人参前胡汤治疗椎动脉型颈椎病痰湿中阻证30例临床观察

根据研究证明,在机体先天免疫系统中溶菌酶作为重要的效应分子之一,通过参与机体中多种免疫反应,主要是通过形成一个水解体系,从而破坏和消除侵害体内的病原体,进而促使机体实现免疫防御。本研究以生活在深海超深渊海沟极端环境下的钩虾Eurythenes gryllus为材料,首次从Eurythenes gryllus中克隆获得i型溶菌酶(i-type lysozyme)基因EgLyz。序列分析表明,基因EgLyz与已知序列相似性较低,并含有一个失稳酶结构域。综合分析结果表明EgLyz可能是一个新颖的溶菌酶蛋白。在此基础上,对基因EgLyz进行了大肠杆菌原核重组表达,成功获取了较高纯度的可溶性表达产物,为后续生物活性分析奠定了实验基础。     

Screen and effect analysis of immunostimulants for sea cucumber, <Emphasis Type="Italic">Apostichopus japonicus</Emphasis>

Immunostimulants may improve disease resistance of aquaculture animals by promoting the nonspecific immunity response of the organisms. Five types of saccharides, including chitosan, yeast polysaccharide, burdock oligosaccharide, seaweed polysaccharide and lentinus edodes polysaccharide, were screened for potential use as immunostimulants by using spectrophotometry. The saccharides were injected into Apostichopus japonicus, a sea cucumber, and the lysozyme and superoxide dismutase (SOD) activities of the coelomic fluid and epidermal slime were monitored in six consecutive days. The results show that the lysozyme activity of the animal’s coelomic fluid was significantly stimulated on day 2, day 4 and day 6 after the injection of the saccharides (P<0.05). The effects of chitosan and yeast polysaccharide were the most notable. The lysozyme activity of the epidermal slime was significantly increased by chitosana, yeast polysaccharide, seaweed polysaccharide, and burdock oligosaccharide on day 1 and day 2 after the injection (P<0.05). The SOD activity of the coelomic fluid was significantly promoted by the saccharides on day 2 and day 4 post-injection (P<0.05), while the SOD activity of the epidermal slime increased on day 2. These findings indicate that chitosan and yeast polysaccharide are the most effective immunostimulants and potential healthy anti-disease feedstuff for A. japonicus. Supported by the National Key Technology Research and Development Program (863 Program, No. 2006BAD09A06) and the Special Fund of Chinese Central Government for Basic Scientific Research Operations in Commonweal Research Institutes (No. 02-2007B03)     

海洋真菌药用活性物质研究进展

海洋真菌由于其资源优势和新颖的次生代谢产物,成为海洋天然活性物质的研究热点。按从抗肿瘤、抗菌、酶抑制剂、拮抗剂等几个方面对海洋真菌产生的活性物质的国内外研究情况进行了综述。     

Chemical and structural defensive external strategies in six sabellid worms (Annelida)

In the marine environment, sessile invertebrates have developed an impressive array of mechanisms to avoid predation, bacterial exploitation, and epibiotic overgrowth. In the present study we investigated several defensive strategies adopted by six sabellids: the hard bottom species Sabella spallanzanii (Gmelin, 1791), Branchiomma luctuosum Grube, 1869, Branchiomma bairdi (McIntosh, 1885), and Sabellastarte spectabilis (Grube, 1878), and the soft bottom species Myxicola infundibulum (Renier, 1804), and Megalomma lanigera (Grube, 1846), which have different morphological characteristics and geographical distribution. We examined and compared some defensive features such as branchial crown toughness, tube structure and strength, amount of released mucus, and antibacterial lysozyme‐activity in the mucus. The investigated species utilize a combination of defence and deterrence strategies that seems to be related to the colonized habitat. Tube strength was, higher in the hard bottom species compared with the soft bottom ones, where the tubes are generally buried and protected within the sediment. Branchial crown appeared stronger and resistant in hard bottom species, except for S. spallanzanii, which is the species showing the strongest tube. Sabella spallanzanii, M. infundibulum and S. spectabilis secreted high amount of mucus with high lysozyme‐like activity. By contrast, B. luctuosum, B. bairdi, and M. lanigera produced low amounts of mucus exerting lower antibacterial activity.     

水稻(Oryza sativa L.)秸秆浸提液对铜绿微囊藻(Microcysti aeruginosa)的抑制作用

用刚收割的水稻(Oryza sativa L.)秸秆浸提液代替水配制培养基培养铜绿微囊藻(Microcystic aeruginosa),追踪测定微囊藻的生物量、叶绿素a含量、光合速率、呼吸速率、膜透性、超氧物歧化酶(SOD)活性、光合膜自发荧光强度等相关生理生化指标的变化,研究水稻秸秆浸液对铜绿微囊藻的抑制作用.结果表明,质量比≥1/100(水稻秸秆/水)的水稻秸秆浸提液对微囊藻的生长有明显的抑制作用,表现如下:藻细胞生物量在实验过程中逐日降低,藻细胞叶绿素a含量和光合速率急剧下降,呼吸速率和超氧物歧化酶(SOD)活性呈先升高后下降的趋势,膜透性迅速升高,细胞光合膜自发荧光强度显著衰减.用不同的有机溶剂对该浸提液进行萃取,浓缩后用滤纸片法在固体培养基上做抑藻实验,乙醚和乙酸乙酯萃取液能明显看到抑藻圈,证实其中含有抑制物质.     

一种复方中草药饲料添加剂在红笛鲷网箱养殖中的应用

将黄芪、当归、甘草和山楂等粉碎后按一定比例配伍制成复方中草药饲料添加剂后,按质量分数0.5%、1.0%、1.5%和2.0%添加到基础饲料中,连续投喂红笛鲷(体重234.95±19.26 g,体长24.76±1.42 cm)56 d,研究复合中草药制剂对红笛鲷生长、成活、免疫和抗病力的影响。结果表明,基础饲料中添加本复方中草药饲料添加剂能够显著促进红笛鲷的生长,增强其血清中的抗菌活力和溶菌酶活力,提高红笛鲷成活率及其对溶藻弧菌的免疫保护率,其中以添加1.5%(质量分数)和连续饲喂28 d效果为最佳(P<0.01)。     

僧帽牡蛎碱性磷酸酶性质的研究

从僧帽牡蛎分离纯化出一种碱性磷酸酶,采用Sephadex G-200凝胶过滤柱层析法测得该酶分子量为1.57×10~5.以对硝基苯磷酸为底物,该酶的最适温度为43℃,最适pH值为10.0.在37℃和pH值为10.0的条件下,酶催化反应的米氏常数(K_m)为9.68×10~(-4)mol/dm~3,活化能(E_a)为45.85kJ/mol,温度系数(Q_(10))为1.80(30～40℃).对该酶的热稳定性研究表明:其在45℃以下较为稳定,50℃以上明显失活,并测定了该酶的热失活速度常数.金属离子对酶活力的效应研究结果表明:Mg~(2+)、Mn~(2+)、Ca~(2+)、Co~(2+)对酶有激活作用,Ag~+、Hg~(2+)、Zn~(2+)表现为抑制作用,Cd~(2+)几乎没有影响.Mg~(2+)是较有效的激活剂,表现为部分非竞争性效应,激活常数(K_a)为2.48×10~(-4)mol/dm~3,有Mg~(2+)存在时该酶活性中心的转换数量是无Mg~(2+)时的4.0倍.几种效应物对酶的抑制机理研究表明:HPO_4~(2-)、HAsO_4~(2-)、Cys为竞争性,其抑制常数分别为1.61、1.18和0.15mmol/dm~3;L-Phe为反竞争性,抑制常数为3.26mmol/dm~3.     

BSA-ISCOMs对欧洲鳗鲡免疫效应及抗细菌性病原的效果研究

为证实免疫刺激复合物(ISCOMs)的非特异免疫增强作用,将牛血清白蛋白(BSA)制备成ISCOMs(BSA-ISCOMs).将欧洲鳗鲡(Anguilla anguilla)随机分为3组,分别用BSA-ISCOMs、BSA和磷酸盐缓冲液(PBS,对照组)腹腔注射免疫.二次免疫完成后第30d采血,测血清部分非特异性免疫指标,剩余鳗鲡分别用创伤弧菌(Vibrio vulnificus)和嗜水气单胞菌(Aeromona hydrophila)攻毒.结果显示,BSA-ISCOMs组的血清溶菌酶活力显著高于BSA组和对照组(p≤0.05);超氧化物歧化酶(SOD)和补体C3活力与对照组无显著差异(p0.05).攻毒结果显示,BSA-ISCOMs组的存活率显著高于BSA组和对照组(p≤0.05),对创伤弧菌和嗜水气单胞菌攻击的相对免疫保护率分别为43.3%和51.7%.表明BSA-ISCOMs能有效增强欧洲鳗鲡对主要病原创伤弧菌和嗜水气单胞菌的抗病能力.     

Isolation and Biochemical Characteristics Analyses of Phenoloxidases (POs) in Three Cultured Mollusk Species

Phenoloxidases(POs)are a group of copper proteins including tyrosinase,catecholase and laccase,which play crucial roles in the innate immune response of mollusks.In this research,POs were studied in cultured mollusk species,including scallop Chla-mys farreri,abalone Haliotis discus hannai and clam Scapharca subcrenata.The POs were isolated from hemocytes using linear-gradient native-PAGE combined with catechol staining.The PO activities and their characters were investigated.The molecular mass of PO in C.farreri was 576 kDa,and it was 228 kDa in H.discus hannai.In S.subcrenata,four POs were detected and their mole-cular masses were 391 kDa,206 kDa,174 kDa and<67 kDa,which were named as 391-PO,206-PO,174-PO and s-PO,respectively.Ki-netic analyses indicated that all of the POs,except for 391-PO had higher affinity to L-DOPA and catechol than to hydroquinone and dopamine.However,all of the POs failed to oxidize tyrosine.The effects of divalent metal ions on POs’activities were assayed,in-cluding Fe²⁺,Mg²⁺,Zn²⁺,Mn²⁺,Cu²⁺and Ca²⁺from FeCl₂,MgSO₄,ZnSO₄,MnCl₂,CuSO₄and CaCl₂.The POs were inhibited by Fe²⁺at all determined concentrations.Additionally,the inhibition assay showed that all of the POs were inhibited by cysteine,ascorbic acid,sodium sulfite,citric acid,ethylenediaminetetraacetic acid disodium(EDTA)and sodium diethyldithiocarbamate(DETC).The inhibition effects of critric acid and EDTA are dose-dependent.H.discus hannai PO and 391-PO were slightly inhibited by sodium azide,and H.discus hannai PO,391-PO and 174-PO were slightly inhibited by thiourea.In conclusion,the POs in the three cultured mollusks are copper-containing laccase-type phenoloxidases with similar biochemical characteristics even though their molecular masses are different.     

哈维氏弧菌对条纹斑竹鲨4种酶活性的影响

以哈维氏弧菌（Vibrio harveyi）对条纹斑竹鲨（Chiloscyllium plagiosum）进行病原接种试验,分别在4、8、12、24、48、72、96h后测定肝脏、脾脏、鳃及血清的酸性磷酸酶（acid phosphatase,ACP）、碱性磷酸酶（alkaline phosphatase,AKP）、超氧化物歧化酶（superoxide dismutase,SOD）和血清中溶菌酶（lysozyme,LSZ）的活力.实验结果表明：对照组条纹斑竹鲨的这4种酶的活力呈现出明显的组织差异性,血清中SOD活力为124.32 U/cm3,显著高于其他3种酶的活力;其他组织中3种酶的活力由高到低依次为SOD、ACP、AKP.在实验组条纹斑竹鲨被感染4～72h期间,血清中SOD活力明显下降,ACP活力持续下降,感染96h后除LSZ外,其他3种酶的活力都呈回升趋势;其他组织中ACP与AKP活力变化均为被感染4h后下降,12h后明显回升,SOD则在被感染4h后活力上升,而后下降,96h后回升.这4种酶的活力的变化主要是应激作用与非特异性免疫防御作用共同作用导致的结果.