超声微泡介导微RNA治疗肝细胞癌的研究进展

基因治疗正逐渐成为肝细胞癌（HCC）、尤其晚期HCC的治疗方案。足够剂量的特定微RNA（miRNA）可抑制HCC细胞生长,超声微泡介导基因和药物递送治疗策略可促使miRNA于肿瘤区域靶向释放,从而达到治疗效果。本文对超声微泡介导miRNA治疗HCC研究进展进行综述。     

The emerging role of microRNAs in regulating immune and inflammatory responses in the lung

Chronic inflammatory diseases of the lung are leading causes of morbidity and mortality worldwide. Many of these disorders can be attributed to abnormal immune responses to environmental stimuli and infections. As such, understanding the innate host defense pathways and their regulatory systems will be critical to developing new approaches to treatment. In this regard, there is increasing interest in the role of microRNAs (miRNAs) in the regulation of pulmonary innate host defense responses and the inflammatory sequelae in respiratory disease. In this review, we discuss recent findings that indicate an important role for miRNAs in the regulation in mouse models of various respiratory diseases and in host defense against bacterial and viral infection. We also discuss the potential utility and limitations of targeting these molecules as anti-inflammatory strategies and also as a means to improve pathogen clearance from the lung.     

MicroRNAs in laser-induced choroidal neovascularization in mice and rats: their expression and potential therapeutic targets

Choroidal neovascularization characterizes wet age-related macular degeneration. Choroidal neovascularization formation involves a primarily angiogenic process that is combined with both inflammation and proteolysis. A primary cause of choroidal neovascularization pathogenesis is alterations in pro-and anti-angiogenic factors derived from the retinal pigment epithelium, with vascular endothelium growth factor being mainly responsible for both clinical and experimental choroidal neovascularization. MicroRNAs(miRNAs) which are short, non-coding, endogenous RNA molecules have a major role in regulating various pathological processes, including inflammation and angiogenesis. A review of recent studies with the mouse laser-induced choroidal neovascularization model has shown alterations in miRNA expression in choroidal neovascularization tissues and could be potential therapeutic targets for wet age-related macular degeneration. Upregulation of miR-505(days 1 and 3 post-laser), miR-155(day 14) occurred in retina; miR-342-5 p(days 3 and 7), miR-126-3 p(day 14) in choroid; miR-23 a, miR-24, miR-27 a(day 7) in retina/choroid; miR-505(days 1 and 3) in retinal pigment epithelium/choroid; downregulation of miR-155(days 1 and 3), miR-29 a, miR-29 b, miR-29 c(day 5), miR-93(day 14), miR-126(day 14) occurred in retinal pigment epithelium/choroid. Therapies using miRNA mimics or inhibitors were found to decrease choroidal neovascularization lesions. Choroidal neovascularization development was reduced by overexpression of miR-155, miR-188-5 p, miR-(5,B,7), miR-126-3 p, miR-342-5 p, miR-93, miR-126, miR-195 a-3 p, miR-24, miR-21, miR-31, miR-150, and miR-184, or suppression of miR-505, miR-126-3 p, miR-155, and miR-23/27. Further studies are warranted to determine miRNA expression in mouse laser-induced choroidal neovascularization models in order to validate and extend the reported findings. Important experimental variables need to be standardized; these include the strain and age of animals, gender, number and position of laser burns to the eye, laser parameters to induce choroidal neovascularization lesions including wavelength, power, spot size, and duration.     

JCL roundtable: Lipids and inflammation in atherosclerosis

Clinical effort in lipidology focuses largely on mitigating effects of atherosclerosis, a pathologic process localized to the intimal layer of larger arteries. This JCL Roundtable brings together 3 leading researchers to discuss the current understanding of pathogenesis in atherosclerosis. We begin by recognizing that low density lipoprotein concentrations in arterial intima far exceed concentrations in other connective tissues, consistent with the response-to-retention hypothesis of atherogenesis. High density lipoproteins facilitate reverse cholesterol transport and also have antioxidant and anti-inflammatory roles. New evidence points to remnants of triglyceride-rich lipoproteins as promoters of atherogenesis, highlighted by deleterious effects of apolipoprotein C-III. The multifaceted role of inflammation is becoming clearer through discoveries related to leukocyte recruitment, efferocytosis, resolution of inflammation, and crystal formation. MicroRNAs represent a new, complex mode of gene regulation bearing on lipoprotein and inflammation biology. Progress in understanding atherosclerosis portends a future in which residual risk related to obesity, diabetes, and other factors will yield to new targeted therapies.     

CRISPR系统与哺乳动物免疫系统的比较及其功能的研究进展

原核生物防御噬菌体入侵的机制与哺乳类免疫防御机制有很多相似之处,尤其是特异性免疫机制,原核生物的特异性免疫系统即成簇规律间隔短回文重复序列（ clustered regularly interspaced short palindromic repeats, CRISPR）与哺乳类特异性免疫系统在很多环节体现了相似的规律。通过比较,能够加深对CRISPR的免疫功能的认识。近年发现,除了发挥特异性免疫功能,CRISPR系统还参与了许多细菌生长代谢过程的调控机制,调控基因表达的能力更是受到广泛的关注。 CRISPR系统功能的复杂程度远远超出免疫防御的范畴,值得深入研究。     

血浆microRNA实时荧光定量PCR检测方法的建立

目的建立特异、稳定且可靠的血浆microRNA(miRNAs)实时荧光定量PCR技术。方法收集10例健康人血浆标本,采用mirVanaTM PARIS kit试剂盒法提取血浆miRNAs,以miRNA特异的茎环引物引导逆转录,通过SYBR Green实时荧光定量聚合酶链反应(PCR)对外参照cel-miR-39、cel-miR-238及血浆miR-342-3p进行检测。结果健康人血浆miR-342-3p及外参cel-miR-39、cel-miR-238可以实现特异性扩增及定量。溶解曲线显示10例健康人血浆标本中cel-miR-39、cel-miR-238和miR-342-3p扩增产物分别在81.44、81.62、82.71℃时出现单一峰,无引物二聚体峰和其他非特异峰出现。血浆miR-342-3p批内批间重复性实验的标准差在0.13~0.20、变异系数(CV)在0.42%~0.66%,显示该方法重复性较好。以cel-miR-39、cel-miR-238作为外参照,同一血浆标本miR-342-3p的5次检测结果ΔCt的标准差为0.22、CV为1.68%,说明cel-miR0-39、cel-miR-238可作为血浆miRNAs实时荧光定量PCR检测的稳定外参。结论实时荧光定量PCR技术可作为研究血浆miRNAs较好的技术平台。     

放射后鼻咽癌细胞株C666-1存活亚克隆高表达miR-181a

目的：：初步分析miR-181a与鼻咽癌细胞系C666-1放射敏感的相关性。方法：首先使用大剂量8 Gy的X射线照射C666-1,挑选三个大的存活亚克隆并命名为 C666-1-R1,C666-1-R2和 C666-1-R3,然后应用MTT等技术分析亚克隆及对照母本C666-1细胞系放射敏感性的差异,最后采用实时荧光定量PCR 法,分析三个存活亚克隆及对照C666-1的miR-181 a表达水平。结果：通过MTT比色法检测细胞增殖：6 Gy照射后,三个亚克隆C666-1-R1,C666-1-R2和C666-1-R3的细胞存活率均高于对照细胞C666-1,尤其C666-1-R2在48、72和96 h的存活率增高均有显著性差异(P<0.05),提示C666-1-R2具有放射抗拒性。另一方面,各亚克隆及对照细胞的 miR-181a 的2-△△Ct值如下：C666-1-R1=0.693,C666-1-R2=0.486,C666-1-R3=0.762,C666-1=1。提示放射后存活的三个亚克隆均高表达 miR-181a,尤其放射抗拒亚克隆C666-1-R2的 miR-181a表达更高。结论：miR-181a与鼻咽癌存活亚克隆的放射抗拒性密切相关。miR-181a可能是评估鼻咽癌放射敏感性的一个新分子,miR-181a 与鼻咽癌放射抗拒的相关性值得深入研究。     

心房颤动患者冠状窦血小分子RNA表达差异与价值

目的探讨非瓣膜性心房颤动(房颤)患者冠状窦血小分子RNA(microRNA,miRNA)表达差异及其调控房颤的机制,早期预警诊断和干预的价值。方法选择15例房颤患者作为房颤组,其中阵发性房颤、持续性房颤和永久性房颤各5例,选择健康体检者5例作为正常对照组。导管射频消融术前和术中分别取外周血和冠状窦血,使用miRNA芯片进行全基因组miRNA表达谱微阵列分析,Volcano Plot法获得差异表达miRNA,通过mirbase、miranda、targetscan数据库进行靶基因分析。结果房颤组自身冠状窦血与外周血比较,有14个miRNA表达差异显著,其中6个表达上调:miR-1266、miR-4279、miR-4787-5p、miR-4666a-3p、kshv-miR-K12-6-3p、miR-3150a-5p;8个表达下调:miR-892a、miR-3149、miR-3171、miR-3664-5p、miR-3591-3p、miR-4423-5p、miR-4473、miR-574-3p。miR-1266在阵发性房颤、持续性房颤和永久性房颤患者均明显升高,而miR-3171则显著降低。房颤组冠状窦血和外周血与正常对照组比较,miRNA表达有明显差异(P<0.05)。结论房颤患者冠状窦血更能反映心脏miRNA的代谢与调控状况;外周血miR-3171、miR-892a、miR-3149在房颤发生早期出现,且持续差异表达,有可能成为房颤诊断的标记物;miR-1266、miR-4279、miR-4666a-3p有可能成为未来房颤治疗的干预靶点。