新工科背景下生物反应工程课程的多元化教学模式 改革与探索

生物反应工程作为一门理论性与应用性都很强的专业课程,在生物工程等相关专业的课程设置中处于桥梁和纽带地位,对新型应用型工科人才的培养发挥着重要的作用。但由于该课程中公式等抽象理论知识过多,导致学生学习效率十分低下。因此,为了适应新工科教育背景下对创新型人才培养的需求,提高学生的学习兴趣和积极性,并培养学生的自主学习等创新能力,教学团队在课程教学中通过引入虚拟仿真技术、开展微课教学、采用案例式教学模式、利用科研平台等多元化方式,对该课程的教学模式、方法和手段尝试改革和探索,取得了一定的教学效果,并就此进行了一些探讨,以期能为相关课程的教学改革提供一些思路和启示。     

利用CRISPR-Cas9技术失活黑曲霉中果胶酶基因及突变株性能评价*

我国果胶酶制剂使用广泛但专一性不高,高效、专一的果胶酶制剂在市场上仍然匮乏。利用基因工程技术改造果胶酶生产菌株——黑曲霉来生产单一成分的果胶酶成为解决果胶酶应用需求的一种有效方案。构建一种高效的CRISPR-Cas9基因编辑技术,可为构建高产单一性果胶酶的黑曲霉底盘菌株提供有效的基因编辑工具。首先敲除产果胶酶黑曲霉基因组上的pyrG基因构建尿嘧啶营养缺陷型菌株AnΔpyrG,并在AnΔpyrG菌株的pyrG基因位点定点整合Cas9基因表达盒和pyrG基因表达盒,构建组成型表达Cas9基因的黑曲霉菌株AnCas9,再构建含有gpdA启动子、锤头结构核酶、HDV核酶的稳定性表达sgRNA的pLM2-sgRNA质粒,建立CRISPR-Cas9基因编辑体系。利用该技术失活AnCas9菌株中的2个聚半乳糖醛酸酶基因4978020和4983861来检测构建的CRISPR-Cas9基因编辑效率并检测4978020基因功能缺失菌株的表型变化和产酶变化,结果表明果胶酶基因编辑效率大于50%,AnΔ4978020的表型和果胶酶酶活性与出发菌株均无明显变化。在黑曲霉中成功构建了高效的Cas9基因编辑技术,4978020基因功能缺失也不影响菌株表型,为构建高产单一性果胶酶黑曲霉底盘菌株奠定基础。     

奈妥吡坦/β-环糊精包合物的制备、评价与表征

目的:制备奈妥吡坦/β-环糊精包合物,用以提高奈妥吡坦的水溶性。方法:采用饱和水溶液法,制备奈妥吡坦/β-环糊精包合物;以载药量为指标,考察奈妥吡坦与β-环糊精的质量比(芯壁比)、包合温度、包合时间、搅拌速度的影响。基于单因素试验结果,采用正交设计实验对制备处方和工艺进行优化,得到最优奈妥吡坦/β-环糊精包合物,并对其包封率、载药量及稳定性进行体外评价。采用傅里叶红外光谱和X射线粉末衍射分析法,对最优奈妥吡坦/β-环糊精包合物进行表征。结果:奈妥吡坦/β-环糊精包合物的最佳优制备条件为:芯壁比1:20,包合温度为40℃,包合时间为20 min、搅拌速度为150 r/min。该条件制备制得的奈妥吡坦/β-环糊精包合物的载药量为4.73%,包合率为94.8%且在水溶液中的稳定性良好。傅里叶红外光谱和X射线衍射结果表明奈妥吡坦/β-环糊精包合物的成功制备。结论:成功制备了奈妥吡坦/β-环糊精包合物,奈妥吡坦的水溶性提高了1500倍,为奈妥吡坦水溶性新制剂的研发提供了实验基础。     

呼吸中的疾病诊断标志物及检测技术

人类呼出气体中的各种化合物能提供各种疾病和健康状况的重要信息。近年来,由于红外、电化学、化学发光等新技术的重大突破和质谱仪的使用,使得在极低浓度下精确测量呼出的挥发性有机化合物(VOCs)和气溶胶颗粒成为可能,呼吸检测领域因而取得了重大进展:,呼吸检测因其可以作为一种实时、快速和无创的方法来评估和监测各种疾病与健康状况信息,在科学研究、临床运用中引起了广泛关注。本综述主要概述呼出气体成分分析方法及在疾病诊断中的研究与应用情况,旨在为将来疾病的实时、快速和无创诊断提供一种新的策略.     

Determination of protein concentration in downstream biomanufacturing processes by in-line index of refraction

Protein concentration determination is a necessary in-process control for the downstream operations within biomanufacturing. As production transitions from batch mode to an integrated continuous bioprocess paradigm, there is a growing need to move protein concentration quantitation from off-line to in-line analysis. One solution to fulfill this process analytical technology need is an in-line index of refraction (IoR) sensor to measure protein concentration in real time. Here the performance of an IoR sensor is evaluated through a series of experiments to assess linear response, buffer matrix effects, dynamic range, sensor-to-sensor variability, and the limits of detection and quantitation. The performance of the sensor was also tested in two bioprocessing scenarios, ultrafiltration and capture chromatography. The implementation of this in-line IoR sensor for real-time protein concentration analysis and monitoring has the potential to improve continuous bioprocess manufacturing.     

微小RNA分析技术研究进展

微小RNA（miRNA）是一类起重要调控作用的非编码小分子RNA。准确分析组织或细胞中miRNA的表达水平是研究其生物学功能的基础。近年,研究者开发出多种方法检测不同生理、病理过程中miRNA的差异表达,发现miRNA的异常表达与癌症等多种疾病密切相关。目前,miRNA已逐渐成为重要的疾病诊断生物标志物乃至治疗靶点。miRNA的分析技术贯穿miRNA的研究和药物研发过程,并起到关键作用。我们针对miRNA的不同研究阶段所采用的定性和定量分析方法,着重阐述了用于初始miRNA研究的克隆测序类技术、分析miRNA表达谱的高通量芯片技术、研究具体目标miRNA及其前体表达的qPCR和改良Northern印迹技术,以及将修饰后miRNA作为药物的药代动力学评价技术。     

Forest ecosystem inventory and monitoring as a framework for terrestrial natural renewable resource survey programmes

ABSTRACT

The established practice of forest ecosystem inventory and monitoring is recognised as a main support for terrestrial natural renewable resource survey programmes. Inventory and monitoring programmes focused on an overall assessment of ecosystem attributes evolving into global environmental survey programmes have been devised, but implementation is still quite contradictory. The state-of-the-art is discussed here, with special reference to the European Union and Italy. Topical issues are reviewed, with selective concern to: remote sensing capability, probability sampling, forest type (habitat) classification and landscape ecology, sustainable management indicators. Benefits brought by information technology are highlighted. Its development and the implementation of approaches based on a sound “per habitat” landscape ecological perspective will bring unique benefits, thus leading to an effective integration among sector surveys aimed at global environmental inventory/monitoring.     

Photoluminescence of Alq3‐ and Tb‐activated aluminium–tris(8‐hydroxyquinoline) complex for blue chip‐excited OLEDs

The tris(8‐hydroxyquinoline)–aluminium complex is the most important and widely studied as electron transporting and green light emitting material. Alq₃ and Tb_xAl_(1‐x)q₃ have been synthesized (where x = 0.1, 0.3, 0.5, 0.7 and 0.9) and blended films of Alq₃ and Tb_xAl_(1‐x)q₃ with PMMA and PS at different percentage weight (wt%) concentrations (e.g., 0.1, 1, 5, 10, 25 and 50 wt%) have been prepared. The synthesized materials and their blended thin films have been characterized by a photoluminescence (PL) technique; the synthesis and PL characterization are reported in this paper. The synthesized metal complex shows bright emission of green light with blue light excitation (440 nm) and the prepared Tb_xAl_(1‐x)q₃ phosphor may be applicable in blue chip‐excited OLEDs for the newly developed wallpaper lighting technology. Copyright © 2012 John Wiley & Sons, Ltd.     

Integrating frugivory and animal movement: a review of the evidence and implications for scaling seed dispersal

General principles about the consequences of seed dispersal by animals for the structure and dynamics of plant populations and communities remain elusive. This is in part because seed deposition patterns emerge from interactions between frugivore behaviour and the distribution of food resources, both of which can vary over space and time. Here we advocate a frugivore‐centred, process‐based, synthetic approach to seed dispersal research that integrates seed dispersal ecology and animal movement across multiple spatio‐temporal scales. To guide this synthesis, we survey existing literature using paradigms from seed dispersal and animal movement. Specifically, studies are discussed with respect to five criteria: selection of focal organisms (animal or plant); measurement of animal movement; characterization of seed shadow; animal, plant and environmental factors included in the study; and scales of the study. Most studies focused on either frugivores or plants and characterized seed shadows directly by combining gut retention time with animal movement data or indirectly by conducting maternity analysis of seeds. Although organismal traits and environmental factors were often measured, they were seldom used to characterize seed shadows. Multi‐scale analyses were rare, with seed shadows mostly characterized at fine spatial scales, over single fruiting seasons, and for individual dispersers. Novel animal‐ and seed‐tracking technologies, remote environmental monitoring tools, and advances in analytical methods can enable effective implementation of a hierarchical mechanistic approach to the study of seed dispersal. This kind of mechanistic approach will provide novel insights regarding the complex interplay between the factors that modulate animal behaviour and subsequently influence seed dispersal patterns across spatial and temporal scales.