Purification of plasmid DNA vectors by aqueous two-phase extraction and hydrophobic interaction chromatography

The current study explores the possibility of using a polyethyleneglycol(PEG)-ammonium sulphate aqueous two-phase system (ATPS) as an early step in a process for the purification of a model 6.1 kbp plasmid DNA (pDNA) vector. Neutralised alkaline lysates were fed directly to ATPS. Conditions were selected to direct pDNA towards the salt-rich bottom phase, so that this stream could be subsequently processed by hydrophobic interaction chromatography (HIC). Screening of the best conditions for ATPS extraction was performed using three PEG molecular weights (300, 400 and 600) and varying the tie-line length, phase volume ratio and lysate load. For a 20% (w/w) lysate load, the best results were obtained with PEG 600 using the shortest tie-line (38.16%, w/w). By further manipulating the system composition along this tie-line in order to obtain a top/bottom phase volume ratio of 9.3 (35%, w/w PEG 600, 6%, w/w NH4)2 SO4), it was possible to recover 100% of pDNA in the bottom phase with a three-fold increase in concentration. Further increase in the lysate load up to 40% (w/w) with this system resulted in a eight-fold increase in pDNA concentration, but with a yield loss of 15%. The ATPS extraction was integrated with HIC and the overall process compared with a previously defined process that uses sequential precipitations with iso-propanol and ammonium sulphate prior to HIC. Although the final yield is lower in the ATPS-based process the purity grade of the final pDNA product is higher. This shows that it is possible to substitute the time-consuming two-step precipitation procedure by a simple ATPS extraction.     

Interactions of dicarboxylic porphyrins with membranes in relation to their ionization state

The interactions of dicarboxylic porphyrins with membrane systems are discussed with particular emphasis on the effect of the charge of the porphyrin and the nature of the side-chains. The incorporation of hematoporphyrin or related dicarboxylic porphyrins within small unilamellar vesicles as membrane models is favored by a decrease of the pH in the range of physiological pH values. This effect might play an important role in the retention of porphyrins by tumors, which are more acidic than normal tissues. Kinetics studies also show that the partition of the porphyrin between the lipidic bilayer and the aqueous phase is governed by its release rate rather than by its incorporation rate.     

5-Aminolaevulinic acid-mediated photodynamic therapy in multidrug resistant leukemia cells

To verify if photodynamic therapy (PDT) could overcome multidrug resistance (MDR) when it it applied to eradicate minimal residual disease in patients with leukemia, we investigated the fluorescence kinetics of 5-aminolaevulinic acid (ALA)-induced protoporphyrin IX (PpIX) and the effect of subsequent photodynamic therapy on MDR leukemia cells, which express P-glycoprotein (P-gp), as well as on their parent cells. Evaluation of PpIX accumulation by flow cytometry showed that PpIX accumulated at higher levels in mdr-1 gene-transduced MDR cells (NB4/MDR) and at lower levels in doxorubicin-induced MDR cells (NOMO-1/ADR) than in their parent cells. A P-gp inhibitor could not increase PpIX accumulation. Measurement of extracellular PpIX concentration by fluorescence spectrometry showed that P-gp did not mediate the fluorescence kinetics of ALA-induced PpIX production. Assessment of ferrochelatase activity using high-performance liquid chromatography indicated that PpIX accumulation in drug-induced MDR cells was probably regulated by this enzyme. Assessment of phototoxicity of PDT using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay showed that PDT was effective in NB4, NB4/MDR, NOMO-1 and NOMO-1/ADR cells, which accumulated high levels of PpIX, but not effective in K562 and K562/ADR cell lines, which accumulated relatively low levels of PpIX. These findings demonstrate that P-gp does not mediate the ALA-fluorescence kinetics, and multidrug resistant leukemia cells do not have cross-resistance to ALA-PDT.     

脱臭蒜素胶囊的研制

介绍了大蒜的主要化学成分及食疗价值,探讨了大蒜的脱臭机理,阐述了脱臭蒜素胶囊的制造技术.     

Recent progress in polymer-based gene delivery vectors

The gene delivery system is one of the three components of a gene medicine, which is the bottle neck of current gene therapy. Nonviral vectors offer advantages over the viral system of safety, ease of manufacturing, etc. As important nonviral vectors, polymer gene delivery systems have gained increasing attention and have begun to show increasing promising. In this review, the fundamental and recent progress of polymer-based gene delivery vectors is reviewed.     

多曲方丝弓矫治技术的三维有限元分析

采用三维有限元法对后倾弯多曲方丝弓在四类牵引力作用下的矫治效果进行分析，在此基础上研究多曲方丝弓的整体力学特性，比较施加不同牵引力矫治的生物力学效果，揭示多曲方丝弓技术矫治特点。结果表明，无牵引力时，以第一、二前磨牙之间为界，多曲方丝弓矫治分别使前后牙向前后倾斜，且对前牙有压低作用；长Ⅲ类牵引时，对后牙的后倾作用有所增大，且传递力的性能比长Ⅱ类牵引好；Ⅱ类牵引力作用下的MEAW弓丝矫治对前牙有伸长作用，短Ⅱ类牵引对磨牙的伸长作用和对前牙的作用效果均小于长Ⅱ类牵引。因此，多曲方丝弓矫治技术有利于下颌Spee氏曲线的整平和牙齿的个别调控，对牵引力的传递作用强，同时对开拾错铪矫治有很好的效果。     

跨栏跑教学中学生恐惧心理克服实验研究

采用统计法、实验法等分析方法,对我院随机抽样学生进行跨栏跑教学实验,认为学生对跨栏跑教学普遍存在有一定的恐惧心理.本文根据跨栏跑教学的运动规律采取了相关训练方法进行研究.结果表明,运用此方法是切实可行、行之有效的.     

朊病毒和朊病毒疾病治疗的研究进展

介绍了朊病毒的基本特征、化学结构以及可能具有的生理功能：参与神经系统功能的维持；通过抗氧化途径而保护神经系统细胞免受氧化损伤；参与淋巴细胞的信号转导；参与核酸代谢，给出了朊病毒治疗的几种方法，并分析了这些方法的利弊，提出了朊病毒研究拟待解决的9个问题。     

永磁体的开路剩磁及其临床意义

为了得到永磁体开路剩磁的空间变化规律。本文通过实验测量后,经统计学处理,得出了永磁体开路剩磁按负指数规律衰减的结论,从而为临床上的磁场疗法提供了理论依据。     

顺铂联合白细胞介素-2治疗恶性胸腔积液32例

目的：探讨胸腔注入顺铂（DDP）联合白细胞介素-2（IL-2）治疗恶性胸腔积液疗效。方法：58例患者均病理确诊为恶性胸腔积液，随机分为二组，治疗组（32例）在胸腔内注入DDP，白细胞介素-2，对照组（26例）在胸腔内注A．DDP，两组均在1周及2周后各重复注药1次，共3次，观察疗效，毒副反应及生活质量。结果：治疗组总有效率为84．4％，对照组57．7％，差异有显著性（P〈0．015）。治疗组Kamofsky评分〉70分者较对照组有显著提高（P〈0．05），两组在毒性反应上无显著性差异（P〉0．05）。结论：顺铂白介素-2胸腔灌注能有效控制恶性胸腔积液，毒副作用少，联合疗效肯定。