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981.
982.
大鼠颈上神经节烟碱电流的整流及失敏 总被引:1,自引:0,他引:1
在培养的新生大鼠颈上神经节交感神经元标本上,用全细胞膜片钳技术研究了烟碱型乙酰胆碱受体(nAChR)通道的整流及失敏现象。烟碱激动剂引起的全细胞电流在膜电位为负值时随膜电位呈线性改变,而在膜电位达+60mV时仍不出现外向电流,表现出强烈的内向整流。nAChR通道电流存在失敏现象,即持续恒压喷射激动剂所引起的全细胞电流随时间呈指数衰减,不能保持在峰值水平,失敏随激剂浓度呈量效关系,膜电位的超极化也加 相似文献
983.
Ward R. Rice Jeffrey A. Whitsett 《Biochimica et Biophysica Acta (BBA)/Molecular Cell Research》1984,805(3):261-267
Release of [3H]phosphatidylcholine from pulmonary Type II epithelial cells was stimulated by terbutaline, forskolin and cytochalasin D. Compound inhibited both basal and agonist-stimulated release of [3H]PC. The IC50 for inhibition by compound was 1–2 μg/ml, and was similar for inhibition of both basal and stimulated release of [3H]phosphatidylcholine. Inhibitory effects of were noted following a 1 h exposure to compound and persisted up to 3 h. The inhibitory effect of compound was entirely reversed by removing compound from the external milieu. Compound had no effect on cytosolic cyclic AMP levels or lactate dehydrogenase release. Inhibition of surfactant release produced by compound was unaffected by changes in extracellular calcium concentrations. Compound is a non-toxic inhibitor of phosphatidylcholine release from Type II epithelial cells. 相似文献
984.
Wolfgang Schepp Siegfried Ernst Miederer Hans-Jörg Ruoff 《Biochimica et Biophysica Acta (BBA)/Molecular Cell Research》1984,804(2):192-199
Human gastric mucosal cells were isolated from the resected fundic mucosa of peptic ulcer patients. The intracellular content and secretion of intrinsic factor were estimated by binding to cyano[57Co]cobalamin. The content was maximal in the enriched parietal cell fraction which also displayed the highest H+ production as measured by amino[14C]pyrine uptake. Secretagogues evoked full response after 15 min of incubation: pentagastrin (181% of basal secretion), carbachol (208%), histamine (250%) and dibutyryl cyclic adenosine monophosphate (304%). The phosphodiesterase inhibitor isobutylmethylxanthine was slightly more effective even than dibutyryl cAMP. The response to histamine was abolished by ranitidine, indicating activation of adenylate cyclase via histamine H2 receptors, but remained unaffected by atropine, which in turn blocked the carbachol effect, whereas ranitidine was ineffective. The mean formation rate was 8.4 fmol intrinsic factor/106 cells per h under basal conditions and 14.3 fmol in response to histamine. 相似文献
985.
P.J. Thornalley A. Stern 《Biochimica et Biophysica Acta (BBA)/Molecular Cell Research》1984,804(3):308-323
Glyceraldehyde induces changes in the flux of glucose oxidised through the hexose monophosphate pathway, the concentrations of intermediates in the Embden-Meyerhoff pathway, the oxidative status of haemoglobin and levels of reduced and oxidised pyridine nucleotides and glutathione in red cells. Glyceraldehyde autoxidises in the cellular incubations, consuming oxygen and producing glyoxalase I- and II-reactive materials. Major fates of glyceraldehyde in red cells appear to be: (i) adduct formation with reduced glutathione and cellular protein; (ii) autoxidation and reaction with oxyhaemoglobin and pyridine nucleotides, and (iii) phosphorylation of d-glyceraldehyde and entry into the glycolytic pathway as glyceraldehyde 3-phosphate. The production of glycerol from glyceraldehyde by red cell l-hexonate dehydrogenase appears not to be a major reaction of glyceraldehyde in red cells. These results indicate that high concentrations of glyceraldehyde (1–50 mM) may induce oxidative stress in red cells by virtue of the spontaneous autoxidation of glyceraldehyde, forming hydrogen peroxide and α-ketoaldehydes (glyoxalase substrates). The implications of glyceraldehyde-induced oxidative stress for the in vitro anti-sickling effect of dl-glyceraldehyde and for the polyol pathway metabolism of glyceraldehyde are discussed. 相似文献
986.
Neville Marks Martin J. Berg Abba J. Kastin David H. Coy 《Neurochemistry international》1984,6(3):347-353
N-Tyr-MIF-1 (Tyr-Pro-Leu-Gly·NH2), an immunoreactive neuropeptide exhibiting saturable high affinity binding in rat brain was found to be converted into MIF-1 (Pro-Leu-Gly·NH2) by a specific brain aminopeptidase present in rat brain homogenates or cytosol, but with low activity associated with synaptosomal plasma membranes and microsomes. Conversion occurred at a rate of 16 μmol per g w/wt per h and was unaffected by puromycin but inhibited by bestatin (I50, 5 × 10?5 M). Aminopeptidases purified from cytosolic fractions of rat brain (arylamidase), mouse brain (Mn2+-activated aminopeptidase) or porcine kidney (leucine aminopeptidase) were inactive towards N-Tyr-MIF-1 but degraded MIF-1 with release of Leu-Gly·NH2 as detected by RP-HPLC procedures. Morphiceptin (Tyr-Pro-Phe-Pro·NH2), a μ opioid agonist, also acted as a substrate for the N-Tyr-MIF-1 converting enzyme with cleavage of the Tyr-Pro bond. These tetrapeptides, but not MIF-1 or its N-blocked analogs, were degraded in vitro by a metalloendopeptidase purified from kidney membranes. Since dipeptide products were not detected for crude extracts, a significant role for brain metalloendopeptidase on turnover can be excluded. Thus the results point to the presence of a specific (X-Pro-degrading) aminopeptidase in brain cytosol as an enzyme responsible for converting N-Tyr-MIF-1 and inactivating morphiceptin. 相似文献
987.
Massimo De Felici 《Molecular reproduction and development》1984,10(4):423-432
Fluorescent lectins were used to study the chemical nature of carbohydrate moieties present on the surface of female and male germ cells isolated from mouse gonads during fetal and early posnatal development. Concanavalin A (ConA), lens culinaris agglutinin (LCA), ricinus communis agglutinin (RCAI) and wheat germ agglutinin (WGA) bound intensely to the germ cell plasma membrane at all stages studied. Other lectins such as ulex europaeus agglutinin (UEAI) and agglutinin (SBA) did not bind or bound moderately (SBA to female germ cells only). Distinct developmental-related changes were observed when female germ cells were labeled with fluorescein-conjugated peanut agglutinin (PNA) or dolichos biflorus agglutinin (DBA). DBA and PNA binding was absent or weak in fetal female and male germ cells, but became intensely positive in oocytes in the immediate postnatal period. The percentage of oocytes stained with DBA increased during the first three days after birth, and from day 3–4 onwards all oocytes were strongly labeled. I suggest that these changes in lectin binding reflect changes in biochemical structure of the oocyte surface related to differentiative events occurring in the mouse ovary immediately after birth. 相似文献
988.
George E. Milo G. Adolph Ackerman Ronald L. Sanders 《In vitro cellular & developmental biology. Plant》1984,20(12):899-911
Summary Human lung epithelial cells have been isolated and maintained in pure culture and characterized during their time in culture.
Any residual fibroblasts were removed by selective trypsinization within the first 48 h in culture and the residual epithelial
cells from the primary culture grew to confluent density. The epithelial cells at Passage 2 or greater were serially subpassaged
when cultures reached ca. 80% confluency. This procedure permitted us to conduct biochemical and structural studies of starting
materials and subsequent population doublings.
Electron microscope evaluation of both initial monolayers and cell suspensions showed cultures to be composed of a single
cell type. These cells had microvilli on their free or apical surface. Subsequent population doubling level 1 up to 5 exhibited
the same structures. They contained lamellar inclusions, which are typical of Type II alveolar epithelial cells. Fetal lung
(age 18 to 20 wk) cell suspensions processed for electron microscopy before culturing showed cells to be undifferentiated,
epithelial-like with small microvilli along cell borders, and with desmosomes at cell junctions. Lamellar inclusions were
not observed in these cells. Ultrastructural studies of the cultured epithelial cells demonstrated that the lamellar inclusions
had a slightly positive reaction when tested for acid phosphatase. Phospholipid analysis of these lung epithelial cells showed
a phospholipid composition consistent with that found in surfactant-containing Type II cells. Cultured epithelial cells stained
with phosphine 3-R demonstrated a green fluorescent cytoplasm and nucleus with brightly fluorescent yellow-orange perinuclear
particles. The preceding characterization of these cells leads us to conclude that they exhibit structural and biochemical
features commensurate with Type II epithelial cells from human lung. Moreover, these selection techniques applied to the isolation
of human lung Type II cells from the tissue permit us to study the differentiative function of these cells routinely under
conditions of growth in vitro.
This work was supported in part by grants from EPA, R 806638-01 and 131-640-1599A1 相似文献
989.
Shu’a Yagev Michael Heller Arié Pinson 《In vitro cellular & developmental biology. Plant》1984,20(12):893-898
Summary Postnatal rat heart cells in culture enriched with respect to muscle cells were obtained by either high density seeding or
by the replating technique. [3H]Thymidine incorporation to DNA and the enzymatic pattern of cytoplasmic and lysosomal enzymes have been studied as a function
of the culture’s age, of seeding density, and replating. It was shown that (a) replating maintains predominance of myocyte
population for at least 2 wk in culture; (b) heavy seeding density allows homogeneous myocyte population for the 1st wk in
culture; and (c) the enzyme profile of the culture may serve as an indicator for the type of cell population in culture and
its state of differentiation.
This study was done as partial fulfilment of the M.Sc. thesis in Biochemistry (SY). Supported by grants from The Chief Scientist,
Ministry of Health, State of Israel; The Ministry of Education and Sciences, State of Niedersachssen (FRG); and The Foundation
for Heart Research from Mr. and Mrs. D. Vidal-Madjar, Paris, France. 相似文献
990.
K. Norrby S. Bergström P. Druvefors 《In vitro cellular & developmental biology. Plant》1984,20(8):607-614
Summary The intact membranous rat mesentery was cultured in Eagle's minimum essential medium containing no serum or only low concentrations
of serum. The procedure is in some important respects superior to previous organ culture techniques. To estimate the extent
of disturbance of homeostasis of the tissue in culture, the spontaneous mast-cell histamine release was quantitated after
preculture preparation of the specimens and after different intervals in culture. Also, the proliferation of fibroblasts and
mesothelial cells that predominate in the mesentery was assessed at 48 h by cytofluorometric quantitation of DNA in single-tissue
cells.
Spontaneous histamine release was time dependent during cultivation, amounting to ca. 50% at 48 h, and was affected by the
medium used for moistening the tissue before cultivation. Culturing also brought about great spontaneous increase in the proliferation
of fibroblasts and mesothelial cells, the rate being related to the concentration of serum. Addition of the mast-cell secretagogues
48/80 or polymyxin B at 1 h caused rapid release of 50 to 60% of the histamine and was followed by augmented proliferation
in the serum-containing media.
The spontaneous increase of cell proliferation in tissue culture may be causally related to mast-cell secretion. Further studies
are needed to define factors influencing the spontaneous mast-cell secretion and the mast-cell-dependent mitogenesis in normal
tissue cells
Supported by grants from the Swedish Medical Research Council (Project 5942) and State Board for Animal Experiments. 相似文献