溶胶凝胶技术在生物传感器中的应用

溶胶凝胶过程以基纯度高,均匀性强,处理温度低,反应条件易于控制等优点,已被成功地用以生物组分的固体上。溶胶凝胶技术作为一种颇具前任的固化方法,为生物传感器的制作和发展提供了更多的机会和条件,本文就近几年间溶胶凝胶技术在生物传感器中的开发应用状况进行了综述。     

等离子体增强热丝化学气相沉积法生长取向金刚石薄膜

采用等离子体增强热丝化学气相沉积法在Ｓｉ（１１１）衬底上生长金刚石薄膜,扫描电子显微镜（ＳＥＭ）和Ｘ射线衍射（ＸＲＤ）分析结果表明金刚石晶体颗粒在（１１１）方向实现了取向生长。     

求二维随机变量函数分布的截图降维法

计算二维随机变量函数分布的卷积公式是一个降维公式,通过图形辅助求解,揭示图形中所截的线段实为卷积公式中积分变量的取值范围.以两个实例说明:利用降维公式辅助截图可有效降低计算难度.     

Effects of Alkali Metal Ion Cationization on Fragmentation Pathways of Triazole-Epothilone

The collisionally activated dissociation mass spectra of the protonated and alkali metal cationized ions of a triazole-epothilone analogue were studied in a Fourier transform ion cyclotron resonance mass spectrometer. The fragmentation pathway of the protonated ion was characterized by the loss of the unit of C₃H₄O₃. However, another fragmentation pathway with the loss of C₃H₂O₂ was identified for the complex ions with Na⁺, K⁺, Rb⁺, and Cs⁺. The branching ratio of the second pathway increases with the increment of the size of alkali metal ions. Theoretical calculations based on density functional theory (DFT) method show the difference in the binding position of the proton and the metal ions. With the increase of the radii of the metal ions, progressive changes in the macrocycle of the compound are induced, which cause the corresponding change in their fragmentation pathways. It has also been found that the interaction energy between the compound and the metal ion decreases with increase in the size of the latter. This is consistent with the experimental results, which show that cesiated complexes readily eject Cs⁺ when subject to collisions.     

Global Chemome Study by LC Coupled with DAD and ESI–Q–TOF MS of a Composite Traditional Chinese Medicine Qishenyiqi Dropping Pills

High-performance liquid chromatography coupled with diode-array detection and electrospray ionization quadrupole-time of flight mass spectrometry (ESI–Q–TOF MS) has been used for global chemome analysis of Qishenyiqi dropping pills. Under gradient conditions, two sub-chemomes extracted from Qishenyiqi dropping pills were analyzed on the same C₁₈ reversed-phase column by use of different binary mobile phases. In the chromatogram obtained from sub-chemome I, 19 compounds with strong ultraviolet absorption at 260 nm were detected; 27 compounds with weak ultraviolet absorption at 203 nm were detected in the chromatogram obtained from sub-chemome II. All the compounds were identified by ESI–Q–TOF MS. This study furnished much more important information about the pharmacological activity of Qishenyiqi dropping pills than previous studies and laid a good foundation for further exploration of chemomics.     

Bioactivity‐integrated ultra‐performance liquid chromatography/quadrupole time‐of‐flight mass spectrometry for the identification of nuclear factor‐κB inhibitors and β2 adrenergic receptor agonists in Chinese medicinal preparation Chuanbeipipa dropping pills

A simple and dual‐target method based on ultra‐performance liquid chromatography/quadrupole time‐of‐flight mass spectrometry combined with dual‐bioactive [nuclear factor‐κB (NF‐κB) and β₂‐adrenergic receptor] luciferase reporter assay systems was developed to rapidly characterize the chemical structure of various bioactive compounds of TCM preparations. Chuanbeipipa dropping pills, a traditional Chinese medicine preparation used for the clinical therapy of chronic obstructive lung disease and cough caused by bronchial catarrh, was analyzed with this method. Potential anti‐inflammatory and spasmolytic constituents were screened using NF‐κB and β₂‐adrenergic receptor activity luciferase reporter assay systems and simultaneously identified according to the time‐of‐flight mass spectrometry data. One β₂‐adrenergic receptor agonist (ephedrine) and two structural types of NF‐κB inhibitors (platycosides derivatives and ursolic acid derivatives) were characterized. Platycodin D3 and E were considered new NF‐κB inhibitors. Further cytokine and chemokine detection confirmed the anti‐inflammatory effects of the potential NF‐κB inhibitors. Compared with conventional fingerprints, activity‐integrated fingerprints that contain both chemical and bioactive details offer a more comprehensive understanding of the chemical makeup of plant materials. This strategy clearly demonstrated that multiple bioactivity‐integrated fingerprinting is a powerful tool for the improved screening and identification of potential multi‐target lead compounds in complex herbal medicines. Copyright © 2013 John Wiley & Sons, Ltd.     

Improving the Thermostability of a Methyl Parathion Hydrolase by Adding the Ionic Bond on Protein Surface

The thermostability of the methyl parathion hydrolase (MPH_OCH) from Ochrobactrum sp. M231 was improved using site-directed mutagenesis. Two prolines (Pro76 and Pro78) located on the protein surface were selected for mutations after inspection of the sequence alignment of MPH_OCH and OPHC2, a thermostable organophosphorus hydrolase from Pseudomonas pseudoalcaligenes C2-1. The temperature of the double-point mutant (P76D/P78K) at which the mutant lost 50% of its activity (T50) was approximately 68 °C, which is higher than that of WT enzyme (64 °C), P76D (67 °C), and P78K (59 °C). Structural analysis of P76D/P78K indicated that the substituted residues (Asp76 and Lys78) could generate an ionic bond and increase the structural electrostatic energy, which could then increase the stability of the protein. These results also suggest that the thermal stability of proteins could be improved by adding the ionic bond on protein surface.     

A metabonomic approach applied to predict patients with cerebral infarction

Cerebral infarction is always of sudden onset, and usually leading to serious consequence. It is of therapeutic significance to develop fast and accurate diagnosis methods for cerebral infarction so that patients can be treated timely and properly. A metabonomic approach was then proposed to investigate the potential biomarkers and metabolic pathways associated with cerebral infarction and also establish a prediction model of cerebral infarction for the fast diagnosis. Serum metabolic profiling of sixty-seven cerebral infarction patients and sixty-two controls was obtained using UPLC-TOF MS. The resulting data were then processed by multivariate statistical analysis to graphically demonstrate metabolic variations. The PLS-DA model was validated with cross validation and permutation tests to assure the model's reliability, and significant difference was obtained between the original and hypothetical models (p < 0.0001). A series of endogenous metabolites in the one-carbon cycle, such as folic acid, cysteine, S-adenosyl homocysteine and oxidized glutathione, were determined as potential biomarkers of cerebral infarction. A prediction model developed using PLS-KNN algorithm was established to differentiate cerebral infarction patients from controls, and an average accuracy of 100% was obtained. In conclusion, metabonomic approach is a powerful tool to investigate the pathogenesis of stroke and is expected to be developed as a useful method for the fast diagnosis.