全文获取类型
收费全文 | 6950篇 |
免费 | 576篇 |
国内免费 | 1篇 |
专业分类
7527篇 |
出版年
2023年 | 33篇 |
2022年 | 83篇 |
2021年 | 193篇 |
2020年 | 111篇 |
2019年 | 136篇 |
2018年 | 193篇 |
2017年 | 155篇 |
2016年 | 208篇 |
2015年 | 318篇 |
2014年 | 378篇 |
2013年 | 459篇 |
2012年 | 533篇 |
2011年 | 575篇 |
2010年 | 330篇 |
2009年 | 315篇 |
2008年 | 387篇 |
2007年 | 416篇 |
2006年 | 369篇 |
2005年 | 320篇 |
2004年 | 309篇 |
2003年 | 292篇 |
2002年 | 257篇 |
2001年 | 100篇 |
2000年 | 78篇 |
1999年 | 87篇 |
1998年 | 66篇 |
1997年 | 49篇 |
1996年 | 47篇 |
1995年 | 58篇 |
1994年 | 42篇 |
1993年 | 44篇 |
1992年 | 57篇 |
1991年 | 32篇 |
1990年 | 55篇 |
1989年 | 50篇 |
1988年 | 41篇 |
1987年 | 25篇 |
1986年 | 26篇 |
1985年 | 36篇 |
1984年 | 33篇 |
1983年 | 24篇 |
1982年 | 21篇 |
1981年 | 20篇 |
1980年 | 12篇 |
1979年 | 19篇 |
1978年 | 17篇 |
1977年 | 21篇 |
1976年 | 15篇 |
1973年 | 11篇 |
1972年 | 11篇 |
排序方式: 共有7527条查询结果,搜索用时 0 毫秒
111.
Controlling accumulations of unwanted biofilms requires an understanding of the mechanisms that organisms use to interact with submerged substrata. While the substratum properties influencing biofilm formation are well studied, those that may lead to cellular or biofilm detachment are not. Surface-grafted stimuli-responsive polymers, such as poly (N-isopropylacrylamide) (PNIPAAm) release attached cells upon induction of environmentally-triggered phase changes. Altering the physicochemical characteristics of such polymeric systems for systematically studying release, however, can alter the phase transition. The physico-chemical changes of thin films of PNIPAAm grafted from initiator-modified self-assembled monolayers (SAMs) of ω-substituted alkanethiolates on gold can be altered by changing the composition of the underlying SAM, without affecting the overlying polymer. This work demonstrates that the ability to tune such changes in substratum physico-chemistry allows systematic study of attachment and release of bacteria over a large range of water contact angles. Such surfaces show great promise for studying a variety of interactions at the biointerface. Understanding of the source of this tunability will require further studies into the heterogeneity of such films and further investigation of interactions beyond those of water wettability. 相似文献
112.
Epipelagic siphonophore assemblages associated with water masses along a transect between Chile and Easter Island (eastern South Pacific Ocean) 总被引:1,自引:0,他引:1
We analyze for the first time the spatial distribution of siphonophoresin relation to water masses along a 3750-km oceanic transectbetween the Chilean coast and the Easter Island (27° S),a sector scarcely known of eastern South Pacific Ocean. Thirty-onesiphonophore species were identified; Sulculeolaria turgidaand Vogtia glabra were recorded for the first time in this sector.The most abundant species were Muggiaea atlantica (29.2%), Eudoxoidesspiralis (24.5%) and Lensia subtilis (13.1%). Two differentsiphonophore assemblages east and west of 76 W, associated respectivelywith Subantarctic Water and Subtropical Water masses, can beused as water mass indicators. The former included the threemost abundant species, whereas the latter showed greater speciesrichness. This study provides basic knowledge on spatial distributionof siphonophores, which is important to develop future researchfocused on understanding the ecological role and biologicalprocesses driven by planktonic organisms in the southeasternPacific Ocean. 相似文献
113.
Pseudo‐nitzschia arctica sp. nov., a new cold‐water cryptic Pseudo‐nitzschia species within the P. pseudodelicatissima complex
下载免费PDF全文
![点击此处可从《Journal of phycology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
114.
Falvo S Di Carli M Desiderio A Benvenuto E Moglia A America T Lanteri S Acquadro A 《Proteomics》2012,12(3):448-460
Plants respond to ultraviolet stress inducing a self-defence through the regulation of specific gene family members. The UV acclimation is the result of biochemical and physiological processes, such as enhancement of the antioxidant enzymatic system and accumulation of UV-absorbing phenolic compounds (e.g. flavonoids). Globe artichoke is an attractive species for studying the protein network involved in UV stress response, being characterized by remarkable levels of inducible antioxidants. Proteomic tools can assist the evaluation of the expression patterns of UV-responsive proteins and we applied the difference in-gel electrophoresis (DIGE) technology for monitoring the globe artichoke proteome variation at four time points following an acute UV-C exposure. A total of 145 UV-C-modulated proteins were observed and 119 were identified by LC-MS/MS using a ~144,000 customized Compositae protein database, which included about 19,000 globe artichoke unigenes. Proteins were Gene Ontology (GO) categorized, visualized on their pathways and their behaviour was discussed. A predicted protein interaction network was produced and highly connected hub-like proteins were highlighted. Most of the proteins differentially modulated were chloroplast located, involved in photosynthesis, sugar metabolisms, protein folding and abiotic stress. The identification of UV-C-responsive proteins may contribute to shed light on the molecular mechanisms underlying plant responses to UV stress. 相似文献
115.
Optimized construction of low-redundancy cDNA mini-libraries using low-stringency RT-PCR is described cDNAs are generated using arbitrary consensus-degenerate hybrid oligonucleotide primers and nanogram amounts of Schistosoma mansoni mRNA. A number of conditions such as temperature and salt concentration are combined to create balanced, low-stringency conditions that permit a normalized amplification of a diversified, random set of sequences. On average, 350 different sequences are obtained per mini-library, which represents a significantly higher diversity of messages per library when compared to previously published conditions (ie., 20-40 sequences/ mini-library). The optimized high-throughput approach described here is likely to help in the discovery of expressed genes in any complex organism. 相似文献
116.
Nonphosphorylating glyceraldehyde-3-phosphate dehydrogenase (np-Ga3PDHase) is a cytosolic unconventional glycolytic enzyme of plant cells regulated by phosphorylation in heterotrophic tissues. After interaction with 14-3-3 proteins, the phosphorylated enzyme becomes less active and more sensitive to regulation by adenylates and inorganic pyrophosphate. Here, we acknowledge that in wheat (Triticum aestivum), np-Ga3PDHase is specifically phosphorylated by the SnRK (SNF1-related) protein kinase family. Interestingly, only the kinase present in heterotrophic tissues (endosperm and shoots, but not in leaves) was found active. The specific SnRK partially purified from endosperm exhibited a requirement for Mg(2+) or Mn(2+) (being Ca(2+) independent), having a molecular mass of approximately 200 kD. The kinase also phosphorylated standard peptides SAMS, AMARA, and SP46, as well as endogenous sucrose synthase, results suggesting that it could be a member of the SnRK1 subfamily. Concurrently, the partially purified wheat SnRK was recognized by antibodies raised against a peptide conserved between SnRK1s from sorghum (Sorghum bicolor) and maize (Zea mays) developing seeds. The wheat kinase was allosterically inhibited by ribose-5-phosphate and, to a lesser extent, by fructose-1,6-bisphosphate and 3-phosphoglycerate, while glucose-6-phosphate (the main effector of spinach [Spinacia oleracea] leaves, SnRK1) and trehalose-6-phosphate produced little or no effect. Results support a distinctive allosteric regulation of SnRK1 present in photosynthetic or heterotrophic plant tissues. After in silico analysis, we constructed two np-Ga3PDHase mutants, S404A and S447A, identifying serine-404 as the target of phosphorylation. Results suggest that both np-Ga3PDHase and the specific kinase could be under control, critically affecting the metabolic scenario involving carbohydrates and reducing power partition and storage in heterotrophic plant cells. 相似文献
117.
A partially purified D-xylose isomerase has been isolated from cells of Streptomyces albus NRRL 5778 and some of its properties have been determined. D-Glucose, D-xylose, D-ribose, L-arabinose, and L-rhamnose served as substrates for the enzyme with respective Km values of 86, 93, 350, 153, and 312 mM and Vmax values measuring 1.23, 2.9, 2.63, 0.153, and 0.048 mumol min per mg of protein. The hexose D-allose was also isomerized. The enzyme was strongly activated by 1.0 mM Mg2+ but only partially activated by 1.0 mM Co2+. The respective Km values for Mg2+ and Co2+ were 0.3 and 0.003 mM. Mg2+ and Co2+ appear to have separate binding sites on the isomerase. These cations also protect the enzyme from thermal denaturation and from D-sorbitol inhibition. The optimum temperature for ketose formation was 70 to 80 C at pH values ranging from 7 to 9. D-Sorbitol acts as a competitive inhibitor with a Ki of 5.5 mM against D-glucose, D-xylose, and D-ribose. Induction experiments, Mg2+ activation, and D-sorbitol inhibition indicated that a single enzyme (D-xylose isomerase) was responsible for the isomerization of the pentoses, methyl pentose, and glucose. 相似文献
118.
Bienvenut WV Déon C Pasquarello C Campbell JM Sanchez JC Vestal ML Hochstrasser DF 《Proteomics》2002,2(7):868-876
Although peptide mass fingerprinting is currently the method of choice to identify proteins, the number of proteins available in databases is increasing constantly, and hence, the advantage of having sequence data on a selected peptide, in order to increase the effectiveness of database searching, is more crucial. Until recently, the ability to identify proteins based on the peptide sequence was essentially limited to the use of electrospray ionization tandem mass spectrometry (MS) methods. The recent development of new instruments with matrix-assisted laser desorption/ionization (MALDI) sources and true tandem mass spectrometry (MS/MS) capabilities creates the capacity to obtain high quality tandem mass spectra of peptides. In this work, using the new high resolution tandem time of flight MALDI-(TOF/TOF) mass spectrometer from Applied Biosystems, examples of successful identification and characterization of bovine heart proteins (SWISS-PROT entries: P02192, Q9XSC6, P13620) separated by two-dimensional electrophoresis and blotted onto polyvinylidene difluoride membrane are described. Tryptic protein digests were analyzed by MALDI-TOF to identify peptide masses afterward used for MS/MS. Subsequent high energy MALDI-TOF/TOF collision-induced dissociation spectra were recorded on selected ions. All data, both MS and MS/MS, were recorded on the same instrument. Tandem mass spectra were submitted to database searching using MS-Tag or were manually de novo sequenced. An interesting modification of a tryptophan residue, a "double oxidation", came to light during these analyses. 相似文献
119.
Covarrubias SA de-Bashan LE Moreno M Bashan Y 《Applied microbiology and biotechnology》2012,93(6):2669-2680
When the freshwater microalga Chlorella sorokiniana and the plant growth-promoting bacterium Azospirillum brasilense were deployed as free suspensions in unsterile, municipal wastewater for tertiary wastewater treatment, their population
was significantly lower compared with their populations in sterile wastewater. At the same time, the numbers of natural microfauna
and wastewater bacteria increased. Immobilization of C. sorokiniana and A. brasilense in small (2–4 mm in diameter), polymer Ca-alginate beads significantly enhanced their populations when these beads were suspended
in normal wastewater. All microbial populations within and on the surface of the beads were evaluated by quantitative fluorescence
in situ hybridization combined with scanning electron microscopy and direct measurements. Submerging immobilizing beads in
wastewater created the following sequence of events: (a) a biofilm composed of wastewater bacteria and A. brasilense was created on the surface of the beads, (b) the bead inhibited penetration of outside organisms into the beads, (c) the
bead inhibited liberation of the immobilized microorganisms into the wastewater, and (d) permitted an uninterrupted reduction
of ammonium and phosphorus from the wastewater. This study demonstrated that wastewater microbial populations are responsible
for decreasing populations of biological agents used for wastewater treatment and immobilization in alginate beads provided
a protective environment for these agents to carry out uninterrupted tertiary wastewater treatment. 相似文献
120.
Comparative analysis reveals amino acids critical for anticancer activity of peptide CIGB‐552
下载免费PDF全文
![点击此处可从《Journal of peptide science》网站下载免费的PDF全文](/ch/ext_images/free.gif)
Soledad Astrada Yolanda Gomez Exequiel Barrera Gonzalo Obal Otto Pritsch Sergio Pantano Maribel G. Vallespí Mariela Bollati‐Fogolín 《Journal of peptide science》2016,22(11-12):711-722
Because of resistance development by cancer cells against current anticancer drugs, there is a considerable interest in developing novel antitumor agents. We have previously demonstrated that CIGB‐552, a novel cell‐penetrating synthetic peptide, was effective in reducing tumor size and increasing lifespan in tumor‐bearing mice. Studies of protein–peptide interactions have shown that COMMD1 protein is a major mediator of CIGB‐552 antitumor activity. Furthermore, a typical serine‐protease degradation pattern for CIGB‐552 in BALB/c mice serum was identified, yielding peptides which differ from CIGB‐552 in size and physical properties. In the present study, we show the results obtained from a comparative analysis between CIGB‐552 and its main metabolites regarding physicochemical properties, cellular internalization, and their capability to elicit apoptosis in MCF‐7 cells. None of the analyzed metabolites proved to be as effective as CIGB‐552 in promoting apoptosis in MCF‐7. Taking into account these results, it seemed important to examine their cell‐penetrating capacity and interaction with COMMD1. We show that internalization, a lipid binding‐dependent process, is impaired as well as metabolite–COMMD1 interaction, key component of the apoptotic mechanism. Altogether, our results suggest that features conferred by the amino acid sequence are decisive for CIGB‐552 biological activity, turning it into the minimal functional unit. Copyright © 2016 European Peptide Society and John Wiley & Sons, Ltd. 相似文献