Intracellular triggering of Fas aggregation and recruitment of apoptotic molecules into Fas-enriched rafts in selective tumor cell apoptosis

We have discovered a new and specific cell-killing mechanism mediated by the selective uptake of the antitumor drug 1-O-octadecyl-2-O-methyl-rac-glycero-3-phosphocholine (ET-18-OCH(3), Edelfosine) into lipid rafts of tumor cells, followed by its coaggregation with Fas death receptor (also known as APO-1 or CD95) and recruitment of apoptotic molecules into Fas-enriched rafts. Drug sensitivity was dependent on drug uptake and Fas expression, regardless of the presence of other major death receptors, such as tumor necrosis factor (TNF) receptor 1 or TNF-related apoptosis-inducing ligand R2/DR5 in the target cell. Drug microinjection experiments in Fas-deficient and Fas-transfected cells unable to incorporate exogenous ET-18-OCH(3) demonstrated that Fas was intracellularly activated. Partial deletion of the Fas intracellular domain prevented apoptosis. Unlike normal lymphocytes, leukemic T cells incorporated ET-18-OCH(3) into rafts coaggregating with Fas and underwent apoptosis. Fas-associated death domain protein, procaspase-8, procaspase-10, c-Jun amino-terminal kinase, and Bid were recruited into rafts, linking Fas and mitochondrial signaling routes. Clustering of rafts was necessary but not sufficient for ET-18-OCH(3)-mediated cell death, with Fas being required as the apoptosis trigger. ET-18-OCH(3)-mediated apoptosis did not require sphingomyelinase activation. Normal cells, including human and rat hepatocytes, did not incorporate ET-18-OCH(3) and were spared. This mechanism represents the first selective activation of Fas in tumor cells. Our data set a framework for the development of more targeted therapies leading to intracellular Fas activation and recruitment of downstream signaling molecules into Fas-enriched rafts.     

Efficacy of Participation-Focused Therapy on Performance of Physical Activity Participation Goals and Habitual Physical Activity in Children With Cerebral Palsy: A Randomized Controlled Trial

Objective

To determine the efficacy of a participation-focused therapy (ParticiPAte CP) on leisure-time physical activity goal performance and satisfaction and habitual physical activity (HPA) in children with CP.

扩阴器不同清洗消毒方法和流程改进的效果评价

目的:探讨本院消毒供应中心对扩阴器采用不同清洗消毒方法和流程改进的效果。方法:将2019年9月1日至2019年9月10日本院计生科人流包、上环包、妇科清宫包内扩阴器240只,随机均分成A、B、C三组,每组80只。A组采用常规多酶浸泡手工刷洗联合全自动清洗消毒器清洗消毒,B组采用95%乙醇擦洗多酶浸泡联合全自动清洗消毒器清洗消毒,C组采用碱性还原电位水刷洗联合全自动清洗消毒器清洗消毒。应用目测法、白纱布擦拭法、蛋白残留测试对比三组扩阴器清洗消毒效果和质量。结果:三组清洗效果均符合院感要求,C组目测法、白纱布擦拭法、蛋白残留测试检测值各项均显著优于A组、B组,差异均有统计学意义(P<0.05)。结论:用碱性还原电位水刷洗联合全自动清洗消毒器清洗消毒能有效去除扩阴器上的分泌物残留及碘伏,白纱布检测无污渍,切实可行。     

电针“百会”“神庭”对血管性痴呆大鼠学习记忆能力和海马突触结构与相关蛋白表达水平的影响

目的 观察电针“百会”“神庭”对血管性痴呆（vascular dementia,VD）大鼠学习和记忆功能的影响,并从突触结构及突触相关蛋白表达水平的角度揭示其作用机制。方法 将35只雄性SD大鼠随机分为假手术组、模型组、电针穴位组、电针非穴位组和奥拉西坦组,每组7只。采用改良双侧颈动脉结扎模型,电针穴位组大鼠选择“百会”“神庭”两穴治疗,电针非穴位组大鼠选择固定非穴位刺激,每次电针30 min,每日1次,连续干预14 d;奥拉西坦组大鼠选择腹腔注射奥拉西坦,50 mg/kg,每日1次,连续14 d。采用Morris水迷宫检测各组大鼠学习和空间记忆能力;透射电子显微镜观察各组大鼠海马CA1区突触结构;Western blot检测各组大鼠海马突触后致密蛋白95（postsynaptic density protein 95, PSD95）、GluA1、GluN2B和磷酸化GluN2B蛋白表达水平。结果 与假手术组比较,模型组大鼠学习期逃避潜伏时间延长,测试期跨越平台次数减少,目标象限停留时间显著缩短,大脑质量显著增加,海马CA1区突触结构数明显减少,海马PSD95、GluA1、GluN2B和磷酸化GluN2B蛋白表达水平均显著降低,差异均有统计学意义（P＜0.05）;与模型组比较,电针穴位组大鼠的学习期逃避潜伏时间缩短,测试期跨越平台次数增加,目标象限停留时间延长,大脑质量降低,CA1区突触结构数增多,海马PSD95、GluA1、GluN2B和磷酸化GluN2B蛋白表达水平增加,差异均有统计学意义（P＜0.05）。结论 电针“百会”“神庭”能改善VD大鼠的学习记忆功能,改变海马突触结构,分子机制可能和增加突触蛋白PSD95、GluA1和GluN2B的蛋白表达水平相关。     

lncRNA FAM95B1在胶质瘤中的表达及其影响LN382细胞增殖和迁移 的可能机制

目的：探讨lncRNA FAM95B1 对胶质瘤细胞增殖和迁移能力的影响并探究其相关作用机制。方法：选取2018年1月至2020年8月于合肥市第三人民医院行手术治疗的38例胶质瘤患者的胶质瘤组织及癌旁组织标本,利用qPCR检测胶质瘤组织与4种细胞系中FAM95B1的表达水平,以表达最低的胶质瘤LN382细胞为研究对象,转染空载质粒（对照组）或pcDNA3.1-FAM95B1质粒（实验组）。MTT法和划痕实验检测FAM95B1对LN382细胞增殖和迁移能力的影响。生物信息学分析技术和双荧光素酶基因报告实验预测并验证FAM95B1与miR-26a-5p及PTEN之间的相互作用机制,应用qPCR和WB法检测FAM95B1对miR-26a-5p和PTEN表达的影响。结果：FAM95B1在胶质瘤组织中的表达明显低于癌旁组织（P<0.01）。FAM95B1在多种胶质瘤细胞系中的表达均明显低于正常脑胶质细胞（均P<0.01）。过表达FAM95B1可以下调LN382细胞的增殖（P<0.05）和迁移能力（P<0.01）。FAM95B1 能够靶向结合 miR-26a-5p（P<0.01）,miR-26a-5p 能够靶向结合 PTEN mRNA（P<0.01）。过表达FAM95B1可下调LN382细胞中miR-26a-5p的表达（P<0.01）,促进PTEN mRNA的表达（P<0.01）。结论：在胶质瘤组织和细胞系中异常低表达的FAM95B1通过发挥竞争性内源RNA的功能抑制miR-26a-5p的表达而增强PTEN蛋白表达,抑制胶质瘤细胞系LN382的增殖和迁移。     

Can we increase the speed and efficacy of antidepressant treatments? Part II. Glutamatergic and RNA interference strategies

In the second part we focus on two treatment strategies that may overcome the main limitations of current antidepressant drugs. First, we review the experimental and clinical evidence supporting the use of glutamatergic drugs as fast-acting antidepressants. Secondly, we review the involvement of microRNAs (miRNAs) in the pathophysiology of major depressive disorder (MDD) and the use of small RNAs (e.g.., small interfering RNAs or siRNAs) to knockdown genes in monoaminergic and non-monoaminergic neurons and induce antidepressant-like responses in experimental animals.The development of glutamatergic agents is a promising venue for antidepressant drug development, given the antidepressant properties of the non-competitive NMDA receptor antagonist ketamine. Its unique properties appear to result from the activation of AMPA receptors by a metabolite [(2 S,6 S;2 R,6 R)-hydroxynorketamine (HNK)] and mTOR signaling. These effects increase synaptogenesis in prefrontal cortical pyramidal neurons and enhance serotonergic neurotransmission via descending inputs to the raphe nuclei. This view is supported by the cancellation of ketamine's antidepressant-like effects by inhibition of serotonin synthesis.We also review existing evidence supporting the involvement of miRNAs in MDD and the preclinical use of RNA interference (RNAi) strategies to target genes involved in antidepressant response. Many miRNAs have been associated to MDD, some of which e.g., miR-135 targets genes involved in antidepressant actions. Likewise, SSRI-conjugated siRNA evokes faster and/or more effective antidepressant-like responses. Intranasal application of sertraline-conjugated siRNAs directed to 5-HT_1A receptors and SERT evoked much faster changes of pre- and postsynaptic antidepressant markers than those produced by fluoxetine.     

阿魏酸钠对局灶性脑缺血突触后致密物质-95活化的影响

目的 :观察阿魏酸钠 (SF)对局灶性脑缺血损伤的保护作用及其对突触后致密物质 95 (PSD 95 )活化的影响。方法 :雄性 SD大鼠 4 6只 ,随机分成对照组和 SF组 (每组各 2 3只 ) ,分别于缺血时腹腔注射生理盐水 4 m l和 SF1 0 0 m g/ kg(用生理盐水 4 m l溶解 )。采用线栓法制备大脑中动脉阻闭 (MCAO)模型 ,1 6只大鼠(每组各 8只 )在脑缺血再灌注后 2 4 h神经行为学评分完成后被处死 ,取大脑行氯化三苯四唑 (TTC)染色以测量脑梗死容积 ;其他大鼠在脑缺血再灌注后的 30 m in、2 h、6 h、2 4 h和 72 h(各组每个时间点 3只 )分别被处死 ,取缺血侧和对侧皮质以及缺血侧纹状体行 Western blot分析。结果 :术后动物均存活。再灌注 2 4 h神经功能缺损评分对照组明显高于 SF组 ,SF组动物梗死容积〔(6 1 .5± 2 8.7) mm3〕明显小于对照组〔(1 6 8.1±4 2 .2 ) m m3,P<0 .0 1〕。 Western blot分析法表明 ,对照组再灌注 30 m in后 PSD 95在缺血皮质和纹状体活化显著增多并出现高峰 ,持续 72 h;在 SF治疗组 ,再灌注后 30 min PSD 95活化开始增多 ,高峰出现在 2 h,6 h后开始下降 ,而且 PSD 95活化在各时间点较对照组少 ,持续时间短。在对侧皮质未发现 PSD 95的活化。结论 :SF对局灶性脑缺血损伤有神经保护作用 ,并可减弱