类风湿关节炎中转化生长因子β1和细胞间粘附分子的变化

目的：了解类风湿关节炎（ＲＡ）血清、滑液和滑膜组织中转化生长因β１（ＴＧＦβ１）、细胞间粘附分子１（ＩＣＡＭ１）及细胞间粘附分子３（ＩＣＡＭ３）的变化。方法：采用ＥＬＩＳＡ及ＡＢＣ免疫组化方法检测ＲＡ患者血清、滑液和滑膜中ＴＧＦβ１、ＩＣＡＭ１和ＩＣＡＭ３的浓度和阳性程度。结果：ＲＡ血清中ＴＧＦβ１含量较低，而ＩＣＡＭ１含量明显升高，ＩＣＡＭ３含量正常，滑液中ＩＣＡＭ３含量低于血清含量。ＲＡ血清中ＴＧＦβ１含量与ＩＣＡＭ１含量呈中度负相关，与ＩＣＡＭ３含量无显著相关。在ＲＡ滑膜中巨噬细胞、滑膜衬里细胞和成纤维细胞ＴＧＦβ１染色阳性，巨噬细胞、滑膜衬里细胞、成纤维细胞和血管内皮细胞ＩＣＡＭ１染色阳性。结论：ＴＧＦβ１和ＩＣＡＭ１参与了ＲＡ的发生和发展过程，在ＲＡ慢性炎症中，ＩＣＡＭ１对炎细胞转移并聚集于滑膜可能有重要作用，ＲＡ外周血ＴＧＦβ１浓度减低伴随ＩＣＡＭ１浓度的增加。     

转化生长因子-β1在肝细胞性肝癌中表达增强

目的转化生长因子－β１（ＴＧＦβ１）在细胞生长的负性调节上有重要作用,是肝细胞生长和增殖的强抑制物但肝癌（ＨＣＣ）患者肝组织中ＴＧＦβ１ｍＲＮＡ表达高．现检测ＨＣＣ患者外周血中ＴＧＦβ１ｍＲＮＡ表达,血清ＴＧＦβ１水平,以及肝组织中ＴＧＦβⅡ型受体（ＴＧＦβＲⅡ）的表达．方法ＨＣＣ患者外周血分离单个核细胞（ＰＢＭＣ）,提取总ＲＮＡ,用反转录聚合酶链反应（ＲＴＰＣＲ）技术扩增ＴＧＦβ１ｍＲＮＡ．血清ＴＧＦβ１水平测定用Ｐｒｏｍｅｇａ公司产的试剂盒．肝组织ＴＧＦβＲⅡ表达的分析用原位杂交法．结果ＨＣＣ患者２０例ＰＢＭＣＴＧＦβ１ｍＲＮＡ用ＲＴＰＣＲ检测阳性率达７０％,对照组阴性．ＨＣＣ患者４０例血清ＴＧＦβ１水平（２７５８ｍｇ／Ｌ±８１０ｍｇ／Ｌ）明显高于对照组（８２７ｍｇ／Ｌ±３７２ｍｇ／Ｌ）．原位杂交表明,ＴＧＦβＲⅡ在ＨＣＣ细胞的胞质中有弱表达．结论ＨＣＣ患者ＴＧＦβ１基因在转录水平和翻译水平上均显示表达增强．ＰＢＭＣ有可能代替肝组织用以检测ＴＧＦβ１ｍＲＮＡ的表达应用于基础与临床的研究．     

类风湿关节为中转化生长因子β1和细胞间粘附分子的变化

目的：了解类风湿关节炎（ＲＡ）血清、滑液和滑膜组织中转化生物因β１（ＴＧＦβ１）、细胞间粘附分子－１（ＩＣＡＭ－１）及细胞间粘附分子－３（ＩＣＡＭ－３）的变化。方法：采用ＥＬＩＳＡ及ＡＢＣ免疫组化方法检测ＲＡ患者血清、滑液和滑膜中ＴＧＦβ１、ＩＣＡＭ－１和ＩＣＡＭ－３的浓度和阳性程度。结果：ＲＡ血清中ＴＧＦβ１含量明显升高，ＩＣＡＭ－３含量正常，滑液中ＩＣＡＭ－３含量低于血清含量。ＲＡ血清中ＴＧＦ     

类风湿关节炎滑膜转化生长因子β1表达及病理改变的关系

目的 了解转化生长因子β１（ＴＧＦβ１）在类风湿关节炎（ＲＡ）滑膜组织中的表达及与病理改变的关系，以探讨ＴＧＦβ１在ＲＡ发病中的作用。方法 采用免疫组织化学ＬＳＡＢ法检测了５３例（其中２６例ＲＡ，２０例其他关节炎和７例正常人）滑膜组织中ＴＧＦβ１染色强度及面积的定量分析。结果 ＲＡ滑膜衬里层ＴＧＦβ１阳性细胞数及阳性染色强度、衬里下层染色强度及阳性面积均较正常组和其他关节炎组明显增强（Ｐ〈０．０５）；ＲＡ滑膜衬里层细胞层数、衬里下层炎症细胞的浸润均较对照组明显增多；活动期ＲＡ滑膜衬里层和衬里下层ＴＧＦβ１染色均较静止期明显增强，并且活动期滑膜炎中，慢性活动期、肉芽组织增生期较新近活动期衬里下层染色强度及阳性面积明显升高（Ｐ〈０．０５）。结论 ＲＡ滑膜ＴＧＦβ１蛋白合成、分泌增多，并且增多的ＴＧＦβ１可能与ＲＡ滑     

转化生长因子β1单克隆抗体干预成纤维细胞增殖的体外研究

目的观察抗转化生长因子β１（ＴＧＦβ１）单克隆抗体对成纤维细胞（ＦＢ）增殖及胶原合成的影响。方法将肺泡巨噬细胞（ＡＭ）培养上清液与Ｌ９２９成纤维细胞株共同孵育,并加入不同剂量ＴＧＦβ１单克隆抗体,用３Ｈ胸腺嘧啶掺入法观察ＦＢ增殖及胶原合成的情况。结果抗ＴＧＦβ１单克隆抗体能够抑制ＡＭ条件上清液引起的ＦＢ增殖,在一定剂量范围内（１０μｇ／ｍｌ）呈剂量效应关系。抑制作用在１０μｇ／ｍｌ时达到最大并使ＦＢ培养上清Ⅳ型胶原的合成减少３２％。结论提示抗ＴＧＦβ１单克隆抗体的应用可能为肺纤维化的治疗提供潜在的新途径。     

血吸虫病肝纤维化患者转化生长因子β_1mRNA的水平及其临床意义

目的：研究血吸虫病患者 Ｔ Ｇ Ｆβ１ ｍ Ｒ Ｎ Ａ 水平及其临床意义。方法：用 Ｒ Ｔ Ｐ Ｃ Ｒ 加 ｄｏｔｂｌｏｔ法测定血吸虫病患者 Ｐ Ｂ Ｍ Ｃ中 Ｔ Ｇ Ｆβ１ ｍ Ｒ Ｎ Ａ 水平,与肝硬变和肝癌患者作比较,并研究了部分肝脏组织（肝癌患者１６ 例,肝血管瘤患者正常肝组织 ５ 例）中 Ｔ Ｇ Ｆβ１ ｍ Ｒ Ｎ Ａ 水平与 Ｐ Ｂ Ｍ Ｃ中水平的关系。同时,测定血清中 Ｈ Ａ、 Ｌ Ｎ、 ＣｏｌⅠⅤ和 Ｐ ＣⅢ水平,作为衡量肝纤维化活动与否的指标。结果： Ｐ Ｂ Ｍ Ｃ内 Ｔ Ｇ Ｆβ１ ｍ Ｒ Ｎ Ａ 水平在晚期血吸虫病患者组（ｎ＝ ２１,１２６±０１４）,肝硬变患者组（ｎ＝ １５,２０５±０８１）和肝癌患者组（ｎ＝ ２５,１８３±１２９）均显著高于正常对照组（ｎ＝ １６,０６２±０４０）（ Ｐ＜ ００５）。其中晚期血吸虫病患者组又显著低于肝硬变患者组或肝癌患者组（ Ｐ＜ ００５）,后两组差异无显著性（ Ｐ＞ ００５）。肝组织与 Ｐ Ｂ Ｍ Ｃ内 Ｔ Ｇ Ｆβ１ ｍ Ｒ Ｎ Ａ 水平差别无统计学意义（ Ｐ＞ ００５）。血清 Ｈ Ａ、 ＣｏｌⅣ和 Ｌ Ｎ 异常组的 Ｔ Ｇ Ｆβ１ ｍ Ｒ Ｎ Ａ 水平显著高于正常组（ Ｐ＜ ００５）。结论： Ｐ     

胃粘膜组织幽门螺杆菌感染与TGF-β1表达的关系

目前认为,幽门螺杆菌（Ｈｐ）及其细胞毒素相关蛋白（ＣａｇＡ）可能与胃癌发生相关［１］．转化生长因子β１（ＴＧＦβ１）是重要的细胞生长调控成分,与肿瘤的发生关系密切,本文采用ＰＣＲ技术和Ｈｐ快速尿素酶试验（ＨｐＲＵＴ）同步检测慢性浅表性胃炎（ＣＳＧ）、胃粘膜肠化生（ＩＭ）、不典型增生（Ｄｙｓ）及胃腺癌（ＧＡＣ）组织中Ｈｐ感染情况并检测其ｃａｇＡ基因型,以免疫组化方法分别检测其中ＴＧＦβ１的表达,探讨Ｈｐ、特别是ｃａｇＡ阳性Ｈｐ感染与ＴＧＦβ１表达在胃癌发生过程的作用．１　材料和方法１．１　材…     

TGF-β1及TGF-β1受体在胃癌及癌前病变中的表达

目的探讨ＴＧＦβ１与ＴＧＦβＩ受体（ＲⅠ）在胃癌发生发展中的作用．方法采用免疫组化ＳＰ法检测基本正常胃粘膜（３０例）、肠化生（３０例）、不典型增生（２２例）及胃癌（２５例）中ＴＧＦβ１与ＴＧＦβＲⅠ的表达．结果肠化生、不典型增生及胃癌中ＴＧＦβ１的表达增强而ＴＧＦβＲⅠ表达递减（Ｐ＜００５）;不典型增生组与胃癌组间ＴＧＦβ１表达无显著差异（Ｐ＞００５）;１９例胃癌（７６％）ＴＧＦβＲⅠ表达缺失;ＴＧＦβ１与ＴＧＦβＲⅠ表达与胃癌的浸润深度及淋巴结转移无关,在癌周正常组织与正常粘膜组间亦无显著差异（Ｐ＜００５）．结论ＴＧＦβＲⅠ表达缺失与ＴＧＦβ１过度表达可能参与胃癌的发生     

Octreotide治疗Graves'眼病机理探讨

目的　探讨Ｏｃｔｒｅｏｔｉｄｅ 对细胞因子刺激后的人眼球后成纤维细胞的细胞间粘附分子１（ＩＣＡＭ１） 表达及ＤＮＡ 合成的影响。方法　应用细胞培养技术, 测定加入细胞因子及Ｏｃｔｒｅｏｔｉｄｅ后ＩＣＡＭ１ 表达和ＤＮＡ 合成情况。结果　ＴＮＦα（１０３ Ｕ／ｍｌ） 、γＩＮＦ（１００ Ｕ／ｍｌ） 和ＩＬ１β（１００ Ｕ／ｍｌ） 均能强烈刺激眼球后成纤维细胞表面ＩＣＡＭ１ 表达,而低浓度（ 即药理浓度）Ｏｃｔｒｅｏｔｉｄｅ 能明显地抑制这种刺激作用,但所需浓度及发挥抑制作用的时间各不相同;同样,低浓度Ｏｃｔｒｅｏｔｉｄｅ 也能抑制细胞因子（除ＩＬ１β外） 刺激的ＤＮＡ 合成。结论　Ｏｃｔｒｅｏｔｉｄｅ 治疗Ｇｒａｖｅｓ’眼病的作用途径部分是通过抑制细胞因子的作用,使ＩＣＡＭ１ 表达及ＤＮＡ 合成减少,它可能具有免疫调节的特性。     

40-氧-(2-羟乙基)-雷帕霉素预防大鼠移植肾慢性排斥反应实验研究

目的　研究４０氧（２羟乙基）雷帕霉素（ＳＤＺＲＡＤ） 对大鼠移植肾慢性排斥反应的预防作用。方法　应用显微外科技术制作移植肾慢性排斥反应大鼠模型,将受体大鼠随机分为两组。治疗组大鼠接受ＳＤＺＲＡＤ０．５ ｍｇ·ｋｇ－１·ｄ－ １ 灌胃,对照组接受安慰剂。移植后每４ 周行２４ 小时尿蛋白定量测定,第２４ 周处死大鼠,对移植肾组织行组织学、免疫组化及反转录聚合酶链反应（ＲＴＰＣＲ） 检测。结果　治疗组大鼠的蛋白尿、肾小球硬化、血管内膜增厚、单核巨噬细胞和淋巴细胞浸润等病变程度均较对照组为轻,黏附分子ＩＣＡＭ１、ＶＣＡＭ１ 和生长因子ＴＧＦβ１、ＰＤＧＦＡＡ 基因的表达减少。结论　ＳＤＺＲＡＤ可预防同种移植肾慢性排斥反应。黏附分子和生长因子基因表达的减少,可能是ＳＤＺＲＡＤ预防同种移植肾慢性排斥反应机制中的重要环节。     

Expression of basic fibroblast growth factor in synovial tissues from patients with rheumatoid arthritis: detection by immunohistological staining and in situ hybridisation.

OBJECTIVE--The distribution and production of basic fibroblast growth factor (bFGF) was examined on the synovium from patients with rheumatoid arthritis (RA) and osteoarthritis (OA). METHODS--The localisation of bFGF was determined by an immunohistochemical staining procedure using anti-bFGF monoclonal antibody. The expression of bFGF mRNA was detected by nonradioactive in situ hybridisation using bFGF antisense oligo DNA. RESULTS--The bFGF was found in the synovial lining cell, sublining stromal fibroblast-like cells, and vascular endothelial cells from patients with RA and OA. Little or no bFGF was found in non-inflamed synovium. Immunostaining of bFGF in the synovial cells was more extensive and intense in synovium of patients with RA than that of patients with OA. The nuclei of the synovial lining cell layer were also immunostained. These nuclear staining were more intense in the lining cell layer from RA patients with moderate or severe proliferation of synovial cells than in RA patients with mild proliferation. The bFGF mRNA was also detected in the synovial lining cell layer of the inflamed synovium. CONCLUSION--The synovial lining cells produced bFGF. The proliferation of synovial cells in the inflamed joints may be the results of stimulation by the bFGF in autocrine manner.     

Differential Distribution of Intercellular Adhesion Molecules (ICAM–1, ICAM–2, and ICAM–3) and THE MS–1 Antigen in Normal and Diseased Human Synovia

Objective. Cellular adhesion and differentiation molecules (CAMs) may play a role in the recruitment and retention of inflammatory cells into rheumatoid arthritis synovial tissue (RA ST). In order to determine if certain CAMs are up-regulated in RA ST compared with normal ST, we studied the distribution of intercellular adhesion molecules (ICAMs) 1, 2, and 3 in ST. We also studied the MS-1 antigen since it is preferentially expressed on discontinuous endothelia, such as those found in RA ST; MS-1 is also expressed differentially upon cytokine activation of cells in vitro or in pathologic conditions in situ. Thus, we postulated a possible similarity between MS-1 and ICAM-1 expression in inflamed ST. Methods. Immunohistochemical analysis was used to determine the distribution of ICAMs and MS-1 in ST from 10 patients with RA, 10 with osteoarthritis (OA), and 4 normal individuals. Results. ICAM-1 expression was found on significantly more RA ST endothelial cells compared with normal cells, as well as on RA ST macrophages and lining cells. ICAM-2, also found on endothelial cells, showed no differential staining pattern. ICAM-3 was present on RA ST macrophages and lining cells as well as on some RA and OA endothelial cells. The MS-1 antigen was present on most RA and OA ST endothelia, lining cells, and macrophages. ICAM-1 expression and MS-1 expression in the lining layer were positively correlated in both RA and OA. Conclusion. ICAM-1, while found mainly on endothelial cells, is up-regulated on RA ST macrophages and lining cells, suggesting a role for these cells in the infiltration and tissue damage seen in the RA ST. ICAM-3, which is present mainly on normal resting leukocytes but not on normal endothelium, is expressed by some diseased ST leukocytes and endothelial cells. MS-1 is also found on the RA ST specialized, fenestrated endothelium, on macrophages, and in the lining layer. These results suggest that the differential expression of ICAMs and MS-1 in RA ST compared with normal ST might play a special role in the pathogenesis of RA.     

Dynamics of interleukin (IL)-18 in serum, synovial fluid and synovial membrane in the patients with rheumatoid arthritis]

OBJECTIVES: IL-18 is a novel cytokine that plays an important role in the Th1 response. The aim of this study is to investigate the dynamics of IL-18 in serum, synovial fluid and synovial membrane in the patients with rheumatoid arthritis. MATERIALS AND METHODS: The serum, synovial fluid and synovial membrane were obtained from RA patients at operation. The levels of IL-18 in the serum and synovial fluid were measured by ELISA. We then examined the expression of IL-18 in synovial tissues using anti-human IL-18 monoclonal antibody in immunohistochemical study. RESULTS: The levels of IL-18 in serum and synovial fluid in RA patients were 193.7 +/- 109.7 pg/ml and 258.8 +/- 238.0 pg/ml, respectively. Compared with OA patients and normal volunteers, the level of IL-18 in RA patients was higher in both serum and synovial fluid. (P < 0.05) In synovial membrane, the cells positive for anti IL-18 antibody were confirmed not only in RA (n = 26) but also in OA (n = 7) patients. The positive cells were the synovial lining cells, macrophages, fibroblasts and endothelial cells. However, a large number of positive cells were demonstrated in synovial tissues in RA compared with OA patients.     

Comparative analysis of cathepsin l,cathepsin d,and collagenase messenger rna expression in synovial tissues of patients with rheumatoid arthritis and osteoarthritis,by in situ hybridization

Objective. To compare the expression of cathepsin L, cathepsin D, and collagenase messenger RNA (mRNA) in synovial specimens from patients with rheumatoid arthritis (RA) and osteoarthritis (OA). Methods. The expression of cathepsins L and D as well as collagenase mRNA in synovial tissues from 8 patients with RA, 6 patients with OA, and 2 patients with noninflamed joints was evaluated using in situ hybridization with digoxigenin-labeled RNA probes. Results. Both RA and OA synovial tissue expressed cathepsins L and D as well as collagenase mRNA. The expression of the cathepsins was markedly higher in interstitial regions and, to some extent, in perivascular infiltrates of RA synovial tissue compared with OA specimens. Conclusion. Cathepsins L and D mRNA are expressed differently in RA and OA synovial tissues, supporting the concept that these enzymes may contribute to the influx of mononuclear cells into RA synovium. Moreover, the data reveal that the expression of collagenase and cathepsins in RA and OA synovial lining is otherwise largely similar, and suggest that the adhesion of synovial cells to cartilage mediates the invasive destructive process in RA.     

RNA released from necrotic synovial fluid cells activates rheumatoid arthritis synovial fibroblasts via Toll-like receptor 3

OBJECTIVE: To assess the expression of Toll-like receptor 3 (TLR-3) protein in synovial tissues and cultured synovial fibroblasts obtained from patients with rheumatoid arthritis (RA) and osteoarthritis (OA) and to investigate the consequences of stimulation of cultured synovial fibroblasts with TLR-3 ligands. METHODS: TLR-3 expression in synovial tissues was determined by immunohistochemistry and immunofluorescence, and expression in cultured RA synovial fibroblasts (RASFs) was determined by fluorescence-activated cell sorting and real-time polymerase chain reaction techniques. TLR-3 signaling was assessed by incubating RASFs with poly(I-C), lipopolysaccharide, palmitoyl-3-cysteine-serine-lysine-4, or necrotic synovial fluid cells from RA patients in the presence or absence of hydroxychloroquine or Benzonase. Subsequent determination of interferon-beta (IFNbeta), CXCL10, CCL5, and interleukin-6 (IL-6) protein production in the culture supernatants was performed by enzyme-linked immunosorbent assays. RESULTS: TLR-3 protein expression was found to be higher in RA synovial tissues than in OA synovial tissues. TLR-3 expression was localized predominantly in the synovial lining, with a majority of the TLR-3-expressing cells coexpressing fibroblast markers. Stimulation of cultured RASFs with the TLR-3 ligand poly(I-C) resulted in the production of high levels of IFNbeta, CXCL10, CCL5, and IL-6 protein. Similarly, coincubation of RASFs with necrotic synovial fluid cells from patients with RA resulted in up-regulation of these cytokines and chemokines in a TLR-3-dependent manner. CONCLUSION: Our findings demonstrate the expression of TLR-3 in RA synovial tissue and the activation of RASFs in vitro by the TLR-3 ligand poly(I-C) as well as by necrotic RA synovial fluid cells, and indicate that RNA released from necrotic cells might act as an endogenous TLR-3 ligand for the stimulation of proinflammatory gene expression in RASFs.