A review of near infrared spectroscopy and chemometrics in pharmaceutical technologies

Near-infrared spectroscopy (NIRS) is a fast and non-destructive analytical method. Associated with chemometrics, it becomes a powerful tool for the pharmaceutical industry. Indeed, NIRS is suitable for analysis of solid, liquid and biotechnological pharmaceutical forms. Moreover, NIRS can be implemented during pharmaceutical development, in production for process monitoring or in quality control laboratories.This review focuses on chemometric techniques and pharmaceutical NIRS applications. The following topics are covered: qualitative analyses, quantitative methods and on-line applications. Theoretical and practical aspects are described with pharmaceutical examples of NIRS applications.     

介入途径下中药白芨提取物作为基因递送载体的可行性

目的:探讨经介入途径中药白芨提取物作为基因递送载体的可行性.方法:从中药白芨中提取其有效成分-白芨多糖, 采用胺化还原法制备阳离子型白芨多糖,体外实验检测该阳离子型多糖对质粒DNA的结合与保护作用, 以及阳离子白芨多糖载基因复合物对肝癌细胞系HepG2的转染, 进一步通过介入途径经肝动脉给予该复合物, 以GFP作为报告基因检测复合物在体内对活体兔肝细胞的转染.结果:所制备的阳离子型白芨多糖载基因复合物可以结合并保护质粒DNA免受DNA酶的降解; 体外实验证实该复合物可以转染入体外培养肝癌细胞, 以阳离子白芨多糖作为转染载体时转染效率要低于脂质体组, 差异具有统计学意义(28.87%±3.27% vs 36.64%±6.87%,P <0.05). 采用介入方法经肝动脉给药时复合物能靶向转染入活体兔肝细胞内并实现表达.结论:采用介入途径经肝动脉给药时阳离子白芨多糖载基因复合物可以实现活体肝细胞靶向转染, 有望作为一种新型的多聚阳离子型的基因载体在基因治疗中发挥作用.     

Boundary sequences stabilize transgene expression from subtle position effects in retroviral vectors

Transgene expression shut-down, attenuation and/or variability from integrated retroviral vectors pose a major obstacle to gene therapy trials involving hematopoietic cells. We have undertaken a systematic assessment of the behavior of different configurations containing IFN-β SAR and/or 5′HS4 β-globin insulator sequences within a gammaretroviral vector optimized for high-level expression, focusing on the long-term achievement of stable, homogeneous transgene expression in the successfully transduced cells. Introduction of these cis regulatory elements did not perturb virus production and stability. Conversely, the SAR/5′HS4 insulator combination appeared to increase the homogeneity of EGFP expression in mass cultures. Furthermore, a clonal analysis of the dispersion of EGFP expression revealed that the IFN-SAR/5′HS4 insulator dyad was particularly effective in reducing the variability of transgene expression when both sequences were placed in opposite orientations within the retroviral backbone. These results may prove useful for the design of more stable retroviral expression cassettes able to counteract chromosomal position effects.     

Shifting priorities in vector biology to improve control of vector-borne disease

Vector control remains the primary measure available to prevent pathogen transmission for the most devastating vector-borne diseases (VBDs): malaria, dengue, trypanosomiasis, filariasis, leishmaniasis, and Chagas disease. Current control strategies, however, are proving insufficient and the remarkable advances in the molecular biology of disease vectors over the last two decades have yet to result in tangible tools that effectively reduce VBD incidence. Here we argue that vector biologists must fundamentally shift their approach to VBD research. We propose an agenda highlighting the most critical avenues to improve the effectiveness of vector control. Research priorities must be diversified to support simultaneous development of multiple, alternative control strategies. Knowledge across relevant diseases and disciplines should be better integrated and disease prevention efforts extended beyond the academic sector to involve private industry, ministries of health, and local communities. To obtain information of more immediate significance to public health, the research focus must shift from laboratory models to natural pathogen-transmission systems. Identification and characterization of heterogeneities inherent to VBD systems should be prioritised to allow development of local, adaptive control strategies that efficiently make use of limited resources. Importantly, increased involvement of disease-endemic country (DEC) scientists, institutes, and communities will be key to enhance and sustain the fight against VBD.     

pEGFP-hApelin-R重组真核表达载体的构建及在HEK293细胞中的表达

目的构建与增强型绿色荧光蛋白（enhanced green fluorescent protein,EGEP）融合的人apelin受体（Apelin receptor,apelin-R）真核表达载体。方法以质粒pcDNA3.1-hApelin-R为模板,PCR方法扩增人apelin受体。扩增的人apelin受体用EcoRⅠ和BamHⅠ双酶切,同时用这两种酶双酶切质粒peGFP-C1。然后将两种酶切产物按常规方法连接、转化大肠杆菌Top10。挑取菌落培养,提取质粒,然后进行酶切鉴定,最后进行测序。将测序正确的重组载体用脂质体法转染人胚胎肾（human embryonic kidney293,HEK293）细胞,共聚焦显微镜观察。提取转染细胞的总蛋白,进行Westernblot检测。结果扩增出一条约1200bp的片段,与预期的apelin受体大小相符。酶切结果显示,重组质粒pEGFP-hApelin-R被切成两条片段,其中一条为peGFP-C1载体大小,另一条为目的片段大小。经测序鉴定,序列与GenBank（NM_005161）中的序列高度同源。共聚焦显微镜观察显示,人apelin受体主要在细胞膜上表达。Westernblot结果显示在相对分子质量69000处有一蛋白条带,与预期大小相符。结论构建成功pEGFP-hApelin-R重组表达载体,此表达载体可用于检测apelin受体和κ型阿片受体（kappa opioid receptor,KOR）或与其他受体间的相互作用。     

Viral Hybrid Vectors for Somatic Integration - Are They the Better Solution?

The turbulent history of clinical trials in viral gene therapy has taught us important lessons about vector design and safety issues. Much effort was spent on analyzing genotoxicity after somatic integration of therapeutic DNA into the host genome. Based on these findings major improvements in vector design including the development of viral hybrid vectors for somatic integration have been achieved. This review provides a state-of-the-art overview of available hybrid vectors utilizing viruses for high transduction efficiencies in concert with various integration machineries for random and targeted integration patterns. It discusses advantages but also limitations of each vector system.     

VFM评价正常成人左室心腔血流流场运动状态的初步研究

目的:运用VFM( Vector Flow Mapping)成像探讨正常成人左心室腔内收缩期血流流场特征.方法:健康成人志愿者40例(男22例,女18例),二维超声取心尖三腔观左室腔彩色血流信息动态图像存贮并脱机分析,将取样线分别置于左室腔基底段、中尖段和心尖段,获取心动周期内经取样线部位左室腔血流时间速度积分(Time-Flow curve, TF)变化曲线,记录基底段、中间段及心尖段收缩早期速度时间积分(ES-TF)、收缩中期速度时间积分(MS-TF)、收缩晚期速度时间积分(LS-TF).观察收缩期血流流场特征,观察涡流出现时间,测量涡流直径(横径Dx,)及涡流的最大向量速度(Vmax).结果:正常人左室心腔血流流场呈规则变化,收缩早期二尖瓣前叶下方可见涡流显示,收缩中期及晚期涡流消失.收缩早、中、晚期,血流时间速度积分测值由心尖段经中间段至基底段逐渐递增,差异均有统计学意义(P<0.05).收缩早期涡流直径约为(25.74±7.32)mm, 涡流最大向量速度均背离探头,大小为(38.31±14.25)cm/s,朝向探头速度大小为(11.42±6.28) cm/s.结论:应用VFM技术可清晰显示左室心腔内血流流场状态,左室腔血流速度积分测值可直观反应心腔内血流动力学变化.     

2000～2007年怀远县疟疾流行特征及媒介监测结果分析

怀远县2000～2007年全县疫情报告系统报告疟疾病例数分别为49、481、1905、2464、1321、1218、1167和625例;疟疾发病主要集中在5～11月份。2005～2007年疟疾病人及时抗疟治疗率仅为2.97%,复发率为2.61%;2002～2007年血检发热病人140641人次,阳性率为3.85%;2003～2007年人群疟疾抗体检测6075人次,阳性率为6.01%。怀远县传疟媒介为单一中华按蚊。     

超声量化技术在大鼠脂肪肝模型中应用的初步研究

目的：探讨超声量化技术评价大鼠肝脏脂肪变的价值。方法：大鼠ig酒精高脂乳液12周,制备脂肪肝模型。测量正常大鼠及模型大鼠肝脏二维声像图的灰度值,检测肝组织三酰甘油（TG）、总胆固醇（TC）,并将肝脏组织做病理切片,观察肝细胞脂变情况。结果：与正常组比较,模型组超声图像近区、远区灰度显著增加,肝脏TG、TC明显升高。组织病理学分析可见模型大鼠肝脏典型脂肪样变。结论：超声量化技术可作为一种无创性量化方法用于判断大鼠肝脏脂肪变化。