In vivo and in vitro expression of adhesion molecules by peripheral blood lymphocytes from patients with primary Sjogren's syndrome: culture-associated enhancement of LECAM-1 and CD44

The interaction of adhesion receptors on lymphocytes with their ligands over endothelial cells provides the mechanism by which lymphocytes infiltrate target tissues in autoimmune diseases. Primary Sjogren's syndrome (SS) is associated with lymphocytic infiltration in exocrine glands. The aim of this study was to examine levels of expression of adhesion molecules by peripheral blood lymphocytes from patients with SS (before and after stimulation). Peripheral blood lymphocytes from 16 patients with primary SS and from 15 controls were stained directly or cultured for 72 h with and without phytohaemagglutinin (PHA). Indirect immunofluorescence with monoclonal antibodies and flow cytometry were used. The following molecules were detected in patients before culture: CD18 (mean percentage 94%), CD11a (94%), CD11b (39%), CD54 (23%), CD58 (62%), CD44 (Hermes-1; 82%), CD49-d (VLA-4; 80%), CD25 (11%) and LECAM-1 (62%). After stimulation with PHA, there was an increase in the levels of CD18 (2.5-fold), CD11a (2.3-fold), CD54 (10.2-fold), CD58 (2.5-fold), CD44 (2.4-fold), CD49d (3.4-fold) and CD25 (62-fold) on lymphocytes from both patients and controls. The number of positive cells and level of expression did not differ from the controls, except in the case of unstimulated, cultured lymphocytes in which the levels of CD44 and LECAM-1 were increased more in patients than in normal controls. The increase in the level of in vitro expression of CD44 (P< 0.05) and LECAM-1 (P< 0.002) on lymphocytes from patients with primary SS reached statistical significance when compared to similarly cultured lymphocytes from controls. The regulation of LECAM-1 and CD44 expression in patients with SS may contribute to the immunopathogenesis of this desease by increased tethering or rolling (early adhesive events) of lymphocytes over endothelial cells. Such molecules may be involved in lymphocytic homing to tissues in SS.     

TLR4激动上调内皮细胞氧化低密度脂蛋白受体LOX-1表达

目的近年研究表明介导先天免疫反应的受体Toll样受体4(TLR4)参与了动脉硬化的发生发展。业已证明氧化低密度脂蛋白受体LOX-1介导内皮细胞活化和功能失调，激发炎症过程，在动脉粥样硬化的发生和发展中起着极为重要作用。本研究观察TLR4激动是否调节内皮细胞LOX-1表达。方法应用脂多糖(LPS)刺激体外培养的人脐静脉内皮细胞(HUVECs)24h。采用RT—PCR和流式细胞术分别检测TLR4、LOX-1 mRNA和蛋白表达水平。为了观察转录因子NF—kB在调节LOX-1表达中的作用，应用NF-kB特异性抑制剂咖啡酸苯乙酯(CAPE)预处理细胞，然后以LPS刺激，检测LOX-1 mRNA和蛋白变化。结果LPS(10～1000ng／mL)上调HUVECs TLR4和LOX-1 mRNA表达，LPS(1000ng／mL)上调TLR4和LOX-1蛋白表达，CAPE(20μg／mL)可抑制LPS介导的LOX-1表达上调。结论TLR4／NF-kB信号途径可能通过上调内皮细胞LOX-1表达参与动脉粥样硬化的发生及发展。     

The expression of chemokine receptor CXCR3: relevance to disease activity of rheumatoid arthritis

 CXC chemokine receptor 3 (CXCR3) is selectively expressed on T helper 1 (Th1) type T cells and has been shown to be responsible for Th1-dominant immune responses. In this study, we analyzed the expression of CXCR3 on peripheral blood T lymphocytes of patients with rheumatoid arthritis (RA) by FACS analysis using antihuman CXCR3 monoclonal antibody and determined the clinical relevance in this disease. Significantly higher expression of CXCR3 was found on peripheral blood CD4+ T lymphocytes of RA patients than healthy controls. The CXCR3 expression in RA patients with a high erythrocyte sedimentation rate was significantly higher than in those with a low erythrocyte sedimentation rate. Moreover, we found that the CXCR3 expression in RA patients with long-term disease duration was significantly higher than in those with short-term disease. On the other hand, CC chemokine receptor 4 (CCR4), which was shown to be selectively expressed on Th2-type T cells, was expressed at low levels in RA patients as well as in healthy controls. The serum level of interleukin (IL)-18 in RA patients was higher than that in healthy controls, although there was no statistically significant difference. This study suggests that the Th1 immune response is predominant in RA and that CXCR3 may have relevance in regard to the disease course in RA patients. Received: January 28, 2002 / Accepted: August 14, 2002     

反式载脂蛋白A1模拟肽R-D4F对内皮祖细胞黏附及迁移功能的影响

目的探讨反式载脂蛋白A1模拟肽R-D4F对人内皮祖细胞黏附及迁移功能的影响。方法取新鲜人脐静脉血,采用密度梯法从中分离、诱导培养人内皮祖细胞,并使用Dil-ac-LDL和FITC-UEA-1染料进行双荧光染色以及流式细胞术进行鉴定。分别给于牛血清白蛋白、顺式D-4F(50μg/ml)及R-D4F(5~50μg/ml)处理后,采用黏附能力测定实验及Transwell小室观察内皮祖细胞黏附及迁移能力的变化。结果与牛血清白蛋白对照组相比,D-4F显著增强内皮祖细胞的黏附及迁移功能。相似的是,R-D4F呈剂量依赖性地增强内皮祖细胞的黏附与迁移。结论 R-D4F具有与D-4F相似的生物学作用,可促进人内皮祖细胞的黏附及迁移。     

外周血中TLR4、TIRAP表达与脓毒症相关性研究

目的探讨外周血中TOLL样受体4(TLR4)及TOLL/白介素-1受体相关蛋白(TIRAP)水平与脓毒症严重程度的相关性。方法 30例正常健康体检者为正常组,53例脓毒症患者,进行APACHEⅡ评分,APACHEⅡ≤20患者40例为脓毒症低评分组,APACHEⅡ20患者13例为脓毒症高评分组。所有研究对象应用酶联免疫吸附试验(ELISA)检测静脉血清总TLR4、TIRAP浓度。结果血清中TLR4及TIRAP浓度:正常组为0.886±0.058 ng/ml及5.216±0.410 ng/ml、脓毒症低评分组为2.253±0.379 ng/ml及9.540±2.294ng/ml、脓毒症高评分组为4.494±0.709 ng/ml及19.206±1.755 ng/ml,脓毒症低评分组及脓毒症高评分组与正常组相比均有显著性差异(P均=0.000),脓毒症低评分组与脓毒症高评分组相比存在显著性差异(P均=0.000)。以APACHEⅡ评分等于20分为界,即脓毒症低评分组为低危患者,脓毒症高评分组为高危患者,来区分脓毒症严重程度。经Pearson相关分析,TLR4浓度与脓毒症严重程度呈正相关(r=0.931,P0.05),TIRAP浓度与脓毒症严重程度也呈正相关(r=0.972,P0.05),TLR4浓度与TIRAP浓度呈正相关(r=0.936,P0.05)。结论脓毒症患者血清总TLR4、TIRAP水平可以预测及判断脓毒症严重程度。     

促血管生成素-1对糖尿病大鼠肾脏色素上皮衍生因子和血小板反应蛋白-1基因表达的影响

目的 观察色素上皮衍生因子（PEDF）、血小板反应蛋白-1（TSP-1）在糖尿病大鼠肾脏的表达变化,探讨促血管生成素-1（Ang-1）对上述因子的影响. 方法 将雄性SD大鼠分正常对照（NC）组、DN组、空载处理（BV）组、Ang-1处理（AV）组.采用STZ腹腔注射诱导大鼠DN模型.成模8周后尾静脉注射Ang-1腺病毒载体.多时点检测24 hUAlb和肾组织PEDF、TSP-1蛋白及mRNA表达水平.结果 DN、BV、AV组24 hUAlb均升高,其中AV组于20周后降低（P＜0.05）.DN、BV、AV组肾组织PEDF蛋白及mRNA表达下调,TSP-1表达上调（P＜0.05）,其中AV组12周后肾组织PEDF和TSP-1mRNA及蛋白表达变化较DN、BV组改善明显（P＜0.05）. 结论 糖尿病大鼠肾脏PEDF、TSP-1异常表达参与DN发生发展,给予Ang-1可改善糖尿病大鼠肾脏PEDF、TSP-1异常表达.     

HIV/AIDS患者血清IL-18浓度变化与辅助受体CCR5/CXCR4表达的关系研究

目的 研究人类免疫缺陷病毒／获得性免疫缺陷综合征（HIV／AIDS）患者血清白细胞介质（IL）-18浓度的变化，及其与病毒载量、CD4^＋T细胞表面辅助受体CCR5／CXCR4表达的相关性。方法 应用酶联免疫吸附试验（ELISA）对42例HIV／AIDS患者及12例正常对照者的血清IL-18浓度进行定量检测，分析其变化，并对其变化与病毒载量的相关性进行分析。应用流式细胞仪检测全部HIV／AIDS患者CD4^＋ T细胞表面辅助受体CCR5／CXCR4的表达情况，并分析其与病毒载量及血清IL-18浓度变化之间的相关性。结果 42例HIV／AIDS患者血清平均IL-18浓度显著高于正常对照组（P〈0．01），且按美国疾病控制与预防中心（CDC）1993年标准，CDCC期显著高于A期（P〈0．01）。所有患者血清IL-18浓度变化与HIV病毒载量之间呈正相关（P〈0．05），与辅助受体CX-CR4的表达也呈正相关（P〈0．01）。HIV病毒载量与辅助受体CXCR4的表达具有正相关性（P〈0．05）。结论 HIV／AIDS患者体内持续增长的IL-18加剧了其免疫功能的紊乱和低下，IL-18可能与疾病本身的致病机制有关。     

脑胶质瘤中MAGE-D4和CORO1C mRNA表达水平及其临床意义

目的 分析脑胶质瘤中黑色素瘤相关抗原(MAGE)-D4和肌动蛋白结合蛋白CORO1C mRNA的表达水平,探讨二者的相关性及临床意义.方法 通过基因表达谱交互式分析(GEPIA)数据库收集脑胶质瘤样本数据681例[低级别脑胶质瘤(LGG)518例,胶质母细胞瘤(GBM)163例],正常脑组织样本数据207例,比较其MA...     

AQP4与PKC在大鼠CIR后脑水肿模型中的变化及作用

目的通过研究大鼠局灶性脑缺血再灌注后水通道蛋白4（AQP4）、蛋白激酶C（PKC）表达水平的变化,来探讨两者与脑水肿之间的关系。方法通过线栓法复制大鼠大脑中动脉缺血（MCAO）再灌注模型,动物随机分为正常对照组（Norm）、假手术组（Sham）、缺血再灌注组（CIR）。参照Garcia JH法进行神经功能缺损评分、用Elliot法检测脑组织含水量、免疫组化法检测AQP4及PKC的表达。结果 CIR后6h即有神经功能缺损,脑含水量在CIR12h明显升高,1~3d时达到高峰;AQP4表达在早期水平降低,从12h逐渐升高,至CIR 24h明显升高,3d达高峰;PKC在CIR后6h开始持续性升高,至第5d仍保持较高水平。结论早期的细胞源性脑水肿可能和PKC的升高有关,后期的血管源性脑水肿可能和AQP4的升高相关。     

Glutathione S-Transferase M1 Gene Polymorphisms are Associated with Cardiac Iron Deposition in Patients with β-Thalassemia Major

Patients with β-thalassemia (thal) major are subject to peroxidative tissue injury by iron overload. Glutathione S-transferases work as antioxidants, and their activity is determined genetically. In this study, we used multiplex polymerase chain reaction (m-PCR) to analyze polymorphisms of two endogenous antioxidant agents, glutathione S-transferase M1 (GSTM1) and glutathione S-transferase T1 (GSTT1), and to determine their roles in 41 patients with β-thal major. Our results showed that the GSTM1 and GSTT1 null genotypes were not associated with any incidence of endocrine dysfunction (including diabetes mellitus, hypogonadism, hypothyroidism, and growth hormone deficiency), liver function, or impaired left ventricular ejection fraction (LVEF). The GSTM1 null genotype, but not the GSTT1 null genotype, was associated with a decreased signal intensity ratio on cardiac magnetic resonance imaging (MRI). Our results suggest that genetic variations of the GSTM1 enzyme are associated with cardiac iron deposition in patients with β-thal major.     

钙调素依赖蛋白激酶Ⅱ调控SSH-1L活性的作用

目的 探讨钙(Ca~(2+))/钙调素依赖蛋白激酶Ⅱ(CaMKⅡ)对Slingshot-1L(SSH-1L)活性的调控作用.方法 用脂质体法将含Myc-SSH-1L的重组质粒转染至MG63细胞,经不同浓度的钙离子载体A23187诱导10 min,蛋白印迹实验,观察P-cofilin、P-SSH1L、P-CaMK Ⅱ水平的变化;体内及体外实验检测CaMKⅡ对SSH-1L的磷酸化及活性调控作用.结果 当 A23187浓度为1 μmol/L时CaMK Ⅱ活性达到了最大,同时此浓度下P-cofilin水平以及SSH-1L的978位点丝氨酸的磷酸化水平升高.体外实验证实CaMK Ⅱ可使SSH-1L(WT)磷酸化,但是对其突变体SSH-1L(2 SA)无作用;同时CaMK Ⅱ明显抑制了SSH1L(WT)的活性,但对SSH-1L(2 SA)的活性无作用.结论 CaMKⅡ对SSH-1L的活性具有明显调控作用,且与SSH-1L的丝氨酸位点密切相关.     

In Vitro and In Vivo Characterization of a Pigeon Paramyxovirus Type 1 Isolated from Domestic Pigeons in Victoria,Australia 2011

Significant mortalities of racing pigeons occurred in Australia in late 2011 associated with a pigeon paramyxovirus serotype 1 (PPMV-1) infection. The causative agent, designated APMV-1/pigeon/Australia/3/2011 (P/Aus/3/11), was isolated from diagnostic specimens in specific pathogen free (SPF) embryonated eggs and was identified by a Newcastle Disease virus (NDV)-specific RT-PCR and haemagglutination inhibition (HI) test using reference polyclonal antiserum specific for NDV. The P/Aus/3/11 strain was further classified as PPMV-1 using the HI test and monoclonal antibody 617/161 by HI and phylogenetic analysis of the fusion gene sequence. The isolate P/Aus/3/11 had a slow haemagglutin-elution rate and was inactivated within 45 min at 56 °C. Cross HI tests generated an R value of 0.25, indicating a significant antigenic difference between P/Aus/3/11 and NDV V4 isolates. The mean death time (MDT) of SPF eggs infected with the P/Aus/3/11 isolate was 89.2 hr, characteristic of a mesogenic pathotype, consistent with other PPMV-1 strains. The plaque size of the P/Aus/3/11 isolate on chicken embryo fibroblast (CEF) cells was smaller than those of mesogenic and velogenic NDV reference strains, indicating a lower virulence phenotype in vitro and challenge of six-week-old SPF chickens did not induce clinical signs. However, sequence analysis of the fusion protein cleavage site demonstrated an ¹¹²RRQKRF¹¹⁷ motif, which is typical of a velogenic NDV pathotype. Phylogenetic analysis indicated that the P/Aus/3/11 isolate belongs to a distinct subgenotype within class II genotype VI of avian paramyxovirus type 1. This is the first time this genotype has been detected in Australia causing disease in domestic pigeons and is the first time since 2002 that an NDV with potential for virulence has been detected in Australia.