微小牛蜱幼蜱cDNA表达文库的构建

为了研究微小牛蜱功能性抗原基因,从微小牛蜱(Boophilus microplus)幼蜱中提取总RNA并分离纯化mRNA。采用Oligo(dT)引物合成双链cDNA,在其两端加EcoRⅠ、HindⅢ定向接头,用Mini Column Fractionation凝胶过滤柱纯化后定向克隆到λSCREEN载体。经体外包装,成功构建了我国微小牛蜱幼蜱cDNA表达文库,铺板测定原始库容量为4.5×105PFU,扩增后的滴度为7.2×1010PFU/mL。     

猪传染性胃肠炎病毒TH-98株M蛋白基因的克隆及原核表达

构建了TGEV TH-98株M基因序列的原核表达载体,利用RT-PCR方法从TGEVTH-98株中扩增出M蛋白基因,并亚克隆至原核表达载体pGEX-6P-1上,转化大肠埃希氏菌BL21(DE3)pLysS,用IPTG诱导重组菌株表达了融合蛋白。结果表明,从TGE TH-98株克隆到了M基因cDNA,该基因全长789 bp,与T014株、Purdue株、TFI株和96-1933株的M基因同源性分别为92.37%、98.35%、99.87%和96.96%。SDS-PAGE电泳结果显示,重组表达质粒获得了约57 ku的目的带,其中M蛋白的分子质量约为31 ku,与预期的结果一致;Western-blotting分析表明,融合表达产物可与TGEV全病毒制备的阳性血清发生反应。证实,TGEV的M基因高度保守,构建的M基因原核表达载体pGEX-6P-TM能在大肠埃希氏菌中获得高效表达。     

猪传染性胃肠炎病毒S基因重组腺病毒的构建及特性研究

将克隆的猪传染性胃肠炎病毒(TGEV)全长S基因插入pAdTrack-CMV穿梭质粒中,形成的转移质粒pAdTrack-CMV/S线性化后,用pAdEasyTM系统电转化含有腺病毒骨架载体pAdEasy-1的感受态大肠埃希氏菌BJ5183,经同源重组,构建了含有全S基因的重组腺病毒载体,经酶切充分暴露反向末端重复序列后,与脂质体混合转染AD-293细胞,成功获得了AD-S复制缺陷型腺病毒,经检测证实,AD-S稳定性好,安全性高。转录水平检测证实,S基因得到了转录;荧光检测发现,外源蛋白已得到表达。     

H5N1亚型禽流感病毒NA基因在重组禽痘病毒中的表达及其产物的免疫原性

将禽流感病毒A/Goose/Guangdong/1/96(H5N1)株的NA基因及LacZ报告基因克隆到禽痘病毒载体pSY681中,构建重组转移载体pSY(NA+LacZ),将其转染鸡胚成纤维细胞,经蓝白斑筛选,纯化表达NA基因的重组禽痘病毒rFPV-NA,用其免疫SPF鸡,检测该重组禽痘病毒的免疫原性。结果显示,该重组禽痘病毒能够在体外培养细胞内表达具有生物学活性的NA蛋白,表达产物的分子质量约为55 ku;用该重组禽痘病毒免疫SPF鸡后,能够使免疫鸡获得对H5N1和H7N1两种亚型HPAIV攻击的部分保护。表明,重组禽痘病毒rFPV-NA具有良好的免疫原性。     

Automated Classification of Epiphyses in the Distal Radius and Ulna using a Support Vector Machine

The aim of this study was to automatically classify epiphyses in the distal radius and ulna using a support vector machine (SVM) and to examine the accuracy of the epiphyseal growth grades generated by the support vector machine. X‐ray images of distal radii and ulnae were collected from 140 Chinese teenagers aged between 11.0 and 19.0 years. Epiphyseal growth of the two elements was classified into five grades. Features of each element were extracted using a histogram of oriented gradient (HOG), and models were established using support vector classification (SVC). The prediction results and the validity of the models were evaluated with a cross‐validation test and independent test for accuracy (P_A). Our findings suggest that this new technique for epiphyseal classification was successful and that an automated technique using an SVM is reliable and feasible, with a relative high accuracy for the models.     

Classification of firing pin impressions using HOG-SVM

Crimes, such as robbery and murder, often involve firearms. In order to assist with the investigation into the crime, firearm examiners are asked to determine whether cartridge cases found at a crime scene had been fired from a suspect's firearm. This examination is based on a comparison of the marks left on the surfaces of cartridge cases. Firing pin impressions can be one of the most commonly used of these marks. In this study, a total of nine Ruger model 10/22 semiautomatic rifles were used. Fifty cartridges were fired from each rifle. The cartridge cases were collected, and each firing pin impression was then cast and photographed using a comparison microscope. In this paper, we will describe how one may use a computer vision algorithm, the Histogram of Orientated Gradient (HOG), and a machine learning method, Support Vector Machines (SVMs), to classify images of firing pin impressions. Our method achieved a reasonably high accuracy at 93%. This can be used to associate a firearm with a cartridge case recovered from a scene. We also compared our method with other feature extraction algorithms. The comparison results showed that the HOG-SVM method had the highest performance in this classification task.     

论同步录音录像的口供功能

实践中,同步录音录像多用作视听资料,证明讯问笔录的合法性,从而忽略了它由于记录了口供及其非文字信息而具有的作为口供证据的功能。同步录音录像的口供功能包括通过查看同步录音录像,发现案件线索,找到案件侦查突破口,也包括作为口供证据直接证明犯罪事实。检察机关将同步录音录像作为口供使用可以更好地解决检察机关自侦案件证据缺乏的难题。为实现上述同步录音录像的口供功能,应当提高侦查人员分析判断同步录音录像的能力,积极将同步录音录像作为供述证据使用,避免法官在查看同步录音录像时的认知偏见等等。     

少数民族地区网络舆情危机诉求表达及其参与主体的特殊性分析

相对于一般的网络舆情主体之诉求指向,少数民族公众在网络舆情危机中体现的意愿、诉求、指向等态度与情绪,大都集中于利益表达、社会抗议、行为动员三个方面。此外,少数民族地区网络舆情危机中的主体特殊性主要表现为:主体间的"数字鸿沟"现象更为明显;参与主体成分的多元性特征十分突出。     

侵犯著作权的惩罚性赔偿与表达自由

著作权侵权具有易发性、难以预防和控制、侵权成本低与维权成本高的特点。传统民法认为,侵权损害赔偿只具有填补损害的功能。然而,作品作为一种典型的公共产品,著作权人对作品的控制较弱,补偿性损害赔偿难以为著作权提供足够保护。因此,在著作权侵权救济中应引入惩罚性赔偿。同时,考虑到惩罚性赔偿有可能会不合理地限制作品的传播进而影响到社会公众的表达自由,在惩罚性赔偿的适用范围和条件上应当做出一定限制。     

Unblocking the Digital Economy Act 2010; human rights issues in the UK

Through an example of a study utilizing the case-law research method, this paper critically assesses whether taking into account both the findings of Mr La Rue (the United Nations Rapporteur on Human Rights) as well as some Court of Justice of the European Union's (CJEU) case-law, website blocking could be implemented in a way which is compatible with the European Convention on Human Rights (ECHR), in particular, with Article 10. Drawing upon, inter alia, Ofcom site blocking review, sections 17 and 18 of the Digital Economy Act 2010 (DEA), section 97A of the Copyright, Designs and Patents Act 1988 (CDPA), and some independent expert evidence, this paper's major argument is that in view of the CJEU SABAM v Scarlet and SABAM v Netlog, the UK government's decision to repeal the website blocking provisions of the DEA appears appropriate. The paper examines the findings of Fox v BT. It contrasts such findings with the CJEU's case-law and in light of the incompatibility of any website blocking measure with the cumulative three-part test set out in the United Nations Rapporteur on Human Rights discusses a number of implications. It concludes that given that the implementation of content blocking systems, such as Cleanfeed is likely to result in general monitoring being carried out; the UK government could possibly be in breach of EU law, namely, Article 15(1) of Directive 2000/31.