Evaluation of the monocyte activation test for the safety testing of meningococcal B vaccine Bexsero: A collaborative study

The aim of this collaborative study was to evaluate the robustness of the monocyte activation test (MAT) for quantifying the pyrogenic content in the outer membrane vesicle (OMV)-containing vaccine Bexsero: the first meningococcal B vaccine to be licenced. We analysed datasets from 9 laboratories covering 15 test systems for 3 batches of Bexsero with higher, equivalent and lower activity relative to a reference lot in the MAT. Activity was measured in terms of relative pyrogen units (RPU) based on European Pharmacopoeia (Ph. Eur.) MAT Chapter 2.6.30 Method C: Reference Lot Comparison Test. We report that all 15 test systems were consistent in that they showed sample A to be the most active in the MAT; that 13 of 15 test systems had an accuracy of more than 80% and an overall geometric mean RPU of 1.03 with lower and upper 95% confidence limits of 0.97 and 1.09 respectively for a sample with an expected value of 1.00 RPU. We also report larger variability in the results for test systems involving cells from individual blood donations for sample A suggesting that there could be donor to donor differences in sensitivity to the vaccine constituents responsible for the higher activity of this batch. Overall, the consistency and accuracy of the MAT was remarkable given the range of test systems used by participants, all of which are permitted by the Ph. Eur. General MAT Chapter. This is important given the limitations of the rabbit pyrogen test for the control of pyrogenicity in general and particularly with products with intrinsic pyrogenicity such as Bexsero.     

发热及小儿解热药的使用

发热是对抗疾病的一种复杂生理学反应,涉及细胞因子介导的体温升高,急性期反应物的生成,内分泌和免疫学系统的激活。内生性致热原和众多体温中枢调节介质构成了一个复杂的体温调节网络,负责机体的体温调节平衡。发热是小儿临床中最常见的症状之一,目前临床中使用的小儿解热药大多具有副作用,所以了解发热的基本机制,开发安全有效的小儿解热药具有重要的临床意义。     

内源性分泌型糖基化终产物受体与2型糖尿病颈动脉粥样硬化的关系

目的 探讨内源性分泌型糖基化终产物受体(esRAGE)水平与2型糖尿病(T2DM)颈动脉粥样硬化的关系.方法 60例T2DM患者,分成颈动脉粥样硬化组(A组,30例)和无颈动脉粥样硬化组(B组,30例).28例非糖尿病者为对照组(C组).分别检测esRAGE浓度、体重指数、血压、血脂和糖化血红蛋白(HbA1c).结果 A组和B组esRAGE浓度均明显低于C组(P＜0.05),A组明显低于B组(P＜0.01).多危险因素与T2DM颈动脉粥样硬化Logistic回归分析表明,esRAGE是T2DM颈动脉粥样硬化的重要保护因素(b＜0,OR＜1,P＜0.1).结论 esRAGE与T2DM密切相关,对T2DM颈动脉粥样硬化有潜在保护作用.     

超滤和活性炭在注射剂生产工艺中的应用比较

目的比较超滤工艺和活性炭工艺在普通注射剂生产中去除热原的优缺点以及对产品质量的影响。方法①配制浓度为1000EU.mL-1的内毒素指示剂水溶液,分别考察超滤工艺和活性炭工艺去除内毒素指示剂的效果;②分别考察超滤工艺和活性炭工艺在10%的维生素C药液、3%的曲克芦丁药液、0.04%的法莫替丁药液中去除热原的效果及对产品质量的影响。结果①截留分子量为10000的超滤能有效的去除药液中的热原、微粒、病毒等异物,对产品的色泽和澄清度均有一定的改善作用,但对不同产品标示量的均一性会产生影响;②活性炭对产品的色泽有一定的改善,但去除热原的效果不明显,对药品主成分会产生一定的吸附作用。结论超滤去除热原的作用[1]和对产品质量的影响均要优于活性炭。     

Vinyl Chloride—A Classical Industrial Toxicant of New Interest

The carcinogenicity of vinyl chloride in humans was recognized in 1974 based on observations of hepatic angiosarcomas in highly exposed workers. A multiplicity of endpoints has been demonstrated. The primary target organ, the liver, displays differential susceptibilities of hepatocytes and sinusoidal cells, which are modified by factors of age and dose. There is consistency in organotropism between experimental animals and humans. Vinyl chloride is a pluripotent carcinogen, predominantly directed toward hepatic endothelial (sinusoidal) cells, and second toward the parenchymal cells of the liver. The similarity of results between experimental animals and humans is a solid basis of an amalgamation of experimental and epidemiological risk estimates. Vinyl chloride requires metabolic activation for carcinogenicity and mutagenicity, and toxicokinetics are a key to interpret the dose response. Practically the entire initial metabolism of vinyl chloride is oxidative. At higher exposure concentrations this is nonlinear, and metabolic saturation of metabolism in rats is reached at about 250 ppm. This is consistent with the plateau of hepatic angiosarcoma incidence in rat bioassays. Physiologically based pharmacokinetic/toxicokinetic (PBPK) models have been developed and successfully applied within the frame of human cancer risk assessments. The major DNA adduct induced by vinyl chloride (～ 98% of total adducts in rats), 7-(2-oxoethyl)guanine, is almost devoid of promutagenic activity. The clearly promutagenic “etheno” adducts N²,3-ethenoguanine and 3,N⁴-ethenocytosine each represent ～ 1% of the vinyl chloride DNA adducts in rats, and 1,N⁶-ethenoadenine is found at even lower concentrations. Etheno adducts appear to have a long persistence and are repaired by glycosylases. Vinyl chloride represents a human carcinogen for which a series of mechanistic events connects exposure with the carcinogenic outcome. These include (1) metabolic activation (to form chloroethylene oxide), (2) DNA binding of the reactive metabolite (to exocyclic etheno adducts), (3) promutagenicity of these adducts, and (4) effects of such mutations on protooncogenes/tumor suppressor genes at the gene and gene product levels. In rat hepatocytes, a further event is a biomarker response. Cancer prestages (enzyme-altered foci), as quantitative biomarkers, provide a tool to study dose response even within low dose ranges where a carcinogenic risk cannot be seen in cancer bioassays directly. Such biomarker responses support a linear nonthreshold extrapolation for low-dose assessment of carcinogenic risks due to vinyl chloride. Published risk estimates based on different sets of data (animal experiments, epidemiological studies) appear basically consistent, and on this basis an angiosarcoma risk of ～ 3 × 10^?4 has been deduced by extrapolation, for exposure to 1 ppm vinyl chloride over an entire human working lifetime. An important point that should be considered in regulatory standard settings is the presence of a physiological background of those etheno DNA adducts, which are also produced by vinyl chloride. Likely reasons for this background are oxidative stress and lipid peroxidation. In essence, fundamentals of the hepatocarcinogenicity of vinyl chloride appear now well established, providing a solid scientific basis for regulatory activities.     

内源性药物碘制剂的生物等效性研究

目的：以卵磷脂络合碘的人体生物等效性研究为例,探讨内源性药物的生物等效性研究方法。方法：对内源性及其他来源（饮食性）的碘进行控制。24名健康受试者进行双周期、双交叉、单剂量口服9片卵磷脂络合碘试验品和参比品,采用砷铈催化分光光度法测定血、尿中碘的浓度,用DAS2.1.1软件计算药动学参数,并进行生物等效性检验。结果：试验品与参比品的血碘平均净浓度增加值分别为16.8%和19.0%,尿碘回收率分别为84.5%和83.3%。计算同一周期的尿碘净排泄量,用尿碘累积净排泄量（Ae0-t）、最大净排泄速率（Rmax）进行生物等效性分析。本实验研究表明卵磷脂络合碘受试制剂与参比制剂具有生物等效性。结论：具有生物自稳定机制的内源性药物的生物等效性研究,实验设计更加复杂,各种内外因素对结果影响较大,需要尿药浓度结合背景扣除。     

罗布麻叶对大鼠尿液内源性物质代谢的影响

目的：采用代谢组学方法考察罗布麻叶对正常大鼠尿液内源性物质代谢的影响。方法：罗布麻叶用10倍量70%乙醇回流提取,将大鼠分为空白对照组、罗布麻叶高剂量组及低剂量3组,连续灌胃给药29 d,运用SIMCA-P软件中的PCA法与PLS-DA法对比分析。结果：罗布麻叶给药组与空白对照组得分点达到很好的区分效果,连续给药22 d时,尿液内源性物质代谢改变最为明显。相比于罗布麻叶低剂量组,高剂量组与空白对照组之间差距更大。结论：罗布麻叶提取物对正常大鼠机体内源性物质代谢产生了影响,为进一步阐释罗布麻叶的药性研究工作提供了依据。     

茴香注射液热原检查方法学研究

目的:验证红茴香注射液热原检查的适用性。方法:参考《中国药典》2010年版一部附录中药注射剂安全性检查法应用指导原则,对红茴香注射液进行热原检查的方法学验证。结果:红茴香注射液原液在剂量为1 mL.kg-1有一定的毒性反应,不适宜进行热原试验;10倍稀释液对热原检查无干扰。结论:可用热原检查法检查红茴香注射液中可能存在的热原,浓度为其10倍稀释液,剂量为1 mL.kg-1。     

Pharmacological characteristics of agonistantagonist analgesics

A brief history and pharmacological characteristics of agonist-antagonist analgesics are presented. The importance of this class of compounds on the development of opioid-receptor concepts is described.     

肝脏缺血再灌注损伤的ceRNA网络构建和潜在治疗药物筛选

背景与目的 肝脏缺血-再灌注（I/R）损伤是肝移植和肝切除手术过程中常涉及的共同病理生理变化。竞争性内源性RNA（ceRNA）调控网络可参与多种疾病的发生发展。然而ceRNA网络在肝脏I/R损伤中的功能仅有少量报道。本研究旨在应用生物信息学方法构建与肝脏I/R损伤相关的ceRNA网络,同时基于差异表达基因筛选潜在治疗药物。方法 从GEO数据库获取肝脏I/R损伤的mRNA及miRNA表达芯片数据。使用R语言中的limma包进行基因差异表达分析,并使用ggplot2包进行散点图、火山图和热图绘制。使用String数据库及Cytoscape软件进行蛋白互作（PPI）网络构建。利用Metascape数据库对筛选出的差异mRNA进行GO/KEGG功能富集分析。通过转录调控网络数据库分析可能调控这些差异基因的转录因子。使用miRTarBase数据库构建miRNA-差异表达基因网络。通过starBase：ceRNA数据库构建ceRNA网络。使用比较毒物基因组学数据库（CTD）筛选对关键差异基因表达具有潜在作用的天然药物。结果 从GEO数据库获得2个肝脏I/R损伤mRNA数据集（GSE10654和GSE117066）和1个肝脏I/R损伤miRNA数据集（GSE72315）。通过limma包及Venn图分析mRNA表达数据集,筛选到16个在I/R组上调表达,在缺血后适应（IPO）组下调表达的基因;7个在I/R组下调表达,在IPO组上调表达的基因。GO/KEGG功能富集分析结果显示差异基因主要参细胞死亡的正调控及对细胞外刺激反应的生物学过程,并参与MAPK信号通路。转录调控网络数据库分析获得6个转录因子（Trp53、Cebpb、Crebbp、Fos、Nfkb1及SP1）可能参与这些差异基因的调控。通过miRTarBase数据库分析,并结合GSE72315数据集中miRNA在I/R损伤后的差异表达,获得两个可能在肝脏I/R损伤中发挥重要的作用miRNA-mRNA轴（mmu-miR-32-5p-Btg2与mmu-miR-9-5p-Mt2）。通过starBase：ceRNA数据库分析,最终获得9条ceRNA网络,分别是：XIST/MEG8/LINC00963/MALAT1-miR-32-5p-Btg2轴、XIST/NEAT1-miR-132-3p-Btg2轴及HSPA9P1/RALGAPA1P1/RPS26P39-miR-9-5p-Dusp6轴。CTD数据库筛选到7种植物药（槲皮素、白藜芦醇、染料木黄酮、香豆雌酚、姜黄素、辣椒素及东莨菪碱）可降低关键基因的表达发挥潜在治疗作用。结论 通过生物信息学方法筛选了肝脏I/R损伤过程中的关键ceRNA网络及治疗的潜在天然药物,为进一步研究肝脏I/R损伤的发病机制及治疗药物提供重要依据。