类风湿关节炎患者红细胞C3b受体数目发迹及基因多态性分析

目的 分析类风湿关节炎（ＲＡ）患者Ｃ３ｂ受体（Ｃ３ｂＲ）表达数目及Ｃ３ｂＲ基因多态性分布规律，探讨Ｃ３ｂＲ数目改变的可能原因。方法 分别测定６３例ＲＡ患者及健康献血员６０人的红细胞Ｃ３ｂ受体花环率（Ｃ３ｂＲＲ）及免疫复合物花环率（Ｃ３ｂＩＣＲ）。用限制性片段长度多态（ＲＦＬＰ）分析及Ｓｏｕｔｈｅｒｎ ｂｌｏｔｔｉｎｇ杂交技术检测Ｃ３ｂＲ基因多态性。结果 ＲＡ患者红细胞Ｃ３ｂ－ＲＲ和Ｃ３ｂ－ＩＣＲ分     

类风湿关节炎患者红细胞C3b受体数目改变及基因多态性分析

目的　分析类风湿关节炎（ＲＡ） 患者Ｃ３ｂ受体（Ｃ３ｂＲ） 表达数目及Ｃ３ｂＲ 基因多态性分布规律,探讨Ｃ３ｂＲ 数目改变的可能原因。方法　分别测定６３ 例ＲＡ 患者及健康献血员６０ 人的红细胞Ｃ３ｂ 受体花环率（Ｃ３ｂＲＲ） 及免疫复合物花环率（Ｃ３ｂＩＣＲ） 。用限制性片段长度多态（ＲＦＬＰ） 分析及Ｓｏｕｔｈｅｒｎ ｂｌｏｔｔｉｎｇ 杂交技术检测Ｃ３ｂＲ基因多态性。结果　ＲＡ 患者红细胞Ｃ３ｂＲＲ 和Ｃ３ｂＩＣＲ 分别显著低于和高于正常对照组。Ｃ３ｂＲ基因的两个等位基因基因型和表现型频率在ＲＡ 组和对照组均无显著差异。结论　ＲＡ 患者Ｃ３ｂＲ数目下降可能与基因变异无关,可能是受体基因转录和翻译过程异常所致。设法增加红细胞Ｃ３ｂＲ 数目以加速免疫复合物的清除,可能是治疗ＲＡ的有效途径之一。     

聚合酶链式反应与DNA探针检测临床标本中结核分支杆菌的评价

本文应用ＰＣＲ联合ＤＮＡ探针快速检测临床标本中结核杆菌ＤＮＡ。对临床３０７例标本同时进行ＰＣＲ和Ｓｏｕｔｈｅｒｎ杂交。     

结核性腹膜炎的实验室诊断

目的评价聚合酶链反应（ＰＣＲ）结合Ｓｏｕｔｈｅｒｎ杂交技术及酶联免疫吸附试验（ＥＬＩＳＡ）对结核性腹膜炎的诊断价值。方法用ＰＣＲ结合地高辛标记核酸探针Ｓｏｕｔｈｅｒｎ杂交技术检测４２例结核性腹水中结核分支杆菌ＤＮＡ，并与常规细菌学检测及ＥＬＩＳＡ对比。引物来自结核分支杆菌特异重复插入序列ＩＳ６１１０。特异性通过杂交及限制性内切酶ＳａｌⅠ酶切证实。同时比较了Ｓｏｕｔｈｅｒｎ杂交检测与凝胶电泳检测的敏感性。结果ＰＣＲ的敏感性为６９％，ＥＬＩＳＡ为７１％，培养为９％，涂片镜检均为阴性。并发现杂交较凝胶电泳检测更敏感。结论ＰＣＲ和ＥＬＩＳＡ法对结核性腹膜炎有较高的诊断价值，但前者更具有特异性。将Ｓｏｕｔｈｅｒｎ杂交技术与ＰＣＲ技术结合，可进一步提高检测的敏感性和特异性。     

类风湿性关节炎——血清IgA类风湿因子水平与类风湿性关节炎严重程度的相关性

重症类风湿性关节炎(ＲＡ)可伴有全身性损害而危及生命,目前有关ＲＡ关节外表现的本质研究尚少,对ＩｇＡ类风湿因子(ＲＦ)在ＲＡ关节外表现中的意义仍存在争议。本研究测定了ＲＡ病人的ＩｇＡＲＦ,评价它与ＲＡ临床特征的相关性。病人和方法　本研究的114例活动期ＲＡ均达到美国风湿病学院的修订标准。其中女91例、男23例,平均年龄519±117(18～76)岁,病程122±91(2～40)年,起病年龄395±121岁。29(254%)例有全身性表现,78例(684%)血清ＩｇＭＲＦ阳性。观察指标共56项,对ＩｇＡＲＦ阳性与阴性患者的临床特点进行对比分析。ＩｇＡＲＦ的定量…     

SLE患者血清中白细胞介素-2受体和睾酮水平的变化

选择５９例系统性红斑狼疮（ＳＬＥ）患者和２０例正常人,检测白细胞介素－２受体（ｓＩＬ－２Ｒ）和睾酮（Ｔｃ）水平。用疾病活动评分（ＳＬＡＭ）判断疾病活动性,并对ｓＩＬ－２Ｒ和Ｔｃ水平的变化及两者回的相关性进行分析。结果ＳＬＥ患者血清ｓＩＬ－２Ｒ水平显著高于正常人（Ｐ〈０．００１）,Ｔｅ显著低于正常人（Ｐ〈０．００１）,ＳＬＥ活动期ｓＩＬ－２Ｒ水平与ＳＬＡＭ指数显著高于非活动期（Ｐ〈０．０１）,Ｔｅ显     

P-选择素与类风湿关节炎及血小板升高

类风湿关节炎 (ＲＡ)是一种自身免疫性疾病。在病程中常可观察到血小板计数的升高 ,并与疾病的活动性相关。Ｐ 选择素是血小板活化的标志之一。我们通过检测ＲＡ患者外周血Ｐ 选择素水平 ,探讨Ｐ 选择素与血小板计数及ＲＡ疾病活动性的关系。一、资料和方法1 临床资料 :4 4例患者均符合 1987年美国风湿病协会修订的ＲＡ分类标准。男性 10例 ,女性 34例 ,平均年龄(48± 17)岁 ,病程 (36± 31)个月。其中 2 3例患者为ＲＡ活动期。ＲＡ活动期的标准为 :(1)≥ 3个关节肿胀 ;(2 )≥ 6个关节压痛 ;(3)晨僵≥ 30ｍｉｎ ;(4)血沉 (ＥＳＲ)≥ 2 8ｍｍ…     

类风湿关节炎患者T细胞受体BV基因的表达

目的 检测中国人群中类风湿关节炎（ＲＡ）患者Ｔ细胞ＴＣＲ ＢＶ基因表达的偏移。方法 采用半定量ＲＴ－ＰＣＲ方法,分析６例ＲＡ患者的外周血、关节液、滑膜（每例均取三种标本）中新鲜分离的未经ＩＬ－２刺激的Ｔ细胞ＴＣＲ ＢＶ１～２０基因片段的表达。结果 患者外周血与正常人外周血ＴＣＲ ＢＶ基因表达格局相似;患者关节液与滑膜ＴＣＲ ＢＶ基因表达格局相似;而患者关节内（关节液＋滑膜）ＴＣＲ ＢＶ基因表达格局     

强直性脊柱炎患者抗组蛋白抗体测定及其临床意义

以酶联免疫吸附试验及免疫印染技术对７８例强直性脊柱炎患者的血清进行抗总组蛋白及组蛋白亚单位抗体测定，发现抗总组蛋白抗体水平在ＡＳ组显著高于正常对照组及痛风性关节炎组，在活动期ＡＳ组高于非活动期组，其阳性率达２８％，在有虹膜炎的ＡＳ组显著高于无虹膜炎组，抗Ｈ３抗体与虹膜炎有显著性关联，结果提示高水平ＡＨＡ和Ｈ３抗体ＡＳ病情活动及与ＡＳ并发虹膜炎有关。     

聚合酶链反应对肠结核和克隆病的诊断价值

肠结核的诊断及其与克隆病的鉴别诊断颇为困难。在本研究中我们探讨了聚合酶链反应（ＰＣＲ）技术在这两方面的价值。３６例肠结核、２６例克隆病标本被纳入本研究。引物来自结核杆菌特异重复插入序列ＩＳ６１１０。其产物特导性通过Ｓａ１Ｉ限制性内切酶消化和地高辛标记的探针经Ｓｏｕｔｈｅｒｎ杂交证实。结果显示：在３６例肠结核中，２７例ＰＣＲ阳性１２６例克隆病中无１例阳性。从而证实了ＰＣＲ对肠结核是一种快速、敏感和特异的病原学诊断方法；并对肠结核与克隆病的鉴别诊断具有极高价值。     

Lytic Epstein-Barr virus infection in the synovial tissue of patients with rheumatoid arthritis

OBJECTIVE: To evaluate the existence of Epstein-Barr virus (EBV) infection in the synovial tissue of patients with rheumatoid arthritis (RA). METHODS: Synovial tissues were obtained at synovectomy or arthroplasty from 32 patients with RA and 30 control patients with osteoarthritis (OA). EBV DNA was detected by Southern blot hybridization and/or polymerase chain reaction (PCR) amplification. To localize the EBV-infected cells, tissue sections were studied by RNA in situ hybridization (ISH) for the EBV-encoded small RNA 1 (EBER-1), by DNA ISH for the Bam HI W region of EBV DNA, and by immunohistochemistry for EBV lytic proteins BZLF1 and gp350/220. RESULTS: EBV DNA was detected by PCR in 15 of the 32 samples from RA patients (47%), but in none of those from the 30 OA patients (P < 0.01). Of the 15 PCR-positive samples, 9 contained >1 EBV copy/1,000 cells (referred to as EBV 2+), and 6 contained 1 copy/1,000-5,000 cells (EBV 1+). Among the 9 EBV 2+ samples, 3 were also positive for EBV DNA by Southern blot hybridization, 5 were positive for EBER-1 by RNA ISH, and 3 were positive for EBV DNA by DNA ISH. Immunohistochemical analysis showed positive signals in all samples for BZLF1 and in 7 samples for gp350/ 220. In each examination, the positive signals were detected not only in lymphocytes, but also in synovial lining cells. CONCLUSION: EBV was frequently detected in the synovial tissue of RA patients. The infected cells were both lymphocytes and synovial cells, and expressed EBV proteins associated with virus replication. These findings suggest that EBV may play a role in the pathogenesis of RA.     

Citrullinated fibrinogen detected as a soluble citrullinated autoantigen in rheumatoid arthritis synovial fluids

BACKGROUND: Anti-citrullinated protein antibodies (ACPA) are specifically and frequently detected in sera of patients with rheumatoid arthritis (RA). Citrullinated fibrin or fibrinogen is a candidate autoantigen of such antibodies. OBJECTIVE: To investigate the presence of citrullinated fibrinogen (cFBG) in the plasma or synovial fluid of patients with RA and control patients, and to determine cFBG levels and their relationship with serum markers for RA if it is present. METHODS: A sandwich enzyme linked immunosorbent assay (ELISA) to measure cFBG was established using monoclonal antibodies cF16.1 and cF252.1, generated by immunising mice with R16Cit and R252Cit, the fibrinogen Aalpha chain derived sequences with citrulline at position 16 and 252, respectively, and the presence of cFBG was further investigated with immunoprecipitation-western blotting. RESULTS: Positive signals were detected in 11/15 RA synovial fluids (RASFs), but not in osteoarthritis synovial fluids or RA plasma with sandwich ELISA for cFBG using cF16.1 and an anti-modified citrulline (AMC) antibody. The presence of cFBG in RASFs was confirmed by immunoprecipitation-western blotting. Furthermore, most RA sera strongly reacted against R16Cit. No relationship was seen between RASF cFBG levels and C reactive protein or anti-cyclic citrullinated peptide antibody levels of the paired sera. CONCLUSION: cFBG is detected as a soluble citrullinated autoantigen in RASFs and may therefore be a genuine candidate antigen for ACPA in patients with RA.     

Ligase chain reaction in detection of Chlamydia DNA in synovial fluid cells

Synovial fluid cells from 12 patients with reactive arthritis (ReA) triggered by Chlamydia trachomatis were studied for the presence of Chlamydia DNA using the ligase chain reaction (LCR) LCx (Abbott) and the polymerase chain reaction (PCR) Amplicor (Roche). In addition, peripheral blood leucocytes from 11 of these patients were analysed by LCR. As controls, seven patients with newly diagnosed rheumatoid arthritis (RA) were included. Chlamydia trachomatis DNA was detectable by LCR in samples of synovial fluid cells from 4/12 patients with C. trachomatis-triggered ReA, and in none by PCR. Chlamydia trachomatis DNA was not detectable in the synovial fluid cells of the seven RA patients by either method, neither was C. trachomatis DNA detectable in the peripheral blood leucocytes of the ReA patients (0/11) or controls (0/6) by LCR. The LCR technique may be useful in the demonstration of Chlamydia DNA in synovial fluid cells.      

Bacterial DNA in synovial fluid cells of patients with juvenile onset spondyloarthropathies

OBJECTIVE: To identify bacterial DNA in synovial fluid cells of patients with active juvenile onset spondyloarthropathy (SpA). METHODS: The main group of study constituted 22 patients with juvenile onset SpA. In addition, five patients with adult onset SpA and nine with rheumatoid arthritis (RA) were studied. Polymerase chain reaction (PCR) with either genus- or species-specific primers was performed on synovial fluid cells to detect DNA sequences of Chlamydia trachomatis, Yersinia enterocolitica, Salmonella sp., Shigella sp., Campylobacter sp. and Mycobacterium tuberculosis. The presence of antibacterial antibodies in sera and synovial fluid was also determined by enzyme-linked immunoassay. RESULTS: The synovial fluid of nine patients with juvenile onset SpA, three with adult onset SpA and one with RA contained bacterial DNA. Five juvenile onset SpA samples had DNA of one single bacterium; two juvenile onset SpA and three adult onset SpA had DNA of two bacteria and two juvenile onset SpA had DNA of three bacteria. Overall, Salmonella sp. DNA was detected in seven synovial fluid samples, Shigella sp., Campylobacter sp. and M. tuberculosis were found in four samples each, and C. trachomatis was found in two. The bacterial DNA findings correlated with neither diagnosis nor disease duration. One RA synovial fluid had DNA of Campylobacter sp. Neither serum nor synovial fluid antibacterial antibodies correlated with DNA findings or clinical diagnosis. CONCLUSION: In this study, single and several combinations of bacterial DNA were identified in the synovial fluid of patients with long-term undifferentiated and definite juvenile onset SpA and adult onset SpA. Of relevance is that bacterial DNA corresponds to bacteria producing endemic disease in our population.     

Absence of Mycobacterium tuberculosis DNA in synovial fluid from patients with rheumatoid arthritis.

OBJECTIVES--To determine whether Mycobacterium tuberculosis DNA can be detected in synovial fluid of patients with rheumatoid arthritis (RA). METHODS--The polymerase chain reaction was applied to cellular components of synovial fluid. RESULTS--No evidence of M tuberculosis DNA was found in synovial fluid from 31 patients with RA and 13 control patients. CONCLUSION--The findings do not support a role for persistent M tuberculosis infection in the pathogenesis of RA.     

Cathepsin G: the significance in rheumatoid arthritis as a monocyte chemoattractant

Human cathepsin G (EC 3.4.21.20) has been reported to have the in vitro chemotactic activity for human monocytes. In this study, we examined the role of cathepsin G in monocyte involvement in joint inflammation of rheumatoid arthritis (RA) as a monocyte chemoattractant. Eighteen patients with RA and four patients with osteoarthritis (OA) were used in this study. Thiobenzylester substrate, Succ-Phe-Leu-Phe-S-Bzl, was used to measure the activity of cathepsin G in synovial fluids. Monocyte migration induced by cathepsin G and synovial fluids was assessed by a 48-well microchemotaxis chamber technique. Immunohistochemical staining was performed to determine the cellular origin of cathepsin G in RA synovial tissue. A very low activity of cathepsin G was detected in synovial fluids from patients with OA. On the other hand, significantly increased activity of cathepsin G was detected in patients with RA when compared with the value of OA patients. A considerable monocyte chemotactic activity was detected in the synovial fluid of RA patients, and the activity was partially decreased by the treatment with inhibitors for cathepsin G, α1-antichymotrypsin and phenylmethylsulfonyl fluoride. The activity of cathepsin G was significantly correlated with the neutrophil counts in synovial fluids and the concentration of interleukin-6. Immunohistochemical studies showed that cathepsin G was strongly expressed by synovial lining cells, and weakly expressed by macrophages and neutrophils in synovial tissues. This study indicates that the monocyte chemotactic activity of cathepsin G may have a role in the pathogenesis of RA synovial inflammation.     

Terminal complement complex in synovial tissue from patients affected by rheumatoid arthritis, osteoarthritis and acute joint trauma.

The C5b-9 complex (Terminal Complement Complex-TCC) is the final product of the terminal complement pathway. In this study, using the monoclonal antibody MCaE11 (specific for a C9 neoantigen) and an immunohistochemical technique, we examined the TCC deposits in synovial tissues from 4 patients affected by rheumatoid arthritis (RA) and 6 patients affected by osteoarthritis (OA). Synovial tissues from 8 patients affected by acute joint trauma were examined as controls. Furthermore, plasma TCC levels were measured in 44 RA patients and 51 controls, using the above mentioned antibody and a sandwich ELISA. Eight synovial fluids were also included in this study. Abundant TCC deposits were detected in the cytoplasm of the synovial lining cells and of large stromal mononuclear cells in all the RA and in 3 out of 6 OA synovial tissues characterized by histological signs of inflammation. No TCC deposits were found in non-inflamed synovial tissues from patients with joint trauma. In agreement with previous observations, the TCC plasma levels found were significantly higher in RA patients than in controls, but no difference was seen between patients with active and non-active disease. The mean TCC level was significantly higher in the synovial fluid than in the plasma, but no correlation emerged between these two series of values. This study shows that: a) the plasma level of TCCs cannot serve as an indicator of disease activity in RA; b) the TCC deposits in synovial tissue correlate well with the extent of inflammatory synovitis, irrespective of whether the synovitis is rheumatoid or osteoarthritic in nature.