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Monomeric RGD peptides show unspecific fluid-phase uptake in cells, whereas multimeric RGD peptides are thought to be internalized by integrin-mediated endocytosis. However, a potential correlation between uptake mechanism and molecular mass has been neglected so far. A dual derivatization of peptide c(RGDw(7Br)K) was performed to investigate this. A fluorescent probe was installed by chemoselective Suzuki–Miyaura cross-coupling of the 7-bromotryptophan and a poly(ethylene glycol) (PEG) linker was attached to the lysine residue. Flow cytometry and live cell imaging confirmed unspecific uptake of the small, non-PEGylated peptide, whereas the PEG5000 peptide conjugate unveiled a selective internalization by M21 cells overexpressing αvβ3 and no uptake in αv-deficient M21L cells.  相似文献   
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DC‐SIGN, an antigen‐uptake receptor in dendritic cells (DCs), has a clear role in the immune response but, conversely, can also facilitate infection by providing entry of pathogens into DCs. The key action in both processes is internalization into acidic endosomes and lysosomes. Molecular probes that bind to DC‐SIGN could thus provide a useful tool to study internalization and constitute potential antagonists against pathogens. So far, only large molecules have been used to directly observe DC‐SIGN‐mediated internalization into DCs by fluorescence visualization. We designed and synthesized an appropriate small glycomimetic probe. Two particular properties of the probe were exploited: activation in a low‐pH environment and an aggregation‐induced spectral shift. Our results indicate that small glycomimetic molecules could compete with antigen/pathogen for binding not only outside but also inside the DC, thus preventing the harmful action of pathogens that are able to intrude into DCs, for example, HIV‐1.  相似文献   
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本研究以绿豆为作用载体,研究了肠炎沙门氏菌(ATCC13076)在绿豆芽的不同发芽阶段中的内化定殖能力及消除绿豆芽中定殖的肠炎沙门氏菌的有效方法。结果表明:在绿豆发芽前期的4个不同时段(0~48 h内):吸胀期(G_1,0 h)、萌动期(G_2,12h)、发芽初期(G_3,24 h)、发芽期(G_4,48 h),分别接种10~2、10~4、10~6和10~8 CFU/mL的肠炎沙门氏菌时,其在绿豆芽中的内化定殖能力不同。接种浓度为10~2CFU/mL左右时,肠炎沙门氏菌在发芽初期(G_3)和发芽期(G_4)的内化能力较强,另外两个阶段比较微弱。接种浓度不低于10~4CFU/mL时,肠炎沙门氏菌在吸胀期(G_1)的内化能力最强。在吸胀期(G_1)接种10~8CFU/mL的菌液时,最高内化量可达2.6×10~8CFU/g。紫外照射消毒处理对内化定殖的肠炎沙门氏菌有明显的去除效果,而次氯酸钠溶液和硝酸银溶液浸泡处理对内化的肠炎沙门氏菌的去除效果并不明显。  相似文献   
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This study aimed to determine the activity of one Mycoplasma bovis nuclease encoded by MBOV_RS02825 and its association with cytotoxicity. The bioinformatics analysis predicted that it encodes a Ca2+-dependent nuclease based on existence of enzymatic sites in a TNASE_3 domain derived from a Staphylococcus aureus thermonuclease (SNc). We cloned and purified the recombinant MbovNase (rMbovNase), and demonstrated its nuclease activity by digesting bovine macrophage linear DNA and RNA, and closed circular plasmid DNA in the presence of 10 mM Ca2+ at 22–65 °C. In addition, this MbovNase was localized in membrane and rMbovNase able to degrade DNA matrix of neutrophil extracellular traps (NETs). When incubated with macrophages, rMbovNase bound to and invaded the cells localizing to both the cytoplasm and nuclei. These cells experienced apoptosis and the viability was significantly reduced. The apoptosis was confirmed by activated expression of phosphorylated NF-κB p65 and Bax, and inhibition of Iκβα and Bcl-2. In contrast, rMbovNaseΔ181–342 without TNASE_3 domain exhibited deficiency in all the biological functions. Furthermore, rMbovNase was also demonstrated to be secreted. In conclusion, it is a first report that MbovNase is an active nuclease, both secretory and membrane protein with ability to degrade NETs and induce apoptosis.  相似文献   
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伴随新理念和新技术的不断涌现,信息技术与课程的整合日渐深入,一种新型的教学模式"翻转课堂"应运而生.时下,翻转课堂成为国内外教育改革的新浪潮,为教与学的进一步发展提供了新的思路.本文着眼于探究在翻转课堂的实施过程中要如何翻转才能达到知识传授这一过程的效率最大化以及吸收内化的深入化.教学环节的翻转是课堂教学形式转变的需要,教师自身翻转是实施的前提和关键,学生的翻转是目的和结果.  相似文献   
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本文以C语言程序设计课程中的递归程序教学为例,论述了基于微课开展翻转课堂教学的教学设计及实施方法。  相似文献   
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Hydroxyapatite is the main inorganic component of biological bone and tooth enamel, and synthetic hydroxyapatite has been widely used as biomaterials. In this study, a facile method has been developed for the fabrication of arginine-functionalized and europium-doped hydroxyapatite nanoparticles (Arg-Eu-HAP). The synthesized nanoparticles characterized by transmission electron microscopy, X-ray diffractometry, Fourier transform infrared, and Zeta potential analyzer. Its biological properties with DNA binding, cell toxicity, cell binding and intracellular distribution were tested by agarose gel electrophoresis assay, flow cytometry, and fluorescence microscope and laser scanning confocal microscope. The synthesized Arg-Eu-HAP could effectively bind DNA without any cytotoxicity and be internalized into the cytoplasm and perinuclear of human lung epithelial cells.  相似文献   
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企业环境成本内部化与应对绿色壁垒的措施   总被引:2,自引:0,他引:2  
绿色壁垒的产生有着深刻的社会、政治、经济原因,在本质上具有二重性,在保护环境的同时,也对贸易形成了障碍。面对绿色壁垒给我国带来的挑战,我们应正视其保护环境、促进社会可持续发展的合理性的一面,通过推行环境成本内部化,纠正环境成本外部性造成的产品价格的扭曲和市场的失灵,实现资源的合理配置,从根本上解决环境与经济的协调问题。最终将引导企业向绿色化发展,冲破绿色壁垒,实现经济效益和环境效益的"双赢"。而面对其不合理的一面,我们也应积极采取各种贸易救济措施,同时合理构筑我国的绿色壁垒,以维护自己的合法权益。  相似文献   
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