ICAM-1及其受体在高致病性禽流感病毒性肺炎中的作用研究

本文应用高致病性禽流感病毒性肺炎(Highly pathogenic avianinfluenza viral pneumonia,HPAIVP)小鼠模型,采用免疫组织化学染色方法,对实验组和对照组小鼠肺组织中ICAM-1及其受体的表达含量进行检测.研究发现实验组小鼠肺组织中ICAM-1及其受体表达含量在攻毒后1d即开始升高,第4天达高峰.就攻毒剂量来说,以100LD50/50 μL攻毒组小鼠肺组织中ICAM-1及其受体阳性表达量升高最明显.并通过ELASA方法进一步证实了这种变化.结果表明ICAM-1及其受体在HPAIVP中呈现高表达,可能在其发病中发挥了重要作用.     

减蛋综合征病毒在不同品系小鼠体内的组织分布

目的通过人工感染减蛋综合征病毒(egg drop syndrome virus,EDSV),观察病毒在不同品系小鼠体内增殖情况以及动态变化规律,为EDSV构建载体提供理论依据与数据支持。方法选取免疫系统正常的BALB/c小鼠、T细胞免疫缺陷裸鼠(Nu)以及高度免疫缺陷小鼠(NSG)为研究对象,每品系32只,雌性,5～6周龄,经腹腔注射人工感染EDSV,分别于攻毒后1、3、5、7、14、21、28、35 d采集血清,应用间接ELISA方法进行抗体监测;选择攻毒后1、7、14、21、28 d小鼠,采集心脏、肺、肝、脾、肾、小肠、子宫、气管、食管、脑10种组织,应用荧光定量PCR相对定量比较Ct法(△△CT)进行各组织内病毒载量的检测。结果 BALB/c小鼠于攻毒后3 d即可在血清内检测到抗体的表达,14 d抗体水平达到最高,并一直维持至监测期内35 d;Nu小鼠也可于攻毒后3 d检测到抗体,表达水平较BALB/c小鼠有所降低,攻毒14 d后,Nu小鼠血清中抗体水平出现下降,至35 d抗体一直维持在较低的水平;NSG小鼠在整个监测过程中,抗体水平一直处于阴性状态。核酸相对定量结果显示,BALB/c小鼠感染后1 d,肝组织中的病毒表达量最高,达到5.45个数量级,其次由高到低依次是脾、食管、子宫、小肠、肺、气管、肾、心脏,脑组织中病毒含量最低,随感染时间的延长,各组织内病毒表达量较感染1 d均有所下降,至攻毒后28 d,肝、脾病毒表达量依然维持着较高的水平;Nu小鼠和NSG小鼠感染1 d表现为脾中病毒表达量最高,分别为3.95和4.05个数量级,其次为肝,攻毒28 d,两种小鼠体内各器官内仍可以检出阳性信号,肝、脾病毒表达量较高。结论 EDSV可刺激小鼠产生免疫应答,在免疫缺陷小鼠体内抗体水平表达量较低。该病毒在小鼠体内有肝、脾等组织嗜性,为EDSV开发成为载体以及在实验动物模型上的进一步研究与应用提供了参考数据。     

红霉素对慢性阻塞性肺疾病大鼠ICAM-1、MMP-9的表达影响

目的:观察慢性阻塞性肺疾病(COPD)大鼠肺组织中ICAM-1及MMP-9的表达及红霉素的干预作用。方法:复制COPD大鼠模型,并用红霉素干预,收集支气管肺泡灌洗液行细胞学计数和分类检查;采用HE染色观察病理形态变化;免疫组化法检测大鼠支气管肺组织ICAM-1、MMP-9的表达。结果:与模型组比较,干预组支气管肺组织中ICAM-1、MMP-9表达显著降低;模型组中ICAM-1、MMP-9的表达与BALF中白细胞总数及中性粒细胞数成正相关;ICAM-1与MMP-9的表达成正相关。结论:COPD大鼠肺组织中的ICAM-1、MMP-9表达明显升高,可能与COPD的发病机制有关;红霉素可降低ICAM-1、MMP-9的表达,可能是红霉素在COPD中抗炎症反应的作用机制之一。     

甲型H1N1流感病毒感染小鼠的发病机制

目的甲型H1N1流感病毒A/California/7/2009感染BALB/c小鼠,研究甲型H1N1流感病毒病毒性肺炎发病机制。方法 4～6周龄雌性BALB/c小鼠60只,随机分为2组,实验组和对照组,每组30只。CA7流感病毒滴鼻制备甲流病毒感染小鼠模型。攻毒后第5天解剖实验和对照组小鼠,取肺组织,测定肺组织中IL-2,IL-6,TNF-α含量。结果结果实验组肺组织中IL-6,TNF-α,水平明显高于对照组,IL-2水平明显低于对照组,差异均有显著性（P〈0.05）。结论 IL-6、TNF-α、IL-2这3种细胞因子在感染甲流病毒后的显著性变化与病毒感染后的肺组织病理损伤有密切的关系。     

小鼠不同组织及缺氧损伤后的大鼠心肌细胞中RIP3的表达

目的:检测小鼠组织中受体相互作用丝氨酸/苏氨酸蛋白激酶家族(RIPs)表达谱,并检测RIP3在大鼠心肌细胞缺氧损伤后的表达。方法:①采用荧光实时定量PCR分别检测RIPs家族基因在小鼠组织(心、肝、肺、肾、脑、小肠、骨骼肌、脾和主动脉)中的mRNA表达谱,并采用Western blot进一步检测RIP3在小鼠组织的蛋白表达谱。②将培养的大鼠心肌细胞分为缺氧组和对照组,缺氧组置于缺氧环境中培养48 h,采用western blot检测其中RIP3的表达变化。结果:①mRNA水平:RIP1 mRNA在脑组织中表达最高,心脏、肺、肾、骨骼肌较低;RIP2在心脏和肺表达量较其他组织高;RIP3在肠中表达较其他组织高出4倍以上,脑组织中未检测到RIP3表达;RIP4的表达以肺最高,而骨骼肌、脑和血管中表达量低。②蛋白水平:在小鼠组织中,RIP3表达以脑、骨骼肌中最高,心脏、肝、肺中表达较低。③培养的大鼠心肌细胞中,缺氧组心肌细胞的RIP3表达量显著高于对照组(P0.05)。结论:RIPs在小鼠组织中呈现差异表达,而在培养的大鼠心肌细胞缺氧损伤后RIP3表达升高。     

姜黄素预处理对沙漠干热环境热射病大鼠肺损伤及HMGB-1和ICAM-1 mRNA的影响

目的:研究姜黄素预处理对干热环境热射病大鼠肺损伤的保护作用及可能机制。方法:将50只SD大鼠随机分为5组(n=10):常温对照组(NC)、干热对照组(DHC)、低剂量姜黄素预处理组(50 mg/kg),中剂量姜黄素预处理组(100 mg/kg)及高剂量姜黄素预处理组(200 mg/kg)。NC、DHC组给予生理盐水灌胃,姜黄素预处理组给予不同剂量的姜黄素灌胃,每天1次,连续7天。第8天除NC组外,其余4组大鼠转移至西北特殊环境人工实验舱内进行实验,环境温度(41±0.5)℃,湿度(10±1)%。实验的第150分钟达到热射病状态,麻醉后取材。大鼠肺组织通过HE染色并进行肺损伤病理学评分,并应用RT-PCR检测肺组织HMGB1 mRNA和ICAM-1 mRNA的表达。结果:干热对照组大鼠肺组织病理评分、肺组织HMGB1和ICAM-1 mRNA表达较常温对照组、中、高剂量姜黄素预处理组均显著升高(P0.01),高剂量姜黄素预处理组大鼠肺组织病理评分明显低于低、中剂量姜黄素预处理组(P0.01),肺组织HMGB1和ICAM-1 mRNA表达在高剂量姜黄素预处理组明显低于低剂量姜黄素预处理组(P0.01)。肺组织HMGB1和ICAM-1 mRNA的表达均与肺损伤评分具有显著正相关性(r=0.629、0.689,P0.01),HMGB1 mRNA和ICAM-1 mRNA之间的表达也呈显著正相关性(r=0.437,P0.01)。结论:姜黄素可能部分通过抑制HMGB1表达的上调,减少下游的ICAM-1的表达来减轻炎症反应,从而发挥肺损伤保护作用。     

COPD大鼠肺组织中小窝蛋白-1 的表达及其与气道重塑的关系

目的:探讨小窝蛋白-1(Caveolin-1)在COPD大鼠肺组织中的表达及其与气道重塑的关系。方法:采用10周龄Wister大鼠20只,随机分为对照组与实验组,每组10只,对照组常规饲养,实验组采用烟熏法+内毒素法建立COPD模型。第91天处死全部大鼠,测定肺功能后取肺组织,采用免疫组织化学法检测Caveolin-1的表达;酶联免疫吸附试验(ELISA)检测各组大鼠肺组织匀浆中白细胞介素-8(IL-8)、肿瘤坏死因子-α(TNF-α)、转化生长因子-β1(TGF-β1)的含量;分析Caveolin-1的表达与IL-8、TNF-α、TGF-β表达的相关性。结果:与对照组比较,实验组大鼠的肺顺应性明显下降,肺弹性阻力及气道阻力显著升高,肺组织TNF-α、IL-8、TGF-β1的含量明显升高,Caveolin-1的表达明显降低,差异均有统计学意义(P0.05)。大鼠肺组织中Caveolin-1的表达与IL-8、TNF-α、TGF-β1含量呈显著负相关(P0.05)。结论:Caveolin-1在COPD大鼠肺组织中的表达明显降低,可能通过促进炎性细胞因子的释放与气道重塑参与COPD的发生和发展。     

肠淋巴再灌注加重肠系膜上动脉闭塞性休克大鼠肺部炎症反应

目的:研究肠系膜淋巴再灌注对肠系膜上动脉闭塞性(SMAO)休克大鼠肺部炎症反应的影响。方法:24只Wistar雄性大鼠均分为4组:SMAO组,MLR组,SMAO+MLR组,SHAM组。再灌注2h后,迅速留取肺组织,一部分制备组织匀浆,检测细胞间粘附分子(ICAM-1)和晚期糖基化产物受体(RAGE)。再另外选取固定位置肺部组织放入中性甲醛中固定,用于测定肺内HMGB1、RAGE的表达。结果:SMAO与SMAO+MLR组肺部组织匀浆ICAM-1、RAGE含量显著高于MLR与SHAM组,且SMAO+MLR组肺组织匀浆的ICAM-1、RAGE含量高于SMAO组。肺部组织内HMGB1和RAGE在MLR组与SHAM组基本不表达,或少量表达,MLR加重了SMAO休克模型中HMGB1和RAGE的表达。结论:MLR加重SMAO休克大鼠肺部炎症反应,进一步证实肠淋巴途径在SMAO休克发病学中具有重要作用,同时证实HMGB1及RAGE在SMAO休克大鼠的炎症失常反应中起重要作用。     

红霉素对慢性阻塞性肺疾病大鼠ICAM-1、MMP-9的表达影响

目的：观察慢性阻塞性肺疾病（COPD）大鼠肺组织中ICAM-1及MMP-9的表达及红霉素的干预作用。方法：复制COPD大鼠模型,并用红霉素干预,收集支气管肺泡灌洗液行细胞学计数和分类检查;采用HE染色观察病理形态变化;免疫组化法检测大鼠支气管肺组织ICAM-1、MMP-9的表达。结果：与模型组比较,干预组支气管肺组织中ICAM-1、MMP-9表达显著降低;模型组中ICAM-1、MMP-9的表达与BALF中白细胞总数及中性粒细胞数成正相关;ICAM-1与MMP-9的表达成正相关。结论：COPD大鼠肺组织中的ICAM-1、MMP-9表达明显升高,可能与COPD的发病机制有关;红霉素可降低ICAM-1、MMP-9的表达,可能是红霉素在COPD中抗炎症反应的作用机制之一。     

鸡产蛋下降综合征病毒NE4毒株对KM小鼠感染特性观察

目的初步探究鸡产蛋下降综合征病毒NE4（EDSV NE4）毒株对昆明小鼠的感染特性。方法用106TCID50攻毒量的EDSV NE4株对4～6周龄的KM小鼠进行攻毒试验,同时以正常尿囊液接种作为阴性对照。用荧光定量PCR对小鼠组织及粪便中的EDSV进行检测,同时用HE染色及免疫组化法对小鼠组织切片进行病理组织学观察和抗原定位。结果 EDSV攻毒对小鼠的采食情况产生一定影响,但并未引起明显的临床症状;攻毒组小鼠可产生针对EDSV特异性的抗体,HI抗体滴度可高达212,阴性对照组小鼠体内抗体检测为阴性;攻毒后小鼠大部分组织器官如肝脏、子宫、肾脏、肺脏等与攻毒7 d后粪便中均可检测到EDSV,阴性对照小鼠所有受检组织中均未检测到病毒;EDSV攻毒未能引起小鼠组织的病理学变化,攻毒后不同时间内,可在子宫、肺脏、肝脏、肾脏中可检测到阳性信号,最为典型的定殖位置是子宫腺体上皮细胞及肌层细胞的胞质中。结论 EDSV NE4株可以感染KM小鼠。     

Regulation of cellular adhesion molecule expression in murine oocytes,peri-implantation and post-implantation embryos

Expression of the adhesion molecules, ICAM-1, VCAM-1, NCAM, CD44, CD49d (VLA-4, α chain),and CD11a (LFA-1, α chain) on mouse oocytes, and pre- and peri-implantation stage embryos was exam-ined by quantitative indirect immunofluorescence microscopy. ICAM-1 was most strongly expressed at theoocyte stage, gradually declining almost to undetectable levels by the expanded blastocyst stage. NCAM,also expressed maximally on the oocyte, declined to undetectable levels beyond the morula stage. On theother hand, CD44 declined from highest expression at the oocyte stage to show a second maximum at thecompacted 8-cell/morula. This molecule exhibited high expression around contact areas between trophecto-derm and zona pellucida during blastocyst hatching. CD49d was highly expressed in the oocyte, remainedsignificantly expressed throughout and after blastocyst hatching was expressed on the polar trophecto-derm. Like CD44, CD49d declined to undetectable levels at the blastocyst outgrowth stage. Expression ofboth VCAM-1 and CD11a was undetectable throughout. The diametrical temporal expression pattern ofICAM-1 and NCAM compared to CD44 and CD49d suggest that dynamic changes in expression of adhesionmolecules may be important for interaction of the embryo with the maternal cellular environment as wellas for continuing development and survival of the early embryo.     

Regulation of cellular adhesion molecule expression in murine oocytes, peri-implant ation and post-implantation embryos

Expression of the adhesion molecules, ICAM-1, VCAM-1, NCAM, CD44, CD49d (VLA-4, a chain), and CDlla (LFA-1, a chain) on mouse oocytes, and pre- and peri-implantation stage embryos was examined by quantitative indirect immunofluorescence microscopy. ICAM-1 was most strongly expressed at the oocyte stage, gradually declining almost to undetectable levels by the expanded blastocyst stage. NCAM, also expressed maximally on the oocyte, declined to undetectable levels beyond the morula stage. On the other hand, CD44 declined from highest expression at the oocyte stage to show a second maximum at the compacted 8-cell/morula. This molecule exhibited high expression around contact areas between trophecto-derm and zona pellucida during blastocyst hatching. CD49d was highly expressed in the oocyte, remained significantly expressed throughout and after blastocyst hatching was expressed on the polar trophecto-derm. Like CD44, CD49d declined to undetectable levels at the blastocyst outgrowth stage. Expression of both     

模拟常压失事潜艇环境人血细胞表面CD55、CD59分布的变化

目的：探讨模拟固壳未破损失事潜艇环境条件暴露对人体血液免疫功能的影响及其变化的规律。方法：使用500m饱和潜水居住舱系统模拟失事潜艇固壳未破损舱室环境条件,控制舱内温度、PCO2、PO2,于进舱前、暴露第2、4、8d采晨空腹血,用流式细胞分析术测定红细胞、淋巴细胞、中性粒细胞表面CD55和CD59的分布变化。结果：红细胞膜表面CD55的分布在8d有显著升高（P〈0．01）;淋巴细胞膜表面CD55的分布在暴露的初、中期显著下降（P〈O．01）,后期明显恢复,与温度、PO2呈正相关（P〈O．01）,与PCO2呈负相关（P〈O．01）,CD59分布的变化与CD55变化类似。结论：氧分压和环境温度过低、二氧化碳分压过高都是诱发免疫活性细胞激活的有效因素。此环境暴露对血细胞自身保护作用有随时间累积的效应。     

中药复方连黄对小鼠T淋巴细胞亚群CD4~+、CD8~+的影响

目的探讨中药复方连黄对小鼠T淋巴细胞亚群CD4+、CD8+的影响.方法 T淋巴细胞亚群测定采用单克降抗体直接免疫荧光技术,通过流式细胞仪测定.结果经统计学分析,与对照组比较,中药复方连黄能不同程度地使CD4+、CD4+/CD8+升高,而使CD8+下降.结果表明,中药复方连黄对细胞免疫功能具有调节作用.     

小鼠肺脏间质树突状细胞形态学分布与观察

目的认识小鼠肺脏间质树突状细胞(dendritic cells,DC)的形态、数量和分布特点。方法运用光镜、电镜观察与免疫组化标记(CD11c、CD205、CD80、CD86及MHC-II类分子I-Ab)方法,观察研究C57BL/6小鼠肺脏间质树突状细胞的形态、数量和分布。结果小鼠肺脏间质树突状细胞具有与免疫器官及外周血中DC相同的超微形态特征;CD11c阳性DC含量约占肺脏总细胞数的8.5‰,广泛分布于肺间质中;表达CD205、CD80、CD86及I-Ab的成熟DC含量稀少,多位于间质血管周围。结论肺脏间质树突状细胞是一种脏器免疫前哨细胞,是肺脏免疫微环境的重要组成部分。     

Infection with respiratory syncytial virus enhances expression of native receptors for non-pilate Neisseria meningitidis on HEp-2 cells

Respiratory virus infections have been suggested to be predisposing factors for meningococcal disease. Respiratory syncytial virus (RSV) affects young children in the age range at greatest risk of disease caused by Neisseria meningitidis. It has been previously shown that glycoprotein G expressed on the surface of RSV-infected HEp-2 cells (a human epithelial cell line) contributed to higher levels of binding of meningococci compared with uninfected cells. The aim of the present study was to examine the effect of RSV infection on expression of surface molecules native to HEp-2 cells and their role in bacterial binding. Flow cytometry and fluorescence microscopy were used to assess bacterial binding and expression of host cell antigens. Some molecules analysed in this study have not been reported previously on epithelial cells. RSV infection significantly enhanced the expression of CD15 (P < 0.05), CD14 (P < 0.001) and CD18 (P < 0.01), and the latter two contributed to increased binding of meningococci to cells but not the Gram-positive Streptococcus pneumoniae.     

The role of CD40-CD154 interaction in cell immunoregulation

CD40, a member of the nerve growth factor/tumor necrosis factor receptor superfamily, and its ligand, CD154, play essential roles in cell immune responses. The results of many studies have indicated that CD40-CD154 interaction can upregulate costimulatory molecules, activate antigen-presenting cells (APCs), influence T-cell priming and T-cell-mediated effector functions as well as participate in the pathogenic processing of chronic inflammatory diseases, such as autoimmune diabetes, graft rejection, atherosclerosis, and cancer. Ligation of CD40 on cancer cells was also found to produce a direct growth-inhibitory effect through cell cycle blockage and/or apoptosis with no overt side effects on normal cells and treatment with CD154 can heighten tumor rejection immune response as well. However, systemic treatment with CD154 has some potential risks. Therefore, searching for efficient and safe strategies of CD154-based cancer therapy has been a hot topic in human cancer research. This review focuses on the latest discovered functions of CD40-CD154 interaction in cell immune responses and on the new findings of CD154-based human cancer therapy.     

Leukocyte trafficking in free-flowing cerebrospinal fluid of normal rhesus macaques (Macaca mulatta)

Abstract: Cellular components in free-flowing cerebrospinal fluid (CSF) of normal rhesus macaques were characterized. Microscopic counting enumerated the total number of leukocytes, percentage of polymorphonuclear cells (PMN), leukocytes with nonspecific esterase (NSE), and those reducing nitro blue tetrazolium (NBT). Flow cytometric analysis further identified CD4, CD8, CD14, and CD20 positive leukocytes. These experiments established reliable techniques for evaluating cellular components in CSF from rhesus macaques and documented the difference in the CD4/CD8 ratio between peripheral blood (PB) and CSF compartments under normal physiological conditions.     

Programmed cell death: the influence of CD40, CD95 (Fas or Apo-I) and their ligands

Programmed cell death (PCD) or apoptosis is a process whereby developmental or environmental stimuli activate a specific series of events that culminate in cell death. PCD is essential for normal development and abnormality in the process can lead to defects ranging from embryonic lethality and tissue-specific perturbation of postnatal development to a high susceptibility to malignancy. Therapeutics that modulate the regulation of PCD may provide a new opportunity for the treatment of the PCD related diseases and cancer. CD40 and CD95 (Fas/Apo-I) are transmembrane proteins of the nerve growth factor/tumour necrosis factor α receptor superfamily. The death signal of PCD occurs when the CD95 receptor on the cell surface binds to the CD95 ligand (CD95L) or to the anti-CD95 monoclonal antibody (mAb). In contrast, PCD could be inhibited by the survival signal mediated from the binding of the CD40 receptor to the CD40 ligand (CD40L) or to the anti-CD40 mAb. In this review, the interaction of CD40/CD40L and CD95/CD95L on PCD in normal and malignant cells is discussed.     

低血容量休克复合内毒素血症时血和组织髓过氧化物酶的变化

研究观察失血性休克复合内毒素血症时血和组织髓过氧化物酶的变化规律。将雄性wistar大白鼠随机分为对照组、缺血组、缺血再灌流组和缺血再灌流复合内毒素组。用改良的髓过氧化物酶 (MPO)测定方法 ,测定血、肺和小肠组织MPO及相关指标的变化。结果显示肺组织MPO活性从失血性休克末开始升高 ,致内毒素血症时出现峰值 ;小肠组织MPO的活性在失血再灌流后显著升高 ,但在失血性休克复合内毒素血症后显著降低 ;血MPO活性于失血性休克和失血再灌流后均无显著性变化 ,复合内毒素后显著降低。结果表明失血再灌流后肺组织PMN扣留、聚集显著增加 ,内毒素血症促进PMN在肺中的扣留 ,这些变化与PMN上CD11b和CD18表达上调有关 ,提示失血再灌注复合内毒素时组织细胞损伤与PMN的粘附、扣留、激活有关。