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61.
Horacio Gómez-Acevedo Michael Y. Li Steven Jacobson 《Bulletin of mathematical biology》2010,72(3):681-696
Human T-cell leukaemia/lymphoma virus type I (HTLV-I) is a retrovirus that has been identified as the causative agent of HTLV-I-associated
myelopathy/tropical spastic paraparesis (HAM/TSP) and other illnesses. HTLV-I infects primarily CD4+ T cells and the transmission occurs through direct cell-to-cell contact. HAM/TSP patients harbor higher proviral loads in
peripheral blood lymphocytes than asymptomatic carriers. Also, HAM/TSP patients exhibit a remarkably high number of circulating
HTLV-I-specific CD8+ cytotoxic T lymphocytes (CTLs) in the peripheral blood. While CTLs have a protective role by killing the infected cells and
lowering the proviral load, a high level of CTLs and their cytotoxicity are believed to be a main cause of the development
of HAM/TSP. A mathematical model for HTLV-I infection of CD4+ T cells that incorporates the CD8+ cytotoxic T-cell (CTL) response is investigated. Our mathematical analysis reveals that the system can stabilize at a carrier
steady-state with persistent viral infection but no CTL response, or at a HAM/TSP steady-state at which both the viral infection
and CTL response are persistent. We also establish two threshold parameters R
0 and R
1, the basic reproduction numbers for viral persistence and for CTL response, respectively. We show that the parameter R
1 can be used to distinguish asymptomatic carriers from HAM/TSP patients, and as an important control parameter for preventing
the development of HAM/TSP. 相似文献
62.
Jennifer L. Belsky Alyssa J. Ashley Premal A. Bhatt Karen V. Gilbert Heather R. Joyce Chunhua Pan Horacio Pappa Samir Z. Wahab 《AAPS PharmSciTech》2010,11(2):994-1004
The water-insoluble procedures in US Pharmacopeia (USP) General Chapter Residual Solvents <467>, which are based on European Pharmacopoeia procedures, were optimized and modified before their inclusion in the chapter to improve their scope, performance, and ruggedness.
The optimized procedures use a static headspace introduction system with a gas chromatograph equipped with a flame ionization
detector. This article describes some of the key changes made to the USP published procedures, including use of dimethyl sulfoxide
(DMSO) or dimethylformamide (DMF) as the solvent, addition of 5 mL of water and 1 mL of sample (dissolved in DMSO or DMF)
to the headspace vial, use of a 3:1 GC split ratio, and use of new matrix-matched system suitability solutions. These procedures
were verified with two different active pharmaceutical ingredients—hydroxyzine pamoate and prednisone. In the investigation,
the more polar material (hydroxyzine pamoate) showed greater recoveries for the optimized procedures when prepared in DMSO.
The less polar material (prednisone) typically had greater recoveries in DMF for the optimized procedures. During experimentation,
insights into sample preparation, additional types of headspace instrumentation, solvent purity, and other parameters were
also gained. 相似文献
63.
Varela Eduardo Sousa; Beasley Colin Robert; Schneider Horacio; Sampaio Iracilda; Marques-Silva Nelane Do Socorro; Tagliaro Claudia Helena 《Journal of Molluscan Studies》2007,73(3):229-234
As a result of phenotypic plasticity, the cupped oysters (Crassostrea)are difficult to identify by means of their morphology. However,molecular DNA markers are a useful means of discriminating amongthese species. Cupped oysters are one of the most widely culturedmarine invertebrates and correct species identification is importantin aquaculture. Moreover, the molecular phylogeny of the genusCrassostrea and the subfamily Crassostreinae is still not clear.In order to identify the Brazilian cupped oysters and to clarifythe phylogenetic relationships of these species, we sequenceda fragment of mitochondrial DNA (16S rRNA gene) from 120 specimenscollected at nine different sites distributed along the Braziliancoast. The results identified two native species of oyster:Crassostrea gasar, from the Amazon to the Parnaíba delta;and Crassostrea rhizophorae, from the northeast (Fortim) tothe south of Brazil. An exotic Crassostrea species, closelyrelated to Indo-Pacific Crassostrea, was found in one locationin the north of Brazil. Crassostrea showed monophyly and theAtlantic oysters are clearly separated from the Indo-Pacificcluster. (Received 30 May 2006; accepted 12 April 2007) 相似文献
64.
Grigorios A. Papadakos Horacio Nastri Paul Riggs Cynthia M. Dupureur 《Journal of biological inorganic chemistry》2007,12(4):557-569
The hydrolysis of phosphodiester bonds by nucleases is critical to nucleic acid processing. Many nucleases utilize metal ion cofactors, and for a number of these enzymes two active-site metal ions have been detected. Testing proposed mechanistic roles for individual bound metal ions has been hampered by the similarity between the sites and cooperative behavior. In the homodimeric PvuII restriction endonuclease, the metal ion dependence of DNA binding is sigmoidal and consistent with two classes of coupled metal ion binding sites. We reasoned that a conservative active-site mutation would perturb the ligand field sufficiently to observe the titration of individual metal ion binding sites without significantly disturbing enzyme function. Indeed, mutation of a Tyr residue 5.5 A from both metal ions in the enzyme-substrate crystal structure (Y94F) renders the metal ion dependence of DNA binding biphasic: two classes of metal ion binding sites become distinct in the presence of DNA. The perturbation in metal ion coordination is supported by 1H-15N heteronuclear single quantum coherence spectra of enzyme-Ca(II) and enzyme-Ca(II)-DNA complexes. Metal ion binding by free Y94F is basically unperturbed: through multiple experiments with different metal ions, the data are consistent with two alkaline earth metal ion binding sites per subunit of low millimolar affinity, behavior which is very similar to that of the wild type. The results presented here indicate a role for the hydroxyl group of Tyr94 in the coupling of metal ion binding sites in the presence of DNA. Its removal causes the affinities for the two metal ion binding sites to be resolved in the presence of substrate. Such tuning of metal ion affinities will be invaluable to efforts to ascertain the contributions of individual bound metal ions to metallonuclease function. 相似文献
65.
Heras H Dreon MS Ituarte S Pollero RJ 《Comparative biochemistry and physiology. Toxicology & pharmacology : CBP》2007,146(1-2):158-167
Carotenoid-binding proteins are commonly found in invertebrates. Their carotenoids form non-covalent complexes with proteins giving tissues a variety of colors. In molluscs they have been described in only a few species. In particular, the egg perivitellin fluid of those Ampullariid species which deposit eggs above the waterline is provided with carotenoproteins playing several roles ranging from photoprotection, antioxidant or antitrypsin actions to nutrient provision for development. These molecules form complex glyco-lipo-carotenoproteins of high molecular weight where either free astaxanthin (3,3'-dihydroxy-beta, beta'-carotene- 4,4'dione) or astaxanthin esterified with fatty acids, occur more frequently. This review compiles the current knowledge on the biochemical composition and biophysical data on the chemical and thermal stability of egg carotenoproteins in ampullariid. In addition, recent data on their metabolism, their cellular site of biosynthesis during perivitellogenesis, as well as their carotenoid binding properties are reviewed, highlighting the physiological significance of carotenoproteins in the context of the reproductive biology of these molluscs. 相似文献
66.
Melissa Richard-Greenblatt Horacio Bach John Adamson Sandra Pe?a-Diaz Wu Li Adrie J. C. Steyn Yossef Av-Gay 《The Journal of biological chemistry》2015,290(38):23064-23076
Ergothioneine (EGT) is synthesized in mycobacteria, but limited knowledge exists regarding its synthesis, physiological role, and regulation. We have identified Rv3701c from Mycobacterium tuberculosis to encode for EgtD, a required histidine methyltransferase that catalyzes first biosynthesis step in EGT biosynthesis. EgtD was found to be phosphorylated by the serine/threonine protein kinase PknD. PknD phosphorylates EgtD both in vitro and in a cell-based system on Thr213. The phosphomimetic (T213E) but not the phosphoablative (T213A) mutant of EgtD failed to restore EGT synthesis in a ΔegtD mutant. The findings together with observed elevated levels of EGT in a pknD transposon mutant during in vitro growth suggests that EgtD phosphorylation by PknD negatively regulates EGT biosynthesis. We further showed that EGT is required in a nutrient-starved model of persistence and is needed for long term infection of murine macrophages. 相似文献
67.
Borcard F Godinat A Staedler D Blanco HC Dumont AL Chapuis-Bernasconi C Scaletta C Applegate LA Juillerat FK Gonzenbach UT Gerber-Lemaire S Juillerat-Jeanneret L 《Bioconjugate chemistry》2011,22(7):1422-1432
The chemical functionalization of cell-surface proteins of human primary fetal bone cells with hydrophilic bioorthogonal intermediates was investigated. Toward this goal, chemical pathways were developed for click reaction-mediated coupling of alkyne derivatives with cellular azido-expressing proteins. The incorporation via a tetraethylene glycol linker of a dipeptide and a reporter biotin allowed the proof of concept for the introduction of cell-specific peptide ligands and allowed us to follow the reaction in living cells. Tuning the conditions of the click reaction resulted in chemical functionalization of living human fetal osteoblasts with excellent cell survival. 相似文献
68.
de las Heras A Cain RJ Bielecka MK Vázquez-Boland JA 《Current opinion in microbiology》2011,14(2):118-127
Listeria monocytogenes is the causative agent of listeriosis, a severe foodborne infection. These bacteria live as soil saprotrophs on decaying plant matter but also as intracellular parasites, using the cell cytosol as a replication niche. PrfA, a regulatory protein, integrates a number of environmental cues that signal the transition between these two contrasting lifestyles, activating a set of key virulence factors during host infection. While a number of details concerning the general mode of action of this virulence master switch have been elucidated, others remain unsolved. Recent work has revealed additional mechanisms that contribute to L. monocytogenes virulence modulation, often via cross-talk with PrfA, or by regulating new genes involved in host colonization. 相似文献
69.
Chicken egg yolk antibodies (IgY-technology): a review of progress in production and use in research and human and veterinary medicine 总被引:10,自引:0,他引:10
Schade R Calzado EG Sarmiento R Chacana PA Porankiewicz-Asplund J Terzolo HR 《Alternatives to laboratory animals : ATLA》2005,33(2):129-154
The production of antibodies (Abs) in chickens and the extraction of specific Abs from egg yolk (IgY Abs) are increasingly attracting the interest of the scientific community, as demonstrated by the significant growth of the IgY literature. This review offers detailed and comprehensive information about IgY-technology, including: a) possibilities for hen keeping in accordance with the Three Rs principles; b) new insights into the IgY transfer mechanism from blood to yolk as a biological basis for the technology; c) the comparative characteristics of IgY Abs and IgG Abs; d) the high efficacy of the technique, in view of the extraordinary amount of IgY Ab produced by one hen in one year (between 20 g and 40 g IgY in total); e) comparisons between the efficacies of IgY Abs and IgG Abs (rabbit, sheep, mouse) in several immunological assays; f) immunisation protocols, as well as the most commonly used IgY-extraction procedures; g) new possibilities for application in human and veterinary medicine, including strategies for the treatment of Helicobacter pylori infection or fatal intestinal diseases in children, particularly in poor countries, for reducing the use of antibiotics, and, in Asia and South America, for producing Abs against snake, spider and scorpion venoms; and h) the use of IgY Abs in various fields of research, also taking into consideration recent developments in South America (particularly Argentina and Cuba) and in Asia. 相似文献
70.
Roles of the Ras-MEK-mitogen-activated protein kinase and phosphatidylinositol 3-kinase-Akt-mTOR pathways in Jaagsiekte sheep retrovirus-induced transformation of rodent fibroblast and epithelial cell lines 总被引:2,自引:0,他引:2 下载免费PDF全文
Jaagsiekte sheep retrovirus (JSRV) is the causative agent of ovine pulmonary adenocarcinoma (OPA), a transmissible lung cancer of sheep. The virus can induce tumors rapidly, and we previously found that the JSRV envelope protein (Env) functions as an oncogene, because it can transform mammalian and avian fibroblast cell lines. (N. Maeda, Proc. Natl. Acad. Sci. USA 98:4449-4454, 2001). The molecular mechanisms of JSRV Env transformation are of considerable interest. Several reports suggested that the phosphatidylinositol 3-kinase/Akt pathway is important for transformation of mammalian fibroblasts but not for chicken fibroblasts. In this study, we found that Akt/mTOR is involved in JSRV transformation of mouse NIH 3T3 fibroblasts, because treatment with the mTOR inhibitor rapamycin reduced transformation. We also found that H/N-Ras inhibitor FTI-277 and MEK1/2 inhibitors PD98059 and U0126 strongly inhibited JSRV transformation of NIH 3T3 fibroblasts, suggesting that the H/N-Ras-MEK-mitogen-activated protein kinase (MAPK) p44/42 pathway is necessary for the transformation. In RK3E epithelial cells, the MEK1/2 inhibitors also eliminated transformation, but FTI-277 only partially inhibited transformation. It was noteworthy that p38 MAPK inhibitors enhanced JSRV transformation in both fibroblasts and epithelial cells. Treatment of transformed cells with p38 inhibitors both increased levels of phospho-MEK1/2 and phospho-p44/42 and induced rapid enhancement of the transformed phenotype. Immunohistochemical staining of tumor tissues from naturally and experimentally induced OPA and naturally occurring enzootic nasal adenocarcinoma revealed strong activation of MAPK p44/42 in all cases examined. However, p38 activation was not generally observed. These results indicate that signaling through two pathways (in particular, H/N-Ras-MEK-MAPK and, to a lesser extent, Akt-mTOR) is important for JSRV-induced transformation and that p38 MAPK has a negative regulatory effect on transformation, perhaps via MEK1/2 and p44/42. 相似文献