A microaliquoting technique for precise histological annotation and optimization of cell content in frozen tissue specimens

Knowledge of the exact cell content of frozen tissue samples is of growing importance in genomic research. We developed a microaliquoting technique to measure and optimize the cell composition of frozen tumor specimens for molecular studies. Frozen samples of 31 mesothelioma cases were cut in alternating thin and thick sections. Thin sections were stained and evaluated visually. Thick sections, i.e., microaliquots, were annotated using bordering stained sections. A range of cellular heterogeneity was observed among and within samples. Precise annotation of samples was obtained by integration and compared to conventional single face and “front and back” section estimates of cell content. Front and back estimates were more highly correlated with block annotation by microaliquoting than were single face estimates. Both methods yielded discrepant estimates, however, and for some studies may not adequately account for the heterogeneity of mesothelioma or other malignancies with variable cellular composition. High yield and quality RNA was extracted from precision annotated, tumor-enriched subsamples prepared by combining individual microaliquots with the highest tumor cellularity estimates. Microaliquoting provides accurate cell content annotation and permits genomic analysis of enriched subpopulations of cells without fixation or amplification.     

Is the edTPA the Right Choice for Evaluating Teacher Readiness?

The purpose of this article is to describe and analyze the edTPA, a performance assessment created by the Stanford Center for Assessment, Learning, and Equity (SCALE) and administered by Pearson, Inc., to assess the professional readiness of student teachers. We challenge claims made in support of using this assessment, specifically within the context of arts teacher preparation programs, and we address areas of immediate critical concern to make alternative recommendations.     

Preparation of optically active 4-chlorophenylalanine from its racemate by deracemization technique using transformant Escherichia coli cells

We have developed a method for the preparation of l-4-chlorophenylalanine from its racemate with Escherichia coli cells expressing a single foreign gene. l-4-Chlorophenylalanine was obtained in a high optical yield by the inversion of configuration of its d-form via the tandem reactions catalyzed by d-amino acid dehydrogenase (DadA) and branched-chain amino acid aminotransferase (BCAAT). While we constructed a plasmid for BCAAT utilizing the gene from Sinorhizobium meliloti ATCC 51124, the first enzyme DadA was the dadA-gene product from E. coli host cell itself, which was activated by the addition of l-alanine in the growth medium.     

牙体预备对高嵌体修复下颌第一磨牙力学影响的初步研究

目的：通过有限元法,探讨高嵌体修复根管治疗后的下颌第一磨牙后,牙体预备形式对牙体组织受力的影响,为临床提供力学理论依据。方法：模拟右侧下颌第一磨牙的三维有限元模型,磨牙存在合面I类洞缺损,根管治疗后采用高嵌体修复,分别设计三种牙体预备形式,即覆盖全部牙尖、覆盖全部功能尖和覆盖部分功能尖的高嵌体修复,对模型进行加载,观察牙体组织的应力大小及分布。结果：覆盖全部牙尖高嵌体修复时牙釉质出现一个应力集中区,其他两种设计出现两个应力集中区。垂直载荷下,釉质的最大主应力在全部牙尖组分别低于其他两组92．29％和89．76％;舌颊向载荷下比其他两组分别降低80．91％和76．53％。三组牙本质的应力集中区趋于一致。垂直载荷下,牙本质的最大主应力在全部牙尖组分别高出其他两组12．92％和14．73％;舌颊向载荷下较其他两组高1．26％和5．08％。结论：从生物力学角度,覆盖全部牙尖的高嵌体更有利于牙体硬组织的受力,可以更好的对牙釉质起到保护作用。     

七鳃鳗Lja-SHP2分子鉴定、重组表达及免疫学研究

高等脊椎动物的蛋白酪氨酸磷酸酶SHP2(SH2 domain-containing protein-tyrosine phosphatase-2)由ptpn11基因编码,催化酪氨酸残基去磷酸化,与其他能催化酪氨酸磷酸化的蛋白酪氨酸激酶共同调节机体内多种信号通路的信号传导。以往研究表明,SHP2在高等脊椎动物T细胞和B细胞的激活与信号转导过程中起着重要作用。为了研究无颌类脊椎动物日本七鳃鳗(Lampetra japonica)中与SHP2同源的分子——Lja-SHP2在免疫应答反应中的作用,本研究通过PCR扩增获取其Lja-SHP2开放阅读框序列,并构建到原核表达载体pET-32a中,成功在大肠杆菌中实现重组蛋白表达并制备了其兔源多克隆抗体。用混合菌免疫刺激日本七鳃鳗后,通过实时荧光定量PCR和免疫印迹方法检测了Lja-SHP2在日本七鳃鳗免疫相关组织中mRNA和蛋白水平表达谱。结果显示,混合菌免疫刺激后,Lja-SHP2 mRNA和蛋白表达在外周血白细胞和髓样小体中无显著变化,而在鳃组织中显著性上调(P<0.05),说明Lja-SHP2在混合菌刺激后主要参与了鳃组织的免疫应答反应。为了进一步探究Lja-SHP2与淋巴细胞亚群免疫应答反应的相关性,本研究分别使用B细胞有丝分裂原脂多糖(lipopolysaccharide,LPS)和T细胞的有丝分裂原植物凝集素(phytohemagglutinin,PHA)免疫刺激日本七鳃鳗。经LPS免疫刺激后,与对照组相比,白细胞中Lja-SHP2蛋白表达显著上调,鳃组织和髓样小体没有显著性差异表达;但经PHA免疫刺激后,与对照组相比,白细胞、鳃组织和髓样小体3种组织中Lja-SHP2均有上调,尤其在白细胞中上调最为显著,大约是对照组的2.5倍,说明Lja-SHP2参与了日本七鳃鳗由PHA介导的免疫应答反应。由于PHA能刺激日本七鳃鳗鳃组织中VLRA+淋巴细胞的活化,这表明Lja-SHP2可能参与了PHA介导的VLRA+淋巴细胞亚群的免疫应答反应。上述研究结果为进一步探索Lja-SHP2在七鳃鳗免疫应答过程中的功能奠定了基础,也为揭示SHP2分子家族的系统发生及探索高等脊椎动物适应性免疫系统的早期发生及其进化历程提供一定的线索。     

3-Acyl-5-hydroxybenzofuran derivatives as potential anti-estrogen breast cancer agents: A combined experimental and theoretical investigation

We first report the application of 3-acyl-5-hydroxybenzofurans as a scaffold to develop potential drugs for breast cancer. Seven novel derivative compounds were synthesized by using a microwave-assisted synthesis method. Those compounds exhibited different antiproliferation against human breast cancer MCF-7 cells, with the best activity of IC₅₀ = 43.08 μM for compound 1. A Quantum Mechanics Polarized Ligand Docking (QPLD) study was carried out to investigate the binding interactions between these compounds and estrogen receptor alpha (ERα). The simulation results showed that the trend of receptor–ligand binding interactions was same as that of their antiproliferative activities. A detailed analysis indicated that compound 1 possesses the highest Van der Waals and hydrogen bond interactions compared to the other six compounds and better inhibitors are achievable by enhancing the hydrogen bond interactions. Based on these results, we addressed that 3-acyl-5-hydroxybenzofuran is an attractive scaffold for designing drugs against breast cancer.     

Determination of the binding mode and interacting amino-acids for dibasic H3 receptor antagonists

Due to its involvement in major CNS functions, the histamine H3 receptor (H3R) is the subject of intensive medicinal chemistry investigation, supported by the range of modern drug discovery tools, such as receptor modeling and ligand docking. Although the receptor models described to date share a majority of common traits, they display discrete alternatives in amino-acid conformation, rendering ligand binding modes quite different. Such variations impede structure-based drug design in the H3R field. In the present study, we used a combination of medicinal chemistry, receptor-guided and ligand-based methods to elucidate the binding mode of antagonists. The approaches converged towards a ligand orientation perpendicular to the membrane plane, bridging Glu206 of the transmembrane helix 5 to acidic amino acids of the extracellular loops. This consensus will help future structure-based drug design for H3R ligands.     

小金海棠果实表皮制片方法的比较研究

为了简便而快速制作适用于光学显微镜观测的苹果属植物果实表皮标本,采用7种方法分别对小金海棠果实表皮进行整体制片后于光学显微镜下观察其形态特征,比较制片效果。结果表明:用离析刮片法、指甲油印迹法、刮片法和煮沸剥离刮片法制备的标本在光学显微镜下均呈现出清晰结构,其中煮沸剥离刮片法的制片效果最佳。因此,煮沸剥离刮片法最适用于苹果属植物果实表皮整体制片,该方法操作简便且效率高。     

北虫草复合制剂对小鼠免疫功能的影响

探讨北虫草复合制剂对小鼠免疫功能的调节作用。建立小鼠免疫力降低的动物模型,实验分6组:对照组、衰老/免疫抑制模型组、白介素-2(IL-2)和北虫草复合制剂的不同剂量组。采用称重法测定免疫器官重量,计算胸腺指数和脾指数。小鼠溶血素抗体生成采用绵羊红细胞致敏法。北虫草复合制剂对小鼠脾脏指数和胸腺指数的影响:实验组(50.2±2.4与27.6±3.6)明显高于模型组(45.6±4.8与23.6±3.6),单位:(mg/10 g体重),差异有显著性(P<0.05)。对小鼠溶血素抗体生成的影响:实验组(53.53±7.8)高于药物对照组(36.50±7.3)。北虫草复合制剂能恢复衰老小鼠的胸腺指数和脾脏指数,增强免疫抑制小鼠血清溶血素含量,因此,对免疫功能低下的小鼠模型具有免疫调节作用。