用于治疗囊性纤维化的药物Lumacaftor

囊性纤维化(CF)是一种严重威胁生命的遗传性疾病,该病系由CF跨膜传导调节蛋白(CFTR)的缺陷所引起。正常的CFTR为1种存在于细胞膜上的氯离子通道蛋白,氯离子可通过此通道蛋白自由进出细胞,细胞也可分泌1层稀薄的黏液以保护气道上皮。若cftr基因出现G551D突变,CFTR的通道就不能正常开放,氯离子也就无法自由地进出细     

治疗分泌型腹泻的药物初露端倪

美国加州大学Verkman领导的国际研究小组最近确认了一种称为纤维囊性跨膜转导调节因子(CFTR)的物质 ,它可以阻止与分泌型腹泻和纤维囊性病相关的氯离子跨膜转运蛋白的功能。正常的CFTR蛋白允许氯离子分泌到肠腔或呼吸道粘性分泌物中 ,所产生的渗透压可使水分从血液中渗出。在纤维囊性病的病人中 ,CFTR的突变能阻止氯离子的有效运输。在分泌型腹泻 ,感染性病原体诱发的氯离子分泌增加能造成病人脱水。这是发展中国家里 ,造成 5岁以下儿童死亡的主要原因。　　Verkman等使用细胞荧光分析技术筛选了5 0 0 0 0个化合物 ,从中选择了最能抑…     

双载体断裂CFTR基因转移及翻译后的连接和功能

囊性纤维化跨膜电导调节体 (CFTR) 基因突变导致一种常染色体隐性遗传病囊性纤维化 (CF), 利用腺辅助病毒 (AAV) 载体转运CFTR基因的基因疗法受到AAV载体容量的限制。本文采用intein的蛋白质反式剪接技术, 以双载体转运CFTR基因, 研究了于其调节结构域 (R) 断裂成两部分的CFTR基因翻译后的连接及其产生的Cl⁻通道功能。将人CFTR cDNA于R结构域的Ser⁷¹²密码子前断裂, 构建一对融合Ssp DnaB intein编码序列的真核表达载体。将这对载体共转染培养的幼年仓鼠肾 (BHK) 细胞, 通过瞬时表达, 全细胞和单通道膜片钳记录Cl⁻电流, 并用Western blotting观察CFTR蛋白的剪接。结果表明, 共转染细胞显示较高的全细胞Cl⁻电流和单个Cl⁻通道开放活性, 说明CFTR的Cl⁻通道功能的恢复, 用CFTR特异性抗体进行的细胞总蛋白Western blotting显示有完整的CFTR蛋白条带形成, 表明intein可有效连接翻译后的两部分CFTR蛋白。结果提示, 蛋白质剪接技术可有效用于双载体系统转运CFTR基因, 为进一步应用双AAV载体转运CFTR基因的CF基因治疗研究提供了实验依据。     

囊性纤维跨膜转运调节因子蛋白在豚鼠内耳的表达及意义

目的 检测氯离子通道蛋白-囊性纤维跨膜转运调节因子(CFTR)在豚鼠内耳中的表达,探讨其在内耳液体平衡调节中的意义.方法 利用豚鼠内耳水肿模型,采用免疫组化方法检测CFTR蛋白表达情况.结果 CFTR在水肿内耳及正常内耳不同部位表达情况不同:其中在耳蜗血管纹上皮细胞、内淋巴囊、椭圆囊、球囊有强表达,在基底膜鼓阶面细胞、螺旋韧带纤维细胞、螺旋神经节细胞表达较弱,水肿内耳中CFTR的表达均明显高于正常内耳,差异有统计学意义(P＜0.05).结论 CFTR在内耳水、电解质调节方面具有较为重要的作用,其深入研究探讨对内耳水肿疾病发生机制的了解将有重要的临床意义.     

丹皮酚对囊性纤维化跨膜电导调节因子氯离子通道的激活作用

目的:研究丹皮酚对依赖于3’,5’-环腺苷酸(cAMP)的囊性纤维化跨膜电导调节因子(CFTR)Cl-通道的激活作用。方法:利用Cl-通道细胞荧光测定模型进行测定丹皮酚对CFTR介导的I-内流速度。结果:丹皮酚对野生型和ΔF508突变型CFTR Cl-通道均具有激活作用,而对G551D突变型CFTR Cl-通道无激活作用。丹皮酚在发挥激活作用时具有快速、可逆的特点并依赖于腺苷环化酶激动剂Forskolin的存在。初步确定了丹皮酚通过直接与CFTR结合发挥作用。结论:研究提示CFTR可能是某些中药降压作用药靶之一。此外,本实验为传统中药的降压作用的分子药理学机制作了初步探讨。     

牡丹皮对脂多糖诱导大鼠系膜细胞的保护作用

目的 观察牡丹皮对脂多糖（LPS）诱导的大鼠系膜细胞（GMC）转化生长因子-β1（TGF-β1）、血小板衍生性生长因子（PDGF）和血管内皮生长因子(VEGF)表达的影响。方法 用脂多糖 (20 mg·L^-1)刺激大鼠系膜细胞,经牡丹皮(50、100、200 mg·L^-1)培养48 h后,采用ELISA法检测转化生长因子-β1的分泌水平;实时定量PCR方法检测血小板衍生性生长因子和血管内皮生长因子 mRNA的表达。结果 牡丹皮可以抑制脂多糖诱导的大鼠系膜细胞转化生长因子-β1的分泌,下调血小板衍生性生长因子和血管内皮生长因子的表达。结论 牡丹皮可能通过减少转化生长因子-β1的分泌,抑制血小板衍生性生长因子和血管内皮生长因子的表达,对受损的大鼠系膜细胞起保护作用。     

临床药师参与1例间变性大细胞淋巴瘤患儿严重剥脱性皮炎的病例分析

目的 分析一例间变性大细胞淋巴瘤患儿产生严重剥脱性皮炎的原因,探索临床药师在指导用药中的作用。方法 系统分析1例严重剥脱性皮炎患儿的给药、皮炎发生过程、处理及预后。结果 间变性大细胞淋巴瘤及联合用药均可引起剥脱性皮炎。结论 临床药师参与患者的治疗与监护,可减少不良反应的发生,保障患者的用药安全。     

木犀草素促进囊性纤维化跨膜电导调节因子氯离子通道开放

目的：研究木犀草素对依赖于3，5-环腺苷酸（cAMP）的囊性纤维化跨膜电导调节因子（CFTR）Cl^-通道的激活作用。方法：利用稳定共表达人CFTR与一种对卤族元素敏感的荧光绿蛋白突变体（EYFP-H148Q）的Fischer大鼠甲状腺（FRT）细胞模型，测定木犀草素对CFTR介导的I^-内流速度。结果：木犀草素能够以剂量依赖的方式激活CFTR的I^-转运，其对CFTR Cl^-通道的激活效应具有作用迅速、可逆的特点。木犀草素在发挥激活作用时依赖于腺苷环化酶激动剂Forskolin的存在，其单独与细胞孵育不提高细胞内cAMP的水平。结论：发现了木犀草素能够以剂量依赖方式激活CFTR，并且初步确定了木犀草素通过直接与CFTR结合发挥作用。     

柴枳夏及汤对胆汁反流性胃炎大鼠胃黏膜的保护作用及机制研究

目的 探讨柴枳夏及汤对实验性胆汁反流性胃炎（BRG）大鼠胃黏膜的保护作用及机制。方法 将Wistar大鼠按照体质量及性别以随机数字表法分为5组：对照组、模型组、铝碳酸镁片（阳性药）组和柴枳夏及汤低、高剂量（生药量12.77、25.54 g/kg）组,除对照组外,ig自制反流液制备大鼠BRG模型。观察对照组及模型组大鼠进食、大小便、毛色、体质量等;肉眼及HE染色后于光镜下观察胃窦黏膜病理组织学变化;ELISA法检测血清GAS及胃窦黏膜PGE2水平。结果 与对照组比较,模型组大鼠出现排稀糊便、便中含红黄色黏液、反应迟钝、体质量显著下降（P<0.05）等特点;肉眼可见胃窦部点片状糜烂,有黄绿色胆汁及较多黄色黏液;胃窦黏膜出现大量炎性细胞浸润和肠上皮化生,炎症及肠上皮化生积分显著提高（P<0.01）;GAS及PGE2水平明显下降（P<0.01）;与模型组比较,柴枳夏及汤高剂量明显改善胃黏膜病理组织损伤形态;减轻炎性细胞浸润和肠上皮化生,显著降低积分（P<0.05、0.01）;显著提高GAS及PGE2水平（P<0.01）。结论 柴枳夏及汤具有明显的保护胃黏膜的作用,其机制可能与调节血清GAS、胃窦黏膜PGE2水平有关。     

囊性纤维化治疗药依伐卡托的研制

<正>1研发背景囊性纤维化是人类遗传性疾病,由于特定基因的缺失或突变,使体内多种器官功能逐渐消退,例如导致呼吸和行动困难以及提早死亡。囊性纤维化疾病是由于编码囊性纤维化跨膜传导调节因子(cystic fibrosis transmembrane conductance regulator, CFTR)的基因发生突变,蛋白中Phe508残基缺失,使CFTR蛋白折叠、运输和膜稳定性发生变化。CFTR构成了蛋白激酶A(PKA)调控的阴离子通道,调控许多器官上皮细胞的氯离子和碳酸氢根离子的转运,因而CFTR基因变异往往影响全身的机能。囊性纤维化是一类凶恶的罕见病,没有有效的治疗药物。     

Interindividual variability in metabolic activation in humans in vivo

In assessing interindividual variability in metabolic activation, the toxic metabolite is often too unstable for conventional analysis. Possible alternatives include a stable product of the reactive metabolite e.g. cysteinyl derivatives of N-acetyl-4-benzoquinoneimine, the toxic metabolite of paracetamol, adducts with DNA or protein, and indirect measurement of the activity of the enzyme(s) producing the active metabolite. An example of the last approach is the use of furafylline, a highly specific inhibitor of human CYP1A2, to determine the extent of the metabolic activation of the cooked food mutagens PhIP and MeIQx. The extent of inhibition, determined from levels of unchanged amine in urine, is an indirect measure of the activity of the activation pathway. Further refinement of this approach, allied to improved measures of the biological process of interest should prove of value in evaluating interindividual variability and its role in the risk assessment process.     

Quantitative in vivo and in vitro sex differences in artemisinin metabolism in rat

1. The pharmacokinetics of the antimalarial compound artemisinin were compared in the male and female Sprague-Dawley rat after single dose i.v. (20 mg.kg) or i.p. (50 mg.kg) administration of an emulsion formulation. 2. Plasma clearance of artemisinin was 12.0 (95% confidence interval: 10.4, 13.0) l.h. kg in the male rat and 10.6 (95% CI: 7.5, 15.0) l.h. kg in the female rat suggesting high hepatic extraction in combination with erythrocyte uptake or clearance. Artemisinin half-life was 0.5 h after both routes of administration in both sexes. Values for plasma clearance and half-lives did not statistically differ between the sexes. 3. After i.p. administration artemisinin AUCs were 2-fold higher in the female compared with male rat (p 0.001). Artemisinin disappearance was 3.9-fold greater in microsomes from male compared with female livers and it was inhibited in male microsomes by goat or rabbit serum containing antibodies against CYP2C11 and CYP3A2 but not CYP2B1 or CYP2E1. 4. The unbound fraction of artemisinin in plasma was lower (p 0.001) in plasma obtained from the male (8.8 2.0%) compared with the female rat (11.7 2.2%). 5. The possibility of a marked sex difference, dependent on the route of administration, has to be taken into account in the design and interpretation of toxicological studies of artemisinin in this species.     

Theophylline kinetics in peripheral tissues in vivo in humans

Several biochemical and cellular effects have been described for methylxanthines under in vitro conditions. However, it is unknown, whether threshold concentrations required to exert these effects are attained in target tissues in vivo. We therefore employed the microdialysis technique for measuring theophylline concentrations in peripheral tissues under in vivo conditions.Following in vitro and in vivo calibration, microdialysis probes were inserted into the medial vastus muscle and into the periumbilical subcutaneous adipose layer of healthy volunteers. Following single oral dose administration of 300 mg or i.v. infusion of 240 mg theophylline, in vivo time courses of theophylline concentrations were monitored in tissues and plasma. Major pharmacokinetic parameters (c_max, t_max, AUC) were calculated for plasma and tissue time courses. The mean AUC_tissue /AUC_plasma-ratio was 0.56 (p.o.) and 0.55 (i.v.) for muscle and 0.55 (p.o.) and 0.72 (i.v.) for subcutaneous adipose tissue.We conclude that microdialysis provides important information on the distribution and the tissue pharmacokinetics of theophylline.Abbreviations FPIA Fluorescence polarisation immuno assay - AUC Area under the curve - t_max Time to peak concentration - c_max Peak concentration     

休克时血中氧自由基的变化

本实验测定10名休克患者血浆和红细胞的丙二醛(MDA)、血浆总抗的氧化活性(AOA)的含量。结果表明:休克病人红细胞膜和血浆 MDA 含量(4.298±0.722;5.348±0.834)与对照组(3.235±0.682;4.356±1.081)比较明显增高(P<0.05);血浆 AOA(39.65±7.858)与对照组(48.21±10.81)比较明显降低(P<0.01)。提示:休克时,患者机体内自由基反应增强是引起组织细胞损伤的原因之一。     

Polymorphisms in genes involved in neurotransmission in relation to smoking

Smoking behavior is influenced by both genetic and environmental factors. The genetic contribution to smoking behavior is at least as great as its contribution to alcoholism. Much progress has been achieved in genomic research related to cigarette-smoking within recent years. Linkage studies indicate that there are several loci linked to smoking, and candidate genes that are related to neurotransmission have been examined. Possible associated genes include cytochrome P450 subfamily polypeptide 6 (CYP2A6), dopamine D₁, D₂, and D₄ receptors, dopamine transporter, and serotonin transporter genes. There are other important candidate genes but studies evaluating the link with smoking have not been reported. These include genes encoding the dopamine D₃ and D₅ receptors, serotonin receptors, tyrosine hydroxylase, trytophan 2,3-dioxygenase, opioid receptors, and cannabinoid receptors. Since smoking-related factors are extremely complex, studies of diverse populations and of many aspects of smoking behavior including initiation, maintenance, cessation, relapse, and influence of environmental factors are needed to identify smoking-associated genes. We now review genetic polymorphisms reported to be involved in neurotransmission in relation to smoking.     

Tramadol concentrations in blood and in cerebrospinal fluid in a neonate

Based on blood and cerebrospinal fluid samples collected in a full-term neonate, the penetration of tramadol in the central nervous system is described. Following intravenous administration of tramadol, a lag time of about 4 h was observed until full blood–brain equilibration was achieved. This pharmacokinetic observation is in line with a recent pharmacodynamic evaluation of the central opioid effects of tramadol in adults.     

Disease control in asthmatic children seen in private practice in Switzerland

ABSTRACT

Background: Asthma is the most common chronic childhood disease in Switzerland with a prevalence of 10%. Asthma has a high economic burden accounting for high medical costs. Assessment of disease control is likely to be of help in the implementation of strategies to improve asthma. Therefore, we aimed to evaluate asthma control and therapy regimens among children in private practice.

Methods: We assessed asthma control as well as therapy regimens in 575 asthmatic children in an experience programme in Switzerland by using an abbreviated questionnaire based on the asthma control questionnaire and the child health questionnaire on Visit 1 and Visit 2.

Results: Good asthma control at Visit 1 was only present in 25.7% of asthmatic children. Occasional asthma symptoms, limitation of physical activity, nocturnal awakening and anxiety of the parent was present in 80.5%, 41.2%, 46.8% and 57% of the children, respectively. After adjustment of therapy regimens at Visit 1, mainly by adding a leukotriene receptor antagonist, asthma control was reported to be much better in 53.4% of the children at Visit 2.

Conclusions: As asthma control is inadequately achieved within a major portion of asthmatic children, it is imperative to find measures to improve asthma control and hence, to reduce the burden of disease.