Dermal carotenoid measurements via pressure mediated reflection spectroscopy

We describe a reflection‐based method for the quantitative detection of carotenoid antioxidants in living human skin. The skin tissue site of interest is illuminated with broad‐band white light spanning the spectral range from 350–850 nm and the spectral composition of the diffusively reflected light is analyzed in real time. Topical pressure is applied to temporarily squeeze blood out of the illuminated tissue volume. In this way the influence of oxy‐hemoglobin on the reflection spectra is effectively reduced. After a short optical clearing time the carotenoid absorption becomes easily discernable in a 460–500 nm spectral window and its optical density can be calculated with high accuracy. Our empirical methodology provides a non‐invasive rapid determination of skin carotenoid levels, can be used to monitor skin carotenoid concentration changes over time in response to carotenoid containing natural or supplemental diets, and is easily adaptable for applications in clinical and field settings. (© 2012 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

内皮细胞特异性启动子pvWF和pVE的克隆及表达

目的:克隆并验证内皮细胞(Endothelial Cells,ECs)特异性启动子,为转染人胚胎干细胞(h ESC)后实时监测ECs的定向分化情况以及利用干细胞实施血友病A的基因治疗研究提供基础。方法:通过酶消化法原代分离人脐静脉内皮细胞(HUVECs),结合RT-PCR和免疫荧光验证分离后的HUVECs表达内皮细胞特异性标志基因血管性血友病因子(v WF)和血管内皮钙粘素(VE-cadherin/CDH5)。抽提HUVECs的g DNA,通过PCR扩增内皮细胞特异性表达基因v WF和VE-cadherin转录起始位点上游不同大小的启动子片段,将其取代报告基因载体p EGFP-N1中的广谱启动子CMV,构建4个质粒,即pv WF-1、pv WF-2、p VE-1、p VE-2,分别转染HUVECs和h ESCs,48 h后观察并比较各启动子片段启动绿色荧光蛋白GFP表达情况,筛选最具特异性及转录活性的启动子片段。结果:通过酶消化法,本研究成功分离出具有典型上皮样细胞的HUVECs。RT-PCR和免疫荧光结果表明HUVECs特异性表达v WF和VE-cadherin。酶切及测序证实所构建的4个含ECs特异性启动子片段的质粒与理论序列相符,通过核转染至HUVECs及h ESCs后,48 h后观察到所克隆的VE-cadherin 2105bp启动子片段具有内皮细胞表达的特异性和较强的转录活性。结论:本研究成功筛选出具有内皮细胞表达特异性及较强转录活性的启动子片段。     

CHARGE CONTRAST IMAGING OF EXCEPTIONALLY-PRESERVED FOSSILS

Abstract:  Charge contrast images are a variant of secondary electron images acquired by operating a variable pressure scanning electron microscope in low vacuum mode; i.e. a gas is present in the specimen chamber. Spatial variation in the amount of charge that accumulates on the surface of the specimen is expressed as differences in greyscale tone; areas that are charging less are darker in tone. The precise mechanisms by which charge contrast images are generated are not known fully. Various different properties of a mineral may create a charge contrast; electrical conductivity is known to be one potentially important variable. As carbon is highly conductive but typical host lithologies (carbonates, silicates) less so or dielectric, the technique is potentially very suitable for imaging organically preserved fossils such as those from the Solite and Jehol biotas. It can also be applied to Burgess Shale fossils: complex films comprising 'aluminosilicates' with or without carbonaceous remains. Charge contrast images reveal anatomical detail not visible using either optical or other scanning electron microscope-based imaging methods.     

抗生素单剂与联合治疗社区获得性肺炎的临床疗效观察

目的:观察β内酰胺类联合大环内酯类与β内酰胺类单剂治疗社区获得性肺炎的临床疗效。方法:收集我院呼吸内科住院的社区获得性肺炎患者262例,随机分为单剂治疗组和联合治疗组。两组患者均给予抗感染、对症支持治疗,其中单剂治疗组抗生素采用β内酰胺酶类单剂治疗,联合治疗组采用β内酰胺酶类和大环内酯类联合治疗。所有患者统计年龄、性别、基础疾病、病情严重程度,并记录治疗前与治疗后不同住院天数的血常规、C反应蛋白的变化,统计住院天数。结果:联合治疗患者一般基本情况(年龄、性别、基础疾病、病情严重程度)与单剂治疗组相比,P0.05,差异均无统计学意义。治疗前及治疗后首日C反应蛋白和白细胞计数二者相比,P0.05,差异均无统计学意义;联合治疗组与单剂治疗组相比,C反应蛋白在住院第2、4、6、9天,白细胞计数在住院第2、4、6天,均明显下降,住院天数缩短,P0.05,差异均有统计学意义。对于中、重度CAP(CRB65≥2),联合治疗组与单剂治疗组相比,治疗后C反应蛋白以及白细胞计数明显减少,P均0.05,差异有统计学意义;对于轻度CAP(CRB65=0~1),两组治疗后CRB和白细胞计数相比,P均0.05,差异无统计学意义。结论:β内酰胺类联合大环内酯类治疗治疗社区获得性肺炎临床疗效优于β内酰胺类单剂治疗。     

Enhancement of YD spin relaxation by the CaMn4 cluster in photosystem II detected at room temperature: A new probe for the S-cycle

The long-lived, light-induced radical Y_D^• of the Tyr161 residue in the D2 protein of Photosystem II (PSII) is known to magnetically interact with the CaMn₄ cluster, situated ∼ 30 Å away. In this study we report a transient step-change increase in Y_D^• EPR intensity upon the application of a single laser flash to S₁ state-synchronised PSII-enriched membranes from spinach. This transient effect was observed at room temperature and high applied microwave power (100 mW) in samples containing PpBQ, as well as those containing DCMU. The subsequent decay lifetimes were found to differ depending on the additive used. We propose that this flash-induced signal increase was caused by enhanced spin relaxation of Y_D^• by the OEC in the S₂ state, as a consequence of the single laser flash turnover. The post-flash decay reflected S₂ → S₁ back-turnover, as confirmed by their correlations with independent measurements of S₂ multiline EPR signal and flash-induced variable fluorescence decay kinetics under corresponding experimental conditions. This flash-induced effect opens up the possibility to study the kinetic behaviour of S-state transitions at room temperature using Y_D^• as a probe.     

Scanning Electron Microscope Study of the Root-knot Nematode (Meloidogyne incognita) on Tomato Root

This study examines the types of structural information that can be gained by utilizing the scanning electron microscope (SEM) and a cryofracture technique to examine the host-parasite interaction. Roots of tomato, Lycopersicon esculentum cv. Marglobe, were cultured aseptically and inoculated with the root-knot nematode, Meloidogyne incognita. Twenty-four hours to four weeks after inoculation, developing galls were removed from the cultures and processed for SEM observation. The cryofracture technique was used to reveal internal structural features within the developing galls. The results illustrate structural details concerning penetration of the roots, differentiation of syncytia, and development of the nematodes beginning with the second-stage larvae and ending with adult egg-laying females.     

Effect of Electroconvulsive Shock on Mitochondrial Respiratory Chain in Rat Brain

It is well described that impairment of energy production has been implicated in the pathogenesis of a number of diseases. Although several advances have occurred over the past 20 years concerning the use and administration of electroconvulsive therapy (ECT) to minimize its side effects, little progress has been made in understanding its mechanism of action. In this work, our aim was to measure the activities of mitochondrial respiratory chain complexes II and IV and succinate dehydrogenase from rat brain after acute and chronic electroconvulsive shock (ECS). Our results showed that mitochondrial respiratory chain enzymes activities were increased after acute ECS in hippocampus, striatum and cortex of rats. Besides, we also demonstrated that complex II activity was increased after chronic ECS in cortex, while hippocampus and striatum were not affected. Succinate dehydrogenase, however, was inhibited after chronic ECS in striatum, activated in cortex and not affected in hippocampus. Finally, complex IV was not affected by chronic ECS in hippocampus, striatum and cortex. Our findings demonstrated that brain metabolism is altered by ECS.     

A temperature‐dependent conformational change of NADH oxidase from Thermus thermophilus HB8

Using molecular dynamics simulations and steady‐state fluorescence spectroscopy, we have identified a conformational change in the active site of a thermophilic flavoenzyme, NADH oxidase from Thermus thermophilus HB8 (NOX). The enzyme's far‐UV circular dichroism spectrum, intrinsic tryptophan fluorescence, and apparent molecular weight measured by dynamic light scattering varied little between 25 and 75°C. However, the fluorescence of the tightly bound FAD cofactor increased approximately fourfold over this temperature range. This effect appears not to be due to aggregation, unfolding, cofactor dissociation, or changes in quaternary structure. We therefore attribute the change in flavin fluorescence to a temperature‐dependent conformational change involving the NOX active site. Molecular dynamics simulations and the effects of mutating aromatic residues near the flavin suggest that the change in fluorescence results from a decrease in quenching by electron transfer from tyrosine 137 to the flavin. Proteins 2012. © 2011 Wiley Periodicals, Inc.     

直接重整细胞核程序的诱导性多能干细胞研究进展

诱导性多能干细胞（induced pluripotent stem cells, iPS）是分化细胞在外源性因子作用下,经直接细胞核程序重整而重新获得分化潜能的干细胞,具有很重要的应用前景。介绍了iPS诱导方法从转录因子、RNA结合蛋白、小分子化合物、到信号传导通路的发展过程,以及在提高生物安全性方面的改进。iPS的生成在细胞学上表现为渐进的、时间依赖的过程,同细胞的分化状态密切相关;然而,iPS同胚胎干细胞表遗传特征并非完全相同。iPS的进展结合基因治疗和细胞治疗的成果已应用到动物疾病模型的治疗。