双氯芬酸钠对大鼠骨髓间充质干细胞影响的浓度、时间效应

目的通过不同浓度的双氯芬酸钠干预体外培养的大鼠骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs),观察其对大鼠骨髓间充质干细胞增殖、凋亡及成骨分化能力的影响;不同药物干预时间对细胞成骨分化能力的影响。方法不同浓度的双氯芬酸钠(6.4、3.2、1.6、0.8、0.4mg/L)干预第4代(P4)BMSCs,通过MTS法检测细胞增殖活性;AnnexinⅤ-FITC法检测细胞凋亡;根据茜素红染色钙结节的数量,初步检测细胞成骨能力。并选取1.6mg/L的药物浓度,不同时间点和时长进行干预,检测细胞成骨分化能力的变化。结果随着双氯芬酸钠浓度的升高,BMSCs细胞增殖活性逐渐降低,细胞凋亡比率升高,钙结节数量减少。药物在第1~14天期间干预能够减少钙结节的形成,在第14~21天期间干预对钙结节的形成无明显影响。结论双氯芬酸钠可影响BMSCs的增殖、凋亡及成骨分化能力,且呈浓度和时间依赖性,短期的作用效果是部分可逆的。     

Huntington Disease in Asia

Objective:

The objective was to review the major differences of Huntington disease (HD) in Asian population from those in the Caucasian population.

Data Sources:

Data cited in this review were obtained from PubMed database and China National Knowledge Infrastructure (CNKI) from 1994 to 2014. All the papers were written in English or Chinese languages, with the terms of Asia/Asian, HD, genotype, epidemiology, phenotype, and treatment used for the literature search.

Study Selection:

From the PubMed database, we included the articles and reviews which contained the HD patients’ data from Asian countries. From the CNKI, we excluded the papers which were not original research. Due to the language''s restrictions, those data published in other languages were not included.

Results:

In total, 50 papers were cited in this review, authors of which were from the mainland of China, Japan, India, Thailand, Taiwan (China), Korea, and western countries.

Conclusions:

The lower epidemiology in Asians can be partly explained by the less cytosine-adenine-guanine repeats, different haplotypes, and CCG polymorphisms. For the physicians, atypical clinical profiles such as the initial symptom of ataxia, movement abnormalities of Parkinsonism, dystonia, or tics need to be paid more attention to and suggest gene testing if necessary. Moreover, some pathogenesis studies may help progress some new advanced treatments. The clinicians in Asian especially in China should promote the usage of genetic testing and put more effects in rehabilitation, palliative care, and offer comfort of patients and their families. The unified HD rating scale also needs to be popularized in Asia to assist in evaluating the progression of HD.     

Ultra-microstructural changes in iliac artery after bare and magnetic stent implantation in rabbits

Objective To investigate the preventive effect of magnetic stent on coronary restenosis after percutaneous arterial stenting. Methods Twenty rabbits were divided randomly into 2 groups.Bare stent (BS group,n=10) or magnetic stent (MS group,n=10) was implanted in the left iliac artery of the rabbits of the 2 groups,respectively.Aspirin (25rag,qd) was administered orally to the rabbits of both groups from 3 days before stenting until the rabbits were executed.Unfractionated heparin (2500u,qd) was delivered subcuta- neously after stenting for 7 days.Five rabbits of each group were randomly selected to be executed at 7 or 30 days.Structural changes in the injured arteries were studied by optical microscopey,transmissive electronic microscopey and immunohistochemistry.Results At 7 days,more myofibroblasts were found migrating from adventitia to tunica media and intima in BS group than in MS group.Inside the media and intima,large amount of smooth muscle cells of synthetic type were observed.At 30 days after stenting,in magnetic group, most uascular smooth muscle cells (SMCs) under the intima had transformed to contractile type and only little extracellular matrix (ECM) was observed around the SMCs;whereas,in BS group,the SMCs remained to be synthetic type and large amount of ECM was observed around the SMCs,which was composed mainly ofproteoglycans and glycoproteins.Conclusions Magnetic stent can inhibit proliferation and migration of SMCs and reducing the production of ECM,and therefore,may prevent restenosis after coronary stenting.     

Wnt/β-catenin信号通路在阿尔茨海默病神经元变性中的研究进展

目的：阿尔茨海默病(Alzheimer’s disease,AD)是目前老年人最常见的痴呆类型,由于AD发病机制复杂,迄今尚未完全阐明。AD最重要的神经病理学特征是β淀粉样蛋白（amyloid-beta eptide, Aβ）沉积、Tau蛋白过度磷酸化形成神经纤维缠结以及脑内神经炎症反应的激活,并伴随着神经元的损伤及学习记忆功能受损。越来越多的研究指出Wnt/β-连环蛋白（Wnt/β-catenin）信号通路在神经退行性疾病中,尤其是AD中发挥了重要作用。为明确Wnt/β-catenin信号通路与AD发病、Aβ沉积、Tau 蛋白和神经炎症的相关性及其机制,对相关文献进行综述。方法：选择Wnt/β-catenin信号通路与AD、Aβ神经毒性、Tau 蛋白磷酸化及神经炎症相关的国内外相关文献进行综述。结果：研究表明Wnt/β-catenin信号通路参与AD的发生和发展,当该通路激活时可以抑制Aβ的沉积、Tau的过度磷酸化及神经炎症,反之则促进Aβ的产生、聚集以及Tau蛋白的磷酸化,神经炎症的发生。结论：本文就Wnt/β-catenin信号途径在AD发病中的重要性加以概述,为AD机制的研究提供了理论基础,也提示激活Wnt信号传导途径可以作为治疗AD的潜在治疗靶点。dismutase,SOD）、丙二醛（malondialdehyde,MDA）和过氧化氢酶（catalase,CAT）水平采用试剂盒检测。qPCR和Western blot检测血浆Klotho的表达。Klotho表达量与氧化应激的相关性采用Pearson相关性分析。甲基化特异性PCR（MS-PCR）检测Klotho甲基化水平。CpG岛甲基化率采用焦磷酸盐测序法。结果：相对于对照组[（94.56±16.40） mU/L],观察组SOD含量[（79.86±18.24） mU/L]明显降低（t=4.487,P=0.000）,观察组MDA水平[（2.87±2.26） pg/mL]与对照组[（2.52±1.06） pg/mL]无统计学差异（t=1.675,P=0.097）,观察组CAT水平[（18.50±4.62） U/mg]与对照组[（25.26±3.54） U/mg]相比明显降低（t=8.605,P=0.000）。观察组（0.66±0.16）比对照组（1.04±0.10）血浆Klotho基因（t=14.93,P=0.000）降低;观察组（0.70±0.20）比对照组（1.04±0.10）血浆Klotho蛋白表达（t=10.92,P=0.000）降低,且Klotho蛋白表达量与SOD水平（r=0.768,P=0.000）和CAT水平（r=0.708,P=0.000）呈正相关。观察组Klotho启动子CpG岛甲基化水平升高。结论：Klotho甲基化可能是NSCLC患者血浆氧化应激状态的一个临床标志物。     

远端缺血后适应通过抑制RAGE的表达实现对心肌的保护作用

目的：探讨晚期糖基化终末产物受体（receptor for advanced glycation end products,RAGE）在远端缺血后适应改善心肌缺血再灌注中的作用。方法：将34只8~9周龄C57BL/6J小鼠随机分为6组：假手术组（Sham组）、FPS?ZM1对照组（FZM1组）、缺血再灌注组（IR组）、FPS?ZM1干预组（FZM1+IR组）、远端缺血后适应组（RIPostC+IR组）和FZM1+远端缺血后适应干预组（FZM1+RIPostC+IR组）。通过结扎左冠状动脉,建立小鼠心肌缺血再灌注模型。采用小鼠心超仪检测小鼠左室射血分数（left ventricular ejection fraction,LVEF）和左室缩短分数（left ventricular fractional shortening,LVFS）,运用酶联免疫分析（ELISA）法和蛋白印迹法检测炎症因子白介素6（interleukin 6,IL?6）、核因子（nudear factor,NF)?κB P65蛋白、RAGE蛋白。结果：①与IR组相比,RIPostC能显著增加LVEF（P < 0.01）和LVFS（P < 0.01）。②与IR组相比,RIPostC能显著降低RAGE（P < 0.001）的表达。③与IR组相比,RIPostC能显著减少NF?κB P65（P < 0.001）和IL?6（P < 0.001）的表达。结论：RIPostC对心肌缺血再灌注损伤具有保护作用。该作用可能通过抑制RAGE的表达进而降低炎性反应而实现。     

重症手足口病患儿合并肺炎支原体感染实验室检测指标对于临床诊断及治疗的提示性作用

目的 探讨肠道病毒71型合并肺炎支原体感染的重症手足口患儿的检测指标对于临床诊断及治疗是否起到提示作用。方法 收集2014年3月至8月于首都医科大学附属北京地坛医院住院的手足口病患儿100例,非手足口病患儿50例,对其粪便标本采用荧光定量RT-PCR方法检测,得出核酸定性结果。对其血清标本采用被动凝集法检测以得出肺炎支原体抗体效价。分析肺炎支原体感染在手足口病组和非手足口病组中的差异,探讨肠道病毒71型合并肺炎支原体感染与手足口病严重程度的相关性。结果 重症手足口病组患儿的肺炎支原体阳性率(76%)高于轻症手足口病组患儿的肺炎支原体阳性率(48%),差异有统计学意义(χ²=8.319,P=0.004)。结论 肠道病毒71型阳性合并肺炎支原体感染的重症手足口病患儿,应在临床诊治中引起关注。对于降低重症手足口病患儿向危重症的转化具有提示意义。     

大鼠神经元型一氧化氮合酶基因慢病毒载体的构建及功能测定

目的：构建编码大鼠神经元型一氧化氮合酶（neuronal nitric oxide synthase,nNOS）全长基因的慢病毒载体,并检测其表达效率和催化活性功能。方法：采用RT?PCR提取nNOS的cDNA,同时改造真核表达载体pCDH?GFP。鉴定nNOS的cDNA碱基序列正确后,将目的基因克隆入慢病毒载体pCDH?GFP,得重组载体pCDH?GFP/nNOS,采用Lipofectamine 2000将其及慢病毒包装辅助质粒转染293T细胞,超速离心纯化后,感染神经干细胞和海马齿状回神经元进行感染能力鉴定,并检测nNOS蛋白表达和催化功能。结果：成功构建编码nNOS全长cDNA,测序证明重组慢病毒载体pCDH?GFP/nNOS构建成功。包装慢病毒颗粒LV?nNOS?GFP可以感染离体神经干细胞和在体神经元。Western blot检测证明nNOS蛋白成功表达。一氧化氮浓度检测表明表达的nNOS具有催化活性。结论：慢病毒载体LV?nNOS?GFP构建成功,可以表达功能性全长nNOS蛋白。     

膝关节MRI错位征在预测髌骨脱位患者股骨前倾角中的临床应用

目的：建立一个新的影像学标志以预测髌骨脱位患者是否合并过大的股骨前倾角。方法：回顾性分析2016年1月至2018年7月于我院就诊的35例髌骨脱位患者的临床资料。男4例,女31例。平均年龄21.3岁。纳入患者的股骨前倾角为28.3°±8.3°,胫骨扭转角为29.0°±8.9°,股胫参数-1.3±11.1。将膝关节MRI的错位征定义为股骨髁间窝外侧壁与胫骨平台外侧髁间嵴的错位。膝关节MRI错位征阳性层面数为1.49±1.25。结果：纳入的35名患者中有25人出现错位征。错位征阳性与阴性的组别中股骨前倾角存在显著性差异（P<0.05）。股骨前倾角的数值则与错位征在膝关节MRI出现的层面数量呈显著相关（P<0.05）。结论：错位征可见于髌骨脱位患者膝关节MRI中,并且可作为预测患者是否合并股骨前倾角过大的影像学指标之一。其股骨前倾角的数值可通过膝关节MRI中错位征出现的层面数加以预测。     

GDF-5基因重组腺病毒修复人退变髓核细胞的实验研究

【摘要】 目的 探讨腺病毒介导的GDF 5基因对人退变髓核细胞外基质表达的影响,探求一种基因治疗新途径。方法 利用腺病毒载体将GDF 5基因导入退变髓核细胞,设置为空白对照组（Control组）、阴性对照组（GFP组）、实验组（GDF 5组）3组,并分为3、7、14、21天四个观察时间点,分别检测3组髓核细胞sGAG、Hyp的含量,免疫染色、Safranine O染色观察细胞外基质合成情况,Real time PCR检测Aggrecan、Collagen II mRNA表达水平。结果 GDF 5组髓核细胞sGAG/DNA、Hyp/DNA均在14、21天较Control组和GFP组明显增加,差异有统计学意义（P＜0.05）,而Control组与GFP组之间任意观察时间点比较差异均无统计学意义（P＞0.05）;免疫染色、Safranine O染色结果显示GDF 5组髓核细胞的蛋白多糖、Ⅱ型胶原、蛋白聚糖在14、21天均呈强阳性染色,Control组和GFP组呈弱阳性染色。Real time PCR结果显示GDF 5组Aggrecan、Collagen II mRNA的表达在14、21天较Control组和GFP组明显增加,差异有统计学意义（P＜0.05）。结论 腺病毒介导的GDF 5能明显促进人退变髓核细胞Aggrecan、Collagen II mRNA的表达,并增加蛋白多糖、Ⅱ型胶原的合成,对细胞外基质具有明确的修复作用,是一种有效的基因治疗新方法。