丙烯腈、1-十六烯和发烟硫酸反应产物的分析和表征

利用丙烯腈、1-十六烯和发烟硫酸的反应合成了新型丙烯酰胺基双亲单体,本文首次给出了该化合物的红外光谱、质谱和~1H-NMR 谱图,元素分析,不饱和度和酸值测定,红外光谱、质谱和~1H-NMR 谱等实验结果说明,所得化合物为2-丙烯酰胺基十六烷磺酸(AMC_(16)S),AMC_(16)S 不但带有可聚合双键,而且具有良好的胶体化学性质,其水溶液的临界胶束浓度为8.0×10~(-5)g/ml,1(重量)％浓度溶液的表面张力为33.5mN/m。

系列人参皂甙的快原子轰击质谱研究

本文研究了九种人参皂甙的正离子快原子轰击质谱(PFAB)及负离子快原子轰击质谱(NFAB)。鉴于某些皂甙类及其它生物活性物质含量少、分离难等特点,工作中我们重点放在提高灵敏度上,并选择了适合于皂甙的底物,用0.5μg的样品,得到了满意的谱图。

超高灵敏显色反应及其应用的研究——Ⅳ.生命活性元素锌的高灵敏光度法

本文研究了Zn~(2+)-SCN~--RB~+-PVA_(124)超高灵敏显色体系及锌的分光光度测定条件。缔合物的吸收峰在607nm处,表观摩尔吸光系数达2.6×10~6L·mol~(-1)·cm~(-1)。Zn~(2+)浓度在0～0.5μg/25ml范围内符合比耳定律。测得缔合物的组成比Zn~(2+):RB~(+)为1:30。初步研讨了显色反应的机理。方法有较好的选择性,可不经预分离,成功地用于水样,麦饭石浸出液、花粉和发样中痕量锌的测定。

超高灵敏显色反应及其应用的研究——Ⅴ.钨(Ⅴ)-SCN~--乙基罗丹明B-PVA-124体系

本文研究了在聚乙烯醇-124存在下,钨(V)-SCN~--乙基罗丹明B超高灵敏显色体系。缔合物λ_(max)=585nm,表观摩尔吸光系数ε_(585)=1.9×10~6L·mol~(-1)·cm~(-1)。钨浓度在0.1～1.5μg/25ml范围内符合比尔定律,方法曾用于水样及钢样中痕量钨的测定,结果较满意。

超高灵敏显色反应及其应用的研究Ⅲ.锑(Ⅲ)—乙基罗丹明B—碘化物体系

本文研究了在聚乙烯醇-124存在下,锑(Ⅲ)—乙基罗丹明B—碘化物的显色反应。缔合物λ_(max)=605nm表观摩尔吸光系数为7.0×10~3。锑浓度在0.1～2.5μg/25ml时服从比尔定律。藉氢化物分离技术,本法可用于地球化学标样中痕量锑的测定。

超高灵敏显色反应及其应用的研究 Ⅱ.汞(Ⅱ)-乙基罗丹明 B-碘化物体系

本文研究了在聚乙烯醇-124存在下,汞(Ⅱ)-乙基罗丹明B_碘化物超高灵敏显色体系及其反应机理。缔合物λ_(max)=605nm,表观摩尔吸光系数ε_(605nm)=1.14×10~6L.mol~(-1).cm~(-1)。汞(Ⅱ)浓度在0—2.5μg/25ml时符合比耳定律。方法可用于水样及地球化学标样中痕量汞的测定。

Pilot-scale fermentation and purification of the recombinant allophycocyanin over-expressed in Escherichia coli

A recombinant allophycocyanin (rAPC), used for treatment of tumors, has been expressed in E. coli which was grown in glucose fed-batch culture in a 30 l fermentor. Recombinant allophycocyanin was purified from soluble E. coli cell lysate using hydrophobic interaction chromatography followed by chromatography using amylose affinity column. The purity of product was greater than 98% and yielded an average of 5.5 g kg(-1) dry cells. Recombinant allophycocyanin significantly inhibited H-22 hepatoma (p (0.01) in mice with inhibition rates ranging from 36% to 62% with doses from 6.25 to 50 mg kg(-1) d(-1).

胶州湾超微型浮游真细菌16s rDNA RFLP分析相关技术的研究

通过生态学与分子生物学相结合的方法，对胶州湾超微型浮游真细菌从16s rDNA的角度分析其多样性，建立了一系列行之有效的方法，对于今后开展特定的超微型生物的研究提供他有益的借鉴。主要结果如下：1.建立了行之有效的从极稀密度的海水中提取浮游细菌总基因组DNA的方法，其得率约为0.3～0.5 μg/L海水，并且提供了DNA浓缩、纯化的一系列步骤，最后所制得的DNA其纯度和含量足以开展后续工作。2.建立了可靠的具有较高转化效率的感受态细胞，并从多个途径来制备，并对各种方法的成败及注意事项进行了探讨。3.对PCR反应的条件进行了深入的探讨，成功在从混合型基因组中扩增出细菌特异性引物(Universal 1406R和Eubacterial 68F)所对应的16s rDNA。并且对PCR的几个控制因子进行了分析，对于今后采用不同引物的高效扩增将提供有益的帮助。4.深入地对TA克隆效率进行了探讨，较好地将PCR混合产物分离开，并成功地导入大肠杆菌DH5α菌株，并对重组质粒进行了初步的RFLP分析，为今后通过序列测定构建系统进化树奠定了基础。5.RFLP分析表明胶州湾海水中至少含有7种亲缘关系较远的真细菌。

利用两种大型海藻制备生物能源的探索研究

生物质燃料乙醇和沼气都属于发展潜力巨大的生物质能源，大力发展生物质燃料乙醇和沼气对解决当今能源危机、环境污染问题和促进我国经济发展具有重要意义。 本文就海带化工废弃物——海带渣糖化技术、浒苔糖化技术及初步酒精发酵技术、海带沼气发酵技术、浒苔沼气发酵技术进行了可行性研究。 1、海带渣总糖含量为52.6%，海带渣总糖中葡萄糖含量占90.9%，另外还有少量的半乳糖、甘露糖和木糖，这说明海带渣是非常优良的能源生物质。 2、海带渣糖化工艺采取稀硫酸预处理后纤维素酶酶解产糖工艺。海带渣最佳稀硫酸预处理条件为预处理温度121℃、硫酸浓度0.6%、预处理时间60 min，此时海带渣纤维素酶解产糖可达187.8 mg/g干藻。 3、优化了预处理海带渣纤维素酶酶解产糖工艺。各因素对海带渣酶解产糖的影响依次为：pH>温度>时间>酶用量，海带渣纤维素酶解产糖的最佳条件为温度45℃、pH5.2、时间48小时、酶用量16 mg/g干藻，此时糖产量为238.9 mg/g干藻。 4、浒苔总糖含量为67.2%，浒苔总糖主要有葡萄糖、木糖、葡萄糖醛酸和鼠李糖组成，其中葡萄糖和木糖可以做为酒精发酵的原料，这两种糖占总糖含量的51%。 5、以鲜浒苔为原料研究了浒苔稀硫酸水解工艺和纤维素酶酶解工艺。发现浒苔酸水解产糖效果明显优于纤维素酶酶解。 6、以干浒苔为原料研究了浒苔酸解工艺。硫酸水解干浒苔产糖能力优于盐酸、磷酸和马来酸，水解时间为60 min、硫酸浓度为1.8%的时候可用于酒精发酵的糖（葡萄糖和木糖）总产量达到最大值为230.5 mg/g干浒苔，占此条件下总还原糖产量的48.6%。同时发现干浒苔比鲜浒苔更易被硫酸水解产糖。 7、初步研究了浒苔酒精发酵工艺。初步工艺中酒精产量较低、测得酒精在发酵液中浓度为0.23%（v/v），后续工作中需要对酒精发酵工艺进行优化。 8、海带与牛粪比例为4:1（w/w）是海带与牛粪联合厌氧消化的最佳比例，此时产气时间最长，达到37天，总产气量最高，达到13600 mL。 9、在海带与牛粪联合厌氧消化中，增加接种量到15 g（干重）时发酵周期为39天，总产气量为14630 mL，TS产气量为152.4 mL/gTS，接种量为21 g（干重）时发酵周期为36天，总产气量为14090 mL，TS产气量为138.1 mL/gTS，可见适当增加接种量可促进产气量的增加。 10、浒苔与牛粪比例为4:2（w/w）是浒苔与牛粪联合厌氧消化的最佳比例，此时产气时间为33天，总产气量最高达到6785 mL。 11、在浒苔与牛粪联合厌氧消化中，增加接种量到7.5 g（干重）时发酵周期为38天，总产气量为7470 mL，TS产气量为155.6 mL/gTS，接种量为10.5 g（干重）时发酵周期也为38天，总产气量为7020 mL，TS产气量为137.6 mL/gTS，可见增加接种量可促进产气量的增加。

孔石莼主要化学组分的提取分析及多糖生物活性的研究

孔石莼（Ulva pertusa Kjellm）是一种大型经济海藻，除了可直接食用外在医药方面也有广泛的应用。在古代医书上便记载有多种药用活性，我国民间也流传用孔石莼治疗中暑、水肿和小便不利等病症。本论文比较全面地分析了孔石莼多糖和类脂等主要化学组合，探讨用红外和核磁光谱来解释多糖结构，并首次研究了孔石莼多糖的体外抗肿瘤的体内降血脂活性。为进一步开发孔石莼资源做了部分基础性的工作。通过对提取扎石莼多糖和类脂的方法研究，确定了两种主要组仇的提取方法，得到了多糖样品WPS、APS、DWPS、DAPS和类脂样品。用化学和仪器分析方法对多糖样品进行了水分、灰分、硫酸基含量、总糖含量、特性粘度、元素分析、单糖分析、氨基酸分析、红外和核磁光谱分析等。结果表明：用乙醇脱脂后的脱脂孔石莼水提物（DWPS）得率最高，含硫酸基量和总糖量最高，特性粘度最小；孔石莼水提物（WPS）得率和含硫酸基量次之；孔石莼碱提物（APS）因深解性不好未能按常规方法测定其总糖含量，且未检测出水解硫酸根。孔石莼多糖的单糖要有鼠李糖、葡萄糖、木糖、糖醛酸等，并检测到少量的甘露糖和阿拉伯糖。谷氨酸、天冬西氨酸是多糖样品蛋白质中的主要氨基酸，D－3－OH－半胱氨酸含量亦较高。IR谱图中出现了硫酸基的吸收峰：~H－NMR及~(13)C－NMR谱图中出现了鼠李糖和糖醛酸的特征峰。类脂样品中，非极性类脂与极性类脂的比例约为1：2；GC－MS 分析结果表明，不饱和脂质含量达61.37％，其中多不饱和脂类占44.17％，n-3系列高度不饱和脂类占39.90％。三个多糖样品WPS、DWPS、DAPS进行抗口腔上皮癌（KB）和肝癌（Bel）肿瘤细胞体外实验，结果表明在5μg/ml的剂量时三个样品抑制率均在10％以下，无明显活性表现。以烟酸肌醇为阳性对照药，蒸馏水为空白对照，对孔石莼水提多糖WPS进行了三个剂量的降血脂动物实验。实验结果表明：脂蛋白改善情况有明显的剂量依赖性；中高剂量组可显著降低胆固醇、甘油酸三脂，明显改善脂蛋白胆固醇的含量分布：高剂量样品组（500mg/kg·d~(-1)）的药用活笥最高，与相同剂量的阳性对照药烟酸肌醇结果相比，药效理佳。并对孔石莼多糖血脂的机理进行了讨论。

GUS基因在优化海带表达系统中的应用

本文研究了如何应用GUS基因优化海带表达系统。首先研究了GUS在各种海带材料中的本底值，而后以GUS基因作为报告基因比较了几种基因工程常用启动子在海带中启动瞬间表达的效率。选择表达效率较高的真核藻类启动子与GUS基因融合，在海带中获得了稳定表达的结果。分别用组织化学染色法和荧光分析法对海带雌配子体、雄配子体、孤雌生殖海带幼孢子体和二倍体海带孢子体等四种不同的材料的GUS本底进行了检测到GUS本底。荧光分析法所得数据显示，这四种材料的GUS活性分别为3.4±0.3、5.2±0.8、14.4±1.8、12.5±1.4 (pmol MU mg~(-1) protein min~(-1))，与高等植物类似，说明GUS基因可作为报造基因用于海带基因工程研究。选用真核藻启动子FCP启动子、高等植物基因工程中常用的高效Ubi启动子和CaMV35S启动子，与GUS基因融合构建四种质粒，通过基因松的方法转化孤雌生殖海带幼孢子体，比较GUS基因在海带中的瞬间表达量。实验结果表明，与CaMV35S和FCP融合的GUS基因在海带中表达效率较高。采用基因松的方法用FCP启动子－GUS基因转化海带雌配子体，通过诱导孤雌生殖获得孤雌生殖海带幼孢子体，用组织染色法检测到了GUS活性，并从海带的总DNA扩增到了FCP启动子－GUS基因的特征条带，从而提示了FCP启动子能引导外源基因在海带中稳定表达。为了找寻高效筛选元件进行孤雌生殖海带对草丁膦的敏感性实验，用统计学的方法得出了草丁膦对不同长度孤雌海带的半致死剂量。研究发现海带全长0.5-1.6cm范围内，草丁膦的LD_(50)与海带长度不相关，同时发现浓度在2-5μg/ml狭小范围内的草丁膦对孤雌海带比高浓度具有更为明显的毒性反应。本文结果提示草丁膦抗性基因－bar基因有可能成为海带基因工程更为理想的选择标记，因为海带对草丁膦比对氯霉素和潮霉素更敏感，而后两者是目前采用的筛选压力。本文结果为海带表达系统的优化，提供了有效的报告基因、启动子和选择标记元件。

Effects of body size and water temperature on food consumption and growth in the sea cucumber Apostichopus japonicus (Selenka) with special reference to aestivation

To investigate the effects of body size and water temperature on feeding and growth in the sea cucumber Apostichopus japonicus (Selenka), the maximum rate of food consumption in terms of energy (C-maxe; J day(-1)) and the specific growth rate in terms of energy (SGRe; % day(-1)) in animals of three body sizes (mean +/- SE) - large (134.0 +/- 3.5 g), medium (73.6 +/- 2.2 g) and small (36.5 +/- 1.2 g) - were determined at water temperatures of 10, 15, 20, 25 and 30 degrees C. Maximum rate of food consumption in terms of energy increased and SGRe decreased with increasing body weight at 10, 15 and 20 degrees C. This trend, however, was not apparent at 25 and 30 degrees C, which could be influenced by aestivation. High water temperatures (above 20 degrees C) were disadvantageous to feeding and growth of this animal; SGRe of A. japonicus during aestivation was negative. The optimum temperatures for food consumption and for growth were similar and were between 14 and 15 degrees C, and body size seemed to have a slight effect on the optimal temperature for food consumption or growth. Because aestivation of A. japonicus was temperature dependent, the present paper also documented the threshold temperatures to aestivation as indicated by feeding cessation. Deduced from daily food consumption of individuals, the threshold temperature to aestivation for large and medium animals (73.3-139.3 g) was 24.5-25.5 degrees C, while that for small animals (28.9-40.7 g) was between 25.5 and 30.5 degrees C. These values are higher than previous reports; differences in sign of aestivation, experimental condition and dwelling district of test animals could be the reasons.

Pretreatment Method for Nitrogen Isotopic Analysis of Nitrate in Seawater

A suitable method for the pretreutment of dissolved nitrate samples in seawaters for nitrogen isotopic analysis was established. First, the seawater samples were processed by removing nitrite and amonium. Then Devard's alloy was added in sample for conversion of dissolved nitrate to ammonium. The sample was distilled, and then the ammonium condensate was collected with zeolite. after distillation, the collected condensate was filtered and prepared for determining nitropic values. Some tests of the method were conducted. The distillation condition, the influence of salinity on nitrogen isotopic analysis, absorption of ammonium onto zeolite and an improved method on a large volume of seawater were discussed in this study. The results showed that the distillation step had an average recovery of (104.9 +/- 4.2) % (n = 6) when distillating every 300 mL aliquot of the sample under a strong alkaline condition with 0.5 g devard's alloy and a distillation time of 30 min. The nitrogen isotopic fractionation decreased markedly when salinity was increased from 0% to 0.5%; further increase(1% - 3.5%) showed little effect. The adsorption rate of ammonium onto zeolite had a high yield of (95.96 +/- 1.08) % (n = 6) in average. An improved collection method was used to process a large volume of seawater with several distillations, and had good effect on analysis. The method had been applied to analyze water samples collected from Changjiang estuary. The analytical results indicate that the method is suitable for delta N-15 analysis of dissolved nitrate in seawaters. The present method could provide valuable information about the source and cycle mechanism of dissolved nitrogen in estuary waters.

C, N and P regeneration by a detritivorous fish, Liza haematocheila T. & S.: effects of temperature, diet and body size

Detritus, as a nutrients reservoir, affects the trophic structure and dynamics of communities and supports a greater diversity of species and longer food chains. Detritivorous fish is an important organism to regenerate the nutrients from sediments. Despite the numerous studies on the nutrients cycle in fish, only a few attempts have been made to quantify the regenerating ability. In the present study, we chose the common detritivorous fish redeye mullet as the research object. Redeye mullet is also a common poly-culture fish in China. Diet, including a commercial diet mostly used in aquaculture and a home-made diet with contents close to detritus, was used and considered as a fixed factor. Temperature was also considered as a fixed factor as much research has shown that temperature has significant effects on fish metabolism. Moreover, body size was regarded as a covariate under analysis of covariance. Three key nutrients, namely carbon, nitrogen and phosphorus, were used to measure the nutrient-regenerating ability of redeye mullet under laboratory conditions. The results showed that the nutrient regeneration in percent of the consumption decreased with increasing temperature. Carbon and nitrogen regeneration of redeye mullet fed on commercial diet was lower than those of the home-made diet group, while the opposite was found for phosphorus. In each group, the amount of regenerated nutrients increased linearly with body size. Fed on the home-made diet, 5-g fish at 25 degrees C can regenerate 210.822 mg C, 37.533 mg N and 0.727 mg P per day.

毛细管电泳法分析唐古特白刺种子中两种生物碱

利用毛细管电泳法分离测定唐古特白刺种子中的尿囊素和吲哚生物碱1-methyl-1，2，3，4-tetrahydro-β-earboline-3-carboxylic acid（MTCCA），所用毛细管规格为48，5cm×50μm i．d．，DAD检测波长220nm，最佳分离条件：电压19kV，分离温度25℃，背景电解质为含有32mmol／LSDS，体积分数10．0％乙腈的32mmol／L硼酸溶液，pH10．0。MTCCA与尿囊素分别在350．0～11.0μg／mL和112．5～3．5μg／mL质量浓度范围内与电泳峰面积呈现良好线性关系，检出限分别为5．0μg/mL和2.5μg／mL。对标准品进行6次测定，迁移时间的RSD为1.1％和1．4％，峰面积的RSD为2.3％和0.82％。