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991.
992.
Daniel E. Gomez Jacqueline L. Hartzler Robert H. Corbitt Alexander M. Nason Unnur P. Thorgeirsson 《In vitro cellular & developmental biology. Animal》1993,29(6):451-455
Summary We describe a fast and reproducible method that can be used as a final step in obtaining pure populations of liver endothelial
cells. This method employs endothelial cell specific lectin covalently bound to magnetic polystyrene beads (Dynabeads). Evonymus
europaeus agglutinin (EEA)-coated Dynabeads were used to purify monkey liver endothelium from Percoll gradient separated nonparenchymal
cells. EEA-coated beads were also successfully used to purify monkey aortic endothelial cells. The endothelial cells grew
to confluence as a cobblestonelike monolayer, expressed Factor VIII related antigen, and incorporated acetylated-low density
lipoprotein. The magnetic beads seemed not to modify the normal properties of the isolated endothelium, thus facilitating
their use in experimental studies. This immunomagnetic separation technique may be applicable for purification of endothelial
cells from a wide variety of tissue sources. 相似文献
993.
Johanna Plendl Laura Hartwell Robert Auerbach 《In vitro cellular & developmental biology. Animal》1993,29(1):25-31
Summary Endothelial cells of the NMRI mouse strain express a cell surface glycoprotein recognized by the lectinDolichos biflorus agglutinin (DBA). This study documents a marked organ-specific increase in DBA-specific lectin binding of myocardium-derived
endothelial cells (MEC) of the NMRI/GSF mouse during in vitro cultivation. An up to 20-fold increase in DBA binding sites
is observed in long-term culture, an increase not found in other NMRI-derived endothelial cell lines (e.g., brain, aorta).
The increase appears restricted to DBA in that binding with other lectins (PNA, WGA) was unaltered. NMRI MEC cultures maintain
typical endothelial cell attributes such as cobblestone morphology on confluence, expression of endothelial cell-specific
surface markers, and production of angiotensin-converting enzyme. Cultures routinely become aneuploid within 4 passages, several
passages before upregulation of the DBA binding site(s). Myocardial endothelial cells sorted to obtain DBAhi and DBAlo cell populations generally maintained their sorted phenotype for 3 to 4 passages. Limiting dilution cloning resulted in clones
varying in DBA expression. Clones for DBAhi expression maintained their DBA affinity for at least 10 passages (>30 doublings), whereas DBAlo clones gave rise to varying numbers of DBAhi cells within 2 to 4 passages. We hypothesize that the change in DBA affinity accompanies in vitro aging, that the change
is independent of alterations in karyotype, and that the increase in DBA affinity may reflect a change in one or more other
endothelial cell properties. Additional studies will be necessary to determine whether the in vitro changes are correlated
with specific functional alterations and whether they accurately reflect progressive changes of MEC in vivo. 相似文献
994.
Alan M. Goldberg John M. Frazier David Brusick Michael S. Dickens Oliver Flint Stephen D. Gettings Richard N. Hill Robert L. Lipnick Kevin J. Renskers June A. Bradlaw Robert A. Scala Bellina Veronesi Sidney Green Neil L. Wilcox Rodger D. Curren 《In vitro cellular & developmental biology. Animal》1993,29(9):688-692
Summary The development and application of in vitro alternatives designed to reduce or replace the use of animals, or to lessen the
distress and discomfort of laboratory animals, is a rapidly developing trend in toxicology. However, at present there is no
formal administrative process to organize, coordinate, or evaluate validation activities. A framework capable of fostering
the validation of new methods is essential for the effective transfer of new technologic developments from the research laboratory
into practical use. This committee has identified four essential validation resources: chemical bank(s), cell and tissue banks,
a data bank, and reference laboratories. The creation of a Scientific Advisory Board composed of experts in the various aspects
and endpoints of toxicity testing, and representing the academic, industrial, and regulatory communities, is recommended.
Test validation acceptance is contingent on broad buy-in by disparate groups in the scientific community—academics, industry,
and government. This is best achieved by early and frequent communication among parties and agreement on common goals. It
is hoped that the creation of a validation infrastructure composed of the elements described in this report will facilitate
scientific acceptance and utilization of alternative methodologies and speed implementation of replacement, reduction, and
refinement alternatives in toxicity testing. 相似文献
995.
996.
Maria Ermakova Russell Woodford Zachary Taylor Robert T. Furbank Srinivas Belide Susanne von Caemmerer 《Plant biotechnology journal》2023,21(6):1206-1216
Sorghum is one of the most important crops providing food and feed in many of the world's harsher environments. Sorghum utilizes the C4 pathway of photosynthesis in which a biochemical carbon-concentrating mechanism results in high CO2 assimilation rates. Overexpressing the Rieske FeS subunit of the Cytochrome b6f complex was previously shown to increase the rate of photosynthetic electron transport and stimulate CO2 assimilation in the model C4 plant Setaria viridis. To test whether productivity of C4 crops could be improved by Rieske overexpression, we created transgenic Sorghum bicolor Tx430 plants with increased Rieske content. The transgenic plants showed no marked changes in abundances of other photosynthetic proteins or chlorophyll content. The steady-state rates of electron transport and CO2 assimilation did not differ between the plants with increased Rieske abundance and control plants, suggesting that Cytochrome b6f is not the only factor limiting electron transport in sorghum at high light and high CO2. However, faster responses of non-photochemical quenching as well as an elevated quantum yield of Photosystem II and an increased CO2 assimilation rate were observed from the plants overexpressing Rieske during the photosynthetic induction, a process of activation of photosynthesis upon the dark–light transition. As a consequence, sorghum with increased Rieske content produced more biomass and grain when grown in glasshouse conditions. Our results indicate that increasing Rieske content has potential to boost productivity of sorghum and other C4 crops by improving the efficiency of light utilization and conversion to biomass through the faster induction of photosynthesis. 相似文献
997.
998.
Robert M. Mader Blanka Rizovski Günther G. Steger Hugo Rainer Reinelde Proprentner Rainer Kotz 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》1993,613(2)
An high-performance liquid chromatographic method with column switching for the detection of less than 4 ng of methotrexate in the urine of oncologic nurses is described. Urine samples were purified by solid-phase extraction on silica-bonded phenyl columns, eluting impurities with ethyl acetate. After elution from the column, the analyte was concentrated ten-fold, evaporating the solvent. On a strong anion-exchange column (Nucleosil 100 SB), methotrexate was separated from the remaining interfering substances, was then switched to a reversed-phase column (LiChrospher 100 RP-18e), and finally eluted by a linear gradient in a solvent system consisting of ammonium formate buffer (pH 2.7) and acetonitrile. Absorbance was monitored at 310 nm. This method has proved to be suitable for detecting traces of methotrexate in urine in order to individualize risks and to reduce further the occupational safety hazard for hospital personnel. 相似文献
999.
1000.