大黄鱼过氧化物酶Ⅳ在人胚肾细胞中的表达与抗氧化活性测定

为了研究大黄鱼（Pseudosciana crocea）peroxiredoxin Ⅳ（Lyc-Prx Ⅳ）在细胞内的抗氧化功能,构建了表达大黄鱼Prx Ⅳ的重组质粒p CMV-Lyc-Prx Ⅳ,瞬时转染人胚肾细胞（HEK-293T）,利用Western blotting方法检测转染后的细胞样品中Lyc-Prx Ⅳ的表达情况,并通过测定细胞内过氧化氢浓度来评价Lyc-Prx IV的体内抗氧化作用.结果显示,Lyc-Prx Ⅳ可在转染重组质粒p CMV-Lyc-Prx Ⅳ的HEK-293T细胞中表达,且细胞中A560处的吸光值在转染后6、12、24 h后分别为0.154、0.116以及0.162,而瞬时转染空载体p CMV后细胞样品在A560处的吸光值在转染后一直稳定在0.260左右,说明细胞中过氧化氢的浓度在转染后6、12、24 h时明显低于瞬时转染空载体p CMV细胞中的过氧化氢浓度（p〈0.01）.表明Lyc-Prx Ⅳ在生物体内可以分解过氧化氢,参与生物体内氧化还原状况的调控.     

Production of dimethylsulfide and acrylic acid from dimethylsulfoniopropionate during growth of three marine microalgae

We measured the concentrations of dimethylsulfide （DMS）, acrylic acid （AA）, and dimethylsulfoniopropionate （DMSP） during growth of three microalgae： Prorocentrum micans, Gephyrocapsa oceanica, and Platymonas subcordiformis. The DMSP, AA, and DMS concentrations in culture media varied significantly among algal growth stages, with the highest concentrations in the late stationary growth stage or the senescent stage. In the stationary growth stage, the average DMSP concentration per cell in P. micans （0.066 5 pmol/cell） was 1.3 times that in G. oceanica （0.049 5 pmol/ cell） and 20.2 times that in P. subcordiformis （0.003 29 pmol/cell）. The average concentrations of AA were 0.044 6, 0.026 9, and 0.003 05 pmol/cell in P. micans, G. oceanica, and P. subcordiformis, respectively, higher than the concentrations of DMS （0.272, 0.497, and 0.086 2 fmol/cell, respectively）. There were significant positive correlations between cell density and AA, DMSP, and DMS concentrations. The ratios of DMS/AA and AA/（DMSP＋AA） in the three algae differed significantly over the growth cycle. In all three microalgae, the DMS/AA ratios were less than 25% during the growth period, suggesting that the enzymatic cleavage pathway, which generates DMS, was not the main DMSP degradation pathway. The changes in the DMS/AA ratio indicated that there was a higher rate of enzymatic breakdown of DMSP in the early growth period and a lower rate during senescence. In all three microalgae, the AA/（DMSP＋AA） ratio （degradation ratio of DMSP） decreased during the exponential growth phase, and then increased. The variations in these ratios can approximately indicate the cleavage mechanism of DMSP at different stages of algal growth.     

Effects of rice straw on the cell viability,photosynthesis, and growth of Microcystis aeruginosa

Rice straw is supposed to be an environment-friendly biomaterial for inhibiting the growth of harmful blooms of the cyanobacterium Microcystis aeruginosa. However, its potential mechanism is not well known. To explore this mechanism, the growth, cell viability （esterase activity, membrane potential, and membrane integrity）, photosynthesis, and cell size ofM. aeruginosa were determined using flow cytometry and Phyto-PAM after exposure to rice straw extracts （RSE）. The results show that doses from 2.0 to 10.0 g/L of RSE efficiently inhibited the alga for 15 days, while the physiologic and morphologic responses of the cyanobacteria were time-dependent. RSE interfered with the cell membrane potential, cell size, and in vivo chlorophyll-a fluorescence on the first day. After 7 days of exposure, RSE was transported into the cytosol, which disrupted enzyme activity and photosynthesis. The cyanobacteria then started to repair its physiology （enzyme activity, photosynthesis） and remained viable, suggesting that rice straw act as an algistatic agent.     

Preparation,Characterization and Pharmacokinetics of Fluorescence Labeled Propylene Glycol Alginate Sodium Sulfate

A rapid and sensitive fluorescence labeling method was developed and validated for the microanalysis of a sulfated polysaccharide drug,namely propylene glycol alginate sodium sulfate （PSS）,in rat plasma.Fluorescein isothiocyanate （FITC） was selected to label PSS,and 1,6-diaminohexane was used to link PSS and FITC in order to prepare FITC-labeled PSS （F-PSS） through a reductive amination reaction.F-PSS was identified by UV-Vis,FT-IR and ^1H-NMR spectrum.The cell stability and cytotoxicity of F-PSS were tested in Madin-Darby canine kidney （MDCK） cells.The results indicated that the labeling efficiency of F-PSS was 0.522%±0.0248% and the absolute bioavailability was 8.39%.F-PSS was stable in MDCK cells without obvious cytotoxicity.The method was sensitive and reliable; it showed a good linearity,precision,recovery and stability.The FITC labeling method can be applied to investigating the absorption and metabolism of PSS and other polysaccharides in biological samples.     

褐潮藻Aureococcus anophagefferens的危害研究进展

<正>褐潮(Brown tide)是一种有害藻华,目前已知的主要藻种为Aureococcus anophagefferens及Aureoumbra lagunensis,由于藻华期间海水呈褐色,因此被称为褐潮[1]。褐潮的爆发严重危害到海洋资源和贝类养殖业,并且影响到海洋生物的种群结构乃至整个海洋生态系统。在美国东海岸,褐潮导致当地海湾扇贝资源崩溃、硬壳蛤资源衰退和海草床生境退化等[2];     

卡罗藻毒素研究进展

<正>剧毒卡罗藻Karlodinum veneficum(又称Karlodinium micrum)作为一种典型的赤潮种,是海水养殖水域和自然水体中一种常见的裸甲藻种类,为混合营养型生物。该藻自20世纪50年代被发现以来,在过去的60年中其命名曾发生过多次变更,直到最近将其归入裸甲藻种类,被译为剧毒卡罗藻[1](也有学者命名为剧毒卡尔藻[2])。该藻自首次在南非被发现     

细胞小片处理对马氏珠母贝育珠效果影响

以马氏珠母贝选系F2为实验材料,按照马氏珠母贝常规技术进行插核手术,并经过池塘休养和海区养殖过程,比较了细胞小片黏液处理和保养对马氏珠母贝育珠效果影响.实验设为4组:细胞小片黏液处理组(A)、细胞小片保养组(B)、同时进行细胞小片黏液处理和保养组(C),对照组(D),同时利用H E染色观察比较A和D组细胞小片显微结构的...     

低分子量氨基糖及其衍生物对角膜细胞生长影响的研究

体外培养兔角膜上皮细胞和基质细胞,采用显微观察和MTT相结合的方法分析低分子量氨基糖(包括壳寡糖(chitosan)、羧甲基壳寡糖(Carboxymethyl chitosan oligosaccharide)、羧甲基甲壳寡糖(Carboxymethyl chitin oligosaccharide)、N-乙酰氨基葡萄糖(N-acetyl-glucosamine))对细胞生长的影响。结果在0~1000μg/mL浓度范围内4种氨基糖对角膜上皮细胞和基质细胞均没有细胞毒性,且均能促进角膜上皮细胞的生长,以壳寡糖和羧甲基甲壳寡糖效果最佳,二者与对照组相比均具有显著性差异(P<0.05)。壳寡糖、羧甲基甲壳寡糖和N-乙酰氨基葡萄糖能明显促进角膜基质细胞的生长,与对照组相比具有显著性差异(P<0.05),其中壳寡糖的促进作用最明显。提示低分子量氨基糖可适用于角膜细胞促生长的培养,为壳聚糖衍生物材料用于眼科研究提供一定的实验依据。     

淫羊藿苷元自微乳在Caco-2细胞模型的肠吸收特性初步研究

目的 制备淫羊藿苷元自微乳,考察其理化性质及体外肠吸收特性.方法 在考察淫羊藿苷元理化性质的基础上,以油酸乙酯为油相,聚山梨酯80为乳化剂,丙三醇为助乳化剂制备淫羊藿苷元自微乳,采用透射电镜及激光粒度分析仪测定其稀释后所得微乳的形态、粒径分布和Zeta电位,并采用Caco-2细胞模型初步分析其肠吸收特性.结果 所制备的淫羊藿苷元自微乳稀释后,微乳平均粒径为55.6 nm,Zeta电位为-30.8 mV,在Caco-2细胞模型上的表观渗透系数(Papp)为(3.52±0.3)×10-6 cm/s.结论 淫羊藿苷元自微乳制剂稳定,体外研究显示自微乳系统能够促进淫羊藿苷元在肠道的吸收.     

Generation of Reactive Oxygen Species in Different Fractions of the Coelomocytes of Holothurian Eupentacta fraudatrix in Response to the Thermostable Toxin of Yersinia pseudotuberculosis in Vitro

Pure fraction (92% - 95%) of phagocytes (FP) and a mixture of amoebocytes (62%) and morula cells (38 % )-FPMC- of the holothurian Eupentacta fraudatrix ( Holothuroidea,Dendrochirota ) were obtained by using ficoll-verographine step gradient. Basal production of reactive oxygen species (ROS) in FP quantified by using reduction of nitroblue tetrazolium (NBT) was more than twice that in FPMC. Thermostable toxin of Yersinia pseudotuberculosis (TST) at differentconcentrations ( 0.2; 0.5; 2.5 μg/ml, but not 0.1 μg/ml) stimulated NBT reduction in FPMC after 24 h incubation. In FP, TST at concentrations of 0.1 and 0.2 μg/ml inhibited and at concentrations of 0.5 and 2.5 μg/ml stimulated NBT reduction after 24 h incubation. Maximal effect was observed in FP and FPMC at TST concentrations of 0.5 and 0.2 μg/ml, respectively. Addition of catalase (0.7 μg/ml) to the cells treated with TST (2.5 μg/ml) was followed by a decrease in NBT reduction compared to that under toxin treatment alone. TST stimulated superoxide dismutase activity in ccncentration-dependent manner (maximum at 0.5 μg/ml concentration in FP) after 24 htreatment, and this stimulation was prevented by a commercial catalase. Plant lectin concanavalin A stimulated NBT reduction more than 5-fold in FPMC compared to the control. With addition of TST, lectin stimulated ROS to lesser extent than that with lectin alone. When catalase, TST, and lectin were added into the FPMC simultaneously, ROS increase was similar to that under lectin treatment alone. On the whole, data obtained indicated that ROS generation in holothurian coelomocytes especially occurs in both stimulated and not stimulated phagocytes, and that changes in ROS production by these cells may be one of the mechanisms of antibacterial protection of holothurians.