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101.
Chemical modification of Rhodospirillum rubrum chromatophores by 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole (NBD-Cl) results in inactivation of photophosphorylation, Mg2+-ATPase, oxidative phosphorylation and ATP-driven transhydrogenase, with apparent first-order kinetics. Other energy-linked reactions such as light-driven transhydrogenase and light-dependent proton uptake were insensitive to NBD-Cl. The Ca2+-ATPase activity of the soluble coupling factor from chromatophores (R. rubrum F1) was inactivated by NBD-Cl with kinetics resembling those described for Mg2+-ATPase and photophosphorylation activities of chromatophores. Both NBD-chromatophores and NBD-R. rubrum F1 fully recovered their activities when subjected to thiolysis by dithioerythritol. Phosphoryl transfer reactions of chromatophores and Ca2+-ATPase activity of R. rubrum F1 were fully protected by 5 mM Pi against modification by NBD-Cl. ADP or ATP afforded partial protection. Analysis of the protection of Ca2+-ATPase activity by Pi indicated that NBD-Cl and Pi are mutually exclusive ligands. Spectroscopic studies revealed that tyrosine and sulfhydryl residues in R. rubrum F1 underwent modification by NBD-Cl. However, the inactivation was only related to the modification of tyrosine groups. 相似文献
102.
Summary An electron microscopic investigation of the extrapulmonary respiratory tract of embryos and chick of the domestic fowl (Gallus domesticus) has demonstrated for the first time in birds the presence here of a small number of epithelial cells characterised by an aminecontaining type of granule. These granular cells were scattered singly throughout the trachea, syrinx and primary bronchi and seemed more numerous in the caudal part of the airway. In favourable planes of section a small part of the cell was in contact with the luminal surface of the epithelium. The characteristic granular vesicles (approximate diameter 140 nm) appeared to be randomly distributed in the cytoplasm and there was no concentration of vesicles close to the plasma membrane. One of the cells was closely associated with an intraepithelial axon. By fluorescence microscopy, a small number of cells with a similar shape and distribution to the granular cells was observed after administration of 3,4-dihydroxyphenylalanine which may indicate the presence of an amine handlign mechanism in these cells. It is suggested that the granular cells belong to the APUD system of endocrine cells and that they may be modulated by the concentration of gas in the airways. 相似文献
103.
Activation of phosphorylating vesicles by net transfer of phosphatidyl choline by phospholipid transfer protein 总被引:2,自引:0,他引:2
Y Kagawa L W Johnson E Racker 《Biochemical and biophysical research communications》1973,50(2):245-251
Vesicles formed with phosphatidyl ethanolamine, phosphatidyl choline, cardiolipin, coupling factors and hydrophobic proteins from bovine heart mitochondria catalyzed a rapid32Pi-ATP exchange. When phosphatidyl choline was deleted during the assembly of the vesicles, little32Pi-ATP exchange was observed. Exchange activity was induced by incubating such deficient vesicles with phosphatidyl choline liposomes in the presence of a phosphatidyl choline transfer protein isolated from bovine heart. Transfer of [32P] phosphatidyl choline was demonstrated by isolation of the activated vesicles by sucrose density centrifugation. 相似文献
104.
105.
R. Joplin A. J. Strain J. M. Neuberger 《In vitro cellular & developmental biology. Plant》1989,25(12):1189-1192
Summary Biliary epithelial cells (BEC) lining the intra-hepatic biliary ducts are the site of damage in several immunologically mediated
liver diseases. BEC are difficult to isolate since they represent only 5% of the total cell number in normal liver. In this
communication, a novel method for their isolation from normal liver is presented using a monoclonal antibody (HEA125) with
specificity for an epithelial cell surface glyco-protein reported to be expressed in liver only by biliary epithelium. By
combining differential density centrifugation and immuno-magnetic separation using HEA125 pure BEC (105 cells/g fresh tissue) were prepared routinely. These cells were maintained in culture for up to 4 weeks with significant
increases in cell numbers. The ability to prepare BEC from human liver offers an opportunity to develop In Vitro models to
investigate the aetiology of diseases in intra-hepatic biliary epithelium.
EDITOR’S STATEMENT This is a novel application to purification of specific liver cell types directly from tissue. It is well-suited
for rapid communication because of its novelty and potential utility to investigators. 相似文献
106.
107.
James C. Beck Howard L. Hosick Bruce A. Watkins 《In vitro cellular & developmental biology. Plant》1989,25(5):409-418
Summary We investigated the effects of conditioned media derived from mouse mammary fat pads on the proliferation of CL-S1 cells,
an epithelial cell line originally isolated from a preneoplastic mammary outgrowth line. Cell proliferation in vitro in serum-free
defined medium was compared to that in this medium conditioned using intact mammary fat pad pieces or isolated fat pad adipocytes.
Culture medium was conditioned by incubating the conditioning material in defined culture medium for 24 h at 37°C. Conditioned
medium induced CL-S1 proliferation as much as 10- to 20-fold above the minimal levels of growth in control cultures after
13 d of culture. The growth-stimulatory factor(s) had an apparent molecular weight of greater than 10 kDa. This growth-stimulatory
activity was both heat and trypsin stable. Because the role of adipose tissue is to store and release lipids, we next tested
whether lipids are released during medium conditioning. The lipid composition of the fat pad conditioned medium was characterized
using both thin layer and gas liquid chromatography. These lipid analyses indicated that the fat pad pieces released significant
amounts of fatty acids and phospholipids into the medium during the conditioning period. The free fatty acid composition included
both saturated and unsaturated molecules, and about 80% of the total fatty acids consisted of palmitate, stearate, oleate,
and linoleate. These same fatty acids were a structural component of the majority of phospholipid found in the medium. The
addition of palmitate or stearate to defined medium had no effect or was inhibitory for CL-S1 proliferation, depending on
the concentration used. Defined medium supplemented with oleate, arachidonate, or linoleate induced CL-S1 proliferation, and
the inhibitory effects of palmitate and stearate were overcome by addition of oleate and linoleate. These data indicate that
both unsaturated and saturated fatty acids are released from intact adipose cells of the mouse mammary fat pad and that fatty
acids can influence the growth of prenoplastic mouse mammary epithelium. Thus, unsaturated fatty acids, perhaps in conjunction
with other substances released simultaneously, are candidate molecules for the substances that mediate the effect of adipose
tissue on growth of epithelium.
This work was supported in part by a grant from the American Institute for Cancer Research; grant CA 46885 from the National
Institutes of Health, Bethesda, MD; and by State of Washington initiative 171. 相似文献
108.
The influence of kairomones on the numerical response of the parasitoidTrioxys indicus against its hostAphis craccivora at its varying density was studied. The kairomones (applied as aqueous extract of the host) significantly enhanced the rate
of parasitisation and multiplication and the area of discovery of the parasitoid and also the K-values of mortality of the
host at all parasitoid densities introduced (1, 2, 4, 8, 12 and 16 parasitoids) into troughs having about 200 hosts. The sex-ratio
of F1 offspring decreased at lower parasitoid densities and remained more or less unchanged at higher parasitoid densities after
the application of kairomones. The present findings indicate that if kairomones are applied properly, the number of hosts
destroyed by a stimulated parasitoid will be about 200, twice the number reported earlier, thus fewer parasitoids will be
needed to regulate an estimated population of the hosts.
相似文献
109.
Colony-stimulating factor 1 (CSF-1) selectively supports the survival, proliferation, and maturation of hemopoietic cells of the monocyte/macrophage lineage. Although the cellular receptor for CSF-1, (the c-fms protein) is a protein-tyrosine kinase activated by the binding of CFS-1, the role of phosphorylation of cellular proteins in CSF-1 signal transduction is poorly understood. Therefore, we examined the CSF-1-stimulated phosphorylation of cellular proteins in human BeWo choriocarcinoma cell line (known to express the c-fms protein). BeWo cells were metabolically labeled with 32Pi, stimulated with recombinant human CSF-1, and extracted with detergent. Phosphotyrosyl proteins were isolated from detergent extracts by affinity chromatography on a highly specific antibody to phosphotyrosine. Rapid phosphorylation of 170-kd protein, followed closely by the phosphorylation of a 56-kd protein, was observed in response to CSF-1. The 170-kd phosphotyrosyl protein bound to wheat germ agglutinin and was secondarily immunoprecipitated with a specific anti-fms serum, consistent with its identity as the CSF-1 receptor. Although purified human macrophages that proliferate in culture in response to CSF-1 are not generally accessible, CSF-1 did stimulate the phosphorylation of a 56-kd protein in intact mononuclear leukocytes from human peripheral blood. Thus, the BeWo cell line may represent a good model for the study of CSF-1-stimulated cellular protein phosphorylation. 相似文献
110.
Olwin BB 《Cytotechnology》1989,2(4):351-365
Heparin-binding growth factors modulate diverse biological activities including cellular proliferation, cellular differentiation, morphogenesis, and angiogenesis. Biochemical characterization for two members of the heparin-binding growth factor family, acidic and basic fibroblast growth factors, is extensive, while characterization of the remaining five members is forthcoming. Cell surface receptors have been identified for acidic and basic fibroblast growth factors, but little is known concerning their sites of action in vivo or the mechanisms involved in transducing the energy of growth factor binding to a biological response. An understanding of the biological basis for the diversity of the heparin binding growth factor family and the in vivo actions of these factors will prove a major challenge to future research efforts. 相似文献