BACKGROUND & AIMS: :We have previously shown that the interaction of Ca2+/calmodulin with the metabotropic glutamate receptor type 7 (mGluR7) promotes the G-protein-mediated inhibition of voltage-sensitive Ca2+ channels (VSCCs) seen upon agonist activation. Here, we performed a yeast two-hybrid screen of a new-born rat brain cDNA library using the cytoplasmic C-terminal tail of mGluR7 as bait and identified macrophage myristoylated alanine-rich c-kinase substrate (MacMARCKS) as a binding protein. The interaction was confirmed in vitro and in vivo by pull-down assays, immunoprecipitation, and colocalization of mGluR7 and MacMARCKS in transfected HEK293 cells and cultured cerebellar granule cells. Binding of MacMARCKS to mGluR7 was antagonized by Ca2+/calmodulin. In neurons, cotransfection of MacMARCKS with mGluR7, but not mGluR7 mutants unable to bind MacMARCKS, reduced the G-protein-mediated tonic inhibition of VSCCs in the absence of mGluR7 agonist. These results suggest that competitive interactions of Ca2+/calmodulin and MacMARCKS with mGluR7 control the tonic inhibition of VSCCs by G-proteins.

背景与目标： ：我们之前已经证明，Ca2 /钙调蛋白与7型代谢型谷氨酸受体（mGluR7）的相互作用促进了激动剂激活后G蛋白介导的对电压敏感Ca2通道（VSCC）的抑制。在这里，我们使用mGluR7的胞质C末端尾巴作为诱饵，对新生大鼠大脑cDNA文库进行了酵母双杂交筛选，并鉴定了巨噬细胞肉豆蔻酰化的富含丙氨酸的c激酶底物（MacMARCKS）作为结合蛋白。通过下拉测定，免疫沉淀以及mGluR7和MacMARCKS在转染的HEK293细胞和培养的小脑颗粒细胞中的共定位，在体外和体内证实了这种相互作用。 Ca2 /钙调蛋白可拮抗MacMARCKS与mGluR7的结合。在神经元中，将MacMARCKS与mGluR7共转染，但不能与不能结合MacMARCKS的mGluR7突变体共转染，可在不存在mGluR7激动剂的情况下减少VSCC的G蛋白介导的强直抑制。这些结果表明Ca2 /钙调蛋白和MacMARCKS与mGluR7的竞争性相互作用控制了G蛋白对VSCC的强直抑制。

BACKGROUND & AIMS: The amino-terminal segment of the membrane-anchored subunit of influenza hemagglutinin (HA) plays a crucial role in membrane fusion and, hence, has been termed the fusion peptide. We have studied the secondary structure, orientation, and effects on the bilayer structure of synthetic peptides corresponding to the wild-type and several fusogenic and nonfusogenic mutants with altered N-termini of the influenza HA fusion peptide by fluorescence, circular dichroism, and Fourier transform infrared spectroscopy. All peptides contained segments of alpha-helical and beta-strand conformation. In the wild-type fusion peptide, 40% of all residues were in alpha-secondary and 30% in beta-secondary structures. By comparison, the nonfusogenic peptides exhibited larger beta/alpha secondary structure ratios. The order parameters of the helices and the amide carbonyl groups of the beta-strands of the wild-type fusion peptide were measured separately, based on the infrared dichroism of the respective absorption bands. Order parameters in the range 0.1-0.7 were found for both segments of the wild-type peptide, which indicates that they are most likely aligned at oblique angles to the membrane normal. The nonfusogenic but not the fusogenic peptides induced splitting of the infrared absorption band at 1735 cm(-1), which is assigned to stretching vibrations of the lipid ester carbonyl bond. This splitting, which reports on an alteration of the hydrogen bonds formed between the lipid ester carbonyls and water and/or hydrogen-donating groups of the fusion peptides, correlated with the beta/alpha ratio of the peptides, suggesting that unpaired beta-strands may replace water molecules and hydrogen-bond to the lipid ester carbonyl groups. The profound structural changes induced by single amino acid replacements at the extreme N-terminus of the fusion peptide further suggest that tertiary or quaternary structural interactions may be important when fusion peptides bind to lipid bilayers.

背景与目标： 流感血凝素（HA）的膜锚定亚基的氨基末端片段在膜融合中起着至关重要的作用，因此被称为融合肽。我们已经通过荧光，圆二色性和傅里叶变换研究了对应于野生型和几个融合和非融合突变体的流感病毒HA融合肽的N末端改变的二级肽的二级结构，方向及其对双层肽结构的影响。红外光谱。所有肽都包含α-螺旋和β-链构象的区段。在野生型融合肽中，所有残基的40％在α-二级结构中，而30％在β-二级结构中。相比之下，非融合肽表现出较大的β/α二级结构比。基于各个吸收带的红外二色性，分别测量野生型融合肽的β链的螺旋和酰胺羰基的有序参数。对于野生型肽的两个片段，发现有序参数在0.1-0.7范围内，这表明它们最有可能以与膜法线倾斜的角度排列。非融合肽而非融合肽诱导了1735 cm（-1）处的红外吸收带的分裂，这被分配给了脂质酯羰基键的拉伸振动。该分裂报告了脂质酯羰基与融合肽的水和/或供氢基团之间形成的氢键的改变，与肽的β/α比率相关，表明未配对的β链可能取代水分子，并与脂质酯的羰基氢键合。融合肽极端N端由单个氨基酸置换引起的深刻结构变化进一步表明，当融合肽结合脂质双层时，三级或四级结构相互作用可能很重要。

BACKGROUND & AIMS: :Receptor tyrosine kinases (RTKs) are critical for normal cell growth, differentiation, and development, but they contribute to various pathological conditions when disrupted. Activation of RTKs stimulates a plethora of pathways, including the ubiquitylation and endocytosis of the receptor itself. Although endocytosis terminates RTK signaling, it has emerged as a requisite step in RTK activation of signaling pathways. We have discovered that the endocytic scaffolding protein intersectin (ITSN) cooperated with epidermal growth factor receptor (EGFR) in the regulation of cell growth and signaling. However, a biochemical link between ITSN and EGFR was not defined. In this study, we demonstrate that ITSN is a scaffold for the E3 ubiquitin ligase Cbl. ITSN forms a complex with Cbl in vivo mediated by the Src homology (SH) 3 domains binding to the Pro-rich COOH terminus of Cbl. This interaction stimulates the ubiquitylation and degradation of the activated EGFR. Furthermore, silencing ITSN by RNA interference attenuated EGFR internalization as well as activation of the extracellular signal-regulated kinasemitogen-activated protein kinase pathway, thereby demonstrating the importance of ITSN in EGFR function. Given the cooperativity between ITSN and additional RTKs, these results point to an important evolutionarily conserved, regulatory role for ITSN in RTK function that is necessary for both signaling from receptors as well as the ultimate termination of receptor signaling.

背景与目标： 受体酪氨酸激酶（RTKs）对于正常细胞的生长，分化和发育至关重要，但是当它们被破坏时，它们会导致各种病理状况。 RTK的激活刺激了许多途径，包括受体自身的泛素化和内吞作用。尽管内吞作用终止了RTK信号传导，但它已成为RTK激活信号通路的必要步骤。我们已经发现，内吞支架蛋白intersectin（ITSN）与表皮生长因子受体（EGFR）协同调节细胞生长和信号传导。但是，尚未定义ITSN和EGFR之间的生化联系。在这项研究中，我们证明了ITSN是E3泛素连接酶Cbl的支架。 ITSN在体内与Cbl形成复合物，该复合物是通过与Cbl富含Pro的COOH末端结合的Src同源性（SH）3结构域介导的。这种相互作用刺激了活化的EGFR的泛素化和降解。此外，通过RNA干扰使ITSN沉默可以减弱EGFR的内在化以及细胞外信号调节激酶有丝分裂原激活的蛋白激酶途径的激活，从而证明了ITSN在EGFR功能中的重要性。考虑到ITSN和其他RTK之间的协作性，这些结果表明ITSN在RTK功能中具有重要的进化保守的调节作用，这对于受体的信号传导以及受体信号传导的最终终止都是必需的。

BACKGROUND & AIMS: The exact mechanism for the decrease in intestinal calcium absorption with age is not yet understood. A decrease with age in serum 1,25-dihydroxyvitamin D (1,25(OH)2D) or a decrease in the intestinal vitamin D receptor (VDR) protein concentration are possible causes. The objective of this study was to examine the effect of age on these factors. Fifty-nine young women age 25-35 years were compared with 41 elderly women age 65-83 years who underwent measurements of VDR, calcium absorption using a 20 mg and 100 mg calcium carrier, and calciotropic hormones. Calcium absorption by both tests was lower in the elderly women compared with the young women (p < 0.05). Serum 1,25(OH)2D and duodenal VDR protein concentration were not significantly different between the two age groups. Serum 1,25(OH)2D correlated with the 20 mg calcium absorption test in both young (r = 0.35, p < 0.007) and elderly women (r = 0.58, p < 0.0001) and with the 100 mg calcium absorption in the elderly (r = 0.32; p < 0.05). VDR did not correlate with calcium absorption in young women or elderly women, nor did VDR correlate with serum 1,25(OH)2D and serum 25-hydroxyvitamin D. In summary, the decrease in calcium absorption cannot be explained by a decrease in intestinal VDR. The correlation between serum 1,25(OH)2D and both calcium absorption tests only accounts for 12-30% of the variance in the age-related change in the calcium absorption tests. Other factors, not yet understood, are responsible for the decline in calcium absorption with age.

背景与目标： 随着年龄的增长，肠道钙吸收减少的确切机制尚不清楚。血清1,25-二羟基维生素D（1,25（OH）2D）随年龄的减少或肠道维生素D受体（VDR）蛋白质浓度的减少是可能的原因。这项研究的目的是研究年龄对这些因素的影响。比较了59名年龄在25-35岁之间的年轻女性与41位年龄在65-83岁之间的女性，这些女性进行了VDR测量，使用20 mg和100 mg钙载体的钙吸收量以及亲钙性激素。与年轻女性相比，老年女性的两种测试中的钙吸收均较低（p <0.05）。在两个年龄组之间，血清1,25（OH）2D和十二指肠VDR蛋白浓度无显着差异。血清1,25（OH）2D与年轻（r = 0.35，p <0.007）和老年妇女（r = 0.58，p <0.0001）的20 mg钙吸收测试以及老年人的100 mg钙吸收相关（r ＝ 0.32； p ＜0.05）。 VDR与年轻妇女或老年妇女的钙吸收无关，也不与血清1,25（OH）2D和血清25-羟维生素D相关。 VDR。血清1,25（OH）2D与两种钙吸收试验之间的相关性仅占钙吸收试验中与年龄相关的变化方差的12％至30％。钙吸收随着年龄的增长而下降的其他原因尚不明确。

BACKGROUND & AIMS: :Several molecular-targeted therapeutics have been tested in clinical trials for the treatment of head and neck squamous cell carcinoma (HNSCC). Of these, therapeutics targeting the epidermal growth factor receptor (EGFR) have been studied most extensively and some agents have demonstrated measurable clinical effectiveness. However, molecular studies designed to define HNSCC patient subcohorts of likely responders to EGFR-targeted therapy have not identified molecular signatures that correlate with clinical response. Here, the authors summarise the relevant clinical findings and highlight reported molecular correlative studies for EGFR-targeted therapeutics for HNSCC. The authors focus especially on molecular markers evaluated for association with clinical response and include data from EGFR-targeted clinical studies in other cancer sites that they anticipate will be of interest to the head and neck cancer research and treatment communities.

背景与目标： ：几种针对分子的疗法已在临床试验中用于治疗头颈部鳞状细胞癌（HNSCC）。其中，针对表皮生长因子受体（EGFR）的疗法已得到最广泛的研究，某些药物已证明可测量的临床有效性。然而，旨在定义可能对EGFR靶向治疗有反应的HNSCC患者亚组的分子研究尚未发现与临床反应相关的分子标志。在这里，作者总结了相关的临床发现，并着重报道了针对HNSCC的EGFR靶向治疗药物的分子相关研究。作者尤其关注评估与临床反应相关的分子标志物，并包括来自其他癌症部位针对EGFR的临床研究的数据，他们预期这将是头颈癌研究和治疗界感兴趣的数据。

BACKGROUND & AIMS: Interleukin-1 (IL-1) has been implicated in chronic and acute cerebral neuropathologies. IL-1 receptor antagonist (IL-1ra), a naturally occurring protein that binds to IL-1 receptors without inducing signal transduction, blocks several actions of IL-1. IL-1ra acts at the local level and it also circulates in the bloodstream. We now report evidence for a biological function of IL-1ra in the brain as an endogenous neuroprotective molecule. Cerebral expression of IL-1ra mRNA is induced rapidly by focal cerebral ischemia in rats, and inhibition of the action of IL-1ra, by passive immuno-neutralization, markedly enhances ischemic damage. To our knowledge this is the first report of an action of endogenous IL-1ra in the brain. Control of IL-1ra expression or action may therefore provide a useful therapeutic strategy to limit acute neurodegeneration.

背景与目标： 白介素-1（IL-1）已牵涉到慢性和急性脑神经病理学。 IL-1受体拮抗剂（IL-1ra）是一种与IL-1受体结合而不诱导信号转导的天然蛋白质，可阻断IL-1的多种作用。 IL-1ra在局部起作用，并且也在血液中循环。我们现在报告IL-1ra在脑中作为内源性神经保护分子的生物学功能的证据。大鼠局灶性脑缺血可快速诱导IL-1ra mRNA的脑表达，并且被动免疫中和抑制IL-1ra的作用可显着增强缺血性损伤。据我们所知，这是大脑中内源性IL-1ra作用的首次报道。因此，控制IL-1ra的表达或作用可能为限制急性神经变性提供了有用的治疗策略。

BACKGROUND & AIMS: :A structure-activity relationship (SAR) study was performed principally at the N1 position of N1-arylsulfonyl-N2-[1-(1-naphthyl)ethyl]-trans-1,2-diaminocyclohexanes, a new family of calcilytics acting at the calcium sensing receptor (CaSR). The most active compound in this series was the 4-(trifluoromethoxy)benzenesulfonyl derivative 7e, which displayed an IC50 of 5.4 +/- 0.5 microM with respect to the inhibition of calcium-induced tritiated inositol phosphate ([3H]IP) accumulation in Chinese hamster ovarian (CHO) cells expressing the CaSR. Replacement of the sulfonamide linkage of this compound by a carboxamide led to a 6-fold increase in activity (7m, IC50 = 0.9 +/- 0.2 microM). Among the carboxamides synthesized, one of the most active compounds was the 4-chlorophenylcarboxamide (1S,2S,1'R)-7n (Calhex 231, IC50 = 0.33 +/- 0.02 microM). The absolute configuration of (1S,2S,1'R)-7n was deduced from an X-ray crystallographic study of one of the diastereomers of compound 7d. The stereochemical preference for the (1S,2S,1'R)-isomers can be rationalized on the basis of a three-dimensional model of the calcilytic binding pocket of the CaSR. Removal of the C-1' methyl group or replacement of the 1-naphthyl group by a 2-naphthyl or biphenyl moiety led to appreciable loss of calcilytic activity. Compounds 7e, 7m, and Calhex 231 did not stimulate [3H]IP accumulation in CHO cells expressing or not expressing the CaSR.

背景与目标： ：结构-活性关系（SAR）研究主要在N1-芳基磺酰基-N2- [1-（1-（萘基）乙基]-反式-1,2-二氨基环己烷的N1位置进行，钙敏感受体（CaSR）。该系列中活性最高的化合物是4-（三氟甲氧基）苯磺酰基衍生物7e，在抑制中国仓鼠中钙诱导的tri化肌醇磷酸酯（[3H] IP）积累方面，其IC50为5.4 /-0.5 microM。表达CaSR的卵巢（CHO）细胞。该化合物的磺酰胺键被羧酰胺取代导致活性增加了6倍（7m，IC50 = 0.9 /-0.2 microM）。在合成的羧酰胺中，活性最高的化合物之一是4-氯苯基羧酰胺（1S，2S，1'R）-7n（Calhex 231，IC50 = 0.33 /-0.02 microM）。 （1S，2S，1'R）-7n的绝对构型是由化合物7d的一种非对映异构体的X射线晶体学研究得出的。 （1S，2S，1'R）异构体的立体化学偏好可以根据CaSR钙解结合口袋的三维模型来合理化。除去C-1'甲基或用2-萘基或联苯部分取代1-萘基导致明显的钙解活性损失。化合物7e，7m和Calhex 231不会刺激表达或不表达CaSR的CHO细胞中的[3H] IP积累。

BACKGROUND & AIMS: :Granulocyte colony-stimulating factor (GCSF) administration has been linked to the development of monosomy 7 in severe congenital neutropenia and aplastic anemia. We assessed the effect of pharmacologic doses of GCSF on monosomy 7 cells to determine whether this chromosomal abnormality developed de novo or arose as a result of favored expansion of a preexisting clone. Fluorescence in situ hybridization (FISH) of chromosome 7 was used to identify small populations of aneuploid cells. When bone marrow mononuclear cells from patients with monosomy 7 were cultured with 400 ng/ml GCSF, all samples showed significant increases in the proportion of monosomy 7 cells. In contrast, bone marrow from karyotypically normal aplastic anemia, myelodysplastic syndrome, or healthy individuals did not show an increase in monosomy 7 cells in culture. In bone marrow CD34 cells of patients with myelodysplastic syndrome and monosomy 7, GCSF receptor (GCSFR) protein was increased. Although no mutation was found in genomic GCSFR DNA, CD34 cells showed increased expression of the GCSFR class IV mRNA isoform, which is defective in signaling cellular differentiation. GCSFR signal transduction via the Jak/Stat system was abnormal in monosomy 7 CD34 cells, with increased phosphorylated signal transducer and activation of transcription protein, STAT1-P, and increased STAT5-P relative to STAT3-P. Our results suggest that pharmacologic doses of GCSF increase the proportion of preexisting monosomy 7 cells. The abnormal response of monosomy 7 cells to GCSF would be explained by the expansion of undifferentiated monosomy 7 clones expressing the class IV GCSFR, which is defective in signaling cell maturation.

背景与目标： ：粒细胞集落刺激因子（GCSF）的管理与严重先天性中性粒细胞减少和再生障碍性贫血的7号单体病的发展有关。我们评估了药理学剂量的GCSF对7号单体细胞的影响，以确定这种染色体异常是从头发展还是由于已有克隆的有利扩增而出现。 7号染色体的荧光原位杂交（FISH）用于鉴定少量非整倍体细胞。当用400 ng / ml GCSF培养来自第7号单体患者的骨髓单个核细胞时，所有样品均显示出第7号单体细胞的比例显着增加。相反，来自核型正常再生障碍性贫血，骨髓增生异常综合症或健康个体的骨髓在培养物中未显示单核7细胞的增加。在骨髓增生异常综合症和7号单体症患者的骨髓CD34细胞中，GCSF受体（GCSFR）蛋白增加。尽管在基因组GCSFR DNA中未发现突变，但CD34细胞显示GCSFR IV类mRNA亚型的表达增加，这在信号细胞分化中是有缺陷的。通过Jak / Stat系统进行的GCSFR信号转导在7号单体CD34细胞中异常，磷酸化信号转导子的增加和转录蛋白STAT1-P的激活以及相对于STAT3-P的STAT5-P的增加。我们的结果表明，GCSF的药理剂量增加了先前存在的单核7细胞的比例。 7号单体细胞对GCSF的异常应答可以通过表达IV类GCSFR的未分化的7号单体克隆的扩增来解释，该克隆在信号细胞的成熟中是有缺陷的。

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: BACKGROUND AND OBJECTIVES:We tested the hypothesis that single nucleotide polymorphisms (SNPs) in the calcium-sensing receptor (CASR) alter the response to the calcimimetic cinacalcet. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS:We analyzed DNA samples in the Evaluation of Cinacalcet HCl Therapy to Lower Cardiovascular Events (EVOLVE) trial, a randomized trial comparing cinacalcet to placebo on a background of usual care. Of the 3883 patients randomized, 1919 (49%) consented to DNA collection, and samples from 1852 participants were genotyped for 18 CASR polymorphisms. The European ancestry (EA; n=1067) and African ancestry (AfAn; n=405) groups were assessed separately. SNPs in CASR were tested for their association with biochemical measures of mineral metabolism at baseline, percent change from baseline to 20 weeks, and risk of clinical fracture as dependent variables. RESULTS:There were modest associations of CASR SNPs with increased baseline serum parathyroid hormone and bone alkaline phosphatase primarily with the minor allele in the EA group (all P≤0.03), but not in the AfAn sample. In contrast, there was a modest association of decreased baseline serum calcium and FGF23 with CASR SNPs (P=0.04) primarily with the minor allele in the AfAn but not in the EA sample. The minor allele of two SNPs was associated with decreased percent reduction in parathyroid hormone from baseline to 20 weeks in the EA population (P<0.04) and this was not altered with cinacalcet. In both EA and AfAn, the same SNP (rs9740) was associated with decreased calcium with cinacalcet treatment (EA and AfAn P≤0.03). Three SNPs in high linkage disequilibrium were associated with a higher risk of clinical fracture that was attenuated by cinacalcet treatment in the EA sample (P<0.04). CONCLUSIONS:These modest associations, if validated, may provide explanations for differences in CKD-mineral bone disorder observed in EA and AfAn populations, and for differential biochemical responses to calcimimetics.

背景与目标： 背景与目的：我们检验了钙敏感受体（CASR）中的单核苷酸多态性（SNP）改变对拟钙剂西那卡塞的反应的假说。
设计，地点，参与者和测量：我们在Cinacalcet HCl降低心血管事件治疗评估（EVOLVE）试验中分析了DNA样品，该试验是在常规护理的背景下将cinacalcet与安慰剂进行比较的随机试验。在3883名随机分组的患者中，有1919名（49％）同意收集DNA，并对来自1852名参与者的样本进行了18种CASR多态性的基因分型。欧洲血统（EA； n = 1067）和非洲血统（AfAn； n = 405）分别进行了评估。测试了CASR中的SNP与基线时矿物质代谢的生化指标，从基线到20周的百分比变化以及作为相关变量的临床骨折风险之间的相关性。
结果：在EA组中，CASR单核苷酸多态性与基线血清甲状旁腺激素和骨碱性磷酸酶升高之间存在适度的相关性，主要与次要等位基因相关（均P≤0.03），而在AfAn样品中则无此关联。相反，基线血清钙和FGF23降低与CASR SNPs（P = 0.04）之间存在适度的联系，主要与AfAn中的次要等位基因有关，而与EA样品中无关。在EA人群中，两个SNP的次要等位基因与甲状旁腺激素从基线降低到20周的百分比降低有关（P <0.04），而西那卡塞不变。在EA和AfAn中，相同的SNP（rs9740）与西那卡塞治疗引起的钙减少有关（EA和AfAnP≤0.03）。高连锁不平衡中的三个SNP与EA样品中的西那卡塞特治疗减弱的临床骨折风险较高相关（P <0.04）。
结论：这些适度的关联性如果得到验证，则可以解释EA和AfAn人群中CKD矿物骨骼疾病的差异，以及对拟钙剂的不同生化反应。

BACKGROUND & AIMS: :Given the paucity of data on the distribution of serotonin (5-HT) receptors of type 6 (5-HT(6)) in the human brain, the aim of this study was to investigate their distribution in postmortem human prefrontal cortex, striatum and hippocampus by either immunohistochemical or immunofluorescence techniques. The brain samples were obtained from 6 subjects who had died for causes not involving primarily or secondarily the CNS. The 5-HT(6) receptor distribution was explored by the [(125)I]SB-258585 binding to brain membranes followed by immunohistochemical and immunofluorescence evaluations. A specific [(125)I]SB-258585 binding was detected in all the regions under investigation, whilst the content in the hippocampus and cortex being about 10-30 times lower than in the striatum. Immunohistochemistry and double-label immunofluorescence microscopy experiments, carried out in the prefrontal cortex and hippocampus only, since data in the striatum were already published, showed the presence of 5-HT(6) receptors in both pyramidal and glial cells of prefrontal cortex, while positive cells were mainly pyramidal neurons in the hippocampus. The heterogeneous distribution of 5-HT(6) receptors provides a preliminary explanation of how they might regulate different functions in different brain areas, such as, perhaps, brain trophism in the cortex and neuronal firing in the hippocampus. This study, taking into account all the limitations due to the postmortem model used, represents the starting point to explore the 5-HT(6) receptor functionality and its sub-cellular distribution.

背景与目标： ：鉴于缺乏6型血清素（5-HT）受体（5-HT（6））在人脑中的分布的数据，本研究的目的是研究它们在死后人类前额叶皮层，纹状体中的分布免疫组织化学或免疫荧光技术检测海马和海马。脑样本是从6名因主要或次要不涉及CNS的原因死亡的受试者中获得的。通过[（125）I] SB-258585与脑膜的结合，然后进行免疫组织化学和免疫荧光评估，探索了5-HT（6）受体的分布。在所有研究区域中均检测到特定的[（125）I] SB-258585结合，而海马和皮质中的含量比纹状体低约10-30倍。仅在前额叶皮层和海马中进行的免疫组织化学和双标记免疫荧光显微镜实验，因为纹状体中的数据已经发表，显示在前额叶皮层的锥体细胞和神经胶质细胞中都存在5-HT（6）受体，而阳性细胞主要是海马中的锥体神经元。 5-HT（6）受体的异质分布为它们如何在不同的大脑区域调节不同的功能提供了初步的解释，例如，大脑皮层的营养和海马神经元的放电。这项研究，考虑到由于使用死后模型的所有限制，代表了探索5-HT（6）受体功能及其亚细胞分布的起点。

BACKGROUND & AIMS: :There is an abundance of evidence showing that repeated use of 3,4-methlylenedioxymethamphetamine (MDMA; ecstasy) is associated with brain dysfunction, memory disturbance, locomotor hyperactivity, and hyperthermia. MDMA is toxic to both the serotonergic neurons and dopaminergic system. Adenosine is an endogenous purine nucleoside with a neuromodulatory function in the central nervous system. Nuclear factor kappa-B (NF-kB) plays a pivotal role in the initiation and perpetuation of an immune response by triggering the expression of major inflammatory mediators such as cytokines, chemokines, and adhesion molecules. Here, we investigated the effects of the A2a adenosine receptor (A2a-R) agonist (CGS) and antagonist (SCH) on NF-kB expression after MDMA administration. Male Sprague-Dawley rats were injected to MDMA (10 mg/kg) followed by intraperitoneal injection of either CGS or SCH (0.03 mg/kg each) to animals. The hippocampi were then removed for western blot and RT- PCR analyses. MDMA significantly elevated NF-kB expression. Our results show that administration of CGS following MDMA significantly elevated the NF-kB expression both at mRNA and protein levels. By contrast, administration of the A2a-R antagonist SCH resulted in a decrease in the NF-kB levels. Taken together, these results indicate that, co-administration of A2a agonist (CGS) can protect against MDMA neurotoxic effects by increasing NF-kB expression levels; suggesting a potential application for protection against the neurotoxic effects observed in MDMA users.

背景与目标： ：有大量证据表明，反复使用3,4-亚甲基二氧基甲基苯丙胺（MDMA;摇头丸）与脑功能障碍，记忆障碍，运动过度活跃和体温过高有关。 MDMA对血清素能神经元和多巴胺能系统均具有毒性。腺苷是在中枢神经系统中具有神经调节功能的内源性嘌呤核苷。核因子κB（NF-kB）通过触发主要炎症介质（例如细胞因子，趋化因子和粘附分子）的表达，在免疫反应的启动和持久中起关键作用。在这里，我们研究了MDMA给药后A2a腺苷受体（A2a-R）激动剂（CGS）和拮抗剂（SCH）对NF-kB表达的影响。将雄性Sprague-Dawley大鼠注射至MDMA（10 mg / kg），然后向动物腹膜内注射CGS或SCH（每只0.03 mg / kg）。然后取出海马，进行western blot和RT-PCR分析。 MDMA显着提高了NF-kB的表达。我们的结果表明，在MDMA之后施用CGS可以显着提高mRNA和蛋白水平上的NF-kB表达。相反，施用A2a-R拮抗剂SCH导致NF-kB水平降低。综上所述，这些结果表明，共同使用A2a激动剂（CGS）可以通过增加NF-kB表达水平来预防MDMA神经毒性作用。提示了针对MDMA用户观察到的抗神经毒性作用的潜在应用。

BACKGROUND & AIMS: :In human cerebral cortex slices noradrenaline, isoproterenol (a beta-adrenergic agonist), dopamine, apomorphine (a dopaminergic agonist), and serotonin stimulated cyclic AMP formation: noradrenaline greater than or equal to isoproterenol greater than dopamine = apomorphine = serotonin. Clonidine (and alpha-adrenergic agonist) was ineffective in stimulating cyclic AMP formation in temporal cortex slices. The stimulatory effect of noradrenaline and isoproterenol was blocked by propranolol (a beta-adrenergic blocker) but not by phentolamine (an alpha-adrenergic blocker). Pimozide (a selective dopaminergic antagonist) inhibited the increase of cyclic AMP formation induced by dopamine or apomorphine but not that induced by noradrenaline, isoproterenol, or serotonin. Neither propranolol or phentolamine had any effect on dopamine- or serotonin-stimulated cyclic AMP formation. Chlorpromazine blocked the increase of cyclic AMP formation induced by noradrenaline, dopamine or serotonin, while cyproheptadine, a putative central serotonergic antagonist, was ineffective. These observations suggest that there may be at least two monoamine-sensitive adenylate cyclases in human cerebral cortex which have the characteristics of a beta-adrenergic and a dopaminergic receptor, respectively, and also possibly a serotonergic receptor.

背景与目标： 在人大脑皮层切片中，去甲肾上腺素，异丙肾上腺素（β-肾上腺素能激动剂），多巴胺，阿扑吗啡（多巴胺能激动剂）和5-羟色胺刺激的环状AMP形成：去甲肾上腺素大于或等于异丙肾上腺素大于多巴胺=阿扑吗啡= 5-羟色胺。可乐定（和α-肾上腺素激动剂）在刺激颞叶皮质切片中的环状AMP形成方面无效。去甲肾上腺素和异丙肾上腺素的刺激作用被普萘洛尔（一种β-肾上腺素受体阻滞剂）阻断，但未被酚妥拉明（一种α-肾上腺素能阻断剂）阻断。 Pimozide（一种选择性的多巴胺能拮抗剂）抑制多巴胺或阿扑吗啡诱导的环状AMP形成的增加，但不抑制去甲肾上腺素，异丙肾上腺素或5-羟色胺诱导的环状AMP形成的增加。普萘洛尔或酚妥拉明均未对多巴胺或5-羟色胺刺激的环状AMP的形成产生任何影响。氯丙嗪阻断了去甲肾上腺素，多巴胺或5-羟色胺诱导的环AMP形成的增加，而环丙庚定（一种假定的中枢5-羟色胺能拮抗剂）无效。这些观察结果表明，人大脑皮层中可能至少存在两种​​单胺敏感的腺苷酸环化酶，它们分别具有β-肾上腺素能和多巴胺能受体的特征，还可能具有血清素能受体的特征。

BACKGROUND & AIMS: BACKGROUND AND OBJECTIVES:It is well documented that the decrease of glucocorticoid receptor (GR) can lead to Yin-yang Deficiency Syndrome in the Traditional Chinese Medicine theory. Youguiyin a famous and traditional Chinese medicine is often used to treat Yang Deficiency Syndrome, but its mechanism of action and target remains unknown. We aimed to establish one cell model whose the GR gene had been decreased and observe the effect of the traditional Chinese medicine Youguiyin on GR gene expression at mRNA level. MATERIALS AND METHODS:Established recombinant plasmids of GR gene by combination use RNAi and the Cre-LoxP system, stably transfected the recombinant plasmids and Cre-ERT2 plasmid into the murine macrophage RAW264.7 cells and selected with G418 and hygromycin B respectively, then used 4-TH to induce Cre-ERT2 plasmid to express. RT-PCR and Western blot methods to validate the change of GR gene at mRNA and protein level were employed. Feed orally SD male rats with Youguiyin and got their blood serum, used these blood serum to culture RAW264.7-Cre-GR (-,F) cells, detected the change of GR gene at mRNA level by RT-PCR. RESULTS:We successfully constructed two recombinant plasmids of GR gene which can make GR gene's expression to decrease significantly in RAW264.7 cells. The blood serum which contained Youguiyin can enhance the expression of GR mRNA. CONCLUSIONS:Combination use RNAi and the Cre-LoxP system can decrease GR gene's expression in the murine macrophage RAW264.7 cells. Youguiyin can enhance the expression of GR mRNA.

背景与目标： 背景与目的：已有大量文献报道，糖皮质激素受体（GR）的降低可导致中医理论的阴阳虚证。右归饮是一种著名的传统中药，常被用于治疗阳虚证，但其作用机理和靶向作用尚不清楚。我们的目的是建立一个GR基因减少的细胞模型，并观察中药油桂银在mRNA水平上对GR基因表达的影响。
材料与方法：利用RNAi和Cre-LoxP系统联合建立GR基因重组质粒，将重组质粒和Cre-ERT2质粒稳定转染到鼠巨噬细胞RAW264.7细胞中，分别用G418和潮霉素B筛选，然后使用4-TH诱导Cre-ERT2质粒表达。采用RT-PCR和Western blot方法验证GR基因在mRNA和蛋白质水平的变化。用尤桂银口服SD雄性大鼠，取血清，用这些血清培养RAW264.7-Cre-GR（-，F）细胞，通过RT-PCR检测GR基因在mRNA水平的变化。
结果：成功构建了两个GR基因重组质粒，可以使GR基因在RAW264.7细胞中的表达显着降低。含有幽归饮的血清可以增强GR mRNA的表达。
结论：RNAi和Cre-LoxP系统联合使用可降低GR基因在小鼠巨噬细胞RAW264.7细胞中的表达。右归饮可以增强GR mRNA的表达。

BACKGROUND & AIMS: :Sensitizing mutations in the epidermal growth factor receptor (EGFR) predict response to EGFR tyrosine kinase inhibitors (TKIs) and both first- and second-generation TKIs are available as first-line treatment options in patients with advanced EGFR-mutant non-small cell lung cancer. Eventual resistance develops with multiple mechanisms identifiable both upon repeat biopsy and in plasma circulating tumor DNA. The T790M gatekeeper mutation is responsible for almost 60% of cases. A number of third-generation TKIs are in clinical development, and osimertinib has been approved by the US Food and Drug Administration for the treatment of patients with EGFR T790M mutant lung cancer after failure of initial EGFR kinase therapy. Resistance mechanisms are being identified to these novel agents, and the treatment landscape of EGFR-mutant lung cancer continues to evolve. The sequence of EGFR TKIs may change in the future and combination therapies targeting resistance appear highly promising.

背景与目标： ：表皮生长因子受体（EGFR）中的致敏突变预测对EGFR酪氨酸激酶抑制剂（TKIs）的反应，对于晚期EGFR突变的非小细胞患者，第一代和第二代TKI均可作为一线治疗选择肺癌。最终的耐药性会通过重复活检和血浆循环肿瘤DNA鉴定出多种机制发展。 T790M Gatekeeper突变负责将近60％的病例。许多第三代TKI正在临床开发中，奥西替尼已被美国食品和药物管理局批准用于治疗初始EGFR激酶治疗失败后的EGFR T790M突变型肺癌患者。人们已经确定了对这些新型药物的耐药机制，EGFR突变型肺癌的治疗前景也在不断发展。 EGFR TKIs的序列可能会在未来发生变化，针对耐药性的联合疗法似乎很有希望。