Immune condition of Apostichopus japonicus during aestivation

Aestivation is an indispensable state in the life history of sea cucumbers, Apostichopus japonicus. The immune characteristics of the coelomic fluid of A. japonicus, were investigated during aestivation. Samples were collected between July and November 2006 from a coastal pond located off the Yellow Sea in Jiaonan, Shandong Province, China. The total coelomocytes counts (TCC), total superoxide dismutase (T-SOD), catalase (CAT), myeloperoxidase (MPO), and lysozyme (IZM) in the coelomic fluid were measured. The activities of catecholamines, [adrenaline (AD), noradrenaline (NA), and dopamine (DOP)] were estimated. TCC decreased from July to September, indicating weakness of the cellular immune activity at that time. Activities of SOD, CAT, MPO, and LZM changed significantly from July to October. Catecholamines AD and NA in coelomic fluid were significantly higher on August 21 and November 27. There was no significant variation in DOP during the sampling period. Thus, immune characters in coelomic fluid of A. japonicus changed significantly during aestivation. Water temperature was significantly and negatively correlated with TCC, and salinity was significantly and positively correlated with AD and NA. The mechanism of aestivation in A. japonicus is complex and might not be attributed only to environmental changes, such as temperature and salinity, as shown in previous studies. (C) 2008 Elsevier B.V. All rights reserved.

Formation and structure of composite coating of HDA and micro-plasma oxidation on A3 steel

Composite coatings were obtained on A3 steel by hot dipping aluminum(HDA) at 720 ℃ for 6 min and micro-plasma oxidation(MPO) in alkali electrolyte. The surface morphology, element distribution and interface structure of composite coatings were studied by means of XRD, SEM and EDS. The results show that the composite coatings obtained through HDA/MPO on A3 steel consist of four layers. From the surface to the substrate, the layer is loose Al2O3 ceramic, compact Al2O3 ceramic, Al and FeAl intermetallic compound layer in turn. The adhesions among all the layers are strengthened because the ceramic layer formed at the Al surface originally, FeAl intermetallic compound layer and substrate are combined in metallurgical form through mutual diffusion during HDA process.Initial experiment results disclose that the anti-corrosion performance and wear resistance of composite coating are obviously improved through HDA/MPO treatment.

Formation and Structure of Composite Coating of HAD and Micro-Plasma Oxidation on A3 Steel

Composite coatings were obtained on A3 steel by hot dipping aluminum(HAD) at 720 degreesC for 6 min and micro-plasma oxidation (MPO) in alkali electrolyte. The surface morphology, element distribution and interface structure of composite coatings were studied by means of XRD, SEM and EDS. The results show that the composite coatings obtained through HAD/MPO on A3 steel consist of four layers. From the surface to the substrate, the layer is loose Al2O3 ceramic, compact Al2O3 ceramic, At and FeAl intermetallic compound layer in turn. The adhesions among all the layers are strengthened because the ceramic layer formed at the At surface originally, FeAl intermetallic compound layer and substrate are combined in metallurgical form through mutual diffusion during HAD process. Initial experiment results disclose that the anti-corrosion performance and wear resistance of composite coating are obviously improved through HAD/MPO treatment.

栉孔扇贝免疫性的研究

自1997年3月至1999年2月，从青岛麦岛海珍品养殖场购买栉孔扇贝，利用普通组织学、组织化学、透射电镜、扫描电镜、电镜细胞化学和生物化学等技术，对栉孔扇贝血细胞的形态、血淋巴中参与免疫防御的酶的活力和免疫刺激剂对血淋巴中酶活力的影响进行了研究。将其血细胞分为：拟淋巴细胞、血栓细胞、中性粒细胞、嗜碱性粒细胞、嗜酸性粒细胞和巨噬细胞6类。中性粒细胞对细菌的吞噬能力最强，嗜碱性粒细胞吞噬能力较弱，嗜酸性粒细胞无吞噬能力。无颗粒细胞中只有少量较大的细胞具有吞噬能力。扫描电镜观察栉孔扇贝血细胞有圆形、椭圆形和梭形3种形状。多数血细胞容易变形形成伪足。可形成大量长的纤维状伪足的血细胞具有凝血功能。血细胞溶酶体呈强的酸性磷酸酶（ACP）阳性。碱性磷酸酶（AKP）阳性颗粒常沿细胞膜分布。生化测定，血细胞和血清中均存在ACP和AKP活性。血清中ACP和AKP的最适pH值分别为4.4和10.3，最适温度分别为35 ℃和55 ℃。血细胞质中直径分别为300～430nm 和130～200nm的大、小两种颗粒呈强的酚氧化酶（PO）阳性；次级溶酶体和小颗粒呈旨的髓过氧物酶（MPO）阳性。生化测定，血清中存在PO和MPO活性，血细胞仅有MPO活性。血清中PO和MPO的最适pH值分别为6.0和6.5，最适温度分别为45 ℃和35 ℃。血细胞中超氧化物歧化酶（SOD）和过氧化氢酶（CAT）活力分别为312.7U和124.4U，血清中SOD和CAT活力分别为102.3U和73.8U；血细胞中CuZn-SOD和Mn－SOD的活力分别为其SOD总活力的70.3％和29.7％，血清中仅存在CuZn－SOD活性。脂多糖、硒多糖和酵母聚糖A对栉孔扇贝血清和血细胞中溶菌酶、ACP、AKP、PO、MPO、SOD和CAT7种酶的活力均有激活加强作用。

皱纹盘鲍血细胞内活性氧抗病机制及几种环境污染物及其影响的研究

本研究首次揭示了皱纹盘鲍(Haliotis discus hannai) 血细胞内存在着利用活性氧（Reactive oxygen species, ROS）的抗病机制。为今后我国研究贝类的活性氧抗病机制和筛选免疫药物提供了理论基础。本研究主要结果如下：1. 利用鲁米诺（Luminol, 3-氨基邻苯二甲酰肼）为依赖的化学发光法在体外条件下用不同刺激物（酵母细胞和酵母聚糖）对皱纹盘鲍血细胞进行刺激，测定血细胞吞噬活动中历经呼吸爆发产生活性氧的化学发光反应。结果表明皱纹盘鲍血细胞在体外条件下，经刺激物诱导吞噬活动中有明显的呼吸爆现象和很强的活性氧产生。不同有刺激物诱导血细胞产生的化学发光强度不同；同一刺激物的不同处理和不同浓度对血细胞产生活性氧的化学发光强度的影响不同。刺激物经皱纹盘鲍自体血清调理和未经调理对血细胞刺激所产生的化学发光强度不同。SOD和NaN_3对皱纹盘鲍血细胞吞噬过程中活性氧产生的化学发光有抑制作用。上述结果证明了皱纹盘鲍血细胞内存在有象高等动物血细胞内一样的MPO-H_2O-卤素系统的氧化性抗病机制，即在血细胞吞噬异物过程中能够释放活性氧（ROS）对异物进行杀灭的功能。2. 利用活性氧清除剂（过氧化氢酶、SOD、苯甲酸钠、2，5-二甲基呋喃、NBT和EDTA等）对皱纹盘鲍血细胞释放的活性氧进行分类测试，结果表明活性氧清除剂对皱纹盘鲍血细胞吞噬的化学发光都有明显的抑制作用，从而证明皱纹盘鲍血细胞能够释放的活性氧的种类有：超氧阴离子（O_2~-），过氧化氢（H_2O_2），羟自由基（OH~·）和单线态氧（~1O_2）。3. 在体外条件下利用化学发光法定量地研究了不同种类和不同浓度的农药：对硫磷（Parathion）、敌敌畏（Dichlorovos)、乐果（Dimethoate)、2，4-D 丁酯(2,4-D butylester)和甲胺磷（Methamidophos）对皱纹盘鲍血细胞氧化性抗病机制的影响。结果显示不同农药对皱纹鲍血细胞产生ROS的影响程度不同，及同一种农药的不同浓度的浸泡1h或浸泡12h处理皱纹盘鲍血细胞都能够抑制血细胞吞噬时的化学发光，表明农药能够抑制皱纹敌国鲍血细胞吞噬活动中的活性氧（ROS）的产生，而且这种抑制作用随着农药浓度的增加而加强。几种农药抑制皱纹盘鲍血细胞产生活性氧（ROS）的强度不同：2，4D-丁酯，敌敌畏和乐果的抑制作用强于对硫磷和甲胺磷。同时测定了不同浓度的农药浸泡12h后的皱纹盘鲍因细胞吞噬酵母细胞的吞噬百分率和吞噬指数，结果显示多数农药在低浓度时能够提高皱纹盘鲍血细胞的吞噬百分率和吞噬指数，而在高浓度时则能抑制血细胞吞噬酵母细胞的活力而降低血细胞的吞噬百分率的吞噬指数。4. 在体外条件下利用化学发光法定量地研究了不同种类和不同浓度的重金属：铭（Cr）、镉（Cd）、汞（Hg）、铅（Pb）、铜（Cu）和锌（Zn）对皱纹盘鲍血细胞氧化性抗病机制的影响。结果显示不同种类和不同浓度的重金属浸泡1hr.处理皱纹盘鲍血细胞都不同程度地抑制了血细胞吞噬时的化学发光，表明重金属能够抑制皱纹盘鲍血细胞吞噬活动中的活性氧（ROS）的产生，而且这种抑制作用随着重金属浓度的增加而加强。不同的重金属抑制强度不同，从强到弱依次为Hg > Cd > Cu > Pb > Cr > Zn. 研究表明六种重金属中，Hg对皱纹盘鲍血细胞的免疫毒性最大。

Growth, survival and immune activity of scallops, Chlamys farreri Jones et Preston, compared between suspended and bottom culture in Haizhou Bay, China

We examined the growth, survival and immune response of the scallop, Chlamys farreri, during a 1-year period in deep water of Haizhou Bay. Scallops were cultured using two methods: (1) in lantern nets at a 5 m depth and (2) in a bottom culture system (sleeves) on the seabed at about a 25 m depth. Shell heights, meat dry weight and immune activities in the haemolymph (superoxide dismutase and myeloperoxidase) were measured bimonthly or quarterly from July 2007 to June 2008. Survival was measured at the end of the study and environmental parameters in the experimental layers were monitored during the experiment. The growth and immune activities of scallops were lower when the water temperature was high, which was consistent with the main mortality occurring in summer. The growth and immunity of scallops were higher in the suspended culture than in the bottom culture during the experiment, with the exception of shell growth during the last study period. Survival of scallops in the suspended culture (54.6 +/- 12.3%) was significantly lower than that in the bottom culture (86.8 +/- 3.5%) at the end of this study. We conclude from our results that the high mortality of C. farreri can be prevented by culturing them in a bottom culture system before November of the first year, and then transferring them to a suspended culture to improve scallop production.

Effects of acute temperature or salinity stress on the immune response in sea cucumber, Apostichopus japonicus

Invertebrates are increasingly raised in mariculture, where it is important to monitor immune function and to minimize stresses that could suppress immunity. The activities of phagocytosis, superoxide dismutase (SOD), catalase (CAT), myeloperoxiclase (MPO), and lysozyme (LSZ) were measured to evaluate the immune capacities of the sea cucumber, Apostichopus japonicus, to acute temperature changes (from 12 degrees C to 0 degrees C, 8 degrees C, 16 degrees C, 24 degrees C, and 32 degrees C for 72 h) and salinity changes (from 30 parts per thousand to 20 parts per thousand, 25 parts per thousand, and 35 parts per thousand for 72 h) in the laboratory. Phagocytosis was significantly affected by temperature increases in 3 h, and by salinity (25 parts per thousand and 35 parts per thousand) changes in 1 h. SOD activities decreased significantly in 0.5 h to 6 h samples at 24 degrees C. At 32 degrees C, SOD activities decreased significantly in 0.5 h and 1 h exposures, and obviously increased for 12 h exposure. CAT activities decreased significantly at 24 degrees C for 0.5 h exposure, and increased significantly at 32 degrees C in 3 h to 12 h exposures. Activities of MPO increased significantly at 0 degrees C in 0.5 h to 6 In exposures and at 8 degrees C for 1 h. By contrast, activities of MPO decreased significantly in 24 degrees C and 32 degrees C treatments. In elevated-temperature treatments, activities of LSZ increased significantly except at 32 degrees C for 6 h to 12 h exposures. SOD activity was significantly affected by salinity change. CAT activity decreased significantly after only 1 h exposure to salinity of 20 parts per thousand.. Activities of MPO and LSZ showed that A. japonicus tolerates limited salinity stress. High-temperature stress had a much greater effect on the immune capacities of A. japonicus than did low-temperature and salinity stresses. Crown Copyright (C) 2008 Published by Elsevier Inc. All rights reserved.

中低纬电离层暴的统计分析与经验建模

As a key issue of ionospheric weather study, systemic studies on ionospheric storms can not only further improve our understanding of the response of the ionosphere to solar and geomagnetic disturbances, but also help us to reveal the chemical, dynamic and electro-dynamic mechanisms during storms. Empirical modelling for regional ionospheric storm is also very useful, because it can provide us with tools and references for the forecasting and further practical application of ionospheric activity. In this thesis, we focus on describing and forecasting of ionospheric storms at middle and low latitudes. The main points of my investigations are listed as follows. (1) By using magnetic storms during the period over 50 years, the dependence of the type, onset time and time delay of the ionospheric storms on magnetic latitude, season and local time at middle and low latitudes in the East-Asian sector are studied. The results show that the occurrences of the types of ionospheric disturbances differ in latitude and season. The onset of the ionospheric storms depends on local time. At middle latitudes, most negative phase onsets are within the local time interval from night to early morning, and they rarely occurred in the local noon and afternoon sectors. At low latitudes, positive phases commence most frequently in the daytime sector as well as pre-midnight sector. The average time delays for both the positive and negative ionospheric storms increase with descending latitudes. The time delay has significant dependence on the local time of main phase onset (MPO). The time delay of positive response is shorter for daytime MPO and longer for night-time MPO, whereas the opposite applies for negative response. (2) Based on some previous researches, a primary empirical model for mid-latitude ionospheric disturbance is set up. By fitting to the observed data, we get a high accuracy with a mean RMSE of only 12-14% in summer and equinox. The model output has been compared with the output of STORM model, and the results show that, our model is much better than STORM in summer and a little better for some mid-latitude stations at equinox. Especially, for the type of two-step geomagnetic storm, our model can present twice descending of foF2 very well. In addition, our model can forecast positive ionospheric storms.