民国初年中国无政府主义自由观剖析

20世纪中国无政府主义者的绝对自由理念,是在对政府、社会、法律的消解过程中建立的,这种建构模式所内含的现实批判因素挤占了学理的创造空间,而无政府主义思想中的卢梭资源又将他们带向终极革命的道路,从追求绝对自由一个极端走向了绝对专制的另一极端,自由成为中国知识分子的乌托邦.     

利用QuantStudioTM 3D数字PCR分析转基因玉米MON863含量

QuantStudioTM 3D数字PCR (QuantStudioTM 3D digital PCR,3D-dPCR)是一种基于超高密度亲疏水微孔芯片实现数字PCR分液原理的新型核酸绝对定量平台,在转基因生物定量领域具有极大的应用前景.本研究基于3D-dPCR平台,以转基因玉米(Zea mays)MON863混合样品为例,建立基于单重和双重数字PCR体系的转基因生物(genetically modified organisms,GMOs)含量分析方法.与传统qRT-PCR比较发现,在缺乏样品纯度、纯合度信息的情况下,数字PCR能够较好地排除这些因素的影响,测定准确的量值.研究结果表明,QuantStudioTM 3D数字PCR是一种适用于转基因生物含量分析的精确定量方法,还可反映转基因玉米种子的基因型.本研究基于3D-dPCR建立的转基因玉米MON863单重和双重定量方法为转基因检测提供了新的方法和参考.     

Analysis of Mycosporine-Like Amino Acids in Selected Algae and Cyanobacteria by Hydrophilic Interaction Liquid Chromatography and a Novel MAA from the Red Alga Catenella repens

Mycosporine-like amino acids (MAAs), a group of small secondary metabolites found in algae, cyanobacteria, lichens and fungi, have become ecologically and pharmacologically relevant because of their pronounced UV-absorbing and photo-protective potential. Their analytical characterization is generally achieved by reversed phase HPLC and the compounds are often quantified based on molar extinction coefficients. As an alternative approach, in our study a fully validated hydrophilic interaction liquid chromatography (HILIC) method is presented. It enables the precise quantification of several analytes with adequate retention times in a single run, and can be coupled directly to MS. Excellent linear correlation coefficients (R² > 0.9991) were obtained, with limit of detection (LOD) values ranging from 0.16 to 0.43 µg/mL. Furthermore, the assay was found to be accurate (recovery rates from 89.8% to 104.1%) and precise (intra-day precision: 5.6%, inter-day precision ≤6.6%). Several algae were assayed for their content of known MAAs like porphyra-334, shinorine, and palythine. Liquid chromatography-mass spectrometry (LC-MS) data indicated a novel compound in some of them, which could be isolated from the marine species Catenella repens and structurally elucidated by nuclear magnetic resonance spectroscopy (NMR) as (E)-3-hydroxy-2-((5-hydroxy-5-(hydroxymethyl)-2-methoxy-3-((2-sulfoethyl)amino)cyclohex-2-en-1-ylidene)amino) propanoic acid, a novel MAA called catenelline.     

Expression Analysis of THBS3 Gene in Different Tissues and Skeletal Muscles at Different Periods from Landrace and Tongcheng Pigs

To study the expression pattern of THBS3 gene in different tissues and during skeletal muscle development, the THBS3 gene expression in different tissues and skeletal muscles during prenatal periods (33, 45, 65, 70 and 90 d) and postnatal periods (0, 9, 30, 60, 120 and 160 d) from Landrace and Tongcheng pigs were detected by Real-time quantification PCR.The results showed that THBS3 gene widely expressed in all tissues examined, exhibiting similar spatial expression patterns with expression peaks in lung in both pig breeds except in stomach and intestine.Moreover, although THBS3 gene showed a significant higher expression level in gestation than after birth in Landrace and Tongcheng pigs (P<0.05), it exhibited different expression patterns between Landrace and Tongcheng pigs, the expression peak was detected at gestation day 45 in Landrace pig, while was detected at gestation day 65 in Tongcheng pig.The results suggested that THBS3 gene involved in skeletal muscle growth and development in pigs, as well as the regulation of asynchronization of skeletal muscle development in different pig breeds.     

计算机图解适线法

该文编制的计算机图解适线法程序,对于总体为皮尔逊Ⅲ型分布的样本系列,能够利用计算机屏幕(或绘图仪)目估适线。主要计算结果为适线法确定的参数X、CV和CS,以及任何指定频率的设计值XP。     

1978—2018年中国农村环境管理政策演进特征——基于206份政策文本的量化分析

基于手工搜集的1978—2018年中国政府颁布的206份农村环境管理政策文本,从政策发布年度、政策发布主体、政策发布形式、政策作用对象、政策工具及政策效力6个维度构建分析框架,运用政策文献计量和内容分析方法分析了中国农村环境管理政策的演进特征。结果表明:1978年改革开放以来,中国农村环境管理政策在数量上呈整体持续上升态势;农村环境管理政策经历了探索起步(1978—1989年)、初步形成(1990—2004年)、快速发展(2005—2013年)和全面推进(2014年以后)4个阶段的演进;政策发布主体的行政级别较高且呈现多元化特征,政策联合决策程度高,但政策之间统筹性和协调性较低;政策绝大多数以通知、意见等形式发出,缺乏权威性和规范性;政策的着力点主要在原生态保护层面,对农村人居环境整治的重视程度不够;命令控制型政策是政府最为偏好的农村环境管理政策工具,经济激励型、信息公开型和自愿参与型政策工具使用不足;政策整体效力的提升主要由政策发布数量驱动,政策反馈不足、政策措施较少和政策目标量化程度低是导致政策平均效力较低的主要原因。     

Development and evaluation of a loop-mediated isothermal amplification assay for rapid and simple detection of Flavobacterium psychrophilum

Flavobacterium psychrophilum is the causative agent of bacterial cold-water disease and rainbow trout fry syndrome of salmonids. The pathogen has been reported from all regions in the world involved in salmonid aquaculture, but also from natural fresh-water environments. We established a quantitative loop-mediated isothermal amplification of DNA (LAMP) method to estimate quantities of F. psychrophilum . LAMP primers were designed based on the sequence of the DNA topoisomerase IV subunit B gene, parE , of F. psychrophilum. parE LAMP exhibited a high specificity for the parE gene of F. psychrophilum but not for other related species. parE LAMP detected the gene in a wide range of concentrations from 2.0 × 10¹ to 2.0 × 10⁹ copies/reaction within 70 min and revealed a good correlation between threshold times and gene copy number.     

Design and validation of a RT‐qPCR procedure for diagnosis and quantification of most types of infectious pancreatic necrosis virus using a single pair of degenerated primers

Infectious pancreatic necrosis virus (IPNV) is an important virus which affects the salmonid aquaculture industry worldwide; therefore, it is important to develop rapid and reliable methods of diagnosis to detect the disease at early stages. Nowadays, RT‐qPCR is replacing other methods because it provides additional information on the viral load, which is important to have a better understanding of the virus replication level and of the stage of the infection and its risk level. The main problem stems from the high diversity of this virus, which can compromise the reliability of the diagnosis. In this study, we have designed an RT‐qPCR procedure for diagnosis and quantification of IPNV based on a single pair of primers targeted to segment B. The procedure has been validated, in vitro and in vivo, testing two different types of standards against seven reference strains and 23 field isolates from different types. The procedure is reliable for the detection of any type, with a detection limit of 31 TCID₅₀ mL^?1, 50 pfu mL^?1 or 66 RNA copies mL^?1, depending on the standard. All the standard curves showed high reliability (R² > 0.95). The results support the high reliability of this new procedure for the diagnosis and quantification of IPNV.     

基于iTRAQ质谱分析技术筛选南方型紫花苜蓿根部响应盐胁迫差异表达蛋白

为了从蛋白质水平上揭示南方型紫花苜蓿(Medicago sativa ‘Millennium’)适应盐胁迫的分子机制,本研究以30 d苗龄的南方型紫花苜蓿实生苗为材料,分析正常培养和250 mmol/L NaCl处理72 h后根中的蛋白表达变化,采用同位素相对标记与绝对定量技术(isobaric tags for relative and absolute quantitation,iTRAQ)结合双向液相色谱与串联质谱(2-dimensional liquid chromatography-tandem mass spectrometry,2D-LC-MS/MS)定量蛋白质组技术鉴定南方型紫花苜蓿根部响应盐胁迫差异表达蛋白,对所获得差异蛋白进行生物信息学分析,筛选出可能耐盐潜在靶标蛋白.同时,选择5个差异表达蛋白进行实时荧光定量PCR(qRT-PCR)验证.结果表明,共鉴定3 857种定量蛋白,534种差异蛋白(变化倍数≥1.2,P＜0.05),其中表达上调的281种,表达下调的253种.基因本体(Gene Ontology,GO)分子功能提示这些差异性蛋白主要参与转运活性、催化活性和酶调控活性.京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)通路显著性富集于代谢途径、次生代谢产物生物合成、苯丙氨酸代谢和核糖体等(P＜ 0.05,错误发现率(false discovery rate,FDR)＜ 0.05).成功鉴定的差异蛋白分别涉及到信号传递(8.99％)、抗氧化物(7.68％)、防御(5.61％)、蛋白质合成、加工和降解(14.79％)、能量产生与转运(5.81％)、代谢(26.97％)、膜与胞内运输(5.62％)和细胞结构、分裂和细胞骨架(2.06％)等.差异表达蛋白中与信号传递、抗氧化物和防御等相关的蛋白表达量总体上调,而与代谢和能量产生与转运相关蛋白表达量总体下调.qRT-PCR实验发现,mRNA与蛋白质表达水平并不一致.研究发现组氨酸磷酸转移蛋白、丝裂原活化蛋白激酶、h类硫氧还蛋白、蛋氨酸亚砜还原酶基因、鸟苷二磷酸(guanosine diphosphate, GDP)解离抑制因子、脂质转移蛋白、β-1,2-木糖基转移酶和H2A/H2B/H3/H4核心组蛋白等可能是紫花苜蓿耐盐潜在靶标蛋白.本研究采用iTRAQ结合2D-LC-MS/MS技术,有效地筛选出南方型紫花苜蓿根部响应盐胁迫差异表达蛋白,为深入认识紫花苜蓿盐胁迫的应答分子调控机制奠定了坚实的基础.     

地表微地形测量及定量化方法研究综述

地表微地形测量是地表粗糙度定量化的基础,对地表形态动态监测、水文过程模拟以及土壤侵蚀过程模型的构建具有重要意义。目前,微地形测量方法主要分为接触式和非接触式2大类,前者包括测针法、链条法、差分GPS法等,后者包括超声波测距法、红外线传感器法、结构光激光扫描法、激光测距扫描法、三维激光扫描仪法、近景摄影测量法等。在全面回顾各方法原理、优缺点及其应用的基础上,分析地表粗糙度定量化常用方法：统计方法指数、地统计学指数和分形及多重分形模型。认为：1）差分GPS法、结构光激光扫描法、三维激光扫描仪法和近景摄影测量法将在亚毫米一,厘米级地表微地形测量及地表粗糙度多尺度特征研究中发挥重要作用,同时简单、方便的测针法可能在野外测量中依然占据主导地位;2）以地表微地形测量技术为基础,在土壤侵蚀过程模型中亟需形成一套完整的“测量一定量化一模型应用”范式,同时应加强对地表微地形空间异质性和各向异性的研究,发展新的统一的地表粗糙度定量化方法。