SLE患者外周血内皮祖细胞数量及生物学功能的研究

目的 探讨SLE患者外周血内皮祖细胞数量和功能的变化。方法 女性SLE患者及正常人对照各20例,取外周血20ml,密度梯度离心法分离单一核细胞,采用CD133/CD34、CD133/VEGFR2双荧光标记鉴定细胞,CD34/CD133/VEGFR2三荧光标记流式检测EPC细胞数量,采用MTT比色法、改良的millicell小室和黏附能力测定实验观察EPC的增殖、迁移和黏附能力。结果 SLE患者外周血内皮祖细胞数量较正常人对照明显下降（4.49% ± 1.66%比20.81% ± 4.14%,P < 0.01）,其增殖（23.11 ± 3.16比35.65 ± 1.74）、迁移（12.00 ± 2.12个细胞/视野比23.60 ± 3.0个细胞/视野）、黏附能力（22.43 ± 4.43个细胞/视野36.43 ± 3.69个细胞/视野）均有所下降（P < 0.01）。结论 SLE患者外周血内皮祖细胞数量减少、功能受损。     

结缔组织疾病

Toll样受体7及I型干扰素通路在系统性红斑狼疮中的作用研究;自体外周血纯化CD34^＋细胞移植对系统性红斑狼疮患者外周血CD4^＋CD25^＋T细胞表达的影响;IL-18及IL-13在系统性红斑狼疮患者肾组织中的表达及意义;首诊系统性红斑狼疮住院患者ANA、抗ds-DNA与病情活动的研究;中西药合用治疗系统性红斑狼疮34例观察     

SLE患者外周血中CD4+CD25+CD127low/-标记的调节性T细胞与临床表现和实验室指标相关性研究

目的:探讨CD4+CD25+CD127low/-标记的调节性T细胞(Treg)在系统性红斑狼疮发病机制中的作用.方法:用流式细胞仪检测45例系统性红斑狼疮(SLE)患者和45例年龄、性别相匹配的健康志愿者外周血CD4+T细胞中Treg的百分比,同时分析SLE患者外周血中的CD4+CD25+CD127low/-标记的Tr...     

SLE患者外周血造血干/祖细胞的检测及临床意义

目的 探讨SLE患者外周血CD34+造血干/祖细胞（HSC/HPC）数目与CD34膜表达的变化。方法 采用异硫氰酸荧光素标记抗体,以流式细胞仪检测30例SLE患者和14例正常人外周血CD34+ HSC/HPC,分析CD34+ HSC/HPC细胞占全部淋巴细胞的百分比和CD34平均荧光强度,并结合临床资料进行相关性分析。结果 活动期和稳定期SLE患者外周血CD34+ HSC/HPC细胞占淋巴细胞百分比分别为（0.15 ± 0.10）%和（0.09 ± 0.07）%,低于正常人对照组［（0.37 ± 0.17）%,F = 17.18,P < 0.01］,而活动期和稳定期SLE患者差异无统计学意义（t = 1.51,P > 0.05）;活动期SLE患者外周血CD34抗原的平均荧光强度为41.35 ± 19.24,高于正常人对照组（27.43 ± 7.57,F = 3.13,P < 0.05）,而稳定期SLE患者与正常人对照组差异无统计学意义（F = 3.13,P > 0.05）。外周血CD34+ HSC/HPC细胞百分比与血清IgG水平呈负相关（r = -0.588,P < 0.01）,与SLE疾病活动指数（SLEDAI）、补体、抗dsDNA抗体、抗C1q抗体、抗核小体抗体等无统计学相关性。结论 SLE患者外周血CD34+ HSC/HPC细胞数减少,并且CD34抗原表达增加,提示SLE患者HSC/HPC功能存在异常,可能参与SLE的发病。     

系统性红斑狼疮患者外周血中T细胞CD4CD25的表达

目的了解CD4,CD25在系统性红斑狼疮患者外周血T细胞中的表达水平及其临床意义。方法用流式细胞术检测30例正常人和30例活动期系统性红斑狼疮(SLE)患者外周血中T细胞CD4,CD25的表达水平,根据CD25表达荧光强度>100者为T细胞CD4+CD25+bright,并与SLE活动指数评分(SLEDAI)进行相关分析。结果SLE患者外周血T细胞CD4+CD25+表达率为(8.82±2.98)%,显著低于对照组(12.24±5.71)%(P<0.05);患者组T细胞CD4+CD25bright表达率为(1.56±0.76)%,低于对照组(2.23±1.22)%(P<0.05),差异有统计学意义;SLE患者外周血T细胞CD4+CD25+、T细胞CD4+CD25bright表达率与SLEDAI评分两者之间无相关性;r分别为-0.156,-0.153,P均>0.05)。结论SLE患者外周血中T细胞CD4+CD25+减少,其可能在发病机制中起一定的作用,低水平T细胞CD4+CD25bright在SLE发病中的确切作用机制有待进一步阐明。     

系统性红斑狼疮患者血管内皮祖细胞测定

目的 探讨系统性红斑狼疮（SLE）患者外周血中血管内皮祖细胞的变化。 方法 IL ACL-9000型血液凝固仪测定82例SLE患者和50例健康对照vW因子抗原含量（vWF:Ag）。流式细胞仪分析外周血中内皮祖细胞（endothelial progenitor cells,EPC）和循环内皮细胞（circulating endothelial cell,CEC）的水平。用线性相关分析检测vWF:Ag、CEC、EPC等指标之间的相关性。 结果 SLE活动期患者外周血每20万个单一核细胞中CD34+细胞、CD133+细胞、CD34+CD133+双阳性细胞数（35.4 ± 16.7、86.5 ± 32.1、361.3 ± 176.4）分别高于稳定期患者（17.1 ± 10.9、28.7 ± 21.5、107.2 ± 44.3）和健康人（13.8 ± 9.6、11.2 ± 5.5、92.3 ± 50.5）,差异均有统计学意义（P值均 < 0.01）。与稳定期患者比较,SLE活动期患者外周血每20万个单一核细胞中EPC数（361.3 ± 176.4）和vWF:Ag水平（438.9 ± 205.3）%增高,差异有统计学意义（P值均 < 0.01）;CEC差异无统计学意义（P > 0.05）。稳定期患者各项指标与对照组间无统计学意义（P值均 > 0.05）。SLE患者活动期EPC与vWF:Ag呈正相关（r = 0.67,P < 0.01）,与CEC无统计学相关性（P > 0.05）。 结论 SLE患者外周血中EPC数量与血管损伤标志物（vWF:Ag）水平密切相关,表明EPC数量改变可以作为评价病情活动的标志物之一。     

磷脂酰肌醇3-激酶γ抑制剂AS605240对系统性红斑狼疮患者外周血T细胞增殖、黏附和趋化的调控研究

目的:研究磷脂酰肌醇3-激酶γ(PI3Kγ)抑制剂AS605240对系统性红斑狼疮(SLE)患者外周血T淋巴细胞增殖、黏附和趋化的影响,并初步探讨其机制。方法:搜集SLE患者35例,健康人17例为正常组,外周血T细胞分离采用Rosettsep T细胞提取试剂盒;四甲基偶氮唑蓝(MTT)比色法检测细胞增殖,Transwell小室检测T细胞趋化性,丝氨酸/苏氨酸蛋白激酶(AKT)活性检测采用免疫印迹法。结果:AS605240显著抑制SLE患者外周血T细胞体外增殖、黏附活性,并呈浓度依赖关系,当AS605240浓度达到10~(-5)mol/L时差异有统计学意义(P0.01),AS605240显著抑制SLE患者外周血T细胞体外趋化活性(P0.01);SLE患者外周血T细胞AKT磷酸化水平显著高于正常组(P0.05),AS605240处理组T细胞的AKT活性显著降低(P0.05)。结论:P13Kγ抑制剂AS605240显著降低SLE患者外周血T细胞增殖、趋化和黏附活性,与抑制AKT异常活化相关。     

结缔组织疾病

SLE患者血清IL-15水平检测及其意义;白介素-10启动子基因多态性与系统性红斑狼疮相关性研究;血清基质金属蛋白酶-3、-9的检测及其与系统性红斑狼疮活动性的关系;阴虚体质系统性红斑狼疮患者HSP70基因多态性的研究;泌乳素对系统性红斑狼疮患者外周血单个核细胞表面CD40／CD154表达的影响.     

SLE患者外周血中CD4＋CD25＋CD127^low/-标记的调节性T细胞与临床表现和实验室指标相关性研究

目的：探讨CD4＋CD25＋CD127low/-标记的调节性T细胞（Treg）在系统性红斑狼疮发病机制电的作用。方法：用流式细胞仪检测45例系统性红斑狼疮（SLE）患者和45例年龄、性别相匹配的健康志愿者外周血CD4＋T细胞中Treg的百分比,同时分析SLE患者外周血中的CD4＋CD25＋CD127low/-标记的Treg与其临床表现、实验室指标的相关性。结果：SLE患者外周血CD4＋T细胞中Treg百分比与健康对照组比较差异无统计学意义。在SLE患者中,Treg的百分比与抗核小体抗体呈正相关（r=0．299,P=0．046）,有光敏感的SLE患者中Treg较无光敏感组增高（P=0．017）,余均无统计学差异。结论：SLE患者外周血中以CD4＋CD25＋CD127low/-标记的Treg中可能因含有部分无免疫抑制活性的效应性T细胞而特异性差,因此可能不适合用于临床免疫抑制治疗。     

系统性红斑狼疮治疗前后外周血共刺激分子表达的变化和意义

目的探讨共刺激分子表达水平在系统性红斑狼疮(SLE)患者治疗前后的变化及其意义。方法采用免疫荧光标记流式细胞仪检测技术,对SLE患者和正常人外周血单个核细胞膜表面的共刺激分子表达水平进行测定分析。结果①与正常人相比,SLE患者CD28分子表达显著降低(P<0.01),其中双标记CD4+CD28+、CD8+CD28+T细胞均显著减少(P<0.01),4-1BB分子表达显著增高(P<0.01);②患者CD28分子、CD4+CD28+及CD8+CD28+T细胞与SLE病情活动指数(SLEDAI)呈负相关(分别为P<0.01、P<0.01、P<0.05),4-1BB分子与SLEDAI呈正相关(P<0.01);③患者治疗后CD28分子、CD4+CD28+及CD8+CD28+T细胞随病情缓解而上升(P<0.01),4-1BB分子随病情缓解而下降(P<0.01)。结论共刺激分子表达异常在SLE发病机制中可能具有重要作用,其中CD28分子、4-1BB分子、CD4+CD28+及CD8+CD28+T细胞对监测病情活动和判断疗效可能具有重要意义。     

Altered circulating endothelial progenitor cells in patients with keloid

Evidence has suggested that vascular endothelial growth factor (VEGF), a crucial growth factor in regulating endothelial progenitor cells (EPCs), plays a central role in keloid formation. However, the levels of circulating EPCs in patients with keloid have not yet been explored. The aim of this study was to determine the number of circulating EPCs in patients with keloid. Circulating EPCs (defined as CD45? CD34+CD133+VEGFR2+cells) and VEGF levels from 39 patients with keloid and 22 healthy controls (HCs) were assessed by flow cytometry and ELISA, respectively. EPCs were detectable in the peripheral blood of patients with keloid. The number of circulating EPCs and the levels of plasma VEGF were significantly higher in patients with keloid than in HCs. However, no correlation was found between the number of circulating EPCs and the serum VEGF levels. This study provides the first evidence that EPCs are increased in the peripheral blood of patients with keloid. Understanding the roles of EPCs in keloid fromation may lead to the development of novel therapeutic strategies for keloid.     

系统性红斑狼疮患者CD62P、CD63表达与SLEDAI相关性分析

目的:观察系统性红斑狼疮(SLE)患者外周血血小板膜蛋白CD62P、CD63的表达,并探讨CD62P、CD63与SLE疾病活动指数(SLEDAI)之间的相关性。方法:采用流式细胞术对10例SLE患者外周血CD62P、CD63进行检测分析,以20例正常人作为对照。同时对该10例SLE患者进行SLEDAI评分。结果:SLE患者CD62P、CD63表达较对照组显著增高(P<0.01),CD62P表达与SLEDAI之间具有显著相关性(r=0.840,P=0.002)。结论:CD62P、CD63在SLE患者外周血中表达显著增高,CD62P可作为评估SLE疾病活动程度指标。     

High levels of endothelial progenitor cells and circulating endothelial cells in patients with Behcet’s disease and their relationship to disease activity

Background: Behcet’s disease is a multisystemic vasculitis, associated with vascular endothelial dysfunction. Currently, the prognosis is unpredictable, because there is still no valid laboratory marker indicating the disease activity in Behcet’s disease. Endothelial progenitor cells and circulating endothelial cells are newly introduced hematological markers which are presumed to take part in the pathogenesis of vasculitis.Objectives: To evaluate the levels of endothelial progenitor cells and subtypes and circulating endothelial cells in patients with Behcet’s disease and to describe their relationship with the disease activity.Methods: A total of 45 patients with Behcet’s disease and 28 healthy controls were included in the study. Endothelial progenitor cells (CD34+CD133+KDR+ as early endothelial progenitor cells and CD34+KDR+ as late endothelial progenitor cells), and circulating endothelial cells (CD34+CD133+) were measured by flow cytometry.Results: The mean plasma level of endothelial progenitor cells and circulating endothelial cells, vascular endothelial growth factor, matrix metalloproteinase-9, C-reactive protein, and erythrocyte sedimentation rate were significantly higher in patients with Behcet’s disease. All of these parameters except circulating endothelial cells were also found to be higher in patients with active disease than in patients with inactive disease. Early endothelial progenitor cells showed significant correlations with C-reactive protein and circulating endothelial cells.Study limitations: The cross-sectional nature of the study and patient characteristics such as being under treatment, which can affect endothelial progenitor cells numbers.Conclusion: The increase in endothelial progenitor cells may play an essential role in the repair of endothelial injury in Behcet’s disease, especially in the active period of the disease. Thus, endothelial progenitor cells can indicate the disease activity. In addition, endothelial progenitor cells and circulating endothelial cells can be used as endothelial repair and injury markers for Behcet’s disease, respectively.     

Recombinant human erythropoietin stimulates vasculogenesis and wound healing in a patient with systemic sclerosis complicated by severe skin ulcers

Systemic sclerosis (SSc) is often complicated by severe skin ulcers that are unresponsive to traditional treatments. Vascular alterations are responsible for the ischaemic features of the disease in both the skin and visceral organs. Defective neoangiogenesis correlates with an abnormally reduced quantity of circulating endothelial progenitor cells (EPCs) caused by impaired maturation potential and proliferative capacity of bone-marrow endothelial stem cells. We report a patient with nonhealing cutaneous ulcers successfully treated with recombinant human erythropoietin (rHuEPO). The possible biological effects of this drug were also investigated. Before rHuEPO treatment, the bone-marrow sample contained reduced numbers of EPCs, which were functionally impaired. After a 6-month rHuEPO cycle, a marked increase in endothelial progenitor markers was seen, along with a significant reduction in their apoptotic rates. The clinical and laboratory data variations before and after rHuEPO treatment give new insights into the pathogenetic role of impaired endothelial stem-cell maturation and defective neoangiogenesis in patients with SSc.     

系统性红斑狼疮患者B淋巴细胞刺激因子及其受体BAFF-R的表达

目的 探讨系统性红斑狼疮（SLE）患者外周血单一核细胞（PBMC）中B淋巴细胞刺激因子（BLyS）及其受体BAFF—R（属于肿瘤坏死因子家族的B细胞活化因子受体）的表达情况。方法 采用半定量逆转录聚合酶链反应（RT-PCR）以及免疫印迹法（westem blot）分别检测28例SLE活动期患者及24例SLE稳定期患者PBMC中BLyS与BAFF—RmRNA及蛋白的表达水平，并以21例健康志愿者作对照；同时将两者mRNA及蛋白的表达水平与SLE疾病活动程度进行相关分析。结果 SLE活动期、稳定期患者PBMC中BLyS与BAFF—R mRNA及蛋白的表达水平均明显高于正常人（P〈0．01），活动期患者的表达高于稳定期患者（P〈0．01）；BLyS mRNA及蛋白的表达与狼疮活动指数（SLEDAI）呈显著正相关（P〈0．01），BAFF—R mRNA及蛋白的表达与SLEDAI无明显相关性（P〉0．05）。结论 BLyS及其受体BAFF—R可能参与了SLE的发病机制。     

SLE患者T淋巴细胞IL-13受体α1基因表达及其调节序列甲基化状态的研究

目的 研究SLE患者外周血T淋巴细胞IL-13受体α1（IL-13Rα1）基因mRNA的表达及IL-13Rα1基因调节序列甲基化状态。方法 免疫磁珠法（MACS）分离10例SLE患者和6例正常人外周血CD4+和CD8+ T细胞，采用实时荧光定量PCR检测T细胞中IL-13Rα1 mRNA的表达，并用甲基化特异性PCR（MSP）方法检测IL-13Rα1基因调节序列的甲基化水平。结果 活动期SLE患者CD4+ T细胞中IL-13Rα1 mRNA表达水平为2.224 ± 0.251，非活动期SLE患者为1.712 ± 0.132，正常人组为1.104 ± 0.044，三组间比较，差异均有统计学意义（P < 0.05）；CD8+ T细胞中IL-13Rα1 mRNA表达水平活动期、非活动期及正常人组分别为1.672 ± 0.142，1.410 ± 0.154，1.238 ± 0.106，活动期组与正常人组比较差异有统计学意义（P < 0.05），而非活动期组与正常人组、活动期组与非活动期组比较，差异均无统计学意义（P > 0.05）。CD4+ T细胞中IL-13Rα1基因甲基化指数活动期SLE患者为0.454 ± 0.023，非活动期为0.635 ± 0.065，正常人为0.844 ± 0.097，三组间比较，差异均有统计学意义（P < 0.05）；CD8+ T细胞中IL-13Rα1基因甲基化指数三组间比较，差异均无统计学意义（P > 0.05）。SLE患者外周血CD4+，CD8+ T细胞IL-13Rα1 mRNA的表达与疾病活动度（SLEDAI评分）呈正相关（r = 0.79，P < 0.01；r = 0.76，P < 0.05）；CD4+ T细胞的IL-13Rα1基因的甲基化水平与疾病活动度（SLEDAI评分）呈负相关（r = -0.89，P < 0.01）；CD4+ T细胞IL-13Rα1 mRNA表达与其调节序列的甲基化水平呈负相关（r = -0.84，P < 0.01）。结论 SLE的发生发展可能与DNA低甲基化导致SLE患者T细胞过度表达IL-13Rα1有关。