早期应用生长反应因子1小干扰RNA可抑制烟草烟雾刺激大鼠主动脉平滑肌细胞白细胞介素1β mRNA的表达

背景：研究表明,在动脉粥样硬化的血管壁细胞中白细胞介素1β表达水平升高。目的：观察早期生长反应因子1的小干扰RNA对烟草烟雾提取物刺激大鼠主动脉平滑肌细胞中白细胞介素1β mRNA表达的影响。方法：体外培养大鼠主动脉平滑肌细胞,用不同体积分数（0%,5%,10%,20%和40%）烟草烟雾提取物刺激细胞,筛选最佳体积分数烟草烟雾提取物干预细胞0,8,16和24h,以确定最佳干预时间,最后用生长反应因子1小干扰RNA以沉默细胞中生长反应因子1的表达。结果与结论：RT-PCR结果显示,烟草烟雾提取物可诱导大鼠主动脉平滑肌细胞白细胞介素1β mRNA的表达,且有时间和浓度依赖性,烟草烟雾提取物最佳干预时间为8h,最佳干预体积分数为10%。将生长反应因子1小干扰RNA转染10%烟草烟雾提取物刺激的细胞后8h,白细胞介素1β mRNA表达明显减少（P〈0.05）。证实早期应用生长反应因子1小干扰RNA可以抑制烟草烟雾提取物诱导的大鼠主动脉平滑肌细胞中白细胞介素1β mRNA的表达。     

早期应用生长反应因子1小干扰RNA可抑制烟草烟雾刺激大鼠主动脉平滑肌细胞白细胞介素1β mRNA的表达

背景:研究表明,在动脉粥样硬化的血管壁细胞中白细胞介素1β表达水平升高。目的:观察早期生长反应因子1的小干扰RNA对烟草烟雾提取物刺激大鼠主动脉平滑肌细胞中白细胞介素1β mRNA表达的影响。方法:体外培养大鼠主动脉平滑肌细胞,用不同体积分数(0%,5%,10%,20%和40%)烟草烟雾提取物刺激细胞,筛选最佳体积分数烟草烟雾提取物干预细胞0,8,16和24h,以确定最佳干预时间,最后用生长反应因子1小干扰RNA以沉默细胞中生长反应因子1的表达。结果与结论:RT-PCR结果显示,烟草烟雾提取物可诱导大鼠主动脉平滑肌细胞白细胞介素1β mRNA的表达,且有时间和浓度依赖性,烟草烟雾提取物最佳干预时间为8h,最佳干预体积分数为10%。将生长反应因子1小干扰RNA转染10%烟草烟雾提取物刺激的细胞后8h,白细胞介素1β mRNA表达明显减少(P<0.05)。证实早期应用生长反应因子1小干扰RNA可以抑制烟草烟雾提取物诱导的大鼠主动脉平滑肌细胞中白细胞介素1β mRNA的表达。     

香烟提取物影响大鼠主动脉平滑肌细胞结缔组织生长因子的表达

背景：血管平滑肌细胞增殖是心血管系统组织变化的基础。目的：观察不同浓度香烟提取物对大鼠主动脉平滑肌细胞结缔组织生长因子表达的影响。方法：取第3代大鼠主动脉平滑肌细胞,分别用体积分数2.5%,5%,10%,20%的香烟提取物进行干预。结果与结论：实时定量RT-PCR结果显示,体积分数2.5%,5%,10%,20%的香烟提取物作用12h均可促进大鼠主动脉平滑肌细胞结缔组织生长因子mRNA的表达,以体积分数5%香烟提取物的作用最明显,且其刺激大鼠主动脉平滑肌细胞结缔组织生长因子mRNA的表达在24h内呈时间依赖性。提示香烟提取物可上调大鼠主动脉平滑肌细胞结缔组织生长因子mRNA的表达,导致大鼠主动脉平滑肌细胞的增殖。     

香烟提取物影响大鼠主动脉平滑肌细胞结缔组织生长因子的表达

背景:血管平滑肌细胞增殖是心血管系统组织变化的基础。目的:观察不同浓度香烟提取物对大鼠主动脉平滑肌细胞结缔组织生长因子表达的影响。方法:取第3代大鼠主动脉平滑肌细胞,分别用体积分数2.5%,5%,10%,20%的香烟提取物进行干预。结果与结论:实时定量RT-PCR结果显示,体积分数2.5%,5%,10%,20%的香烟提取物作用12h均可促进大鼠主动脉平滑肌细胞结缔组织生长因子mRNA的表达,以体积分数5%香烟提取物的作用最明显,且其刺激大鼠主动脉平滑肌细胞结缔组织生长因子mRNA的表达在24h内呈时间依赖性。提示香烟提取物可上调大鼠主动脉平滑肌细胞结缔组织生长因子mRNA的表达,导致大鼠主动脉平滑肌细胞的增殖。     

血管平滑肌细胞凋亡与阿托伐他汀的干预效应

目的：观察阿托伐他汀对血管平滑肌细胞凋亡的影响,并探讨凋亡抑制因子Survivin蛋白表达的意义.方法：实验于2004-04/2005-03在医学遗传学国家重点实验室中南大学湘雅二医院中心实验室完成.①取培养五六代的5只1月龄健康雄性大鼠主动脉中层平滑肌细胞,进行干预前于六孔培养板中放置无菌盖玻片,细胞长满盖玻片的90%时开始干预,分为4组进行干预：正常对照组：培养的平滑肌细胞未用阿托伐他汀干预;阿托伐他汀0.1μmol/L组：用浓度为0.1μmol/L的阿托伐他汀干预培养的平滑肌细胞;阿托伐他汀1.0μmol/L组：用浓度为1.0 μmol/L的阿托伐他汀干预培养的平滑肌细胞;阿托伐他汀10.0 μmol/L组：用浓度为10.0μmol/L的阿托伐他汀干预培养的平滑肌细胞.②分别于干预后6,12,24,48,72 h采用Hoechst 33258荧光染色观察血管平滑肌细胞凋亡情况：用免疫组化法检测血管平滑肌细胞凋亡抑制因子Survivin蛋白表达情况.③组间差异比较采用t检验.结果：①荧光染色检测血管平滑肌细胞凋亡结果显示,阿托伐他汀干预培养的大鼠血管平平滑肌细胞后,随干预时间延长和用药浓度增加,平滑肌细胞凋亡增加.阿托伐他汀干预72 h后,血管平滑肌细胞凋亡率：阿托伐他汀0.1,1.0,10.0 μmol/L组明显高于正常对照组（11.1%,27.2%,50.5%,3.8%,P＜0.01）,各组间两两比较差异明显（P＜0.01）.②培养的血管平滑肌细胞Survivin因子表达情况：阿托伐他汀干预24 h,血管平滑肌细胞凋亡抑制因子Survivin蛋白表达下降,干预48 h～72 h,血管平滑肌细胞Survivin因子未见表达.结论：①阿托伐他汀可诱导血管平滑肌细胞凋亡,其凋亡率呈时间和剂量依赖性.②阿托伐他汀致血管平滑肌细胞凋亡的作用可能与其下调凋亡抑制因子Survivin蛋白的表达有关.     

RNA干扰选择性下调血管平滑肌细胞结缔组织生长因子的表达

目的:从血管平滑肌细胞结缔组织生长因子的基础分泌及转化生长因子β1对其分泌的诱导作用入手,进一步观察并验证RNA干扰技术靶向抑制结缔组织生长因子对下游纤维化因子的影响。方法:实验于2006-03/2007-05在武汉协和医院心血管病研究所实验室完成。①实验材料:健康雄性成年SD大鼠,体质量150～180g。②实验方法及分组:分离培养成年大鼠血管平滑肌细胞,以5μg/L终浓度转化生长因子β1对培养的血管平滑肌细胞进行诱导,再通过脂质体法将靶向大鼠结缔组织生长因子的shRNA干扰质粒转染入血管平滑肌细胞,200mg/L终浓度的G418筛选出稳定转染细胞株后培养增殖进入后续实验。将实验分为5组,分别为对照组,不给与任何干预措施;转化生长因子β1刺激组,在培养的血管平滑肌细胞培养液中加入转化生长因子β1(5μg/L终浓度)刺激诱导24h;HK阴性质粒对照组,血管平滑肌细胞常规培养3～5代后转染HK阴性质粒继续培养48h;RNA干扰组,先以转化生长因子β1(5μg/L终浓度)诱导刺激血管平滑肌细胞24h后再转染干扰质粒继续培养48h;单纯转染试剂组,细胞常规培养3～5代后仅加入lipo2000转染试剂。③实验评估:通过逆转录-聚合酶链反应和(或)蛋白免疫印迹技术进一步检测各组心肌细胞结缔组织生长因子、纤维连接蛋白mRNA及蛋白的表达。结果:①组织块贴附法培养的原代血管平滑肌细胞,可见大量细胞紧密排列成长梭形。②倒置荧光显微镜下观察,脂质体lipo2000转染血管平滑肌细胞瞬时转染效率仅有大约30%左右;而通过加入G418进行稳定转染后,几乎所有细胞均可被激发出亮绿色荧光,转染效率可达95%以上。③对照组结缔组织生长因子、纤维连接蛋白均有低水平的基础分泌,在予以转化生长因子β1诱导后表达显著升高(P<0.01);而在靶向特异性RNA干扰后,与转化生长因子β1刺激组比较,结缔组织生长因子及纤维连接蛋白的表达均被显著抑制(P<0.01);并且纤维化因子纤维连接蛋白及胶原的表达与促纤维化因子结缔组织生长因子的表达有显著相关性。结论:血管平滑肌细胞可自主分泌结缔组织生长因子及细胞外基质成分;而通过RNA干扰靶向抑制结缔组织生长因子后可阻抑血管平滑肌细胞分泌下游纤维化因子。提示血管平滑肌细胞可主动参与心血管重塑及纤维化,而RNA干扰靶向沉默结缔组织生长因子基因的表达可以有效干预纤维化过程。     

内皮型一氧化氮合酶基因转染对血管损伤后新生内膜增生的影响

背景：一氧化氮能够抑制血管平滑肌细胞的迁移和增殖,而一氧化氮合酶是其合成的关键酶,有关一氧化氮合酶基因体内转染对平滑肌细胞及动脉粥样硬化血管损伤后内膜增生影响少有报道。目的：观察内皮型一氧化氮合酶（endothelial nitric oxide synthase,eNOS）基因体内局部转染对动脉粥样硬化大鼠血管损伤后新生内膜增生的抑制作用。方法：建立动脉粥样硬化Wistar大鼠颈动脉球囊损伤模型,建模后随机分成空白对照组、AdCMV-lacz对照组和AdCMV-eNOS组,分别将PBS,AdCMV-lacz和AdCMV-eNOS体内转染至以上3组大鼠的损伤血管壁。转染2周后培养并鉴定损伤局部平滑肌细胞,并用RT-PCR法检测各组损伤及转染后血管平滑肌细胞eNOS mRNA的表达,同时观察转染后不同时期新生内膜增生的影响。结果与结论：AdCMV-eNOS组的颈总动脉血管平滑肌细胞可表达eNOS mRNA。3组大鼠转染后1和3个月,AdCMV-eNOS组内膜/中膜面积比值低于空白对照组和AdCMV-lacz对照组（P〈0.01）。结果显示,eNOS基因体内转染损伤后血管可以抑制血管新生内膜增生,减少再狭窄发生率。     

生长反应因子与大鼠颈动脉损伤后内膜增生

背景：经皮冠状动脉介入治疗后再狭窄仍是影响介入治疗远期疗效的严重的临床问题。目的：在大鼠颈动脉球囊损伤模型中,探讨早期生长反应因子诱骗寡脱氧核苷酸对损伤后的血管内膜的影响,并初步探讨其分子机制。方法：利用2FFogarty导管损伤Wistar大鼠颈动脉,构建大鼠球囊损伤模型,在转染试剂Fugene6介导下经血管腔内转染生长反应因子诱骗寡脱氧核苷酸,苏木精一伊红染色法观察血管内膜增生情况,免疫组化法检测CyclinD1,观察其表达情况。结果与结论：早期生长反应因子诱骗寡脱氧核苷酸可以显著抑制大鼠颈动脉球囊损伤后血管内膜增生,同时也可以下调大鼠颈动脉球囊损伤后表达显著增加的CyclinD1。说明生长反应因子诱骗寡脱氧核苷酸可能通过抑制CyclinD1的表达,使细胞周期停滞,从而抑制损伤后的大鼠颈动脉内膜的增生。     

基质细胞衍生因子1α及受体CXCR4对颈总动脉球囊损伤血管平滑肌细胞和骨髓基质细胞的影响

背景：在细胞因子的作用下,骨髓基质细胞可以向内皮祖细胞分化,并迁移至损伤部位增殖,参与血管新生内膜的修复。目的：观察基质细胞衍生因子1α及其受体CXCR4在大鼠颈总动脉球囊损伤后对血管平滑肌细胞和骨髓基质细胞的影响。方法：体外培养大鼠颈总动脉球囊损伤后各时间点（即刻、1,4,7,14及1个月）的血管平滑肌细胞和骨髓基质细胞,检测损伤后平滑肌细胞中基质细胞衍生因子1α及骨髓基质细胞中CXCR4的表达情况,并应用基质细胞衍生因子1αsiRNA转染损伤后血管平滑肌细胞,同时应用transwell小室观察基质细胞衍生因子1α和损伤后血管平滑肌细胞对骨髓基质细胞的趋化作用。结果与结论：大鼠颈总动脉球囊损伤后,平滑肌细胞中基质细胞衍生因子1αmRNA表达开始增加,至损伤第7天后逐渐减弱;血管损伤后4d起,骨髓基质细胞中CXCR4mRNA开始出现,并持续表达;transwell小室显示损伤后的骨髓基质细胞对基质细胞衍生因子1α的趋化性增强;CXCR4受体拮抗剂AMD3100可以明显减弱基质细胞衍生因子1α对骨髓基质细胞的趋化作用;损伤后的血管平滑肌细胞对骨髓基质细胞的趋化作用强于基质细胞衍生因子1α（P〈0.05）,应用AMD3100干预或细胞内转染基质细胞衍生因子1αsiRNA后,此趋化作用亦减弱（P〈0.05）。结果提示,基质细胞衍生因子1α/CXCR4轴在骨髓基质细胞向损伤血管迁移中起重要的作用。     

恒磁场对大鼠主动脉平滑肌细胞骨桥蛋白基因表达的影响

背景:适当强度的恒磁场可抑制血管平滑肌细胞增殖,可能用于抑制经皮冠状动脉介入治疗术后再狭窄。目的:观察不同磁感应强度恒磁场对培养的大鼠主动脉血管平滑肌细胞骨桥蛋白基因表达的影响,以探讨磁场是否能用于经皮冠状动脉介入治疗术后再狭窄的防治。设计:随机分组,对照观察。单位:解放军第四军医大学西京医院心内科。材料:实验于2006-02/12在解放军第四军医大学西京医院心内科实验室(全军心血管病研究所)完成。纯种雄性SD大鼠,体质量200～250g。方法:用含体积分数为0.1小牛血清的DMEM培养液体外培养大鼠主动脉血管平滑肌细胞,实验随机分为对照组,1Gs恒磁场组、5Gs恒磁场组、10Gs恒磁场组、50Gs恒磁场组,其中对照组不予磁场干预,其他各组分别给予磁场干预继续培养48h。应用反转录-聚合酶链反应及蛋白免疫印迹技术结合吸光度扫描分析,观察恒磁场对血管平滑肌细胞骨桥蛋白及骨桥蛋白mRNA表达的影响。主要观察指标:各组血管平滑肌细胞骨桥蛋白及骨桥蛋白mRNA的表达。结果:对照组细胞表达一定量的骨桥蛋白及骨桥蛋白基因,各恒磁场组均较对照组降低,差异显著(P<0.05)。不同磁场强度组间分析显示,恒磁场的刺激作用具有磁场强度依赖性,随磁场强度加大,抑制骨桥蛋白及骨桥蛋白mRNA表达作用增强。结论:适当强度的恒磁场能在基因水平上抑制血管平滑肌细胞骨桥蛋白的表达,磁场对经皮冠状动脉介入治疗术后的再狭窄可能具有防治作用。     

儿童遗传性心律失常的诊治进展

回顾在遗传性心律失常领域最新发表的相关研究,主要关注与儿童心源性猝死关系密切的离子通道病,包括长QT综合征（LQTS）、短QT综合征（SQTS）、Brugada综合征（BrS）和儿茶酚胺敏感性多形性室性心动过速（CPVT）,总结它们在发病机制及诊治方面的进展。     

The acoustic characteristics of the skull

Many investigators have stated that the difficulties of imaging with acoustical energy through the skull result from the marked attenuation of the energy by the skull. In the literature measurements of total attenuation have been confused with those for absorption.Measurements made by us show that absorption by compact bone varies between 2–3 dB cm^?1 MHz^?1 and, in the low megaHertz region appears to be directly proportional to frequency.It has also between shown that the convoluted inner surface of the ivory bone of the inner table of the skull may degrade the collimation and directionality of the beam by refraction.Cancellous bone, such as is present in the dipole of the skull, greatly attenuates the energy. It is postulated that this largely results from scattering. It is also postulated that the energy propagates through cancellous bone as two components, one in the soft tissues and the other partly in the bony spicules. Observations suggest that attenuation due to scattering much more markedly affects the latter of these components and scatters more greatly the higher frequencies in a pulse of broad bandwidth.The energy in each component has varying propagation paths so that the later cycles in the pulse of each component are subject to increasing interference as a result of the variations in propagation times. The two components moreover may have different propagation times so that interference may occur between the pulses of each component as well.All of these phenomena degrade the collimation, coherence, directionality, beam width, pulse length, frequency and other properties of the ultrasonic energy upon which imaging through the skull depends.The interference effects described above are least for the first cycle in the pulse which usually is not the cycle of highest amplitude. Since, in the free field, most of the energy is concentrated around the beam axis, most of the energy in the field which is deflected from its normal propagation path is deflected away from the beam axis. Thus the directionality of the beam is least degraded in the beam axis. The effects of the skull in degrading the properties of the ultrasonic pulse would therefore be lessened if the amplitude of the first cycle of the pulse and the directionality of its energy could be used for imaging.     

Organ Preservation: Current Concepts and New Strategies for the Next Decade

SUMMARY: Organ transplantation has developed over the past 50 years to reach the sophisticated and integrated clinical service of today through several advances in science. One of the most important of these has been the ability to apply organ preservation protocols to deliver donor organs of high quality, via a network of organ exchange to match the most suitable recipient patient to the best available organ, capable of rapid resumption of life-sustaining function in the recipient patient. This has only been possible by amassing a good understanding of the potential effects of hypoxic injury on donated organs, and how to prevent these by applying organ preservation. This review sets out the history of organ preservation, how applications of hypothermia have become central to the process, and what the current status is for the range of solid organs commonly transplanted. The science of organ preservation is constantly being updated with new knowledge and ideas, and the review also discusses what innovations are coming close to clinical reality to meet the growing demands for high quality organs in transplantation over the next few years.     

2017年呼吸疾病主要进展

2017年,国内外学者在呼吸系统疾病的临床和基础领域均进行了深入研究,不仅对相关指南进行了更新,并且针对一些临床热点、难点问题达成专家共识,现就2017年呼吸疾病相关进展作一简单介绍。     

家族性噬血细胞性淋巴组织细胞增生症1例并文献复习

目的加强对家族性噬血细胞性淋巴组织细胞增生症（familially hemophagocytic lymphohistiocytosis,FHL）的认识。方法报道确诊为FHL的新病例1例,结合国内外报道的FHL的病例,对该病的临床特点进行汇总分析。结果FHL2常与PRF1基因突变相关,约20%~40%的患者存在穿孔素基因突变。结论对于有阳性家族史,基因诊断明确,应尽早行化疗或者造血干细胞移植。若无家族史,未发现与继发性HLH相关的原发病因,可考虑行基因筛查以明确是否存在FHL的可能。     

COVID‐19 and COVID‐19 vaccine‐related dermatological reactions: An interesting case series with a narrative review of the potential critical and non‐critical mucocutaneous adverse effects related to virus,therapy, and the vaccination

This narrative review article was conducted to lay out a summarized but exhaustive review of current literature over mucocutaneous manifestations in 4 dimensions of SARS‐CoV‐2 pandemic: virus itself, treatment‐related, vaccine‐induced, and alteration of chronic dermatologic diseases following infection. Virus and vaccine‐related were mainly self‐limited and non‐severe. Treatment‐related reactions could be life‐threatening.     

Burkitt's lymphoma of maxillary gingiva: A case report

Burkitt's lymphoma(BL) is an aggressive form of nonHodgkin's B-cell lymphoma with three variants namely endemic, sporadic, and immunodeficiency-associated types. It is endemic in Africa and sporadic in other parts of the world. While the endemic form is widely reported to occur in early childhood and commonly involves the jaw bones, the sporadic form typically presents as an abdominal mass. This presentation reports a rare case of sporadic form of BL clinically manifesting as a generalized gingival enlargement in an immunocompetent adult male which demonstrated an aggressive behavior. The patient reported with a prominent anterior gingival swelling of 6 mo duration which slowly enlarged in size and associated with multiple lymph node involvement. Microscopic examination of the lesion using H, E and immunohistochemical diagnosis confirmed the diagnosis as BL. The patient succumbed to the disease before any therapy could be instituted. Since a wide array of causes can be attributed to gingival enlargements, it is necessary to consider malignancies as one of the important differential diagnosis so as to facilitate the need for appropriate diagnosis and prompt treatment.     

Real‐life effectiveness of biological therapies on symptoms in severe asthma with comorbid CRSwNP

BackgroundWe aimed to evaluate the effectiveness of different antibody therapies on nasal polyp symptoms in patients treated for severe asthma.MethodsWe performed a retrospective analysis of patients with severe asthma and comorbid CRSwNP who were treated with anti‐IgE, anti‐IL‐5/R or anti‐IL‐4R. CRSwNP symptom burden was evaluated before and after 6 months of therapy.ResultsFifty patients were included hereof treated with anti‐IgE: 9, anti‐IL‐5/R: 26 and anti‐IL‐4R: 15 patients. At baseline median SNOT‐20 was similar among groups (anti‐IgE: 55, anti‐IL‐5/R: 52 and anti‐IL‐4R: 56, p = 0.76), median visual analogue scale (VAS) for nasal symptoms was 4, 7 and 8 (p = 0.14) and VAS for total symptoms was higher in the anti‐IL‐4R group (4, 5 and 8, p = 0.002). After 6 months SNOT‐20 improved significantly in all patient groups with median improvement of anti‐IgE: −8 (p < 0.01), anti‐IL‐5/R: −13 (p < 0.001) and anti‐IL‐4R: −18 (p < 0.001), with larger improvement in the anti‐IL‐4R group than in anti‐IgE (p < 0.001) and anti‐IL‐5/R (p < 0.001) groups. VAS nasal symptoms improved by median anti‐IgE: 0 (n.s.), anti‐IL‐5/R: −1 (p < 0.01) and anti‐IL‐4R: −3 (p < 0.001), VAS total symptoms by anti‐IgE: −1 (n.s.), anti‐IL‐5/R: −2 (p < 0.001) and anti‐IL‐4R: −2 (p < 0.001).ConclusionsTreatment by all antibodies showed effectiveness in reducing symptoms of CRSwNP in patients with severe asthma, with the largest reduction observed in anti‐IL‐4R‐treated patients.     

Recalcitrant giant orf recurrence after amputation: A case report and review of the literature

Orf is caused by a parapoxvirus. We present a recurrent, giant digital orf case in a female patient with a history of hairy cell leukemia. In spite of shave excision, the lesion progressed and recurred after digital amputation. Treatment with topical imiquimod cream and systemic subcutaneous interferon alfa‐2a was successful.