STRUCTURE OF THIAMINE BROMIDE SESQUIHYDRATE

3-[(4-Amino-2-methyl-5-pyrimidinyl)-methyl]-5-hydroxyethyl-4-methylthiazolium bromide sesquihydrate, C12H17N4OS+.Br-.1.5H2O, M(r) = 372.28, monoclinic, P2(1)/a, a = 11.676 (2), b = 24.819(7), c = 12.344 (3) angstrom, beta = 113.74(2)-degrees, V= 3274 (1) angstrom3 , Z = 8, D(x) = 1.51 g cm-3 (Mo Kalpha) = 0.71069 angstrom, mu = 26.2 cm-1, F(000) = 1528, T 293 K, R = 0.062 for 2720 observed reflections. Both the independent thiamine molecules A and B in the asymmetric unit adopt the common F conformation. A bromide anion is held by four neighbouring thiamine molecules through C(2)-H...Br hydrogen bonds and Br ... thiazolium-ring electrostatic interactions. Another bromide anion (or a water molecule) bridges the pyrimidine and thiazolium moieties of molecule A (or B) through a hydrogen bond with the amino group and an electrostatic interaction with the thiazolium ring.

香豆素440/荧光素钠/藏红混合染料体系的光谱研究

通过对C440/Fl.Na/Saf.-T三元混合染料体系的荧光光谱研究,了解到这三种染料之间有较充分的能量转移。又通过对C440/Fl.Na和Fl.Na/Saf.-T两个混合染料体系的荧光寿命测量,讨论了分子间能量转移过程和机理,并做了一些定量的分析。

Construction of shuttle, expression vector of human tumor necrosis factor alpha (hTNF-alpha) gene and its expression in a cyanobacterium, Anabaena sp. PCC 7120

The construction of the shuttle, expression vector of human tumor necrosis factor alpha (hTNF-alpha) gene and its expression in a cyanobacterium Anabaena sp. PCC 7120 was reported. The 700-bp hTNF cDNA fragments have been recovered from plasmid pRL-rhTNF, then inserted downstream of the promoter PpsbA in the plasmid pRL439. The resultant intermediary plasmid pRL-TC has further been combined with the shuttle vector pDC-8 to get the shuttle, expression vector pDC-TNF. The expression of the rhTNF gene in Escherichia coil has been analyzed by SDS-PAGE and thin-layer scanning, and the results show that the expressed TNF protein with these two vectors is 16.9 percent (pRL-TC) and 15.0 percent (pDC-TNF) of the total proteins in the cells, respectively, while the expression level of TNF gene in plasmid pRL-rhTNF is only 11.8 percent. Combined with the participation of the conjugal and helper plasmids, pDC-TNF has been introduced into Anabaena sg PCC 7120 by triparental conjugative transfer, and the stable transgenic strains have been obtained. The existence of the introduced plasmid pDC-TNF in recombinant cyanobacterial cells has been demonstrated by the results of the agarose electrophoresis with the extracted plasmid samples and Southern blotting with alpha-(32)p labeled hTNF cDNA probes, while the expression of the hTNF gene in Anabaena sp. PCC 7120 has been confirmed by the results of Western blotting with extracted protein samples and human TNF-alpha monoclonal antibodies. The cytotoxicity assays using the mouse cancer cell line L929 proved the cytotoxicity of the TNF in the crude extracts from the transgenic cyanobacterium Anabaena sp. PCC 7120.

QTL Mapping for Frond Length and Width in Laminaria japonica Aresch (Laminarales, Phaeophyta) Using AFLP and SSR Markers

In Laminaria japonica Aresch breeding practice, two quantitative traits, frond length (FL) and frond width (FW), are the most important phenotypic selection index. In order to increase the breeding efficiency by integrating phenotypic selection and marker-assisted selection, the first set of QTL controlling the two traits were determined in F-2 family using amplified fragment length polymorphism (AFLP) and simple sequence repeat (SSR) markers. Two prominent L. japonicas inbred lines, one with "broad and thin blade" characteristics and another with "long and narrow blade" characteristics, were applied in the hybridization to yield the F-2 mapping population with 92 individuals. A total of 287 AFLP markers and 11 SSR markers were used to construct a L. japonica genetic map. The yielded map was consisted of 28 linkage groups (LG) named LG1 to LG28, spanning 1,811.1 cM with an average interval of 6.7 cM and covering the 82.8% of the estimated genome 2,186.7 cM. While three genome-wide significant QTL were detected on LG1 (two QTL) and LG4 for "FL," explaining in total 42.36% of the phenotypic variance, two QTL were identified on LG3 and LG5 for the trait "FW," accounting for the total of 36.39% of the phenotypic variance. The gene action of these QTL was additive and partially dominant. The yielded linkage map and the detected QTL can provide a tool for further genetic analysis of two traits and be potential for maker-assisted selection in L. japonica breeding.

潮间带纤毛虫原生动物和小型底栖动物生态学研究

微型和小型底栖动物是底栖微/小食物网的重要构成。相对浮游生态系统, 迄今国际间对底栖食物网的认知极为欠缺。这一方面是由于微型生物形态和功能上的复杂性和多样性, 另一方面原因在于研究方法的障碍—主要是微型和小型底栖动物的定量提取和定性分析。本研究首先进行了方法学改良, 并应用新方法对底栖微食物网的重要功能类群—纤毛虫原生动物和小型底栖动物进行了不同生境的周年按月采样, 定性及定量研究的同时, 联系环境因子对微型和小型底栖生物的环境监测进行了探讨。 微型和小型底栖生物的定量研究首先涉及到目标生物在沉积物中的有效提取, 目前硅胶液提取是普遍使用的方法, 其中Ludox液主要应用于小型生物, 它不但价格便宜而且比重合适, 因此在常规生态研究中被广为接受。不过, Ludox易与于海水中的阳离子产生凝结而无法直接用于微型生物; 目前唯一直接应用于微型生物提取的是Percoll硅胶液, 但其昂贵的价格使其在常规生态研究中受到极大限制。本研究以价格低廉的Ludox 硅胶液结合定量蛋白银染色 (QPS) 技术开发了一种新的方法, 即Ludox-QPS法。主要流程为: 样品采集与固定、淘洗/稀释降盐、Ludox密度梯度离心、过滤浓缩和琼脂包埋, QPS染色、永久封片及鉴定计数。添加已知数量的纤毛虫至无生物底泥的重获实验表明, 该密度梯度离心的提取率大于94%; 该方法对自然沉积物中纤毛虫的提取率达97.6%, 对沙质、泥沙质和泥质中海洋线虫的提取率分别达97%、96.9% 和97.8%。对比实验表明, 经QPS制片获得的小型动物的丰度和类群数量与传统方法相当或更高, 尤其当小个体虫体占优势时, 该法显示出较传统方法 (导致数量低估) 更为明显的定量优越性。该方法除用于纤毛虫和小型动物的定量分析外, 还具有较高的分类分辨率, 染色后的纤毛虫原生动物大多类群可鉴定到属, 部分可鉴定到种, 以此可在群落水平上研究其生态作用。 根据新开发的Ludox-QPS技术, 在大沽河潮间带依据盐度梯度选定2个站位 (IIQ和营海) 进行了周年按月采样, 对底栖纤毛虫和小型底栖动物进行了定量研究。纤毛虫原生动物在IIQ和营海的年平均丰度分别为2236 inds./10 cm2 和935 inds./10 cm2 (28 inds./ml 和12 inds./ml), 平均生物量分别为119.1 gC/10 cm2和54.2 gC/10 cm2 (1.5 gC /ml 0.7 gC/ml)。丰度的季节变化趋势为: 春天 > 秋天 > 夏天 > 冬天。垂直分布上, 在营海分布于表层0-0.5 cm 的比例为57.1%, 分布于0.5-2 cm、2-4 cm和4-8 cm比例分别为23.1%、11.4% 和8.5%; 13个月中除12月份外, 4-8 cm均有一定数量的纤毛虫分布; 而在IIQ, 97% 的纤毛虫分布在0-0.5 cm, 分布在0.5-2 cm、2-4 cm和4-8 cm比例分别为2.4%、0.4%和0.2%, 4-8 cm的分布只发生在春季和秋季。纤毛虫的多样性季节变化明显, 春秋季物种丰富, 两个站点每毫升沉积物的平均物种数分别为18和6。Two-Way Crossed ANOSIM 分析表明纤毛虫群落在月份间和站点间的差异极其显著。Pseudochilodonopsis sp., Chilodontopsis sp., Euplotes sp.及Prorodon sp.是表征两个生镜中纤毛虫群落的主要类群。 同时, 发现了14个小型生物类群, 其中线虫在IIQ和营海的丰度优势度分别为97.4% 和78.6%。小型动物在IIQ和营海的年平均丰度分别为4793 inds./10 cm2和8915 inds./10 cm2 (60 inds./ml和111 inds./ml), 其生物量分别为1068.8 gC /10 cm2和1790 gC /10 cm2 (13.4 gC/ml和22.4 gC/ml)。小型底栖动物的丰度在IIQ的季节变化为: 夏季 (7888 inds./10cm2) > 秋季 (5447 inds./10cm2) > 春季 (3731 inds./10cm2) > 冬季 (2780 inds./10cm2); 在营海则完全相反: 冬季 (15579 inds./10cm2) > 春季 (10691 inds./10cm2) > 秋季 (6611 inds./10cm2) > 夏季 (4667 inds./10cm2)。小型底栖动物和纤毛虫的相对重要性存在明显的区域和季节差异。 纤毛虫原生动物、小型动物及环境因子的相关分析表明, 纤毛虫的丰度和多样性与温度和盐度及有机质含量显著相关, 与小型动物没有显著相关性; 群落结构分析表明, 温度、有机质和小型动物的丰度的组合与纤毛虫群落丰度的相关系数为0.345; 盐度、脱镁叶绿素、有机质和小型动物生物量的组合与纤毛虫群落多样性的相关系数为0.403。依据海洋线虫和桡足类的比值 (N/C) 推测, IIQ 可能存在严重的有机污染, 营海则存在明显的季节波动, 8月和9月及2月可能是污染最严重的季节, 这种状况在纤毛虫群落结构的CLUSTER聚类中得到验证。虽然目前尚没有形成有关微型底栖生物-纤毛虫原生动物的污染检测的直接依据, 但本研究说明纤毛虫群落的确对环境污染具有一定的感应度, 而且这种感应和利用小型生物的主要类群估算的污染检测 (N/C) 存在一定程度的关联。 90年代早期有关青岛湾有机污染带的研究表明, 经彻底截污后, 其环境状况向良性发展。进一步了解该湾的健康状况, 2006.5-2007.5月对该湾沙质和泥沙质的小型动物进行周年按月采样。小型动物在泥沙质和砂质沉积物中的年平均丰度分别为4853 ± 1292 inds./10 cm2和1528 ± 569 inds./10 cm2; 年平均总生物量分别为1434.1 ± 897.0 gC /10cm2和720.7 ± 353.8 gC/10cm2。沙质底小型生物的丰度季节波动明显, 6月份和12月份最高, 3月份和9月份最低; 泥沙质季节波动不明显, 6月份最高。两个站点均有48%的小型动物分布在0-0.5 cm 表层, 海洋线虫在表层的分布比例分别为48% (泥沙质) 和34% (砂质)。共检获14个小型动物类群, 其中线虫在泥沙质和砂质沉积物中的年平均丰度分别4619 ± 1255 inds./10cm2和1014 ± 376 inds./10cm2, 其丰度优势度分别为95.2%和66.4%。其它在丰度上占优势的类群, 泥沙质依次为多毛类 (1.5%)、甲壳幼体 (1.5%) 和桡足类 (0.7%); 沙质依次为: 甲壳类幼体 (12.6%)、腹毛类 (8.3%) 和 桡足类 (6.2%)。CLUSTER聚类分析表明, 泥沙质和和砂质中小型生物的丰度组成具有64%的相似性。BIOENV分析表明, 温度、盐度、中值粒径和粘土粉砂含量的组合最能解释不同月份之间和不同站位间的差异, 其相关系数为0.614。依据小型生物的丰度和类群组成, 表明泥沙质底尚存一定的有机污染。

中国对虾育种方法研究与遗传连锁图谱的构建

本研究以中国对虾为材料，以杂交育种和选择育种为目标，进行了系统的中国对虾杂交育种试验、生长性状遗传参数试验及其分子遗传连锁图谱的构建工作。结果表明以不同地理群体杂交作为基础群体，然后采用系统的选择育种方法可以获得较好的选择效果。构建的遗传连锁图谱为中国对虾分子辅助育种提供一定的基础。这些试验结果为中国对虾合理系统的育种工作提供了理论基础和数据支持。其具体结果如下： 1. 试验对中国对虾黄渤海水域乳山湾群体（WYP）和朝鲜半岛南海群体（WKN）的2个群体及其杂交后代不同月龄生长情况和存活率进行了研究，测量体长（TL）、头胸甲长（CL）、头胸甲宽（CW）、第2、3腹节高（HST）、第2、3腹节宽（WST）、体重（BW）和存活率共7个性状，计算各项指标的杂种优势率，并对各性状进行了方差分析和多重比较。其3月龄生长情况和存活率研究结果表明，存活率在乳山湾群体（WYP♀）× 朝鲜半岛南海群体（WKN♂）杂交后代出现杂种劣势外，其他指标都表现出不同程度的杂种优势(4.37%～23.96% )。除了存活率外，杂交后代生长性状均显著高于亲本，乳山湾群体（WYP♀）×朝鲜半岛南海群体（WKN♂）杂交后代高于朝鲜半岛南海群体（WKN♀）× 乳山湾群体（WYP♂）杂交后代，黄渤海水域乳山湾群体高于朝鲜半岛南海群体后代。为确定测量性状与中国对虾体重的相关程度，建立了用体长(X1)，头胸甲宽(X2)，第2、3腹节宽(X3)，头胸甲长(X4)，第2、3腹节高(X5)估计体重的多元回归方程：Y = －2.056 + 0.03X1 + 0.076X2 + 0.078X3 + 0.033X4 + 0.043X5。 2. 中国对虾黄渤海水域乳山湾群体（WYP）和朝鲜半岛南海群体（WKN）2个群体及其杂交后代在4月龄时期的6个生长指标和存活率的杂种优势范围在0.514%到14.95%之间，WYP♀×WKN♂杂交后代在这7个指标中都高于WKN♀×WYP♂杂交后代。5月龄杂交后代也表现出一定程度的杂种优势，其范围在-9.000%～19.090%之间，但头胸甲长、第2、3腹节处高和存活率3个指标出现杂种劣势。不同杂交组合各个阶段生长发育情况和存活率在杂种优势表现出一定的规律。随着月龄的增加，WKN♀×WYP♂杂交后代杂种优势率有所增加，而WYP♀×WKN♂杂交后代的却有所降低。ANOVA分析结果表明，杂交后代在存活率方面与双亲差异不显著。4月龄的分析结果发现杂交后代在WST和BW这2个指标上存在显著差异。LSD多重比较结果显示，WYP♀×WKN♂杂交后代在BW指标上与亲本存在显著差异，在WST指标方面与其他3个组合的后代差异显著。5月龄的数据分析结果发现，杂交后代除体重存在显著差异外，其他各项指标差异均不显著。LSD多重比较结果发现，WKN♀×WYP♂杂交后代体重与其亲本WKN存在显著差异。 3. 对2个野生群体——朝鲜半岛南海岸群体（WKN）和黄渤海群体（WYP）和3个养殖群体——朝鲜半岛群体的养殖一代（FKN），黄海1号（HH1）和即抗98（JK98）进行杂交试验的研究，结果表明JK98 (♀)  WKN (♂)组合在存活率方面最高，其余的依次为WYP (♀) WKN (♂)，WKN (♀) WYP (♂)，FKN (♀)HH1 (♂) 和 WYP (♀) FKN (♂)。而在体重方面FKN(♀)  HH1(♂)组合最高，其余的依次为WKN (♀) WYP (♂)，WYP (♀) WKN (♂)，WYP (♀)FKN (♂) 和 JK98 (♀)WKN (♂)。在所有生长性状方面，JK98 (♀)  WKN (♂)在5个组合中是最低的。方差检测结果表明，TL、CL、HST、LL和BW这5个指标在不同组合间存在差异，而其他指标不存在差异。多重比较结果发现JK98 (♀)WKN (♂)组合的TL与其他组合间差异极显著，HST指标与WKN (♀) WYP (♂)，FKN(♀)  HH1(♂)和 WYP (♀)  WKN (♂)这3个组合差异显著，BW指标与WKN (♀) WYP (♂) 和 FKN(♀)  HH1(♂)差异显著。 4. 通过人工授精的方法建立了中国对虾21个半同胞家系，测量了中国对虾21个半同胞（46个全同胞）家系的TL、CL、CW、HST、WST、第1腹节长（FL���、第6腹节长（LL）。利用MTDFREML软件得到生长性状遗传力在0.15~0.35之间，属于中度遗传力范围。TL的遗传力为0.34±0.071，CL的为0.30±0.070，CW为0.35±0.077，WST为0.33±0.073，HST为0.33±0.073，FL���最低为0.15±0.044，LL的为0.24±0.059。各个性状间表现出高的正相关，其中CW和TL以及HST的遗传相关最大，FL���WST的遗传相关最小。 通过以上杂交育种和选择育种的研究，认为单纯的依靠杂交育种来改善中国对虾的育种工作可能具有一定的局限性。所以在实际的育种过程中，以中国对虾不同群体的杂交后代作为基础群体，并以此为基础进行系统的选择育种应该具有更大的潜力。 5. 本试验利用中国对虾F2群体和AFLP分子标记技术进行了遗传连锁图谱的构建。利用55对AFLP引物组合对F2家系的110个个体进行了研究，结果检测到532个符合作图策略的AFLP标记。利用卡方检验检测分离位点是否符合孟德尔分离定律。对于符合3：1比例的分离位点利用F2自交模型构建性别平均连锁图谱，对于符合1：1比例的分离位点利用拟测交理论分别构建中国对虾的雌性和雄性遗传连锁图谱。雌性、雄性和性别平均遗传图谱分别有28、35和44个连锁群，图谱实际长度分别为1090、1617和1772.1 cM。中国对虾遗传连锁图谱估计基因组长度为2420 cM，符合与人类基因组相比的对虾类基因组长度。中国对虾雄性遗传连锁图谱比雌性遗传连锁图谱长32.6%，这可能说明中国对虾不同性别存在不同的重组率。通过皮尔逊相关系数检测认为AFLP标记在中国对虾图谱上分布均匀。本文利用AFLP标记构建的中国对虾遗传连锁图谱为中国对虾基因组研究和遗传改良提供一定的基础，同时也应该开发微卫星等共显性标记，为遗传连锁图谱的整合提供条件。

胶州湾生态系统数学模型与计算机模拟及优化管理对策

六十年代产生的系统生态学，在生态系统研究中使得数学模型至今都是一个必不可少的有效工具，建立数学模型已成为生态学中研究的热点。文章运用概率统计、时序分析手段及灰色系统理论，对胶州湾生态系统调查的历史资料进行初步分析，取得如下成果。1. 建立了非生物环境因子的八个静态统计模型，并分析了它们之间的相关关系，从定性、定量的角度对胶州湾非生物环境因子相互关系进行了研究，并结合实际情况对数学模型进行了解释。2. 根据历史资料及前人研究成果，把胶州湾生态系统划分为：浮游动物、浮游动物食性鱼类、植食性鱼类、底栖生物、底栖生物食性鱼类、肉食性鱼类六个子系统，并把有机质及浮游植物（初级生产力）作为系统的输入，运用灰关联度方法对子系统与生物及非生物环境因子的关系进行定量分析，排列出影响各子系统生物量变化的非生物环境因素的主次顺序，建立了相应的GM（1，N）模型，并对模型进行了残差检验，模型能较好的反映实际情况。预测了各子系统的发展特性，并结合实际情况提出了各类渔业资源管理的具体指标值，对优化子系统提供了具体参数。3. 根据胶州湾的实际情况，提出了新的数学模型 - 差分摆动模型及其建立方法。模型的意义在于可量化任意几个有关因子的数量关系，并对未能考虑到的因子，甚至一直未能被人们认识到的所有因子对这种生物作用的信息量进行量化，把生态系统中生物与非生物环境因子相互作用通过方程联系起来，并在论文中得到了验证。4. 根据灰色系统分析方法，对胶州湾生态系统进行了动态分析，建立了生态系统的量化动态模型，利用劳斯判断准则及根平面法，分析了系统的发展稳定状态，发现胶州湾生态系统周年内处于不稳定的发展状态。总之，论文运用各种数学手段从系统的角度来分析生态系统中各环境因子之间关系，并用数学模型把这种关系定量地表示出来，利用胶州湾生态系统现有资料，对胶州湾生态系统从系统的角度进行综合分析，为科学管理与合理开发胶州湾生物资源、优化胶州湾生态系统提供了意义的参考意见。

迟缓爱德华氏菌基因组fosmid文库的构建、opp基因簇的克隆及aroA基因缺失突变株的构建

本研究构建了迟缓爱德华氏菌(Edwardsiella tarda)LSE40 基因组fosmid文库，该文库共包含2 500个克隆，插入片段平均大小为33.6kb，文库总容量约84Mb，覆盖E.tarda LSE40基因组（按5Mb计算）超过16倍。随机挑取1 000个fosmid克隆进行双末端测序共得到1 741条高质量的序列，序列平均长度546 bp，全长949 997bp，约为E.tarda基因组的19％。将这些序列提交到KEGG自动注释服务器KAAS对所得序列进行代谢途径分析，得到E.tarda LSE40的KO (KEGG Orthology) 注释。分析结果表明，与代谢途径相关的基因有932条序列，与环境信息处理相关基因283条，与遗传信息处理相关基因220条，与细胞进程和人类疾病相关的基因分别为64条和16条。同时将序列进行BlastX，按照微生物致病性共同主题找到61个毒力相关基因。Fosmid文库的建立和部分基因组序列的生物信息学分析为进一步研究E.tarda LSE40的致病机制、代谢机制和生理生态机制提供了丰富的物质基础。 通过比较基因组的方法，从E.tarda LSE40 fosmid文库克隆到编码寡肽透过酶的opp基因簇，该基因簇全长6 741bp，含有5个ORF，依次编码OppA-B-C-D-F 5个蛋白；位于oppA和oppB的间隔区和oppF之后的非编码区各有一个茎环结构，推测分别为oppA和opp基因簇的转录终止子。以细菌OppA的保守结构域SBP_bac_5构建系统发生树，结果显示E.tarda LSE40与同属细菌E.ictaluri的亲缘关系最近，与肠杆菌科细菌的亲缘关系较近，与革兰氏阳性细菌的亲缘关系较远，表明OppA的SBP_bac_5结构域可作为细菌分类鉴定的依据。 从E.tarda LSE40 fosmid文库克隆aroA基因全序列，该序列全长1 287bp，编码428个氨基酸，与鲶鱼爱德华氏菌(E. ictaluri)氨基酸相似性在94%，与其他肠杆菌科菌如Escherichia coli和Yersinia enterocolitica相似性在73%-74%。通过In-frame deletion构建了E.tarda LSE40 aroA缺失突变株。与野生型相比，aroA突变株的半数致死量LD50提高了62倍。在牙鲆接种~106cfu/ml的E.tarda细菌时，接种野生型细菌的牙鲆在6天内全部死亡，濒死鱼的细菌数达7.97×108cfu/ 100mg；而接种aroA突变株的牙鲆没有出现死亡，28天后检测不到细菌的存在。实验结果为进一步评价aroA突变株作为减毒活疫苗打下了基础。

Effects of different light sources and illumination methods on growth and body color of shrimp Litopenaeus vannamei

Shrimps Litopenaeus vannamei with initial body weight of 2.108 +/- 0.036 g were sampled for specific growth rates (SGR) and body color measurements for 50 days under different light sources (incandescent lamp, IL; cool-white fluorescent lamp, FL; metal halide lamp, MHL; and control without lamp) and different illumination methods (illumination only in day, IOD, and illumination day and night, IDN). Body color of L. vannamei was measured according to the free astaxanthin concentration (FAC) of shrimp. The SGR, food intake (FI), feed conversion efficiency (FCE) and FAC of shrimps showed significant differences among the experimental treatment groups (P < 0.05). Maximum and minimum SGR occurred under IOD by MHL and IDN by FL, respectively (difference 56.34%). The FI of shrimp for the control group did not rank lowest among treatments, confirming that shrimp primarily use scent, not vision, to search for food. FI and FCE of shrimps were both the lowest among treatment groups under IDN by FL and growth was slow, thus FL is not a preferred light source for shrimp culture. Under IOD by MHL, shrimps had the highest FCE and the third highest FI among treatment groups ensuring rapid growth. FAC of shrimp were about 3.31 +/- 0.20 mg/kg. When under IOD by MHL and IDN by FL, FAC was significantly higher than the other treatments (P < 0.05). To summarize, when illuminated by MHL, L. vannamei had not only vivid body color due to high astaxanthin concentration but also rapid growth. Therefore, MHL is an appropriate indoor light source for shrimp super-intensive culture. SGR of shrimp was in significantly negative correlation to FAC of shrimp (P < 0.05). Thus, when FAC increased, SGR did not always follow, suggesting that the purpose of astaxanthin accumulation was not for growth promotion but for protection against intense light. (c) 2005 Elsevier B.V. All rights reserved.

Study on growth factors of intermetallic layer within hot-dipped 25% Al-Zn alloy coating on steel

25%Al-Zn alloy coating performs better than hot dip galvanized coating and 55%Al-Zn-Si coating with regard to general seawater corrosion protection. This study deals with the interfacial intermetallic layer's growth, which affects considerably the corrosion resistance and mechanical properties of 25%Al-Zn alloy coatings, by means of three-factor quadratic regressive orthogonal experiments, The regression equation shows that the intermetallic layer thickness decreases rapidly with increasing content of Si added to the Zn-Al alloy bath, increases with rise in bath temperature and prolonging dip time. The most effective factor that determined the thickness of intermetallic layer was the amount of Si added to Zn-Al alloy bath, while the effect of bath temperature and dip time on the thickness of intermetallic layer were not very obvious.

矮嵩草草甸植被-土壤系统CO2的释放特征

采用静态箱-气相色谱法,对高寒矮嵩草草甸植被-土壤系统CO2释放特征研究结果表明:3个处理(FC、FJ、FL)CO2释放速率具有明显的日变化规律,日最大释放速率出现在13:00左右,最小释放速率在4:00前后,且白天的释放速率均大于夜间;CO2释放速率也具有明显的季节变化特征,植物生长期释放速率明显高于枯黄期,且均表现为正排放;在整个观测期间(6月30日～1月28日)CO2平均释放速率依次为FC>FJ>FL,矮嵩草草甸植物-土壤系统CO2释放速率为438.34±264.12mg/(m2•h)(FC),土壤呼吸速率为313.20±189.74 mg/(m2•h)(FJ),土壤微生物呼吸速率为230.34±145.46mg/(m2•h)(FL),植物根系呼吸占土壤呼吸的26.5%.植物、植物根系以及土壤微生物CO2释放速率与土壤5 cm温度呈极显著正相关关系,相关系数分别为0.858、0.628和0.672(P<0.01).整个系统呼吸、土壤呼吸与土壤5 cm温度可拟和为一指数方程,方程为y=168.03e0.10x86x(R2=0.8783)和y=149.69e0.0745x(R2=0.8189).

Comparative study on CO2 emissions from different types of alpine meadows during grass exuberance period

Potentilla fruticosa scrub, Kobresia humilis meadow and Kobresia tibetica meadow are widely distributed on the Qinghai-Tibet Plateau. During the grass exuberance period from 3 July to 4September, based on close chamber-GC method, a study on CO2 emissions from different treatments was conducted in these meadows at Haibei research station, CAS. Results indicated that mean CO2emission rates from various treatments were 672.09+152.37 mgm-2h-1 for FC (grass treatment); 425.41+191.99 mgrn-2h-1 for FJ (grass exclusion treatment); 280.36+174.83 mgrn-2h-1 for FL (grass and roots exclusion treatment); 838.95+237.02 mgm-2h-1 for GG (scrub+grass treatment); 528.48+205.67 mgm-2h-1for GC (grass treatment); 268.97 ±99.72 mgm-2h-1 for GL (grass and roots exclusion treatment); and 659.20±94.83 mgm-2h-1 for LC (grass treatment), respectively (FC, FJ, FL, GG, GC, GL, LC were the Chinese abbreviation for various treatments). Furthermore, Kobresia humilis meadow, Potentilla fruticosa scrub meadow and Kobresia tibetica meadow differed greatly in average CO2 emission rate of soil-plant system, in the order of GG＞FC＞LC＞GC. Moreover, in Kobresia humilis meadow,heterotrophic and autotrophic respiration accounted for 42% and 58% of the total respiration of soil-plant system respectively, whereas, in Potentilla fruticosa scrub meadow, heterotrophic and autotrophic respiration accounted for 32% and 68% of total system respiration from G-G; 49% and 51%from GC. In addition, root respiration from Kobresia humilis meadow approximated 145 mgCO2m-2h-1,contributed 34% to soil respiration. During the experiment period, Kobresia humilis meadow and Potentilla fruticosa scrub meadow had a net carbon fixation of 111.11 grn-2 and 243.89 grn-2,respectively. Results also showed that soil temperature was the main factor which influenced CO2 emission from alpine meadow ecosystem, significant correlations were found between soil temperature at 5 cm depth and CO2 emission from GG, GC, FC and FJ treatments. In addition, soil moisture may be the inhibitory factor of CO2 emission from Kobresia tibetica meadow, and more detailed analyses should be done in further research.

Determination of amines using 2-(11H-benzo[a]carbazol-11-yl) ethyl chloroformate (BCEC-Cl) as labeling reagent by HPLC with fluorescence detection and identification with APCI/MS

A pre-column derivatization method for the sensitive determination of amines using a labeling reagent 2-(11H-benzo[a]-carbazol-11-yl) ethyl chloroformate (BCEC-Cl) followed by high-performance, liquid chromatography with fluorescence detection has been developed. Identification of derivatives was carried out by LC/APCI/MS in positive-ion mode. The chromophore of 1,2-benzo-3,4-dihydrocarbazole-9-ethyl chloroformate (BCEOC-Cl) reagent was replaced by 2-(11H-benzo[a]-carbazol-11-yl) ethyl functional group, which resulted in a sensitive fluorescence derivatizing reagent BCEC-Cl. BCEC-Cl could easily and quickly label amines. Derivatives were stable enough to be efficiently analyzed by HPLC and showed an intense protonated molecular ion corresponding m/z [M+ H](+) under APCI/MS in positive-ion mode. The collision-induced dissociation of the protonated molecular ion formed characteristic fragment ions at m/z 261.8 and m/z 243.8 corresponding to the cleavages of CH2O-CO and CH2-OCO bonds. Studies on derivatization demonstrated excellent derivative yields over the pH 9.0-10.0. Maximal yields close to 100% were observed with three- to four-fold molar reagent excess. In addition, the detection responses for BCEC-derivatives were compared to those obtained using 1,2-benzo-3,4-dihydrocarbazole-9-ethyl chloroformate (BCEOC-Cl) and 9-fluorenyl methylchloroformate, (FMOC-Cl) as labeling reagents. The ratios I-BCEC/I-BCEOC = 1.94-2.17 and I-BCEC/I-FMOC = 1.04-2.19 for fluorescent (FL) responses (here, I was relative fluorescence intensity). Separation of the derivatized amines had been optimized on reversed-phase Eclipse XDB-C-8 column. Detection limits calculated from 0.50 pmol injection, at a signal-to-noise ratio of 3, were 1.77-14.4 fmol. The relative standard deviations for within-day determination (n = 11) were 1.84-2.89% for the tested amines. The mean intra- and inter-assay precision for all amines levels were < 3.64% and 2.52%, respectively. The mean recoveries ranged from 96.6% to 107.1% with their standard deviations in the range of 0.8-2.7. Excellent linear responses were observed with coefficients of > 0.9996. (C) 2006 Elsevier B.V. All rights reserved.

Blood-gas properties of plateau zokor (Myospalax baileyi)

Plateau zokor (Myospalax baileyi) is one of the blind subterranean mole rats that spend their life solely underground in scaled burrows. It is one of the special species of the Qinghai-Tibet plateau. In their burrows, oxygen is low and carbon dioxide is high and their contents fluctuate with the change of seasons, soil types, rain and depth of burrows. However, plateau zokors show successful adaptation to that extreme environment. In this study, their adapting mechanisms to the hypoxic hypercapnic environment were analyzed through the comparison of their blood-gas properties with that of pikas (Ochotona curzniae) and Sprague-Dawley rats. The results indicated that plateau zokors had higher red blood corpuscle counts (8.11 +/- 0.59 (10(12)/L)) and hemoglobin concentrations (147 +/- 9.85 g/L), but hematocrit (45.9 +/- 3.29%) and mean corpuscular volume (56.67 +/- 2.57 fL) were lower than the other rodents. Their arterial blood and venous blood pH were 7.46 +/- 0.07 and 7.27 +/- 0.07. Oxygen pressure in arterial blood of plateau zokors was about 1.5 times higher than that of pikas and rats, and it was 0.36 and 0.26 times in their venous blood. Partial pressure for carbon dioxide in arterial and venous blood of plateau zokors was 1.5-fold and 2.0-fold higher, respectively, than in rats and pikas. Oxygen saturation of plateau zokors was 5.7 and 9.3 times lower in venous blood than that of pikas and rats, respectively. As result, the difference of oxygen saturation in arterial blood to venous blood was 2- and 4.5-fold higher in plateau zokors as that of pikas and rats, respectively. In conclusion, plateau zokors had a high tolerance to pH changes in tissues, together with strong capabilities to obtain oxygen from their hypoxic-hypercapnic environment. (c) 2006 Published by Elsevier Inc.

Pre-column derivatization of amines with 1,2-benzo-3,4-dihydrocarbazole-9-isopropyl chloroformate followed by LC-fluorescence and LC-APCI-MS

A pre-column derivatization method for the sensitive determination of amines using the labeling reagent 1,2-benzo-3,4-dihydrocarbazole-9-isopropyl chloroformate (BCIC-Cl) followed by high-performance liquid chromatography with fluorescence detection has been developed. Identification of derivatives is carried out by high performance liquid chromatography/atmospheric pressure chemical ionization (LC-APCl-MS-MS). The chromophore of 2-(9-carbazole)-ethyl chloroformate (CEOC) reagent is replaced by 1,2-benzo-3,4-dihydrocarbazole-9-isopropyl functional group, which results in a sensitive fluorescence derivatizing reagent BCIC-Cl. BCIC-Cl can easily and quickly label amines. Derivatives are stable enough to be efficiently analyzed by high-performance liquid chromatography and show an intense protonated molecular ion corresponding m/z [MH](+) under APCl in positive-ion mode. The collision-induced dissociation of protonated molecular ion formed a product at m/z 260 corresponding to the cleavage of CH2-OCO bond. Studies on derivatization demonstrate excellent derivative yields over the pH 9.0-10.0. Maximal yields close to 100% are observed with a 3 to 4-fold molar reagent excess. In addition, the detection responses for BCIC derivatives are compared with those obtained using CEOC and FMOC as derivatization reagents. The ratios of l(BCIC)/l(CEOC) and l(BCIC)/l(FMOC) are, respectively, 1.23-3.14 and 1.25-3.08 for fluorescent (FL) responses (here, l is relative fluorescence intensity). Separation of the derivatized amines had been optimized on reversed-phase Eclipse XDB-C-8 column. Detection limits are calculated from 1.0 pmol injection, at a signal-to-noise ratio of 3, are 10.6-37.8 fmol. The mean interday accuracy ranges from 94 to 105% for fluorescence detection with the largest mean %CV < 7.5. The mean interday precision for all standards is < 6.0% of the expected concentration. Excellent linear responses are observed with coefficients of > 0.9997.