聚甲基丙烯酰胺包覆的ZnO发光量子点及其在细胞成像中的应用

通过溶胶-凝胶法合成了在水溶液中稳定发光、荧光颜色可调的ZnO@polymer核壳型纳米粒子, 并研究了这种量子点对人类宫颈癌HeLa细胞的毒性以及细胞吞噬后激光共聚焦成像的效果. 这种荧光探针的外壳是一种通过配位键与ZnO内核结合的共聚物, 该共聚物外层是亲水的聚甲基丙烯酰胺, 内层是疏水的聚甲基丙烯酸酯. 细胞毒性实验证明, 该材料有良好的生物兼容性, 适用于生物荧光标记. 共聚焦荧光成像结果显示, 这些纳米粒子可以穿透HeLa细胞膜并在细胞质中稳定发光.     

基于原子力显微镜的人外周血CD8+T细胞形貌观察与力谱分析

体外分离人外周血CD8+T细胞,用植物凝结素(PHA)刺激活化,利用原子力显微镜(AFM)观察CD8+T细胞的形貌,并结合针尖修饰技术对力谱进行分析,确定了CD8抗原分子的分布.结果表明,与静息的CD8+T细胞相比,活化后的CD8+T细胞直径和高度增大,细胞表面变得更粗糙;CD8抗原-抗体的相互作用力大约是非特异性黏附力的5倍,活化后的CD8+T细胞上特异性黏附力(即CD8抗原分子位置)分布不均匀,其表面的CD8抗原分子分布以纳米团簇分布为主,CD8分子聚集更为明显,CD8+T细胞上CD8抗原-抗体相互作用力不随着活化而发生明显变化,说明CD8抗原-抗体之间具有高选择性.原子力显微镜为特异性T细胞的抗原识别和活化研究提供了一种新手段,能够使T细胞抗原识别和活化的机制得到更好地阐明.     

键合紫杉醇的纳米胶束与C6胶质瘤细胞的相互作用

制备了键合紫杉醇(PTX)的聚乙二醇-聚乳酸嵌段共聚物(PEG-PLA/PTX)的纳米胶束, 采用四氮唑(MTT)比色法、流式细胞术、透射电镜及激光共聚焦显微镜等技术, 考察了PEG-PLA/PTX胶束对C6胶质瘤细胞的影响, 包括C6细胞超微结构的变化和细胞周期的改变, 以及纳米颗粒在细胞内的分布, 探讨了PEG-PLA/PTX胶束对肿瘤细胞的作用机理. 结果表明, PEG-PLA/PTX胶束进入到C6细胞内, 聚集于细胞浆中, 通过与细胞核中DNA的作用改变细胞生长的周期, 造成在G2-M期的阻滞, 引起细胞的凋亡. 因此, PEG-PLA/PTX胶束有望用于脑胶质瘤的化疗.     

Determination of Apoptotic Mechanism of Action of Tetrabromobisphenol A and Tetrabromobisphenol S in Human Peripheral Blood Mononuclear Cells: A Comparative Study

Background: Tetrabromobisphenol A (TBBPA) is the most commonly used brominated flame retardant (BFR) in the industry. TBBPA has been determined in environmental samples, food, tap water, dust as well as outdoor and indoor air and in the human body. Studies have also shown the toxic potential of this substance. In search of a better and less toxic BFR, tetrabromobisphenol S (TBBPS) has been developed in order to replace TBBPA in the industry. There is a lack of data on the toxic effects of TBBPS, while no study has explored apoptotic mechanism of action of TBBPA and TBBPS in human leukocytes. Methods: The cells were separated from leucocyte-platelet buffy coat and were incubated with studied compounds in concentrations ranging from 0.01 to 50 µg/mL for 24 h. In order to explore the apoptotic mechanism of action of tested BFRs, phosphatidylserine externalization at cellular membrane (the number of apoptotic cells), cytosolic calcium ion and transmembrane mitochondrial potential levels, caspase-8, -9 and -3 activation, as well as PARP-1 cleavage, DNA fragmentation and chromatin condensation in PBMCs were determined. Results: TBBPA and TBBPS triggered apoptosis in human PBMCs as they changed all tested parameters in the incubated cells. It was also observed that the mitochondrial pathway was mainly involved in the apoptotic action of studied compounds. Conclusions: It was found that TBBPS, and more strongly TBBPA, triggered apoptosis in human PBMCs. Generally, the mitochondrial pathway was involved in the apoptotic action of tested compounds; nevertheless, TBBPS more strongly than TBBPA caused intrinsic pathway activation.     

Spectrally and Time-Resolved Fluorescence Imaging of 22-NBD-Cholesterol in Human Peripheral Blood Mononuclear Cells in Chronic Kidney Disease Patients

The interaction of the fluorescent probe 22-NBD-cholesterol with membranes of human peripheral blood mononuclear cells (PBMC) was tested by time- and spectrally resolved fluorescence imaging to monitor the disturbance of lipid metabolism in chronic kidney disease (CKD) and its treatment with statins. Blood samples from healthy volunteers (HV) and CKD patients, either treated or untreated with statins, were compared. Spectral imaging was done using confocal microscopy at 16 spectral channels in response to 458 nm excitation. Time-resolved imaging was achieved by time-correlated single photon counting (TCSPC) following excitation at 475 nm. The fluorescence of 22-NBD-cholesterol was mostly integrated into plasmatic membrane and/or intracellular membrane but was missing from the nuclear region. The presence of two distinct spectral forms of 22-NBD-cholesterol was uncovered, with significant variations between studied groups. In addition, two fluorescence lifetime components were unmasked, changing in CKD patients treated with statins. The gathered results indicate that 22-NBD-cholesterol may serve as a tool to study changes in the lipid metabolism of patients with CKD to monitor the effect of statin treatment.     

毛细管气相色谱法测定健康人外周血单个核细胞磷脂脂肪酸组成

采用毛细管气相色谱法对输注脂肪乳剂的健康志愿者外周血单个核细胞磷脂脂肪酸组成进行了分析测定 ,并将定量结果与输注脂肪乳剂前进行了比较 ,结果表明 :与输注脂肪乳剂前相比 ,连续 7d外周静脉输注 2 0 %脂肪乳剂 (2 5 0mL d) ,外周血单个核细胞数无明显变化 ;外周血单个核细胞磷脂酰乙醇胺中棕榈酸(P <0 .0 5 )和油酸 (P <0 .0 1 )明显增加 ,硬脂酸下降 ;磷脂酰胆碱中棕榈油酸 (P <0 .0 5 )和亚麻酸 (P <0 .0 5 )明显增加 ,而两种磷脂中花生四烯酸及其它多不饱和脂肪酸、饱和脂肪酸与不饱和脂肪酸的比值以及脂肪酸的不饱和指数均未发生明显变化。     

A Novel Method for the Detection of Chlorpyrifos by Combining Quantum Dot-labeled Molecularly Imprinted Polymer with Flow Cytometry

Uniform-sized fluorescent molecularly imprinted polymers were prepared by one-step swelling and suspension polymerization, while chlorpyrifos, methacrylic acid, ethylene glycol dimethacrylate, and oil-soluble CdSe/ZnS quantum dots were used as the carrier, template molecule, functional monomer, cross-linker, and fluorophor, respectively. The morphology, adsorption dynamics, binding ability, and selectivity of quantum dot-labeled molecularly imprinted polymers were evaluated. The dosage of quantum dots for labeling the molecularly imprinted polymers was optimized. The results showed that the optimized dose of quantum dots was 200?µL using a concentration of 8.0?µM. The microsphere size was approximately 10?µm with a honeycombed surface. The quantum dot-labeled molecularly imprinted polymers had an even brightness and a high selectivity. In the presence of different concentrations of chlorpyrifos, a decrease in the fluorescence intensity of the quantum dot-labeled molecularly imprinted polymer was clearly identified by flow cytometry. The whole detection process was accomplished within 2?h including pretreatment. This method was used for the determination of chlorpyrifos in tap water samples.     

热水提取与超声提取的麦冬多糖结构与构象特征的比较研究

利用热水与超声提取麦冬粗多糖,DEAE-cellulose52纤维素与Sephacryl S-300凝胶柱层析对其进行分离纯化,得到热水提取的麦冬多糖均一组分WPOJ-DS与超声提取的麦冬多糖均一组分UPOJ-DS。采用高效液相色谱法、气相色谱法、红外光谱法、部分酸水解、甲基化分析及核磁共振对WPOJ-DS与UPOJDS的结构进行表征;利用刚果红实验、圆二色谱实验及原子力显微镜对其溶液构象进行了比较研究。结果表明,超声提取对麦冬多糖的分子量、单糖组成摩尔比及构型会产生影响,WPOJ-DS与UPOJ-DS的主链均主要由→6)-D-Glcp(1→和→3,6)-D-Glcf(1→组成,但WPOJ-DS中存在→6)-β-D-Glcp(1→,而UPOJDS中没有;刚果红实验、圆二色谱实验与原子力显微镜结果显示UPOJ-DS存在螺旋结构,而WPOJ-DS没有;且在不同溶液环境中,WPOJ-DS与UPOJ-DS的溶液构象不同。     

Study of the effect of atorvastatin on the interaction between ICAM-1 and CD11b by live-cell single-molecule force spectroscopy

The interaction between the cell adhesion molecule CD11b and its ligand ICAM-1 plays an important role in inflammatory responses in the disease of atherosclerosis. Atorvastatin is a commonly prescribed statin drug which has been considered as one of the most potent therapeutic agents for atherosclerosis due to its lipid-lowering effect. Recently, there is a growing body of evidence that atorvastatin has anti-inflammatory effect. We have applied the advanced method of live-cell single-molecule force spectros...     

Investigation of the Effect of Thermal Annealing on Poly(3‐hexylthiophene) Nanofibers by Scanning Probe Microscopy: From Single‐Chain Conformation and Assembly Behavior to the Interfacial Interactions with Graphene Oxide

Poly(3‐hexylthiophene) (P3HT) has been widely used in devices owing to its excellent properties and structural features. However, devices based on pure P3HT have not exhibited high performance. Strategies, such as thermal annealing and surface doping, have been used to improve the electrical properties of P3HT. In this work, different from previous studies, the effect of thermal annealing on P3HT nanofibers are examined, ranging from the single polymer chain conformation to chain packing, and the interfacial interactions with graphene oxide (GO) at nanoscale dimensions, by using scanning tunneling microscopy (STM), atomic force microscopy (AFM) and Kelvin probe force microscopy (KPFM). High‐resolution STM images directly show the conformational changes of single polymer chains after thermal annealing. The morphology of P3HT nanofibers and the surface potential changes of the P3HT nanofibers and GO is further investigated by AFM and KPFM at the nanoscale, which demonstrate that the surface potentials of P3HT decrease, whereas that of GO increases after thermal annealing. All of the results demonstrate the stronger interfacial interactions between P3HT and GO occur after thermal treatments due to the changes in P3HT chain conformation and packing order.     

A Comparative Study of Functionalized High‐Purity Carbon Nanotubes towards the V(IV)/V(V) Redox Reaction Using Cyclic Voltammetry and Scanning Electrochemical Microscopy

Functionalized high purity carbon nanotubes (CNTs) with various amounts of oxygen containing surface groups were investigated towards the relevant redox reactions of the all‐vanadium redox flow battery. The quinone/hydroquinone redox peaks between 0.0 and 0.7 V vs. Ag|AgCl|KCl_sat. were used to quantifying the degree of functionalization and correlated to XPS results. Cyclic voltammetry in vanadyl sulfate‐containing 3 M H₂SO₄ as a common supporting electrolyte showed no influence of the amount of surface groups on the V(IV)/V(V) redox system. In contrast, the reactions occurring at the negative electrode (V(II)/V(III) and V(III)/V(IV)) are strongly affected by oxygen surface groups. However, under modified experimental conditions, SECM experiments detecting the consumption of VO²⁺ molecules by CNT thin films in pH=2 solution show improved onset potentials with increased surface oxygen content up to ∼ 3 at%. Further increase in surface oxygen up to 8 at% led to minor improvement. These dissimilar results under different experimental conditions are rationalized by suggesting that oxygen functional groups do not form the active site for the V(IV)/V(V) reaction but wetting of the catalyst layer is of high importance.     

Dependence of the Nanoscale Composite Morphology of Fe3O4 Nanoparticle-Infused Lysozyme Amyloid Fibrils on Timing of Infusion: A Combined SAXS and AFM Study

Understanding the formation process and the spatial distribution of nanoparticle (NP) clusters on amyloid fibrils is an essential step for the development of NP-based methods to inhibit aggregation of amyloidal proteins or reverse the assembling trend of the proto-fibrillary complexes that prompts pathogenesis of neuro degeneration. For this, a detailed structural determination of the diverse hybrid assemblies that are forming is needed, which can be achieved by advanced X-ray scattering techniques. Using a combined solution small angle X-ray scattering (SAXS) and atomic force microscopy (AFM) approach, this study investigates the intrinsic trends of the interaction between lysozyme amyloid fibrils (LAFs) and Fe₃O₄ NPs before and after fibrillization at nanometer resolution. AFM images reveal that the number of NP clusters interacting with the lysozyme fibers does not increase significantly with NP volume concentration, suggesting a saturation in NP aggregation on the fibrillary surface. The data indicate that the number of non-adsorbed Fe₃O₄ NPs is highly dependent on the timing of NP infusion within the synthesis process. SAXS data yield access to the spatial distribution, aggregation manner and density of NP clusters on the fibrillary surfaces. Employing modern data analysis approaches, the shape and internal structural morphology of the so formed nanocomposites are revealed. The combined experimental approach suggests that while Fe₃O₄ NPs infusion does not prevent the fibril-formation, the variation of NP concentration and size at different stages of the fibrillization process can impose a pronounced impact on the superficial and internal structural morphologies of these nanocomposites. These findings may be applicable in devising advanced therapeutic treatments for neurodegenerative diseases and designing novel bio-inorganic magnetic devices. Our results further demonstrate that modern X-ray methods give access to the structure of—and insight into the formation process of—biological–inorganic hybrid structures in solution.