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。     

新城疫病毒V4株L基因的克隆与序列分析

设计了 2对引物V4L1、V4L2和V4L3、V4L4 ,用RT PCR法对新城疫病毒 (NDV )V4克隆株的L基因进行了分段克隆。用V4L1、V4L2扩增出约 4 .1kb的V 4LA片段 ,用V4L3、V4L4扩增出约 3.5kb的V 4LB片段 ,分别对克隆出的 2个片段进行序列测定 ,并用DNAsis软件比较分析后进行拼接 ,得到长约 7.2kb、包含有NDVV4克隆株L基因全长的核苷酸序列。NDVV4克隆株L基因mRNA全长为 6 70 4bp ,拥有 1个 6 6 15bp的开放阅读框 ,推测其编码的氨基酸数为 22 0 4个。氨基酸同源性分析表明 :V4克隆株与HB92V4株L基因的同源性为 99.0 % ,与LaSota、B1、B1T(美国Takaaki分离株 )、BeaudetteC、Clone30、F4 8E9、SF0 2和ZJ1株的同源性为 94 .1%～96 .5 %。     

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。     

伪狂犬病病毒闽A株糖蛋白gD基因去信号肽片段在大肠埃希氏菌中的表达

应用PCR方法从包含伪狂犬病病毒(PRV)Fa株糖蛋白gD基因的重组质粒pB13中扩增出糖蛋白gD基因去信号肽片段,将其克隆到大肠埃希氏菌表达载体pThiohisA中。测序结果表明,糖蛋白gD基因去信号肽片段长1 155 bp,与PRV Fa株糖蛋白gD基因完全一致。经1mmol/L IPTG诱导后,重组质粒pThiohisA-gD在大肠埃希氏菌XL1-blue、Top10、DE3和DH5α中均得到表达,其中在宿主菌XL1-blue中表达量最高。Western-blotting结果显示,大肠埃希氏菌XL1-blue表达的糖蛋白gD具有免疫原性。     

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.     

美国冒犯性商标注册审查标准的变化及其启示——从涉新冠疫情商标申请谈起

《美国兰哈姆法》第2条a款和我国《商标法》第10条第1款第8项在国际公约义务和惯例、立法目的、条文开放性上有相似之处,但在商标注册层面的表达自由和遵循公序良俗边界的认定上差异巨大。表达自由不应逾越遵循公序良俗的国际惯例和国内司法习惯,美国式的完全放开商标注册层面的表达自由不符合我国传统价值观下公众的合理预期。我国在适用"不良影响"条款时需要把握表达自由与遵循公序良俗二者的界限,避免过宽适用"不良影响"条款而吞噬表达自由的空间,应以本土性价值判断克制条款的适用。     

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.     

The Legal Protection for China's Flexible Workers during the Covid-19 Pandemic

The COVID-19 pandemic has caused a chain of socioeconomic effects worldwide. China’s social and economic operations and labor market have also been suffered because of this global crisis. During hard times, flexible employment becomes an important means for stabilizing employment. However, the pandemic also exposed the problems ensuing from insufficient legal protection for flexible workers. First, there are some gaps and defects in the labor and social security laws in terms of their scope and applicability to flexible workers. Second, there were limitations in the support and protection policies for flexible workers at the early stages of the pandemic. Although the government implemented measures soon after the outbreak of COVID-19, their role is still constrained by the legal lacunae in protecting flexible workers. As flexible employment will continue to play a significant role in China’s labor market, the State Council issued a far-reaching official document in July 28, 2020 to support it. This document indicates the directions for providing better legal protection for flexible workers, which contains expanding the legal scope and enriching the legal protection. Based on the realities and latest policies, the legal protection for flexible workers should be systemically improved in the future.