NH4NO3浓度对吊兰叶片形态和生理生化指标的影响

采用沙培的方式,使用NH4NO3对幼苗进行5种不同的氮素浓度（1 mmol/L,5 mmol/L,10 mmol/L,15 mmol/L,20 mmol/L）处理,每1种处理方式重复5次.通过对吊兰叶片叶长、叶宽、叶绿素含量,可溶性蛋白含量,SOD活性等一系列指标的测定,优选出最适合吊兰生长的氮素浓度.结果表明：5 mmol/L处理的吊兰叶片,其叶片长度、宽度、叶绿素以及蛋白质含量和SOD,POD活性都明显高于其它浓度.     

The equine alveolar macrophage: Functional and phenotypic comparisons with peritoneal macrophages

Alveolar macrophages (AMs) constitute the first line of defence in the lung of all species, playing a crucial role in the regulation of immune responses to inhaled pathogens. A detailed understanding of the function and phenotype of AMs is a necessary pre-requisite to both elucidating their role in preventing opportunistic bacterial colonisation of the lower respiratory tract and developing appropriate preventative strategies. The purpose of the study was to characterise this important innate immune cell at the tissue level by making functional and phenotypic comparisons with peritoneal macrophages (PMs). We hypothesised that the tissue of origin determines a unique phenotype of AMs, which may constitute an appropriate therapeutic target for certain equine respiratory diseases. Macrophages isolated from the lung and the peritoneal cavity of 9 horses were stimulated with various toll like receptor (TLR) ligands and the production of nitrite, tumour necrosis factor alpha (TNFα), interleukin (IL) 10 and indoleamine 2,3-dioxygenase (IDO) were measured by the Griess reaction and enzyme linked immunosorbent assay (ELISA) and/or quantitative polymerase chain reaction, respectively. Cells were also compared on the basis of phagocytic-capacity and the expression of several cell surface markers. AMs, but not PMs, demonstrated increased TNFα release following stimulation with LPS, polyinosinic polycytidylic acid (Poly IC) and heat-killed Salmonella typhinurium and increased TNFα and IDO mRNA expression when stimulated with LPS. AMs showed high expression of the specific macrophage markers cluster of differentiation (CD) 14, CD163 and TLR4, whereas PMs showed high expression of TLR4 only. AMs, but not PMs, demonstrated efficient phagocytic activity. Our results demonstrate that AMs are more active than PMs when stimulated with various pro-inflammatory ligands, thus supporting the importance of the local microenvironment in the activation status of the macrophage. This information provides a valuable knowledge base on which to improve our understanding of the role of macrophages and their microenvironment in equine innate immunity.     

阿氏节杆菌菌株DA-1的厌氧反硝化研究

阿氏节杆菌(Arthrobacter arilaitensis)菌株DA-1是一株氨氮降解细菌,且在有氧及厌氧条件下均具有反硝化作用.通过对该菌的厌氧反硝化条件进行优化,考察了碳源、C/N、温度和初始pH对其反硝化作用的影响.结果表明,乙酸钠为最佳的反硝化碳源,当C/N为10∶1、温度为30℃、初始pH为7.0时,A.arilaitensis DA-1能获得最高的反硝化效率,在该条件下,向硝酸盐模拟污水(NO3--N浓度为25mg/L)中接入2％(V/V)的A.arilaitensis DA-1菌剂,经过48 h的静置培养,N03--N的去除量达到了20.13 mg/L,而此时NO2--N积累浓度仅为0.56 mg/L.     

一氧化氮信号途径参与草地早熟禾耐镉机制的研究

为探究信号分子一氧化氮（Nitric Oxide,NO）对镉（Cadmium,Cd）胁迫下草地早熟禾（Poa pratensis）的调控机理,揭示草地早熟禾耐镉机理,本试验采取根施1000 μmol·L^-1氯化镉（Cadmium chloride hemidihydrate,CdCl₂·2.5H₂O）及叶面喷施NO合成酶抑制剂（L-硝基精氨酸甲酯（L-NAME））、硝酸还原酶抑制剂（钨酸钠（Tungstate））、NO清除剂（4-羧基苯-4,4,5,5-四甲基咪唑-1-氧-3-氧化物（cPTIO））处理草地早熟禾叶片,研究不同处理对草地早熟禾幼苗叶片生长及抗氧化酶活性、光合色素含量、丙二醛（Malondialdehyde,MDA）、游离脯氨酸、内源NO含量的影响。研究发现,镉胁迫下草地早熟禾内源NO含量增加,添加一氧化氮清除剂、一氧化氮合酶抑制剂、硝酸还原酶抑制剂可增加超氧化物歧化酶（Superoxide Dismutase,SOD）、过氧化氢酶（Catalase,CAT）、抗坏血酸过氧化物酶（Ascorbate peroxidase,APX）活性,降低过氧化物酶（Peroxidase,POD）活性,增加叶绿素a、叶绿素b、类胡萝卜素、丙二醛含量,降低脯氨酸（Proline,Pro）含量、叶片相对含水量（Relative Water Content,RWC）和干物质量（Dry matter content）。结果表明,一氧化氮合酶途径可能是早熟禾合成NO的主要途径,内源NO可通过调节抗氧化酶活性、光合色素含量、游离脯氨酸含量等途径缓解镉胁迫。     

Extraction of mobile element fractions in forest soils using ammonium nitrate and ammonium chloride

The extraction of earth alkaline and alkali metals (Ca, Mg, K, Na), heavy metals (Mn, Fe, Cu, Zn, Cd, Pb) and Al by 1 M NH₄NO₃ and 0.5 M NH₄Cl was compared for soil samples (texture: silt loam, clay loam) with a wide range of pH(CaCl₂) and organic carbon (OC) from a forest area in W Germany. For each of these elements, close and highly significant correlations could be observed between the results from both methods in organic and mineral soil horizons. The contents of the base cations were almost convertible one‐to‐one. However, for all heavy metals NH₄Cl extracted clearly larger amounts, which was mainly due to their tendency to form soluble chloro complexes with chloride ions from the NH₄Cl solution. This tendency is very distinct in the case of Cd, Pb, and Fe, but also influences the results of Mn and Zn. In the case of Cd and Mn, and to a lower degree also in the case of Pb, Fe, and Zn, the effect of the chloro complexes shows a significant pH dependency. Especially for Cd, but also for Pb, Fe, Mn, Zn, the agreement between both methods increased, when pH(CaCl₂) values and/or contents of OC were taken into account. In comparison to NH₄Cl, NH₄NO₃ proved to be chemically less reactive and, thus, more suitable for the extraction of comparable fractions of mobile heavy metals. Since both methods lead to similar and closely correlated results with regard to base cations and Al, the use of NH₄NO₃ is also recommended for the extraction of mobile/exchangeable alkali, earth alkaline, and Al ions in soils and for the estimation of their contribution to the effective cation‐exchange capacity (CEC). Consequently, we suggest to determine the mobile/exchangeable fraction of all elements using the NH₄NO₃ method. However, the applicability of the NH₄NO₃ method to other soils still needs to be investigated.     

急性冰水应激对犬血清甲状腺激素、NO及Na、K、Cl离子含量的影响

通过测定急性冰水应激时犬血清甲状腺激素、NO及Na、K、Cl离子含量的变化,来评价急性冰水应激对犬体内能量代谢、氧化应激状态及电解质平衡的影响。试验选取8只成年杂种犬,保定躯干浸于冰水混合物中(0~4)℃,使其进行冰水浴30 min。分别于处理前0.5 h(-0.5 h)及处理后0、0.5、1、3、7 h进行桡静脉采血2 mL,分离血清后测定其甲状腺激素(T3、T4)、一氧化氮(NO)及Na^+、K^+、Cl^-的含量。结果表明:与-0.5 h(处理前)相比,处理后各时间点犬血清T3含量无显著变化(P>0.05),与-0.5 h及0、3、7 h相比,血清T4含量在0.5 h时显著降低(P<0.05);血清NO含量在处理后各时间点均显著高于处理前0.5 h(P<0.05),且1、3、7 h显著高于0、0.5 h(P<0.05);与-0.5 h相比,处理后3 h血清Na^+含量显著升高(P<0.05),血清K^+含量处理后0 h显著高于1、3、7 h(P<0.05),血清Cl^-含量处理前后差异不显著(P>0.05)。研究表明,急性短期冰水应激会扰乱机体的能量代谢状态、降低T4含量,同时造成氧化应激损伤、使NO含量大量升高;引起血清Na、K离子平衡紊乱、含量升高,提示临床上人或动物发生冰水应激时应及时纠正机体能量代谢、氧化还原平衡和电解质的紊乱。     

沙冬青种子总黄酮活性成分分析及对小鼠免疫功能的影响

本研究旨在通过高效液相色谱-质谱联用（LC-MS）和高效液相色谱（HPLC）技术对沙冬青种子总黄酮主要活性成分进行分析及含量测定,并探索沙冬青种子总黄酮对小鼠免疫功能的影响。采用LC-MS分析沙冬青种子总黄酮主要活性成分,在HPLC优化条件下用标准品对主要活性成分的含量进行测定;采用MTT法和ELISA试剂盒检测沙冬青种子总黄酮对小鼠脾淋巴细胞增殖和IL-2、IL-4生成的影响,以及总黄酮对肝脏KCs细胞的增殖和NO、NOS含量的影响;运用定量溶血分光光度法和血清溶血素HC₅₀测定法检测不同浓度沙冬青种子总黄酮对小鼠抗体生成细胞和血清溶血素的影响。结果显示,沙冬青种子总黄酮中主要活性成分为芒柄花素、7,3'-二羟基-4'-甲氧基黄酮、黄豆黄素和穗花杉双黄酮,其中,芒柄花素含量高达52.48%,其次为7,3'-二羟基-4'-甲氧基黄酮,含量为11.20%。在2.5~100 μg/mL总黄酮作用下,沙冬青种子总黄酮对脾淋巴细胞的增殖和细胞因子的产生具有明显的促进作用,尤其在20 μg/mL时脾淋巴细胞增殖率和IL-2的分泌量分别高达217.62%和159.661 pg/mL,与空白对照组相比差异极显著（P<0.01）;在40 μg/mL时,脾淋巴细胞IL-4的分泌量高达149.274 pg/mL,极显著高于空白对照组（P<0.01）。沙冬青种子总黄酮对KCs细胞的增殖及NO和NOS分泌也具有明显促进作用,在60 μg/mL时,与空白对照组相比,KCs细胞增殖率达67.77%,NO和NOS的分泌量分别达180.106和29.942 μmol/L（P<0.01）。此外,沙冬青种子总黄酮能明显促进小鼠体内抗体生成细胞和血清溶血素含量的增加。综上表明,沙冬青种子总黄酮主要活性成分为芒柄花素和7,3'-二羟基-4'-甲氧基黄酮。沙冬青种子总黄酮对小鼠免疫活性细胞的增殖、相关免疫细胞因子和抗体的产生与释放等都具有显著的促进作用。     

不同水稻品种对NO-3 同化差异的比较

采用水培方法测定了不同形态氮素下 4个品种水稻 (汕优 63、扬稻 6号、泗优 917、农垦 57) 的生长量及其水稻苗期硝酸还原酶活性 (NRA) 和谷氨酰胺合成酶活性 (GSA)。结果表明, 1mmol·L-1 NH 4 培养下, 水稻生长无明显差异, 而 1mmol·L-1 NO-3 培养 28d后, 各品种水稻生长差异显著, 其中, 扬稻 6号生长最优, 农垦 57最差; 籼稻体内的NRA和GSA比粳稻更高, 其中籼稻叶片的NRA比粳稻高出 58 7%, GSA高出 34 6%, 籼稻根系GSA是粳稻根系的 3 2倍, 说明籼稻对NO-3 的吸收利用优于粳稻。     

不同形态氮素及铵硝比例对咖啡氮吸收和生长的影响

为提高咖啡氮肥肥料有效性,采用溶液培养的方法,研究NH4+和NO3- 2种不同形态氮吸收速率、5种铵硝比例(10∶0、7∶3、5∶5、3∶7、0∶10)对咖啡生长及其氮素利用的影响。结果表明,不同形态氮素对咖啡的生长影响差异显著,铵硝混合营养下咖啡的生长明显优于单一形态氮素处理。在单一形态氮素条件下,咖啡对NH4+的最大吸收速率大于对NO3-的最大吸收速率;当2种形态氮素同时存在时,铵态氮会抑制硝态氮的吸收,硝态氮促进铵态氮的吸收;铵态氮促进地上部分生长,但浓度过高反而抑制地上部分生长;硝态氮的增加有利于根系的生长,但抑制了咖啡地上部分的生长。因此,在咖啡苗期,铵硝比例控制在7∶3～3∶7有利于咖啡生长。     

杏鲍菇多糖的提取及其对RAW264.7细胞吞噬活性·NO释放的影响

[目的]以杏鲍菇多糖的理化性质作为综合评价指标,提取杏鲍菇多糖,并研究其对RAW264.7细胞吞噬活性的作用及对NO释放的影响。[方法]采用水煎煮提取、超声提取、90℃热水浸提、70℃热水浸提、50℃热水浸提提取杏鲍菇多糖,并采用苯酚-硫酸法、间羟基联苯法以及Lowry法测定其总糖、酸性糖和蛋白质含量;采用PMP柱前衍生化法和HPGPC法测定其组成糖和分子量分布;采用吞噬中性红法和Griess试剂盒法测定其吞噬能力及NO的释放量。[结果]试验得出,50℃热水浸提提取的多糖总糖含量最高,超声提取的多糖蛋白质含量最高,各组提取酸性糖含量相当且均较低,水煎煮提取多糖中的单糖种类最少、分子量最低。综合比较,90℃热水浸提为杏鲍菇多糖的最佳提取方法;在质量浓度为25~100μg/m L,能够极显著地促进细胞吞噬中性红的能力和NO的释放。[结论]90℃热水浸提水煎煮为杏鲍菇多糖的最佳提取方法,一定浓度的杏鲍菇多糖能够提高RAW264.7细胞的吞噬活性,并能促进NO的释放,提示杏鲍菇多糖具有一定的免疫活性。