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101.
Socrates J. Tzartos Heleni V. Loutrari Fen Tang Anna Kokla Susan L. Walgrave Robert P. Milius† Bianca M. Conti-Tronconi 《Journal of neurochemistry》1990,54(1):51-61
Most anti-nicotinic acetylcholine receptor (AChR) antibodies in myasthenia gravis are directed against an immunodominant epitope or epitopes [main immunogenic region (MIR)] on the AChR alpha-subunit. Thirty-two synthetic peptides, corresponding to the complete Torpedo alpha-subunit sequence and to a segment of human muscle alpha-subunit, were used to map the epitopes for 11 monoclonal antibodies (mAbs) directed against the Torpedo and/or the human MIR and for a panel of anti-AChR mAbs directed against epitopes on the alpha-subunit other than the MIR. A main constituent loop of the MIR was localized within residues alpha 67-76. Residues 70 and 75, which are different in the Torpedo and human alpha-subunits, seem to be crucial in determining the binding profile for several mAbs whose binding to the peptides correlated very well with their binding pattern to native Torpedo and human AChRs. This strongly supports the identification of the peptide loop alpha 67-76 as the actual location of the MIR on the intact AChR molecule. Residues 75 and 76 were necessary for binding of some mAbs and irrelevant for others, in agreement with earlier suggestions that the MIR comprises overlapping epitopes. Structural predictions for the sequence segment alpha 67-76 indicate that this segment has a relatively high segmental mobility and a very strong turning potential centered around residues 68-71. The most stable structure predicted for this segment, in both the Torpedo and human alpha-subunits, is a hairpin loop, whose apex is a type I beta-turn and whose arms are beta-strands. This loop is highly hydrophilic, and its apex is negatively charged. All these structural properties have been proposed as characteristic of antibody binding sites. We also localized the epitopes for mAbs against non-MIR regions. Among these, the epitope for a monoclonal antibody (mAb 13) that noncompetitively inhibits channel function was localized within residues alpha 331-351. 相似文献
102.
Daniel K. Yee Anna Pastuszko David Nelson David F. Wilson 《Journal of neurochemistry》1989,52(1):54-60
Incubation of synaptosomes from rat brain with DL-2-amino-5-phosphonovalerate (APV) stimulated an increased release of dopamine, and this effect was strictly dependent on the extrasynaptosomal calcium level. APV increased biosynthesis of dopamine from tyrosine by 30%, whereas monoamine oxidase activity was inhibited by 30%. When synaptosomes were incubated with radioactive dopamine, APV caused a large decrease in incorporation of label into 3,4-dihydroxyphenylacetic acid but greatly increased incorporation into norepinephrine and its N-methyl derivatives. Quantification of dopamine and its metabolites in synaptosomes, using electrochemical detection, indicated that the presence of APV resulted in changes in the absolute levels of the aforementioned dopamine metabolites similar to the changes in radiolabel incorporation. Omission of Ca2+ from the extrasynaptosomal medium greatly diminished the APV-induced changes in catecholamine metabolism. The metabolic changes appear to largely result from an increased intrasynaptosomal Ca2+ level due to the APV-induced increase in calcium permeability of the plasma membrane. 相似文献
103.
Anna L. Trifillis Myong Won Kahng 《In vitro cellular & developmental biology. Plant》1990,26(5):441-446
Summary We have developed an in vitro model of human papillary collecting duct cells isolated from cadaver kidneys using methods similar
to those we previously reported for the isolation of human proximal tubule cells. To date we have isolated papillary collecting
duct cells from 100 normal human kidneys. Papillae were dissected and digested in Cellgro containing 400 U/ml collagenase.
Cells were plated on fibronectin-coated culture flasks at a density of 104 live cells/ml in Cellgro supplemented with insulin and 10% fetal bovine serum. Confluent monolayers, which were able to withstand
600 mOsm for 8 h, were obtained within 10 to 15 d. Cells of primary isolates and first passages exhibited epithelial cell ultrastructure
including cell junctions, microvilli, and cilia. A dark-brown reaction product was observed in these cells when stained by
the immunoperoxidase method with peroxidase-labeled peanut lectin (Arachis hypogaea), which binds specifically to human distal tubule and collecting duct cells. These cells were negative for Factor-VIII (a
marker for endothelial cells) and γ-glutamyltransferase (a marker for proximal tubule cells). High activities of the glycolytic
enzyme pyruvate kinase and arginine vasopressin-stimulated cAMP production in these cells are consistent with a distal nephron
origin. The results indicate that human collecting duct cells can be isolated and cultured to provide an in vitro system to
probe pathogenetic mechanisms of potential nephrotoxins.
Part of this work was presented at a Symposium of the Center for Alternatives to Animal Testing, April 4–5, 1989, Johns Hopkins
Medical Institutions, Baltimore, MD 21205.
This work was supported in part by grants R01-AI24179, PO1-A804393 for the Public Health Service, U.S. Department of Health
and Human Services, and by a grant from the National Kidney Foundation, Baltimore, MD affiliate. 相似文献
104.
105.
U Danielsson 《Journal of applied physiology》1990,68(3):1275-1281
A heat flux disk has been developed that directly measures the convective heat transfer in W/m2. When the sensor is calibrated on an aluminum cylinder, the calibration constant obtained is greatest in still air. As air movement increases, the calibration constant is reduced with increasing convective heat transfer coefficient, 0.5%.W-1.m2.K. The influence of wind on the calibration value is greatly reduced when the sensor is attached to a surface with lower thermal conductivity. The local convective heat transfer coefficient (hc) of the human body was measured. The leg acts in a manner similar to that of a cylinder, with the highest hc value at the front facing the wind and the lowest approximately 90 degrees from the wind, and in the wake a value is obtained that is close to the average hc value of the leg. When hc is measured at several angles and positions all over the body, the results indicate that the body acts approximately as a cylinder with a hc value related to the wind speed as hc = 8.6.v0.6 W.m-2.K-1, where v is velocity. 相似文献
106.
Anna Jachner 《Hydrobiologia》1996,325(2):151-155
The response of bleak (Alburnus alburnus (L.)) to alarm substance (from skin extract of conspecifics) was quantified, using a video camera, in laboratory experiments across two light and one dark periods (18 hrs). In the light alarm substance induced hiding in refuges (vegetation) and formation of aggregations. There was no such reaction during darkness or during a second light period. 相似文献
107.
Butomus umbellatus L. is a plant species typical of littoral communities of river and stream shores. It can form continuous stands in shallow reservoirs with fluctuating water level. Their expansion is promoted by: (a) intensive vegetative reproduction of plants, (b) crowded sprouting from rhizome fragments on emerged pond bottom, (c) shallow water layer in the year following summer drainage. Expansion of B. umbellatus depends on ploidy level: two cytotypes were found in the Czech and Slovak Republics, differing in their reproductive ability. Seed production of triploids is strongly limited (they are self-incompatible within clones), while diploids can be fully fertile. Nevertheless, even in diploids, the efficiency of seed reproduction under natural conditions is low. Triploids spread by intensive vegetative reproduction, which is decisive for clonal growth of populations and their regeneration after scraping of bottom surface. During seasonal development, maximum of aboveground biomass is produced in early summer, while underground biomass increases till autumn. Growth of the plants is limited by cutting before maximum underground biomass is attained, or by duck grazing. 相似文献
108.
Further studies on theisfA mutation responsible for anti-SOS and antimutagenic activities inEscherichia coli are described. We have previously shown that theisfA mutation inhibits mutagenesis and other SOS-dependent phenomena, possibly by interfering with RecA coprotease activity. TheisfA mutation has now been demonstrated also to suppress mutator activity inE. coli recA730 andrecA730 lexA51(Def) strains that constitutively express RecA coprotease activity. We further show that the antimutator activity of theisfA mutation is related to inhibition of RecA coprotease-dependent processing of UmuD. Expression of UmuD' from plasmid pGW2122 efficiently restores UV-induced mutagenesis in therecA730 isfA strain and partially restores its mutator activity. On the other hand, overproduction of UmuD'C proteins from pGW2123 plasmid markedly enhances UV sensitivity with no restoration of mutability. 相似文献
109.
Marcella Carcupino Anna Maria Fausto Maria Luisa Bernardino Ortega Marzio Zapparoli Massimo Mazzini 《Zoomorphology》1996,116(3):103-110
Spermatophore development and ultrastructure of the mature sperm of Craterostigmus tasmanianus were studied using light and electron microscopy. In C. tasmanianus, as in the Scolopendromorpha, the spermatophore develops within the vas deferens. The latter consists of three parts, each
with a different morphology. The first may be involved in guiding the sperm to roll up into typical ring-like structures,
while the other two, which show an evident secretory activity, secrete the acellular wall of the spermatophores. The ultrastructure
of mature spermatozoa showed that a very close similarity exists between Craterostigmomorpha and Lithobiomorpha, especially
regarding the organization of the connecting piece. Based on this similarity, we consider the Craterostigmomorpha together
with the Scolopendromorpha, Geophilomorpha and Lithobiomorpha (=Pleurostigmophora) to be the sister group of the Scutigeromorpha.
Accepted: 2 June 1996 相似文献
110.
In this report we investigate the inhibition of membrane conductivity, due to the murine polyomavirus infection in permissive cells in culture. We define experimental conditions to have reproducible results and demonstrate that the intensity of the effects on the cell membrane, depends upon the virus titer used in the infection. Finally, the virus dependent effects disappear if the infection is performed in the presence of a drug that inhibits polymavirus DNA replication. 相似文献