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.     

"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。     

Webcasters’ protection under copyright – A comparative study

This paper studies the fourth generation ‘peer-2-peer’ (P2P) file sharing technology which is also known as streaming technology from the technology point of view. It critically analyses the nature of streaming technology and assesses similarities to broadcasting for the purpose of copyright protection in the name of webcasting. In addition, it considers the possibility of protecting webcasting within the framework of broadcasting under copyright and the extent of broadcasters’ rights applicable to webcasters. Based on the conclusions derived from the discussion of UK, New Zealand, Italian and US law on streaming, WIPO Standing Committee on Copyright and Related Rights (SCCR) draft documents will be critically analyzed. The Last part of this work will offer suggestions as to how to protect webcasting (streaming technology) under the umbrella of copyright.     

大鼠闭合性脑损伤后MAP-2的时序性表达

目的探讨大鼠闭合性颅脑损伤后脑组织微管相关蛋门－2（MAP－2）表达变化规律。方法建立大鼠闭合性颅脯挫伤模型,64只Wistar大鼠随机分为损伤后0．5h、6h、12h、ld、3d、7d、14d7个损伤组及对照组,每组8只。应用免疫组化和蛋h印记Western blot法及图像分析技术,检测脑挫伤后各设定时间点挫伤脯组织中MAP－2的变化。结果免疫纽化染色显示：对照组大鼠脑组织中可见MAP－2阳性表达;实验组伤后0．5h,挫伤灶及周同MAP－2阳性表达下降,伤后6h阳性表达降至最低,伤后12h,阳性表达逐渐增多,伤后14d,仍保持较高水平;蛋白印记结果与免疫组化结果一致。统计学分析显爪：MAP－2的表达各损伤组与对照组比较,差异均有统计学意义（P〈0．05）,伤后12h、3d及14d与上…组比较,差异有统计学意义（P〈0．05）。结论大鼠脑挫伤后14d内,MAP－2存脑组织挫伤灶及其周同呈现先减少后逐渐增多的表达变化,可作为推断脑挫伤经过时间的参考指标之一。