X-ray Phase Contrast Imaging of Cell Isolation with Super-Paramagnetic Microbeads

Super-paramagnetic microbeads are widely used for cell isolation. Evaluation of the binding affinity of microbeads to cells using optical microscopy has been limited by its small scope. Here, magnetic property of microbeads was first investigated by using synchrotron radiation (SR) in-line x-ray phase contrast imaging (PCI). The cell line mouse LLC (Lewis lung carcinoma) was selected for cell adhesion studies. Targeted microbeads were prepared by attaching anti-VEGFR2 (vascular endothelial growth factor receptor-2) antibody to the shell of the microbeads. The bound microbeads were found to better adhere to LLC cells than unbound ones. PCI dynamically and clearly showed the magnetization and demagnetization of microbeads in PE-50 tube. The cells incubated with different types of microbeads were imaged by PCI, which provided clear and real-time visualization of the cell isolation. Therefore, PCI might be considered as a novel and efficient tool for further cell isolation studies.     

河口地区牡蛎礁的生态功能及恢复措施

在许多温带河口区,牡蛎礁是具有重要生态功能的特殊生境。牡蛎礁在净化水体、提供栖息生境、保护生物多样性和耦合生态系统能量流动等方面均具有重要的功能。近100年来,由于过度采捕、环境污染、病害和生境破坏等原因,许多温带河口区牡蛎种群数量持续下降,河口生态系统的结构与功能受到破坏,富营养化越来越严重。为了修复河口生态系统、净化水质和促进渔业可持续发展,近20年来,世界各地开展了一系列牡蛎礁的恢复活动,尤其美国在东海岸及墨西哥湾建立了大量的人工牡蛎礁,许多研究结果证实,构建的人工牡蛎礁经过2~3年时间,就能恢复自然生境的生态功能。本文介绍了我国首次牡蛎增殖放流工程-长江口南北导堤牡蛎礁,近2年的监测结果显示,长江口导堤牡蛎种群数量快速增长,附近水生生态系统的结构与功能得到明显改善。最后,针对目前牡蛎礁恢复过程中存在的不足,提出了需进一步研究的课题,包括牡蛎基础生物学(病害和分子系统进化),牡蛎礁恢复的关键技术、科学程序及成功的评价标准等。     

分子运动性预测麻栎种子离体胚轴适宜贮藏条件初探

应用电子顺磁共振波谱仪和自旋标记技术,以硝基氧探针为标记物,检测了室温下麻栎种子离体胚轴脱水过程中分子运动性的变化。发现含水量0.7gH2O/gDW至0.64gH2O/gDW范围是细胞质粘度的转折区域,低于这个含水量区域,细胞质粘度骤然上升,推测这个区域是室温下保存离体胚轴的适宜含水量下限。通过变温电子顺磁共振波谱测定,找到离体胚轴含水量在0.43gH2O/gDW至1.02gH2O/gDW范围内,分子运动性的临界温度和玻璃态相变温度所在区间。根据分子运动性随温度变化的规律,预测含水量为0.69gH2O/gDW的麻栎种子离体胚轴适宜贮藏温度约为-50℃。     

不同生境与花部特征对巫山淫羊藿结实特性的影响

对不同生境中巫山淫羊藿结实特性进行比较研究。结果表明:(1)三个种群在繁殖时期中不同花部特征的植株繁殖投资差异不大,如每株分枝数、每株花序数和每株开花数差异均不显著;(2)三个种群中每株结实数、每株结实率、每花序结实数在不同花部特征中差异显著,PLS和SAA要显著大于PESS;(3)三种花部特征在不同生境中单株分枝数和单株花序数均无显著差异。在不同花部特征中种群1开花数量相较少,因而在结实方面均值均要小于种群2和种群3;(4)不同花部特征的果实饱满种子数、败育数和败育率均具有显著差异,而种子数差异不大,但在种群间均无显著的差异。说明种子的多少更多地受到花部特征的影响。总之,巫山淫羊藿的结实特性受到生境和花部特征的双重影响,其中花型比生境对巫山淫羊藿结实特性的影响更为明显。     

螺环哌啶类似物作为DOR激动剂的特征结构研究

阿片受体激动剂与特定阿片受体亚型结合,常用来治疗与外伤、癌症或心脏病相关的严重疼痛,是十分有吸引力的药用物质．阿片受体有3种经典亚型(δ, κ, μ),均有与其对应的激动剂．δ阿片受体(DOR)激动剂因其还有明显的抗焦虑、抗抑郁和器官保护作用,是非常有前景的药物．本文研究了一批共102个N-取代螺环哌啶类似物作为δ阿片受体激动剂的分子,采用比较分子力场(CoMFA)和比较分子相似性指数(CoMSIA)两种分析方法对所有分子进行了三维定量构效关系(3D-QSAR)研究,其中基于疏水场和氢键供体场参数建立的CoMSIA模型最佳,其模型结果为：Q²=0.501,R²_ncv=0.787,R²_pre=0.780,证明模型自我吻合良好,同时有较强的内部及外部预测能力．而模型的等势线图分析表明,在R₁处引入疏水性的取代基及在R₂处引入亲水性的取代基或氢键供体基团对提高激动剂活性有利．这些结论有助于更好地理解N-取代螺环哌啶类似物作为DOR激动剂的机理,为新型的δ阿片受体激动剂的设计和优化提供一定的指导．     

兴安落叶松针叶解剖结构变化及其光合能力对气候变化的适应性

叶片易受环境因子影响,其形态解剖结构特征不但与叶片的生理功能密切相关,而且反映树木对环境变化的响应和适应。叶片结构的改变势必会改变树木的生理功能。同一树种长期生长在异质环境条件下,经过自然选择和适应,会在形态和生理特性等方面产生变异,形成特定的地理种群。另外,母体所经受的环境胁迫也会影响到其子代的生长、发育和生理等特征。因此,了解植物叶片形态结构对环境变化的响应与适应是探索植物对环境变化的响应适应机制的基础。兴安落叶松(Larix gmelinii Rupr.)是我国北方森林的优势树种,主要分布在我国东北地区,但日益加剧的气候变化可能会改变其现有的分布区。为了区分叶片对气候变化的可塑性和适应性,本研究采用同质园法比较测定了6个不同气候条件下的兴安落叶松种源的32年生树木的针叶解剖结构和光合生理相关因子,利用石蜡切片方法分析了针叶的解剖结构特征、光合能力（Pmax-a）、水分利用效率（WUE）之间的关系及其对气候变化的适应性。结果表明：表皮细胞厚度、叶肉细胞厚度、传输组织厚度、维管束厚度、内皮层厚度以及叶片总厚度均存在显著的种源间差异（P < 0.05）。叶肉细胞厚度与Pmax-a、气孔导度和WUE之间均存在显著的正相关关系（P < 0.05）。叶肉细胞厚度、表皮细胞厚度、叶片总厚度以及叶肉细胞厚度和表皮细胞厚度在叶片总厚度中所占比例均与种源地的干燥度指数（即年蒸发量与年降水量之比）呈正线性关系。这些结果说明：不同种源兴安落叶松针叶解剖结构因对种源原地气候条件的长期适应而产生显著的差异,从而引起其针叶光合作用、水分利用等生理功能发生相应的变化,从而有利于该树种在气候变化的情景下得以生存和繁衍。     

Imputing Amino Acid Polymorphisms in Human Leukocyte Antigens

DNA sequence variation within human leukocyte antigen (HLA) genes mediate susceptibility to a wide range of human diseases. The complex genetic structure of the major histocompatibility complex (MHC) makes it difficult, however, to collect genotyping data in large cohorts. Long-range linkage disequilibrium between HLA loci and SNP markers across the major histocompatibility complex (MHC) region offers an alternative approach through imputation to interrogate HLA variation in existing GWAS data sets. Here we describe a computational strategy, SNP2HLA, to impute classical alleles and amino acid polymorphisms at class I (HLA-A, -B, -C) and class II (-DPA1, -DPB1, -DQA1, -DQB1, and -DRB1) loci. To characterize performance of SNP2HLA, we constructed two European ancestry reference panels, one based on data collected in HapMap-CEPH pedigrees (90 individuals) and another based on data collected by the Type 1 Diabetes Genetics Consortium (T1DGC, 5,225 individuals). We imputed HLA alleles in an independent data set from the British 1958 Birth Cohort (N = 918) with gold standard four-digit HLA types and SNPs genotyped using the Affymetrix GeneChip 500 K and Illumina Immunochip microarrays. We demonstrate that the sample size of the reference panel, rather than SNP density of the genotyping platform, is critical to achieve high imputation accuracy. Using the larger T1DGC reference panel, the average accuracy at four-digit resolution is 94.7% using the low-density Affymetrix GeneChip 500 K, and 96.7% using the high-density Illumina Immunochip. For amino acid polymorphisms within HLA genes, we achieve 98.6% and 99.3% accuracy using the Affymetrix GeneChip 500 K and Illumina Immunochip, respectively. Finally, we demonstrate how imputation and association testing at amino acid resolution can facilitate fine-mapping of primary MHC association signals, giving a specific example from type 1 diabetes.     

Mutation Inactivation of Nijmegen Breakage Syndrome Gene (NBS1) in Hepatocellular Carcinoma and Intrahepatic Cholangiocarcinoma

Nijmegen breakage syndrome (NBS) with NBS1 germ-line mutation is a human autosomal recessive disease characterized by genomic instability and enhanced cancer predisposition. The NBS1 gene codes for a protein, Nbs1(p95/Nibrin), involved in the processing/repair of DNA double-strand breaks. Hepatocellular carcinoma (HCC) is a complex and heterogeneous tumor with several genomic alterations. Recent studies have shown that heterozygous NBS1 mice exhibited a higher incidence of HCC than did wild-type mice. The objective of the present study is to assess whether NBS1 mutations play a role in the pathogenesis of human primary liver cancer, including HBV-associated HCC and intrahepatic cholangiocarcinoma (ICC). Eight missense NBS1 mutations were identified in six of 64 (9.4%) HCCs and two of 18 (11.1%) ICCs, whereas only one synonymous mutation was found in 89 control cases of cirrhosis and chronic hepatitis B. Analysis of the functional consequences of the identified NBS1 mutations in Mre11-binding domain showed loss of nuclear localization of Nbs1 partner Mre11, one of the hallmarks for Nbs1 deficiency, in one HCC and two ICCs with NBS1 mutations. Moreover, seven of the eight tumors with NBS1 mutations had at least one genetic alteration in the TP53 pathway, including TP53 mutation, MDM2 amplification, p14ARF homozygous deletion and promoter methylation, implying a synergistic effect of Nbs1 disruption and p53 inactivation. Our findings provide novel insight on the molecular pathogenesis of primary liver cancer characterized by mutation inactivation of NBS1, a DNA repair associated gene.     

神经元形态重建工具的研究进展

近年来,分子标记和显微光学成像技术的系列突破,使得单细胞分辨的全脑尺度神经群落成像成为现实.然而,现有神经元形态重建工具的发展速度远远滞后于海量数据的产生速度,难以满足现阶段成像数据的分析需求.在此背景下,我们首先分析了神经元形态重建工具发展滞后的原因,简述现有半自动和全自动神经元形态重建工具的特点和最新发展,并结合现有工具的特点分析其向高通量、高准确度重建工具发展时面临的挑战.最后,我们对未来形态重建工具的发展趋势及应用前景做出展望.