原发性肝癌患者血清白细胞介素-12与外周血和癌组织中T淋巴细胞亚群变化的相关性研究

目的　探讨原发性肝癌患者血清白细胞介素 12 (IL 12 )与外周血和癌组织中T淋巴细胞亚群变化的规律及其相关性。方法　应用ELISA技术检测 36例原发性肝癌患者血清IL 12水平 ,采用流式细胞技术检测外周血T淋巴细胞亚群 ,免疫组化技术检测肝癌组织中T淋巴细胞亚群分布。结果　随着肝癌的发展 ,肿瘤体积增大和癌栓的形成 ,外周血CD 4 细胞数目减少 ,CD 8细胞数目增多 ,T4/T8比值进行性下降。血清IL 12水平和T4/T8比值间存在显著相关性。结论　IL 12是肝癌生物抗肿瘤的重要因素 ,T淋巴细胞亚群在抗肿瘤过程中有非常重要的作用     

恒河猴外周血调节性T淋巴细胞的分离纯化及鉴定

目的建立恒河猴外周血CD4+CD25+调节性T淋巴细胞(regulatory T cells,Tregs)快速有效的分离纯化方法。方法 4～5岁健康恒河猴10只,雌性4只,雄性6只,体重5～8 kg;于大隐静脉抽取外周血,每只抽取8 mL。密度梯度离心法分离外周血单个核细胞(peripheral blood mononuclear cell,PBMC),分别采用非人灵长类Tregs分离试剂盒的生物素标记的混合抗体和磁珠标记的抗生物素抗体阴性分选,以及大鼠抗人CD4-活化蛋白C(activatedprotein C,APC)和抗APC多选试剂盒中的磁珠标记的抗APC抗体阳性分选出CD4+T淋巴细胞,比较两种方法获得细胞的得率、活性及纯度;选择得率、活性和纯度较高的分选方法获得的CD4+T淋巴细胞,用磁珠标记的抗人CD25抗体进行阳性分选,获得CD4+CD25+Tregs,流式细胞仪检测纯度、活性及FoxP3表达水平,以及Tregs对刀豆蛋白(concanavalinA,ConA)刺激的自体CD4+CD25-效应性T淋巴细胞(effective T cells,Teffs)增殖的抑制功能。结果 CD4+T淋巴细胞阳性分选和阴性分选后,细胞活性均达95%左右,差异无统计学意义(P>0.05),但阳性分选后CD4+T淋巴细胞得率和纯度均显著高于阴性分选(P<0.05)。后续CD4+CD25+Tregs分选采用阳性分选法获得的CD4+T淋巴细胞。经双阳性分选收集的目的细胞中CD4+CD25+Tregs占76.2%±8.6%,活细胞比例为93.3%±4.7%,FoxP3阳性细胞比例为74.2%±6.9%。混合培养后Tregs均对ConA刺激的Teffs的增殖有抑制作用。结论免疫磁珠双阳性分选法能有效分选出恒河猴外周血中有功能的CD4+CD25+Tregs。     

环瓜氨酸多肽对类风湿关节炎患者外周血淋巴细胞分泌INF-γ及IL-4的影响

目的观察环瓜氨酸多肽(CCP)对类风湿关节炎(RA)患者外周血T淋巴细胞分泌INF-γ、IL-4的影响。方法分离培养24例RA患者和16例健康人外周血淋巴细胞,分为以下3组处理,RA空白对照组:RA患者外周血淋巴细胞分离培养不加任何药物干预;RA CCP组:RA患者外周血淋巴细胞加CCP干预(CCP终浓度为20μg/ml);健康空白对照组:健康人外周血淋巴细胞分离培养不加任何药物干预。上述各组细胞培养72 h后,酶联免疫吸附法(ELISA)检测细胞培养上清中IFN-γ及IL-4水平。结果与健康空白对照组外周血T淋巴细胞分泌的IFN-γ、IL-4及IFN-γ/IL-4相比,RA空白对照组IFN-γ水平升高(P0.05),IL-4水平下降(P0.01),IFN-γ/IL-4升高(P0.01);与RA空白对照组相比,RA CCP组IFN-γ水平进一步升高(P0.05),IL-4水平进一步下降(P0.05),IFN-γ/IL-4进一步升高(P0.01)。结论 CCP对RA患者外周血T淋巴细胞分泌Th1及Th2型细胞因子IFN-γ及IL-4有明显的调节作用,可能在RA发病机制中起重要的作用。     

CTGF刺激人增生性瘢痕成纤维细胞STATs蛋白活化的状况

目的：探讨CTGF刺激人增生性瘢痕成纤维细胞STATs蛋白活化的情况。方法：原代培养人增生性瘢痕成纤维细胞，将其分为：①对照组：人增生性瘢痕成纤维细胞纽；②增生性瘢痕成纤维细胞外源性重组CTGF刺激组。取0min、5min、10min、20min、30min、45min、60min、90min八个相点，时分别采用MTT检测细胞增殖，western-blot检测各组之间a-平滑肌肌动蛋白（a-smoothmuscle actin，a-SMA）的变化趋势以及各时相点STATl、STAT2、STAT3、STAT4、STAT5、STAT6蛋白活化（磷酸化）情况。结果：在确定成纤维细胞增殖以及a-SMA表达变化趋势以②-①组顺序递减的前提下（P〈0．05），用western—blot观测两组STATs蛋白活化情况，发现：STATl蛋白磷酸化水平以②-①组顺序递减（P〈0．01）；STAT2-6均有不同程度的磷酸化，但不以②-①组顺序递减。结论：STAT1可能在CTGF促进人增生性瘢痕成纤维细胞增殖和分化过程中可能发挥着重要作用，以上研究结果为抑制瘢痕纤维化及挛缩提供了新的研究方向。     

显性调控相关早反应基因的大规模扫描和分析

目的：动态观察白细胞介素10（IL－10）作用后，淋巴细胞早反应基因表达谱变化，为下一步评价患者免疫状态和诱导治疗提供理论依据。方法：应用DNA微矩阵技术，分离正常人外周血单个核细胞（PBMC），予以抗CD3McAb及CD3McAb IL-10刺激，抽提24h PBMC mRNA，反转录后与芯片杂交，通过生物信息学方法比较分析，结果：培养24h后，抗CD3McAb IL-10组与抗CD3McAb组比较有15个基因发生改变，5个基因表达明显下调，10个基因上调，主要功能在于促进基因编码区连接，影响 细胞骨架和运动，不同部位的磷酸化，去磷酸化，细胞识别，会促进细胞生长作用，细胞凋亡，信号传递的基因则重点涉及钙离子－钙调蛋白，G－蛋白信号通路。结论：在IL－10作用，细胞凋亡，信号传递的基因则重点涉及钙离子－钙调蛋白，G－蛋白信号通路。结论：在IL－10作用下，一系列基因转录活化，其中许多先前与免疫细胞无明确功能的相关基因参与了T细胞活化与调控，是否该群基因即代表IL－10作用下淋巴细胞的基因表达标签，尚须进一步证实；要完全明确T细胞调控网络，还应全面分析T淋巴细胞基因表达谱，从中筛选差异有显著意义的基因，进行功能研究。     

活化的挤血T淋巴细胞内细胞因子特性研究

目的;通过对人脐血中CD4^ 和CD8^ T淋巴细胞经特异抗原刺激后,胞内细胞因子γ－IFN和IL－4分泌水平的研究,探讨人脐血干细胞移植后急、慢性移植物抗宿主病（GVHD）发生低下的可能机制。方法;15份脐血和15份健康成人外周血中T淋巴细胞在莫能霉素存在的情况下,体外经十四烷酰指波酯乙脂和离子霉素刺激后,分别进行CD4－FITC、CD8－FITC荧光单抗染色和γ－IFN－PE、IL－4－PE荧光单抗胞内染色,最后进行流式细胞仪分析。结果：脐血中CD4^ Th细胞和CD8^ Tc细胞内的γ－IFN分泌水平均低于成人外周血中CD4^ 和CD8^ T细胞,而脐血CD8^ T细胞胞内IL－4分泌水平与成人外周同类细胞差异无显著性。结论：人脐血中T淋巴细胞受特异抗原刺激后,胞内不能产生正常的Th1/Tc1 样细胞因子谱,即活化的脐血T淋巴细胞Th1/Tc1样反应低下,这可能是脐血移植后GVHD发生低下的原因之一。     

胃癌患者外周血调节性T细胞的变化及其意义

目的 探讨胃癌患者外周血中CD4+CD25+T淋巴细胞、CD8+CD28-T淋巴细胞比例的变化及其临床意义.方法 采用流式细胞技术检测30例胃癌患者外周血中CD4+CD25+T淋巴细胞、CD8+CD28-T淋巴细胞玎分比,对照组为30例慢件胃炎患者.结果 胃癌患者外周血中CD4+CD25+调节性T细胞占T淋巴细胞的百分比为8.7%±1.3%,与胃炎患者相比差异无统计学意义.胃癌患者外周血中CD8+CD28-调节性T细胞占T淋巴细胞的百分比为30.3%±3.3%,明显高于胃炎患者的20.3%±2.7%.外周血中CD4+CD25+T淋巴细胞、CD8+CD28-T淋巴细胞百分比与胃癌淋巴结转移、病理分型无明显相关性.结论 外周血中调节性T细胞在胃癌发展中可能发挥一定作用.     

信号传导与转录激活因子-4基因rs7574865位点单核苷酸多态性与类风湿关节炎易感性荟萃分析

目的：综合评价信息传导与转录激活因子-4基因（STAT4基因）rs7574865位点单核苷酸多态性与类风湿关节炎易感性的相关性。方法：计算机检索Pubmed、Embase、中国生物医学文献数据库和万方数据库数据库等,并手工检索相关杂志,收集有关不同人群STAT4基因rs7574865位点单核苷酸多态性的等位基因频率、基因型频率与类风湿性关节炎易感性相关性的研究,采用Stata 12.0软件进行Meta分析;选择的遗传模型包括GT＋TT与GG比较,TT与GT＋GG比较,TT与GG比较以及等位基因T与G之间比较。结果：STAT4基因rs7574865位点T等位基因频率与类风湿性关节炎易感性可能具有相关性（OR=1.259,95%CI=1.202-1.319, P＜0.001）;T等位基因频率与类风湿性关节炎易感性的相关性在欧洲、亚洲、非洲、拉丁美洲人群中有统计学意义;在总体人口中STAT4基因rs7574865位点的基因型与类风湿性关节炎易感性可能具有相关性;在总体人口中STAT4基因rs7574865位点多态性与抗CCP抗体阳性或RF因子阳性类风湿性关节炎易感性仍可能具有相关性。结论：STAT4基因rs7574865位点单核苷酸多态性与类风湿性关节炎易感性可能具有相关性,而且这种相关性可能独立于患者RF因子以及抗CCP抗体水平。     

淋巴细胞特异蛋白质酪氨酸激酶/P38信号传递途径在小鼠肾小管上皮细胞的作用

目的 探讨小鼠肾上管上皮细胞是否存在淋巴细胞特异蛋白质酪氨酸激酶（lck）基因表达及lck/p38细胞分裂原活化蛋白信号传递分子经IL-12、IL-2、脂多糖（LPS）等刺激时在肾小管上皮细胞的变化。方法 无菌分离培养BALB/C小鼠肾小管上皮细胞,采用地高辛标记的lck寡核苷酸探针原位杂交检测肾小管上皮细胞中lck基因表达;应用放射自显影方法观察lck活性变化,利用免疫印迹分析lck蛋白表达及p38磷酸化。结果 IL-12、IL-2、LPS刺激肾小管上皮细胞活化,lck mRNA表达较对照组明显增强;经上述物质刺激后lck活性、p38磷酸化分别于5min、15min最强,其中LPS刺激最显著而lck、p38蛋白水平则无明显变化。加入lck抑制剂PP1,IL-12刺激时未发现p38磷酸化,而IL-2、LPS刺激只有轻度抑制。结论 IL-12、IL-2、LPS促进肾小管上皮细胞中lck基因表达,并只有IL-12唯一通过lck诱导p38磷酸化,它们皆可通过lck/p38信号传递途径参与对肾小皮细胞发挥生物作用。     

逆转录病毒携带白细胞介素-12转染树突状细胞体外诱导特异免疫杀伤肝癌细胞

目的探讨逆转录病毒携带白细胞介素(IL)12转染树突状细胞(DC)体外诱导免疫杀伤肝癌细胞的效能及其机制。方法感染指数(MOI)为100,含IL12的重组逆转录病毒修饰肝癌患者外周血DC(DCIL12),酶联免疫吸附试验法(ELISA)检测DCIL12(5×103个/ml)培养上清中IL12和γ干扰素(IFNγ)水平,以冻融肝癌细胞株HepG2(1×107个/ml)所获得的肿瘤相关抗原(TAA)刺激DCIL12,MTT法检测TAA负载的DCIL12刺激同源T淋巴细胞(1×105个/ml)增殖分化能力,细胞毒试验检测DCIL12诱导的细胞毒T淋巴细胞(CTL)及其上清液对HepG2肝癌细胞株杀伤作用,ELISA法检测CTL上清液IFNγ水平。结果DC经IL12基因修饰后48h分泌高水平IL12(24.35±5.40)ng/L及IFNγ(725±30)ng/L,均显著高于未转染DC组(P<0.01及P<0.05)。DCIL12诱导的CTL及其上清液对HepG2均有显著杀伤作用,杀伤率显著高于未转染DC组,分别为(82.43±8.70)%vs(57.4±4.3)%(P<0.01)和(76.45±8.50)%vs(18.23±5.30)%(P<0.01),DCIL12诱导的CTL上清液IFNγ水平显著高于未转染DC组,分别为(1125.0±32.7)ng/Lvs(281.3±14.7)ng/L(P<0.01)。结论IL12基因修饰增强DC体外诱导自体T淋巴细胞产生特异性抗肝癌免疫,其机制与IL12基因修饰促进DC分泌IL12、增强T淋巴细胞活化及分泌IFNγ密切相关。     

Distinct serum and synovial fluid interleukin (IL)-33 levels in rheumatoid arthritis, psoriatic arthritis and osteoarthritis

ObjectivesRecent evidence suggests a role for interleukin (IL)-33 and its receptor ST2 in arthritis. In this study, we quantified IL-33 and soluble (s)ST2 levels in serum and synovial fluid (SF), and assessed synovial IL-33 expression levels and pattern in patients with rheumatoid arthritis (RA), psoriatic arthritis (PsA), or osteoarthritis (OA).MethodsSerum and SF IL-33 and sST2 levels were assessed by ELISA. IL-33 mRNA was quantified by RT-qPCR. Synovial IL-33 protein expression pattern was examined by immunohistochemistry.ResultsSerum and SF IL-33 levels tended to be higher in RA than in OA patients. In contrast to RA, IL-33 was not detectable in PsA serum and SF. Serum sST2 levels were higher in RA than in OA. There was a wide variation of synovial tissue IL-33 mRNA expression within each disease group and IL-33 mRNA levels were not significantly different between the groups. A similar IL-33 protein expression pattern was observed in RA, PsA and OA synovium, with strong nuclear expression of IL-33 in endothelial cells and, in a subset of RA, PsA and OA patients, in cells morphologically consistent with synovial fibroblasts.Discussion/ConclusionsThis study confirms increased circulating IL-33 levels in RA. In addition, we report that IL-33 is undetectable in the serum or SF of PsA patients. Local expression of IL-33 in the synovium was observed at similar variable levels in RA, PsA and OA, suggesting that inflamed joints do not represent the primary source of elevated serum and SF levels of IL-33 in RA.     

Increased risk of revision for infection in rheumatoid arthritis patients with total hip replacements

Background and purpose — Medical treatment of rheumatoid arthritis (RA) has changed dramatically over the last 15 years, including immune modulation. We investigated the risk of revision for infection after primary total hip replacement (THR) in patients with rheumatoid arthritis over a 16-year period, and compared it with that in THR patients with osteoarthritis (OA).

Patients and methods — We identified 13,384 THRs in RA patients and 377,287 THRs in OA patients from 1995 through 2010 in a dataset from the Nordic Arthroplasty Register Association (NARA). Kaplan-Meier survival curves, with revision for infection as the endpoint, were constructed. Cox regression analyses were performed to calculate the relative risk (RR) of revision for infection adjusted for age, sex, fixation technique, and year of primary surgery.

Results — RA patients had a 1.3 times (95% CI 1.0–1.6) higher risk of revision for infection. After 2001, this risk increased more for RA patients than for OA patients. During the first 3 months and from 8 years postoperatively, the risk of revision for infection was higher in RA patients with THRs fixated with antibiotic-loaded cement than in corresponding OA patients.

Interpretation — We found a slightly higher overall risk of revision for infection in RA patients than in OA patients, but this difference was only present after 2001. In THRs with antibiotic-loaded cement, the risk of very early and late infections leading to revision was higher in RA patients than in OA patients.     

Difference between phenotypes of bone marrow and peripheral blood leukocytes indicate rheumatoid arthritis bone marrow as an important site for cell activation and proliferation

ObjectivesAlthough the pathogenesis of RA is still unknown, recent data suggest bone marrow compartment is an important site contributing to the initiation and perpetuation of chronic inflammation that may spread to the joint. The aim of the present study was to compare phenotypes of bone marrow and peripheral blood leukocytes isolated from rheumatoid arthritis (RA) and osteoarthritis (OA) patients that may indicate whether bone marrow is implicated in the activation and propagation of mature leukocytes.MethodsMononuclear cells were isolated from peripheral blood and bone marrow of patients with RA and OA undergoing hip replacement. Sodium citrate was used as an anticoagulant. To assess cell phenotypes, specific monoclonal antibodies against: CD3, CD19, CD20, CD4, CD8, CD16, CD56, CD38, CD27, CD25, CD69, CD14, CD123, CD11c, HLA-DR, BDCA-1 and BDCA-2 were used for staining, followed by flow cytometric analysis.ResultsPreliminary results indicate that RA bone marrow compartment contained different proportions of activated CD4+ and CD8+ T cell, B-cells, NK-cells and macrophages than paired peripheral blood. Unlike in peripheral blood from RA and OA, where relatively small differences in cell phenotypes were detected, composition of bone marrow mononuclear cells differed substantially between these two diseases.ConclusionsPhenotypic differences between leukocytes from bone marrow and peripheral blood indicate that in RA bone marrow microenvironment provides stronger signals leading to activation and maturation of leukocytes than in OA. Thus, these data support notion that bone marrow actively participates in the pathogenesis of RA.AcknowledgementsThis work was partially supported by funds from the European Community's FP6 Project 018661 Autocure.     

类风湿关节炎及骨关节炎血清和滑液中一氧化氮的测定

目的 研究一氧化氮（ＮＯ）在类风湿性关节炎（ＲＡ）及骨关节炎（ＯＡ）病变中的作用。方法 采用亚硝酸盐间接法测定了４０例ＲＡ、２０例ＯＡ患者表及滑液ＮＯ的含量变化,并与年龄相匹配的健康人相比较。结果 ＲＡ患者血清及滑液ＮＯ的浓度呈正ｒ＝０．４１７,Ｐ〈０．０５）。ＲＡ和ＯＡ患者血清ＮＯ浓度均低于滑液中的,但显著高行正常对照组血清ＮＯ浓度;ＲＡ患者滑液ＮＯ浓度高于血清中,且明显高于ＯＡ组滑液ＮＯ浓度。     

Increased sensitivity of rheumatoid synoviocytes to Schnurri-3 expression in TNF-α and IL-17A induced osteoblastic differentiation

ObjectiveTo compare the effects of TNF-α and IL-17A on osteogenic differentiation of isolated fibroblast-like synoviocytes (FLS) from healthy donors, osteoarthritis (OA) and rheumatoid arthritis (RA) patients.MethodsFLS were cultured in osteogenic medium, with and without TNF-α and/or IL-17A. Extracellular matrix mineralization was evaluated by alizarin red staining and alkaline phosphatase activity (ALP) measurement. mRNA expression was analyzed by qRT-PCR for Wnt5a, BMP2 and Runx2, genes associated with osteogenesis, for DKK1 and RANKL, genes associated with osteogenesis inhibition and Schnurri-3, a new critical gene in the cross talk with osteoclasts. IL-6 and IL-8 production was measured by ELISA.ResultsIn osteogenic medium, matrix mineralization and increased ALP activity indicated that FLS can undergo osteogenic differentiation, which was increased with TNF-α and IL-17A. The expression of osteogenesis activators (BMP2 and Wnt5a) was increased with cytokines and that of the osteogenesis inhibitor DKK1 was decreased. There was no difference between all three cell types. In contrast, RA FLS were particularly sensitive to the synergistic increase of Shn3 with TNF-α and IL-17A. Levels of IL-6 and IL-8 were also higher for RA-FLS, compared to healthy and OA FLS.ConclusionIL-17A and/or TNF-α treatment favor an osteogenesis induction in isolated FLS, independent of their origin. RA-FLS were more sensitive to the synergistic increase of Schnurri-3 expression. Combined with the higher levels of inflammation, this may in turn activate osteoclastogenesis, leading to increased bone destruction seen in destructive arthritis.     

Hand impairment and activity limitations in four chronic diseases

Study designRetrospective cohort.IntroductionHand involvement in osteoarthritis (OA) and rheumatoid arthritis (RA) are well known to occupational and physical therapists; however, it is not known whether the impairments and activity limitations with diabetes (DMII) and systemic sclerosis (SSc) are as severe as those observed with OA and RA.PurposeTo compare the hand impairments and activity limitations in the 4 diseases.MethodsA convenience sample of 156 participants received evaluations of hand impairments: strength, joint motion, and dexterity and completed a hand activity limitations questionnaire.ResultsThe SSc and RA participants had weaker pinch, decreased joint motion and more activity limitations than the DMII and OA groups. There were no significant differences between the groups for right hand grip strength and pegboard dexterity, and applied dexterity.ConclusionsOA and DMII groups had significantly less impairments and activity limitations than the SSc and RA groups.Level of evidence2C.     

The increased in vitro osteoclastogenesis in patients with rheumatoid arthritis is due to increased percentage of precursors and decreased apoptosis - the In Vitro Osteoclast Differentiation in Arthritis (IODA) study

Increases in local and systemic bone resorption are hallmarks of rheumatoid arthritis (RA). Osteoclasts are implicated in these processes and their enhanced differentiation may contribute to bone destruction. We observed that in vitro osteoclastogenesis varies among healthy individuals and hypothesized that increased osteoclastogenesis could be a marker for the presence of RA. Our objective in the present study was to determine if in vitro osteoclastogenesis from peripheral blood mononuclear cells (PBMCs) was different in patients with RA compared to healthy controls and osteoarthritis (OA) patients. Expression of CD14 in PBMCs was quantified and PBMCs were incubated for 21 days in the presence of the osteoclastogenic cytokines M-CSF and RANKL. Differentiation on cortical bone slices permitted the analysis of bone resorption while apoptotic potential was assessed by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling. In vitro osteoclastogenesis was higher in PBMCs from RA patients compared to controls, and a similar increase was observed in the percentage of osteoclast precursors in RA patients. Osteoclasts from RA patients showed lower apoptotic rates than osteoclasts from healthy controls. No difference was observed in bone resorption activity between RA patients and controls. Interestingly, the difference in osteoclast number and apoptosis rate allowed the implementation of an algorithm capable of distinguishing patients with RA from controls. In conclusion, our study shows that osteoclast differentiation from PBMCs is enhanced in patients with RA, and this difference can be explained by both a higher percentage of osteoclast precursors in the blood and by the reduced apoptotic potential of mature osteoclasts.     

Involvement of interleukin-6 and prostaglandin E2 in periarticular osteoporosis of postmenopausal women with rheumatoid arthritis

In experimental arthritis, blocking of the receptor activator of nuclear factor κB ligand (RANKL) by osteo-protegerin (OPG) treatment prevents bone loss but not inflammation, suggesting that there are inflammation-related factors that regulate RANKL and OPG. However, it is not known which factors regulate RANKL and OPG in human inflammation-induced bone loss. To clarify the inflammation-related factors that play a role in periarticular osteoporosis in patients with rheumatoid arthritis (RA), the synovial fluid and synovium of the knee joint, and the periarticular cancellous bone of the femoral condyle were collected at surgery from postmenopausal women with RA or osteoarthritis (OA). All patients with RA had radiologic bone loss on the femoral condyles, while such a loss was not observed in patients with OA. The present study examined: (i) tumor necrosis factor α (TNFα), interleukin (IL)-1β and IL-6 levels in synovial fluid; (ii) TNFα, IL-1β and IL-6 messenger RNA (mRNA) expression in the synovium and the cancellous bone that contained bone marrow; and (iii) IL-6 and prostaglandin E₂ (PGE₂) production in cultured osteoblast-lineage cells derived from collagenase-treated cancellous bone fragments. Inflammation of the knee joints in patients with RA was confirmed by significantly higher proinflammatory cytokine levels in the synovial fluid and the synovium than those seen in patients with OA. In patients with RA, mRNA expression of IL-6, but not TNFα and IL-1β, in the cancellous bone and IL-6 and PGE₂ production in the osteoblast-lineage cells were significantly higher than in patients with OA. These findings suggest, for the first time, that IL-6 is involved in periarticular osteoporosis in postmenopausal women with RA. IL-6 and PGE₂ released from osteoblast-lineage cells could be, at least in part, responsible for human inflammation-induced bone loss. Received: August 28, 2000 / Accepted: November 9, 2000     

Ultrasonographic criteria for the diagnosis of erosive rheumatoid arthritis using osteoarthritic patients as controls compared to validated radiographic criteria

ObjectivesThe aims of this study were to compare characteristics of radiography (RX) and ultrasound (US) erosive lesions in rheumatoid arthritis (RA) and osteoarthritis (OA) patients (prevalence, topography and severity), to determine thresholds for the diagnosis of erosive RA based on US and to evaluate the performance of US and RX to establish a diagnosis of erosive RA differentiated from hand OA.MethodsPatients fulfilling ACR 1987 and/or ACR/EULAR 2010 criteria for RA or ACR hand OA criteria were prospectively included. A modified Sharp erosion score was assessed by two blinded readers and one adjudicator for discordant cases (number of eroded joints ≤ three). Erosions in US were scored on six bilateral joints (MCP2-3, 5; MTP2-3, 5) with a four-grade scale to calculate total US score for erosions (USSe).ResultsA total of 168 patients were included: 122 RA (32 early RA < 2 years; 90 late RA ≥ 2 years); 46 OA patients. On RX: 42 RA patients (6 early; 36 late) and 5 OA patients were eroded according to EULAR 2013 definition criteria with sensitivity at 34.4% and specificity at 89.1%. On US, 95 RA patients (21 early; 74 late) and 12 OA patients were eroded. Considering at least two joint facets eroded or at least one joint facet eroded at grade 2 on US, sensitivities were good (68–72.1%) and specificities excellent (89.1–100%). Agreement between RX and US was excellent (90–92%). The positive and negative likehood ratios were respectively 3.16 and 0.73 for radiography and 6.64 and 0.31 for US (for two facets eroded).ConclusionUSSe can differentiate RA from OA in erosive disease and detect two times more patients with erosive RA than RX with excellent specificity and agreement.     

类风湿关节炎滑膜成纤维细胞通过高表达RANKL促进破骨细胞分化及活化

目的探讨类风湿关节炎滑膜成纤维细胞（RA-FLS）对破骨细胞（Oc）分化和活化的作用及机制。方法活动期RA滑膜体外分离培养FLS,以骨关节炎（OA）-FLS为对照,分别与健康人外周血单核细胞（MNC）共培养后,抗酒石酸酸性磷酸酶（TRAP）染色鉴定Oc并计数、甲苯胺蓝染色观察骨吸收陷窝情况。细胞免疫荧光染色检测FLS RANKL表达,Real-time PCR及W estern blot检测RANKL和OPG mRNA、蛋白表达。结果 RA-FLS与MNC共培养7 d时TRAP＋且细胞核≥3个的Oc很少,14 d时见较多的Oc,21 d甲苯胺蓝染色示清晰的骨吸收陷窝,而OA-FLS与MNC共培养后未见Oc,也未见骨吸收陷窝。细胞免疫荧光染色示RA-FLS较OA-FLS高表达RANKL（P〈0.05）。RA-FLS RANKL mRNA和蛋白表达较OA-FLS明显增高,而OPG mRNA和蛋白表达则明显降低,RANKL/OPG mRNA和蛋白比率较OA-FLS明显增高（均P〈0.05）。结论 RA-FLS可能通过高表达RANKL,促进外周血MNC向Oc分化,并促进Oc的骨吸收功能。