Adenovirus Carrying Gene Encoding Haliotis discus discus Sialic Acid Binding Lectin Induces Cancer Cell Apoptosis

Lectins exist widely in marine bioresources such as bacteria, algae, invertebrate animals and fishes. Some purified marine lectins have been found to elicit cytotoxicity to cancer cells. However, there are few reports describing the cytotoxic effect of marine lectins on cancer cells through virus-mediated gene delivery. We show here that a replication-deficient adenovirus-carrying gene encoding Haliotis discus discus sialic acid binding lectin (Ad.FLAG-HddSBL) suppressed cancer cell proliferation by inducing apoptosis, as compared to the control virus Ad.FLAG. A down-regulated level of anti-apoptosis factor Bcl-2 was suggested to be responsible for the apoptosis induced by Ad.FLAG-HddSBL infection. Further subcellular localization studies revealed that HddSBL distributed in cell membrane, ER, and the nucleus, but not in mitochondria and Golgi apparatus. In contrast, a previously reported mannose-binding lectin Pinellia pedatisecta agglutinin entered the nucleus as well, but did not distribute in inner membrane systems, suggesting differed intracellular sialylation and mannosylation, which may provide different targets for lectin binding. Further cancer-specific controlling of HddSBL expression and animal studies may help to provide insights into a novel way of anti-cancer marine lectin gene therapy. Lectins may provide a reservoir of anti-cancer genes.     

环境因子对鲍和牡蛎精子运动能力及受精率的影响

研究了环境因子如K^ 、Na^ 、Ca^2 、Mg^2 、Cl^-、SO4^2-以及蔗糖、pH、氨海水对皱纹盘鲍（Haliotis discus Hannai）和太平洋牡蛎（Creastrea gigas）精子运动及受精率的影响。精子在0.4-0.5mol/L的NaCl、KCl溶液中运动能力为1005，但受精能力较差或完全不受精。在替代海水中无Ca^2 的情况下，牡蛎不受精，但鲍有较低的受精能力。Mg^2 作用不明显。在0.5-0.6mol/L蔗糖溶液中鲍精子能运动，且有较低的受精能力，但牡蛎精子完全不能运动和受精。氨海水对精子运动有一定的刺激作用。在pH为6.0-8.0时精子运动能力较强，但受精的pH适宜范围为7.0-8.0，同时鲍和牡蛎在精子运动和受精方面有明显差异。     

饵料对不同规格皱纹盘鲍能量收支的影响

对3种不同规格的皱纹盘鲍(41.40±2.05、54.22±2.66、63.17±2.52mm)分别进行不同饵料搭配的投喂,并对各生理活动的能量代谢进行了测量与计算。实验结果表明,搭配投喂的实验组中皱纹盘鲍能够摄取更多的有机物作为能量代谢的基质。扣除代谢能、排泄能和排粪能后,搭配投喂能提供给皱纹盘鲍的生长能显著高于对照组(P<0.05),尤其是孔石莼与裙带菜搭配投喂组以及裙带菜与海带搭配投喂组,其获得的生长能的比例在3种规格组中均处在很高的水平,是鲍的筏式养殖中值得推广使用的投喂方法。     

Antioxidant and Angiotensin I Converting Enzyme Inhibition Effects and Antihypertensive Effect in Spontaneously Hyertensive Rats of Peptide Isolated from Boiled Abalone By-Products,Hallotis discus hannai

ABSTRACT

Abalone is a marine gastropod that is an important fishery and food industrial resource, massively maricultured in Asia, Africa, Australia, and America. Marine by-products from boiled abalone, normally discarded as industrial waste in manufacturing plants, were fractionated and concentrated using an ultrafiltration membrane system (? 1 kDa). In this study, the antioxidant and angiotensin I converting enzyme (ACE) inhibition effects of the boiled abalone by-products (BABs) were investigated. BABs have high antioxidant activities against free radicals (1,1-diphenyl-2-pycryl-hydrazyl (DPPH), hydroxyl, peroxyl, and superoxide radicals), reactive oxygen species (ROS) stress, and DNA damage in H₂O₂-treated RAW 264.7 macrophages. The structure of the purified peptide was identified to be ATPGDEG (MW 752 Da), and the ACE inhibition pattern of the peptide was found to be noncompetitive. In addition, the peptide exhibited an antihypertensive effect according to time-course measurement after oral administration to spontaneously hypertensive rats (SHR). BABs from this genus may be potential candidates for the development of unique natural substances for further industrial applications as functional foods and pharmaceuticals.     

底栖硅藻种类组成对皱纹盘鲍匍匐幼体和稚鲍生长、存活的影响

为验证由生产培养获得的不同质量的底栖硅藻培养皱纹盘鲍不同生长阶段幼体的效果,采用经不同培养措施获得的两类底栖硅藻群落(饵料A和饵料B)培养皱纹盘鲍前期匍訇幼体和稚鲍.试验结果表明,饵料A培养前期匍匐幼体的存活率[(43.47±13.53)%]显著高于饵料B[(6.94±5.17)%],但生长指标差异不显著.两类饵料培养的稚鲍存活率差异不显著,但饵料B的生长速度和特定生长率[(0.151±0.025)mm/d和(4.706±0.492)%/d)]显著高于饵料A[(0.103±0.022)mm/d和(3.625±0.374)%/d)].说明皱纹盘鲍幼体在不同生长阶段适应底栖硅藻的种类不同.匍匐幼体期对底栖硅藻质量要求较高,稚鲍期由于食量增加和食性转化,对饵料数量的要求表现得更为明显.     

皱纹盘鲍肌动蛋白基因启动子的克隆和序列分析

从皱纹盘鲍雌性个体的足部肌肉提取总DNA后，通过聚合酶连式反应（PCR）技术扩增得到一个扩增产物。经克隆、筛选、确定重组子产物。测序得到了长度为511bp的启动子片段。分析测序结果发现，皱纹盘鲍肌动蛋白基因启动子DNA序列与目前己知的红鲍相应序列的相似度为95%；GC碱基含量为38.93%，较红鲍的低（59.2%）；所得序列含有高度保守的基本表达调控元件，即一个CAAT框和四个TATA框。     

不同卵色皱纹盘鲍(Haliotis discus hannai)幼鲍的生长与免疫比较

随机选取100只经过480 d养殖的3♀×10♂绿卵全同胞家系子一代(F1)的皱纹盘鲍(Haliotis discus hannai)幼鲍为实验材料,进行雌雄、卵色分类后,测量所有样本总重、壳长、壳宽等生长指标,并测定不同卵色幼鲍的超氧化物歧化酶(SOD)、碱性磷酸酶(AKP)、酸性磷酸酶(ACP)、过氧化氢酶(CAT)和溶菌酶(LZM)的活性,进行单因素方差分析(One-way ANOVA).结果显示,在生长方面,不同卵色幼鲍的各生长指标无显著差异(P＞0.05),但绿卵幼鲍的各生长指标均高于其他3种卵色;在免疫酶活性方面,4种卵色幼鲍在SOD和CAT活性上无显著差异(P＞0.05),绿卵幼鲍的AKP、ACP和LZM 3种酶的活性与棕色和棕红色卵幼鲍存在显著差异(P＜0.05),而与棕绿色卵幼鲍无显著差异,其酶活性从高到低按卵色排列依次为绿色、棕绿色、棕色和棕红色.上述研究结果可为利用卵色选育皱纹盘鲍高产抗逆新品系提供依据.     

总氨、亚硝酸氮及养鲍污水对皱纹盘鲍生长的影响

单因子总氨氮(TNH3-N)、亚硝酸氮(N02^--N)试验表明,当养鲍用水中非离子氨氮(NH3-N)为0．035mg／L以下或N02^--N为0．8mg／L以下时,对皱纹盘鲍Haliotis discus hannai Ino幼鲍的生长不产生明显影响;NH3-N对幼鲍生长产生影响的EC5(引起幼鲍生长速度减慢5％的浓度)为0．103mg／L,不产生影响的最大浓度为0．045mg／L。双因素试验结果表明,水中TNH3-N、N02^--N对幼鲍生长的影响没有交互作用,只是简单的相加作用。经回归表明,养鲍自污染水中,NH3-N是影响幼鲍生长的主要因子,它对幼鲍生长产生影响的EC5为0．022mg／L,不产生影响的最大浓度为0．0lmg／L,均比单因子试验结果显著降低。     

皱纹盘鲍维生素D营养需要的研究

在以酪蛋白和明胶为蛋白源的精制饲料中添加不同浓度梯度的维生素D,并以海带为对照组饲喂皱纹盘鲍幼鲍103d。结果表明:幼鲍的生长及软体部水分,脂肪和蛋白质含量受到饲料中维生素D添加水平的影响显著,但成活率不受饲料中添加水平的影响;饲料中适量添加维生素D可以提高幼鲍软体部碱性磷酸酶的活力,但饲料中缺乏或过高水平的维生素D导致幼鲍软体部碱性磷酸酶活力的降低;贝壳中灰分及钙、磷含量受饲料中维生素D添加水平的影响显著。以增重,蛋白质增量和软体部碱性磷酸酶的活力为指标,幼鲍饲料中维生素D的适宜含量为100IU·(100g)-1饲料。