Microtubules that form the stationary lattice of muscle fibers are dynamic and nucleated at Golgi elements

Skeletal muscle microtubules (MTs) form a nonclassic grid-like network, which has so far been documented in static images only. We have now observed and analyzed dynamics of GFP constructs of MT and Golgi markers in single live fibers and in the whole mouse muscle in vivo. Using confocal, intravital, and superresolution microscopy, we find that muscle MTs are dynamic, growing at the typical speed of ∼9 µm/min, and forming small bundles that build a durable network. We also show that static Golgi elements, associated with the MT-organizing center proteins γ-tubulin and pericentrin, are major sites of muscle MT nucleation, in addition to the previously identified sites (i.e., nuclear membranes). These data give us a framework for understanding how muscle MTs organize and how they contribute to the pathology of muscle diseases such as Duchenne muscular dystrophy.     

Specificity of miR-378a-5p targeting rodent fibronectin

One criterion for microRNA identification is based on their conservation across species, and prediction of miRNA targets by empirical approaches using computational analysis relies on the presence of conservative mRNA 3′UTR. Because most miRNA target sites identified are highly conserved across different species, it is not clear whether miRNA targeting is species-specific. To predict miRNA targeting, we aligned all available fibronectin 3′UTRs and observed significant conservation of all 20 species. Twelve miRNAs were predicted to target most fibronectin 3′UTRs, but rodent fibronectin showed potential binding sites specific for five different miRNAs. One of them, the miR-378a-5p, contained a complete matching seed-region for all rodent fibronectin, which could not be found in any other species. We designed experiments to test whether the species-specific targeting possessed biological function and found that expression of miR-378a-5p decreased cancer cell proliferation, migration, and invasion, resulting in inhibition of tumor growth. Silencing fibronectin expression produced similar effects as miR-378a-5p, while transfection with a construct targeting miR-378-5p produced opposite results. Tumor formation assay showed that enhanced expression of fibronectin in the stromal tissues as a background environment suppressed tumor growth, while increased fibronectin expression inside the tumor cells promoted tumor growth. This was likely due to the different signaling direction, either inside-out or outside-in signal. Our results demonstrated that species-specific targeting by miRNA could also exert functional effects. Thus, one layer of regulation has been added to the complex network of miRNA signaling.     

对叶榕榕果内生细菌分离鉴定及抑菌活性的研究

【目的】为植物-内生细菌的生态关系研究, 以及植物内生细菌资源的利用提供一定的依据。【方法】采用微生物学传统分离培养的方法从对叶榕果实中分离到内生细菌54株, 通过限制性酶切分析(ARDRA)共有16个操作分类单元(Operational taxonomic units, OTUs), 对16株代表菌株16S rDNA序列系统发育分析。【结果】16个菌株都找到了与其相似性最高的菌株, 相似性达到95%?100%。其中6株为芽孢杆菌Bacillus属, 为对叶榕果实内生细菌优势菌属; 3株为Staphylococcus属, 2株为Pseudomonas属, 1株为Serrata属, 1株为非培养细菌的同源菌, 1株为Kocuria属, 1株为Delftia属, 还有1株为Acinetobacter属。【结论】这16株内生菌在系统发育树中明显聚为两大支; 在参与抑菌试验的14株内生菌中, 有13株对受试菌有不同程度的拮抗作用。尤其是其中的芽孢菌属的Swx15和不动杆菌属的Swx25菌株, 抑菌作用较强, 且有较广的抑菌谱性。     

基于EMA-qPCR的茄科青枯菌活体检测技术的建立

【目的】利用特异性核酸染料叠氮溴乙锭(Ethidium monoazide bromide, EMA)与实时荧光定量PCR技术相结合, 建立一种能有效区分青枯菌死活细胞的检测方法。【方法】样品DNA制备前经EMA渗透预处理, 再进行实时荧光定量PCR特异扩增菌体DNA。【结果】终浓度为2.0 mg/L的EMA能有效排除1.0×107 CFU/mL灭活青枯菌细胞DNA的扩增, 对活细胞和不可培养状态(Viable but non-culturable, VBNC)活菌的DNA扩增均没有影响。当每个定量PCR反应体系中的活细胞在5.0×100?5.0×104 CFU范围内时, 扩增Ct值与定量PCR反应体系中活细胞CFU对数值呈良好的负相关性(R2=0.992 5)。比较EMA-qPCR法和平板计数法对经过不同温度短期保存的青枯菌检测结果发现, 待检样品可在24 °C与4 °C冷藏条件下短期保存。【结论】本研究建立的EMA-qPCR方法能有效检测青枯菌VBNC细胞和有效区分死活菌, 避免或减少青枯菌PCR检测的假阳性和假阴性。     

Coxsackievirus A16 infection triggers apoptosis in RD cells by inducing ER stress

Coxsackievirus A16 (CA16) infection, which is responsible for hand, foot and mouth disease (HFMD), has become a common health problem in Asia due to the prevalence of the virus. Thus, it is important to understand the pathogenesis of CA16 infection. Viruses that induce endoplasmic reticulum (ER) stress are confronted with the unfolded protein response (UPR), which may lead to apoptotic cell death and influence viral replication. In this study, we found that CA16 infection could induce apoptosis and ER stress in RD cells. Interestingly, apoptosis via the activation of caspase-3, -8 and -9 in the extrinsic or intrinsic apoptotic pathways in RD cells was inhibited by 4-phenyl butyric acid (4PBA), a chemical chaperone that reduces ER stress. These results suggest that CA16 infection leads to ER stress, which in turn results in prolonged ER stress-induced apoptosis. This study provides a new basis for understanding CA16 infection and host responses.     

1961-2005年东北地区气温和降水变化趋势

利用东北地区96个气象站1961-2005年的日平均气温和日降水量资料,采用线性倾向率法,累积距平法,Mann-Kendall法和Morlet小波分析等方法,对东北地区近45年来的气候变化和突变现象进行了研究.结果表明:近45年来,东北地区年平均气温变化在2.45-5.72℃之间,年均温呈现显著上升趋势,气候倾向率为0.38 ℃/10 a(P＜0.01),1988-1989年间发生了由低温到高温的突变;东北地区四季平均气温均呈现增高的趋势,其中冬季气温增幅最大,气候倾向率达到0.53℃/10 a,夏季气温增幅最小,0.24 ℃/10 a.东北地区年均温和季节均温年代际变化亦呈现明显的增暖趋势,年均温,春季均温和冬季均温均在1981-1990年开始变暖,夏季均温和秋季均温在1991-2000年开始变暖.东北地区气温增暖幅度随纬度的升高而增大,大兴安岭北部和小兴安岭地区是增温最明显的地区,增暖幅度较小的地区为辽河平原,辽东半岛和长白山南部地区.东北地区年降水量变化在430.40-678.72 mm之间,降水量变化趋势不明显,整体呈现减少趋势,气候倾向率为-5.71 mm/10 a(P＞0.05),1981-1990年为降水最多的年代,1982-1983年间发生了降水量由少到多的突变.四季降水量变化呈现不同的趋势,其中春季和冬季降水量呈现增多的趋势,夏季和秋季降水量呈现减少的趋势.降水量减少较明显的地区为辽东半岛和长白山南段,降水量增多较明显的地区为大兴安岭北部和松嫩平原.Morlet小波分析结果表明,东北地区年平均气温存在11a的强显著周期,此外还有24 a和6a尺度的变化周期;东北地区年降水量存在16a的强显著周期和6 a的小尺度变化周期.通过以上分析,近45 a东北地区总体气候呈现明显暖干化趋势.     

气候变化对东北沼泽湿地潜在分布的影响

东北地区是我国沼泽湿地分布最广泛的地区。为研究沼泽湿地对气候变化的响应,选取了对沼泽湿地分布可能存在影响的26个环境因子,利用最大熵(Maximum Entropy, MaxEnt)模型模拟了沼泽湿地基准气候条件下的潜在分布,并预测了气候变化情景下2011-2040 年、2041-2070 年和2071-2100 年3个研究阶段东北沼泽湿地潜在分布。研究结果表明:最大熵模型预测精度较高(平均AUC(Aera Under Curve)为(0.826±0.005))。基准气候条件下东北沼泽潜在分布区主要为大小兴安岭和三江平原地区。随着时间的推进,东北地区沼泽湿地原有潜在分布面积明显减少,而新增潜在分布面积较少,总面积呈现急剧减少趋势。至2071-2100年,原有沼泽湿地潜在分布面积将减少99.80%,新增潜在分布面积仅2.48%,总潜在分布面积减少97.32%。空间分布上,东北沼泽湿地潜在分布呈现由东向西迁移,南北向中心收缩的趋势。研究结果可为东北地区沼泽湿地保护政策的制定提供参考。     

中国大陆归化植物新记录

报道了中国大陆归化植物新记录刺毛峨参(Anthriscus caucalis M.Bieb.)和印加孔雀草(Tagetes minuta L.),并进行了形态描述,提供了凭证标本和活植物照片。     

罗格列酮对AD样小鼠学习记忆减退的影响及机制

研究了罗格列酮对链脲佐菌素(streptozotocin, STZ)脑室内注射的阿尔茨海默病(AD)模型小鼠学习记忆减退的影响及机制．在小鼠脑室内注射STZ建立AD模型,治疗组小鼠采用罗格列酮灌胃给药30天．Morris水迷宫实验检测小鼠学习记忆能力,免疫印迹和免疫荧光检测Tau蛋白的磷酸化、神经丝(NFs)蛋白的磷酸化及糖基化、JNK和ERK蛋白的表达,微管结合实验检测Tau蛋白与微管的组装功能,荧光染料Fluoro-Jade B检测小鼠脑内退变神经元．结果显示,相比对照组,模型组小鼠平均逃避潜伏期和路径长度明显增加、穿越隐匿平台次数明显减少、Tau和NFs蛋白表达过度磷酸化、NFs蛋白糖基化减弱,而用罗格列酮干预的小鼠学习记忆改善并且Tau和NFs蛋白的磷酸化水平降低、NFs蛋白糖基化水平增加,Tau蛋白与微管结合能力改善,模型组JNK的磷酸化高于对照组和治疗组、模型组ERK1的磷酸化低于对照组和治疗组、各组在ERK2磷酸化无明显差异,模型组小鼠脑中FJB标记的退化神经元明显多于对照组和治疗组．结果说明,罗格列酮能改善STZ脑室内注射引起的小鼠学习记忆减退,其机制可能与改善胰岛素信号通路、降低Tau和NFs蛋白的过度磷酸化、减少神经退行性变有关．     

染色质免疫沉淀-测序：全基因组范围研究蛋白质-DNA相互作用的新技术

染色质免疫沉淀-测序(ChIP-seq)是近年来新兴的将染色质免疫沉淀(ChIP)与深度测序技术相结合,在全基因组范围内分析DNA结合蛋白结合位点、组蛋白修饰、核小体定位和DNA甲基化的高通量技术．在新一代测序(NGS)技术的大力推动下,ChIP-seq提供了一种相对于ChIP-chip高分辨率、低噪音、高覆盖率的研究方法．随着测序成本的降低,ChIP-seq逐步成为研究基因调控和表观遗传机制的一种常用手段．本文就该技术的最近研究进展进行综述,并着重介绍ChIP-seq数据分析过程及该技术的实际应用情况.