基因治疗用质粒DNA的制备工艺

目的探索基因治疗用质粒DNA的制备工艺。方法以CaCl2沉淀法去除大分子RNA,Q-Sepharose和SOURCE两步离子交换层析法去除染色体DNA、小分子RNA、蛋白质等杂质,并对各步骤样品进行检测。结果质粒pIRVP3HNIL18终产品的产量为1.638mg质粒DNA/g细菌,纯度为1.839,相对回收率为12.55%,蛋白质含量为0.0028mg/ml,未检测到RNA和染色体DNA。结论此工艺可有效去除质粒DNA制备过程中产生的杂质,可应用于药剂水平质粒DNA的制备。     

Solid State Viscoelastic Properties,Morphological and Melt Rheological Studies on PLA/TPU/POSS Nanocomposites

Effects of trifluoropropyl-substituted polyhedral oligomeric Silsesquioxane (POSS), on the morphological, rheological and dynamic mechanical properties of a series of poly(lactic acid)

(PLA)/thermoplastic polyurethane (TPU) blends were explored, thoroughly. Microscopic techniques including scanning electron microscope (SEM) and transmission electron microscope (TEM) utilized in conjunction with viscosity measurements to explain morphological dissimilarities between different samples. A remarkable morphological refinement was observed upon addition of POSS into the 50/50 PLA/TPU blend. This morphological change was detected by means of frequency sweep rheological experiments, where the shoulder-like plateau appears as a result of increase in the contribution from the interfacial component of elasticity in the enhancement of storage modulus. Based on the results of the dynamic time sweep small amplitude oscillatory shear (SAOS) experiments, a scaling relation was proposed to quantify the kinetics of phase separation and the associate time evolution of elasticity. It turned out that POSS particles can slow down the rate of morphological coarsening up to the 50%. Using dynamic mechanical thermal analysis (DMTA) and rheological experiments, the differences in the rate of undergoing dynamics in all the length scales, starting from segmental up to the chain level, were compared for various compositions and the discrepancies were fully described based on the dissimilarities in the entanglement densities, combinatorial effects of soft and hard segments of TPU, lubricating role of POSS particles, crystallinity, and morphological characteristics of samples.     

小鼠神经生长因子基因治疗型DNA质粒的质量控制

目的对小鼠神经生长因子(NGF)基因治疗型DNA质粒进行质量控制。方法用酶切鉴定法和PCR法进行DNA质粒的结构确认,鸡胚背根神经节法和免疫印迹测定DNA质粒表达产物的生物学活性,分光光度法测定浓度,琼脂糖凝胶电泳法和DNA-NPR-HPLC法测定纯度,气相色谱法测定乙醇和异丙醇残留量,琼脂糖凝胶电泳法测定RNA残留量,其余检测项目按《中国药典》三部(2005版)规定进行。结果用上述方法对原液和成品进行了检定,各项指标均符合《预防用DNA疫苗临床前研究技术指导原则》和《中国药典》三部(2005版)的要求。结论所采用的质控方法和质量标准能够保证该DNA质粒的安全、有效,可用于治疗型DNA质粒的质控。     

Microstructural and Optoelectronic Studies on Polyaniline: TiO2 Nanocomposites

Thin films of polyaniline (PANi) and PANi: titanium oxide (TiO₂) composites have been synthesized by sol–gel spin coating technique. The TiO₂ powder of particle size 50–60 nm was synthesized by the sol–gel technique and the polyaniline was synthesized by the chemical oxidative polymerization of aniline. The composite films were characterized by X-ray diffraction (XRD), atomic force microscopy (AFM), scanning electron microscopy (SEM), Fourier transform infrared (FTIR) spectroscopy, UV-vis spectroscopy and the four-probe method. The results were compared with corresponding data on pure polyaniline films. The intensity of diffraction peaks for PANi:TiO₂ composites is lower than that for TiO₂. The characteristic FTIR peaks of pure PANi are observed to shift to a higher wavenumber in PANi:TiO₂ composite, which is attributed to the interaction of TiO₂ particles with PANi molecular chains. The resistivity measurement shows that the molecular chain constitution of polyaniline is the most important carrier in the polyaniline: nano-TiO₂ composite.     

PVA,PVA Blends,and Their Nanocomposites for Biodegradable Packaging Application

This review exclusively addresses material systems primarily based on poly(vinyl alcohol) (PVA), one of the most popular water-soluble biopolymers, for their use in packaging applications with the primary objective of reducing petrobased plastic waste. In addition, some typical PVA blends and nanocomposites are discussed as comparative studies for material packaging. Structural characteristics, mechanical, thermal, and barrier properties, in addition to biodegradation of these multiple material systems are summarized in a systematic manner. Finally, associated fabrication processing methods together with the most popular theoretical models used for the permeability of PVA nanocomposites are also reviewed in detail.     

Increased Circulatory Asymmetric Dimethylarginine and Multiple Organ Failure: Bile Duct Ligation in Rat as a Model

Bile duct ligation (BDL)-treated rats exhibit cholestasis, increased systemic oxidative stress, and liver fibrosis, which ultimately lead to liver cirrhosis. Asymmetric dimethylarginine (ADMA) is a competitive inhibitor of nitric oxide synthase that can decrease the synthesis of nitric oxide. BDL rats have higher plasma and hepatic ADMA levels, which may be due to increased hepatic protein arginine methyltransferase-1 and decreased dimethylarginine dimethylaminohydrolase expression. BDL rats also exhibit renal and brain damage characterized by increased tissue ADMA concentrations. The increased plasma ADMA levels and multiple organ damages seen here are also observed following multiple organ failures associated with critical illness. This review discusses the dysregulation of ADMA in major organs in BDL rats and the role of increased ADMA in multiple organ damages.     

Studies on Preparation of Photosensitizer Loaded Magnetic Silica Nanoparticles and Their Anti-Tumor Effects for Targeting Photodynamic Therapy

Abstract  As a fast developing alternative of traditional therapeutics, photodynamic therapy (PDT) is an effective, noninvasive, nontoxic therapeutics for cancer, senile macular degeneration, and so on. But the efficacy of PDT was compromised by insufficient selectivity and low solubility. In this study, novel multifunctional silica-based magnetic nanoparticles (SMNPs) were strategically designed and prepared as targeting drug delivery system to achieve higher specificity and better solubility. 2,7,12,18-Tetramethyl-3,8-di-(1-propoxyethyl)-13,17-bis-(3-hydroxypropyl) porphyrin, shorted as PHPP, was used as photosensitizer, which was first synthesized by our lab with good PDT effects. Magnetite nanoparticles (Fe₃O₄) and PHPP were incorporated into silica nanoparticles by microemulsion and sol–gel methods. The prepared nanoparticles were characterized by transmission electron microscopy, X-ray diffraction, Fourier transform infrared spectroscopy and fluorescence spectroscopy. The nanoparticles were approximately spherical with 20–30 nm diameter. Intense fluorescence of PHPP was monitored in the cytoplasm of SW480 cells. The nanoparticles possessed good biocompatibility and could generate singlet oxygen to cause remarkable photodynamic anti-tumor effects. These suggested that PHPP-SMNPs had great potential as effective drug delivery system in targeting photodynamic therapy, diagnostic magnetic resonance imaging and magnetic hyperthermia therapy. Graphical Abstract   Novel multifunctional photosensitizer loaded magnetic silica nanoparticles were strategically prepared with low toxicity, good biocompatibility and remarkable photodynamic anti-tumor efficacy. The nanoparticles were believed to be of great value as drug delivery system in targeting photodynamic therapy, diagnostic magnetic resonance imaging and magnetic hyperthermia therapy.      

Synthetic Nucleic Acid Analogues in Gene Therapy: An Update for Peptide–Oligonucleotide Conjugates

The main objective of this work is to provide an update on synthetic nucleic acid analogues and nanoassemblies as tools in gene therapy. In particular, the synthesis and properties of peptide–oligonucleotide conjugates (POCs), which have high potential in research and as therapeutics, are described in detail. The exploration of POCs has already led to fruitful results in the treatment of neurological diseases, lung disorders, cancer, leukemia, viral, and bacterial infections. However, delivery and in vivo stability are the major barriers to the clinical application of POCs and other analogues that still have to be overcome. This review summarizes recent achievements in the delivery and in vivo administration of synthetic nucleic acid analogues, focusing on POCs, and compares their efficiency.     

用于基因转运和治疗的多功能纳米复合物的制备及应用

基因治疗是将DNA转染进入目的细胞,修复遗传错误或者产生治疗因子。目前基因治疗研究的热点是利用纳米颗粒作为载体,将特异性基因转染到细胞内,从而达到基因治疗的目的。纳米颗粒作为载体可使基因的转染效率提高、增强其稳定性、靶向型。就纳米复合物的组成及在基因治疗方面的应用、面临的问题与挑战进行了综述。     

A Single Gene Cluster for Chalcomycins and Aldgamycins: Genetic Basis for Bifurcation of Their Biosynthesis

Aldgamycins are 16‐membered macrolide antibiotics with a rare branched‐chain sugar d ‐aldgarose or decarboxylated d ‐aldgarose at C‐5. In our efforts to clone the gene cluster for aldgamycins from a marine‐derived Streptomyces sp. HK‐2006‐1 capable of producing both aldgamycins and chalcomycins, we found that both are biosynthesized from a single gene cluster. Whole‐genome sequencing combined with gene disruption established the entire gene cluster of aldgamycins: nine new genes are incorporated with the previously identified chalcomycin gene cluster. Functional analysis of these genes revealed that almDI/almDII, (encoding α/β subunits of pyruvate dehydrogenase) triggers the biosynthesis of aldgamycins, whereas almCI (encoding an oxidoreductase) initiates chalcomycins biosynthesis. This is the first report that aldgamycins and chalcomycins are derived from a single gene cluster and of the genetic basis for bifurcation in their biosynthesis.     

Soft X‐ray Ptychographic Imaging and Morphological Quantification of Calcium Silicate Hydrates (C–S–H)

Morphological details of calcium silicate hydrate (C–S–H) stemming from the hydration process of Portland cement (PC) phases are crucial for understanding the PC‐based systems but are still only partially known. Here we introduce the first soft X‐ray ptychographic imaging of tricalcium silicate (C₃S) hydration products. The results are compared using both scanning transmission X‐ray and electron transmission microscopy data. The evidence shows that ptychography is a powerful method to visualize the details of outer and inner product C–S–H of fully hydrated C₃S, which have fibrillar and an interglobular structure with average void sizes of 20 nm, respectively. The high‐resolution ptychrography image enables us to perform morphological quantification of C–S–H, and, for the first time, to possibly distinguish the contributions of inner and outer product C–S–H to the small angle scattering of cement paste. The results indicate that the outer product C–S–H is mainly responsible for the q^?3 regime, whereas the inner product C–S–H transitions to a q^?2 regime. Various hypotheses are discussed to explain these regimes.     

Metallo-supramolecular Polymers: Versatile DNA Binding and Their Cytotoxicity

Metallo-supramolecular polymers (RuL1, RuL2, RuL3, FeL3, and FeL4) prepared by complexation of bis(terpyridine) derivatives with Ru²⁺ or Fe²⁺ ions with octahedral coordination structures showed high binding ability to several DNAs (calf thymus DNA, herring sperm DNA, [poly(dA-dT)]₂, and [poly(dG-dC)]₂), which were revealed by UV–Vis absorption titration experiments. The electrostatic interactions between the metal cations of the polymers and phosphate anions of DNA led to formation of conjugate structure. The binding constant observed reached 3.7 × 10⁷ M^?1, which is the highest among values reported for metal complexes to date. Based on a long strand structure of the polymer, groove binding is most possible binding mode. Cell viability experiments showed that RuL3 and FeL3 displayed highly statistical significance (**p<0.01) to human non small cell lung cancer cell lines (NCI-H460).     

The Impact of Zeta Potential and Physicochemical Properties of TiO2‐Based Nanocomposites on Their Biological Activity

The aim of this research was to observe the relationship between zeta potential, morphology, surface area, porosity, chemical composition, and ecotoxicity of nanocomposite powders such as Au/TiO₂, Ag₂O/TiO₂, PdO/TiO₂, Ag/TiO₂/SiO₂, Ag/N(C)TiO₂, and SiO₂/TiO₂ from which Ag₂O/TiO₂, Ag/N(C)/TiO_2, and Ag/TiO₂/SiO₂ were exhibiting good antimicrobial properties. It was observed, that nanomaterials characterized by similar morphology and zeta potential revealed the similar toxic behavior. The samples of higher agglomeration and higher zeta potential, especially Ag/TiO₂/SiO₂ xerogel and TiO₂/SiO₂ aerogel were generally less ecotoxic to water organisms and plants. They were also not genotoxic in concentrations up to 500 and 250 mg/L, respectively.     

色谱分离动力学的理论模型及其应用

色谱模型是研究色谱分离过程的基础,对于描述该过程中质量传递动力学和吸附过程具有非常重要的帮助。本文对近年来色谱分离动力学研究常用理论模型的发展进行了综述,重点介绍了普遍化速率模型、集总孔扩散模型、平衡扩散模型和传递扩散模型4种常用的色谱模型,并讨论了各模型的优缺点及应用范围。     

Modern Template Design and Biological Evaluation of Cephradine-loaded Magnesium Calcium Silicate Nanocomposites as an Inhibitor for Nosocomial Bacteria in Biomedical Applications

Silicon - Magnesium calcium silicate nanostructures (MCSNS) loaded with (0.0, 0.6, 0.9, and 1.2 wt%) of Cephradine-drug consisting of mesoporous particles were functionally prepared by sol-gel...