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。     

五种重要猪病病毒基因芯片探针的建立及其应用

将猪繁殖与呼吸综合征病毒(PRRSV)、猪瘟病毒(CSFV)、猪细小病毒(PPV)、猪伪狂犬病痛毒(PRV)、猪圆环病毒2型(PCV-2)的特异cDNA片段作为探针点制于氨基修饰的玻片上,以这5种细胞毒的核酸作为模板,进行不对称PCR扩增,制备靶物,经绿色荧光染料Cy3标记后,分别与cDNA微阵列的芯片探针进行杂交,杂交信号经Genepix 4000A扫描仪扫描.结果显示,该芯片探针可以特异地与Cy3标记的这5种靶物结合.用该基因芯片对100份现地采集的猪全血样品进行检测,检出PRRSV阳性样品26份,PCV-2阳性样品47份,PRRSV和PCV-2混合感染阳性样品17份,PPV阳性样品5份,PRV阳性样品2份,CSFV阳性样品20份.表明,本试验研制的cDNA芯片探针可以特异地检测这5种猪病病毒,具有良好的诊断应用前景.     

基于EPO介导的JAK2/STAT5信号通路研究百会、大椎刺络促缺血性脑卒中后血管新生的调控机制

目的 基于促红细胞生成素（erythropoietin,EPO）介导的JAK2/STAT5信号通路探讨百会、大椎刺络促缺血性脑卒中后血管新生的调控机制。方法 选取健康SD雄性大鼠150只,随机分为正常对照组、假手术组、模型对照组、刺络治疗组、刺络+EPO拮抗组,每组30只;正常对照组不进行手术,假手术组仅分离血管而不插线栓,模型对照组、刺络治疗组、刺络+EPO拮抗组采用改良的Zea-Longa法制备大脑中动脉局灶性脑缺血再灌注模型（middle cerebral artery occlusion/reperfusion model,MCAO/R）大鼠。模型制备后刺络治疗组、刺络+EPO拮抗组采用“百会”“大椎”穴刺络进行干预,刺络+EPO拮抗组在针刺前将脂质体-siEPO2复合物按5 mg/kg剂量对大鼠进行腹腔注射,使之进入合成EPO的靶细胞,在细胞内引起针对EPO的RNA干扰效应。采用苏木精-伊红染色观察各组大鼠脑皮质病理形态,RT-PCR法检测各组大鼠脑皮质中EPO、血管内皮生长因子（vascular endothelial growth factor,VEGF） mRNA表达水平,Western Blot法检测各组大鼠脑皮质中P-JAK2、P-STAT5、VEGF蛋白表达水平。结果 与正常对照组比较,模型对照组及刺络+EPO拮抗组大鼠脑皮质病理损伤严重;与模型对照组比较,刺络治疗组大鼠脑皮质病理损伤明显改善。与正常对照组比较,模型对照组大鼠脑皮质中EPO、VEGF mRNA表达水平显著升高（P＜0.05）,P-JAK2、P-STAT5、VEGF蛋白表达水平显著升高（P＜0.05）,刺络治疗会进一步增强这种趋势;与刺络治疗组比较,刺络+EPO拮抗组大鼠脑皮质中EPO、VEGF mRNA表达水平降低（P＜0.05）,P-JAK2、P-STAT5、VEGF蛋白表达水平降低（P＜0.05）。结论 通过百会、大椎刺络能上调MCAO/R模型大鼠EPO的表达水平,激活JAK2/STAT5信号通路,促进下游VEGF的表达而促进缺血性脑卒中后血管的新生。     

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.     

iCLIC Data Mining and Data Sharing workshop: The present and future of data mining and data sharing in the EU

Held at Southampton University's Highfield campus and hosted by iCLIC, an interdisciplinary core on Law, the Internet and Culture, the Data Mining and Data Sharing workshop brought together attendees and speakers from industry, government, academia and a range of disciplines alike. The workshop comprised two sessions, each with a keynote and an associated panel. The first session was chaired by Eleonora Rosati and dealt with copyright and database rights, data mining and data sharing. The second session, chaired by Sophie Stalla-Bourdillon, focussed on data protection, data mining and data sharing. The following report covers both sessions, associated panel discussions and the subsequent question and answer sessions.     

"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...     

适合猪圆环病毒2型敏感复制的PK15细胞株的克隆与鉴定

为了有效提高猪圆环病毒2型(PCV2)在PK15细胞中增殖的效价,采用有限稀释法将PK15细胞进行克隆,通过间接免疫荧光方法筛选对PCV2敏感的克隆细胞.结果表明,通过克隆后获得1株对PCV2高度易感的细胞PK15-B1,该细胞接种PCV2后培养3 d,病毒的TCID50达10-7.0/mL,病毒效价和生产速度明显高于...     

宗教自由基本权利的肥大症——关于宗教和世界观自由与一般权利有效性请求的关系

当人们通览宗教和世界观自由的基本权利在过去十几年里的实际运用和法教义学上的发展时,很难忽略其在规范性轮廓上萎缩这一事实。这一现象由构成要件要素的发展上更多地考虑主观标准所承载,并且经由联邦宪法法院在司法判例中对有关限制问题的决断性的、限制性的判决而被强化。因此,相对于依据宗教或者世界观的动机而提起的个人的发展请求,一般的法治国规范的有效领域持续地受到限制。当相关现象的潜在冲突在德国呈现趋势性增长的时候,这样的判断就显得越发的危险。