV-47 E2 may act as an autoantigen in RA. The increase in PADI4 may make it easier to citrullinate the HPV-47 E2(345-362) peptide, leading to the subsequent immune responses.     

Detection of antibodies against streptococcal peptidoglycan and the peptide subunit (synthetic tetra-D-alanyl-bovine serum albumin complex) in rheumatic-diseases.

Serum antibodies reactive with streptococcal cell wall peptidoglycan (PG) and its peptide subunit (synthetic tetra-D-alanine) were measured by enzyme-linked immunosorbent assay (ELISA) in patients with rheumatoid arthritis (RA), juvenile rheumatoid arthritis (JRA), osteoarthritis and acute rheumatic fever (RF) compared with healthy subjects. Using 'checkerboard' titrations, anti-PG antibody in human serum was detected at a concentration of PG antigen at 10 micrograms per well with serum dilutions of 1:1,000. For measurement of anti-tetra-D-alanine antibody, the antigen, (D-Ala4)31 was used at 0.5 micrograms per well and sera were diluted to 1:200. When the IgG antibody levels to the PG and the tetra-D-alanine of the sera of patients with RA, JRA and RF were compared with sera from healthy subjects, the sera of the patients had significantly higher levels than did healthy subjects. Antibody that reacted with the PG in serum was absorbed with purified group-specific C-carbohydrate (A-CHO), but A-CHO was not capable of absorbing anti-(D-Ala4)31 antibodies. Therefore, the peptide subunit should be used as antigen in order to measure the specific antibody to PG. Both anti-PG and anti-tetra-D-alanine antibody in human sera primarily belonged to the IgG2 subclass.     

抗GPI抗体和抗CCP抗体对RA诊断价值的比较分析

目的:评价抗环瓜氨酸肽(CCP)抗体和抗葡萄糖-6-磷酸异构酶(GPI)抗体对类风湿关节炎(RA)的诊断价值.方法:用酶联免疫吸附试验(ELISA)分别测定RA患者42例、其他风湿病患者32例以及健康对照者30例血清中的抗CCP抗体和抗GPI抗体,并应用R0C曲线比较两者对RA的诊断价值.结果:RA组血清抗CCP抗体水...     

抗环瓜氨酸肽抗体检测对类风湿关节炎的临床诊断价值

目的探讨抗环瓜氨酸肽抗体（抗CCP抗体）检测对类风湿关节炎（RA）诊断的意义。方法采用酶联免疫吸附试验（ELISA）检测115份人血清的抗CCP抗体,同时采用免疫透射比浊法定量检测类风湿因子（RF）,包括40例RA患者,45例其它风湿病患者,30名正常人;并分析抗CCP抗体与RF实验结果之间的相关性。结果在40例RA病人中,抗CCP抗体的阳性率为80.0%,在其它风湿病人中的阳性率为7.0%,抗CCP抗体对RA的敏感性和特异性分别为80.0%、96.0%,其敏感性高于RF,但差异无统计学意义（P〉0.05）,特异性明显高于RF（P〈0.05）。联合应用抗CCP抗体与RF进行诊断,二者均阳性时敏感性为65.0%,特异性为97.3%。抗CCP抗体与RF实验结果之间无相关性。结论抗CCP抗体对RA具有较好的敏感性和很高的特异性,可与RF相互补充,联合检测可提高对RA早期诊断的准确性。     

PADI4基因在肿瘤中表达的初步研究

目的本研究旨在通过检测PADI4在肿瘤中的表达特征和表达机制,初步探讨PADI4是如何参与肿瘤的发病过程的。方法 1.应用ELISA的方法检测各种恶性肿瘤患者血液中PADI4的表达水平。2.应用ELISA双抗体夹心方法检测各种恶性肿瘤患者血液中瓜氨酸化抗凝血酶的表达水平。3.应用RT-PCR的方法检测PADI4mRNA在卵巢癌组织中的表达。4.应用Taqman实时荧光定量RT-PCR的方法检测PADI4mRNA在卵巢癌组的表达量。5.应用免疫组化的方法检测PADI4蛋白在卵巢癌组织中的表达。结果 1.PADI4在乳腺癌、肝细胞癌、肺癌、食管癌、胃癌、结直肠癌、胰腺癌、卵巢癌、肾细胞癌子宫癌、前列腺癌和膀胱癌各种恶性肿瘤患者血清中与对照组相比有很高的表达,差异有统计学意义（P〈0.05）。而在甲状腺癌中PADI4表达比健康血清低。在乳腺癌、肝细胞癌、肺癌、胃癌、结直肠癌、胰腺癌、卵巢癌和肾细胞癌等手术后患者血中PADI4表达明显下降。2.cAT在乳腺癌、肝细胞癌、肺癌、食管癌、胃癌、结直肠癌、胰腺癌、卵巢癌、膀胱癌、甲状腺癌和前列腺癌大多数恶性肿瘤中表达很高（P〈0.05）。cAT在恶性肿瘤患者手术后表达下降。3.PADI4和cAT在肿瘤患者血液中表达的相关性通过χ2检验,在肝癌、肺癌、卵巢癌等肿瘤中PADI4表达和cAT的表达密切相关（P〈0.01）。4.PADI4mRNA在卵巢癌组织中有表达,良性肿瘤病变组织中未检测到PADI4 mRNA的表达。5.PADI4mRNA在卵巢癌组织中的表达量为1.57×102copies/ml,良性肿瘤病变组织中未检测到PADI4mRNA的表达。6.PADI4蛋白在卵巢癌组织中显著表达,良性肿瘤病变组织中无表达或者微弱表达。结论 1.通过大量临床标本的检测,我们发现在肝癌、肺癌、卵巢癌、子宫癌中PADI4和瓜氨酸化抗凝血酶大量表达,并且二者表达密切相关,表明PAID4可能通过对抗凝血酶瓜氨酸化而参与了某些肿瘤的发病过程。2.通过RT-PCR和Taqman实时荧光定量RT-PCR方法检测了卵巢癌组织和卵巢良性肿瘤组织中PADI4mRNA的表达,发现在卵巢癌组织PADI4mRNA是高表达的,揭示了PADI4和卵巢癌之间的相关性。3.通过免疫组化方法检测了卵巢癌组织和卵巢良性肿瘤组织中PADI4蛋白的表达,发现在卵巢癌癌细胞中PADI4蛋白是显著表达的,表明PADI4在卵巢癌发病过程中发挥了重要作用。4.卵巢癌组织中PADI4在基因转录水平和蛋白翻译水平表达都是升高的,表明PADI4基因转录水平的异常可能导致其蛋白质表达异常,从而启动了了卵巢癌的发病过程。     

Soluble CD4 in patients with rheumatoid arthritis and osteoarthritis

An ELISA was used to measure the soluble form of the leukocyte surface antigen CD4 (sCD4) in the sera and synovial fluids (SF) of patients with rheumatic diseases. Patients with rheumatoid arthritis (RA) had raised levels of sCD4 in both their sera and synovial fluid compared to age-matched healthy controls. In patients with osteoarthritis levels of sCD4 in SF and sera were lower than in RA but higher than in sera of healthy individuals. Mononuclear cells from the synovial fluid of RA patients were found to produce spontaneously high levels of sCD4, but autologous blood cells only produced comparable levels after in vitro stimulation with mitogenic lectin. In individual RA patients with active disease, serial sCD4 levels fell preceding clinical improvement. In three patients where serum sCD4 levels fell and clinical improvement occurred, subsequent small increases in serum sCD4 preceded increased clinical disease activity by up to 5 days. Synovial fluid levels of sCD4 correlated positively with soluble interleukin 2 receptor levels but no correlation was found with sCD8 levels. We conclude that the release of sCD4 reflects the involvement of T helper cells and macrophages in the pathogenesis of joint inflammation, especially in RA.     

A functional haplotype and expression of the PADI4 gene associated with increased rheumatoid arthritis susceptibility in Chinese

To evaluate the association of single-nucleotide polymorphisms (SNPs) in PADI4 mRNA with rheumatoid arthritis (RA) in a Chinese population, we examined the distribution of four exonic SNPs of the PADI4 gene (padi4_89*G/A, padi4_90*T/C, padi4_92*G/C and padi4_104*T/C) and PADI4 gene expression in 70 RA patients and 81 controls. Increased RA susceptibility was associated with the minor alleles of padi4_89 (P = 0.012), padi4_90 (P = 0.002), padi4_104 (P = 0.001) and the functional haplotype carrying the four minor alleles (P = 0.008). Human leukocyte antigen (HLA)-DRB1 shared epitope (SE) alleles were also associated with increased RA susceptibility, and the individuals with minor alleles of four exonic SNPs and SE alleles showed more increased RA susceptibility. The PADI4 expression was significantly higher in RA patients than in controls (P < 0.001). HLA-DRB1 SE alleles and the genotypes carrying the minor alleles of four SNPs were associated with increased PADI4 expression. It is concluded that PADI4 SNPs, functional haplotype and PADI4 expression may contribute to an inherited predisposition to RA in a Chinese population.     

联合检测HLA-DR4、抗CCP抗体和抗Sa抗体在类风湿关节炎诊断中应用价值

目的 探讨联合检测HLA-DR4、抗CCP抗体和抗Sa抗体在类风湿关节炎诊断中应用价值.方法 在2014年10月至2017年1月期间,选取我院就诊的109例RA患者作为研究对象,选取同期我院收治的94例非RA患者和90例健康体检者作为对照组.比较三组患者血清HLA-DR4、抗CCP抗体和抗Sa抗体的阳性率、三项指标联合检测对RA诊断学指标.结果 RA组患者血清HLA-DR4、抗CCP抗体和抗Sa抗体阳性率显著高于非RA组和对照组(P<0.05).在HLA-DR4、抗CCP抗体和抗Sa抗体三个指标中,抗CCP抗体的灵敏度和特异性最高(P<0.05).在联合检测中,HLA-DR4、抗CCP抗体和抗Sa抗体联合检测的灵敏度和特异性最高(P<0.05).结论 HLA-DR4、抗CCP抗体和抗Sa抗体联合检测可以提高对类风湿关节炎的诊断准确率,可作为临床辅助诊断手段.     

Serum-soluble selectin levels in patients with rheumatoid arthritis and systemic sclerosis

Soluble forms of selectins may play a regulatory role in inflammatory responses that are key to the pathophysiology of rheumatic diseases such as rheumatoid arthritis (RA) and systemic sclerosis (SSc). The aim of this study was to examine whether the elevated serum-soluble (s) selectin levels are associated with RA or SSc. Serum sE-, sL- and sP-selectin levels were measured by sandwich enzyme-linked immunosorbent assay in 34 RA patients, 30 SSc patients and 16 healthy subjects. The levels of sE-selectin were significantly higher in RA and SSc patients than those in healthy subjects. The sL-selectin level was significantly lower in RA patients compared to healthy subjects. Serum sP-selectin levels were not significantly different among the study groups. The active RA patients had significantly higher serum sE- and sL-selectin levels compared to inactive RA patients. Also, some correlations were observed between the serum selectin levels and measures of disease activity such as erythrocyte sedimentation rate and C-reactive protein in RA patients. The higher levels of sE-selectin were found in SSc patients with pulmonary fibrosis, and there was also a negative correlation between diffusion capacity for carbon monoxide and serum sE-selectin. Serum levels of selectins may provide a useful additional marker for disease activity in RA patients and for disease severity in SSc patients.