Leukotriene B4 12-hydroxydehydrogenase/15-ketoprostaglandinΔ13-reductase (LTB4 12-HD/PGR) responsible for the reduction of a double-bond of the α,β-unsaturated ketone of an aryl propionic acid non-steroidal anti-inflammatory agent CS-670

CS-670 is a non-steroidal anti-inflammatory agent with an α,β-unsaturated ketone structure. It exerts its pharmacological activity after being transformed to the active metabolite (2S,1′R,2′S)-trans-alcohol. Two consecutive reductions are needed for the formation of the active metabolite, reduction of the double-bond of the α,β-unsaturated ketone moiety, followed by reduction of the resulting saturated ketone. The objective of the current study was to identify the enzyme responsible for reduction of the double-bond. An enzyme purified from rat liver cytosol as a single band on sodium dodecylsulphate-polyacrylamide gel electrophoresis (SDS-PAGE) was analysed by a Mascot database search of nano-LC tandem mass spectrometry (MS/MS) data and the enzyme was identified as 2-alkenal reductase (EC 1.3.1.74), which is known as an β-nicotinamide adenine dinucleotide phosphate (NADPH)-dependent alkenal/one oxidoreductase and has a role for leukotriene B₄ 12-hydroxydehydrogenase/15-ketoprostaglandinΔ13-reductase (LTB₄ 12-HD/PGR). The identification was confirmed by cloning LTB₄ 12-HD/PGR cDNA from rat liver, expressing it in Escherichia coli, and characterizing the properties of the enzyme. The identity was further supported by the subcellular localization in cytosol, a cofactor requirement for NADPH, substrate specificity, and substantial inhibition by 15-ketoPGF_2α, benzylideneacetophenone, indomethacin, and quercitrin. In addition to catalysing the biological reduction of eicosanoids, including prostaglandins, leukotrienes, and lipoxins, LTB₄ 12-HD/PGR was also determined to function as a xenobiotic-metabolizing enzyme.     

Multiple in silico tools predict phenotypic manifestations in congenital thrombotic thrombocytopenic purpura

Congenital thrombotic thrombocytopenic purpura (cTTP) is a rare, recessively inherited genetic disorder with varying clinical presentation that is caused by ADAMTS13 mutations. Several studies have found limited associations between ADAMTS13 mutations and cTTP phenotype. The use of in silico tools that examine multiple mutation characteristics may better predict phenotype. We analysed 118 ADAMTS13 mutations found in 144 cTTP patients reported in the literature and examined associations of several mutation characteristics, including N‐terminal proximity, the evolutionary conservation of the affected amino acid position, as well as amino acid charge/phosphorylation and genetic codon usage to disease phenotype. Structure‐altering mutations were examined for their impact on ADAMTS13 function based on existing ADAMTS13 crystallographic data (AA 77‐685). Our in silico data indicate that: (i) The position of the mutation in the N‐ or C‐terminus, (ii) evolutionary conservation and (iii) codon usage of the affected mutation position are associated with disease parameters, such as age of onset, organ damage and fresh frozen plasma prophylaxis. In conclusion, the usage of multiple in silico tools presents a promising strategy in refining predictions for the diverse presentation of cTTP. Enhancing our utilization of in silico tools to find genotype‐phenotype associations will create better‐tailored approaches for individual patient treatment.     

Normalization of low-density lipoprotein levels and disappearance of xanthomas during pregnancy in a woman with heterozygous familial hypercholesterolemia

Serum lipids and lipoproteins were studied prior to conception, during pregnancy, and after delivery in a woman heterozygous for familial hypercholesterolemia. Prior to conception, serum and low-density lipoprotein (LDL) cholesterol levels were 613 and 528 mg/dL, respectively. At 37-week gestation, serum and LDL cholesterols decreased to the normal levels, 226 and 90 mg/dL, respectively. At two-week postpartum serum and LDL cholesterols returned to the preconception levels, 547 and 427 mg/dL, respectively. At delivery her cutaneous xanthomas almost disappeared. The patient was challenged by ethinyl estradiol of 120 micrograms/d for two months, as a result serum cholesterol decreased from 565 to 385 mg/dL, and LDL cholesterol fell from 460 to 208 mg/dL. During her second pregnancy, serum and LDL cholesterol decreased again significantly. Thus, this case, which showed dramatic reductions of serum and LDL cholesterol levels, may be considered a new variant of heterozygous familial hypercholesterolemia, and the reductions were probably brought about by the action of estrogens, which are known to increase LDL degradation through LDL receptors.     

TACE并IL—2／LAK肝动脉灌注治疗晚期肝癌的初步报告

本文采用ＴＡＣＥ并ＩＬ－２／ＬＡＫ肝动脉灌注治疗晚期肝癌１０例。结果为ＰＲ２例，ＭＲ４例，ＳＤ２例和ＰＤ２例。１０例病人生存期３－１０个月，平均７．８个月。ＩＬ－２／ＡＬＫ灌注后６例ＡＦＰ值明显下降；Ｔ细胞亚群中Ｔｈ数量明显增多，Ｔｓ数量明显显减少，与灌注前比较有显著性差异。ＩＬ２／ＬＡＫ肝动脉灌注未见严重副反应。结果提示ＴＡＣＥ并ＩＬ－２／ＬＡＫ肝动脉灌注为晚期肝癌较好的一种综合治疗方法。     

N-Acetylcysteine for Preventing Pump-Induced Oxidoinflammatory Response During Cardiopulmonary Bypass

Purpose To investigate the effect of N-acetylcysteine on preventing pump-induced oxidoinflammatory response during cardiopulmonary bypass (CPB).Methods Forty patients undergoing coronary artery bypass grafting (CABG) were randomly divided into a study group (n = 20), given 50mgkg^–1 N-acetylcysteine intravenously for 3 days, and a control group (n = 20) given saline. Serum samples were collected for measurement of myeloperoxidase (MPO), malondialdehyde (MDA), interleukin-6, ₁-acid glycoprotein (AAGP), and C-reactive protein (CRP) during surgery and postoperatively.Results The MPO and MDA values showed a similar pattern during and after CPB in the study group, with significantly less variance than in the control group. Interleukin-6 showed similar patterns in the two groups, but the data from 30min after the start of CPB and from 6h post-CPB were significantly different. The AAGP and CRP values were both elevated during CPB in the two groups without a significant difference, but 6 and 24h post-CPB, the values were significantly higher in the control group than in the study group.Conclusions N-Acetylcysteine decreased pump-induced oxidoinflammatory response during CPB, suggesting that it could be a novel therapy for assisting in the prevention of CBP-induced oxidoinflammatory damage.     

TNF-α、LPS、IL-6、PAF与重症胸腹损伤凝血功能障碍的相关性研究

目的 探讨肿瘤坏死因子-α(TNF-α)、内毒素(LPS)、白细胞介素-6(IL-6)和血小板活化因子(PAF)与重症胸腹创伤后凝血功能障碍的相关性与机制.方法 收集2009年1月-2012年6月在解放军第二五三医院急诊科就诊,创伤指数(TI)≥17分,排除合并颅脑损伤及在急诊死亡的胸腹创伤患者82例,在救治同时抽血检查血小板计数(PLT)、部分活化凝血酶原时间(APPT)、凝血酶原时间(PT)、TNF-α、LPS、IL-6、PAF,对检验结果行相关性分析.结果 凝血功能检验结果:PLT:(83.44±38.52)×109/L),APTT:(68.24 ±24.12)S,PT:(28.42±10.83)S;损伤因子检测结果:TNF-α:(36.41±18.09) ng/mL,LPS:(343.66±106.02) IU/L,IL-6:(393.83±143.86) ng/mL,PAF:(15765.31±4431.65) ng/L.PLT与TNF-α、LPS、IL-6、PAF之间相关系数(r)均小于-0.8811,呈显著负相关.APTT、PT与TNF-α、LPS、IL-6、PAF之间r均大于0.9142,呈显著正相关.结论 TNF-α、LPS、IL-6、PAF可能参与了重症胸腹创伤凝血功能障碍的发生过程,对TNF-α、LPS、IL-6、PAF早期干预,或可改善胸腹创伤患者的凝血功能障碍.     

汉族人群白介素17受体C基因单核苷酸多态性与青少年特发性脊柱侧凸的相关性

【摘要】　目的：探讨白介素17受体C（IL-17RC）基因单核苷酸多态性与中国汉族人群青少年特发性脊柱侧凸（adolescent idiopathic scoliosis,AIS）易感性之间的相关性。方法：收集529例AIS女性患者及512例正常同龄女性青少年的静脉血标本,采用聚合酶链反应－限制性片段长度多态性（PCR-RFLP）方法鉴定和统计两组人群IL-17RC基因rs708567和rs279545多态性位点的基因型及等位基因分布频率;比较两组间不同多态性位点各基因型及等位基因分布频率的差异。结果：研究Power值（81%）大于80%,AIS患者组及正常对照组各多态性位点的基因型分布均符合Hardy-Weinberg遗传平衡定律。AIS组rs708567多态性位点GG基因型和G等位基因的分布频率显著高于对照组GG基因型（90.17% vs. 85.55%,P=0.023）和G等位基因（95.1% vs. 92.8%,P=0.028）的分布频率;携带GG基因型青少年中AIS的发病率约为携带AG基因型青少年的1.5倍（OR值=1.55;95% CI:1.45~3.11）。rs279545多态性位点各基因型及等位基因的分布频率在两组间均无统计学差异。结论：中国汉族人群中IL-17RC基因单核苷酸多态性与AIS的发生相关。     

再生小室内间断给予蛋白激酶C激动剂对周围神经表达NGF及损伤修复的影响

目的探讨大鼠坐骨神经再生小室内间断给予蛋白激酶C激动剂-佛波醇酯(phorbol-12-myristate-13-acetate,PMA)后,坐骨神经蛋白激酶C(protein kinase C, PKC)mRNA、神经生长因子(nerve growth factor, NGF)mRNA表达变化规律及轴突数目变化. 方法 SD大鼠42只行双侧坐骨神经中段切断约5 mm,"T"形硅胶管套接,随机分为6组,分别为损伤1、3天,1、2、3和4周组.右侧间断给予1×10-9mol/L PMA(PMA侧);左侧注射等量生理盐水(对照侧).采用组织切片核酸原位杂交(in situ hybridization,ISH)技术检测各组术后各不同点PKC mRNA和NGF mRNA在坐骨神经表达的动态变化及轴突数目变化. 结果对照侧大鼠坐骨神经PKC mRNA在损伤后表达上调,第2周达高峰后下降;NGF mRNA在损伤后表达上调,第3周达高峰值后下降.PMA侧PKC mRNA于第2、3和4周表达较对照侧明显增高,差异有统计学意义(P＜0.01或P＜0.05);NGF mRNA第2、3和4周表达也较对照侧明显增强,差异有统计学意义(P＜0.01).轴突数目在2、3和4周时多,差异有统计学意义(P＜0.01). 结论 PKC介导了周围神经损伤修复中的NGF mRNA表达及神经再生,而且PMA能够促进NGF mRNA表达及促进轴突生长.     

白细胞介素-6对椎间盘细胞中蛋白多糖代谢影响的实验研究

目的:研究白细胞介素-6(interleukin-6,IL-6)对椎间盘纤维环和髓核细胞中蛋白多糖代谢的影响。方法:自然流产的胎儿4例,4h内无菌取出椎间盘,分别进行纤维环和髓核细胞的体外培养。在纤维环细胞的培养液中分别加入IL-60(对照组)、400、800ng/ml,培养24h后,用Alcian法检测培养液中硫酸软骨素的含量。在培养中的髓核细胞中加入IL-60(对照组)、100、400、800ng/ml,培养24h,然后测量培养液中硫酸软骨素的含量。结果:在纤维环细胞中加入IL-6组较不加IL-6对照组培养液中硫酸软骨素的含量增加,在髓核细胞组中加入IL-6组和对照组的差别无显著性。结论:IL-6可以刺激椎间盘纤维环细胞中蛋白多糖的合成,但对髓核细胞中蛋白多糖的合成没有明显的作用。