单增李斯特菌感染RAW264.7细胞转录组差异表达分析

为了探讨单核细胞增生李斯特菌（Listeria monocytogenes,LM）感染前后RAW264.7细胞基因表达谱变化,筛选关键功能基因和通路,为探究其致病机制奠定基础,本研究以感染复数（MOI）10的LM 感染RAW264.7细胞6 h后,提取细胞RNA进行转录组测序,选取感染组与对照组相比差异倍数≥1.5且P<0.05的差异表达基因进行GO和KEGG富集分析,并通过RT-qPCR验证部分差异基因。转录组结果显示RAW264.7细胞在感染LM前后有1 782 个基因差异表达,其中上调为989,下调为793。GO分析结果主要涉及免疫系统过程、免疫反应、细胞程序性死亡调控等。KEGG分析结果显示,显著富集的通路有细胞因子-受体相互作用、肿瘤坏死因子信号通路、IL-17信号通路等。RT-qPCR验证试验结果显示,随机选择的11 个显著差异基因表达倍数趋势与测序结果一致。该研究为进一步深入研究LM的致病机制奠定了基础。     

橄榄苦苷改善db/db小鼠糖尿病的肝脏转录组学及生物信息学分析

通过转录组学及其生物信息学分析,研究了橄榄苦苷缓解db/db小鼠糖尿病的肝脏差异表达基因及相关信号通路。结果发现与db/db对照组相比,橄榄苦苷处理组的539 个基因发生显著变化,其中450 个基因显著上调,89 个基因显著下调。将上调和下调的差异表达基因在基因本体论数据库中注释,这些差异表达基因主要在细胞过程、细胞部分和结合中分布。京都基因与基因组百科全书通路富集分析结果显示上调的差异表达基因主要富集在磷酸肌醇3-激酶-蛋白激酶B信号通路途径,该通路共涉及27 个差异表达基因;下调的差异表达基因主要富集在花生四烯酸代谢和真核生物中核糖体的生物发生信号通路途径,分别涉及4 个差异表达基因。本研究为进一步阐明橄榄苦苷缓解2型糖尿病的分子机制奠定了基础。     

基于转录组学分析日本沼虾原肌球蛋白对小鼠巨噬细胞极化的影响

为研究日本沼虾原肌球蛋白对巨噬细胞极化的影响,运用转录组学分析原肌球蛋白诱导的巨噬细胞极化过程中基因表达的差异性。结果表明:与磷酸盐缓冲溶液组(PBS组)相比,原肌球蛋白诱导组(TM组)有69个基因表达上调,154个基因表达下调,富集到180条通路;TM组相较于脂多糖和IFN-γ联合诱导M1型巨噬细胞组(LPSIFN组)有1346个基因表达上调,1360个基因下调,富集到308条通路;TM组与IL-4诱导M2型巨噬细胞组(IL4组)相比,有455个基因上调,446个基因下调,富集到269条通路。根据KEGG结果选择5条可能涉及巨噬细胞极化的信号通路,包括NOD样受体信号通路、Jak-STAT信号通路、NF-κB信号通路、Toll样受体信号通路和PI3K/Akt信号通路,对这5条通路中的NOD2、TLR2、AKT3、NLRP3、Caspase-1、CD14、Lat、Myd88、NFKBIA和STAT1基因的表达进行验证,检测到TM组NOD2、TLR2、NLRP3、STAT1这4个基因表达量相较于PBS组显著升高,与这4个基因相关的NOD样受体信号通路、Jak-STAT信号通路、Toll样受体信号通路参与原肌球蛋白诱导巨噬细胞极化的过程。研究旨在为进一步分析巨噬细胞极化与食物过敏的关系提供理论依据。     

反油酸诱导人脐静脉内皮细胞转录组学及生物信息学分析

以反油酸诱导人脐静脉内皮细胞后,结合转录组学分析及其生物信息学分析,在转录水平上对相关差异表达基因进行研究。结果表明：共检测到95?条基因显著变化,其中31?条基因上调,64?条基因下调。并通过生物信息学分析,其中参与生物进程显著性差异表达基因共772?条,关于细胞组成的基因显著表达性差异基因共46?条,关于分子功能的基因显著表达性差异基因共122?条。共检测到93?条信号通路,其中显著变化的有11?条,其中包括萜类骨架生物合成通路、类固醇生物合成通路和脂肪酸代谢通路等。根据蛋白质基因互作通路图,重点分析了同时参与不同通路的靶基因,包括脂肪酸合成酶、STAT4转录因子及IL-23细胞因子等。     

芥子酸对油酸诱导HepG2细胞脂毒性模型 差异表达基因的影响

菜籽油富含酚酸,其中芥子酸具有较强的抗氧化能力,能够清除氧自由基,提高细胞氧化应激反应能力。以油酸诱导HepG2细胞构建脂毒性模型,通过RNA-seq分析芥子酸干预对HepG2细胞差异表达基因的影响,并对HepG2细胞中差异表达基因进行了GO分类和KEGG富集分析。结果表明：对照组、模型组、实验组、模型实验组间均存在差异表达基因,与对照组相比,模型组、模型实验组和实验组中下调基因多于上调基因。GO功能分类分析发现油酸和芥子酸主要从细胞过程、刺激反应、催化活性方面影响细胞物质代谢和酶的催化基因表达。在KEGG通路富集中,油酸和芥子酸干预改变了HepG2细胞相关有丝分裂原活化蛋白激酶信号通路、松弛素信号通路和白细胞介素17信号通路等多个通路,说明芥子酸对细胞有机系统有一定的影响,并且可以通过调节PPAR信号通路来刺激PPARγ的活性从而影响脂肪的代谢与沉积。     

烟草叶片在二氯喹啉酸胁迫下的蛋白组学分析

为明确二氯喹啉酸对烟草药害的机理,采用同位素标记相对和绝对定量(i TRAQ)技术分析了二氯喹啉酸处理7 d后烟草畸形叶片较正常叶片的蛋白组学变化。本研究共鉴定出65个差异蛋白,其中表达量上调的23个,下调的42个,它们具有结合、催化和转运等分子功能。分别参与光合作用、防御/抗胁迫、代谢等细胞过程,荧光定量PCR验证结果与蛋白组学数据一致。ABA合成通路中的ABA2及NCED基因能够响应二氯喹啉酸胁迫信号,分别在处理后48和24 h达到表达高峰,处理后72 h下调至较低水平。根据这种上调表达后又迅速下调的基因表达模式,推测烟草体内存在一种应激机制抵抗二氯喹啉酸胁迫。本研究首次在烟草中采用蛋白组学的方法分析二氯喹啉酸胁迫下基因的表达变化,并获得重要候选基因。     

低钠盐饮食对胰岛素信号通路相关基因网络状态的影响

研究低钠盐饮食对高血压与肥胖并发者血液中胰岛素信号通路相关基因表达的影响,初步找到表达发生变化的基因间相互关联的网络状态.选择3名高血压与肥胖并发者,分别对他们食用低钠盐前后的血样做胰岛素信号通路表达谱芯片,共6张.以每个受试者食用前所做芯片图做对照,找出其食用后表达发生变化的基因.在检测的144个基因中,得到2次以上重复一致的异常表达基因共26条:上调基因6个,下调基因20个.食用低钠盐对胰岛素信号传导通路中的Focal Adhesion通路和PI3K途径有直接的影响,上述结果从血液基因表达的层面和局部基因网络的角度提供了食低钠盐缓减胰岛素抵抗的初步证据.     

离子束重组异常汉逊酵母菌Ar_Han0458的转录组学分析及谷胱甘肽代谢途径富集

研究可稳定遗传的重组异常汉逊酵母Ar_Han0458在发酵过程中的差异基因表达信息及与谷胱甘肽代谢相关基因的表达变化。利用生物信息学方法分析重组异常汉逊酵母Ar_Han0458的5个发酵时间点的RNA-seq数据。重组异常汉逊酵母Ar_Han0458各发酵时间与0 h相比,下调表达基因数量多于上调表达;除发酵96和72 h外,相邻时间点的差异表达基因数目均以下调为主;发酵48和72 h时的基因表达谱相近。各发酵时间点间差异表达基因显著富集的GO功能均以细胞过程、细胞和催化活性为主。KEGG富集分析显示,10个差异表达基因参与了谷胱甘肽的代谢,谷胱甘肽的合成速率分别在24和96 h达到峰值,与实验测得胞外GSH产量变化趋势一致。重组异常汉逊酵母Ar_Han0458在发酵96 h时差异表达基因数目以上调为主,且谷胱甘肽合成速率最快。该研究为重组异常汉逊酵母的代谢调控研究及其分子育种提供了理论依据。     

转录组学分析空气对泡菜“生花”酵母菌的影响机制

泡菜坛/池的密封性是影响泡菜“生花”的关键因素,为了明确空气对“生花”酵母菌的影响,运用转录组学和生物信息学分析探索影响酵母菌“生花”的关键基因。研究结果表明,有和无空气条件下培养的“生花”酵母菌共筛选到183个差异基因,其中42个基因显著上调,141个基因显著下调(P<0.05)。差异表达基因主要富集在葡萄糖苷酶活性、阿拉伯糖脱氢酶活性和核糖体亚基等功能。差异基因富集到34个通路,其中核糖体通路是差异最显著的通路,包含RPL4、RPL10、RPSA等19个基因上调和RPS2、RPS15基因下调。脂肪酸合成通路和不饱和脂肪酸合成通路中差异基因FAD2上调,FAS1、FAS2、SCD等6个基因下调。经实时荧光定量PCR实验验证,在有空气培养条件下,关键基因FAD2、RPL4和RPL10较无空气培养显著上调(P<0.05),与转录组预测结果一致。该研究结果能够为泡菜“生花”酵母菌的“生花”机理以及泡菜品质控制提供理论基础。     

高通量基因表达谱的应用

基因表达谱可以描绘特定情况下组织、细胞中所表达的全套基因及其丰度,它从mRNA水平上反映出组织或细胞特异性表型和表达模式.目前基于高通量的基因表达谱主要有基因芯片和数字基因表达谱(DGE).本文主要介绍了基因芯片和DGE的原理、特点,并对其在作物科学、动物科学和乳品科学中的应用做了简要列举.     

Arsenic(III) oxidation by iron(VI) (ferrate) and subsequent removal of arsenic(V) by iron(III) coagulation

We investigated the stoichiometry, kinetics, and mechanism of arsenite [As(III)] oxidation by ferrate [Fe(VI)] and performed arsenic removal tests using Fe(VI) as both an oxidant and a coagulant. As(III) was oxidized to As(V) (arsenate) by Fe(VI), with a stoichiometry of 3:2 [As(III):Fe(VI)]. Kinetic studies showed that the reaction of As(III) with Fe(VI) was first-order with respect to both reactants, and its observed second-order rate constant at 25 degrees C decreased nonlinearly from (3.54 +/- 0.24) x 10(5) to (1.23 +/- 0.01) x 10(3) M(-1) s(-1) with an increase of pH from 8.4 to 12.9. A reaction mechanism by oxygen transfer has been proposed for the oxidation of As(III) by Fe(VI). Arsenic removal tests with river water showed that, with minimum 2.0 mg L(-1) Fe(VI), the arsenic concentration can be lowered from an initial 517 to below 50 microg L(-1), which is the regulation level for As in Bangladesh. From this result, Fe(VI) was demonstrated to be very effective in the removal of arsenic species from water at a relatively low dose level (2.0 mg L(-1)). In addition, the combined use of a small amount of Fe(VI) (below 0.5 mg L(-1)) and Fe(III) as a major coagulant was found to be a practical and effective method for arsenic removal.     

Mind the (yield) gap(s)

This paper explores the origin of the notion of ??yield gap?? and its use as a framing device for agricultural policy in sub-Saharan Africa. The argument is that while the yield gap of policy discourse provides a simple and powerful framing device, it is most often used without the discipline or caveats associated with the best examples of its use in crop production ecology and microeconomics. This argument is developed by examining how yield gap is used in a selection of recent and influential agricultural policy documents. The message for policy makers and others is clear: ??mind the (yield) gap(s)??, for they are seldom what they appear.     

Iron(VI) and iron(V) oxidation of copper(I) cyanide

Copper(Il) cyanide (Cu(CN)4(3-)) in the gold mine industry presentsthe biggest concern in cyanide management because it is much more stable than free cyanide. Cu(CN)4(3-) is highlytoxic to aquatic life; therefore, environmentally friendly techniques are required for the removal of Cu(CN)4(3-) from gold mine effluent. The oxidation of Cu(CN)4(3-) by iron-(VI) (FeVIO4(2-), Fe(VI)) and iron(V) (FeVO4(3-), Fe(V)) was studied using stopped-flow and premix pulse radiolysis techniques. The stoichiometry with Fe(VI) was determined to be 5HFeO(4-) + Cu(CN)4(3-) + 8H2O - > 5Fe(OH)3 + Cu2+ + 4CNO- +3/202 + 6OH-. The rate law for the oxidation of Cu(CN)4(3-) by Fe(VI) was found to be first-order with each reactant. The rates decreased with increasing pH and were mostly related to a decrease in concentration of reactive protonated Fe(VI) species, HFeO4-. A mechanism is proposed that agrees with the observed reaction stoichiometry and rate law. The rate constant for the oxidation of Cu(CN)4(3-) by Fe(V) was determined at pH 12.0 as 1.35 +/- 0.02 x 10(7) M(-1) s(-1), which is approximately 3 orders of magnitude larger than Fe(VI). Results indicate that Fe(VI) is highly efficient for removal of cyanides in gold mill effluent.     

Supercritical fluid extraction (SFE) of 4(5)-methylimidazole (4-MeI) and 2-acetyl-4(5)-(1,2,3,4)-tetrahydroxybutyl-imidazole (THI) from ground-coffee with high-performance liquid chromatographic-electrospray mass spectrometric quantification (HPLC/ESI-MS).

Two polar analytes, 4(5)-methylimidazole (4-MeI) and 2-acetyl-4(5)-(1,2,3,4)-tetrahydroxybutyl-imidazole (THI), were extracted with supercritical carbon dioxide (CO2) modified with aqueous methanol. The method was applied to a roasted coffee powder with good recovery rates. Method efficiency was compared with that of solid-phase extraction using SCX Disc cartridges and validated for spiked solid matrix. The analytes were determined using isocratic liquid chromatography-mass spectrometry (LC/MS) on an Atlantis HILIC Silica column (150 x 2.1 mm, 3 microm) with 80% methanol and 20% 0.01 mol l-1 ammonium formate as the mobile phase. The limit of quantification was around 1.5 pg for 4-MeI and 2.0 pg for THI. The linearity of the calibration curves was satisfactory as indicated by correlation coefficients of >0.999. The coefficient of variation for the intra-day and inter-day precisions was <4% (n = 6). Accuracy was in the range 98-101%; recovery rates were > or = 98 and > or = 99% for THI and 4-MeI, respectively. Several samples of Arabica coffee from various locations and commercially available 'off-the-shelf' coffee products (Arabica/Robusta mixtures) were analysed to test the method.     

Reduction of Tc(VII) by Fe(II) sorbed on Al (hydr)oxides

Under oxic conditions, Tc exists as the soluble, weakly sorbing pertechnetate [TcO4-] anion. The reduced form of technetium, Tc(IV), is stable in anoxic environments and is sparingly soluble as TcO2 x nH2O(s). Here we investigate the heterogeneous reduction of Tc(VII) by Fe(II) adsorbed on Al (hydr)oxides [diaspore (alpha-AlOOH) and corundum (alpha-Al2O3)]. Experiments were performed to study the kinetics of Tc(VII) reduction, examine changes in Fe surface speciation during Tc(VII) reduction (M?ssbauer spectroscopy), and identify the nature of Tc(IV)-containing reaction products (X-ray absorption spectroscopy). We found that Tc(VII) was completely reduced by adsorbed Fe(II) within 11 (diaspore suspension) and 4 days (corundum suspension). M?ssbauer measurements revealed thatthe Fe(II) signal became less intense with Tc(VII) reduction and was accompanied by an increase in the intensity of the Fe(III) doublet and magnetically ordered Fe(III) sextet signals. Tc-EXAFS spectroscopy revealed that the final heterogeneous redox product on corundum was similar to Tc(IV) oxyhydroxide, TcO2 x nH2O.     

表没食子儿茶素没食子酸酯的甲基化分子修饰

研究表没食子儿茶素没食子酸酯(EGCG)的甲基化分子修饰。以碘甲烷作为甲基供体,采用化学合成方法研究EGCG 的甲基化分子修饰,并通过HPLC-MS 和NMR 等对反应产物进行结构鉴定。结果表明：采用化学合成方法能有效完成EGCG 的甲基化分子修饰,并分离鉴定出5 个EGCG 甲基化衍生物,分别为4″-Me-EGCG、4′,4″-di-Me-EGCG、5,3′,4′,5′,3″,4″,5″-hepta-Me-EGCG、5,7,3′,4′,3″,4″,5″-hepta-Me-EGCG、5,7,3′,4′,5′,3″,4″,5″-octa-Me-EGCG。     

Attractancy of diethyl ether extracts of Arachis hypogaea (Linnaeus) (Papilionaceae), Citrullus lanatus (Thunberg) (Cucurbitaceae) and Irvingia gabonensis var. excelsa (Baillon) (Irvingiaceae) to Oryzaephilus mercator (Fauvel) (Coleoptera: Silvanidae)

Volatiles of the diethyl ether extracts of I. gabonensis, C. lanatus and A. hypogaea attracted 1–7 and 28–42-day-old adult O. mercator when tested in a two-choice pitfall Petri dish bioassay. For both age groups, a significant difference was observed in response between fed and starved adults to the different extracts. Differences in starvation time did not affect responses of individuals in the 1–7-day-old group; a significant effect was observed in the 28–42-day-old group.

Adults, 28–42-day-old were significantly more responsive to virtually all extracts than 1–7-day-old adults. Responses of adults in each age group to the different extracts were not significantly different. The time of trial within the period 0900–1900 h did not significantly affect the response of adults in both age groups, starved for 2 days, to any extract.     

优质农业生产管理指导手册(二)

4病虫害综合防治 病虫害综合防治(IPM)是烟叶生产的一项系统体系,利用病虫害预测预报系统,提供最好的病虫害综合防治决策,重点是协调应用一切可行的办法.病虫害综合防治并不意味着完全排除使用农药,而是在使用其它办法不能使病虫害的种群数保持在可接受的范围时适当使用农药.施用农药的地方应确保安全,并遵守一切法律法规.     

优质农业生产管理指导手册(三)

7 烘烤（调制）和烤房管理 烟叶的正确烘烤和烤房管理对优化烟叶的产量、质量和产值来说是非常关键的因素。烟叶烘烤是保持烟叶产量、质量的最后一道环节。烟叶质量会因烘烤不当而降低，而正确进行烘烤能确保烟叶质量的稳定。然而，若将劣质的鲜烟叶或烟株放入烤房，则不可能提高最终的烟叶质量。