失效原子力显微镜硅针尖再生

原子力显微镜的传统商品硅针尖在使用过程中极易因磨损而失效，本文研究了一种在实验室条件下简易可行的回收利用失效硅针尖的方法。在原子力显微镜的敲击模式下使用曲率半径大于100 nm的失效硅针尖对生长单壁碳纳米管的样品表面进行扫描，把样品表面的单壁碳纳米管管束粘接到硅针尖上，可制得直径在5～20 nm的碳纳米管针尖。实验对碳纳米管针尖和新的商品硅针尖进行了成像对比，所制备的碳纳米管针尖不仅在成像分辨率而且在成像稳定性上都优于新的商品硅针尖。     

“学习提示”模块在新版有机化学教材中的应用

一本好的有机化学教材应该对教师教学和学生自学有很好的辅助和引导作用。如何使学生从浩如烟海的内容中发现重点、难点,并做到有效掌握和融会贯通,是教材编写者需要重点考虑的问题。笔者将教学经验融入到有机化学教材改革实践,在新版教材中加入了原创的“学习提示”板块,用简练、更通俗易懂的语言对一些重要、较难理解的或是容易混淆的知识点进行辅助讲解、串联总结。笔者认为该板块可以起到协助学生理解知识难点、厘清易混淆概念、将关联知识融会贯通以及激发学生科研兴趣等作用。本文用一些实例对这一模块的编写构思、内容和预期成效进行了介绍。     

In-tip nanoreactors for cancer cells proteome profiling

Mass spectrometry (MS)-based proteome profiling is essential for molecular diagnostics in modern biomedical study. To date, sample preparation including protein extraction and proteolysis is still very challenging and lack of efficiency. Recently tips-based sample preparation protocols exhibit strong potentials to achieve the goal of “a proteome in an hour”. However, in-tip proteolysis is still rarely reported and far from ideal for dealing with complex bio-samples. In this work, nanoreactors encapsulated micropipette tips were demonstrated as high performance devices for fast (∼minutes) and multiplexing proteolysis to assist the profiling of cancer cells proteome. Nanoporous silica materials with controlled pore size and surface chemistry were prepared as nanoreactors and encapsulated in micropipette tips for efficient in situ proteolysis. The as-constructed device showed desirable sensitivity (LOD of 0.204 ± 0.008 ng/μL and LOQ of 0.937 ± 0.055 ng/μL), selectivity, stability (two months under −20 °C), reusability (at least 10 times), and little memory effect in MS based bottom-up proteomic analysis. It was used for comprehensive protein mapping from cancer cell lines. The number of identified proteins was increased by 18%, 22%, 52%, and 52% dealing with HepG2, F56, MCF7, and HCCLM3 cancer cells, compared to traditional in-solution proteolysis based bottom-up proteomic strategy. With the enhanced performance, our work built a novel, efficient and miniaturized platform for facile proteomic sample preparation, which is promising for advanced biomarkers discovery in biomedical study.     

Direct analysis of herbal powders by pipette-tip electrospray ionization mass spectrometry

Conventional electrospray ionization mass spectrometry (ESI-MS) is widely used for analysis of solution samples. The development of solid-substrate ESI-MS allows direct ionization analysis of bulky solid samples. In this study, we developed pipette-tip ESI-MS, a technique that combines pipette tips with syringe and syringe pump, for direct analysis of herbal powders, another common form of samples. We demonstrated that various herbal powder samples, including herbal medicines and food samples, could be readily online extracted and analyzed using this technique. Various powder samples, such as Rhizoma coptidis, lotus plumule, great burdock achene, black pepper, Panax ginseng, roasted coffee beans, Fructus Schisandrae Chinensis and Fructus Schisandrae Sphenantherae, were analyzed using pipette-tip ESI-MS and quality mass spectra with stable and durable signals could be obtained. Both positive and negative ion modes were attempted and various compounds including amino acids, oligosaccharides, glycosides, alkaloids, organic acids, ginosensides, flavonoids and lignans could be detected. Principal component analysis (PCA) based on the acquired mass spectra allowed rapid differentiation of closely related herbal species.     

Quantitative determination of phenothiazine derivatives in human plasma using monolithic silica solid-phase extraction tips and gas chromatography-mass spectrometry

Solid-phase extraction (SPE) using micropipette tips is a useful technique to prepare samples prior to mass spectrometry. However, most commercial SPE tips have loading capacities that are insufficient for quantitative determination. In this paper, we describe a rapid method for quantitative microanalysis of five phenothiazine derivatives, chlorpromazine, levomepromazine, promazine, promethazine and trimeprazine, using a recently introduced C(18) monolithic silica SPE tip, the MonoTip C(18), for extraction from human plasma. The drugs could be extracted within 5 min from 0.1-mL plasma samples, eluted with methanol, and the eluate injected directly into a gas chromatograph prior to mass spectrometry analysis. Only 0.7 mL of solvent was required for each step of the extraction process. The recoveries of the five phenothiazines spiked into plasma were 91-95% and the limits of quantification for each drug were between 0.25 and 2.0 ng/0.1 mL. The maximum intra- and inter-day coefficient of variation was 11%. The validated method was successfully used to quantify the plasma concentration of levemepromazine in a human subject after oral administration of the drug. This new method is expected to have wide applications as a pretreatment for the rapid, quantitative determination of drug concentrations in plasma samples.     

果胶甲基酯酶PME参与调控大麦根尖铝毒敏感性

利用大麦耐铝品种和敏感品种来分析根尖果胶甲基酯酶（PME）在大麦铝毒敏感性中的调控作用.通过对耐铝性差异极其显著的2个大麦明品种2000-2与沪麦16经铝处理后的Morin荧光检测,观察到2000-2根尖荧光明显比沪麦16强,铝试剂比色法测定结果进一步表明,敏感品种根尖细胞壁上铝积累量比耐铝品种高,达到极显著差异（p〈0.05或0.01）.对根尖PME活性分析结果表明,与对照相比,铝处理4 h的2000-2根尖PME活性显著上升,但处理24 h后PME活性又显著下降;而沪麦16的PME活性与对照相比无显著性差异.这些结果表明细胞壁上铝积累与PME活性变化有密切的相关性,证实PME活性参与调控大麦铝毒敏感性.     

血细胞压积与平均血红蛋白浓度的可见-近红外光谱无创检测

血细胞压积和平均血红蛋白浓度的检测对于心脑血管疾病以及贫血等疾病的预防和治疗起着非常重要的作用,为了能够无创、准确的对血细胞压积和平均血红蛋白浓度进行检测,本研究提出了一种基于光谱技术的检测方法。经解剖学研究表明,在微循环中,血流变的异常以及血液粘稠度的改变均能引起舌象的变化,即舌象和血液成分之间存在一定的相关性,所以本研究通过采集240例志愿者舌尖反射光谱,同时拍摄志愿者舌体图片和记录生化分析结果。在进行数学建模前,首先将240例志愿者随机分为校正集和验证集,运用偏最小二乘分别建立数学模型,校正集预测值和生化分析真实值之间相关系数分别为0.998和0.938,运用所建数学模型对验证集未知样本进行预测,验证集未知样本的预测结果与生化分析真实值之间相关系数分别为0.979和0.883,平均相对误差分别为1.65%和1.88%,均方根误差分别为4.066和4.139,由实验结果可见,所建立的模型可以较好的对HCT和MCHC进行预测,同时实验结果也表明近红外光谱分析结合偏最小二乘方法有可能能够为血细胞压积和平均血红蛋白浓度的检测提供一种无创的检测方法。     

An effective and in-situ method based tresyl-functionalized porous polymer material for enrichment and digestion of membrane proteins and its application in extraction tips

Membrane proteins are one of promising targets for drug discovery because of the unique properties in physiological processes. Due to their low abundance and extremely hydrophobic nature, the analysis of membrane proteins is still a great challenge. In this work, an effective and in-situ method were developed to enrich and digest membrane proteins by adopting tresyl-functionalized porous polymer material. With tresyl groups, the material can effectively immobilize membrane proteins via covalent bonding on the surface. The material became a facile carrier to enrich membrane proteins from the rat liver in detergents and organic solvents owing to its outstanding binding capacity and excellent biocompatibility. Moreover, it was further applied in extraction tips to capture and in-situ digest the pretreatment membrane proteins in two different solutions. A total of 600 membrane proteins (51% of total protein groups) and 359 transmembrane proteins were identified by nano-LC-ESI-MS/MS in 4% sodium dodecyl sulfate (SDS), and similar results were achieved in the 60% methanol solution. All these results demonstrated that the new approach is of great promise for large-scale characterization of membrane proteins.     

STM和ECSTM探针的制备及探针对图象的影响

本文综述了扫描隧道显微镜(STM)以及现场电化学扫描隧道显微镜(ECSTM)探针的制备方法,并对相应探针制备方法的优缺点以及探针性能对图象影响方面的进展进行了评述。     

Single wall carbon nanotubes and their electrical properties

Single-wall carbon nanotubes (SWCNTs) were synthesized and purified. A water colloid of SWCNTs was prepared and used to assemble SWCNTs onto a gold film surface. Scanning tunneling microscopy (STM) images showed that short SWCNTs stood on gold film surfaces. Using STM tips made of SWCNTs, a crystal grain image of a gold thin film and an atomic resolution image of highly oriented pyrolytic graphite were successfully obtained. The electrical properties of short SWCNTs, which stood on the surface of gold film, were measured using STM. That SWCNTs stand on gold thin films is a promising technique for studying structures and properties of carbon nanotubes, as well as assembling and fabricating high-intensity coherent electron sources, field emission flat panel display, tips for scanning probe microscopes, new nanoelectronic devices, etc.