环境微生物学翻转课堂教学改革探索

南京大学在环境微生物学教学中大力推进翻转课堂教学改革与实践。教学内容上注重夯实微生物基础,与实际生活和环境问题结合,重新编排环境微生物学课程知识体系,同时进行应用型和研究型的差异化教学。教学模式上,将启发式、互动式和研究型教学引入课堂,同时运用逐步推进式翻转课堂教学模式,注重反馈式教学,以学为本,以提高学生对环境微生物学的理解和应用。     

Transmembrane H + and Ca 2+ fluxes through extraradical hyphae of arbuscular mycorrhizal fungi in response to drought stress

丛枝菌根真菌(AMF)能够和大多数陆地植物形成共生体系, 对于植物生长发育和适应各种逆境胁迫具有重要作用。很多研究表明干旱胁迫下AMF能够促进宿主植物对水分的吸收从而增强植物抗旱能力, 但目前针对AMF根外菌丝响应水分胁迫的生理变化以及AMF与宿主植物逆境信号交流的研究并不多。该研究利用AMF Rhizophagus irregularis和胡萝卜(Daucus carota var. sativa)毛状根双重无菌培养体系获得纯净根外菌丝, 向培养基添加聚乙二醇(PEG)模拟干旱胁迫, 运用场发射扫描电子显微镜(FE-SEM-EDS)观察干旱胁迫对AMF根外菌丝形态的影响, 同时采用非损伤微测技术(NMT)观测根外菌丝跨膜H ⁺和Ca ²⁺离子流变化。结果发现, PEG处理1 h后菌丝尖端和侧面发生H ⁺外流和强烈的Ca ²⁺内流, 荧光探针分析也显示菌丝胞内pH值显著上升、Ca ²⁺浓度增加; PEG处理24 h后菌丝形态发生明显变化, 培养基pH值降低, P、Ca、Fe等元素在菌丝际积累。这些试验结果表明, 干旱胁迫下AMF根外菌丝跨膜H ⁺和Ca ²⁺流发生变化, 促进了菌丝与环境之间的物质交换。菌丝酸化生长环境有利于养分吸收, 并促进AMF与宿主植物之间的信号交流以增强植物的耐旱性。     

Long term effects of ART: What do animals tell us?

Early stages of mammalian embryonic development are now known to be very sensitive to their microenvironment, with long term effects on fetal, postnatal, and adult health, thus extending to these early stages the concept of Developmental Origin of Health and Disease (DoHaD). In this scientific context, and with 3% of births in developed countries, safety of Assisted Reproductive Techniques procedures becomes a matter of concern. Besides, embryo technologies in domestic mammals, using huge number of embryos, do not seem to evidence heavy impacts on adult phenotypes. This paper first discusses what can or cannot be concluded from farm animal data, then develops long term effects of ART procedures (ovarian stimulation, in vitro fertilization and embryo culture) evidenced in model species (mainly mouse model). Recent literature demonstrates both individual and cumulative effects of each ART procedure on fetal and postnatal phenotypes. In a second part, because they are sources for further perturbations, immediate effects of ART on early embryo phenotypes at the cellular and molecular levels are described in both farm animals and model species. Mechanistic hypotheses supporting these ART induced phenotypic alterations are subsequently considered. Finally, taking into account interspecies differences in the mechanisms likely to be involved, the relevance of results obtained in animal models for human ART are discussed.     

玉米酵母双杂交cDNA文库的构建及ZmCEN互作蛋白的筛选

为阐明玉米中心蛋白(ZmCEN)的生物学功能,采用酵母双杂交技术对其互作蛋白进行研究。提取玉米(Zea mays L.)自交系‘郑58’幼苗的总RNA,利用SMART技术反转录合成ds cDNA,构建以pGBKT7为载体的酵母双杂交cDNA文库;依据ZmCEN基因的CDS序列设计引物,构建重组诱饵载体(pGBKT7-ZmCEN)转化酵母菌株Y2HGold,检测诱饵载体的毒性与自激活能力后,筛选与玉米中心蛋白(ZmCEN)互作的猎物蛋白。将筛选的互作蛋白NAC67和TONNEAU1b(TON1b)再次验证相互作用,并选取互作蛋白TON1b,采用BiFC实验分别构建ZmCEN-pSPYNE和TON1b-pSPYCE BiFC半分子重组载体,转化拟南芥原生质体,进一步验证它们在细胞内的互作;并利用Uniprot和KEGG在线网站对互作蛋白进行gene ontology(GO)注释分析。结果表明:玉米全株幼苗的cDNA文库库容量达到2.56×107 CFU,文库滴度5.36×108 CFU/mL,符合建库要求。经检测诱饵载体无毒性也无自激活功能,所筛选的cDNA文库经测序和Blast比对分析以及共转验证,最终得到28个与诱饵蛋白ZmCEN互作的蛋白质。GO注释显示互作蛋白参与的生物过程有21种。BiFC结果显示,蛋白TON1b与ZmCEN在拟南芥原生质体细胞内互作而形成互补,从而产生黄色荧光,进一步证实了两者存在互作关系。酵母双杂交系统cDNA文库的成功构建与筛选,为进一步研究玉米ZmCEN及其与互作蛋白的作用机制奠定了基础。     

植物非整倍体研究进展

非整倍体(aneuploid)是指相对于正常个体(euploid)的染色体组增加、减少一条或若干条染色体的生物个体。由于非整倍体个体存在基因剂量效应的不平衡性(gene-dosage imbalance),非整倍体个体往往会表现严重的表型缺陷(aneuploid syndrom),如发育迟缓,个体矮小,难以繁殖后代等。在人类中,最为典型的例子为导致新生儿智力缺陷的唐氏综合症,由额外的一个21号染色体拷贝(部分拷贝)引起。此外,大多数癌细胞类型表型为严重的非整倍体。在大多情况下,非整倍体对于动物及人类是致命的,而植物对于非整倍体则往往表现出较强的耐受力,特别是在异源多倍体植物中。植物非整倍体对于植物的遗传、育种研究有重要意义,在基因及分子标记的物理位置确定,基因转移,连锁群与染色体的对应关系的确立上具有无可比拟的优势。该文综述了近些年来有关植物非整倍体研究的结果,介绍了非整倍体的几种重要成因和有关非整倍体鉴定手段的变迁,阐述了植物非整倍体对个体表型、基因表达以及表观遗传方面的影响,重点讨论了非整倍体在植物进化、基因组序列测定以及遗传改良方面的潜在作用。同时,探讨了植物非整倍体研究的新思路,以及利用非整倍体促进相关植物遗传改良、育种研究的新方法。     

Construction Materials from Stainless Steel Slags: Technical Aspects,Environmental Benefits,and Economic Opportunities

State‐of‐the‐art technologies that implement theindustrial ecology concept only make it to the market if environmental gains and economic benefits are significant. Therefore, the article investigates, in an interdisciplinary way, two innovative technologies that valorize stainless steel (SS) slags as block masonry (bricks): carbonation and thermo‐alkali‐activation. The technical, environmental, and economic features of three SS bricks—solid bricks, perforated bricks, and lightweight aerated blocks—are compared to commercially available construction materials. Although the produced bricks meet industrial standards, technical challenges, such as optimization of alkali addition and use of metal molds, should be dealt with before upscaling to industrial production. A cradle‐to‐gate life cycle assessment that aggregates the results of the various impact categories shows that the environmental impact of solid and perforated SS bricks is lower than the impact of conventional clay‐baked bricks owing to the avoidance of additives for slag stabilization and energy consumption for sintering clay. The impact of aerated SS bricks was found to be similar to the commercially available aerated blocks. More specifically, the carbon dioxide uptake from carbonation reduces the overall environmental impact, whereas use of alkalis increases the impact. A strengths weaknesses opportunity threats analysis highlights the economic advantages of SS bricks originating from lower energy requirements, reduced dependence on primary resources, and improved metal recovery from slag. However, in order to apply the innovative technologies at industrial scale, challenges related to processing conditions, feedstock variability, and potential competition from existing brick suppliers have to be overcome.     

传统开腹手术与腹腔镜手术治疗子宫肌瘤的效果分析

目的探讨传统开腹手术与腹腔镜手术治疗子宫肌瘤的临床疗效。方法选择2013年2月~2015年11月在我院接受治疗的子宫肌瘤患者140例为研究对象,按患者自愿原则分为对照组和观察组各70例,对照组采用传统开腹手术治疗,观察组采用腹腔镜手术治疗,并对两组患者手术时间、术中出血量、术后感染情况和住院时间进行比较。结果观察组在手术时间、术中出血量、术后感染率及住院时间等方面均优于对照组,差异具有显著统计学意义(P0.05)。结论腹腔镜下子宫肌瘤切除术与传统开腹手术比较,具有出血量少、住院时间短、恢复快、并发症少等优点,值得临床推广应用。     

基于组织工程研究的可降解支架材料选择策略

目前,器官或组织移植是治疗器官衰竭或大范围组织缺损唯一长期有效的方法,但存在供体短缺、免疫排斥等问题。组织工程技术作为一种潜在的替代治疗方法,支架材料的选择是其中具有决定意义的组成部分。组织工程支架材料按其来源可分为天然及其改性修饰材料、人工合成与复合支架材料3种。组织工程目的就是修复临床上的病损组织或器官,并达到较理想的结构和功能的恢复。因此组织工程支架也必须从基本性质上具有一定的仿生化结构及功能,即"活"支架,这样才能彻底代替病损组织或器官。通过多种支架材料的优化组合(即材料的复合),对材料进行表面改性、制备工艺优化及添加细胞因子缓释微球等技术,模拟病损器官组织的特性及周围环境,有望打开组织工程的新局面。理想的组织工程支架应当以临床需要为根本目的,依靠材料学、分子生物学、工程学等多学科间的交叉研究,取各家之长,优化配比组合,达到仿生的目的。本课题组前期工作已经将骨髓间充质干细胞体外诱导分化为胆管上皮样细胞,并设计出左旋聚乳酸/聚己内酯共聚物(PLCL)胆道支架,内部混有包含生长因子的纳米缓释微球,供细胞因子的远期释放,支架内表面涂有基质胶/胶原混合层,且胶内加入bFGF、EGF,提供诱导因子的早期释放。将诱导细胞与PLCL胆道支架复合,制备组织工程胆管。文中综述了现存各类支架材料的研究状况,简单介绍了制备工艺、表面修饰等影响支架性能的因素,力求探索组织工程支架材料的选择策略。     

正交试验优选艾叶总黄酮索氏提取工艺

采用正交试验法,对艾叶总黄酮索氏提取工艺进行考察,确定艾叶总黄酮最优提取工艺。结果发现,以总黄酮得率作为考察指标,艾叶中总黄酮的最佳提取条件为:料液比为1∶35,浸提温度为80℃,浸提时间为3h,乙醇体积分数为70%,艾叶总黄酮得率高达3.85%。     

Production of greenhouse‐grown biocrust mosses and associated cyanobacteria to rehabilitate dryland soil function

Mosses are an often‐overlooked component of dryland ecosystems, yet they are common members of biological soil crust communities (biocrusts) and provide key ecosystem services, including soil stabilization, water retention, carbon fixation, and housing of N₂ fixing cyanobacteria. Mosses are able to survive long dry periods, respond rapidly to precipitation, and reproduce vegetatively. With these qualities, dryland mosses have the potential to be an excellent dryland restoration material. Unfortunately, dryland mosses are often slow growing in nature, and ex situ cultivation methods are needed to enhance their utility. Our goal was to determine how to rapidly produce, vegetatively, Syntrichia caninervis and S. ruralis, common and abundant moss species in drylands of North America and elsewhere, in a greenhouse. We manipulated the length of hydration on a weekly schedule (5, 4, 3, or 2 days continuous hydration per week), crossed with fertilization (once at the beginning, monthly, biweekly, or not at all). Moss biomass increased sixfold for both species in 4 months, an increase that would require years under dryland field conditions. Both moss species preferred short hydration and monthly fertilizer. Remarkably, we also unintentionally cultured a variety of other important biocrust organisms, including cyanobacteria and lichens. In only 6 months, we produced functionally mature biocrusts, as evidenced by high productivity and ecosystem‐relevant levels of N₂ fixation. Our results suggest that biocrust mosses might be the ideal candidate for biocrust cultivation for restoration purposes. With optimization, these methods are the first step in developing a moss‐based biocrust rehabilitation technology.