猪苓菌丝细胞活性氧的产生及其种类

用液体发酵的蜜环菌菌丝、菌丝细胞壁及发酵液作为激发子,分别处理猪苓菌丝,均可诱导猪苓菌丝活性氧的产生。活性氧产生量与激发子浓度具相关性。超氧化物歧化酶（SOD）、过氧化氢酶(CAT)、甘露醇均可在一定程度上抑制活性氧的产生,证明活性氧种类包括过氧化氢（H     

蜜环菌激发子诱导猪苓细胞产生活性氧及其相关酶的变化*

研究了蜜环菌激发子处理猪苓细胞后活性氧产生及相关酶的变化。结果表明 :蜜环菌激发子分别处理猪苓菌丝和菌核后 ,都能引起活性氧迸发 ,且出现两个迸发高峰 ,高峰期分别在加入激发子后约 1 0min和 90min。与此同时 ,猪苓菌丝酶活性也发生相应变化 ,超氧化物岐化酶 (SOD)、过氧化物酶 (POD)活性下降 ,过氧化氢酶 (CAT)活性没有变化 ,而苯丙氨酸解氨酶 (PAL)活性出现先下降后上升的趋势。     

密环菌激发子诱导猪苓细胞产生活性氧及其相关酶的变化

研究了蜜环菌激发子处理猪苓细胞后活性氧及产生及相关酶的变化。结果表明：蜜环菌激发子分别处理猪铃菌丝和菌核后,都能引起活性氧迸发,且出现两个迸发高峰,高峰期分别在加入激发子后约10min和90min。与此同时,猪铃菌丝酶活性也发生相应变化,超氧化物歧化酶（SOD）、过氧化物酶（POD）活性下降,过氧化氢酶（CAT）活性没有变化,而苯丙氨酸解氨酶（PAL）活性出现先下降后上升的趋势。     

H2O2致WB-F344细胞内活性氧的产生及机理

以双氢罗丹明123（DHR123）作为荧光探针,采用激光共聚焦扫描显微镜研究小剂量（800nmol/L)H2O2诱导大鼠肝卵细胞株WB－F344细胞内活性氧产生的动态变化过程及其机理。结果发现：（1）小剂量H2O2的一次作用可以引起胞内活性氧的产生;（2）胞内活性氧清除剂N－乙酰－L－半胱氨酸（NAC）处理2h时后,再加入小剂量H2O2,发现胞内活性氧的产生明显减少;（3）用广谱的蛋白激酶抑制剂2－氨基嘌呤（2－AP）、Ca^2 依赖性蛋白激酶（PKC）抑制剂Bisindolylmaleimide Ⅰ、酷氨酸蛋白激酶（TPK）抑制剂Tyrphostin25分别预处理15min后,H2O2诱导的胞内活性氧的产生现象均消失;（4）细胞在无外钙环境下,小剂量H2O2诱导的胞内活性氧的产生明显减少;（5）细胞在无外钙环境下用NAC预处理后,H2O2诱导的胞内活性氧的产生现象消失。结果表明,H2O2可以通过胞内信号转导系统诱使WB细胞胞内活性氧产生,这可能与小剂量H2O2调控细胞生物学功能（如增殖、转化）相关。     

树木N2O排放速率的测定

 采用封闭罩法,对长白山阔叶红松林的几个主要树种——水曲柳 (Fraxinus mandshurica)、红松 (Pinus koraiensis)、毛赤杨（Alnus hirsuta）和椴树（Tilia amurensis）的连体枝叶排放N2O的速率进行了原位测定, 并考察了树木的N2O排放速率与土壤含水量的关系。结果表明：在观测期间,4种树木的N2O排放速率的变化范围分别为：水曲柳9.46～152.85 ng N2O·g－1 DW·h－1、红松1.37～64.51 ng N2O·g－1 DW·h－1、赤     

活性氧在外源乙烯诱导内源乙烯产生过程中的作用

外源乙烯在一定的条件下明显抑制了超氧化物歧化酶（SOD)和过氧化氢酶（CAT）的活性,提高了超氧化物阴离子自由基和过氧化氢（H2O2）的产率,从而有效地诱导了内源乙烯产生的增加;外源和H2O2对乙烯产生的促进作用及外源活性氧清除剂对乙烯产生的抑制作用也为此提供了证明。乙烯对植物生理过程的调节机制之一就是通过影响活性氧清除酶活性,从而调节各种活性氧在体内的平衡。     

大丽轮枝菌分泌蛋白激发子的分离纯化及生物功能研究

利用硫酸铵沉淀、(A)KTA explorer 10蛋白纯化仪、非变性电泳、割胶电洗脱等方法,从大丽轮枝菌(Verticillium dahliae)发酵液中分离纯化出一种蛋白激发子,经SDS-PAGE电泳检测单一条带,相对分子量为20 kD.该蛋白激发子能够诱导烟草的过敏反应,处理6h后,处理部位出现水溃状,24h后出现坏死斑.该激发子可以诱导烟草细胞在较短时间内产生防卫反应信号分子H2O2和NO,并引起活性氧爆发.     

低温诱导绿豆黄化幼苗乙烯产生过程中活性氧的作用

低温明显诱导绿豆 (PhaseolusradiatusL .)黄化幼苗乙烯产生速率的升高 ,同时也诱导活性氧产生速率不同程度的提高 ,显示二者之间有密切的关系。乙烯合成抑制剂AVG (2 aminoethoxyvinlglycine)、AOA(aminooxyaceticacid)能明显减弱低温对绿豆黄化幼苗乙烯产生的诱导作用 ,但对活性氧的产生没有明显的影响 ,说明低温诱导的乙烯产生的增加并不是活性氧产生增加的原因。超氧阴离子自由基 (O- ·2 )的特异性清除剂SOD和DABCO(1,4 diazabicyclo 2 ,2 ,2 octane)能有效削弱低温对乙烯产生的诱导作用 ,外源O- ·2 产生系统明显促进经过低温处理的幼苗回到常温下生长初期乙烯产生的增加 ,说明O- ·2 产生的增加可能是低温诱导乙烯产生增加的原因之一。低温诱导的H2 O2 产生的增加则被证明与乙烯产生速率的升高没有直接关系。     

新型真菌源激活蛋白诱导水稻抗病性及其生理机制

为明确新型真菌源激活蛋白对水稻抗病性的诱导作用及其生理机制,研究了激活蛋白对水稻稻瘟病和白叶枯病的诱导抗病性,监测了激活蛋白处理后水稻过氧化物酶（POD）、多酚氧化酶（PPO）、过氧化氢酶（CAT）、超氧化物歧化酶（SOD）活性及过氧化氢（H₂O₂）含量变化。结果表明,1～6 μg·mL^-1激活蛋白对稻瘟病和白叶枯病的诱抗效果分别为45.2%～71.4%和47.6%～66.3%,以6 μg·mL^-1激活蛋白的诱抗效果最好。与对照相比,2 μg·mL^-1激活蛋白处理水稻后3～15 d内不同程度诱导了防御酶POD、PPO和SOD活性,抑制CAT活性,提高H₂O₂含量。新型真菌源激活蛋白能够诱导水稻产生对稻瘟病和白叶枯病的抗病性,其诱导抗性机制与水稻体内的活性氧代谢密切相关。     

黄腐酸引发软骨细胞产生过氧化氢及硒的抑制作用

为探讨黄腐酸（FA）能否直接刺激软骨细胞产生活性氧以及硒能否抑制由此产生的活性氧,采用二氯荧光素双酯（DCFH－DA）作为软骨细胞产生过氧化氢的探针,用流式细胞技术定量地检测了在FA作用下软骨细胞产生的过氧化氢,并同时检测了硒存在时的过氧化氢含量．发现FA不但能够刺激软骨细胞产生过氧化氢（P＜0．05）,并与FA浓度相关,随FA浓度增大,产生过氧化氢增多,当FA浓度为100mg／L时软骨细胞产生过氧化氢达到最大,之后再增大FA浓度,过氧化氢的产生量减少;硒的存在对FA刺激软骨细胞产生过氧化氢有抑制作用．     

Hg2+胁迫对浮萍体细胞DNA一级结构和抗氧化酶系的损伤(英文)

 主要从DNA一级结构及抗氧化酶系变化两方面,研究了Hg2+胁迫下浮萍体细胞的损伤。结果表明：运用随机扩增多态性DNA法（Random amplified polymorphism DNA, RAPD）和DNA梯法（DNA Ladder）,5～10 mg·L-1　Hg2+处理组可检测到基因组DNA的明显损伤,20 mg·-1 Hg2+已导致细胞坏死;RAPD法较DNA Ladder法更灵敏。本文还发现,活性氧和抗氧化酶系很可能参与了浮萍体细胞凋亡过程。低浓度的Hg2+胁迫可刺激抗氧化酶活性升高,以清除体内活性氧,而一旦活性氧水平超出一定域值,抗氧化酶活性急速下降,导致细胞凋亡。      

Ca2+缓解 NaCl胁迫引起的玉米光合能力下降的作用

 以‘鲁玉14号玉米’（‘Zea mays cv. Luyu 14’）为材料,研究了Ca2+ 对NaCl胁迫下玉米叶片的气孔导度、净光合速率、Mehler反应及其对超氧化物歧化酶SOD和抗坏血酸过氧化物酶APX活性的影响,探讨了Ca2+在盐胁迫中的可能作用及其作用机制。正常生长条件下外施 2～16 mmol·L－1 Ca2+造成气孔导度下降,但在盐胁迫条件下外施 2～8 mmol·L－1 Ca2+却能降低盐胁迫造成的气孔导度下降。另外,无论正常生长条件下还是盐胁迫下,依赖Mehler反应的电子传递都会因 Ca2+的加入而增强,但后者的增强程度较大。对照植株经 2～8 mmol· L－1 Ca2+处理后总电子传递明显降低,而外施 2～8 mmol·L－1 Ca2+ 对盐胁迫植株的总电子传递却有明显的促进作用。8 mmol· L－1 Ca2+显著地提高了盐胁迫下SOD和APX的活性,相比之下,Ca2+对APX活性的促进作用更加显著。Ca2+的加入明显减轻了盐胁迫对玉米的抑制作用,这主要归因于Ca2+降低了盐胁迫造成的气孔关闭,改善了光合作用,并通过促进Mehler反应一方面直接耗散过剩的激发电子避免了电子传递链的过度还原,另一方面建立跨膜的pH梯度刺激依赖叶黄素循环的热耗散来耗散过剩光能,从而缓解了盐胁迫下过剩光能对玉米造成的伤害。而Mehler反应产生的活性氧可以被活性提高的抗氧化酶所清除。     

Photogeneration of singlet oxygen (1O2) and free radicals (Sen·-, O·-2) by tetra-brominated hypocrellin B derivative

To improve photodynamic activity of the parent hypocrellin B (HB), a tetra-brominated HB derivative (compound 1) was synthesized in high yield. Compared with HB, compound 1 has enhanced red absorption and high molar extinction coefficients. The photodynamic action of compound 1, especially the generation mechanism and efficiencies of active species (Sen^·-, O^·-₂ and ¹O₂) were studied using electron paramagnetic resonance (EPR) and spectrophotometric methods. In the deoxygenated DMSO solution of compound 1, the semiquinone anion radical of compound 1 is photogenerated via the self-electron transfer between the excited and ground state species. The presence of electron donor significantly promotes the reduction of compound 1. When oxygen is present, superoxide anion radical (O^·-₂) is formed via the electron transfer from Sens^·- to the ground state molecular oxygen. The efficiencies of Sens^·- and O^·-₂ generation by compound 1 are about three and two times as much as that of HB, respectively. Singlet oxygen (¹O₂) can be produced via the energy transfer from triplet compound 1 to ground state oxygen molecules. The quantum yield of singlet oxygen (¹O₂) is 0.54 in CHCl₃ similar to that of HB. Furthermore, it was found that the accumulation of Sens^·- would replace that of O^·-₂ or ¹O₂ with the depletion of oxygen in the sealed system.     

粤北石灰岩山地主要造林树种的生理生态学特性

本文是在调查粤北石灰岩山地植被的基础上,选出8种植物进行光能利用效率和蒸腾系数等生理生态学特性研究,并与非石灰岩山地较速生的荷木、藜蒴作比较,以筛选出适合石灰岩山地造林绿化的树种。结果表明,任豆、香椿与荷木、藜蒴的平均净光合速率(gCO2·m-2·d-1)分别为19.92,13.10,9.04和8.11;对光合有效辐射的利用效率(%)分别为4.21,4.59,2.71和2.52;蒸腾速率(gH2O·m-2·d-1)分别为2191,1266,1499和1133;蒸腾系数(gH2O·gDM-1)分别为164,144,244和209．说明任豆、香椿比荷木、藜蒴速生,光能利用效率、水分利用效率高,生态适应性强。光皮树、阴香的生长速度与荷木、藜蒴相似。菜豆树、桂花、铁榄、板栗的生长速度比荷木、藜蒴慢,但除板栗外都比较耐荫,对石灰岩山地环境也有较强的生态适应性。     

Characterization of 2,3-bis(chloromethyl)-1,4-naphthoquinone induced mitochondrial swelling

Mitochondrial swelling induced by 2,3-bis(chloromethyl)-1,4-naphthoquinone (CMNQ) was found to be a non-energy linked, oxygen and sulfhydryl-dependent, substrate-independent, osmotic process, that lacks cation specificity. Swelling was inhibited by cysteine and DTNB, and the CMNQ induced swelling resulted in a decrease in mitochondrial reactive sulfhydryl groups; thus, mitochondrial sulfhydryl interaction was mandatory in the CMNQ swelling process. The non-enzymatic reaction of CMNQ with cysteine but not cystine resulted in the consumption of oxygen, implicating sulfhydryl redox activity in the swelling process. High levels of tocopherol and histidine depressed the CMNQ induced swelling, suggesting that free radicals and singlet oxygen are important in the CMNQ induced swelling process.These findings support the proposition that CMNQ interacts with mitochondrial reductase systems and sulfhydryl groups in such a way as to generate superoxide radical which subsequently may dismute to H₂O₂ and produce ·OH and possibly singlet oxygen. These toxic oxygen species may be responsible for the CMNQ-promoted sulfhydryl depletion and mitochondrial swelling.     

Heat shock-induced attenuation of hydroxyl radical generation and mitochondrial aconitase activity in cardiac H9c2 cells

A mild heat shock (hyperthermia) protects cells from apoptotic and necrotic deaths by inducing overexpression of various heat shock proteins (Hsps). These proteins, in combination with the activation of the nitric oxide synthase (NOS) enzyme, play important roles in the protection of the myocardium against a variety of diseases. In the present work we report that the generation of potent reactive oxygen species (ROS), namely ·OH in cardiac H9c2 cells, is attenuated by heat shock treatment (2 h at 42°C). Western blot analyses showed that heat shock treatment induced overexpression of Hsp70, Hsp60, and Hsp25. The observed ·OH was found to be derived from the superoxide (O₂^–·) generated by the mitochondria. Whereas the manganese superoxide dismutase (MnSOD) activity was increased in the heat-shocked cells, the mitochondrial aconitase activity was reduced. The mechanism of O₂^–· conversion into ·OH in mitochondria is proposed as follows. The O₂^–· leaked from the electron transport chain, oxidatively damages the mitochondrial aconitase, releasing a free Fe²⁺. The aconitase-released Fe²⁺ combines with H₂O₂ to generate ·OH via a Fenton reaction and the oxidized Fe³⁺ recombines with the inactivated enzyme after being reduced to Fe²⁺ by other cellular reductants, turning it over to be active. However, in heat-shocked cells, because of higher MnSOD activity, the excess H₂O₂ causes irreversible damage to the mitochondrial aconitase enzyme, thus inhibiting its activity. In conclusion, we propose that attenuation of ·OH generation after heat shock treatment might play an important role in reducing the myocardial ischemic injury, observed in heat shock-treated animals. proteins; free radicals; spin trapping; reactive oxygen species     

SIMAZINE-INDUCED INHIBITION IN PHOTOACCLIMATED POPULATIONS OF ANABAENA CIRCINALIS (CYANOPHYTA)1

The effects of the triazine herbicide, simazine, on photosynthetic oxygen evolution and growth rate in photoacclimated populations of Anabaena circinalis Rabenhorst were investigated. Chemostat populations were acclimated to photon flux densities (PFDs) of 50, 130, and 230 μmol·m^?2·s^?1 of photosynthetic active radiation (PAR), Decreases in chlorophyll a (Chl a). c-phycocyanin (CPC), and total carotenoid (TCar) contents and CPC: Chl a and CPC: TCar ratios of populations coincided with increasing PFD, Polynomial regression models that characterize inhibition of photosynthesis for populations acclimated to 50 and 130 μmol photons·m^?2·s^?1 PAR were distinct from the model for populations acclimated to 230 μmol photons·m^?2·s^?1 PAR. Simazine concentrations that, depressed oxygen evolution 50% compared to controls decreased with increasing PFD. Increases and decreases in both biomass and growth rate coincided with increasing PFD and simazine concentration, respectively. Simazine concentrations that depressed growth rate 50% compared to controls increased with decreasing PFD. The differences in photosynthetic and growth inhibition among photoacclimated populations indicate that sensitivity to photosystem II inhibitors is affected by alterations in pigment contents.     

NNK诱发BEP2D细胞产生活性氧及其对DNA的损伤

通过测定细胞内和细胞上清中活性氧（ｒｅａｃｔｉｖｅ ｏｘｙｇｅｎ ｓｐｅｃｉｅｓ,ＲＯＳ）水平,以及ＤＮＡ 加合物——８－羟基脱氧鸟嘌呤核苷（８－ｈｙｄｒｏｘｙｄｅｏｘｙｇｕａｎｏｓｉｎｅ,ＯＨ８ｄＧ）含量,对烟草特异亚硝胺类化合物４－甲基亚硝胺－１（３－吡啶基）－１－丁酮（４－（ｍ ｅｔｈｙｌｎｉｔｒｏｓａｍ ｉｎｏ）－１－（３－ｐｙｒｉｄｙｌ）－１－ｂｕｔａｎｏｎｅ,ＮＮＫ）诱发人乳头状病毒永生化的人支气管上皮细胞（ｈｕｍ ａｎ ｐａｐｉｌｌｏｍ ａｖｉｒｕｓ－ｉｍ ｍ ｏｒｔａｌｉｚｅｄ ｈｕｍ ａｎｂｒｏｎｃｈｉａｌｅｐｉｔｈｅｌｉａｌｃｅｌｌｌｉｎｅ,ＢＥＰ２Ｄ）产生的ＲＯＳ及其对ＤＮＡ 的氧化损伤进行研究,并观察纳米硒的保护作用．结果表明,ＢＥＰ２Ｄ 细胞经不同浓度的ＮＮＫ 作用后,细胞内和细胞上清中ＲＯＳ以及ＯＨ８ｄＧ含量均显著增加,并有较好的剂量效应关系．１ μｍ ｏｌ·Ｌ－ １纳米硒（ｎａｎｏｓｅｌｅｎｕｉｍ ,ＮＳ）能明显抑制ＮＮＫ 诱发ＢＥＰ２Ｄ细胞产生的ＲＯＳ及ＯＨ８ｄＧ 水平．揭示ＮＮＫ 能造成细胞的氧化损伤,而ＮＳ对ＮＮＫ 所致细胞的氧化损伤有保护作用．     

Chemiluminescence study of active oxygen species produced by TiO2 photocatalytic reaction.

Two chemiluminescence approaches have been used for study of active oxygen species produced by the TiO2 photocatalytic reaction. One is based on flow injection analysis (FIA)-luminol chemiluminescence (CL); another is a time-resolved CL method. In the FIA-CL experiment, an UV-illuminated TiO2 suspension and water were passed into a mixing cell by two separate flow lines. Luminol solution was injected into the water flow line at different times. The injected luminol reacted with active oxygen species generated by the TiO2 photocatalytic reaction in a mixing coil and produced CL. It was found that the maximum CL was detected at the first injection of luminol. CL intensity decreased with time of injection. When the luminol was injected after 5 min, the CL intensity was almost unchanged. Addition of scavengers of active oxygen species indicated that the CL produced early in the 5 min was caused by O2- and H2O2, while CL after 5 min was only from H2O2. In the time-resolved CL, the third harmonic wavelength of Nd:YAG laser (355 nm) was used as a UV light source, and CL was detected by a PMT and recorded in a millisecond time scale using a digital oscilloscope. It was found that CL induced by the photocatalytic reaction increased with concentration of the TiO2 suspension. Scavengers of active oxygen species of *OH, O2- and H2O2 were added to study the involvement of the active oxygen species.     

Paramagnetic Induced Shifts in the Proton Magnetic Resonance Spectra of Cyclopropanecarboxylic Acid Esters Using Eu(fod)3

The generation of active oxygen molecules, O₂^?, H₂O₂, and · OH, from the aqueous solution of aminosugars, such as d-glucosamine, was confirmed by their actual measurement. Both the C-2 amino and C-1 aldehyde groups in the aminosugar molecules were indispensable for the generation of active oxygen molecules. The introduction of a C-6 phosphate group to d-glucosamine or the simultaneous use of phosphate ion and d-glucosamine heightened the original activity of d-glucosamine to generate these oxygens, especially · OH. Cu²⁺, which promoted the DNA-breaking activity of aminosugar most at 1 mm, also promoted the generation of · OH most greatly at the same concentration, but neither O₂^? nor H₂O₂ was generated under the same conditions. Superoxide dismutase, catalase, and some radical scavengers inhibited the generation of these active oxygen molecules. Among the active oxygen molecules, only the amount of · OH generated was directly proportional to the DNA-breaking activity of the aminosugar.