BACKGROUND & AIMS: OBJECTIVE:The purpose of this study was to determine the expression of receptor activator of NFkappaB ligand (RANKL) and osteoprotegerin (OPG) associated with bone destruction in periapical cysts and granulomas. STUDY DESIGN:Forty human dental chronic periapical lesions were collected after periapical surgery. The lesions collected were fixed in 10% buffered formalin and histologically processed. At least 2 sections of each specimen were stained with hematoxylin and eosin for microscopic diagnosis. After that, 10 human periapical granulomas and 10 cysts were selected for immunohistochemical analysis for RANKL, OPG, and CD68+. RESULTS:Polymorphonuclear neutrophils, macrophages, endothelial cells, and lymphocytes were stained for RANKL and OPG in both lesions. Epithelial cells were also stained for RANKL and OPG in periapical cysts. Quantitative analysis was conducted and the results were expressed as a ratio of the number of immunostained cells over the total number of cells in the field (n = 100). The ratio of RANKL+/total cells was higher than OPG+/total cells in periapical granulomas (0.553 +/- 0.153 and 0.483 +/- 0.189, respectively; P < .0012; paired t test) and in cysts (0.519 +/- 0.09 and 0.339 +/- 0.117, respectively; P < .0001; paired t test). The ratios of OPG+/total cells (P < .0001; paired t test) and RANKL+/total cells (P < .0322; paired t test) were greater in granulomas than in cysts. However, the ratio RANKL+/OPG+ in granulomas (1.336 +/- 0.723) and cysts (1.404 +/- 0.385) was not significantly different. The ratio of CD68+/total cells was significantly higher in granulomas (0.381 +/- 0.040) than in cysts (0.307 +/- 0.068) (P < .0001; unpaired t test with Welch correction). CONCLUSION:Taking into account the limitations of the experimental approach employed, our findings indicate the presence of RANKL and OPG in cysts and granulomas, strongly suggesting the involvement of these gene products in the development of periapical lesions.

背景与目标： 目的：本研究旨在确定与根尖周囊肿和肉芽肿中的骨破坏有关的NFkappaB配体（RANKL）和骨保护素（OPG）受体激活剂的表达。
研究设计：根尖周手术后收集了40例人类牙齿慢性根尖周病变。将收集的病灶固定在10％的福尔马林缓冲液中，并进行组织学处理。每个标本至少2个切片用苏木精和曙红染色以进行显微镜诊断。之后，选择10个人根尖肉芽肿和10个囊肿进行RANKL，OPG和CD68的免疫组织化学分析。
结果：在两个病变中，多形核中性粒细胞，巨噬细胞，内皮细胞和淋巴细胞均进行了RANKL和OPG染色。还对上皮周囊肿中的上皮细胞进行了RANKL和OPG染色。进行定量分析，结果表示为免疫染色细胞数与现场细胞总数之比（n = 100）。根尖肉芽肿中RANKL /总细胞的比率高于OPG /总细胞的比率（分别为0.553 /-0.153和0.483 /-0.189; P <.0012;成对t检验）和囊肿中（0.519 /-0.09和0.339 / -分别为0.117； P <.0001；成对t检验）。肉芽肿中OPG /总细胞（P <.0001；成对t检验）与RANKL /总细胞（P <.0322；成对t检验）的比率大于囊肿。然而，肉芽肿（1.336 /-0.723）和囊肿（1.404 /-0.385）中的RANKL / OPG比率没有显着差异。肉芽肿中CD68 /总细胞的比率（0.381 /-0.040）显着高于囊肿（0.307 /-0.068）（P <.0001；采用Welch校正的未配对t检验）。
结论：考虑到所用实验方法的局限性，我们的发现表明囊肿和肉芽肿中存在RANKL和OPG，强烈暗示这些基因产物参与了根尖周病变的发展。

BACKGROUND & AIMS: :An immediately inducible gene by gonadotropin was isolated from rat ovaries primed with pregnant mare serum gonadotropin (PMSG) by using a subtraction cloning procedure. Homology analysis revealed that the gene is a rat homologue of scavenger receptor class B-I, which was recently identified as a specific receptor for high density lipoprotein (HDL). The structure of rat SRBI was determined by nucleotide sequence analysis of full-length cDNAs for SRBI. Northern blot analysis revealed that rat SRBI mRNA levels were rapidly and strongly increased within 3 h by the injection of PMSG. In situ hybridization study revealed that SRBI mRNA was strongly induced in theca interna cells of immature rat ovary stimulated with 30 IU of PMSG for 6 h. SRBI mRNA expression was also observed in corpora lutea of the adult rat ovary. These findings indicate that expression of SRBI mRNA is restricted to and induced in the ovarian steroidogenic cell types where cholesterol is used as a substrate for synthesis of steroid hormones. Our data strongly suggest that SRBI may play a significant role in the ovarian steroidogenesis by mediating selective uptake of cholesterol from HDL to ovarian theca interna cells or to corpus luteum.

背景与目标： ：通过减法克隆程序，从用怀孕母马血清促性腺激素（PMSG）引发的大鼠卵巢中分离出由促性腺激素立即诱导的基因。同源性分析显示该基因是清道夫受体B-I类的大鼠同源物，最近被鉴定为高密度脂蛋白（HDL）的特异性受体。通过SRBI全长cDNA的核苷酸序列分析确定大鼠SRBI的结构。 Northern印迹分析表明，通过注射PMSG，大鼠SRBI mRNA水平在3小时内迅速而强烈地增加。原位杂交研究表明，SRBI mRNA在30 IU PMSG刺激6 h的未成熟大鼠卵巢的内膜细胞中被强烈诱导。在成年大鼠卵巢的黄体中也观察到了SRBI mRNA表达。这些发现表明，SRBI mRNA的表达受限于卵巢类固醇生成细胞类型并在其中被诱导，其中胆固醇被用作类固醇激素合成的底物。我们的数据有力地表明，SRBI可能通过介导HDL对胆固醇和卵巢黄体的选择性摄取来介导胆固醇的选择性摄取，从而在卵巢类固醇生成中发挥重要作用。

BACKGROUND & AIMS: :Burkholderia pseudomallei is a facultative intracellular pathogen and the causative agent of melioidosis, a spectrum of potentially fatal diseases endemic in Northern Australia and South-East Asia. We demonstrate that B. pseudomallei rapidly modifies infected macrophage-like cells in a manner analagous to osteoclastogenesis. These alterations include multinucleation and the expression by infected cells of mRNA for factors required for osteoclastogenesis: the chemokines monocyte chemotactic protein 1 (MCP-1), macrophage inflammatory protein 1 gamma (MIP-1gamma), 'regulated on activation normal T cell expressed and secreted' (RANTES) and the transcription factor 'nuclear factor of activated T-cells cytoplasmic 1' (NFATc1). An increase in expression of these factors was also observed after infection with Burkholderia thailandensis. Expression of genes for the osteoclast markers calcitonin receptor (CTR), cathepsin K (CTSK) and tartrate-resistant acid phosphatase (TRAP) was also increased by B. pseudomallei-infected, but not by B. thailandensis-infected cells. The expression by B. pseudomallei-infected cells of these chemokine and osteoclast marker genes was remarkably similar to cells treated with RANKL, a stimulator of osteoclastogenesis. Analysis of dentine resorption by B. pseudomallei-induced osteoclast-like cells revealed that demineralization may occur but that authentic excavation does not take place under the tested conditions. Furthermore, we identified and characterized lfpA (for lactonase family protein A) in B. pseudomallei, which shares significant sequence similarity with the eukaryotic protein 'regucalcin', also known as 'senescence marker protein-30' (SMP-30). LfpA orthologues are widespread in prokaryotes and are well conserved, but are phylogenetically distinct from eukaryotic regucalcin orthologues. We demonstrate that lfpA mRNA expression is dramatically increased in association with macrophage-like cells. Mutation of lfpA significantly reduced expression of the tested host genes, relative to the response to wild-type B. pseudomallei. We also show that lfpA is required for optimal virulence in vivo.

背景与目标： ：伪狂犬病是一种兼职的细胞内病原体，是类鼻oid病的病原体，类li虫病是北澳大利亚和东南亚流行的一系列潜在致命性疾病。我们证明假性芽孢杆菌以与破骨细胞发生类似的方式快速修饰感染的巨噬细胞样细胞。这些改变包括多核化和破骨细胞形成所需因子的感染细胞mRNA表达：趋化因子单核细胞趋化蛋白1（MCP-1），巨噬细胞炎性蛋白1γ（MIP-1gamma），“对激活的正常T细胞的表达和分泌”（RANTES）和转录因子“活化T细胞胞质1的核因子”（NFATc1）。在感染了伯克霍尔德菌之后，这些因子的表达也增加了。破伤风标记的降钙素受体（CTR），组织蛋白酶K（CTSK）和抗酒石酸的酸性磷酸酶（TRAP）的破骨细胞标志物的基因表达也增加了，而被泰国芽孢杆菌（B. thailandensis）感染的细胞却没有。这些趋化因子和破骨细胞标志物基因的经假芽孢杆菌感染的细胞的表达与破骨细胞形成刺激物RANKL处理的细胞非常相似。假性麦芽孢杆菌诱导的破骨细胞样细胞对牙本质的吸收分析表明，可能会发生脱矿质，但在测试条件下不会发生真正的挖掘。此外，我们在假苹果芽孢杆菌中鉴定并鉴定了lfpA（用于内酯酶家族蛋白A），它与真核蛋白“ regucalcin”（也称为“衰老标记蛋白-30”（SMP-30））具有显着的序列相似性。 LfpA直向同源物广泛存在于原核生物中，并且保守性很强，但在系统发育上与真核瑞古钙素直向同源物不同。我们证明，与巨噬细胞样细胞相关联，lfpA mRNA表达显着增加。相对于对野生型假单胞菌的反应，lfpA的突变显着降低了测试宿主基因的表达。我们还表明，lfpA是体内最佳毒力所必需的。

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: :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。