The Importance of GLUT3 for De Novo Lipogenesis in Hypoxia-Induced Lipid Loading of Human Macrophages

Atherosclerotic lesions are characterized by lipid-loaded macrophages (foam cells) and hypoxic regions. Although it is well established that foam cells are produced by uptake of cholesterol from oxidized LDL, we previously showed that hypoxia also promotes foam cell formation even in the absence of exogenous lipids. The hypoxia-induced lipid accumulation results from increased triglyceride biosynthesis but the exact mechanism is unknown. Our aim was to investigate the importance of glucose in promoting hypoxia-induced de novo lipid synthesis in human macrophages. In the absence of exogenous lipids, extracellular glucose promoted the accumulation of Oil Red O-stained lipid droplets in human monocyte-derived macrophages in a concentration-dependent manner. Lipid droplet accumulation was higher in macrophages exposed to hypoxia at all assessed concentrations of glucose. Importantly, triglyceride synthesis from glucose was increased in hypoxic macrophages. GLUT3 was highly expressed in macrophage-rich and hypoxic regions of human carotid atherosclerotic plaques and in macrophages isolated from these plaques. In human monocyte-derived macrophages, hypoxia increased expression of both GLUT3 mRNA and protein, and knockdown of GLUT3 with siRNA significantly reduced both glucose uptake and lipid droplet accumulation. In conclusion, we have shown that hypoxia-induced increases in glucose uptake through GLUT3 are important for lipid synthesis in macrophages, and may contribute to foam cell formation in hypoxic regions of atherosclerotic lesions.     

High-throughput molecular identification of fish eggs using multiplex suspension bead arrays

The location and abundance of fish eggs provide information concerning the timing and location of spawning activities and can provide fishery-independent estimates of spawning biomass. However, the full value of egg and larval surveys is severely restricted because many species' eggs and larvae are morphologically similar, making species-level identification difficult. Recent efforts have shown that nearly all species of fish may be identified by mitochondrial DNA (mtDNA) sequences (e.g. via 'DNA barcoding'). By taking advantage of a DNA barcode database, we have developed oligonucleotide probes for 23 marine fish species that produce pelagic eggs commonly found in California waters. Probes were coupled to fluorescent microspheres to create a suspension bead array. Biotin-labelled primers were used to amplify the mitochondrial cytochrome oxidase subunit I (COI) and 16S ribosomal rRNA genes from individual fish eggs. The amplicons were then hybridized to the bead array, and after the addition of a reporter fluorophore, samples were analysed by flow cytometry with Luminex 100 instrumentation. Probes specifically targeted eggs that are abundant and/or from morphologically indistinguishable species pairs. Results showed that the 33 different probes designed for this study accurately identified all samples when PCR was successful. Suspension bead arrays have a number of benefits over other methods of molecular identification; these arrays permit high multiplexing, simple addition of new probes, high throughput and lower cost than DNA sequencing. The increasing availability of DNA barcode data for numerous fish faunas worldwide suggests that bead arrays could be developed and widely used for fish egg, larval and tissue identifications.     

鹟科五种鸟线粒体DNA序列变化与亲缘关系的研究

首次对鹟科（Muscicapidae）画眉亚科三种鸟----画眉（Garrulax canorus）、红嘴相思鸟（Leiothrix lutea）、棕头雅雀(Paradoxornis webbiana);鸫亚科的乌鸫（Turdinae merula）线粒体DNA的12S rRNA基因片段的DNA序列进行了测定,并与北美画眉亚科弯嘴鹛属（Pomatostomus）的灰冠弯嘴鹛（P.temporalis）同源序列进行分析比较并构建分子进化树。实验结果与传统形态学论述存在一定的差异,与同工酶研究结果相同：画眉与红嘴相思鸟的亲缘关系最近,棕头鸦雀较乌鸫与画眉亚科的关系更近,为将其置于画眉亚科提供了分子证据。 Abstract：The classfication on Muscicapinae disturbed us for a long time.The important thing is to find an appropriate character to resolve it.It is the first time that we testified the mitochondrial 12S rRNA gene fragment sequence of four birds (G.canorus,L.lutea,P.webbianus and T.merula-owing to Muscicapinae) and aligned with the same sequence of P.temporalis.Using the mitochondrial 12S rRNA sequences date,we built phylogenetic trees.The results indicated that that the relationship between Garrulax canorus and Leiothrix lutea in the four birds was closer which is congruent with morphology.According to the results of isoenzyme and DNA sequencing data,we can draw a conclusion that Paradoxornis webbianus has closer relationship to Timaliinae than that of Turdus merula,and that putting Paradoxonithinae to Timaliinae was reasonable.     

罗非鱼TRAF3基因的表达及功能研究

肿瘤坏死因子受体(TNFR)相关因子3(TRAF3)是多种免疫途径中的关键调控因子。从尼罗罗非鱼(Oreochromis niloticus)中克隆获得了TRAF3基因, 命名为OnTRAF3(GeneBank No. MN258118), 该基因包含1个环指结构域、1个锌指结构域、1个卷曲螺旋和MATH结构域。多序列比对表明, OnTRAF3与其他已知的TRAF3蛋白具有高度的相似性, 尤其是MATH结构域。实时荧光定量PCR(qRT-PCR)分析显示, OnTRAF3在各组织中广泛分布, 且在脑、皮肤、肠和鳃中表达量较高。在无乳链球菌诱导后, 多个组织中的OnTRAF3表达量出现了不同程度的上调, 说明OnTRAF3参与了罗非鱼的抗菌免疫应答。亚细胞定位实验显示, OnTRAF3分布在HEK293的细胞质和细胞核中。此外, 荧光素酶报告基因实验结果显示, 野生型(WT)OnTRAF3可显著激活NF-κB信号, 而coiled-coil和MATH 结构域缺失后, 依然能够显著激活NF-κB 活性, 而RING 和Zinc缺失后, 该激活作用则明显减弱, 表明RING 和Zinc 结构域是OnTRAF3在免疫信号通路中行使功能的关键结构域。研究为探索TRAF3在罗非鱼免疫应答中的功能提供了重要的基础。     

哺乳动物的核移植

哺乳动物的核移植陆长富卢光（湖南医科大学人类生殖工程研究室,长沙４１００７８）ＮｕｃｌｅａｒＴｒａｎｓｐｌａｎｔａｔｉｏｎｉｎＭａｍｍａｌｉａｎＬＵＣｈａｎｇｆｕＬＵＧｕａｎｇｘｉｕ（ＨｕｍａｎＲｅｐｒｏｄｕｃｔｉｖｅＥｎｇｉｎｅｒｉｎｇＬａｂｏｒ...     

Genome structure of the genus Azospirillum

Azospirillum species are plant-associated diazotrophs of the alpha subclass of Proteobacteria. The genomes of five of the six Azospirillum species were analyzed by pulsed-field gel electrophoresis. All strains possessed several megareplicons, some probably linear, and 16S ribosomal DNA hybridization indicated multiple chromosomes in genomes ranging in size from 4.8 to 9.7 Mbp. The nifHDK operon was identified in the largest replicon.