BACKGROUND & AIMS: :A cDNA encoding a novel human extracellularly-regulated kinase (ERK) phosphatase, designated B59, was isolated from a B5/589 human mammary epithelial cell cDNA library. The 1104 nucleotide open reading frame encodes 368 amino acids including the highly conserved catalytic site sequence of protein phosphotyrosine phosphatases (PTPs), VXVHCXXGXXR, at amino acid position 276-287. The predicted 70 amino acid stretch surrounding the HC motif shares significant sequence identity with other human dual specificity PTPs (dsPTPs), including the known ERK PTPs CL100, PAC1, B23, as well as the dsPTPs VH-1 and VHR. B59 protein synthesized in vitro in a rabbit reticulocyte lysate dephosphorylated rat ERK1 and ERK2 proteins whose phosphorylation had been stimulated by v-mos kinase added to the lysate. Ectopic expression of B59 in NIH3T3 fibroblasts inhibited the induction of an oncogene-responsive promoter by the dominant-activating raf mutant, raf-BXB. Moreover, cotransfection of NIH3T3 cells with B59 inhibited morphological transformation by H-ras and v-raf oncogenes. These results suggest that B59 suppresses the transforming activity of H-ras or v-raf oncogenes through ERK dephosphorylation and inactivation.

背景与目标： ：从B5 / 589人乳腺上皮细胞cDNA文库中分离出一种编码新型人胞外调节激酶（ERK）磷酸酶的cDNA，命名为B59。 1104个核苷酸的开放阅读框编码368个氨基酸，其中包括高度保守的蛋白质磷酸酪氨酸磷酸酶（PTP）催化位点序列VXVHCXXGXXR，位于氨基酸位置276-287。 HC基序周围的预测的70个氨基酸段与其他人的双特异性PTP（dsPTP），包括已知的ERK PTP CL100，PAC1，B23以及dsPTP VH-1和VHR，具有明显的序列同一性。在兔网织红细胞裂解物中体外合成的B59蛋白将磷酸化的大鼠ERK1和ERK2蛋白磷酸化，并通过添加到裂解物中的v-mos激酶刺激了磷酸化。 B59在NIH3T3成纤维细胞中的异位表达抑制了由显性激活raf突变体raf-BXB诱导的癌基因应答性启动子。此外，NIH3T3细胞与B59的共转染可抑制H-ras和v-raf癌基因的形态转化。这些结果表明，B59通过ERK去磷酸化和失活抑制H-ras或v-raf癌基因的转化活性。

BACKGROUND & AIMS: :Human immunodeficiency virus (HIV) binds to cells via an interaction between CD4 and the virus envelope glycoprotein, gp120. Previous studies have localized the high affinity binding site for gp120 to the first domain of CD4, and monoclonal antibodies (mAbs) reactive with this region compete with gp120 binding and thereby block virus infectivity and syncytium formation. Despite a detailed understanding of the binding of gp120 to CD4, little is known of subsequent events leading to membrane fusion and virus entry. We describe two new mAbs reactive with the third domain of CD4 that inhibit steps subsequent to virus binding critical for HIV infectivity and cell fusion. Binding of recombinant gp120 or virus to CD4 is not inhibited by these antibodies, whereas infection and syncytium formation by a number of HIV isolates are blocked. These findings demonstrate that in addition to virus binding, CD4 may have an active role in membrane fusion.

背景与目标： 人类免疫缺陷病毒（HIV）通过CD4和病毒包膜糖蛋白gp120之间的相互作用与细胞结合。先前的研究已将gp120的高亲和力结合位点定位在CD4的第一个域，并且与该区域反应的单克隆抗体（mAb）与gp120结合竞争，从而阻断了病毒的感染性和合胞体的形成。尽管对gp120与CD4的结合有了详细的了解，但对导致膜融合和病毒进入的后续事件知之甚少。我们描述了与CD4的第三个域具有反应性的两个新的单克隆抗体，可抑制继病毒结合后对HIV感染性和细胞融合至关重要的步骤。重组gp120或病毒与CD4的结合不受这些抗体的抑制，而许多HIV分离株的感染和合胞体形成却被阻止。这些发现表明，除病毒结合外，CD4可能在膜融合中发挥积极作用。

BACKGROUND & AIMS: :In a context of automation of cryo-electron microscopy, we developed a novel method for improving visibility of diffraction rings in the power spectra of cryo-electron micrographs of vitreous ice (without carbon film or high concentration of diffracting material). We used these enhanced spectra to semi-automatically detect and remove micrographs and/or local areas introducing errors in the global 3D map (drifted and charged areas) or those unable to increase global signal-to-noise ratio (non-diffracting areas). Our strategy also allows a detection of micrographs/areas with a strong astigmatism. These images should be removed when using algorithms that do not correct astigmatism. Our sorting method is simple and fast since it uses the normalized cross-correlation between enhanced spectra and their copies rotated by 90 degrees. It owes its success mainly to the novel pre-processing of power spectra. The improved visibility also allows an easier visual check of accuracy of sorting. We show that our algorithm can even improve the visibility of diffraction rings of cryo-electron micrographs of pure water. Moreover, we show that this visibility depends strongly on ice thickness. This algorithm is implemented in the Xmipp (open-source image processing package) and is freely available for implementation in any other software package.

背景与目标： ：在低温电子显微镜自动化的背景下，我们开发了一种新颖的方法，用于改善玻璃冰（无碳膜或高浓度衍射材料）的低温电子显微镜照片的功率谱中衍射环的可见性。我们使用这些增强的光谱来半自动检测和移除显微照片和/或局部区域，从而在全局3D地图（漂移和带电区域）或无法增加全局信噪比（非衍射区域）的区域引入误差。我们的策略还允许检测具有强烈散光的显微照片/区域。当使用无法校正像散的算法时，应删除这些图像。我们的排序方法简单而快速，因为它使用了增强频谱与其旋转90度的副本之间的归一化互相关。它的成功主要归功于新颖的功率谱预处理。改进的可见性还可以使目视检查的准确性更加容易。我们证明了我们的算法甚至可以提高纯水冷冻电子显微照片的衍射环的可见性。此外，我们证明了这种能见度在很大程度上取决于冰的厚度。该算法在Xmipp（开源图像处理软件包）中实现，可免费用于任何其他软件包中。

BACKGROUND & AIMS: :Large-scale copy number variation that is cytogenetically visible in normal individuals has been described as euchromatic variation but needs to be distinguished from pathogenic euchromatic deletion or duplication. Here, we report eight patients (three families and two individuals) with interstitial deletions of 9q13-q21.12. Fluorescence in situ hybridisation with a large panel of BACs showed that all the deleted clones were from extensive tracts of segmentally duplicated euchromatin, copies of which map to both the long and short arms of chromosome 9. The variety of reasons for which these patients were ascertained, and the phenotypically normal parents, indicates that this is a novel euchromatic variant with no phenotypic effect. Further, four patients with classical euchromatic variants of 9q12/qh or 9p12 were also shown to have duplications or triplications of this segmentally duplicated material common to both 9p and 9q. The cytogenetic boundaries between the segmentally duplicated regions and flanking unique sequences were mapped to 9p13.1 in the short arm (BAC RP11-402N8 at 38.7 Mb) and to 9q21.12 in the long arm (BAC RP11-88I18 at 70.3 Mb). The BACs identified in this study should in future make it possible to differentiate between clinically significant deletions or duplications and euchromatic variants with no established phenotypic consequences.

背景与目标： ：在正常个体中在细胞遗传学上可见的大规模拷贝数变异已被描述为常染色体变异，但需要与致病性常染色体缺失或复制区分开。在这里，我们报告8例患者（3个家庭和2个个体）的间质缺失9q13-q21.12。与大量BAC的荧光原位杂交表明，所有缺失的克隆均来自节段重复的常染色质的广泛片段，其拷贝定位于9号染色体的长臂和短臂。确定这些患者的各种原因和表型正常的父母，表明这是一个新的常染色体变异，没有表型作用。此外，还显示了四名具有9q12 / qh或9p12的经典常染色体变体的患者具有这种重复出现或重复的9p12和9q共同的节段重复材料。片段重复区域和侧翼独特序列之间的细胞遗传学边界在短臂中定位为9p13.1（BAC RP11-402N8，38.7 Mb），在长臂中定位9q21.12，长臂（BAC RP11-88I18，70.3 Mb）。这项研究中确定的BAC将来应该可以区分具有临床意义的缺失或重复和常染色体变异，而没有确定的表型后果。

BACKGROUND & AIMS: :A major problem in the search for new cancer drug targets is that the drugs are often toxic to normal tissues and require high doses to kill tumor cells. Therefore cellular targets which appear to involve low dose responses to cancer therapy are especially interesting since they could selectively target normal tissues which are not targeted by the treatment and thus may be responsible for unpleasant side effects or may be amenable to exploitation in order to improve the therapeutic ratio. One such target, which is the subject of this review, is radiation-induced bystander effects [RIBE], which result in the observation of radiation like responses in cells which have not been irradiated. RIBE is a novel phenomenon which indicates that at low doses, cell signaling is more important than direct DNA damage. Historically, DNA has always been considered to be the target for radiation therapy. The growing realization that signaling is important opens up several important therapeutic strategies which will be discussed in this review. RIBE appears to be the result of a generalized stress response in tissues or cells which is expressed at the level of the tissue, organ or organism rather than at the level of the individual cell. The signals may be produced by all exposed cells, but the response may require a quorum of cells in order to be expressed. The major response involving low LET (x- or gamma-ray) radiation exposure discussed in the existing literature is a death response. This has many characteristics of apoptosis but may be detected in cell lines without p53 expression, although the death response is suppressed in many tumor cell lines. While a death response in unirradiated normal cells around a tumor might appear to be adverse, it can in fact be protective and remove damaged cells from the population. If harnessed correctly, it could lead to the development of new drugs aimed not at tissue destruction but at enabling homeostatic mechanisms to control tumor expansion. In this scenario, the level of harmful or beneficial response will be related to the background damage, carried by the cell population, and the genetic programme determining response to damage. This focus may be important when attempting to predict the consequences of mixed therapies involving radiation and other cytotoxic agents. In this review, our current knowledge of the mechanisms underlying the induction of bystander effects by ionizing radiation is reviewed, and the question of how bystander effects may be harnessed to produce a new generation of anti-cancer drugs aimed at stabilization of tissue homeostasis rather than tissue destruction is considered.

背景与目标： ：寻找新的癌症药物靶标的主要问题是该药物通常对正常组织有毒性，需要高剂量才能杀死肿瘤细胞。因此，细胞靶标似乎涉及对癌症治疗的低剂量反应，因此特别令人感兴趣，因为它们可以选择性地靶向未被治疗靶标的正常组织，因此可能引起令人不快的副作用，或者可能适于利用以改善治疗效果。治疗比率。辐射诱导的旁观者效应[RIBE]是本综述的主题之一，该效应导致在未辐射的细胞中观察到辐射样反应。 RIBE是一种新现象，表明在低剂量时，细胞信号传导比直接DNA损伤更为重要。从历史上看，DNA一直被认为是放射治疗的目标。人们日益认识到信号转导很重要，这开启了几种重要的治疗策略，本文将对此进行讨论。 RIBE似乎是组织或细胞中普遍的应激反应的结果，这种应激反应是在组织，器官或生物体的水平而不是单个细胞的水平表达的。信号可能由所有暴露的细胞产生，但响应可能需要一定数量的细胞才能表达。现有文献中讨论的涉及低LET（X射线或γ射线）辐射暴露的主要反应是死亡反应。这具有许多细胞凋亡特征，但尽管在许多肿瘤细胞系中死亡反应受到抑制，但在没有p53表达的细胞系中可能检测到。虽然在肿瘤周围未照射的正常细胞中的死亡反应似乎是不利的，但实际上可以起到保护作用，并从群体中清除受损的细胞。如果利用得当，它可能会导致开发新药物，其目的不是破坏组织，而是使稳态机制能够控制肿瘤的扩展。在这种情况下，有害或有益反应的水平将与细胞群体所携带的背景损伤以及决定对损伤的反应的遗传程序有关。当试图预测涉及放射线和其他细胞毒剂的混合疗法的后果时，这一重点可能很重要。在这篇综述中，我们对电离辐射诱发旁观者效应的潜在机制的现有知识进行了综述，并探讨了如何利用旁观者效应来生产旨在稳定组织稳态而不是稳定组织的新一代抗癌药物的问题。考虑组织破坏。

BACKGROUND & AIMS: :The pharmacological profile of MEN15596 or (6-methyl-benzo[b]thiophene-2-carboxylic acid [1-(2-phenyl-1R-{[1-(tetrahydropyran-4-ylmethyl)-piperidin-4-ylmethyl]-carbamoyl}-ethylcarbamoyl)-cyclopentyl]-amide), a novel potent and selective tachykinin NK2 receptor antagonist endowed with oral activity, is described. At the human recombinant tachykinin NK2 receptor, MEN15596 showed subnanomolar affinity (pKi 10.1) and potently antagonized (pKB 9.1) the neurokinin A-induced intracellular calcium release. MEN15596 selectivity for the tachykinin NK2 receptor was assessed by binding studies at the recombinant tachykinin NK1 (pKi 6.1) and NK3 (pKi 6.4) receptors, and at a number of 34 molecular targets including receptors, transporters and ion channels. In isolated smooth muscle preparations MEN15596 showed a marked species selectivity at the tachykinin NK2 receptor with the highest antagonist potency in guinea-pig colon, human and pig bladder (pKB 9.3, 9.2 and 8.8, respectively) whereas it was three orders of magnitude less potent in the rat and mouse urinary bladder (pKB 6.3 and 5.8, respectively). In agreement with binding experiments, MEN15596 showed low potency in blocking selective NK1 or NK3 receptor agonist-induced contractions of guinea-pig ileum preparations (pA2

BACKGROUND & AIMS: :The human iodotyrosine dehalogenase 1 (DEHAL1) gene is composed of six exons. Two isoforms (DEHAL1 and DEHAL1B) have been published in GenBank, both of which have a nitroreductase domain and arise from differential splicing in exon 5. We recently showed that the DEHAL1 isoform is a transmembrane protein that efficiently catalyzes the NADPH-dependent deiodination of mono (L-MIT) and diiodotyrosine (L-DIT) in human embryonic kidney-293 (HEK293) cells. In the present study, we establish the existence of a new transcript, DEHAL1C, in the human thyroid with a terminal exon that lacks in the DEHAL1 transcript. This exon is the complete exon 5, which is spliced in the DEHAL1B mRNA variant. These two variants encode proteins with differing C-terminal domains. Using quantitative reverse transcription polymerase chain reaction, we found that the expression of the mRNA of DEHAL1C and DEHAL1B was lower than that of DEHAL1 mRNA in the thyroid. We also observed that human DEHAL1B and DEHAL1C proteins are rapidly degraded in stably transfected HEK293 cells, unlike the DEHAL1 protein, and that exposure to the proteasome inhibitor MG132 resulted in accumulation of these proteins that was markedly time- and concentration-dependent. These findings show that the cytoplasmic tail could play a role in the stability of the protein.

背景与目标： ：人碘酪氨酸脱卤素酶1（DEHAL1）基因由六个外显子组成。 GenBank中已公开了两种同工型（DEHAL1和DEHAL1B），它们均具有硝基还原酶结构域，并来自外显子5的差异剪接。 （L-MIT）和二碘代酪氨酸（L-DIT）在人类胚胎肾293（HEK293）细胞中。在本研究中，我们在人甲状腺中建立了一个新的转录物DEHAL1C的存在，其末端外显子缺少DEHAL1转录物。该外显子是完整的外显子5，其剪接在DEHAL1B mRNA变异体中。这两个变体编码具有不同C末端结构域的蛋白质。通过定量逆转录聚合酶链反应，我们发现甲状腺中DEHAL1C和DEHAL1B的mRNA表达低于DEHAL1 mRNA的表达。我们还观察到，与DEHAL1蛋白不同，人DEHAL1B和DEHAL1C蛋白在稳定转染的HEK293细胞中迅速降解，并且暴露于蛋白酶体抑制剂MG132导致这些蛋白的积累具有明显的时间和浓度依赖性。这些发现表明，胞质尾部可以在蛋白质的稳定性中起作用。

BACKGROUND & AIMS: :Inborn errors of vitamin B(12) (cobalamin) metabolism are characterized by decreased production of active cobalamin cofactors and subsequent deficiencies in the activities of methionine synthase and methylmalonyl-CoA mutase. With the recent discovery of the cblJ defect in two patients with phenotypes mimicking the cblF defect, there are nine genes known to be involved in cobalamin metabolism. The new defect is caused by mutations in the ABCD4 gene, encoding an ABC transporter. At the moment, there is no clear distinction between the cblJ and cblF defects either clinically or biochemically, and both defects result in blocks in the transport of cobalamin from the lysosome to the cytoplasm. A patient was diagnosed with hyperhomocysteinemia and methylmalonic aciduria at the age of 8 years. Incorporations of both [(14)C]propionate and [(14)C]methyltetrahydrofolate in cultured fibroblasts were within reference ranges and thus too high to allow for complementation analysis. We observed decreased synthesis of both adenosylcobalamin and methylcobalamin and accumulation of unmetabolized cyanocobalamin. Exome sequencing was performed to identify causative mutation(s) and Sanger re-sequencing was performed to validate segregation of mutation in the family. By this approach, a homozygous mutation, c.423C>G, in the ABCD4 gene was identified. Here, we report the successful application of exome sequencing for diagnosis of a rare inborn error of vitamin B(12) metabolism in a patient whose unusual presentation precluded diagnosis using standard biochemical and genetic approaches. The patient represents only the third known patient with the cblJ disorder.

背景与目标： ：维生素B（12）（钴胺素）代谢的先天性错误的特征在于活性钴胺素辅因子的产生减少以及蛋氨酸合酶和甲基丙二酰辅酶A突变酶的活性随后不足。随着最近在两名模仿cblF缺陷的表型患者中发现cblJ缺陷，已知有9个基因与钴胺素代谢有关。新的缺陷是由编码ABC转运蛋白的ABCD4基因突变引起的。目前，在临床或生化方面，cblJ和cblF缺陷之间尚无明确区分，并且两种缺陷均导致钴胺素从溶酶体到细胞质的转运受阻。一名患者在8岁时被诊断出患有高同型半胱氨酸血症和甲基丙二酸尿症。 [（14）C]丙酸酯和[（14）C]甲基四氢叶酸在培养的成纤维细胞中的掺入均在参考范围内，因此含量过高，无法进行互补分析。我们观察到腺苷钴胺素和甲基钴胺素的合成减少以及未代谢的氰钴胺素的积累。进行了外显子组测序以鉴定致病突变，并进行了桑格重测序以验证家族中突变的分离。通过这种方法，鉴定出ABCD4基因中的纯合突变，即c.423C> G。在这里，我们报告外显子组测序在维生素B（12）代谢的罕见先天性错误的诊断中成功应用，该患者的异常表现排除了使用标准生化和遗传方法进行诊断的可能性。该患者仅代表第三位已知的cblJ疾病患者。

BACKGROUND & AIMS: :Silent information regulator 2 (Sir2) enzymes or sirtuins are a family of NAD(+)-dependent protein N(ε)-acetyl-lysine (AcK) deacetylases. Sirtuins are also evolutionarily conserved proteins that are present in all kingdoms of life ranging from bacteria to humans. Interestingly, it was recently found that the sirtuins found in various human parasites (especially the Plasmodium, Trypanosoma, and Leishmania species) were pro-survival for the parasites under various conditions. Therefore, these parasitic sirtuins have emerged as novel anti-parasitic therapeutic targets. This article reviews the currently available structural, biochemical, pharmacological, and medicinal chemistry studies on these enzymes, and discusses the perspectives of selectively targeting the parasitic sirtuins as a novel therapeutic strategy for the human parasitic diseases.

背景与目标： ：沉默信息调节剂2（Sir2）酶或sirtuins是NAD（）依赖性蛋白N（ε）-乙酰赖氨酸（AcK）脱乙酰基酶的一个家族。 Sirtuins还是进化上保守的蛋白质，存在于从细菌到人类的所有生命王国中。有趣的是，最近发现在各种人类寄生虫（尤其是疟原虫，锥虫和利什曼原虫）中发现的沉默调节蛋白在各种条件下均可存活。因此，这些寄生沉默调节蛋白已经成为新型的抗寄生虫治疗靶标。本文回顾了有关这些酶的当前可用的结构，生化，药理和药物化学研究，并讨论了选择性靶向寄生虫沉默调节蛋白作为人类寄生虫疾病的新型治疗策略的观点。

BACKGROUND & AIMS: :Alkenylboronic acids have shown important biological activities that contribute to neuroprotection. We have determined their influence on the β-amyloid (βA) aggregation process, β-secretase and acethylcholinesterase activities on cell-free systems, on the redox and lipid peroxidation status, and on the vulnerability to apoptotic death in an APPswe neuroblastoma cell line, before and after hydrogen peroxide treatment. We have discovered that 2-arylvinylboronic acids and some of their esters possess a set of properties which makes them highly useful as neuroprotective agents affecting multiple biological targets involved in AD. These properties are not paralleled by the related 2-arylboronic acids.

背景与目标： 烯基硼酸已显示出重要的生物活性，可促进神经保护作用。我们已经确定了它们对β-淀粉样蛋白（βA）聚集过程，β-分泌酶和乙酰胆碱酯酶活性对无细胞系统，氧化还原和脂质过氧化状态的影响，以及对APPswe神经母细胞瘤细胞株凋亡死亡的脆弱性的影响，过氧化氢处理前后。我们已经发现2-芳基乙烯基硼酸及其某些酯具有一系列特性，这使得它们非常有用地用作影响涉及AD的多个生物学靶标的神经保护剂。这些性能是相关的2-芳基硼酸无法比拟的。

BACKGROUND & AIMS: :Novel double-capped triplet drugs, which have one pharmacophore unit and two epoxymethano or dimethylepoxymethano structures (termed cap or diMe-cap structures, respectively) were synthesized. Key intermediate oxazoline 16 derived from acetone enabled the effective synthesis of double-capped triplets. SYK-134 (7a) and SYK-135 (8a) with N-cyclopropylmethyl substituent and cap structures showed selectivities for the κ opioid receptor. On the other hand, the N-Me series exhibited selectivities for the μ opioid receptor. The double-capped triplet drugs with diMe-cap structures preferred the μ receptor independently of their N-substituents. SYK-385 (19b), one of the μ-selective double-capped triplet drugs, showed the highest selectivity for the μ receptor among the reported μ-selective nonpeptide ligands.

背景与目标： 合成了具有一个药效团单元和两个环氧亚甲基或二甲基环氧亚甲基结构（分别称为帽或二甲基-帽结构）的新型双封端三联体药物。源自丙酮的关键中间体恶唑啉16能够有效合成双封三胞胎。具有N-环丙基甲基取代基和帽结构的SYK-134（7a）和SYK-135（8a）对κ阿片受体具有选择性。另一方面，N-Me系列对μ阿片受体具有选择性。具有diMe-cap结构的双封端三联体药物优选μ受体，而与它们的N取代基无关。微米选择性双封端三联体药物之一SYK-385（19b）在已报道的微米选择性非肽配体中显示出对微米受体的最高选择性。

BACKGROUND & AIMS: :Nuclear protein in testis (NUT)-midline carcinoma (NMC) is a rare, aggressive disease typically presenting with a single t(15;19) translocation that results in the generation of a bromodomain-containing protein 4 (BRD4)-NUT fusion. PER-624 is a cell line generated from an NMC patient with an unusually complex karyotype that gave no initial indication of the involvement of the NUT locus. Analysis of PER-624 next-generation transcriptome sequencing (RNA-Seq) using the algorithm FusionFinder identified a novel transcript in which Exon 15 of BRD4 was fused to Exon 2 of NUT, therefore differing from all published NMC fusion transcripts. The three additional exons contained in the PER-624 fusion encode a series of polyproline repeats, with one predicted to form a helix. In the NMC cell line PER-403, we identified the 'standard' NMC fusion and two novel isoforms. Knockdown by small interfering RNA in either cell line resulted in decreased proliferation, increased cell size and expression of cytokeratins consistent with epithelial differentiation. These data demonstrate that the novel BRD4-NUT fusion in PER-624 encodes a functional protein that is central to the oncogenic mechanism in these cells. Genomic PCR indicated that in both PER-624 and PER-403, the translocation fuses an intron of BRD4 to a region upstream of the NUT coding sequence. Thus, the generation of BRD4-NUT fusion transcripts through post-translocation RNA-splicing appears to be a common feature of these carcinomas that has not previously been appreciated, with the mechanism facilitating the expression of alternative isoforms of the fusion. Finally, ectopic expression of wild-type NUT, a protein normally restricted to the testis, could be demonstrated in PER-403, indicating additional pathways for aberrant cell signaling in NMC. This study contributes to our understanding of the genetic diversity of NMC, an important step towards finding therapeutic targets for a disease that is refractory to current treatments.

背景与目标： ：睾丸（NUT）-中线癌（NMC）中的核蛋白是一种罕见的侵袭性疾病，通常表现为单个t（15; 19）易位，导致含溴结构域的蛋白4（BRD4）-NUT融合的产生。 PER-624是由具有异常复杂核型的NMC患者产生的细胞系，未初步显示NUT基因座的参与。使用FusionFinder算法对PER-624下一代转录组测序（RNA-Seq）进行分析，发现了一种新颖的转录本，其中BRD4的第15外显子与NUT的第2外显子融合，因此不同于所有已发表的NMC融合转录本。 PER-624融合物中包含的三个附加外显子编码一系列聚脯氨酸重复序列，其中一个被预测会形成螺旋。在NMC细胞系PER-403中，我们确定了“标准” NMC融合蛋白和两种新型同工型。在任一细胞系中通过小分子干扰RNA敲低导致增殖减少，细胞大小增加和与上皮分化一致的细胞角蛋白表达。这些数据表明，PER-624中的新型BRD4-NUT融合蛋白编码的功能蛋白是这些细胞中致癌机制的核心。基因组PCR表明，在PER-624和PER-403中，易位将BRD4的内含子融合到NUT编码序列上游的区域。因此，通过易位后RNA剪接产生BRD4-NUT融合转录本似乎是这些癌的一个共同特征，以前尚未被人们认识到，其机制促进了融合的其他亚型的表达。最后，可以在PER-403中证实野生型NUT（通常限于睾丸的蛋白质）的异位表达，这表明NMC中异常细胞信号传导的其他途径。这项研究有助于我们了解NMC的遗传多样性，这是为目前治疗难以治疗的疾病寻找治疗靶标的重要一步。

BACKGROUND & AIMS: OBJECTIVES:Pancreatic endocrine tumors (PETs) share numerous features with gastrointestinal neuroendocrine (carcinoid) tumors. Targets of novel therapeutic strategies previously assessed in carcinoid tumors were analyzed in PETs (44 cases). METHODS:Activating mutations in EGFR, KIT, and PDGFRA and nonresponse mutations in KRAS were evaluated. Copy number of EGFR and HER-2/neu was quantified by fluorescence in situ hybridization. Expression of EGFR, PDGFRA, VEGFR1, TGFBR1, Hsp90, SSTR2A, SSTR5, IGF1R, mTOR, and MGMT was measured immunohistochemically. RESULTS:Elevated EGFR copy number was found in 38% of cases but no KRAS nonresponse mutations. VEGFR1, TGFBR1, PDGFRA, SSTR5, SSTR2A, and IGF1R exhibited the highest levels of expression in the largest percentages of PETs.Anticancer drugs BMS-754807 (selective for IGF1R/IR), 17-(allylamino)-17-demethoxygeldanamycin (17-AAG, targeting Hsp90), and axitinib (directed toward VEGFR1-3/PDGFRA-B/KIT) induced growth inhibition of human QGP-1 PET cells with IC50 values (nM) of 273, 723, and 743, respectively. At growth-inhibiting concentrations, BMS-754807 inhibited IGF1R phosphorylation; 17-AAG induced loss of EGFR, IGF1R, and VEGFR2; and axitinib increased p21(CDKN1A) expression without inhibiting VEGFR2 phosphorylation. CONCLUSIONS:Results encourage further research into multidrug strategies incorporating inhibitors targeting IGF1R or Hsp90 and into studies of axitinib combined with conventional chemotherapeutics toxic to tumor cells in persistent growth arrest.

背景与目标： 目的：胰腺内分泌肿瘤（PET）与胃肠道神经内分泌（类癌）肿瘤具有许多特征。以前在类癌肿瘤中评估过的新治疗策略的靶标已在PET（44例）中进行了分析。
方法：评估EGFR，KIT和PDGFRA的激活突变以及KRAS的无应答突变。通过荧光原位杂交定量EGFR和HER-2 / neu的拷贝数。免疫组织化学法检测EGFR，PDGFRA，VEGFR1，TGFBR1，Hsp90，SSTR2A，SSTR5，IGF1R，mTOR和MGMT的表达。
结果：38％的病例发现EGFR拷贝数升高，但没有KRAS无反应突变。 VEGFR1，TGFBR1，PDGFRA，SSTR5，SSTR2A和IGF1R在最大百分比的PET中表现出最高的表达水平。抗癌药BMS-754807（对IGF1R / IR选择性），17-（烯丙胺基）-17-去甲氧基格尔德霉素（17-靶向Hsp90的AAG和axitinib（针对VEGFR1-3 / PDGFRA-B / KIT）诱导人QGP-1 PET细胞的生长抑制，IC50值（nM）分别为273、723和743。在抑制生长的浓度下，BMS-754807抑制了IGF1R磷酸化。 17-AAG诱导的EGFR，IGF1R和VEGFR2丢失；阿昔替尼在不抑制VEGFR2磷酸化的情况下增加p21（CDKN1A）的表达。
结论：结果鼓励对结合靶向IGF1R或Hsp90的抑制剂的多药策略和阿昔替尼与常规化学治疗药物联合治疗对持续生长停滞有毒的肿瘤细胞进行进一步研究。

BACKGROUND & AIMS: BACKGROUND:There is conflicting evidence as to whether achievement of cholesterol targets at the population level is dependent on the choice and cost of statin. AIM:To investigate the practice-level relationship between cholesterol quality indicators in patients with heart disease, stroke, and diabetes and prescribing of low-cost statins. DESIGN AND SETTING:Correlations and linear regression modelling of retrospective cross-sectional practice-level data with potential explanatory variables in 7909 (96.4%) general practices in England in 2008-2009. METHOD:Quality indicator data were obtained from the Information Centre and prescribing data from the NHS Business Authority. A 'cholesterol quality indicator' score was constructed by dividing the numbers of patients achieving the target for cholesterol control of ≤5 mmol/l in stroke, diabetes, and heart disease by the numbers on each register. A 'low-cost statin' ratio score was constructed by dividing the numbers of defined daily doses of simvastatin and pravastatin by the total numbers of defined daily doses of statins. RESULTS:Simvastatin accounted for 83.3% (standard deviation [SD] = 15.7%) of low-cost statins prescribed and atorvastatin accounted for 85.7% (SD = 14.8%) of high-cost statins prescribed. The mean cholesterol score was 73.7% (SD = 6.0%). Practices using a higher proportion of the low-cost statins were less successful in achieving cholesterol targets. An increase of 10% in the prescribing of low-cost statins was associated with a decrease of 0.46% in the cholesterol quality indicator score (95% confidence interval = -0.54% to -0.38%, P<0.001). CONCLUSION:Greater use of low-cost statins was associated with a small reduction in cholesterol control.

背景与目标： 背景：关于在人群水平上实现胆固醇目标是否取决于他汀类药物的选择和成本，有相互矛盾的证据。
目的：探讨心脏病，中风和糖尿病患者的胆固醇质量指标与廉价他汀类药物处方之间的实践水平关系。
设计与设置：回顾性横截面实践水平数据的相关性和线性回归模型，该数据具有2008年至2009年英格兰7909例（96.4％）常规实践中的潜在解释变量。
方法：质量指标数据是从信息中心获得的，处方数据是从NHS商业管理局获得的。通过将中风，糖尿病和心脏病中达到胆固醇控制目标≤5mmol / l的患者人数除以每个登记册上的人数，来构建“胆固醇质量指标”评分。通过将辛伐他汀和普伐他汀的每日定义剂量数除以他汀类药物的每日定义总数，可以构建“低成本他汀类药物”比率评分。
结果：辛伐他汀占处方中低成本他汀类药物的83.3％（标准偏差[SD] = 15.7％），阿托伐他汀占处方中低成本他汀类药物的85.7％（SD = 14.8％）。平均胆固醇得分为73.7％（SD = 6.0％）。使用较高比例的低成本他汀类药物的做法在实现胆固醇目标方面不太成功。低成本他汀类药物处方的增加10％与胆固醇质量指标得分的0.46％的降低相关（95％置信区间= -0.54％至-0.38％，P <0.001）。
结论：大量使用低成本他汀类药物与降低胆固醇控制量有关。

BACKGROUND & AIMS: :Doxazosin used in benign prostatic hyperplasia has the side effects of causing hypotension and the risk of heart failure. The 3 targets of α(1A)-adrenoceptors (in the prostate), α(1D)-adrenoceptors (in the aorta), and an unknown mechanism (in the heart) are involved, respectively. We hypothesized that there is a chiral recognition of doxazosin enantiomers in the 3 targets. Using isolated rat aorta (α(1D)-adrenoceptors) and rabbit prostate (α(1A)-adrenoceptors), we examined pA(2) and pK(B) values of doxazosin enantiomers. We observed chronotropic and inotropic effects of doxazosin enantiomers in isolated rat and rabbit heart tissues. (-)Doxazosin and (+)doxazosin produced a shift to the right of concentration-contraction curves for noradrenalin (aorta) and phenylephrine (prostate smooth muscle). The pA(2) value of (-)doxazosin (8.625 ± 0.053) was smaller than (+)doxazosin (9.503 ± 0.051) in rat aorta, but their pK(B) values in rabbit prostate were the same. In rat and rabbit heart tissues, (+)doxazosin (3-30 µmol·L(-1)) significantly decreased atrial rate, and produced negative inotropic effects; however, (-)doxazosin did not affect the atrial rate, and produced positive inotropic effects in the atria. Thus, the chiral carbon atom of doxazosin does not affect its activity at the therapeutic target of α(1A)-adrenoceptors in the prostate, but significantly changes its blocking activity against α(1D)-adrenoceptors in the aorta, and produces opposite inotropic effects in the atria via an α(1)-adrenoceptor-independent mechanism.

背景与目标： ：多沙唑嗪用于前列腺增生症具有引起低血压和心力衰竭风险的副作用。 α（1A）-肾上腺素受体（在前列腺中），α（1D）-肾上腺素受体（在主动脉中）和未知机制（在心脏中）的三个靶标分别涉及。我们假设在3个靶标中有手性识别多沙唑嗪对映体。使用分离的大鼠主动脉（α（1D）-肾上腺素受体）和兔前列腺（α（1A）-肾上腺素受体），我们检查了多沙唑嗪对映体的pA（2）和pK（B）值​​。我们观察到了多沙唑嗪对映异构体在离体大鼠和兔心脏组织中的变时性和变力作用。 （-）多沙唑嗪和（）多沙唑嗪使去甲肾上腺素（主动脉）和去氧肾上腺素（前列腺平滑肌）的浓度-收缩曲线向右移动。大鼠主动脉中（-）恶唑烷的pA（2）值（8.625±0.053）小于（）恶唑烷（9.503±0.051），但它们在兔前列腺中的pK（B）值​​相同。在大鼠和兔子的心脏组织中，（）多沙唑嗪（3-30 µmol·L（-1））显着降低心房率，并产生负性变力作用；然而，（-）doxazosin不会影响心房率，并在心房产生正性肌力作用。因此，多沙唑嗪的手性碳原子不影响其对前列腺中α（1A）-肾上腺素受体的治疗靶点的活性，但会显着改变其对主动脉中α（1D）-肾上腺素受体的阻断活性，并产生相反的正性肌力作用通过独立于α（1）-肾上腺素受体机制的心房。