硫化铜纳米粒子应用于肿瘤诊断和治疗的研究进展

癌症是当今威胁人类健康的主要疾病之一。近年来提出的近红外光介导的光热治疗,能够对肿瘤组织进行定点清除并且对正常组织具有较低的毒副作用,为肿瘤的治疗提供了新的方法。开发具有良好生物相容性的高效光热偶联剂是发展光热治疗的首要条件。随着纳米技术的飞速发展,一些金属纳米结构由于具有独特的光学特性作为光热偶联剂被广泛应用到肿瘤的光热治疗中。然而,成本高昂、制备过程繁琐以及光热稳定性较差等不足,限制了这些纳米材料的进一步应用。最新报道的新型光热偶联剂半导体硫化铜纳米粒子(copper sulfide nanoparticles,CuS NPs),由于其具有制备工艺简单、成本低廉、突出的光热稳定性和良好的生物相容性等优势,成为了当今纳米医学领域研究的热点。本文主要综述了CuS纳米粒子在肿瘤光热治疗和影像诊断方面的应用研究,并对CuS纳米粒子在生物医学领域应用中存在的问题和未来的研究方向进行了展望。     

An impedimetric vascular endothelial growth factor biosensor-based PAMAM/cysteamine-modified gold electrode for monitoring of tumor growth

In the present work, we have developed a new biosensor based on fourth-generation (G4) PAMAM dendrimers for the analysis of vascular endothelial growth factor (VEGF). First, the PAMAM dendrimers were covalently attached to a cysteamine-modified Au electrode by glutaraldehyde. With the help of the amino groups located on its surface, vascular endothelial growth factor receptor-1 (VEGF-R1) was immobilized via glutaraldehyde cross-linking. VEGF-R1 loading was investigated to identify the optimal VEGF-R1 immobilization conditions for the best sensitivity of the new biosensor. In addition, Kramers-Kronig transforms were also analyzed for immobilization and measurement processes. The biosensor had a linear range of 5 to 125 pg/mL VEGF. The fabricated biosensor had good repeatability and reproducibility. Finally, the results for artificial serum samples measured by the present biosensor showed a good recovery for VEGF detection.     

Effect of the degree of cross-linking on the properties of different CLEAs of penicillin acylase

Cross-linked enzyme aggregates (CLEAs) are novel type biocatalysts well suited to catalyze reactions of organic synthesis. Penicillin acylase is a versatile enzyme that can both hydrolyze and synthesize β-lactam antibiotics. CLEAs and CLEAs covered with polyionic polymers (polyethyleneimine and dextran sulfate at two different enzyme to polymer ratios) were prepared at varying cross-linking agent to enzyme ratio: 0.15 and 0.25. Results are presented on the effect of such variables on immobilization yield, specific activity, stability and performance of penicillin acylase CLEAs in the kinetically controlled synthesis of cephalexin. The cross-linking agent to enzyme ratio had no significant effect on the specific activity of the CLEAs, but affected immobilization yield, stability in ethylene glycol medium and conversion yield and productivity in the synthesis of cephalexin, being always higher at the lower cross-linking agent to enzyme ratio. Best results were obtained with CLEAs at 0.15 glutaraldehyde to enzyme protein ratio: specific activity of hydrolysis and synthesis was 708 and 325 UI/g_CLEA respectively, conversion yield was 87%, specific productivity was 5.4 mmol cephalexin/(g_CLEA·h) and 90% of the enzyme remained active after 170 h at operating conditions.     

Artemisia arborescens L essential oil loaded beads: Preparation and characterization

The purpose of this work was to prepare sodium alginate beads as a device for the controlled release of essential oil for oral administration as an antiviral agent. Different formulations were prepared with sodium alginate as a natural polymer and calcium chloride or glutaraldehyde as a cross-linking agent. Loading capacities of between 86% and 100% were obtained in freshly prepared beads by changing exposure time to the cross-linking agent. Drying of the calcium alginate beads caused only a slight decrease in the loading efficiency. The surface morphology of the different bead formulations were studied using scanning electron microscopy (SEM). Stability studies over a 3-month period showed that glutaraldehyde reacted with some components ofArtemisia arborescens L essential oil, changing its composition. Calcium alginate beads showed an in vitro controlled release of the essential oil for the investigated 24 hours, while the use of glutaraldehyde as a cross-linking agent was found not appropriate because of the interactions with azulene derivatives and the low degree of matrix cross-linkage. Published: August 24, 2007     

Enhancing LSPR Sensitivity of Au Gratings through Graphene Coupling to Au Film

A particular interesting plasmonic system is that of metallic nanostructures interacting with metal films. As the localized surface plasmon resonance (LSPR) behavior of gold nanostructures (Au NPs) on the top of a gold thin film is exquisitely sensitive to the spacer distance of the film-Au NPs, we investigate in the present work the influence of a few-layered graphene spacer on the LSPR behavior of the NPs. The idea is to evidence the role of few-layered graphene as one of the thinnest possible spacer. We first show that the coupling to the Au film induces a strong lowering at around 507 nm and sharpening of the main LSPR of the Au NPs. Moreover, a blue shift in the main LSP resonance of about 13 nm is observed in the presence of a few-layered graphene spacer when compared to the case where gold nanostructures are directly linked to a gold thin film. Numerical simulations suggest that this LSP mode is dipolar and that the hot spots of the electric field are pushed to the top corners of the NPs, which makes it very sensitive to surrounding medium optical index changes and thus appealing for sensing applications. A figure of merit of such a system (gold/graphene/Au NPs) is 2.8, as compared to 2.1 for gold/Au NPs. This represents a 33 % gain in sensitivity and opens-up new sensing strategies.     

Colloidal Synthesis of Plasmonic Metallic Nanoparticles

Solutions of Ag and Au nanoparticles are strongly colored because of localized surface plasmon resonance in the UV/visible spectral region. The optical properties of these nanoparticles may be tuned to suit the needs of the application. This article summarizes our work in recent years on the solution synthesis of nanoparticles with tunable optical properties. The systems of interest include zero-dimensional bimetallic Ag–Au nanoparticles with different structures, one-, two-, and three-dimensional anisotropic monometallic Ag or Au nanoparticles. All of these nanosystems were prepared from colloidal synthesis through simple changes in the synthesis conditions. This is a demonstration of the versatility of colloidal synthesis as a convenient scalable technique for tuning the properties of metallic nanoparticles. Zhang, Tan, and Xie contributed equally to this article     

The Tunable and Well-Controlled Surface Plasmon Resonances of Au Hollow Nanostructures by a Chemical Route

Au plasmonic hollow spherical nanostructures were synthesized by electrochemical reduction (GRR, the Galvanic Replacement Reaction) using Ag nanoparticles as templates. From UV-visible absorption spectroscopy, it was found that the surface plasmon resonance (SPR) of gold hollow spherical nanostructures first showed red shift and then blue shift. However, further addition of gold precursor (HAuCl₄) resulted into a red shift of SPR peak. The morphological changes from Ag nanoparticles to Au hollow nanostructures were assessed by transmission electron microscopy (TEM) and energy dispersive X-ray spectroscopy (EDX)analysis. The Mie Scattering theory based simulations of SPR of Au hollow nanostructures were performed which are in good agreement with the experimental observations. Based on the experimental observations and theoretical calculations, a complete growth mechanism for Au hollow nanostructures is proposed.     

Diazo- and azido-functionalized glutaraldehydes as cross-linking reagents and potential fixatives for electron microscopy

The synthesis of diazo and perfluorophenyl azide (PFPA) functionalized glutaraldehydes 7 and 13a-d as new cross-linking reagents for bioconjugation and potential fixatives for electron microscopy is reported. A key step is the generation of the 1,5-dialdehyde structures by oxidative cleavage of the corresponding cyclopentene epoxide using HIO4 in aqueous tetrahydrofuran. A model reaction between 3-substituted glutaraldehyde 14 and 6-aminohexanoic acid resulted in the formation of pyridinium ion containing products with UV spectra comparable to those observed with glutaraldehyde itself. Thus modification of glutaraldehyde in the 3-position most probably did not significantly change its reactivity with amines under chemical-fixation conditions. Fixation of red blood cells by 7 demonstrates that as a fixative, 7 is comparable to glutaraldehyde.     

Tailoring LSPR-Based Absorption and Scattering Efficiencies of Semiconductor-Coated Au nanoshells

Absorption and scattering efficiencies of semiconductor-coated Au nanoshell have been studied by the extended Mie theory for their possible solar cell, optical imaging, and photothermal applications, etc. The effect of Au shell layer thickness, core size, and surrounding medium on the absorption and scattering efficiencies at the localized surface plasmon resonance (LSPR) wavelengths has been reported. It has been found that both the absorption and scattering efficiencies get blue-shifted with an increase in Au shell layer thickness from 2 to 10 nm and with an increase in surrounding refractive index whereas the corresponding LSPR peaks shift towards red. It has also been found that the spectra are red-shifted with an increase in the core radius from 20 to 40 nm while keeping the shell thickness same. The effect of shell thickness on the absorption peak position and absorption linewidth has also been studied. Hence, the optical response of both CdSe- and CdTe-coated Au nanoshells can be tuned and controlled from the visible to the near-infrared (NIR) region of the electromagnetic (EM) spectrum. Finally, the CdSe-coated Au nanoshell exhibits high scattering and absorption efficiencies in comparison to the CdTe-coated nanoshell.     

压电免疫传感器固定方法

抗原抗体在石英晶体电极表面固定而不丧失其活性是压电免疫传感器成功的关键,直接影响到它的灵敏度和可重复性等性质。介绍了压电免疫传感器传统的表面固定方法:主要有戊二醛交联法、自组装单分子膜法(SAM)、蛋白A固定法;以及国内外最新研究的不需通常固定化步骤的方法,主要是压电凝胶免疫分析法(LEPIA)和PEG压电凝胶分析法,并对这些方法的发展前景做了展望 。     

Immobilization of ovomucoid on chitosan

The inhibitory activity of ovomucoid from duck egg white, immobilized on chitosan with the use of glutaraldehyde or carbodiimide as cross-linking agents, was studied. Glutaraldehyde proved to be a more preferable cross-linking agent than carbodiimide. When chitosan is used as a protein carrier, the possibility of shifting the pH optimum of these compounds should be taken into account.     

A new set up for multi-analyte sensing: at-line bio-process monitoring

A multi-analyte sensing device is described, for simultaneous at-line monitoring of glucose, ethanol, pO?-value and cell density. It consists of a dual biosensor, a modified microscope and a fiber optical pO?-sensor that are integrated into a flow analysis (FA) system. The biosensor is based on a conventional thin layer flow-through cell equipped with a gold (Au) dual electrode (serial configuration). The biosensors with no cross-talking were produced by modifying the electrochemical transducers. Each Au surface was initially modified by self-assembled monolayer (SAM) of cysteamine. Alcohol oxidase (AOx) and pyranose oxidase (PyOx) were immobilized each onto a gold surface by means of PAMAM (polyamidoamine) dendrimer via glutaraldehyde cross-linking. The responses for glucose and ethanol were linear up to 0.5 mM. The operational stability of the biosensors was very promising, after 11 h continuous operation, only 6.0% of the initial activity was lost. The potential of the described biosensor was demonstrated by parallel determination of ethanol and glucose in yeast fermentation process. Simultaneously the cell density of the culture was monitored with an in situ microscope (ISM), which was integrated into the FA system. Both the used in situ microscope and the image processing algorithm used for the analysis of the acquired image data are described. Furthermore the pO?-value was monitored using a fiber optical sensor, which was embedded in a flow cell. The multi-sensor device allows the at-line monitoring of several process values without the need for further sampling or time consuming offline measurements.     

Immobilization of ovomucoid on chitosan

The inhibitory activity of ovomucoid from duck egg white, immobilized on chitosan with the use of glutaraldehyde or carbodiimide as cross-linking agents, was studied. Glutaraldehyde proved to be a more preferable cross-linking agent than carbodiimide. When chitosan is used as a protein carrier, the possibility of shifting the pH optimum of these compounds should be taken into account.     

Mass synthesis of single-crystal gold nanosheets based on chitosan

Single-crystal Au nanosheets with {111} planes as basal surfaces have been synthesized on the basis of the polysaccharide chitosan. The preferential adsorption of polar groups in chitosan molecules on {111} planes of Au nuclei may account for the formation of anisotropic nanosheets. Appropriate precursor (HAuCl(4)) concentrations are vital for the formation of Au nanosheets. The Au nanostructures thus prepared exhibit interesting shape-dependent optical properties. This convenient, environmentally friendly and low-cost route may be amenable to mass production.     

Investigating the Effect of Ag and Au Nanostructures with Spherical and Rod Shapes on the Emission Wavelength of OLED

Noble metals, especially Ag and Au nanostructures, have unique and adjustable optical attributes in terms of surface plasmon resonance. In this research, the effect of Ag and Au nanoparticles with spherical and rod shapes on the light extraction efficiency and the FWHM of OLED structures was investigated using the finite difference time domain (FDTD) method. The simulation results displayed that by changing the shape and size of Ag and Au nanostructures, the emission wavelength can be adjusted, and the FWHM can be reduced. The presence of Ag and Au nanoparticles in the OLEDs showed a blue and red shift of the emission wavelength, respectively. Also, the Ag and Au nanorods caused a significant reduction in the FWHM and a shift to the longer wavelengths in the structures. The structures containing Ag nanorods showed the narrowest FWHM and longer emission wavelength than the other structures.

     

Biocomposite of subtilisin Carlsberg with chitosan as an effective biocatalyst for hydrolysis and synthesis of peptides

New biocatalysts, preparations of subtilisin Carlsberg immobilized on chitosan (a deacetylated derivative of chitin), were obtained. The enzyme content, hydrolytic activity, and ability to catalyze peptide bond formation in organic solvents were characterized for these preparations. The influence of the form and composition of the biocomplosite (content of the enzyme and glutaraldehyde, the cross-linking agent) and buffer pH on the biocatalytic properties of the immobilized enzyme was studied in the reactions of peptide bond hydrolysis. The synthase activity of the preparations was investigated in the reaction of synthesis of Z-Ala-Ala-Leu-Phe-pNA in a 6:4 DMF-acetonitrile mixture in dependence on the reaction time. The yield of this product was 100% after only 40 min.     

Biocomplex of subtilisin Carlsberg with chitosan as an effective biocatalyst for hydrolysis and synthesis of peptides

New biocatalysts, preparations of subtilisin Carlsberg immobilized on chitosan (a deacetylated derivative of chitin), were obtained. The enzyme content, hydrolytic activity, and ability to catalyze peptide bond formation in organic solvents were characterized for these preparations. The influence of the form and composition of the biocomplex (content of the enzyme and glutaraldehyde, the cross-linking agent) and buffer pH on the biocata-lytic properties of the immobilized enzyme was studied in the reactions of peptide bond hydrolysis. The synthase activity of the preparations was investigated in the reaction of synthesis of Z-Ala-Ala-Leu-Phe-pNA in a 6 : 4 DMF-acetonitrile mixture in dependence on the reaction time. The yield of this product was 100% after only 40 min.     

Attachment of ribosome crystals to intracellular membranes.

The attachment to membranes of ribosome crystals formed by cooling lizard oocytes and chick embryos has been investigated by electron microscopy of whole cells and by biochemical and structural experiments, using the cross-linking reagent glutaraldehyde.It was found that the crystalline ribosomes in both animals form only on the rough endoplasmic reticulum and nuclear envelope, that they bind to these membranes through one unique site on the large ribosomal subunit, that the bond between the large subunit and the site on the membrane is sensitive to the concentration of K⁺, but not of Mg²⁺, and that this bond is selectively stabilized by mild treatment with glutaraldehyde. These results closely match those obtained from ribosomes in secretory cells, suggesting that there may be no difference between the two sets of ribosomes in their direct interaction with membranes.The glutaraldehyde reaction was used to obtain crystals and components from which the small subunits had been preferentially released. A comparison between small subunit depleted and normal crystals led to an estimate for the positions of the subunits over the membrane surface. The side-by-side subunit assignments, “S” and “L”, suggested previously (Unwin &; Taddei, 1977; Unwin, 1977), were confirmed. It was deduced further that the crystalline ribosomes have the long axis of their small subunit approximately parallel to the membrane surface, and appear raised up from this surface because of interaction between their large subunits.     

Co-aggregation of enzymes and polyethyleneimine: a simple method to prepare stable and immobilized derivatives of glutaryl acylase

We have developed a novel methodology that allowed the preparation of cross-linked enzyme aggregates (CLEAs) of glutaryl acylase (GAC) by co-aggregation of the enzyme with an aminated polymer: polyethyleneimine (PEI). The preparation of CLEAs of GAC from Pseudomonas sp. is not possible when using poly(ethylene glycol) and glutaraldehyde directly as precipitating and cross-linking agent, respectively. This problem arises probably from the low content of surface Lys groups of GAC which prevents an efficient cross-linking of the enzyme molecules in the aggregate. This fact was proven by the release of enzyme molecules from the aggregate and the solubilization of the enzyme when eliminating the precipitating agent. Our new co-aggregation system favors the cross-linking between the very reactive and abundant primary amino groups of the PEI and the primary amino groups on the enzyme surface. The use of PEI prevents the release of enzyme molecules from the aggregate. By this methodology, we prepared a very stable immobilized derivative of GAC. After optimization of the glutaraldehyde treatment conditions, the stability of the enzyme was significantly improved. It kept more than 60% of its initial activity after 72 h of incubation at 45 degrees C, whereas the soluble enzyme was fully inactivated in 2.5 h of incubation in the same conditions. Therefore, we have a new protocol for carrying out the preparation of cross-linked aggregates of enzymes with a low number of lysines on its surface.     

Cross-linking of dark-adapted frog photoreceptor disk membranes. Evidence for monomeric rhodopsin.

A model for random cross-linking of identical monomers diffusing in a membrane was formulated to test whether rhodopsin's cross-linking behavior was quantitatively consistent with a monomeric structure. Cross-linking was performed on rhodopsin both in intact retinas and in isolated rod outer segment (ROS) membranes using the reagent glutaraldehyde. The distribution of covalent oligomers formed was analyzed by SDS-polyacrylamide gel electrophoresis and compared to predictions for the random model. A similar analysis was made for ROS membranes cross-linked by diisocyanatohexane and retinas cross-linked by cupric ion complexed with o-phenanthroline. Patterns of cross-linking produced by these three reagents are reasonably consistent with the monomer model. Glutaraldehyde was also used to cross-link the tetrameric protein aldolase in order to verify that cross-linking of a stable oligomer, under conditions comparable to those used for ROS, yielded the pattern predicted for a tetrameric protein having D2 symmetry. This pattern is markedly different from the one for a random-collision model. Moreover, a comparison of rates showed that aldolase cross-linking with glutaraldehyde is significantly faster than cross-linking of membrane-bound rhodopsin. It is concluded that rhodopsin is monomeric in dark-adapted photoreceptor membranes and that the observed cross-linking results from collisions between diffusing rhodopsin molecules.