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21.
于2009年3月至2010年1月期间,每月中旬分别从青岛和威海两地区采集人工养殖和野生的栉孔扇贝(Chla-mys farreri)。采集的扇贝经壳长测量后,于闭壳肌血窦中取血,离心,重悬,超声波破碎后制得血细胞破碎液。从抗栉孔扇贝血细胞的单克隆抗体库中筛选出1株与颗粒血细胞和透明血细胞均能结合,并能与血细胞多个蛋白结合的单克隆抗体作为酶联免疫吸附法(ELISA)中的第一抗体;第二抗体为碱性磷酸酶标记的羊抗鼠抗体。将血细胞破碎液包被于酶标板孔中,经一抗、二抗孵育后,显色读数,分析血细胞数量与生长的关系。结果表明:两地区人工养殖和野生扇贝的壳长均由3月的2 cm左右增长到了次年1月的7 cm以上,其中4~7月增长较快,而8~1月相对缓慢。两地区人工养殖和野生扇贝的血细胞数量于3~6月间均维持在较高的水平,6月达到最高峰,随后急剧下降,并于8或9月达到最低值,此后10~1月有所回升,但仍显著低于3~6月的水平。结论认为:栉孔扇贝血细胞数量与生长的关系在两地区、两扇贝品种间差别不大;但3~6月栉孔扇贝生长较快,血细胞数量相对高;而8~10月栉孔扇贝生长缓慢,其血细胞数量较低。 相似文献
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Wesley Bissett Jr. Roger Smith Robert Field Tim Phillips Terry Wade James A. Thompson 《Marine pollution bulletin》2009,58(2):280-286
Locational risks for compromised ecosystem health for the eastern oysters (Crassostrea virginica) harvested from Lavaca Bay, Texas were estimated. Flow cytometric evaluation of variations in DNA content and the lysosomal destabilization assay were used for evaluation of genotoxicity and stress, respectively. Bayesian geo-statistical methods were utilized to estimate and evaluate spatial effects. For models with spatial risks, continuous surface maps of predicted parameter values were created to evaluate risk location. Lysosomal destabilization assay results were spatially oriented whereas flow cytometry results were fit best with the random effects model. While not spatially oriented, the highest levels of variations in DNA content were also present near industrial facilities. Locational risks of increased biomarkers of genotoxicity and stress in the eastern oyster (Crassostrea virginica) were increased with proximity to industrial facilities 相似文献
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Danyang Li Minfang Zheng Yusheng Qiu Limin Lai Nengwang Chen Hongmei Jing Run Zhang Min Chen 《海洋学报(英文版)》2023,42(1):75-82
Nitrogen fixation is one of the most important sources of new nitrogen in the ocean and thus profoundly affects the nitrogen and carbon biogeochemical processes. The distribution, controlling factors, and flux of N2 fixation in the global ocean remain uncertain, partly because of the lack of methodological uniformity. The 15N2 tracer assay (the original bubble method → the 15N2-enriched seawater method → the modified bubble method) is the mainstream method for field measurements of N2 fixation rates (NFRs), among which the original bubble method is the most frequently used. However, accumulating evidence has suggested an underestimation of NFRs when using this method. To improve the availability of previous data, we compared NFRs measured by three 15N2 tracer assays in the South China Sea. Our results indicate that the relationship between NFRs measured by the original bubble method and the 15N2-enriched seawater method varies obviously with area and season, which may be influenced by incubation time, diazotrophic composition, and environmental factors. In comparison, the relationship between NFRs measured by the original bubble method and the modified bubble method is more stable, indicating that the N2 fixation rates based on the original bubble methods may be underestimated by approximately 50%. Based on this result, we revised the flux of N2 fixation in the South China Sea to 40 mmol/(m2·a). Our results improve the availability and comparability of literature NFR data in the South China Sea. The comparison of the 15N2 tracer assay for NFRs measurements on a larger scale is urgently necessary over the global ocean for a more robust understanding of the role of N2 fixation in the marine nitrogen cycle. 相似文献
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用双特异探针技术定性定量分析微型原甲藻 总被引:5,自引:0,他引:5
针对微型原甲藻核糖体大亚基和小亚基RNA分别设计了大亚基探针(LSU probe)和小亚基探针(SSU probe),发展了对微型原甲藻进行定性和定量分析的双特异探针技术.分析数据表明针对微型原甲藻核糖体大亚基RNA的LSU probe能够特异地将微型原甲藻和其他7种微藻分开,而且检测信号的强弱在一定的范围内与细胞数呈线性关系;由于针对微型原甲藻核糖体小亚基的SSU probe探针由于与具齿原甲藻存在核酸序列一致性,该探针与具齿原甲藻有严重的交叉反应,但未发现与海洋原甲藻和其他藻的交叉反应.另外,研究还优化了破碎微型原甲藻细胞所需的超声时间以及获得探针的特异性所需的S1酶反应温度.本研究为实现对微型原甲藻海水样品的快速准确监测奠定了基础. 相似文献
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Biological and procedural factors can influence DNA adduct detection in aquatic organisms. Among them, functional structure and metabolic traits represent major biological determinants for adducts formed by lipophilic pro-mutagenic contaminants. In detecting DNA adducts through the 32P-postlabelling assay, efficiency in DNA purification, digestion, labelling, as well as adduct enrichment and quantification may explain differences between independent studies. Reference DNA adducts have been used to verify some 32P-postlabelling aspects. Data obtained for mussels and fish treated with benzo[a]pyrene (B[a]P) and environmentally exposed to genotoxins confirm the above assertions. Although the 32P-postlabelling assay cannot be proposed for routine biomonitoring it appears a reliable and very sensitive index of exposure to genotoxic pollutants in both fish and mollusks. 相似文献
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The assessment of DNA damage by the Comet assay has been described as a useful non-specific general biomarker of stress in many marine organisms. In field situations it has successfully been employed to distinguish between reference and polluted sites and in the laboratory it has been widely used as a mechanistic tool to determine pollutant effects and mechanisms of DNA damage. To date a wide range of marine vertebrates and invertebrates have been used, however, the usefulness of this assay as a biomarker in cnidarians has not yet been assessed. The aims of this study were to optimize the Comet assay for cnidarian cells and to assess its utility for detecting genotoxic damage in these cells. Cells were isolated from the North American pacific coast temperate sea anemone Anthopleura elegantissima using a non-enzymatic dissociation procedure and viability was determined to be in excess of 90%. Cells were incubated either with (1 h acute exposures) hydrogen peroxide (H(2)O(2)), ethylmethanesulphonate (EMS) or benzo(a)pyrene (B[a]P). In comparison to other marine species, anemone cells exhibited high control or background levels of DNA strand breaks. Despite this, however, we observed dose responses for each of the study chemicals with no reduction in cell viability. This study demonstrates that anemone cells respond to known DNA damaging agents, including B[a]P which requires metabolism to exert its genotoxic effect, and that the Comet assay may prove to be a useful biomarker of stress in cnidarian species. 相似文献