诺如病毒反向遗传系统的研究进展与应用

诺如病毒是导致人类急性胃肠炎的主要食源性致病原,由于变异快且缺乏稳健的体外细胞培养体系及小动物感染模型,限制了我们对该病原体的深入研究.近年来,可培养鼠源诺如病毒的发现、人源诺如病毒肠道细胞培养体系的开发,以及诺如病毒反向遗传操作体系的构建为系统研究该病原提供了有力工具,加深了我们对其复制机制、致病机理、病毒-宿主相互...     

Long noncoding RNA lncMREF promotes myogenic differentiation and muscle regeneration by interacting with the Smarca5/p300 complex

Long noncoding RNAs (lncRNAs) play important roles in the spatial and temporal regulation of muscle development and regeneration. Nevertheless, the determination of their biological functions and mechanisms underlying muscle regeneration remains challenging. Here, we identified a lncRNA named lncMREF (lncRNA muscle regeneration enhancement factor) as a conserved positive regulator of muscle regeneration among mice, pigs and humans. Functional studies demonstrated that lncMREF, which is mainly expressed in differentiated muscle satellite cells, promotes myogenic differentiation and muscle regeneration. Mechanistically, lncMREF interacts with Smarca5 to promote chromatin accessibility when muscle satellite cells are activated and start to differentiate, thereby facilitating genomic binding of p300/CBP/H3K27ac to upregulate the expression of myogenic regulators, such as MyoD and cell differentiation. Our results unravel a novel temporal-specific epigenetic regulation during muscle regeneration and reveal that lncMREF/Smarca5-mediated epigenetic programming is responsible for muscle cell differentiation, which provides new insights into the regulatory mechanism of muscle regeneration.     

The novel type II toxin–antitoxin PacTA modulates Pseudomonas aeruginosa iron homeostasis by obstructing the DNA-binding activity of Fur

Type II toxin–antitoxin (TA) systems are widely distributed in bacterial and archaeal genomes and are involved in diverse critical cellular functions such as defense against phages, biofilm formation, persistence, and virulence. GCN5-related N-acetyltransferase (GNAT) toxin, with an acetyltransferase activity-dependent mechanism of translation inhibition, represents a relatively new and expanding family of type II TA toxins. We here describe a group of GNAT-Xre TA modules widely distributed among Pseudomonas species. We investigated PacTA (one of its members encoded by PA3270/PA3269) from Pseudomonas aeruginosa and demonstrated that the PacT toxin positively regulates iron acquisition in P. aeruginosa. Notably, other than arresting translation through acetylating aminoacyl-tRNAs, PacT can directly bind to Fur, a key ferric uptake regulator, to attenuate its DNA-binding affinity and thus permit the expression of downstream iron-acquisition-related genes. We further showed that the expression of the pacTA locus is upregulated in response to iron starvation and the absence of PacT causes biofilm formation defect, thereby attenuating pathogenesis. Overall, these findings reveal a novel regulatory mechanism of GNAT toxin that controls iron-uptake-related genes and contributes to bacterial virulence.     

Endocytosis of Peptidase Inhibitor SerpinE2 promotes Myocardial Fibrosis through activating ERK1/2 and β-catenin Signaling Pathways

Cardiac fibrosis is one of the common pathological processes in many cardiovascular diseases characterized by excessive extracellular matrix deposition. SerpinE2 is a kind of protein that inhibits peptidase in extracellular matrix and up-regulated tremendously in mouse model of cardiac fibrosis induced by pressure-overloaded via transverse aortic constriction (TAC) surgery. However, its effect on cardiac fibroblasts (CFs), collagen secretion and the underlying mechanism remains unclear. In this study, DyLight® 488 green fluorescent dye or His-tagged proteins were used to label the exogenous serpinE2 protein. It was showed that extracellular serpinE2 translocated into CFs by low-density lipoprotein receptor-related protein 1 (LRP1) and urokinase plasminogen activator receptor (uPAR) of cell membrane through endocytosis. Knockdown of LRP1 or uPAR reduced the level of serpinE2 in CFs and down-regulated the collagen expression. Inhibition of the endocytosis of serpinE2 could inhibit ERK1/2 and β-catenin signaling pathways and subsequently attenuated collagen secretion. Knockdown of serpinE2 attenuates cardiac fibrosis in TAC mouse. We conclude that serpinE2 could be translocated into cardiac fibroblasts due to endocytosis through directly interact with the membrane protein LRP1 and uPAR, and this process activated the ERK1/2, β-catenin signaling pathways, consequently promoting collagen production.     

城市扩张过程中建设用地景观格局演变特征及其驱动力

剖析城市扩张过程中建设用地景观格局演变特征及其驱动力,不仅有助于城市生态环境问题的解决,而且为城市用地结构优化、城市景观规划等工作提供科学依据。以快速城市化的中型城市—扬州市为例,利用多期(1995、2000、2005、2010年和2015年)Landsat卫星影像、乡镇水平的扬州统计年鉴等数据,运用景观格局分析、增强回归树(Boosted regression trees)等方法,研究建设用地的扩张模式、形态及景观格局,定量探究地理、社会和经济因子对景观格局的影响机制,从而明晰景观格局演变特征及其驱动力。结果表明,1995—2015年,建设用地面积持续增加,填充式(Infilling)、边缘式(Edge-expansion)和跳跃式(Leapfrog)3种扩张模式在各时段均有出现,但其优势度随着城市发展而改变。建设用地的形态在城市扩张的过程中经历着"集聚"和"扩散"的交替变化过程,景观格局则出现了同质化倾向,景观破碎化下降、聚合度增加。地理因子(海拔和到县市中心的距离)对景观格局的综合影响虽然高于社会经济,但它的影响力却随着城市发展呈现出下降趋势,社会经济的作用则逐渐增强。海拔和人口...     

内蒙古自治区干旱脆弱性评价

干旱带来的环境影响及经济损失,阻碍了地区的可持续发展。开展干旱脆弱性评价是合理制定区域规划与管理措施的前提条件。然而,国内目前鲜有以省或自治区为研究区域,以市级行政区域为尺度的自然-社会-经济耦合的干旱脆弱性研究。根据IPCC提出的干旱脆弱性评价模型,选取19个指标,在3个维度上(暴露度、敏感度和适应能力)对内蒙古自治区的12个盟市开展了干旱脆弱性评价。采用熵值法确定各指标权重,并用综合指数法和系统分类法计算干旱脆弱性指数并进行分类。研究结果表明,内蒙古自治区的干旱脆弱性呈现由东向西递减的趋势,与干旱脆弱性相关性最强的三个指标分别是第一产业GDP比例、人均可支配收入和第一产业从业人员比例。导致盟市干旱脆弱性的主要贡献因素为人口与人力因素和生态与水资源因素。减缓内蒙古自治区干旱脆弱性可以从加强草原保护建设和管理,合理规划盟市建设,减少人口的集中分布,调整产业结构,提供更多的非农牧就业岗位,加强职业技能培训,完善金融服务和医疗服务等方面入手,从而促进干旱区自然生态和社会经济的可持续发展。     

剑叶龙血树内生放线菌活性菌株的筛选和鉴定

为筛选具有抗菌抗肿瘤活性的药用植物内生菌资源,该文对300余株分离自中国云南西双版纳及越南地区剑叶龙血树的内生放线菌采用琼脂块扩散法进行抗病原细菌活性筛选、平板对峙法进行抗真菌活性筛选、SRB法测定菌株的细胞毒活性及PCR扩增方法筛选非核糖体肽合成酶NRPS基因及聚酮合酶PKS-I、PKS-Ⅱ基因;对得到的优势菌株经8种培养基进行发酵,采用10种病原细菌、3种病原真菌及2种人肿瘤细胞株测试其发酵粗提物的抗菌及抗肿瘤活性以确定最佳发酵培养基;用16S rRNA基因测序方法初步鉴定5株优势菌株的分类地位。结果表明:初筛得到5株抗菌活性、体外细胞毒活性及NRPS、PKS相关基因表达阳性的菌株S01-S05,其中4株(S01-S04)属于链霉菌属、1株(S05)属于类诺卡氏菌属;菌株经不同培养基发酵后产物的抗菌和抗肿瘤活性不同,其中Streptomyces sp. S04在7种培养基中的发酵提取物对8种测试病原菌均有较强抑制作用,且该菌株用Medium C的发酵提取物对人肝癌Hep G2细胞株抑制率达到100%,为剑叶龙血树内生菌活性成分挖掘及新型抗菌药物筛选奠定了基础。     

黑色素母细胞沿神经的迁移

黑色素细胞是皮肤中的特殊细胞,它产生黑色素,并将其传递给周围的角质形成细胞,从而决定皮肤的颜色等,其来源于黑色素母细胞.因此,了解黑色素母细胞如何迁移至靶点,对改善个体肤色以及诊断和治疗黑色素瘤等都具有重要意义.文章通过对黑色素母细胞沿神经迁移这一模式的过程和相关机制进行综述,希望为个体肤色改善以及黑色素瘤术后复发机制...     

sGLP-1对Ⅱ型糖尿病大鼠治疗效果的研究

目的 研究人工合成胰高血糖素样截短肽(sGLP-1)对Ⅱ型糖尿病大鼠的治疗效果.方法 Ⅱ型糖尿病GK大鼠随机分为三组,以合成的GLP-1为阳性对照,观察sGLP-1对Ⅱ型糖尿病GK大鼠血糖水平、胰岛素分泌以及糖耐量的影响,通过MTT法测定sGLP-1对胰岛β细胞系β-TC3增殖作用.结果 与GLP-1相比sGLP-1能够长效控制的血糖水平,明显改善糖尿病大鼠的糖耐量(P<0.01).同时sGLP-1能促进胰岛素分泌和胰岛β-TC3细胞的增殖,使得胰岛体积增大,数量增多.结论 sGLP-1控制血糖的长效能力优于GLP-1,可能从刺激胰岛素分泌和促进胰岛β细胞增殖两个方面对Ⅱ型糖尿病具有治疗作用.     

马铃薯卷叶病毒中国株(PLRV-Ch)复制酶基因结构研究

用设计合成的两对特异性引物,以马铃薯卷叶病毒中国分离株PLRV-Ch RNA为模板,经反转录PCR扩增,将复制酶基因3′端0.6 kb和5′端1.2 kb,克隆于pUC19中,分别构建了重组质粒pLR3和pLR5,并分五段进行了序列分析.将获得的核苷酸序列及氨基酸序列与国外报导的四个PLRV分离株的相应区段的序列同源性进行了比较.结果表明具有高度同源性.文中对核苷酸序列中存在的可能移码序列和其下游的茎环结构或假节结构、以及特征性三次重复区及氨基酸序列中复制酶蛋白N端的碱性氨基酸序列以及C端区域中包括GDD在内的8个特征序列进行了讨论.作者发现移码序列上游的三次重复的核苷酸序列可以形成连续折叠的互补双链区和发夹结构,这一结构可能和转译移码有关.此外PLRV复制酶蛋白N端部分氨基酸序列易变,而C端氨基酸序列十分保守,可能和复制酶功能有更重要关系.