Fractalkine对类风湿关节炎患者成纤维样滑膜细胞中NF-κB活化及内源性fractalkine mRNA表达的影响

目的:观察Fractalkine(FKN)对类风湿关节炎(RA)患者成纤维样滑膜细胞(FLS)核转录因子κB (NF-κB)活化以及内源性FKN mRNA表达的影响.方法:通过组织块法培养RA-FLS.在RA-FLS中加入100 μg/L FKN,分别作用0 h、1 h及2 h,用Western blotting检测RA-FLS胞浆及胞核中NF-κB p65蛋白表达量变化以明确NF-κB的活化情况;加入100 μg/L重组人FKN分别作用0 h、12 h、18 h后,用RT-PCR检测FKN mRNA表达的变化.结果:重组人FKN刺激1 h后,RA-FLS胞浆中NF-κB p65蛋白水平明显低于无FKN刺激的对照组(P<0.05);FKN刺激后2 h,细胞核中NF-κB p65蛋白水平较对照组明显升高(P<0.05).100 μg/L重组人FKN刺激RA-FLS,呈时间依赖方式诱导内源性FKN mRNA表达.FKN刺激RA-FLS18 h,细胞中FKN mRNA的表达水平明显升高(P<0.05).结论:外源FKN可刺激RA-FLS内源性的FKN mRNA表达上升,提示RA-FLS中可能存在FKN的正反馈现象.FKN对NF-κB有活化作用,可能对RA患者关节中炎症的启动、血管生成和骨质破坏有重要作用.     

氧烛对高原人体血氧饱和度及血乳酸的影响

目的观察在高原利用氧烛建立富氧室对移居高原男青年血氧饱和度（SaO2）及血乳酸（BLA）的影响。方法 8名进驻高原1年的受试者在海拔3700 m夜间睡眠,监测其常氧和富氧（氧浓度为24%～25%）条件下的SaO2变化;起床脱离富氧室3 h后空腹采集肘静脉血,检测BLA在富氧前、富氧后的变化。结果富氧较常氧下受试者SaO2增高（P〈0.01）,富氧组BLA与常氧组比较降低（P〈0.01）。结论用氧烛制作富氧室可显著改善人体高原缺氧状态。     

壳聚糖微球转染人IL-1Ra与TGF-β1基因治疗兔骨关节炎

目的 探讨壳聚糖微球转染人IL-1Ra与TGF-β1基因治疗兔膝关节早期骨关节炎的方法与效果.方法 制备分别包裹IL-1Ra质粒DNA和TGF-β1质粒DNA的壳聚糖微球缓释系统,分组向兔膝关节早期骨关节炎模型关节腔注射含IL-1Ra基因和(或)TGF-β1基因的壳聚糖微球、不含上述基因的壳聚糖溶液,定期处死,使用ELISA检测关节腔灌洗液的IL-1Ra、TGF-β1浓度,取关节标本Mankin评分、苏木精-伊红(HE)染色、番红O染色及免疫组化检测.结果 经过60 d观察,壳聚糖微球转染基因组的IL-1Ra与TGF-β1的基因可持续表达,双基因组的软骨标本从外观、染色观察,损伤程度轻于单基因组.双基因组的Mankin评分明显低于单基因组(P＜0.05),单基因组的Mankin评分明显低于空白组(P＜0.05).结论 关节腔注射壳聚糖微球转染IL-1Ra与TGF-β1基因能抑制软骨的退变和促进软骨的修复.

Abstract：

Objective To explore the method and effect of transinfection of rabbit early knee osteoarthritis models via chitosan microsphere with gene of recombined human IL-1Ra gene and TGF-β1 gene. Methods Chitosan microspheres with plasmids of IL-1Ra gene and TGF-β1 gene, and rabbit early knee osteoarthritis models were prepared. Rabbits in different groups had intra-articular injections of chitosan microsphere containing IL-1Ra gene and / or TGF-β1 gene, and chitosan solution as control group before being executed regularly and randomly. The joint specimens were evaluated by HE staining, lycopene red O staining and immunohistochemical analysis and Mankin's score. ELISA was used for detection of IL-IRa and TGF-β1 concentration of articular cavity fluid in each group. Results The control group was consistent with the pathological changes of early OA. In co-transinfection group, judging from the appearance and staining of cartilage,the OA damage of the specimens was less serious than other groups'. Its Mankin's score was significantly lower than single-gene transinfection group (P ＜ 0.05), and the latters Mankin's score were significantly lower than control group (P ＜ 0.05). Conclusion Intra-articular injection of chitosan microspheres containing both IL-1Ra gene and TGF-β1 gene could inhibit the degeneration of cartilage and promote cartilage repair.     

透明质酸修饰壳聚糖/pDNA纳米微球的制备及体外转染软骨细胞

目的 以透明质酸(HA)修饰基因载体壳聚糖(CS)纳米微球,制作一种新型的HA/CS/pDNA基因转染系统,观察其结构特征及体外对软骨细胞的转染能力.方法 将不同比例的HA和CS与负载增强型绿色荧光蛋白基因(EGFP)的质粒DNA(pDNA)以复凝聚法制成纳米微球,以扫描电镜检测纳米微球形态,Zeta电位粒度分析仪测定其粒径、表面电位;凝胶电泳阻滞实验观察CS和pDNA的结合力;体外转染兔关节软骨细胞,以流式细胞仪及荧光显微镜检测转染效率.结果 HA/CS/pDNA纳米微球多呈球形,粒径为(223±51)nm,表面Zeta电位为(17.4±6.1)mV,可有效保护pDNA免受 DNaseⅠ的降解.体外转染实验证明HA/CS/pDNA纳米微球能介导pEGFP转染软骨细胞并在细胞内表达绿色荧光蛋白,转染率达(15.450±0.404)%,比裸pDNA组和CS/pDNA组有更高的转染效率(P<0.05).结论 复凝聚法制备的HA/CS/pDNA纳米微球是一种有效的新型非病毒基因转染系统,对软骨细胞有着潜在的靶向基因转染能力.

Abstract：

Objective To prepare hyaluronic acid (HA)-modified chitosan (CS)/pDNA (HA/CS/pDNA) nanoparticles as novel gene vectors, study their structural characteristics and gene transfection efficiency for chondrocytes in vitro in rabbits. Methods The HA/CS/pDNA nanoparticles were prepared by a DNA (pDNA), which load enhanced green fluorescent protein (EGFP) gene. The morphology of the nanoparticles was observed under the transmission electron microscopy. The sizes and zeta-potentials of the nanoparticles were measured by a Marven-nano laser diffractometer. The binding of pDNA was evaluated by agarose gel electrophoresis analysis. The gene transfection experiments in vitro were performed with the chondrocytes of rabbits. The gene transfection efficiency was measured by using flow cytometry and under the fluorescence microscopy. Results HA/CS/pDNA nanoparticles were mainly spherical, with an average size of (223±51) nm, and zeta-potential of (17.4±6.1) mV. The agarose gel electrophoresis analysis confirmed that they could effectively protect pDNA from degradation against DNase I. Gene transfection in vitro proved that HA/CS/pDNA nanoparticles could be efficiently transfected into chondrocytes of rabbits, the expression of green fluorescent proteins was observed under the fluorescent microscopy, and the transfection efficiency reached as high as (15.450±0.404)%, significantly higher than that of the naked pDNA or the CS/pDNA nanoparticles (P<0.05). Conclusion HA/CS/pDNA nanoparticles were an effective novel non-viral gene transfer vector, which possessed the potential targeting transfection ability on chondrocytes.     

急性心肌梗死伴2型糖尿病患者内皮祖细胞动员障碍

目的观察急性心肌梗死伴2型糖尿病患者血浆缺血相关因子血管内皮生长因子和基质细胞衍生因子水平,骨髓内皮祖细胞动员是否存在障碍,以及基质细胞衍生因子、血管内皮生长因子-内皮祖细胞动员通路是否存在异常。方法采用密度梯度离心法分离外周血单个核细胞,用流式细胞仪检测急性心肌梗死后不同时间点(1、3、5、7、14和28天)外周血CD45-/low /CD34 /CD133 /KDR 早期内皮祖细胞数量。酶联免疫吸附法检测血浆中血管内皮生长因子、基质细胞衍生因子以及高敏C反应蛋白的浓度。结果急性心肌梗死伴2型糖尿病患者外周血内皮祖细胞动员高峰(第7天)较急性心肌梗死非糖尿病患者(第5天)延迟且显著减少[(140±48)/106比(246±100)/106,P<0.05]。糖尿病组血浆血管内皮生长因子(第5天:277±95ng/L比168±35ng/L,P<0.05)、基质细胞衍生因子(第5天:3835±402ng/L比3287±384ng/L,P<0.05)以及高敏C反应蛋白(第3天:55.55±14.88mg/L比36.92±14.83mg/L,P<0.05)在高峰点的水平显著高于非糖尿病组。结论糖尿病患者心肌梗死后组织缺血程度较重,但组织缺血后基质细胞衍生因子、血管内皮生长因子-内皮祖细胞动员通路存在障碍,这可能是糖尿病患者缺血后血管新生功能障碍,发生急性心肌梗死后预后较差的原因之一。     

QTc、JTc间期与QT/TQ比的正常值及临床意义

本文报道1 309名正常人的QTc间期(<440ms),JTc间期(<360ms)和QT/TQ比(<1.1),并与急性心肌梗塞(AMI)和单纯完全性左、右束支传导阻滞(CLBBB、CRBBB)患者对比,发现AMI、CLBBB、CRBBB患者QTc和QT/TQ比均明显大于正常组(P<0.01),而JTc间期仅AMI组显著长于正常组(P<0.01)。结果提示,JTc间期较QFc间期能更好地反映心室肌复极状态,对心室除极顺序正常和异常均适用。若能综合分析以上三值,对临床指导意义更大。     

水银血压计在眼眶静脉畸形患者检查中的应用

<正>在临床工作中,眼眶静脉畸形的患者需进行各项诊断检查,此类患者在行超声波检查、彩色多普勒超声(color doppler imaging,CDI)检查或行CT扫描时必须进行颈部加压(压迫双侧颈静脉),根据加压前后眼眶内血供变化的对比,对眼眶静脉畸形患者做出进一步诊断。临床上常采取手压双侧颈静脉、俯卧低头位、颈部缠绕止血带进行颈部加压,这些方法常     

慢性咽炎患者需要服用抗生素吗?

有的人得了慢性咽炎之后,认为既然是咽部发炎,就应该用消炎药,于是自作主张,口服红霉素、螺旋霉素、麦迪霉素、头孢菌素、复方磺胺甲嗯唑片(复方新诺明)等抗菌药物,或者注射青霉素,甚至长期使用上述药物,结果大多疗效不明显或根本无效。这是什么原因呢?