High Precision,Electrochemical Detection of Reversible Binding of Recombinant Proteins on Wide Bandgap GaN Electrodes Functionalized with Biomembrane Models

We report a novel hybrid charge sensor realized by the deposition of phospholipid monolayers on highly doped n‐GaN electrodes. To detect the binding of recombinant proteins with histidine‐tags, lipid vesicles containing chelator lipids were deposited on GaN electrodes pre‐coated with octadecyltrimethoxysilane monolayers. Owing to its optical transparency, GaN allows the confirmation of the fluidity of supported membranes by fluorescence recovery after photo‐bleaching (FRAP). The electrolyte‐(organic) insulator‐semiconductor (EIS) setup enables one to transduce variations in the surface charge density ΔQ into a change in the interface capacitance ΔC _p and, thus, the flat‐band potential ΔU _FB. The obtained results demonstrate that the membrane‐based charge sensor can reach a high sensitivity to detect reversible changes in the surface charge density on the membranes by the formation of chelator complexes, docking of eGFP with histidine tags, and cancellation by EDTA. The achievable resolution of ΔQ ≥ 0.1 μC/cm² is better than that obtained for membrane‐functionalized p‐GaAs, 0.9 μC/cm², and for ITO coated with a polymer supported lipid monolayer, 2.2 μC/cm². Moreover, we examined the potential application of optically active InGaN/GaN quantum dot structures, for the detection of changes in the surface potential from the photoluminescence signals measured at room temperature.     

Effective and Selective Anti‐Cancer Protein Delivery via All‐Functions‐in‐One Nanocarriers Coupled with Visible Light‐Responsive,Reversible Protein Engineering

Efficient intracellular delivery of protein drugs and tumor‐specific activation of protein functions are critical toward anti‐cancer protein therapy. However, an omnipotent protein delivery system that can harmonize the complicated systemic barriers as well as spatiotemporally manipulate protein function is lacking. Herein, an “all‐functions‐in‐one” nanocarrier doped with photosensitizer (PS) is developed and coupled with reactive oxygen species (ROS)‐responsive, reversible protein engineering to realize cancer‐targeted protein delivery, and spatiotemporal manipulation of protein activities using long‐wavelength visible light (635 nm) at low power density (5 mW cm^?2). Particularly, RNase A is caged with H₂O₂‐cleavable phenylboronic acid to form 4‐nitrophenyl 4‐(4,4,5,5‐tetramethyl‐1,3,2‐dioxaborolan‐2‐yl)benzyl carbonate (NBC)‐modified RNase (RNBC), which is encapsulated in acid‐degradable, ketal‐crosslinked PEI (KPEI)‐based nanocomplexes (NCs) coated with PS‐modified hyaluronic acid (HA). Such NCs harmonize the critical processes for protein delivery, wherein HA coating renders NCs with long blood circulation and cancer cell targeting, and KPEI enables endosomal escape as well as acid‐triggered intracellular RNBC release. Tumor‐specific light irradiation generates H₂O₂ to kill cancer cells and restore the protein activity, thus achieving synergistic anti‐cancer efficacy. It is the first time to photomanipulate protein functions by coupling ROS‐cleavable protein caging with PS‐mediated ROS generation, and the “all‐functions‐in‐one” nanocarrier represents a promising example for the programmed anti‐cancer protein delivery.     

基于功率型LED的在体整体荧光光学成像

用发光二极管（LED）替代常用的荧光激发光源汞灯,以实现基于LED的整体荧光光学成像（简称整体成像）。首先通过光谱计算分析,探讨了在不同波段与汞灯激发的荧光量相当时LED需要输出的光通量。以此为依据选择LED,构建了基于功率型LED的整体成像系统,用其动态监测绿色荧光蛋白（GFP）标记的肿瘤在裸鼠体内生长和药物治疗过程。实验表明,功率型LED可在类似于整体成像需要较大光功率的场合中应用。光源替代时,若两光源的光谱在选择的波段内分布不一样,则有必要考虑不同波长的光激发荧光效率差异的影响。     

壬基酚对大鼠F1子代脑基因表达的影响

目的:研究壬基酚通过妊娠期暴露是否会影响F1子代的脑发育。方法:对SD孕鼠染毒NP,从妊娠第7天开始持续到产后第二天,分别提取NP组和对照组F1代2d龄大鼠的脑组织mRNA,表达谱基因芯片检测脑基因的表达。用Q-PCR法验证F1雄性子代大脑泛素特异性蛋白酶8、ATP结合盒转运子和内磺肽蛋白这三个下调基因的表达,然后用Western blotting法检测F1子代大脑泛素特异性蛋白酶8的蛋白表达。结果:ATP结合盒转运子(ATP-binding cassette,ABCA)、内璜肽蛋白(Endosulfine alph,Ensa)、泛素特异性蛋白酶8(Ubiquitin Specific Protease 8,USP8/UBPy)基因和蛋白表达下调。结论:NP可影响神经细胞的正常结构和信号转导功能,并提示可以干扰机体内分泌系统、脂质和能量代谢及一系列正常生理功能。     

对NpHR光敏感蛋白具有刺激作用的光电极研制

光电极是研究光遗传学不可或缺的工具之一。波长为460nm的蓝光可以对ChR2神经蛋白产生刺激,而波长为580nm的黄光可以刺激NpHR神经蛋白,对生物体的反应产生抑制。目前,蓝光波段的光电极具有很高的功率密度,可以满足光遗传学的研究,而黄光波段半导体发光器件的发光效率较低,因此对黄光波段的光电极研究较少。在蓝宝石衬底蓝光光电极研制的基础上,通过激发黄色荧光粉和量子点两种形式获得黄光,并通过沉积SiO2/TiO2分布式布拉格反射镜（DBR）有效滤除了蓝光波段的激发光,最后通过沉积Ag金属反射镜增强黄光信号,制备出荧光粉基和量子点基黄光光电极。与荧光粉基黄光光电极相比,量子点基黄光光电极具有更高的功率密度、更窄的光谱半宽和更薄的厚度。在1～10mA的注入电流下,量子点基黄光光电极的功率密度为4.46～15.37mW/mm2,满足刺激NpHR神经蛋白的要求。     

Size Switchable Nanoclusters Fueled by Extracellular ATP for Promoting Deep Penetration and MRI‐Guided Tumor Photothermal Therapy

Protein‐based theranostic agents (PBTAs) exhibit superior performance in the diagnosis and therapy of cancers. However, the in vivo applications of PBTA are largely limited by undesired accumulation, penetration, or selectivity. Here, an ATP‐supersensitive protein cluster is fabricated for promoting PBTA delivery and enhancing magnetic resonance imaging (MRI)‐guided tumor photothermal therapy. Gd³⁺‐ and CuS‐coloaded small bovine serum albumin nanoparticles (GdCuB) are synthesized as the model protein with a size of 9 nm and are encapsulated into charge switchable polycations (DEP) to form DEP/GdCuB nanoclusters of 120 nm. In blood circulation, DEP/GdCuB significantly extends the half‐lifetime and thereby enhances the tumor accumulation of GdCuB. When the clusters reach the tumor site, the extracellular adenosine triphosphate (ATP) can effectively trigger the release of GdCuB, resulting in tumoral deep penetration as well as the activation of T₁‐weighted MRI (r₁ value switched from 2.8 × 10^?3 to 11.8 × 10^?3 m ^?1 s^?1). Furthermore, this delivery strategy also improves the tumoral photothermal therapy efficacy with the MRI‐guided therapy. The study of ATP‐activated nanoclusters develops a novel strategy for tumor deep penetration and on/off imaging of PBTA by size switchable technology, and reveals the potential for MRI‐guided therapy of cancers.     

Managing the Public Sphere: Journalistic Construction of the Great Globalization Debate

There is little consensus on what constitutes open, deliberative media discourse. We offer a simple, measurable, and comparative model based on 3 aspects of source and issue construction in news accounts: access, recognition, and responsiveness. The model is applied to coverage of 2001–2003 World Economic Forum (WEF) meetings and protests against the organization's role in global economic policies. Both demonstrators and WEF participants were granted news access, but WEF actors were recognized more formally and given greater input in news content, including ownership claims to many activist issue positions. Journalistic deference to the WEF communication agenda limited mutual responsiveness. The journalistic process systematically managed the debate about globalization on terms that favored elites over citizen-activists.     

系统性红斑狼疮患者血液中PRDM1和CD138＋浆细胞表达的研究

目的;探讨系统性红斑狼疮（SiE）患者和正常人之间PRDM1和CD138＋浆细胞的差异。方法：40例SLE患者血液标本,30例健康体检血液标本,抽取RNA、利用逆转录聚合酶链反应（RT—PCR）技术研究PRDM1基因表达;收集外周血单个核细胞,利用流式细胞技术研究CD138＋浆细胞量的差异。结果：SLE患者组血液中的P...     

银屑病患者皮损中角蛋白10、14、17及丝聚蛋白表达与分布

目的：研究银屑病患者皮损中角蛋白10、14、17及丝聚蛋白的分布、表达与皮肤屏障功能受损的相关性。方法以20例银屑病患者皮损及10例正常人皮肤为研究对象,运用免疫组化及westerm blot测定两组角蛋白10、14、17及丝聚蛋白的分布与表达。结果：与正常人皮肤相比,银屑病患者的皮损组织中角蛋白10表达和丝聚蛋白表达下调;角蛋白14及17表达上调,但表达程度及部位不同,其差异均具有统计学意义。结论银屑病患者的皮肤屏障功能有受损,可能是银屑病持续发病机制之一。     

乙酰胆碱酯酶分子的原子力显微成像

本文研究了用原子力显微技术对乙酰胆碱酯酶进行成像的方法。在新鲜裂解的云一表面镀一层金膜,金膜与疏基丙酸（ＭＰＡ）的疏基形成Ｓ－Ａｕ键。使ＭＰＡ的游离羧基再通过碳二亚胺反应与ＡＣｈＥ的碱性氨基酸的氨基形成肽键连接,ＡＣｈＥ因而被固定于支持物表面,这样固定的ＡＣｈＥ可获得清晰成像而不脱落。