Solvent-dependent intramolecular charge transfer dual fluorescence of p-dimethylaminobenzanilide bearing steric ortho,ortho-dimethyl substituents at amido aniline

Intramolecular charge transfer (ICT) dual fluorescence was observed in various organic solvents with p-dimethylaminobenzanilide (DMBA) derivatives bearing ortho-methyl (DMOMBA) and ortho,ortho-dimethyl (DMDMBA) substituents at amido aniline moiety. Ab initio calculation and absorption spectral data indicated that high steric hindrance was introduced by the ortho,ortho-dimethyl substitutions. It was found that, with DMDMBA, the CT emission initially shifted to the red with increasing solvent polarity from cyclohexane (CHX, 480 nm) to diethyl ether (DEE, 520 nm), similar to those of DMBA derivatives with the ortho-, meta- or para-methyl substitutions at amido aniline moiety. However, there is a characteristic blue-shift of the long wavelength emission between DEE and tetrahydrofuran (THF, 424 nm) then a bathochromic shift again in highly polar solvent acetonitrile (ACN, 484 nm). The unusual solvent-dependent CT emission was ascribed to two competitive CT channels. One is benzanilide (BA)-like CT, whose CT reaction occurs from amido aniline to benzoyl moiety in nonpolar solvent CHX and DEE; the other one is p-dimethylaminobenzamide (DMABA)-like, whose CT reaction occurs from dimethylamino to benzanilide moiety in highly polar solvent THF and ACN. These findings revealed the steric effect plays an important role in the ICT process, which may alter the properties of the electron donor and/or acceptor, but also change the reaction potential.     

Determination of trace carvedilol by solid substrate–room temperature phosphorimetry,based on its activating effect on hypochlorite‐oxidizing amaranth using sodium dodecyl benzene sulphonate as sensitizer

This work proposes a simple and sensitive solid substrate–room temperature phosphorimetry (SS–RTP) for the selective determination of carvedilol (CV). The method is based on the sensitizing effect of sodium dodecyl benzene sulphonate (SDBS) on CV to activate the oxidation between NaClO and amaranth, resulting in the intense quenching of room temperature phosphorescence (RTP) of the system. Compared with non‐SDBS system, the reduction of phosphorescence intensity (ΔI_p) with SDBS is 16.5 times higher and is directly proportional to the content of CV, covering a wide range 0.080–16.00 fg/spot. The regression equation of the working curve can be expressed as ΔI_p = 0.7780 + 7.057 m_CV (fg/spot) (correlation coefficient (r) = 0.9976, n = 8), with a detection limit (LD) of 0.020 fg/spot (corresponding concentration is 5.1 × 10⁻¹⁴ g/mL, sample volume is 0.40 μL/spot). This sensitive method has also been applied to determine trace CV in human plasma and the results agreed with synchronous fluorimetry (SF). The activation energy (E) and rate constant (k) of this activating reaction were 69.04 kJ/mol and 3.580 × 10⁻⁴ s⁻¹, respectively. The reaction mechanism is also discussed. Copyright © 2011 John Wiley & Sons, Ltd.     

葡聚糖磁性纳米颗粒外加磁场对人树突状细胞基因转染效率的研究

目的研究葡聚糖磁性纳米颗粒（the dextran coated magnetic iron oxide nanoparticles,DMN）在外加钕一铁一硼稀土固定磁场的作用下对人树突状细胞转染效率以及安全性的影响。方法先通过磁力计对DMN进行分析;再将修饰有多聚赖氨酸（Poly-L—Lysine,PLL）的DMN携带绿色荧光蛋白pEGFP—Cl质粒报告基因,在钕-铁-硼稀土周定强磁场的作用下,体外转染人树突状细胞,用荧光显微镜直接观察和流式细胞仪检测来评价外加磁场对DMN作为人树突状细胞转染载体效率的影响;在转染后采用MTT比色法测定在磁场干预下的DMN对人树突状细胞增殖和功能的影响以了解其细胞毒性。结果DMN的核心直径〈30nm,具有明硅的超顺磁性,比饱和磁化强度也明显高于相同Fe3O4含量的普通磁块;DMN作为基因载体在外加磁场作用下,转染12h即可将报告基因转染至人树突状细胞内并成功表达,在荧光显微镜下可观察到绿色荧光细胞,24h转染率可达到最高（约为27％）,转染效率较未加磁场组提高了2～4倍。而且转染后的人树突状细胞增殖活性及功能未因DMN外加磁场及其作用时间的长短而受到影响。结论超顺磁性的DMN在外加磁场作用下可以明显、安全、有效地提高对人树突状细胞的转染效率。     

壳聚糖微球分散体系的抑菌性研究

以乳化交联法制备的壳聚糖微米微球(CMs)为试验材料,研究其对金黄色葡萄球菌和大肠杆菌的抑菌行为,并对其浓度因素、粒径因素和抑菌机理做了探讨。结果表明CMs的抑菌作用随其浓度的增加和粒径的减小而增强。在浓度<0.7 mg/mL时,CMs对细菌的生长具有先抑制后促进的作用;当浓度≥0.7 mg/mL时对细菌有杀灭作用。CMs对两种细菌的最小抑菌浓度和最小杀菌浓度分别为0.8、1.4 mg/mL和1.0、1.8 mg/mL,表明壳聚糖微球对金黄色葡萄球菌和大肠杆菌均具有明显的抑制作用,且金黄色葡萄球菌对微球抑制作用的反应比大肠杆菌更敏感。扫描电子显微镜观察发现,细菌能迅速贴附在CMs表面,并且贴壁后的细菌发生了明显的形态变化,有的甚至完全被破坏。     

Comparative proteogenomic analysis of the Leptospira interrogans virulence-attenuated strain IPAV against the pathogenic strain 56601

The virulence-attenuated Leptospira interrogans serovar Lai strain IPAV was derived by prolonged laboratory passage from a highly virulent ancestral strain isolated in China. We studied the genetic variations of IPAV that render it avirulent via comparative analysis against the pathogenic L. interrogans serovar Lai strain 56601. The complete genome sequence of the IPAV strain was determined and used to compare with, and then rectify and reannotate the genome sequence of strain 56601. Aside from their highly similar genomic structure and gene order, a total of 33 insertions, 53 deletions and 301 single-nucleotide variations (SNVs) were detected throughout the genome of IPAV directly affecting 101 genes, either in their 5' upstream region or within their coding region. Among them, the majority of the 44 functional genes are involved in signal transduction, stress response, transmembrane transport and nitrogen metabolism. Comparative proteomic analysis based on quantitative liquid chromatography (LC)-MS/MS data revealed that among 1 627 selected pairs of orthologs, 174 genes in the IPAV strain were upregulated, with enrichment mainly in classes of energy production and lipid metabolism. In contrast, 228 genes in strain 56601 were upregulated, with the majority enriched in the categories of protein translation and DNA replication/repair. The combination of genomic and proteomic approaches illustrated that altered expression or mutations in critical genes, such as those encoding a Ser/Thr kinase, carbon-starvation protein CstA, glutamine synthetase, GTP-binding protein BipA, ribonucleotide-diphosphate reductase and phosphate transporter, and alterations in the translational profile of lipoproteins or outer membrane proteins are likely to account for the virulence attenuation in strain IPAV.     

Effects of cadmium on carbohydrate and protein metabolisms in the freshwater crab Sinopotamon yangtsekiense

The physiological impact of Cd(2+) on Sinopotamon yangtsekiense was evaluated through changes of selected parameters considered as key elements of carbohydrate and protein metabolisms. Crab were exposed to 0.725, 1.45, 2.9mg·L(-1) Cd(2+) for 7, 14 and 21 days. A time- and/or concentration- dependent decrease in muscle glycogen and increase in LDH activity suggested that glycolysis was accelerated during the treatments. Increased protease activity, lowering of FAA and the initially increased and subsequently decreased aminotransferase activities suggest an enhanced protein mobilization during early Cd(2+) exposure followed by a metabolic impairment during late exposure. Decreased hemolymph glucose level was observed in the crabs treated with 2.9mg·L(-1) Cd(2+) for 21d, suggesting an impaired gluconeogenesis. Ammonia level barely changed during the 14d Cd(2+) exposure most likely due to the increased urea and glutamine production; After 1.45 and 2.9mg·L(-1) Cd(2+) treatment for 21d, ammonia was observed increased followed by an exclusive increase in glutamine. Taken together, our results indicate that carbohydrate and protein are mobilized to a varying degree as a compensatory metabolism to response to the energy stress during acute Cd(2+) exposure. As the time lapsed, some symptoms on metabolism obstacle reflect the toxic effect of sublethal Cd(2+).     

人CD24分子成熟多肽的原核表达及抗体制备

目的: 为获得抗人CD24分子成熟多肽核心蛋白(hCD24N)的多克隆抗体。方法: 制备CD24表达阳性的人肿瘤细胞cDNA,采用PCR法扩增hCD24N的编码基因,构建pGEX-KGV-hCD24N原核表达质粒;转化大肠杆菌BL21(DE3),乳糖诱导表达;经GST亲和柱层析、SDS-PAGE和Western blotting制备并鉴定纯化的GST-StraptagII-hCD24N融合蛋白;免疫新西兰大白兔制备抗血清并用rProtein A亲和柱层析纯化多克隆IgG抗体;用间接ELISA法测定抗体效价,Western blotting鉴定抗体特异性,同时采用细胞免疫荧光检测技术对抗体的特异性和应用可行性作进一步评价。结果: 实现了hCD24N基因的克隆以及在原核细胞中的可溶性重组融合表达,得到了纯化后的目的融合蛋白,并以其为免疫原获得了效价高于1:100 000的抗hCD24N多克隆抗体,Western blotting及细胞免疫荧光检测证明该抗体与当前市售的抗人CD24抗体具有相似的免疫反应特异性,并且能够与CD24阳性人肿瘤细胞表达并加工的高度糖基化CD24天然分子发生特异性抗原-抗体反应。结论: 抗hCD24N多克隆抗体的成功制备为进一步以CD24分子为靶点的肿瘤生物学基础研究以及相关癌症的诊断试剂开发奠定基础。     

双核素心肌显像和门控心肌灌注显像检测糖尿病心肌病的临床应用

目的:采用一日法静息99mTc-甲氧基异丁基异腈(MIBI)及18 F-脱氧葡萄糖(FDG)双核素同时采集(DISA)心肌灌注代谢显像与门控心肌灌注显像,评价糖尿病心肌病(DCM)患者心肌灌注、存活和心功能状况。方法:36例临床诊断为DCM患者,进行99mTc-MIBI和18F-FDG DISA灌注代谢显像及门控心肌灌注显像。结果:36例DISA显像中,26例为灌注/代谢不匹配型,提示心肌存活,10例为灌注代谢匹配型,提示心肌梗死,无存活心肌,心功能正常患者30例。结论:一日法DISA心肌灌注/代谢显像与门控法心肌灌注显像,可全面评价糖尿病心肌病患者的心肌灌注、存活及心功能状况。     

Linkage with cathepsin B-sensitive dipeptide promotes the <Emphasis Type="Italic">in vitro</Emphasis> and <Emphasis Type="Italic">in vivo</Emphasis> anticancer activity of PEGylated tumor necrosis factor-alpha (TNF-α) against murine fibrosarcoma

To improve the pharmacological profile of tumor necrosis factor alpha (TNF-α), we have synthesized a new PEGylated prodrug, PEG-vcTNF-α, using a cathepsin B-sensitive dipeptide (valine-citrulline, vc) to link branched PEG and TNF-α. PEG-modified TNF-α without the dipeptide linker (PEG-TNF-α) and unconjugated TNF-α were also tested as controls. It was found for the first time that TNF-α released from PEG-vcTNF-α was specifically dependent on the presence of cathepsin B. PEG-vcTNF-α induced higher cytotoxicity and greater apoptosis against L929 murine fibrosarcoma cells than PEG-TNF-α. Reversal of these effects by a cathepsin-B inhibitor confirmed that these effects were mediated by cathepsin B-specific release of TNF-α. In vivo pharmacokinetics studies demonstrated that the plasma stability of PEG-vcTNF-α was significantly increased compared to TNF-α. Finally, the improved anticancer efficacy of PEG-vcTNF-α and the distinct activities among the three formulations confirmed the positive contribution of both PEGylation and the dipeptide linkage to the improved drug-like properties of PEG-vcTNF-α. The results here indicate that linking proteins and PEG via the cathepsin B-sensitive dipeptide may be a promising strategy for developing protein therapeutics.