结核分枝杆菌基因组学及特异检测技术研究

结核分枝杆菌(M.tuberculosis)，俗称结核杆菌，是引起结核病的病原菌。可侵犯全身各器官，但以肺结核为最多见。结核病至今仍为重要的传染病。估计世界人口中1/3感染结核分枝杆菌。据WHO报道，每年约有800万新病例发生，至少有300万人死于该病。我国建国前死亡率达200-300人/10万，居各种疾病死亡原因之首，建国后，随着人民生活水平提高，卫生状态改善，特别是开展了群防群治，儿童普遍接种卡介苗，结核病的发病率和死亡率大为降低。由于耐药结核的出现已及HIV共感染等因素影响，目前发病率又有上升趋势。对于结核病的防治而言，其首要问题是结核病的诊断，而结核病的实验室检查已成为临床医生诊断结核病、判断疗效和评估预后不可缺少的手段。结核病的临床诊断主要根据病史、痰培养、涂片抗酸染色、胸片等结果，其中痰结核检查是诊断肺结核的重要依据之一，检测方法常规采用细菌学检查。痰中细菌学检查主要采用痰涂片和培养方法，涂片虽简单易行，但阳性率低，培养虽为金标准，但周期太长，均难以满足临床需要。细菌对部分患者可造成误诊或漏诊。近年来随着现代科学技术的不断进展，分子生物学技术在结核病诊断方面取得了显著的成绩，实时动态荧光定量(FQ—PCR)技术又是在原有的PCR技术上一次质的飞跃。 尽管许多文献已经报道了结核耐药的分子机理，但耐药的分子机理并没有充分了解。本研究利用全基因组鸟枪法测序和高通量测序技术，测定了两株结核分枝杆菌菌株TFJA和TFJB全基因组序列，分别为药物敏感株和耐多药株。在序列初步拼接后，获得了由189个DNA contigs（DNA序列连接群）组成的基因组框架图。借助10kb左右插入片段克隆的正反向末端序列信息以及PCR扩增技术，确定了全部DNA contigs在基因组上的物理位置，最后补上了所有“间隙(gap)” ，得到了全基因组序列。三种限制性酶切物理图谱证明了序列拼接的正确性。两株结核菌基因组大小为4.4Mbp左右，GC含量都在65.6%左右，其中TFJB株比TFJA株大7kbp，分别具有3591、3648个基因，83.7%的基因位于先导链上。我们比较了两株细菌的949SNPs位点，发现94个在TFJB中不同nsSNPs，这些SNPs位点的碱基在其它的结核分枝杆菌基因组中是相同的。我们去掉了那些代谢途径与毒力代谢无关的基因，最后选择了6个基因的18个SNP，检测这些SNP在277株耐药或者药物敏感结核分枝杆菌中的保守性以及其耐药相关性。用PCR扩增含有SNP的序列，用统计学检验这些SNP是否耐药相关，结果发现，TFJB0109_CDS04405基因的407位点SNP（g->a）和TFJB0109_CDS00149基因上的SNP（720a->c，722插入t，727t->g，729g->t，以及735g->t）可能与SM耐药相关，其P-value分别是0.04128、0.02160和0.01394（见表3）。 结核是一种慢性传染病，其病原体为TB。在全基因组序列分析研究的同时，我们筛选到一套灵敏度较高、特异性较好的REAL TIME-PCR检测引物，并用它对105份不同发病时间的病人标本进行了检测。初步结果表明，应用本方法，样本检出率为98%，这一结果比涂片及分离培养更灵敏。

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。

黄海浮游动物功能群的研究

浮游动物在海洋生态系统物质循环和能量流动中起着至关重要的作用。浮游动物物种组成、生物量和次级生产力的变化会改变生态系统的结构和功能。在黄海生态系统中如何描述这个过程，并使它易于模拟是本论文的研究目的。生物量和生产力是海洋生态系统食物网的基础。谁是浮游动物生物量和次级生产力的基础？哪些种类在生态系统中起关键作用？这些问题在黄海这样的温带陆架边缘海区很难回答，原因是物种组成、生物量和生产力的季节变化显著。因此，在对黄海食物产出的关键过程进行模拟时，需要应用既准确又简便的方法来对浮游动物群落的生态过程进行模拟。在对黄海浮游动物群落结构和物理海洋学特征进行充分的分析之后，浮游动物功能群的方法被确定用来进行黄海生态系统结构和功能的模拟。 根据浮游动物的粒径、摄食习性和营养功能，黄海浮游动物被分为6个功能群：大型浮游甲壳动物功能群（Giant crustacean，GC）、大型桡足类功能群（Large copepods, LC）、小型桡足类功能群（Small copepods，SC）、毛颚类功能群（Chaetognaths）、水母类功能群（Medusae）和海樽类功能群（Salps）。GC、LC和SC是按照粒径大小而划分的功能群，他们是高营养层次的主要食物资源。毛颚类和水母类是两类胶质性的肉食性浮游动物功能群，他们与高营养层次竞争摄食饵料浮游动物；海樽类与其他浮游动物种类竞争摄食浮游植物，而本身的物质和能量却不能有效的传递到高营养层次。本文研究报道了浮游动物各功能群的时空分布、基于浮游动物动能群的黄海生态区划分、饵料浮游动物功能群的生产力、毛颚类对浮游动物的摄食压力以及中华哲水蚤（Calanus sinicus）的摄食生态学。 春季，浮游动物生物量为2.1 g m–2，GC、LC和SC对生物量的贡献率分别为19, 44 和 26%。高生物量的LC和SC功能群主要分布于山东半岛南岸的近岸海域，而GC主要分布在远岸站位。夏季，浮游动物的生物量为3.1 g m–2，GC贡献了73%。GC、LC和SC主要分布在黄海的中部海域。秋季，浮游动物生物量为1.8 g m–2，GC、LC和SC的贡献率相似，分别为36, 33和23%，高生物量的GC和LC分布在黄海中部，而SC主要分布在远岸站位。GC和LC是冬季浮游动物生物量（2.9 g m–2）的优势功能群，分别贡献率了57%和27%，高生物量的GC、LC和SC都分布在黄海的中部海域。与GC、LC和SC相比，毛颚类生物量较低，主要分布于黄海的中北部海域。水母类（本文中指小型水母类）和海樽类斑块分布明显，主要分布于黄海沿岸和北部海域。属于不同功能群的约10个种类为浮游动物的优势种，控制着浮游动物群落的动态。 春季，黄海可以被分成4个浮游动物生态区，浮游动物生物量的分布中心位于山东半岛南岸近岸海域，与第一个生态区相对应，LC和SC在分布中心起主要的控制作用；夏、秋和冬季，黄海分别被分成3、4和3个生态区，浮游动物生物量的分布中心均位于黄海的中部海域，均与各季节的第一个生态区相对应，GC和LC是分布中心生态区的优势功能群，对分布中心起主要的控制作用。黄海冷水团（YSCBW）在GC、LC和SC的空间分布模式中起着重要的作用。黄海不同季节浮游动物生态区的空间分布模式及生态区中起控制作用的优势功能群类别有着重要的生态学意义。 我们将饵料浮游动物功能群细化为0.16–0.25 mm、0.25–0.5 mm、0.5–1 mm、1–2 mm和 >2 mm5个粒径组。应用生物能量学的方法研究了不同粒径浮游动物的生产力。结果表明：浮游动物次级生产力5月份最高，为91.9 mg C m–2 d–1，其次是6月和9月，分别为75.6 mg C m–2 d–1和65.5 mg C m–2 d–1，8月、3月和12月较低，仅为42.3 mg C m–2 d–1、35.9 mg C m–2 d–1和27.9 mg C m–2 d–1。根据这些结果，黄海浮游动物年次级生产力为18.9 g C m–2 year–1。0.16–0.25 mm和 0.25–0.5 mm 两个粒径组对浮游动物次级生产力的贡献率为58–79%，即相对应的SC功能群的周转率（P/B, 0.091–0.193 d–1）要高于GC和LC。 黄海毛颚类功能群的优势种类为强壮箭虫（Sagitta crassa）、纳嘎箭虫（S. nagae）、肥胖箭虫（S. enflata）和百陶箭虫（S. bedoti）。我们对这四种箭虫的生产力和对浮游动物生物量和生产力的摄食压力进行了研究。结果表明：黄海毛颚类总的生物量为98–217 mg m–2，总的生产力为1.22–2.36 mg C m–2 d–1。黄海毛颚类的生物量占浮游动物总生物量的6.35–14.47%，而生产力仅占浮游动物总生产力的2.54–6.04%。强壮箭虫和纳嘎箭虫是黄海毛颚类功能群的绝对优势种，控制着黄海毛颚类群落的动态。黄海毛颚类总的摄食率为4.24–8.18 mg C m–2d–1，对浮游动物现存量和生产力总的摄食压力分别为为0.94%和12.56%。黄海冬季，浮游动物的现存量和生产力为0.4 g C m–2和0.026 g C m–2d–1，而毛颚类的摄食压力却达到了全年的最大值，为1.4%和20.94%。因此，毛颚类的摄食可能对冬季浮游动物群落结构造成重要的影响。通过不同体长组箭虫的摄食率可以推断，黄海毛颚类全年主要摄食小型桡足类，对SC功能群的摄食压力最大。但是在夏季黄海冷水团形成的月份，毛颚类对前体长为2 mm的LC功能群中的种类摄食压力也较大，但此时，由于优势种中华哲水蚤进入滞育阶段，因此毛颚类的摄食会对其种群数量造成严重的影响。 中华哲水蚤在春、秋季的摄食率分别为2.08–11.46和0.26–3.70 µg C female–1 day–1，与微型浮游生物的现存量呈显著的正相关。春季，在黄海的北部，中华哲水蚤通过摄食微型浮游生物吸收的碳量能够满足其代谢和繁殖需求，而在黄海的南部和秋季黄海冷水团锋区附近，中华哲水蚤必须通过摄食其他类型的食物资源来维持其代谢和生殖需求。较低的摄食率、无产卵以及种群中CV期桡足幼体占优势表明，秋季中华哲水蚤在黄海冷水团区域内处于滞育状态。中华哲水蚤优先摄食微型原生动物，并且春季中华哲水蚤总的生长效率（GGE, 3–39%）与食物中微型原生动物的比例呈显著的正相关，表明微型原生动物具有较高的营养价值。但是，因较低的产卵率（0.16–12.6 eggs female–1 day–1）而导致的中华哲水蚤较低的总生长效率（13.4%），可能就是由于其食物中的必需营养成分含量不足（或缺乏）造成的。 本文从生物量的角度，对黄海浮游动物各功能群的时空分布、生态区划分进行了研究报道，对GC、LC和SC功能群的生产力、毛颚类对浮游动物的摄食压力和中华哲水蚤的摄食生态学进行了较为深入的研究，这些结果为黄海食物产出的关键过程的模拟提供了基础资料。今后的研究重点应搞清楚黄海水母类对浮游动物次级生产力的摄食压力和海樽类在食物产出模型中产生的负效应的程度，浮游动物各功能群的组成、季节变化和空间分布模式的长期变化，尤其是在气候变化和人类活动的影响下，将是今后研究的重点。

两种红树林植物海漆和海桑化学成分研究

本论文对两种红树林植物海漆 (Excoecaria agallocha L.) 和海桑（Sonneratia caseolaris L.）的化学成分进行了系统研究。 采用常规的硅胶柱层析、制备薄层层析、凝胶 Sephadex LH-20 柱层析、MCI柱层析、反相硅胶柱层析、半制备型 HPLC 以及重结晶等手段，从海漆 (Excoecaria agalloch L. ) 中分离得到 40 个化合物，从海桑（Sonneratia caseolaris L.）中分离得到 30 个化合物。利用各种现代波谱技术 (IR、UV、ESI-MS、EI-MS、1D-NMR、2D-NMR等) 及其化学物理性质，确定了海漆中 32 个化合物的结构，其中包括 1 个新的三萜天然产物以及 15 个首次从海漆中报道的化合物；确定了海桑中 27 个化合物的结构，其中包括 1 个新化合物和一个首次报道其碳谱数据的化合物。本文为首次报道海桑的化学成分研究。 对海漆和海桑粗提物及分离得到的部分化合物进行了抗肝癌细胞毒活性筛选，化合物S22表现出较强活性，其IC50为2.8 μg/mL；海漆和海桑粗提物及其它部分单体化合物只表现出微弱活性；阳性对照丝裂霉素C的IC50为1.1 μg/mL。 对分离得到的部分样品还进行了抗菌活性测试，各样品在测试浓度下对测试菌均未表现出明显的抗菌活性。 首次研究了海漆挥发性成分及其季节性变化。利用水蒸汽法提取了不同季节海漆的挥发性成分，通过GC-MS鉴定其化学组成，发现脂肪酸、二萜和倍半萜是海漆挥发性成分的主要组成，不同季节的挥发性成分差异较大。

Fabrication of Bismuth/Multi-walled Carbon Nanotube Composite Modified Glassy Carbon Electrode for Determination of Cobalt

A bismuth/multi-walled carbon nanotube (Bi/MWNT) composite modified electrode for determination of cobalt by differential pulse adsorptive cathodic stripping voltammetry is described. The electrode is fabricated by potentiostatic pre-plating bismuth film on an MWNT modified glassy carbon (GC) electrode. The Bi/MWNT composite modified electrode exhibits enhanced sensitivity for cobalt detection as compared with the bare GC, MWNT modified and bismuth film electrodes. Numerous key experimental parameters have been examined for optimum analytical performance of the proposed electrode. With an adsorptive accumulation of the Co(II)-dimethylglyoxime complex at -0.8 V for 200 s, the reduction peak current is proportional to the concentration of cobalt in the range of 4.0x10(-11)-1.0x10(-7) mol/L with a lower detection limit of 8.1x10(-11) mol/L. The proposed method has been applied Successfully to cobalt determination in seawater and lake water samples.

Ionophore Based Bismuth Film Electrode for Lead

A Nafion/ionophore, 4-tert-butylcalix[4]arene-tetrakis(N,N-dimethylthioacetamide) composite coated and bismuth film modified glassy carbon electrode. (GC/NA-IONO/BiFE) was described to determine trace lead sensitively and selectively. The characteristics of such modified GC/NA-IONO/BiFE were studied by scanning electron microscopy and cyclic voltammetry. The influence of various experimental parameters upon the stripping lead signal at the GC/NA-IONO/BiFE was explored. Under the optimized conditions, the differential pulse voltammetric stripping response is highly linear over the 0.1-8.0 nM lead range examined (180s preconcentration at -1.2V), with a detection limit of 0.044nM and good precision (RSD=5.4% at 0.5nM). Also applicability to seawater samples was demonstrated at such modified electrode. The high selectivity of ionophore coupled with the excellent electrochemical characteristics of bismuth endow the GC/NA-IONO/BiFE a promising and robust tool for monitoring of trace lead rapidly and precisely.

Carbon isotopic studies of individual lipids in organisms from the Nansha sea area, China

Carbon isotopes of individual lipids in typical organisms from the Nansha sea area were measured by the GC-IRMS analytical technique. delta(13)C values of saturated fatty acids in different organisms examined are from -25.6parts per thousand to -29.7parts per thousand with the average values ranging from -26.4parts per thousand to -28.2parts per thousand and the variance range of 11.8parts per thousand, between different organisms is also observed. Unsaturated fatty acids have heavy carbon isotopic compositions and the mean differences of 2.9%.9-6.8parts per thousand compared to the same carbon number saturated fatty acids. delta(13)C values of n-alkanes range from -27.5%o to -29.7parts per thousand and their mean values, ranging from -28.6parts per thousand, to -28.9parts per thousand, are very close in different organisms. The mean difference in delta(13)C between the saturated fatty acids and n-alkanes is only 1.5parts per thousand, indicating that they have similar biosynthetic pathways. The carbon isotopic variations between the different carbon-number lipids are mostly within +/-2.0parts per thousand, reflecting that they experienced a biosynthetic process of the carbon chain elongation. At the same time, the carbon isotopic genetic relationships between the biological and sedimentary lipids are established by comparative studies of carbon isotopic compositions of individual lipids in organisms and sediments from the Nansha sea area, which provides scientific basis for carbon isotopic applied research of individual lipids.

Concentrations and distribution of organochlorine pesticides in shellfish from Changjiang estuary

To analyze and evaluate the status of organochlorine pollutants in the Changjiang (Yangtze River) estuary and adjacent waters, the concentrations of hexachlorocyclohexane (HCHs) and dichlorodiphenyltrichloroethane (DDTs) in shellfish collected in study area from 2006 to 2007 were determined with gas chromatography (GC). The concentration range of HCHs was (ND-12.13)x10(-3) mg/kg wet weight and averaged at 0.54x10(-3) mg/kg while the concentration of DDTs was in the range of (4.06-281.73) x10(-3) mg/kg with a mean of 57.52x10(-3) mg/kg in the survey areas. The concentrations of DDTs in the shellfish were higher than HCHs', so that DDTs could be considered as typical organochlorine pollutants in the areas. The concentrations of DDTs in the shellfish were higher than HCHs', so that DDTs could be considered as typical organochlorines pollutants. The HCHs in all the shellfish conformed to the first level of criterion (0.02 mg/kg) of the Marion Biology Quality (GB 18421-2001), and that of DDTs in most samples were beyond the first level (0.01 mg/kg) but conformed to the second level (0.10 mg/kg). On average, alpha-HCH and delta-HCH occupied the most part of HCHs, while O,P'-DDT and P,P'-DDT occupied the most part of DDTs. The concentrations of organocholorine pesticides in shellfish samples varied in site and in species. The highest level occurred at the Shengsi (SS), followed by Yangkougang (YKG), Lvsi (LS), Dongyuan (DY) and Beibayao (BBY), low concentrations were observed at Changsha (CS), Beidaodi (BDD), and Gouqi (GQ). The concentration of HCHs and DDTs in most sites decreased clearly from 2006 to 2007 except for YKG, DY, BDD, LYS, and SS. All of above results suggested that the study area was slightly affected by organochlorine pesticide, special by DDTs.

A close phylogenetic relationship between Sipuncula and Annelida evidenced from the complete mitochondrial genome sequence of Phascolosoma esculenta

Background: There are many advantages to the application of complete mitochondrial (mt) genomes in the accurate reconstruction of phylogenetic relationships in Metazoa. Although over one thousand metazoan genomes have been sequenced, the taxonomic sampling is highly biased, left with many phyla without a single representative of complete mitochondrial genome. Sipuncula (peanut worms or star worms) is a small taxon of worm-like marine organisms with an uncertain phylogenetic position. In this report, we present the mitochondrial genome sequence of Phascolosoma esculenta, the first complete mitochondrial genome of the phylum. Results: The mitochondrial genome of P. esculenta is 15,494 bp in length. The coding strand consists of 32.1% A, 21.5% C, 13.0% G, and 33.4% T bases (AT = 65.5%; AT skew = -0.019; GC skew = -0.248). It contains thirteen protein-coding genes (PCGs) with 3,709 codons in total, twenty-two transfer RNA genes, two ribosomal RNA genes and a non-coding AT-rich region (AT = 74.2%). All of the 37 identified genes are transcribed from the same DNA strand. Compared with the typical set of metazoan mt genomes, sipunculid lacks trnR but has an additional trnM. Maximum Likelihood and Bayesian analyses of the protein sequences show that Myzostomida, Sipuncula and Annelida (including echiurans and pogonophorans) form a monophyletic group, which supports a closer relationship between Sipuncula and Annelida than with Mollusca, Brachiopoda, and some other lophotrochozoan groups. Conclusion: This is the first report of a complete mitochondrial genome as a representative within the phylum Sipuncula. It shares many more similar features with the four known annelid and one echiuran mtDNAs. Firstly, sipunculans and annelids share quite similar gene order in the mitochondrial genome, with all 37 genes located on the same strand; secondly, phylogenetic analyses based on the concatenated protein sequences also strongly support the sipunculan + annelid clade (including echiurans and pogonophorans). Hence annelid "key-characters" including segmentation may be more labile than previously assumed.

The complete mitochondrial genome of the ridgetail white prawn Exopalaemon carinicauda Holthuis, 1950 (Crustacean: Decapoda: Palaemonidae) revealed a novel rearrangement of tRNA genes

The complete mitochondrial (mt) DNA sequence was determined for a ridgetail white prawn, Exopalaemon carinicauda Holthuis, 1950 (Crustacea: Decopoda: Palaemonidae). The mt genome is 15,730 bp in length, encoding a standard set of 13 protein-coding genes, 2 ribosomal RNA genes, and 22 transfer RNA genes, which is typical for metazoans. The majority-strand consists of 33.6% A, 23.0% C, 13.4% G, and 30.0% T bases (AT skew = 0.057: GC skew = -0.264). A total of 1045 bp of non-coding nucleotides were observed in 16 intergenic regions,,including a major A+ T rich (79.7%) noncoding region (886 bp). A novel translocation of tRNA(Pro) and tRNA(Thr) was found when comparing this genome with the pancrustacean ground pattern indicating that gene order is not conserved among caridean mitochondria. Furthermore, the rate of Ka/Ks in 13 protein-coding genes between three caridean species is Much less than 1, which indicates a strong Purifying selection within this group. To investigate the phylogenetic relationship within Malacostraca, phylogenetic trees based oil Currently available malacostracan complete mitochondrial sequences were built with the maximum likelihood and Bayesian models. All analyses based oil nucleotide and amino acid data strongly support the monophyly of Decapoda. The Penaeidae, Reptantia, Caridea, and Meiura clades were also recovered as monophyletic groups with Strong Statistical Support. However, the phylogenetic relationships within Pleocyemata are unstable, as represented by the inclusion or exclusion of Caridea. (C) 2009 Elsevier B.V. All rights reserved.

Electrochemical impedance spectroscopy analysis for oxygen reduction reaction in 3.5% NaCl solution

The electrochemical impedance spectroscopy (EIS) at different potentials has been used to study the oxygen reduction reaction (ORR) in 3.5% NaCl solution on glassy carbon (GC) electrode in this work. Results show that ORR consists of three two-electron reaction steps and both superoxide ion (O-2(-)) and hydrogen peroxide (H2O2), which are produced by ORR, obstruct the diffusion of oxygen to the surface of the electrode and make the EIS results change into a transmissive finite diffusion process with the real part contraction and a reflective finite diffusion process from a semi-infinite diffusion process. The values of electron transfer resistance (R-t) and diffusion resistance (R-d) were calculated from EIS. O-2(-) influenced strongly on the Rt values and induced a maximum at -0.45 V.

藏药胡芦巴种籽油脂类成分的GC—MS分析

采用GC-MS法对青海产的胡芦巴油脂类化学成分进行了分析，并以面积归一法测其质量分数。共鉴定出41种化学成分，其中34种为该植物种籽中首次得到。实验中，首先采用三氟化硼-甲醇体系对胡芦巴油进行甲酯化，使酯化更完全，测定效果更好。

高寒灌丛草甸生态系统CO2释放的初步研究

以金露梅(Potentilla fruticosa)灌丛草甸生态系统为对象，应用静态密闭箱-气相色谱法对高寒灌丛(GG)、丛内草甸(GC)和裸地(GL)的CO2释放进行了初步研究。结果表明：GG、GC和GL CO2的释放速率均呈明显的单峰型日变化进程，最大释放速率出现在15：00～17：00之间，最小值在7：00前后出现，白天释放速率大于夜晚；CO2释放速率具有明显的季节性变化特征，生长期CO2释放速率明显高于枯黄期，且均表现为正排放，8月为CO2释放高峰期，释放速率GG>GC>GL(P<0．01)；2003年6月30日至2004年2月28日，高寒灌丛植被-土壤系统CO2释放量为3088．458±287．02g／m^2，丛内草甸植被-土壤系统CO2释放量为2239．685±183．68g／m^2，其中基础土壤呼吸CO2的释放量约为1346．748±176．24g／m^2，分别占GG和GC释放量的43．61％和60．13％；CO2释放速率的日变化主要受地表和5cm地温制约，而季节动态与5cm地温呈显著正相关关系(P<0．01)。

安睡伴侣软胶囊中丁香酚含量测定方法研究

安睡伴侣软胶囊由纯天然传统藏药(包括丁香、肉豆蔻、胡椒、阿魏等6味药材)组方，经超临界CO2萃取其中的挥发油等活性成分后精制而成，具有安神、催眠、抗惊厥等保健功效，无毒副作用。丁香酚是安睡伴侣软胶囊中稳定的、有代表性的功效成分，目前测定药材药品中丁香酚含量的方法主要有液相色谱法(HPLC)和气相色谱法(GC)等。本试验建立了安睡伴侣软胶囊中丁香酚含量的气相色谱和液相色谱测定方法，采用这2种方法测试了5个批次的安睡伴侣软胶囊样品，并将2种方法的测定结果进行了对比。经数据统计分析(配对t检验)显示，HPLC法和GC法所得结果没有显著性差异，2种方法均可满足常规检验要求，都可以作为本制剂的质量检验方法。

海北高寒灌丛草甸生态系统CO2释放速率的季节变化规律

在中国科学院海北高寒草甸生态系统定位站干柴滩地区以金露梅Potentilla fruticosa灌丛草甸生态系统为研究对象,应用静态密闭箱-气相色谱法对高寒灌丛(GG)、丛内草甸(GC)和次生裸地(GL)的CO2释放速率进行了长期观测,并对年释放量作了初步估测.结果表明,GG,GC和GL CO2的释放速率在一年内有明显的季节变化.植物生长季CO2释放量明显高于枯黄期,释放速率GG>GC>GL(P＜0.01),且均表现为正排放.不同季节CO2释放存在明显差异,表现为夏季＞秋季＞春季＞冬季.2003年6月30日至2004年6月28日,高寒灌丛植被-土壤系统CO2释放量为4 293.63±955.75 g/m2,丛内草甸植被-土壤系统CO2释放量为3 319.68±806.19 g/m2,裸地CO2的释放量为1 724.14±444.14 g/m2.CO2释放速率的季节变化与土壤5 cm温度呈显著正相关关系(P＜0.01).