鸡传染性腔上囊病病毒Gt株全基因组序列分析及真核表达载体的构建

采用一步法长距离RT-PCR克隆了IBDV Gt株全基因组,对其核苷酸和氨基酸序列进行了系统分析。在IBDV Gt株全基因组中引入分子标签,且在基因组两端分别引入锤头状核酶结构(HamRz)和丁肝病毒核酶结构(HdvRz)。将带有分子标签和核酶结构的IBDV基因组插入载体pCAGGS的β肌动蛋白启动子下游,构建了IBDV真核表达载体pCAGGmGtAHRT和pCAG-GmGtBHRT,为IBDV新型高效拯救平台的构建及基于此的病毒基因功能奠定了基础。     

Venezuela,April 2002: Coup or Popular Rebellion? The Myth of a United Venezuela

This article assesses the merits of opposing National Assembly reports into the coup against President Chávez of Venezuela in April 2002. Looking at the historical context and the content of the reports, it argues that the two opposing accounts reflect a class division that has always existed in Venezuela but has been officially denied. It concludes that a possible exit from the stalemate could be that the opposition accept the reality of this class division and therefore the Chávez government as a legitimate representative of the popular classes. This, however, is unlikely in the present circumstances.     

毒鼠强诱导细胞DNA损伤的彗星电泳检测

目的　研究毒鼠强对小鼠淋巴细胞和脑细胞DNA的损伤作用。方法　分离健康小鼠的淋巴细胞和脑细胞 ,以彗星电泳的方法测定不同浓度毒鼠强处理后的细胞DNA损伤。结果　 1/2 0～ 1/2LD50 剂量组的毒鼠强均可引起淋巴细胞和脑细胞不同程度的DNA损伤 ,与对照组呈极显著性差异 (P <0 0 0 1)。结论　毒鼠强引起细胞DNA断裂损伤 ,并呈现明显的剂量 -效应关系。     

6FAM-HEX-TMR-ROX四色荧光分析体系的构建

目的为在310基因分析仪上进行6FAM-HEX-TMR-ROX四色荧光STR复合扩增分型建立基础条件。方法以pGEM载体作为靶DNA,设计引物扩增获得500~80bp的13个长度不等DNA片段,以这些片段纯化物作为模板DNA,分别扩增获得6FAM、HEX、TMR和ROX标记的M atrix标准物,用4种M atrix标准物在310基因分析仪上构建可用于6FAM-HEX-TMR-ROX四色荧光分析的M atrix文件。结果扩增所得的13个非标记片段,长度分别为80、100、120、140、160、180、200、220、260、300、340、400、500bp,在非变性聚丙烯酰胺凝胶连续电泳-银染凝胶上均表现为单条带,6FAM、HEX、TMR、ROX分别标记的片段通过310基因分析仪检测均为单峰。混合ROX标记的80、120、180、220、260、300bp的6个片段获得的内标,显示出良好的线性关系。结论建立了有效的6FAM-HEX-TMR-ROX四色荧光分析M atrix,为进行多个STR基因座荧光标记复合扩增检测奠定了基础。     

驯鹿狂蝇生活习性的观察

对感染驯鹿狂蝇幼虫驯鹿进行了剖检观察,证实狂蝇幼虫主要寄生于驯鹿鼻腔及咽喉黏膜,在当年的7月到翌年1月主要以一期幼虫为主,从翌年2～3月开始快速生长发育,在4～6月多为二、三期幼虫。从翌年5月开始,成熟的三期幼虫随驯鹿打喷嚏排出体外羽化为成蝇。通过对驯鹿狂蝇生活史的观察,表明在大兴安岭地区,驯鹿狂蝇三期幼虫在50％～70％的湿度、21．5℃的平均温度及21．5℃下的451．5～666．5℃总积温下,经21-31 d羽化为成蝇,羽化率为59．6％。成蝇出现于6-9月份,在7～8月份达高峰期。驯鹿狂蝇1年发生一个世代,在驯鹿体内的寄生生活长达9-10个月;雌性成蝇每次产胎蚴30-80条,一生共产800-1000条;雌蝇寿命比雄蝇长,约为1-3周。     

珍灵口服液的主要药效与毒性研究

目的：对珍灵口服液进行主要药效及毒性研究。方法：采用酶联免疫吸附（ELISA）测定小鼠血清中IgG含量，采用植物血凝素（PHA）刺激淋巴细胞转化－－小鼠体内诱导法测定淋巴细胞转化率；用TBA反应比色法测定小鼠血清中脂质过氧化物（LPO）的含量，用NBT法（四氮唑蓝法）测定超氧化物歧化酶（SOD）的含量；并进行了急性和长期毒性试验。结果：小鼠血清IgG含量、淋巴细胞数、红细胞内SOD含量明显升高（P＜0．05或P＜0．01）；血清中LPO含量明显降低（P＜0．05）；未见毒性反应。结论：珍灵口服液能增强机体免疫力，并具有降低机体损伤的功能，是安全有效的保健药品。     

口蹄疫病毒多基因片段的克隆及其杆状病毒表达载体的构建

根据定点突变原理获得了包含口蹄疫病毒P1、2A、3C、3D及部分 2B编码区的目的基因片段 ,经SpeⅠ和HindⅢ双酶切后 ,与经相同方法处理的杆状病毒转移载体 pFastBacHT连接 ,得到了重组质粒 pFB P12X3C3D。经酶切和测序鉴定后 ,将其转化入含穿梭载体Bacmid的感受态细胞DH10Bac ,经抗性及蓝白斑筛选 ,得到了含P12X3C3D多基因的重组穿梭载体 ,将其命名为Bacmid P12X3C3D ;提取其DNA并转染Sf9昆虫细胞 ,最后获得含口蹄疫病毒P12X3C3D多基因的重组杆状病毒。     

退耕还林保护西部生态环境

生态环境的恢复与建设是我国西部大开发的首要任务.在保护生态、改善环境、发展经济的过程中,要特别重视退耕还林(草)等对保护西部生态环境的作用.我们要根据<全国生态环境建设规划>以及可持续发展的要求,采取相应的对策.     

全国人大常委会在宪法实施中的创制行为及其界限——以“辽宁贿选案”为例

“辽宁贿选案”发生后,该省人大常委会无法正常开会履职。对此,全国人大常委会决定成立辽宁省十二届人大七次会议筹备组,代行该省人大常委会的部分职权。这被认为是宪法上的创制性安排。全国人大常委会的创制超出了宪法文本,不是宪法解释行为,也不是宪法续造行为,而是宪法建造行为。政治主体在面对宪法僵局时主动实施宪法,建造出民主集中制原则的具体内容和人民代表大会制度的新内涵,有效解决僵局并发展宪法。这并非孤例。全国人大常委会还曾在“总理辞职案”中进行过创制。创制需要符合一定的条件,前提是出现了必须创制的环境、没有可直接适用的宪法规则,方法是按照法定程序作出政治选择,形式是发布决定而非立法。创制具有宪法法律界限和合理性界限。政治主体要选择恰当的宪法原则和制度进行创制,所创制的内容要符合比例原则。创制是全国人大常委会掌握的一种新宪法工具,表明我国宪法实施具有政治主体与宪法互动的特征。     

DNA-based and taxonomic identification of forensically important Sarcophagidae (Diptera) in southeastern Spain

Studying dipterans at the scene of a death can provide essential information for interpreting the evidence and help to reconstruct the events happened to a corpse in the past. Molecular tools have been employed for identification at specific levels in the cases of cryptic species or poorly conserved specimens. Identification of specimens is essential in forensic entomology since each species has a specific growth rate, which determines the calculation of the minimum post mortem interval (minPMI). In addition, phylogeographic reconstruction within a species can help to differentiate the haplotypes from a geographic area, thereby helping to clarify the possible relocation of a corpse. The morphological identification of Sarcophagidae species is often difficult, especially for the females. This is an important Diptera family since some of its species are among the first to reach a corpse, especially in warm areas. In this study, we compared the sarcophagids found in human corpses in forensic cases in Alicante (southeast of Spain) with specimens collected from baited traps in the same area and surrounding provinces. In total, 189 specimens were collected, comprising 72 from forensic cases and 117 from baited traps. Molecular identification was conducted by sequencing the cox1 mitochondrial gene and analyzing the sequences using ABGD, GMYC, and BIN species delimitation methods. The median joining algorithm in the PopART program was used to construct phylogeographic networks. Eight species in the family Sarcophagidae were identified. The most widely collected species were Sarcophaga argyrostoma and Sarcophaga tibialis. The haplotype networks obtained for these species did not indicate a clear geographic distribution of haplotypes. The S. argyrostoma samples from Alcoy were clearly isolated. The results demonstrated that this method is useful for identifying Sarcophagidae samples in forensic investigations and it can be employed for minPMI estimation.