2008～2009年浙江省猪圆环病毒2型全基因组序列的比较和分析

应用PCR方法对2008～2009年采集的99份疑似断奶仔猪多系统衰竭综合征病猪的脾、肺、淋巴结混合组织样本进行猪圆环病毒2型(PCV2)检测,并以组织DNA为模板进行了PCV2的全基因组扩增,克隆后进行了测序。结果显示,2008年与2009年浙江省PCV2的平均阳性率为47.5%,与2004～2006年相比基本持平。随机选取18份阳性病料进行PCV2全基因组测序分析,结果,18个毒株均属于Group1,其中5个毒株(27.8%)属于1A分支,3个毒株(16.7%)属于1B分支,9个毒株(50%)属于1C分支。表明,Group1依然是浙江省PCV2的主导基因型,且1C分支从无到有,并逐渐占据主导地位。     

猪生殖与呼吸综合征病毒宁夏分离株ORF5基因的克隆表达及其表达产物的活性分析

参考已发表的猪生殖与呼吸综合征病毒(PRRSV)ORF5基因序列,设计合成了1对覆盖完整ORF5基因区段的引物,克隆了PRRSV NX/HY株的ORF5基因(登录号为EU600287),并进行了序列比较.将该基因克隆到原核表达栽体pGEX-6P-1中,构建了融合表达质粒pGEX-6P-1-5并转化BL21,对表达蛋白进行Western-blot分析.结果表明,NX/HY分离株ORF5基因与PRRSV JX1、CH-la株的核苷酸序列同源性分别为99.2%和95.2%,推导的氨基酸序列的同源性分别为98.5%和92.5%;层析扫描分析显示,目的蛋白的含量约占菌体总蛋白的30%,且具有良好的反应原性.GP5蛋白的表达为建立相应的血清学诊断方法奠定了基础.     

猪圆环病毒2型对体外培养仔猪淋巴细胞转化能力及其活性的影响

采集5头PCV2和PRRSV血清抗体阴性的断奶仔猪血液,分离外周血淋巴细胞,分对照组、PHA组、LPS组、PCV2组、PCV2+PHA组、PCV2+LPS组进行体外培养,用MTT法检测PCV2作用2d后对仔猪外周血淋巴细胞转化能力的影响;无菌取脾制成单细胞悬液,分单独淋巴细胞(L)、淋巴细胞+PCV2(L+V)、淋巴细胞与巨噬细胞(L+M)和淋巴细胞与巨噬细胞+PCV2(L+M+V)4组进行体外培养.分别在培养后第0、6、12和24 h收获淋巴细胞,用RT-PCR法检测淋巴细胞IL-2R(α)mRNA的表达.结果显示,与对照组、PHA组和LPS组相比,PCV2组、PCV2+PHA组和PCV2+LPS组淋巴细胞的转化能力均极显著降低(P＜0.01);PCV2对单独培养的淋巴细胞IL-2R(α)表达的影响不显著(P＞0.05),而巨噬细胞能协同PCV2促进淋巴细胞IL-2R(α)的表达,尤其在攻毒后第12和第24 h极显著增加(P＜0.01).证实PCV2在攻毒后第2 d能抑制体外培养T、B淋巴细胞的转化能力,且巨噬细胞能协同PCV2促进T淋巴细胞的活化.     

EXCEL在交通事故分析中的应用

交通安全问题已引起了世界各国的重视,每个国家都在寻求和探索降低交通事故发生率的良策。我国是一个发展中国家,在相当长的一段时间内还会面临着严重的交通安全问题。EXCEL是一种简便、易操作的计算机软件,可以利用EXCEL的数据分析功能,求出道路交通事故中某些推测数据与多因素之间的比例关系,对道路交通事故分析有所帮助。     

Near‐fatal Intoxication with the “New” Synthetic Opioid U‐47700: The First Reported Case in the Czech Republic

Recreational use of the potent synthetic opioid 3,4‐ dichloro‐N‐(2‐(dimethylamino)cyclohexyl)‐N‐methylbenzamide (U‐47700) is rising, accompanied by increasingly frequent cases of serious intoxication. This article reports a case of near‐fatal U‐47700 intoxication. A man was found unconscious (with drug powder residues). After 40 h in hospital (including 12 h of supported ventilation), he recovered and was discharged. Liquid chromatography/high‐resolution mass spectrometry (LC/HRMS) or gas chromatography/mass spectrometry (GC/MS) were used to detect and quantify substances in powders, serum and urine. Powders contained U‐47700 and two synthetic cannabinoids. Serum and urine were positive for U‐47700 (351.0 ng/mL), citalopram (<LOQ), tetrahydrocannabinol (THC: 3.3 ng/mL), midazolam (<LOQ) and a novel benzodiazepine, clonazolam (6.8 ng/mL) and their metabolites but negative for synthetic cannabinoids. If potent synthetic opioids become cheaper and more easily obtainable than their classical counterparts (e.g., heroin), they will inevitably replace them and users may be exposed to elevated risks of addiction and overdose.     

The security of access to accounts under the PSD2

The revised Payment Services Directive (‘PSD2’) has been adopted to stimulate the development of an integrated internal market for payment services. In particular, it facilitates payment initiation services and account information services by granting the providers of these services access to the accounts of the payment service users. At the same time, the recitals state that the PSD2 guarantees a high level of consumer protection, security of payment transactions and protection against fraud.This paper answers the following question: To what extent does the access to accounts of the payment initiation service providers and account information service providers balance the development of the market for payment services with the security of the payment account and the privacy of the user? An analysis of the PSD2 shows that the development of the market for payment services has a higher priority. Security and privacy are ultimately subordinate.First, the PSD2 does not adequately protect the personal data of the users. The definition of ‘account information service’ is broad and covers a wide range of services. This allows the payment service providers to circumvent the limitations of the access to accounts.Next, the payment service providers have a ‘fall back option’ that allows ‘screen scraping’ if the dedicated interface is not functioning properly. Although this access is constrained by several safeguards, the fall back option gives the payment services provider unlimited access to the account of the user.Finally, the payment service providers have considerable freedom to arrange their authentication process as they see fit. The banks seem to be required to trust this process. The PSD2 and regulatory technical standards do not demand that a bank is able to verify the authentication or the integrity of the payment order.     

大鼠局灶性脑挫伤后Cdk5的表达变化

目的研究大鼠受到一定钝力打击造成中度脑损伤后,3h至10d脑组织Cdk5的表达规律及法医学意义。方法建立大鼠钝力性局灶性脑挫伤模型,应用免疫组织化学技术和免疫蛋白印迹技术观察脑损伤后不同时间脑组织中Cdk5表达变化。结果正常组、假手术组脑组织有少量Cdk5表达;实验组损伤后3h组、6h组Cdk5表达增高,12h组表达明显升高(P0.05);在24h达到峰值,3d组、5d组、7d组、10d组逐渐下降,并在7d组、10d组基本降至正常,与正常组、假手术组比较基本无差异性(P0.05)。结论大鼠钝力性脑挫伤后Cdk5在挫伤脑组织周边区表达量增多,并呈现单峰变化,随时间延长基本降至正常。Cdk5表达变化可为早期脑挫伤时间推断奠定基础,提供新的参考指标。     

TGF-β1在SCN5A基因变异所致心源性猝死中调控作用

SCN5A基因变异所致心源性猝死发生机制尚不清楚。目前研究显示TGF-β1介导心肌纤维化以及离子通道重构调节机制的异常可能是SCN5A基因变异导致SUNDS发生的主要原因。本文就转化生长因子β1对SCN5A基因变异所致心源性猝死调控的影响机制研究进展进行综述,以期为心源性猝死法医学研究和实践提供参考。     

"透痹转气法"对佐剂性关节炎大鼠的疗效及滑膜Fas、FasL和Bcl-2蛋白表达的影响

目的:观察"透痹转气法"对佐剂性关节炎(RA)大鼠的影响.方法:复制大鼠RA模型,设立模型组、透痹转气法组(TBZQF)、雷公藤(TPT)组、透痹逐邪胶囊(TBZXC)组;测定各组大鼠足跖肿胀抑制率、关节炎症指数;观察病变关节滑膜病理改变,并检测炎症关节滑膜组织中的Fas、FasL、Bcl-2蛋白表达.结果:与模型组比较,TBZQF组、TBZXC组、TPT组大鼠足跖肿胀抑制率、关节炎指数均显著降低(P＜0.05),而TBZQF组疗效比TBZXC、TPT组更显著(P＜0.05),且起效时间快,疗程短;各治疗组均能改善RA大鼠滑膜充血、水肿,减轻大鼠滑膜炎细胞浸润,而TBZQF组更为明显(P＜0.05);与模型组比较,各治疗组Fas、FasL的表达显著增加,Bcl-2的表达显著减少(P＜0.05),TBZQF与MTX组、TPT组比较,上述指标变化不显著(P＞0.05).结论:TBZQF治疗方案在改善佐剂性关节炎模型大鼠的关节炎症状、抑制炎症方面优于TPT和TBZXC的单一中药治疗,其作用机制可能是通过下调bcl-2基因表达调控Fas/FasL系统,促进增殖的滑膜细胞凋亡.     

猪圆环病毒2型Cap蛋白单克隆抗体的制备与鉴定

利用重组杆状病毒表达的猪圆环病毒2型(PCV2)Cap蛋白免疫BALB/c小鼠,通过杂交瘤技术,共获得6株能稳定分泌抗Cap蛋白抗体的杂交瘤细胞株,分别命名为3H9/1H5、3H9/3C7、4A9/3E2、4A9/3D7、6G7/7B6和6G7/8D3,并通过ELISA、Western-blotting和间接免疫荧光试验(IFA)对6株单抗的特性和抗原表位进行分析鉴定。结果显示,6株杂交瘤细胞培养上清的抗体效价为1∶1 024～1∶2 048,腹水效价为1∶163 840～1∶204 800。3H9/1H5、3H9/3C7、6G7/7B6和6G7/8D3的亚型为IgG1型,4A9/3E2和4A9/3D7为IgG2b型,所有单抗的轻链均为κ型。Western-blotting和IFA结果表明6株单抗均能与重组Cap蛋白和PCV2产生特异性反应。相加ELISA结果显示,6株单抗可能识别不同的抗原表位。为准确鉴别不同单抗针对的抗原位点,以截短表达的重组Cap蛋白为抗原,进行Western-blot鉴定。结果显示,4A9/3E2与4A9/3D7的抗原表位位于101～150aa,其余4株单抗可能针对1～50aa或者天然...