Induction and Analysis of Epithelial to Mesenchymal Transition

Epithelial to mesenchymal transition (EMT) is essential for proper morphogenesis during development. Misregulation of this process has been implicated as a key event in fibrosis and the progression of carcinomas to a metastatic state. Understanding the processes that underlie EMT is imperative for the early diagnosis and clinical control of these disease states. Reliable induction of EMT in vitro is a useful tool for drug discovery as well as to identify common gene expression signatures for diagnostic purposes. Here we demonstrate a straightforward method for the induction of EMT in a variety of cell types. Methods for the analysis of cells pre- and post-EMT induction by immunocytochemistry are also included. Additionally, we demonstrate the effectiveness of this method through antibody-based array analysis and migration/invasion assays.     

Plant Life History and Residue Chemistry Influences Emissions of CO2 and N2O From Soil – Perspectives for Genetically Modified Cell Wall Mutants

Vascular plants have lignified tissues that transport water, minerals, and photosynthetic products throughout the plant. They are the dominant primary producers in terrestrial ecosystems and capture significant quantities of atmospheric carbon dioxide (CO₂) through photosynthesis. Some of the fixed CO₂ is respired by the plant directly, with additional CO₂ lost from rhizodeposits metabolized by root-associated soil microorganisms. Microbially-mediated mineralization of organic nitrogen (N) from plant byproducts (rhizodeposits, dead plant residues) followed by nitrification generates another greenhouse gas, nitrous oxide (N₂O). In anaerobic soils, reduction of nitrate by microbial denitrifiers also produces N₂O. The plant-microbial interactions that result in CO₂ and N₂O emissions from soil could be affected by genetic modification. Down-regulation of genes controlling lignin biosynthesis to achieve lower lignin concentration or a lower guaiacyl:syringyl (G:S) ratio in above-ground biomass is anticipated to produce forage crops with greater digestibility, improve short rotation woody crops for the wood-pulping industry and create second generation biofuel crops with low ligno-cellulosic content, but unharvested residues from such crops are expected to decompose quickly, potentially increasing CO₂ and N₂O emissions from soil. The objective of this review are the following: 1) to describe how plants influence CO₂ and N₂O emissions from soil during their life cycle; 2) to explain how plant residue chemistry affects its mineralization, contributing to CO₂ and N₂O emissions from soil; and 3) to show how modification of plant lignin biosynthesis could influence CO₂ and N₂O emissions from soil, based on experimental data from genetically modified cell wall mutants of Arabidopsis thaliana. Conceptual models of plants with modified lignin biosynthesis show how changes in phenology, morphology and biomass production alter the allocation of photosynthetic products and carbon (C) losses through rhizodeposition and respiration during their life cycle, and the chemical composition of plant residues. Feedbacks on the soil environment (mineral N concentration, soil moisture, microbial communities, aggregation) affecting CO₂ and N₂O emissions are described. Down-regulation of the Cinnamoyl CoA Reductase 1 (CCR1) gene is an excellent target for highly digestable forages and biofuel crops, but A. thaliana with this mutation has lower plant biomass and fertility, prolonged vegetative growth and plant residues that are more susceptible to biodegradation, leading to greater CO₂ and N₂O emissions from soil in the short term. The challenge in future crop breeding efforts will be to select tissue-specific genes for lignin biosynthesis that meet commercial demands without compromising soil CO₂ and N₂O emission goals.     

Effects of a transient oxic period on mineralization of organic matter to CH4 and CO2 in anoxic peat incubations

Rates of organic matter mineralization in peatlands, and hence production of the greenhouse gases CH₄ and CO₂, are highly dependent on the distribution of oxygen in the peat. Using laboratory incubations of peat, we investigated the sensitivity of the anoxic production of CH₄ and CO₂ to a transient oxic period of a few weeks’ duration. Production rates during 3 successive anoxic periods were compared with rates in samples incubated in the presence of oxygen during the second period. In surface peat (5–10‐cm depth), with an initially high level of CH₄ production, oxic conditions during period 2 did not result in a lower potential CH₄ production rate during period 3, although production was delayed ～1 week. In permanently anoxic, deep peat (50–55‐cm depth) with a comparatively low initial production of CH₄, oxic conditions during period 2 resulted in zero production of CH₄ during period 3. Thus, the methanogens in surface peal—but not in deep peat—remained viable after several weeks of oxic conditions. In contrast to CH₄ production, the oxic period had a negligible effect on anoxic CO₂ production during period 3, in surface as well as deep peat. In both surface and deep peat, CO₂ production was several times higher under oxic than under anoxic conditions. However, for the first 2 weeks of oxic conditions, CO₂ production in the deep peat was very low. Still, deep peat obviously contained facultative microorganisms that, after a relatively short period, were able to maintain a considerably higher rate of organic matter mineralization under oxic than under anoxic conditions.     

内蒙古不同类型草地土壤氮矿化及其温度敏感性

土壤氮矿化(Nitrogen mineralization)是土壤氮循环的重要环节,对土壤氮素供应以及植物生产力的维持具有十分重要的意义。沿中国东北草地样带(Northeastern China Transect, NECT)分别在典型草地、过渡草地及荒漠草地设置了3个实验样地,利用不同温度(5、10、15、20 ℃和25 ℃)和不同水分(30%、60%和90%土壤饱和含水量,Saturated soil moisture, SSM)的室内培养途径,探讨了不同类型草地的土壤氮矿化速率、土壤氮矿化的温度敏感性(Q₁₀)及其主要影响因素。实验结果表明:从典型草地至荒漠草地,土壤全碳、全氮、全磷、微生物生物量碳氮含量均表现为逐渐下降的趋势;类似地,土壤净氮矿化速率、硝化速率也逐渐降低。在20 ℃和60% SSM时,土壤净氮矿化速率表现为典型草地 (0.715 mg N kg^-1 d^-1) > 过渡草地 (0.507 mg N kg^-1 d^-1) > 荒漠草地 (0.134 mg N kg^-1 d^-1);相反,温度敏感性却逐渐升高,温度敏感性与基质质量指数呈负相关。草地类型和水分对于土壤净氮矿化速率、硝化速率具有显著影响,且二者间具有显著的交互效应。包含温度和水分的双因素模型可很好地拟合土壤氮矿化速率的变化趋势(P < 0.0001),二者可共同解释土壤硝化速率92%-96%的变异。土壤氮矿化沿着草地演替呈现出很好的空间格局、并与温度和水分具有密切关系,为解释内蒙古草地空间分布格局提供了理论基础。     

土壤碳矿化潜力对沙坡头人工固沙植被演变的响应

土壤呼吸是土壤有机C矿化分解,释放无机养分的重要生物化学过程。本研究通过实验室培养的方法,分析了沙坡头地区人工固沙区不同固沙年限土壤碳矿化潜力的变化。经过103d的室内培养,土壤CO2-C的释放量表现为55龄>47龄>30龄>24龄>21龄>流动沙丘,在垂直方向上表现为0～5cm>5～10cm>10～20cm。而流沙区土壤碳矿化潜力为10～20cm土层最高。不同固沙年限土壤碳矿化潜力、全氮、有机碳、电导率有明显的差异,均表现为随植被恢复年限的延长而增加,随深度的增加而递减。相关性分析表明,土壤碳矿化潜力与土壤有机碳、总氮、C/N、pH、电导率、温度、土壤水分含量呈极显著相关,土壤各环境因子之间亦呈极显著相关。土壤养分含量随着恢复时间的延长而得到明显的改善,土壤碳矿化潜力与土壤养分状况改善程度一致。人工固沙植被的建立促进了土壤微生物活性,通过潜在的土壤呼吸得到表征。植被恢复和凋落物积累使土壤免遭风蚀,显著增加了土壤有机质的输入,因而显著作用于大气C的固存。     

Der Einfluß von Vitamin A und β-Carotin auf die Fruchtbarkeit bei suboptimalen und optimalen Vitamin-B-Gaben

Die vorliegenden Untersuchungen sind Bestandteil eines komplexen Forschungsprogramms zur Weiterentwicklung der energetischen Futterbewertung im Nettoenergie‐Fett‐System. Sie wurden mit dem Ziel durchgeführt, neue Ergebnisse zur Erfassung des Zusammenhanges zwischen Ort und Art der Nährstoffverdauung und der energetischen Verwertung von Rationen bei der Tierart Rind zu erarbeiten. Für 9 Rationsvarianten mit jeweils 3 Varianten der Stärkeherkunft (Gerste, Mais, Kartoffeln) und ihres Rationsanteiles (50, 25 und 10 %) wurde an weitgehend ausgewachsenen Ochsen mit Hilfe von duodenalen Brückenfisteln auf dem Ernährungsniveau 1.7 die ruminale Nährstoffverdaulichkeit gemessen. Bei Stärkeeinnahmen zwischen 484 und 2573 g je Tier und Tag wurden Mengen an ruminai und postruminal verdauter Stärke zwischen 444 und 2336 bzw. 10 und 284 g je Tier und Tag bestimmt. Für die organische Substanz, Stärke, wasserlöslichen Kohlenhydrate und N‐freien Reststoffe wurden hohe relative Anteile der ruminai verdauten an den scheinbar verdauten Nährstoffen mit Werten zwischen 78 und 88, 83 und 98, 93 und 97 bzw. 88 und 100% ermittelt.     

高敏C-反应蛋白在紫癜性肾炎患儿中的应用

目的：探讨血清高敏C-反应蛋白（hs-CRP）在儿童紫癜性肾炎（HSPN）临床分型与病理分级中的应用价值,为基层医院提供一个可评价HSPN患儿病情严重程度的实验室相关指标。方法：应用免疫比浊法检测210例HSPN患儿不同临床分型与病理分级中的血清hs-CRP的水平,并与住院的70例的正常儿童作对照组进行比较。采用Pearson秩相关分析得出HSPN患儿血清hs-CRP水平临床分型与及病理分级的关系。结果：HSPN患儿血清hs-CRP水平明显高于对照组（HSPN组6.4±3.5 mg/L,对照组0.7±0.1mg/L）,差异有统计学意义（t=1.021,P=0.003）。HSPN患儿的血清hs-CRP水平与其临床分型的严重程度存在正相关（r=0.913,P〈0.05）。而HSPN患儿血清hs-CRP水平与其病理分级的关系也呈正相关（r=0.901,P〈0.05）。结论：随着HSPN患儿临床分型与病理分级的增高,其血清hs-CRP水平显著升高,HSPN患儿血清hs-CRP水平与其临床分型和病理分级之间均呈显著正相关,检测HSPN患儿血清hs-CRP水平可预测其临床分型和病理分级的程度,即HSPN患儿血清hs-CRP水平越高提示其临床分型和病理分级越重,因此检测HSPN患儿血清hs-CRP水平有助于评估HSPN患儿的病情、治疗效果和预后情况。     

淋巴结转移阴性的胃癌患者临床病理特征及生存探讨

目的:探讨淋巴结转移阴性胃癌患者的临床病理特征以及预后影响因素。方法:收集2000年1月至2009年1月我院收治的胃癌患者325例,其中经病理检查显示淋巴结转移阴性的105例患者作为阴性组(LN-组),另229例阳性患者作为阳性组(LN+组),比较两组的临床病理特征及临床预后。结果:LN-组的肿瘤直径、浸润深度及术后化疗与LN+组比较差异显著(P0.05);LN-组的5年生存率为76.2%,显著高于LN+组的43.2%(P0.05)。未透浆膜的LN-患者3年、5年生存率显著高于浸透浆膜者,术后化疗的LN-患者5年生存率显著高于未化疗者(P0.05),肿瘤直径5 cm的LN-患者3、5年生存率显著高于≥5 cm者(P0.05)。单因素分析显示浸润深度、肿瘤大小及术后化疗与LN-胃癌患者的预后具有密切关系(P0.05)。COX多因素分析显示浸润深度是影响LN-胃癌患者临床预后的独立因素(P0.05)。结论:淋巴结转移阴性胃癌患者的病灶多位于中下部,男性多于女性,发病年龄多在60岁以内,肿瘤直径多不超过5 cm,浸润深度多未浸透浆膜,临床预后优于淋巴结转移阳性胃癌患者,浸润深度是影响淋巴结转移阴性胃癌患者临床预后的独立因素。     

Intensified plant N and C pool with more available nitrogen under experimental warming in an alpine meadow ecosystem

Nitrogen (N) availability is projected to increase in a warming climate. But whether the more available N is immobilized by microbes (thus stimulates soil carbon (C) decomposition), or is absorbed by plants (thus intensifies C uptake) remains unknown in the alpine meadow ecosystem. Infrared heaters were used to simulate climate warming with a paired experimental design. Soil ammonification, nitrification, and net mineralization were obtained by in situ incubation in a permafrost region of the Qinghai‐Tibet Plateau (QTP). Available N significantly increased due to the stimulation of net nitrification and mineralization in 0–30 cm soil layer. Microbes immobilized N in the end of growing season in both warming and control plots. The magnitude of immobilized N was lower in the warming plots. The root N concentration significantly reduced, but root N pool intensified due to the significant increase in root biomass in the warming treatment. Our results suggest that a warming‐induced increase in biomass is the major N sink and will continue to stimulate plant growth until plant N saturation, which could sustain the positive warming effect on ecosystem productivity.