霞水母糖蛋白抗疲劳作用的实验研究

本文对霞水母糖蛋白的抗疲劳作用进行了研究。试验中分别以50、100、200mg／kg剂量的霞水母糖蛋白经口给予小鼠连续灌胃30d,然后分别进行小鼠负重游泳试验、血清尿素氮、肝糖原及血乳酸测定。结果显示,霞水母糖蛋白各剂量组小鼠的游泳时间明显长于对照组（P〈0．01）;各剂量组小鼠运动后血清尿素氮含量低于对照组（P〈0．05或P〈0．01）;中、高剂量组小鼠肝糖原含量均明显高于对照组（P〈0．01）;各剂量组小鼠运动后的血乳酸含量均小于对照组（P〈0．01）。从而表明,霞水母糖蛋白具有抗疲劳作用。     

福建东山岛海域霞水母的渔业生物学研究

1994年4～9月调查结果表明,东山湾海域霞水母的密度高峰期出现在6月25日～7月10日,生物量高峰期在7月10～8月5日;群体的伞径分布范围15～580mm,平均247．48mm,体重分布范围0．4～10250g,平均1274g;高次方程模拟的伞径生长方程式为Φt=15．1726+16．7204t+0．1926t2-0．0014t3—0．0004t4,指数高次方程模拟体重生长方程式为logeWt=-0．6885+1．3397t-0．0959t2+0．0034t3—0．00005t4;以生物经济学原理确定合理开捕为7月15日,合理开捕伞径为360mm．还讨论了表层水温和盐度与霞水母生物量和密度的关系及其资源开发等问题．     

霞水母胶原蛋白活性肽和寡肽胶原抗疲劳作用的实验研究

本文对霞水母胶原蛋白活性肽和寡肽胶原的抗疲劳作用进行了研究。试验中分别以25、50、100mg/kg剂量的霞水母胶原蛋白活性肽和寡肽胶原给小鼠连续灌胃30d,然后进行小鼠负重游泳试验、血清尿素氮、肝糖原及血乳酸含量测定。结果显示,两种受试物各剂量组小鼠的游泳时间明显长于对照组(P0.05或P0.01),各剂量组小鼠运动后血清尿素氮含量及乳酸曲线下面积均低于对照组(P0.01);各剂量组的小鼠游泳后肝糖原含量和空白对照组比较均有不同程度的提高。从而表明,霞水母胶原蛋白活性肽和寡肽胶原均具有抗疲劳作用。     

白色霞水母生活史的实验室观察

本文首次描述了白色霞水母从受精卵至碟状体的生活史。(1)包括受精卵、卵裂、囊胚以及浮浪幼虫等在内的胚胎发育各期均在开放的水体中,在20·8 -21·4℃浮浪幼虫于受精后14 h出现; (2)浮浪幼虫在定置前形成一种凸面的圆形浮浪幼体囊,除了浮浪体囊外,螅状体还可产生足囊和通过产生匍匐茎形成囊胞进而发育成新的螅状体; (3)尽管偶而产生2个碟状体但仍为典型的单碟型横裂; (4)新释放的碟状幼体绝大多数为8个缘叶, 8个感觉棍和8对钝圆的缘瓣,但畸形个体最多12个,最少6个缘叶; (5)雌雄个体间的交互作用对产卵和受精是非常重要的因子[动物学报52 (2) : 389 -395 , 2006]。     

具有ACE抑制活性的霞水母酶解肽制备条件优化

目的:采用胰蛋白酶制备霞水母ACE抑制肽,并以响应面法优化酶解肽制备工艺,通过超滤分离获得具有ACE抑制作用的酶解肽活性部位。方法:单因素实验以ACE抑制率为指标,考察温度、酶解时间、pH、加酶量,进一步通过响应面法优化酶解工艺,以不同截留分子量的纤维膜进行分离。结果:胰蛋白酶水解霞水母制备ACE抑制肽的最佳工艺条件为:酶解温度:49.6℃,加酶量为0.69%,pH为7.8,酶解时间2.0h,酶解产物的平均ACE抑制率为56.79%,与预测值57.20%的相对误差为0.41%,经过超滤分离获得分子量1k Da的酶解产物,ACE抑制活性最高,其IC50为66.58μg/ml。结论:确定霞水母酶解肽的制备工艺,其中分子量1kDa的部分为主要效应物质基础。     

安徽省桃花水母的初步研究

桃花水母属Craspedacusta隶属于刺胞动物门Cnidaria水螅纲Hydrozoa淡水水母目Limnomedusae笠水母科Olindiidae。我国的桃花水母种类丰富,分布广泛,近年来多地有新种和分布新纪录的报道。安徽曾发现桃花水母,但均未进行详细描述和研究。本文对采自安徽省金寨县梅河河谷水潭内的桃花水母进行了初步研究,依据其伞形、缘膜宽度、触手及平衡囊数目、刺丝囊疣形状和排列方式、生殖腺形状和颜色等形态特征对其进行初步的分类鉴定。结果表明,此次在金寨县发现的桃花水母更接近于宜昌桃花水母Craspedacusta kawaii Oka,1907。这是对安徽省发现的桃花水母首次进行较为明确的分类鉴定和初步的形态学研究。     

水母雪莲细胞培养物调血脂作用的初步研究

为研究水母雪莲细胞培养物对高脂大鼠的调血脂作用,将雄性SD大鼠分为4组：正常对照组(A组)、高脂模型组(B组)、高剂量组(C组)和低剂量组(D组)。A组喂基础饲料,B组喂高脂饲料,C组喂高脂饲料的同时饲以大剂量水母雪莲细胞培养物,D组喂高脂饲料的同时饲以小剂量水母雪莲细胞培养物。给药1/d,3周后采血,测定血脂水平及肝肾功能。C组较B组各项血脂指标均有改善,各组间肝肾功能未见显性差异。初步研究表明,水母雪莲细胞培养物对高脂大鼠具有调血脂的作用。本实验用药剂量安全。     

霞水母刺丝囊细胞热激蛋白60(Hsp60)的分离纯化与鉴定

热激蛋白60作为分子伴侣家族中的重要成员,在蛋白质的运输、组装以及折叠等方面起到重要的作用。利用离子交换层析和凝胶过滤层析两步纯化方法,从霞水母刺丝囊细胞中分离到热激蛋白60。SDS-PAGE结果显示,在分子量为60kDa处显示为单一清晰的蛋白条带,并且通过N末端测序进行鉴定,其序列为APKEIKFGADAKSLM与热激蛋白60相吻合;此外,还利用ELISA法对其进一步确定,同时对分离过程的热激蛋白60的回收率进行了测定。该方法为进一步研究霞水母热激蛋白60的功能及其应用奠定了基础。     

温度、投饵频次对白色霞水母无性繁殖与螅状体生长的影响

白色霞水母是我国近海主要大型灾害水母种类之一,其暴发性增殖严重破坏了海洋生态系统平衡.在室内控制条件下,研究了温度(7.5、11、14.5、18、21.5和25℃)和投饵频次(1次/2d、1次/8d和1次/16d)对白色霞水母无性繁殖与螅状体生长的影响.结果显示,白色霞水母足囊繁殖的适宜温度为18-25℃,足囊繁殖随温度和投饵频次的增加而增加.温度对白色霞水母横裂率和横裂次数的影响显著,温度越高,白色霞水母发生横裂生殖的时间越早,横裂生殖速度越快,重复横裂次数越多,释放的碟状体数量也越多.横裂率和横裂次数随投饵频次的增加而递增.白色霞水母螅状体在7.5-25℃范围的成活率均为100％,其生长速度随温度和投饵频次的增加而增加.温度和投饵频次对白色霞水母螅状体足囊繁殖、横裂率和螅状体生长具有明显的交互效应.螅状体的横裂次数和初生碟状幼体伞径随螅状体柄径增大而递增,呈线性相关.研究表明,温度、投饵频次即营养条件显著影响着白色霞水母的种群数量,说明海水水温上升、富营养化或渔业资源锐减导致的浮游动物量增加均可能诱发白色霞水母暴发性增殖.结论为进一步探索大型水母暴发的生态环境机理提供重要科学依据.     

台湾海峡及其邻近海区珍妮水母属的研究(丝螅水母目,面具水母科)

记述了台湾海峡及其邻近海区面具水母科Pandeidae Haeckel,1870珍妮水母属faniopsis Bouillon,1980 5新种,即顶斑珍妮水母faniopsis apicispottis Xu,Huang et Lin,Sp.nov.、短距珍妮水母f.brevispura Xu,Huang et Guo,sp.nov.、大球珍妮水母f.macrobulbosa Xu,Huang et Guo,sp.nov.、南沙珍妮水母f.nanshaensis Xu,Huang et Lin,sp.nov.和蹄形珍妮水母f.unguliformis Xu,Huang et Lin,sp.nov.;并讨论了它们与近缘种的区别,同时将珍妮水母属已知种作个检索表.模式标本保存在厦门大学海洋学系.     

Transglycosylation and reverse hydrolysis reactions of endoglycoceramidase from the jellyfish, Cyanea nozakii

Endoglycoceramidase (EGCase: EC 3.2.1.123) is an enzyme capable of cleaving the glycosidic linkage between oligosaccharides and ceramides in various glycosphingolipids. We report here transglycosylation and reverse hydrolysis reactions of EGCase from the jellyfish Cynaea nozakii. Various alkyl-GM1 oligosaccharides (alkyl-II(3)NeuAcGgOse4) were synthesized when GM1 ganglioside was treated with the EGCase in the presence of 1-alkanols. Among various 1-alkanols tested, methanol was found to be the most preferential acceptor, followed by 1-hexanol and 1-pentanol. GM1 was the best donor, followed by GD1b and GT1b, when methanol was used as an acceptor. However, neither globoside nor glucosylceramide was utilized by the enzyme as a donor substrate. The enzyme transferred oligosaccharides from various glycosphingolipids to NBD-ceramide, a fluorescent ceramide, producing NBD-labeled glycosphingolipids. In addition to the transglycosylation reaction, the enzyme catalyzed the reverse hydrolysis reaction; lactose was condensed to ceramide to generate lactosylceramide in the presence of the enzyme. These results indicate that the jellyfish enzyme will facilitate the synthesis of various neoglycoconjugates and glycosphingolipids.     

Cardiovascular Effect Is Independent of Hemolytic Toxicity of Tentacle-Only Extract from the Jellyfish Cyanea capillata

Our previous studies have confirmed that the crude tentacle-only extract (cTOE) from the jellyfish Cyanea capillata (Cyaneidae) exhibits hemolytic and cardiovascular toxicities simultaneously. So, it is quite difficult to discern the underlying active component responsible for heart injury caused by cTOE. The inactivation of the hemolytic toxicity from cTOE accompanied with a removal of plenty of precipitates would facilitate the separation of cardiovascular component and the investigation of its cardiovascular injury mechanism. In our research, after the treatment of one-step alkaline denaturation followed by twice dialysis, the protein concentration of the treated tentacle-only extract (tTOE) was about 1/3 of cTOE, and SDS-PAGE showed smaller numbers and lower density of protein bands in tTOE. The hemolytic toxicity of tTOE was completely lost while its cardiovascular toxicity was well retained. The observations of cardiac function, histopathology and ultrastructural pathology all support tTOE with significant cardiovascular toxicity. Blood gas indexes and electrolytes changed far less by tTOE than those by cTOE, though still with significant difference from normal. In summary, the cardiovascular toxicity of cTOE can exist independently of the hemolytic toxicity and tTOE can be employed as a better venom sample for further purification and mechanism research on the jellyfish cardiovascular toxic proteins.     

Two-step purification and in vitro characterization of a hemolysin from the venom of jellyfish Cyanea nozakii Kishinouye

Hemolysin is one of the most hazardous components in the venom of Cyanea nozakii Kishinouye. Here we describe the purification and in vitro characterization of the hemolysin, which we named CnPH. The CnPH was isolated by anion-exchange and size-exclusion chromatography from the nematocyst venom. Two protein bands with molecular masses of 20 kDa, 60 kDa respectively were shown in the reducing SDS-PAGE analysis of the CnPH. And Approximately 5 μg/mL of the CnPH resulted in 50% hemolysis of the erythrocyte suspension. The hemolytic activity of the CnPH was both temperature and pH dependent. Moreover, it was significantly inhibited in the presence of divalent metal cations, including Cu²⁺, Mg²⁺, Mn²⁺, Zn²⁺ and Ca²⁺, but enhanced in the presence of EDTA. However, how CnPH performs its hemolytic activity is not yet clear, therefore the mechanism of the hemolytic activity of the CnPH is under research.     

Purification, characterization, and cDNA cloning of a novel acidic endoglycoceramidase from the jellyfish, Cyanea nozakii

Endoglycoceramidase (EC ) is an enzyme capable of cleaving the glycosidic linkage between oligosaccharides and ceramides in various glycosphingolipids. We report here the purification, characterization, and cDNA cloning of a novel endoglycoceramidase from the jellyfish, Cyanea nozakii. The purified enzyme showed a single protein band estimated to be 51 kDa on SDS-polyacrylamide gel electrophoresis. The enzyme showed a pH optimum of 3.0 and was activated by Triton X-100 and Lubrol PX but not by sodium taurodeoxycholate. This enzyme preferentially hydrolyzed gangliosides, especially GT1b and GQ1b, whereas neutral glycosphingolipids were somewhat resistant to hydrolysis by the enzyme. A full-length cDNA encoding the enzyme was cloned by 5'- and 3'-rapid amplification of cDNA ends using a partial amino acid sequence of the purified enzyme. The open reading frame of 1509 nucleotides encoded a polypeptide of 503 amino acids including a signal sequence of 25 residues and six potential N-glycosylation sites. Interestingly, the Asn-Glu-Pro sequence, which is the putative active site of Rhodococcus endoglycoceramidase, was conserved in the deduced amino acid sequences. This is the first report of the cloning of an endoglycoceramidase from a eukaryote.