Europium complexes as novel indicators of paracellular diffusion

Measurement of paracellular permeation is an important assay for tight-junction investigations of drug toxicity, especially for metal-based drugs, and routine validation of the integrity of cell monolayers for models of drug absorption. Great efforts have been made in discovery and validation of novel paracellular diffusion indicators. In the present work, we prepared three Eu complexes, i.e., [Eu(dtpa)] (dtpa=diethylenetriaminepentaacetic acid), [Eu(dtpa)(BSA)], and [Eu(dtpa)(PLL)] (PLL=poly(L-lysine)), and tested their permeation properties on Madin?Darby canine kidney (MDCK) cells. The experimental results showed that all three probes were nontoxic to MDCK cells, permeated across MDCK monolayer exclusively via the paracellular pathways, and responded well to the changes on tight junction with high correlation of P(app) values to the decrease of trans-epithelial electric resistance (TEER). In addition, time-resolved fluorescence assays were conducted in a high-sensitivity and background-free mode. All these results confirmed the Eu complexes as novel and practical paracellular indicators.     

Endophytic fungal community in stems and leaves of plants from desert areas in China

Endophytic fungi are known to play important ecological roles in protecting plants from various abiotic and biotic stresses. Therefore, it is valuable to investigate the endophytic fungal community associated with plants distributed in harsh environments, such as deserts. Fungal communities in the stems and leaves of ten plant samples belonging to eight species were collected from a desert area in China and tested after plant surface sterilization. The fungal compositions were different among plants. Salsola collina, Suaeda salsa, and Coriospermum declinatum possessed the highest fungal richness. The colonization rates of these samples were high, exceeding 50% in eight of the samples. However, the fungal diversity of the samples was low when measured using Shannon??s index, Fisher??s ??, and Simpson??s index. Alternaria alternata, A. franseriae, Fusarium solani, and a second Fusarium species were most frequently isolated from all samples. The diversity of isolated species was low in desert areas, although the colonization rate was relatively high. It was concluded that fungal communities associated with plants in deserts had low diversity, but a small number of species colonized various plants with a high colonization rate. The Jaccard, Sorensen, and Bray?CCurtis similarity indices for the fungal communities were low between stems and leaves. This indicated that different fungal communities colonized these two tissues. Phoma pomorum and Phoma sp. showed tissue preferences.     

Bisubstrate analog inhibitors of 6-hydroxymethyl-7,8-dihydropterin pyrophosphokinase: new lead exhibits a distinct binding mode

6-Hydroxymethyl-7,8-dihydropterin pyrophosphokinase (HPPK), a key enzyme in the folate biosynthesis pathway catalyzing the pyrophosphoryl transfer from ATP to 6-hydroxymethyl-7,8-dihydropterin, is an attractive target for developing novel antimicrobial agents. Previously, we studied the mechanism of HPPK action, synthesized bisubstrate analog inhibitors by linking 6-hydroxymethylpterin to adenosine through phosphate groups, and developed a new generation of bisubstrate inhibitors by replacing the phosphate bridge with a piperidine-containing linkage. To further improve linker properties, we have synthesized a new compound, characterized its protein binding/inhibiting properties, and determined its structure in complex with HPPK. Surprisingly, this inhibitor exhibits a new binding mode in that the adenine base is flipped when compared to previously reported structures. Furthermore, the side chain of amino acid residue E77 is involved in protein-inhibitor interaction, forming hydrogen bonds with both 2' and 3' hydroxyl groups of the ribose moiety. Residue E77 is conserved among HPPK sequences, but interacts only indirectly with the bound MgATP via water molecules. Never observed before, the E77-ribose interaction is compatible only with the new inhibitor-binding mode. Therefore, this compound represents a new direction for further development.     

Bisubstrate analogue inhibitors of 6-hydroxymethyl-7,8-dihydropterin pyrophosphokinase: New design with improved properties

6-Hydroxymethyl-7,8-dihydropterin pyrophosphokinase (HPPK), a key enzyme in the folate biosynthetic pathway, catalyzes the pyrophosphoryl transfer from ATP to 6-hydroxymethyl-7,8-dihydropterin. The enzyme is essential for microorganisms, is absent from humans, and is not the target for any existing antibiotics. Therefore, HPPK is an attractive target for developing novel antimicrobial agents. Previously, we characterized the reaction trajectory of HPPK-catalyzed pyrophosphoryl transfer and synthesized a series of bisubstrate analog inhibitors of the enzyme by linking 6-hydroxymethylpterin to adenosine through 2, 3, or 4 phosphate groups. Here, we report a new generation of bisubstrate analog inhibitors. To improve protein binding and linker properties of such inhibitors, we have replaced the pterin moiety with 7,7-dimethyl-7,8-dihydropterin and the phosphate bridge with a piperidine linked thioether. We have synthesized the new inhibitors, measured their K(d) and IC(50) values, determined their crystal structures in complex with HPPK, and established their structure-activity relationship. 6-Carboxylic acid ethyl ester-7,7-dimethyl-7,8-dihydropterin, a novel intermediate that we developed recently for easy derivatization at position 6 of 7,7-dimethyl-7,8-dihydropterin, offers a much high yield for the synthesis of bisubstrate analogs than that of previously established procedure.     

小型猪冠脉支架模型中即刻OCT检查评价支架贴壁不良的安全性和有效性

目的评价中华小型猪冠状动脉支架植入术后即刻行光学相干断层成像扫描（OCT）的安全性和有效性。方法健康中华小型猪22只,经股动脉途径行右冠状动脉西罗莫司药物洗脱支架植入术,术后即刻行OCT检查评价支架贴壁不良情况,同时记录OCT检查时间和相关并发症。结果 OCT共检查25段支架,失败3例（1例指引导丝断裂于支架内,2例发生冠脉痉挛导致阻断球囊送入困难未能成像）,OCT检查并发症包括一过性ST段抬高（5例）和室颤（3例）,死亡1例,平均手术时间49.7±12.9 min,其中OCT检查耗时17.9±4.9 min,OCT共检测522 mm支架,定量测量4514个支架丝,其中66个存在贴壁不良,即刻支架贴壁不良率1.46%。结论利用冠脉内OCT技术可以有效评价支架术后即刻贴壁不良情况,安全性良好。     

赖氨酸特异性组蛋白去甲基化酶1作用机制及其生物学功能的研究进展

与其他化学修饰,如乙酰化、磷酸化、泛素化等相似,组蛋白赖氨酸甲基化是一个可以逆转的组蛋白修饰,是一个动态调节的过程。赖氨酸特异性组蛋白去甲基化酶1（lysine specific demethylase 1,LSD1）是一个黄素腺嘌呤二核苷酸（flavin adenine dinulcleotide,FAD）依赖性胺氧化酶,它能够特异性脱去H3K4和H3K9位点上的单甲基化和二甲基化的甲基基团。LSD1参与调控核受体介导的基因转录,并分别维持染色质的活性和非活性状态,被誉为细胞深处的基因＂开关＂。LSD1的功能失衡可引发多种重要生命现象的改变。主要综述LSD1的结构、作用机制及其在肿瘤发生、胚胎发育、体细胞重编程的调控、细胞分裂和造血等过程中生物学功能的研究新进展。     

雷公藤内酯醇干预肾脏疾病的研究进展

雷公藤的提取物在中国被用于治疗肾小球肾炎已经有30年的历史。雷公藤提取物的主要活性成分雷公藤内酯醇具有免疫抑制和抗炎的特性,然而,毒性作用限制了其临床应用。深入研究雷公藤内酯醇的药理机制,将有助于其临床应用和降低毒副反应。对国内外关于雷公藤内酯醇对肾脏影响的研究成果进行了综述。     

Nogo—p4与NgR结合抑制神经干细胞分化为双极形星形胶质细胞的突起生长

目的观察Nogo—p4是否通过与NgR结合的途径对大鼠脊髓来源神经干细胞分化形成双极形星形胶质细胞突起长度产生抑制。方法取4只出生24h内的Wistar大鼠,悬浮培养法培养大鼠脊髓来源的神经干细胞。把神经干细胞分为A、B、C、D四组,A组加入血清,B组加入血清和Nogo—p4,C组神经干细胞经RNA干扰沉默NgR基因后加入血清分化,D组神经干细胞经RNA干扰沉默NgR基因后加入血清和Nogo—p4。分化第7d,GFAP抗体标记星形胶质细胞,使用Image—ProPlus5．0软件测量双极形星形胶质细胞突起长度。结果神经干细胞分化第7d,四组均可形成双极形星形胶质细胞。B组中双极形星形胶质细胞的突起长度明显短于其它各组。A、C、D组中双极形星形胶质细胞的突起长度没有显著差异。结论Nogo—p4经与NgR结合途径显著抑制脊髓来源神经干细胞分化成的双极形星形胶质细胞的突起生长。     

大连地区感染性腹泻病原监测结果分析

目的 了解大连地区感染性腹泻病原菌的种类和流行特征,为感染性腹泻的预防控制和临床治疗提供依据.方法 采用常规粪便培养方法,对分离菌做血清学分型,用MicroScan WalkAway-40全自动细菌鉴定及药敏分析仪对分离菌做生化鉴定,对检出的志贺菌做药敏试验,并用WHONET 5.4软件对药敏结果进行统计分析.结果 监测872例腹泻患者样本,检出感染性腹泻病原菌109株,阳性率为12.5％,其中志贺菌所占比例最大,其次为沙门菌,副溶血弧菌排第三.志贺菌对碳青霉烯类、氟喹诺酮类和头孢菌素类等敏感,对青霉素类和复方磺胺类耐药.结论 志贺菌是大连地区感染性腹泻的首要病原菌,沙门菌和副溶血弧菌次之,应依据此监测结果做好大连地区腹泻病的防治工作;同时根据耐药性监测结果合理使用抗生素应对志贺菌,减少耐药菌株出现.