Expression and differential regulation of HLA-G isoforms in the retinal pigment epithelial cell line,ARPE-19

The purpose of this study was to examine if HLA-G is expressed in the retinal pigment epithelium (RPE) cells of the eye. The RPE comprises the outer most layer of the retina and as such defines the interface to the blood and contributes to the immune privilege in the posterior part of the eye. One way the RPE might be regulating the immune system could be by expressing the non-classical human leukocyte antigen (HLA) molecule, HLA-G. We therefore sought to define if the RPE cell line, ARPE-19, expressed HLA-G and analyse the regulation as a response to pro-inflammatory cytokines. This was done by digital droplet PCR, measuring the gene expression of HLA-G in total RNA. The protein expression was analysed by immunohistochemistry and by immunofluorescence followed by confocal microscopy and the expression of the HLA-G isoforms was explored by fragment analysis. In the current study, we show that HLA-G is expressed by ARPE-19 cells and is upregulated as a response to pro-inflammatory cytokines. Moreover, we are the first to describe a differential regulation of the HLA-G isoforms as a direct response to stimulation. These results might indicate that HLA-G can be part of the immune privilege of the posterior part of the eye, but further experiments on primary RPE cells are needed.     

骨髓间充质干细胞移植对脑缺血大鼠神经功能及IL-10、TNF-α水平的影响

目的:探讨骨髓间充质干细胞(MSC)移植治疗对大鼠脑缺血后神经功能及脑组织中IL-10、TNF-α表达的影响。方法:原代分离培养MSC。27只大鼠随机分为3组,各9只:假手术组,仅剪开皮肤;模型组,制作局灶性大脑中动脉闭塞(MCAO)模型,术后24 h尾静脉注射PBS;MSC组,制作MCAO模型,术后24 h尾静脉注射移植MSC。术后第3天行改良大鼠神经功能缺损评分(m NSS)评定大鼠神经功能;行为学测试后取梗死灶周围缺血半暗带脑组织,western blot法检测肿瘤坏死因子(TNF)-α、白细胞介素(IL)-10蛋白表达水平。结果:与模型组比较,MSC组m NSS评分升高(P0.05),脑组织蛋白IL-10表达水平上调(P0.05),TNF-α表达水平下调(P0.05)。结论:尾静脉移植MSC治疗可改善局灶性脑缺血大鼠神经功能,可能与移植MSC后脑组织中TNF-α和IL-10水平有关。     

醒脑静对兔心肌缺血再灌注损伤的抗炎性因子和改善凝血-纤溶功能障碍的作用

目的探讨醒脑静预处理对心肌缺血再灌注损伤（MIRI）的抗炎性因子的影响和改善凝血-纤溶功能障碍的作用。方法20只新西兰兔随机分成安慰剂组与醒脑静组，各10只。结扎冠状动脉造成MIRI模型，分别在冠状动脉结扎前、结扎后1h、再灌注后1h，取心房血分别检测肿瘤坏死因子-α（TNF-α）、组织型纤溶酶原激活物（t-PA）、纤溶酶原激活物抑制物-1（PAI-1）、肌酸激酶（CK）水平。同时观察各组梗死范围。结果冠状动脉结扎后醒脑静组TNF-α、PAI-1、CK水平较安慰剂组显著降低（P〈0．01），心肌梗死面积显著小于安慰剂组（P〈0．01）。结论醒脑静预处理对MIRI有良好的抗心肌缺血再灌注损伤作用。     

Differential nitric oxide and TNF-α production of murine Kupffer cell subfractions upon priming with IFN-γ and TNF-α

Abstract: Aims/Background: We have previously shown a striking heterogeneity of naive murine Kupffer cells (KC) that depends on cell size. Methods: In the present study, we demonstrate a shift in response of KC fractions separated on cell size by countercurrent elutriation upon priming with tumor necrosis factor-α (TNF-α) or interferon-γ (IFN-γ). Results: Whereas unprimed large KC are most active in the production of TNF-α and nitric oxide (NO), after priming of KC with TNF-α predominantly small and intermediate sized KC produce TNF-α in response to bacteria. Priming with IFN-γ enhanced NO production in all KC. A strong synergy, with respect to production of NO, was observed when KC subfractions were exposed to a combination of TNF-α and IFN-γ. Concerning TNF-α production, priming of KC subfractions seemed to induce a shift of activity from large KC to smaller KC. Conclusions: The present data demonstrate a clear heterogeneity among murine KC with respect to immunologic response to stimuli. These results demonstrate that KC have different functions in immunologic reactions that seem to be related to size.     

High prevalence of hidradenitis suppurativa symptoms in axial spondyloarthritis patients: A possible new extra-articular manifestation

Background

Spondyloarthritis (SpA), a chronic inflammatory, rheumatic disease, and hidradenitis suppurativa (HS), a chronic, debilitating, inflammatory skin disease, share several clinical and pathophysiological features, such as the association with inflammatory bowel disease and elevated cytokine levels IL-17 and TNF-α. Recently, SpA was reported to be more prevalent (2.3–28.2%) in patients with HS than in the general population. Conversely, the prevalence of HS in SpA is not exactly known.

西宁地区老年心力衰竭患者血浆脑钠肽和炎性因子水平变化的分析

目的探讨西宁地区(海拔2 260米以上)老年心力衰竭患者血浆脑钠肽(BNP)、C反应蛋白(CRP)、TNF-α水平变化的临床意义。方法将65例心力衰竭患者分为心衰组,同期选择34例健康体检者为对照组。采用美国博适Triage干式快速定量心力衰竭诊断仪检测血浆BNP水平、放射免疫法测定TNFα-水平、ELISA法测定CRP水平,同时行心脏超声检查。20例NYHAⅡ~Ⅳ级患者,常规抗心力衰竭治疗平均3个月后再次接受上述检查并留取血标本。结果(1)心衰组患者血浆BNP、CRP、TNF-α水平明显高于对照组,差异显著(P<0.01);(2)NYHAⅡ~Ⅳ级患者血浆BNP水平较治疗前明显下降;(3)心衰组患者血浆BNP水平在NYHA不同级别之间差异显著,与NYHA呈正相关(r=0.458,P<0.01),与LVEF呈负相关(F=-0.715,P<0.01)。结论血浆BNP、CRP、TNF-α参与了心力衰竭的发病过程,监测上述指标,可预测心力衰竭患者远期NYHA恢复情况,也可作为疗效观察及判定的重要参数。     

Plasma cytokines in graft vs host disease and complications following bone marrow transplantation

A cohort of 201 autologous or allogeneic bone marrow transplanted (BMT) patients were included for studying the evolution of circulating tumor necrosis factor alpha (TNF-alpha) and interleukin 6 (IL-6) by means of repeated plasma determinations. IL-6 levels were high during major transplant-related complications (TRC) or severe graft vs host disease (GVHD). High levels of TNF-alpha seemed to be associated with chronic GVHD but not with acute GVHD or TRC.     

急性心肌梗死患者早期干预过程中血浆P选择素水平的动态变化

目的观察急性心肌梗死(AMI)患者早期干预过程中血浆P选择素、TNF-α以及细胞间黏附分子-1(ICAM-1)水平的动态变化和早期再灌注治疗对其水平的影响。方法AMI患者31例(AMI组),经冠状动脉造影证实的稳定性心绞痛(SAP)患者17例(SAP组),冠状动脉造影正常者19例(对照组)。分别于发病6、12、24、487、2 h抽取静脉血。使用ELISA法测定血浆P选择素、TNF-α及ICAM-1水平。结果AMI组患者发病6 h内P选择素的浓度显著高于SAP组和对照组(P<0.01);12 h P选择素浓度进一步升高;24 h P选择素明显下降,但仍高于SAP组和对照组(P<0.05);48、72 h P选择素浓度与对照组比较无统计学差异。AMI组患者发病6 hTNF-α与ICAM-1即升高,24 h达高峰,72 h后仍高于正常。直线相关分析,AMI患者发病6 h的P选择素与TNF-α、ICAM-1水平呈正相关(r=0.76,P<0.01;r=0.69,P<0.01)。结论早期成功干预治疗使P选择素很快达峰值,然后迅速下降,但对TNF-α和ICAM-1水平变化的影响则不显著。     

H2O2、LPS 和 TNF-α诱导人类脐静脉内皮细胞损伤的实验研究

目的探讨人类脐静脉内皮细胞损伤模型建立方法,为体外研究血管内皮细胞提供实验基础。方法分离培养人类脐静脉内皮细胞,分别采用不同浓度的过氧化氢（H2O2）、脂多糖（LPS）、肿瘤坏死因子（TNF）‐α刺激细胞,孵育不同时间后,采用四甲基偶氮唑盐（MTT）法检测细胞活力（OD值）。结果各浓度H2O2损伤组较对照组OD值显著降低（P＜0．01）,不同浓度H2O2损伤组间OD值无统计学差异（P＞0．05）。0．1μg／mLLPS损伤组与对照组OD值无统计学差异（P＞0．05）;其余各浓度LPS损伤组较对照组OD值显著降低（P＜0．05）,且LPS浓度在一定范围内,OD值随LPS浓度的增加及作用时间的延长而降低,具有一定的浓度与时间依赖性。各浓度TNF‐α损伤组较对照组OD值显著降低（P＜0．01）,且TNF‐α浓度在一定范围内,OD值随TNF‐α浓度的增加及作用时间的延长而降低,具有一定的浓度与时间依赖性。结论H2O2、LPS和TNF‐α能体外损伤人类脐静脉内皮细胞,成功建立人类脐静脉内皮细胞损伤模型,且在一定浓度范围内各损伤因子对人类脐静脉内皮细胞损伤程度呈浓度和时间依赖性。     

子痫前期孕妇血清细胞因子信号转导负调控因子-3、肿瘤坏死因子-α及白细胞介素-10水平变化的临床价值

目的 研究子痫前期孕妇血清细胞因子信号转导负调控因子(SOCS)3、肿瘤坏死因子(TNF)-α及白细胞介素(IL)-10水平及临床价值.方法 将2012年1月至2013年12月在上海交通大学医学院附属国际和平妇幼保健院确诊的40例子痫前期孕妇纳入研究的观察组,同期在上海交通大学医学院附属国际和平妇幼保健院分娩的无妊娠并发症的40例孕妇纳入研究的对照组,检测血清中SOCS3、TNF-α、IL-10的含量并分析其相关性.结果 (1)观察组与对照组孕妇血清中SOCS3的mRNA含量分别为38 ±6、100±16;IL-10的mRNA含量分别为41±7、100±15;TNF-α的mRNA含量分别为226±40、100±19;(2)观察组孕妇血清中SOCS3、IL-10的蛋白含量低于对照组孕妇(P＜0.05),TNF-α的蛋白含量高于对照组孕妇(P ＜0.05);(3) SOCS3的mRNA含量和蛋白含量分别与TNF-α的mRNA含量和蛋白含量呈负相关,分别与IL-10的mRNA含量和蛋白含量呈正相关.结论 子痫前期孕妇血清中SOCS3含量显著降低,进而导致抑制Th1型细胞因子TNF-α表达和促进Th2型细胞因子IL-10表达的作用减弱,在疾病的发生过程中发挥重要作用.