In vivo studies on the toxic effects of microcystins on mitochondrial electron transport chain and ion regulation in liver and heart of rabbit

This study examined the toxic effects of microcystins on mitochondria of liver and heart of rabbit in vivo. Rabbits were injected i.p. with extracted microcystins (mainly MC-RR and -LR) at two doses, 12.5 and 50 MCLReq. mu g/kg bw, and the changes in mitochondria of liver and heart were studied at 1, 3,12, 24 and 48 h after injection. MCs induced damage of mitochondrial morphology and lipid peroxidation in both liver and heart. MCs influenced respiratory activity through inhibiting NADH dehydrogenase and enhancing succinate dehydrogenase (SDH). MCs altered Na+-K+-ATPase and Ca2+-Mg2+-ATPase activities of mitochondria and consequently disrupted ionic homeostasis, which might be partly responsible for the loss of mitochondrial membrane potential (MMP). MCs were highly toxic to mitochondria with more serious damage in liver than in heart. Damage of mitochondria showed reduction at 48 h in the low dose group, suggesting that the low dose of MCs might have stimulated a compensatory response in the rabbits. (C) 2008 Elsevier Inc. All rights reserved.

Effects of La3+ on lipid fluidity land structural transitions in human erythrocyte membranes

The effects of La3+ on the structure and function of human erythrocyte membranes were investigated by fluorescence polarization, spin-labeled electron spin resonance (ESR) and differential scanning calorimetry (DSC). The results showed that increasing concentrations of La3+ inhibited (Na++K+)-ATPase and Mg2+-ATPase activities. La3+ lowered the lipid fluidity of erythrocyte membranes and induced structural transitions in erythrocyte membranes.

INTERACTION OF LANTHANUM AND CALCIUM WITH HUMAN ERYTHROCYTE-MEMBRANES

The effect of lanthanum and calcium on the structure and function of human erythrocyte membranes was investigated by fluorescence polarization, spin- labeled electron spin resonance (ESR) and laser Raman spectroscopy. The results showed that low concentration of La3+ (0.5 mu mol/L) activated a Little (Na++K+)-ATPase and Mg2+-ATPase activities, and it inhibited obvi ously the ATPase activities with increasing its concentrations. La3+ lowered the lipid fluidity of human erythrocyte membranes and decreased the vibration intensity of alpha-helix of the protein in the Amide I '. The effect of Ca2+ on the lipid fluidity and alpha-helix of the protein in the Amide I ' was smaller than that of La3+.

塔玛亚历山大藻(Alexandrium tamarense)对鲈鱼(Lateolabrax japonicus) 的危害机制研究

本论文选择在我国分离得到的一株有毒赤潮甲藻-塔玛亚历山大藻（Alexandrium tamarense，ATHK株），研究了其对一种我国沿海常见和典型养殖鱼类鲈鱼（Lateolabrax japonicus）的危害机制。首先研究了塔玛亚历山大藻（ATHK）对鲈鱼鳃结构的影响及其溶血毒性； 然后采用腹腔注射的方法，研究了高剂量塔玛亚历山大藻毒素（ATHK毒素：约为1.6×105 细胞，相应PSP为0.886µg STX Equal，相当于每克湿重的鲈鱼PSP注射量为0.0118µg STX Equal）和低剂量塔玛亚历山大藻毒素（ATHK毒素： 约为0.16×105 细胞，相应PSP为0.0886µg STX Equal，相当于每克湿重的鲈鱼PSP注射量为0.00118µg STX Equal）在鲈鱼体内代谢过程中对鲈鱼肝脏、肾脏和鳃组织的超微结构、Na+K+-ATPase活性、肝脏功能和肾脏功能的影响、以及对抗氧化系统酶活性和异生物质代谢酶的影响，以期从不同方面了解塔玛亚历山大藻及其所产水溶性毒素（ATHK毒素）对鲈鱼的毒害效应及机制，为有毒赤潮的有效管理提供一定的科学依据。 塔玛亚历山大藻（ATHK）对鲈鱼鳃组织影响的实验结果表明，该藻使鲈鱼鳃组织出现水肿现象，细胞间隙变大；粘液细胞颗粒不规则，颜色加深，颗粒发生凝集，有板结状；氯细胞线粒体内部基质凝集。不产PSP的一种亚历山大藻（AT-6）也使鲈鱼鳃出现水肿，且使细胞出现一定的固缩现象。显微镜观察发现鲈鱼鳃丝间存在有这两种亚历山大藻细胞。由此推测塔玛亚历山大藻ATHK和AT-6的表面结构可能具有能导致鲈鱼鳃组织水肿的机械作用。对人血细胞溶血实验结果表明塔玛亚历山大藻（ATHK）具有较强的溶血毒性，大小与藻的生长阶段和细胞密度都有一定的关系：指数期的溶血毒性最大，随细胞数目的增多，活性逐渐加大；藻细胞、细胞碎片、细胞内容物都有一定的溶血毒性，其中细胞碎片的活性最大。通过11种（株）产PSP的亚历山大藻、不产PSP的亚历山大藻以及标准PSP的实验结果表明这种溶血毒性是由藻细胞的其它非PSP物质造成的，且这种溶血毒性在产PSP的亚历山大藻中具有一定的普遍性。 塔玛亚历山大藻（ATHK）对鲈鱼组织超微结构实验结果表明：ATHK毒素（约为1.6×105 细胞，相应PSP为0.886µg STX Equal，相当于每克湿重的鲈鱼PSP注射量为0.0118µg STX Equal）能导致鲈鱼组织细胞超微结构发生剧烈的变化，主要表现在：肝细胞细胞膜有肿胀现象，部分膜边缘溶解；细胞质糖原颗粒化；核糖体脱落，仅见滑面内质网；细胞质和线粒体内都出现空泡，且线粒体的嵴状结构也发生变化；核膜溶解比较严重，核质外溢，且异染色质边际化。前肾细胞超微结构的变化主要是淋巴细胞核质出现空泡，核膜有溶解迹象；Ⅰ型粒细胞颗粒膨大，伪足增多且变长；Ⅱ型颗粒细胞颗粒增多，内部出现空腔，细胞膜和核膜溶解，胞质、细胞器和核质外溢。鳃组织中氯细胞的核膜局部溶解，核仁弥散，线粒体膜溶解，微细小管膨大；粘液颗粒膜溶解，内部结构遭受破坏；扁平细胞核膜及线粒体膜几乎全部溶解。因此，我们的结果表明，ATHK毒素能作用于鲈鱼细胞的内膜和外膜系统，使膜发生溶解、脱落等变化；比较注射同样剂量大小ATHK毒素120h和240h时鲈鱼组织超微结构发现，细胞超微结构在一定程度上能够恢复。 ATHK毒素对Na+K+-ATPase活性、肝脏功能以及肾脏功能的影响结果表明，0.16×105―1.6×105细胞范围内的ATHK毒素可以显著影响肝脏和鳃组织中的Na+K+-ATPase，使这两种组织中的Na+K+-ATPase活性出现不同程度的下降； 而且还能够显著抑制肝脏中谷丙转氨酶的活性，最大抑制率为95%。但此范围内的ATHK毒素不能显著影响肾脏中的Na+K+-ATPase活性以及尿素氮含量。因此，ATHK毒素对Na+K+-ATPase活性的抑制则会导致细胞能量的缺失，使细胞进一步发生其它变化，而ATHK毒素对肝脏功能完整性的影响则可能会抑制对蛋白质的分解代谢。 ATHK毒素对鲈鱼肝脏、肾脏和鳃组织中的、超氧化物歧化酶（SOD）、过氧化氢酶（CAT）、谷胱肝肽过氧化物酶（GSH-Px）以及谷胱肝肽转硫酶（GST）活性变化影响的结果表明：高剂量（约为1.6×105 细胞，相应PSP为0.886µg STX Equal，相当于每克湿重的鲈鱼PSP注射量为0.0118µg STX Equal）ATHK毒素能显著诱导鲈鱼肝脏和鳃组织中SOD、GSH-Px以及GST酶活性，最大变化范围为正常状态下的3-4倍，对肝脏中CAT酶活性具有一定的抑制作用，对鳃中的CAT抑制效应则不显著；但此剂量的ATHK毒素仅对肾脏鳃中的GSH-Px活性有一定的诱导作用，对SOD、CAT以及GST的活性没有显著影响。低剂量（约为0.16×105 细胞，相应PSP为0.0886µg STX Equal，相当于每克湿重的鲈鱼PSP注射量为0.00118µg STX Equal）ATHK毒素也能诱导鲈鱼肝脏和鳃组织中SOD、GSH-Px以及GST酶活性，其在第一个24h内的诱导效果与临界致死毒素剂量相似； 且连续注射低剂量ATHK毒素则对肝脏和鳃中这三种酶活性具有累加的诱导作用，使这三种酶活性的变化范围为正常的5倍；低剂量ATHK毒素对肝脏中CAT酶活性也具有抑制作用，但对鳃中CAT酶活性的抑制作用并不显著。 同样，低剂量ATHK毒素除对肾脏中GSH-Px活性具有一定诱导效应外，对SOD、CAT以及GST都没有显著影响。 SOD、GSH-Px以及GST酶活性的显著升高表明ATHK毒素在鲈鱼体内代谢过程中能诱导鲈鱼产生活性氧自由基，且GST活性的升高则说明作为细胞色素P450依赖的异生物质代谢酶，GST在ATHK毒素代谢过程中可能可以加速ATHK毒素的代谢。推测鲈鱼可以通过这三种酶降低ATHK毒素以及次生毒物活性氧自由基对鲈鱼的危害。

两种海藻提取物的化学成分和生物活性的研究及其应用

本论文的研究工作由两部分组成，第一部分研究了海带（Laminaria japonica）水提取物中的活性物质，并研究了提取物对蔬菜促生长的影响及其作用机制。第二部分对三列凹顶藻(Laurencia tristicha)乙醇提取物的乙酸乙酯相进行了活性筛选和化学成分研究，并对其中分离得到的单体化合物进行了生物活性筛选。 第一部分主要以中国人工养殖的海带为原料，使用与海藻多糖生产相结合的提取技术并浓缩其中的有效成分。对浓缩提取物进行了蔬菜的农田效果实验，并对作物抗旱性能的增加、作物硝酸盐积累的减少、作物品质的改善、以及作物抵抗病毒病的能力等影响进行了作用机制方面的研究。海藻浓缩提取物进行的农田效果实验表明：作物抗旱型相对含水量RWC值在92%~94%之间；病毒病的防治效果最高可达到91%；作物的品质有明显的改善，最重要的是首次发现海藻提取物有降低蔬菜中硝酸盐的含量（硝酸盐的含量是与有机蔬菜区别的重要指标之一）的作用。该部分研究工作的创新性主要体现在：（1）首次在国内外提出和采用与海藻多糖生产相结合的提取技术。该技术的应用不但减少了提取成本，使工业化生产成为可能,更重要的是使我国的海藻工业生产可能实现高值化和开辟综合利用的新途径。（2）首次发现海藻中的小分子海藻多糖具有和细胞激动素、甜菜碱、植物生长素等活性物质同样的生物活性。 第二部分的研究是在查阅了大量的近20年来国内外有关红藻凹顶藻中化学成分研究的相关文献的基础上，对凹顶藻中的次生代谢产物进行了综述。该论文主要是通过对红藻三列凹顶藻的95%乙醇提取物的乙酸乙酯相进行化学成分分析和生物活性筛选以期能够发现具有药用前景的活性先导化合物。 为了寻找具有生物活性的化合物，我们对采自我国南海硇洲岛海域的红藻三列凹顶藻的95%乙醇提取物的乙酸乙酯相进行了活性筛选。采用MTT法对其在KB细胞株、Bel-7402细胞株、PC-3M细胞株、MCF-7细胞株、Ketr-3细胞株模型上进行了细胞毒活性测试；采用酶模型对其进行了Na+,K+-ATPase的抑制活性测试；采用MTT法对其在犬主动脉血管模型上进行了血管平滑肌细胞增殖抑制活性测试；结果表明，三列凹顶藻的95%乙醇提取物的乙酸乙酯相对Na+,K+-ATPase和犬血管平滑肌细胞增殖具有一定的抑制活性。 利用正相和反相色谱、Sephadex LH-20色谱以及反相HPLC等手段进行分离纯化，从我国南海海域的红藻三列凹顶藻中分离得到33种化学成分，通过波谱学方法（IR、MS、NMR）以及X-ray单晶衍射试验对其化学结构进行了确证，其中化合物L1~L8为新结构化合物，化合物L5为具有新骨架的全新结构化合物，化合物L9~L13为新天然产物，化合物L18和L22系首次从海洋生物中获得，所有化合物均为首次从该属海藻中得到。新化合物L1~L8均为倍半萜类化合物，命名分别为：(1R,3R)-(-)-3-(3-hydroxy-4-methylphenyl)- 1,3-dimethyl–2-methylidene cyclopentanol (L1), (1R,3R)-(-)-3-(4-methylphenyl)-1,3-dimethyl-2-methylidenecyclopentanol (L2)， (1R, 3R)-(-)-3-(2-hydroxy-4-methylphenyl)-1,3-dimethyl–2–methylidenecyclopentanol (L3)，(+)-(1S,2R)–2-(3–hydroxy–4–methylphenyl)-1,2-[3.1.0]bicy-clohexane (L4)，()-(1S,2R) -5-hydroxy–6–methyl-spiro-dihydrobenzofuran-2(3H),2-{1-methyl-[3.1.0]bicyclohexane} (L5), (+)-6-methyl-2-(p-tolyl)hept-4-en-2,6-diol (L6)，(3R,3aS,8bS)-(-)-2,3,3a,8b–tetrahydro–7-bromo – 3 a– hydroxymethyl - 3, 6, 8b - trimethyl-1H- cyclopenta[b] benzofuran (L7 )，(3R, 3aS, 8bS) - (-) - 2,3,3a,8b–tetrahydro–3 a–hydroxymethyl-3,6,8b -trimethyl -1H – cyclopenta [b] benzofuran (L8)。25个已知结构化合物确定为：(+)-(1R,2R)-4-bromo-1,5, 9–trimethyl–12– methylidene–8–oxa-tricyclo[7.2.1.02]dodeca-2,4,6-triene (L9)，(3S,3aR,8bS)-(-)-2,3,3a, 8b– tetra -hydro–7-bromo–3–hydroxy-3,3a,6,8b-tetramethyl-1H-cyclopenta[b]benzofu- ran (L10 )，(3R, 3aR, 8bS) - (-) - 2, 3, 3a, 8b – tetrahydro – 7 - bromo – 3 – hydroxy - 3,3a,6,8b - tetramethyl - 1H - cyclopenta [b] benzofuran (L11 )，(3S,3aR,8bS) - (-) - 2, 3, 3a, 8b – tetrahydro –3–hydroxy -3, 3a, 6, 8b - tetramethyl-1H-cyclopenta[b]benzofuran (L12 )， ( 3aR, 8bS) - (-) - 3a,8b –dihydro–7 - bromo – 3, 3a, 6, 8b - tetramethyl - 1H - cyclopenta[b]benzofuran (L13 )，aplysinol (L14 ) ，aplysin (L15)，laurebiphenyl (L16)，johnstonol (L17)，gossonorol (L18)，7,10-epoxy-ar- bisabol-11-ol (L19)，10-epi-7,10-epoxyarbisabol-11-ol (L20) 3β-hydroxy- 5α, 6α-epoxy- β- ionone (L21 )，3β-hydroxy-5β,6β-epoxy-β-ionone (L22 )，胆甾醇 (L23 )，胆甾-5-烯-3β,7α二醇胆甾-5-烯-3β,7α二醇 (L24)，β-谷甾醇 (L25)，叶绿醇 (L26 )，玉米黄素 (L27 )，对羟基苯甲醛 (L28 )，3-吲哚甲醛 (L29 )，1-O-十六烷酰基-3-O-β-D-吡喃半乳糖基-丙三醇(L30 )，1-O-十八烷酰基-3-O-β-D-吡喃半乳糖基-丙三醇 (L31 )，丙三醇-1-软脂酸单酯 (L32 )，正十六碳酸 (L33 )。 采用MTT法对其中23个单体化合物在Bel-7402细胞株、BGC-823细胞株、A549细胞株、A2780细胞株、HCT-8细胞株和HELL细胞株模型上进行了细胞毒活性测试；采用MTT法对其中13个单体化合物在犬主动脉血管模型上进行了血管平滑肌细胞增殖抑制活性测试；结果表明，部分单体化合物显示出一定的生物活性。

微生物——铊相互作用的生物地球化学研究——以真菌（Fungus）为例

铊是一种有毒有害的重金属元素，已经引起了广泛的关注。本论文通过对黔西南铊矿区土壤和沉积物样品的菌株分离、铊高耐受性菌株的筛选、胞外吸附、富集、亚细胞水平区系分布、絮凝实验及ITS序列等实验研究分析，并结合铊的地球化学相关研究，较系统地阐述了真菌--铊的生物地球化学过程机理，得出以下结论： 1、与环境背景区相比，黔西南滥木厂铊矿区内的河流、土壤中铊的已有不同程度的积累，直接导致了当地微生物生物量在很大程度上的降低，微生物生物量与铊含量间有显著的负相关关系。研究区内的沉积物、土壤中的微生物区系结构和数量发生了明显变化，细菌、真菌及放线菌数量均出现显著降低，而且三大微生物对重金属污染的敏感性大小也不一样，即放线菌>细菌>真菌。从土壤样品中分离到的主要菌群仍为常见种属，如青霉属（Penicillium）、木霉属（Trichoderma）、拟青霉（Paecilomyces）等。 2、经过初筛菌株的铊耐受性实验，在1000 mg/L水平筛选得到九株高耐受性菌株。吸附实验表明：微生物菌株对铊的吸附效率在4.63～16.89%，且随着环境中铊浓度的上升而降低，这可能是因为铊浓度的升高加大了对微生物生长的抑制作用，所形成的菌丝体（或菌丝球）减少，表面积也相应减少，从而导致了吸附效率的下降。各种常量元素和铊的关系呈显著相关性，钙、钾和钠等常量元素也是微生物赖以维持生存的因子，可能由于微生物细胞对钙、钾的吸附方式与对铊的吸附方式类似。因此，随着铊处理浓度的上升，钙和钾的吸附量也随之减少，而钠则呈现相反的趋势。 3、富集实验表明，九株菌株对铊的富集量随着铊处理浓度上升而降低，其影响趋势与对生物量的影响趋势基本一致，最高可达到7189 mg/kg，最大富集系数为7.2。九株菌株对常量元素的富集与对铊的富集并无明显的相关性，但在考察铊处理浓度对常量元素的富集影响时发现，铊处理浓度的上升与对钙的富集量表现出较强的正相关；而对钾、钠、镁的富集影响并不明显。 4、亚细胞水平上的铊分布研究表明，铊的富集优先顺序为：细胞质>细胞壁>细胞器。亚细胞水平的区隔化作用是微生物对铊的主要耐受机制，细胞质是赋存铊的主要场所（53.83～79.45 %）。结合各亚细胞组分中常量元素与铊之间的相关性，并联系前人的研究，Tl+主要是通过细胞壁的Na+ -K+ ATPase和K+ -电位门通道进入细胞内的从而影响细胞的正常代谢的，而Ca2+的活化更有助于这一过程。 5、絮凝实验表明，培养三天后的发酵液对矿区废水中铊的去除率最高可达到70.49 %，最佳影响因子组合为：pH=8，温度为16℃，搅拌时间为4分钟。菌株的絮凝活性最高可达到57.32%，最佳影响因子组合为：pH=8，温度为14℃，搅拌时间为4分钟。 6、通过对九株铊高耐受性菌株的ITS序列分析及其在Gene Bank中的BLAST比对结果表明，五株菌株同属于木霉属（Trichoderma），两株菌株同属于青霉属（Penicillium）。这表明这两类真菌对铊的适应性较强，为以后寻找铊高耐受性菌株及其资源化利用提供了理论基础。

Biological application of multi-component nanowires in hybrid devices powered by F-1-ATPase motors

In this paper, construction of hybrid device by integrating nanowires with F1-ATPase motors is described. The nickel nanowires and multi-segment nanowires, including gold and nickel, were fabricated by electrochemical deposition in nanoporous templates. The nickel nanowires functionalized by biotinylated peptide can be assembled directly onto F1-ATPase motors to act as the propellers. If the multicomponent nanowires, including gold and nickel, were selectively functionalized by the thiol group modified ssDNA and the synthetic peptide, respectively, the biotinylated F1- ATPase motors can be attached to the biotinylated peptide on nickel segment of the nanowires. Then, the multi-component nanowires can also be used as the propellers, and one may observe the rotations of the multi-component nanowires driven by F1-ATPase motors. Therefore, introduction of multiple segments along the length of a nanowire can lead to a variety of multiple chemical functionalities, which can be selectively bound to cells and special biomolecules. This method provides an insight for the construction of other hybrid devices with its controlling arrangement of different biomolecule on designed nanometer scale structures.

Direct numerical test of the B-G-K model equation by the DSMC method

In the present paper the rarefied gas how caused by the sudden change of the wall temperature and the Rayleigh problem are simulated by the DSMC method which has been validated by experiments both in global flour field and velocity distribution function level. The comparison of the simulated results with the accurate numerical solutions of the B-G-K model equation shows that near equilibrium the BG-K equation with corrected collision frequency can give accurate result but as farther away from equilibrium the B-G-K equation is not accurate. This is for the first time that the error caused by the B-G-K model equation has been revealed.

用自相关法确定VITA法的门限K

研究了用ＶＩＴＡ条件采样方法壁湍猝发的门限值与壁湍流猝发平均周期之间的关系，根据用壁湍流流向脉动速度自相关函数检测得到的壁湍流猝发平均周期，确定壁，湍猝发ＶＩＴＡ条件采样的门限值Ｋ。

Asymptotic analysis based on K-S constitutive law for a rubber wedge compressed by a line load at its tip

In the present paper, a rubber wedge compressed by a line load at its tip is asymptotically analyzed using a special constitutive law proposed by Knowles and Sternberg (K-S elastic law) [J. Elasticity 3 (1973) 67]. The method of dividing sectors proposed by Gao [Theoret. Appl. Fract, Mech. 14 (1990) 219] is used. Domain near the wedge tip can be divided into one expanding sector and two narrowing sectors. Asymptotic equations of the strain-stress field near the wedge tip are derived and solved numerically. The deformation pattern near a wedge tip is completely revealed. A special case. i.e. a half space compressed by a line load is solved while the wedge angle is pi.

Properties of high k gate dielectric gadolinium oxide deposited on Si(100) by dual ion beam deposition (DIBD)

Gadolinium oxide thin films have been prepared on silicon (100) substrates with a low-energy dual ion-beam epitaxial technique. Substrate temperature was an important factor to affect the crystal structures and textures in an ion energy range of 100-500 eV. The films had a monoclinic Gd2O3 structure with preferred orientation ((4) over bar 02) at low substrate temperatures. When the substrate temperature was increased, the orientation turned to (202), and finally, the cubic structure appeared at the substrate temperature of 700 degreesC, which disagreed with the previous report because of the ion energy. The AES studies found that Gadolinium oxide shared Gd2O3 structures, although there were a lot of oxygen deficiencies in the films, and the XPS results confirmed this. AFM was also used to investigate the surface images of the samples. Finally, the electrical properties were presented. (C) 2004 Elsevier B.V. All rights reserved.

Starting Nozzle Flow Simulation Using K-G Two-Equation Turbulence Model

The starting process of two-dimensional nozzle flows has been simulated with Euler, laminar and k - g two-equation turbulence Navier-Stokes equations. The flow solver is based on a combination of LUSGS subiteration implicit method and five spatial discretized schemes, which are Roe, HLLE, MHLLE upwind schemes and AUSM+, AUSMPW schemes. In the paper, special attention is for the flow differences of the nozzle starting process obtained from different governing equations and different schemes. Two nozzle flows, previously investigated experimentally and numerically by other researchers, are chosen as our examples. The calculated results indicate the carbuncle phenomenon and unphysical oscillations appear more or less near a wall or behind strong shock wave except using HLLE scheme, and these unphysical phenomena become more seriously with the increase of Mach number. Comparing the turbulence calculation, inviscid solution cannot simulate the wall flow separation and the laminar solution shows some different flow characteristics in the regions of flow separation and near wall.