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211.
212.
Spyridon Vamvakas Wolfgang Dekant Dietmar Schiffmann Dietrich Henschler 《Cell biology and toxicology》1988,4(4):393-403
S-(chloroethyl)-cysteine (CEC) and S-(1,2-dichlorovinyl)cysteine (DCVO) have been proposed as intermediates in the metabolic transformation of the carcinogens 1,2-dichloroethane and 1,1,2-trichloroethylene. We have tested the ability of CEC and DCVC to induce DNA repair and genotoxic effects at the chromosomal level by comparative assessment of unscheduled DNA synthesis induction and micronucleus formation in Syrian hamster embryo fibroblasts. CEC induced a potent and dose-dependent response in both assays, whereas DCVC treatment resulted in a comparatively weak induction of DNA repair and failed to raise micronucleus formation above control rates. Inhibition of cysteine conjugate \gB-lyase diminished the effect of DCVC, but had no influence on the genotoxicity of CEC either in the unscheduled DNA synthesis or micronucleus assay.Abbreviations AOAA
aminooxyacetic acid
- CEC
S-(chloroethyl)-cysteine; \gB-lyase, cysteine conjugate -lyase
- DCE
1,2-dichloroethane
- DCVC
S(1,2-dichlorovinyl)-cysteine
- GSH
glutathione
- HU
hydroxyurea
- IBR
IBR-modified Dulbecco's Eagle's reinforced medium
- MN2
micronuclei/2,000 cells
- 4-NQO
4-nitroquinoline-1-oxide
- SHE
Syrian hamster embryo fibroblasts; 3H-Thd, 3H-thymidine
- TCE
1,1,2-trichloroethylene
- UDS
unscheduled DNA synthesis 相似文献
213.
214.
流行性出血热病毒R22株cDNA克隆及其特异性鉴定 总被引:3,自引:0,他引:3
用家鼠型流行性出血热病毒R22株RNA,经polyA接尾,以Oligo-dT做引物,合成cDNA。用pUC18为载体转染E.coli Mc1061,建立cDNA克隆。再经菌落杂交,选择病毒特异性的5个阳性克隆制成缺口翻译探针,与病毒RNA3个片段进行反杂交,确定RNA片段的特异性。结果表明,3个克隆为中(M)片段的cDNA,另两个分别为大(L)和小(S)片段cDNA。核苷酸序列分析证明,克隆的DNA中含病毒特异的核苷酸序列。 相似文献
215.
测定了3T3细胞、人和大鼠一些组织中DNA拓扑异构酶Ⅰ的活性;估计了核酸内切酶对拓扑酶Ⅰ松弛活性测定的干扰程度;发现增殖组织全细胞抽提液中酶比活高于正常分化组织,而且在异常增殖组织中酶比活的增高更为显著。 相似文献
216.
Development and characterization of continuous avian cell lines depleted of mitochondrial DNA 总被引:6,自引:0,他引:6
Réjean Morais Paul Desjardins Chanta Turmel Karen Zinkewich-Péotti 《In vitro cellular & developmental biology. Plant》1988,24(7):649-658
Summary Populations of quail and chicken cells were treated with ethidium bromide, an inhibitor of mitochondrial DNA replication.
After long-term exposure to the drug, the cell populations were transferred to ethidium bromide (EtdBr)-free medium, and cloned.
Clones HCF7 (quail) and DUS-3 (chicken) were propagated for more than a year, and then characterized. Analysis of total cellular
DNA extracted from these cells revealed no characteristic mitochondrial DNA molecule by Southern blot hybridization of HindIII-
or AvaI-digested total cellular DNA probed with cloned mitochondrial DNA fragments. Reconstruction experiments, where a small
number of parental cells was mixed with HCF7 cells and DUS-3 cells before extraction of total cellular DNA, further strengthen
the notion that the drug-treated cells are devoid of mitochondrial DNA molecules. The cell populations were found to proliferate
at a moderately reduced growth rate as compared to their respective parents, to be auxotrophic for uridine, and to be stably
resistant to the growth inhibitory effect of EtdBr and chloramphenicol. At the ultrastructural level, mitochondria were considerably
enlarged and there was a severe reduction in the number of cristae within the organelles and loss of cristae orientation.
Morphometric analysis revealed a fourfold increase of the mitochondrial profile area along with a twofold decrease of the
numerical mitochondrial profiles. Analysis of biochemical parameters indicated that the cells grew with mitochondria devoid
of a functional respiratory chain. The activity of the mitochondrial enzyme dihydroorotate dehydrogenase was decreased by
95% and presumably accounted for uridine auxotrophy.
This work was supported by a grant from the Medical Research Council of Canada. 相似文献
217.
A rapid fluorometric DNA assay for the measurement of cell density and proliferation in vitro 总被引:4,自引:0,他引:4
Timothy A. McCaffrey Lily A. Agarwal Babette B. Weksler 《In vitro cellular & developmental biology. Plant》1988,24(3):247-252
Summary Many research efforts require the accurate determination of cell density in vitro. However, physical cell counting is inaccurate,
time-intensive and requires removal of the cells from their growth environment, thereby introducing a host of potential artifacts.
The current studies document a very simple method of determining cell density in microtiter wells via DNA-enhanced fluorescence.
Fixed cells are stained with the A-T intercalating DNA stains DAPI or Hoechst 33342 and then fluorescence is quantified in
a plate fluorometer. Fluorescence is shown to be linearly related to cell density as determined by two physical counting methods.
The validity of the method is established in determining serum-stimulated growth of smooth muscle cells and in mitogen-induced
growth of endothelial cells. The fixed cells can be stored for prolonged periods, thus allowing time-course proliferation
assays without interassay variations. The fixed cells are also suitable for determinations of antigens of interest by ELISA.
This method is potentially valuable in many in vitro systems where the quantification of cell density and proliferation is
necessary.
This work supported in part by NIH Cardiovascular Training Grant HL07423 and a grant from the American Federation for Aging
Research to T. M. and HL35724 to B. W.
EDITOR’S STATEMENT The technique described in this paper represents an approach to quantifying cell density in adherent monolayers
of cultured cells in microtiter wells that is rapid and simple and does not require radioisotopes or removal of cells. 相似文献
218.
Paul H. Gumerlock Benjamin F. Edwards Arline D. Deitch Frederick J. Meyers 《In vitro cellular & developmental biology. Plant》1988,24(5):429-434
Summary A human cell line has been established from a renal adenocarcinoma rib metastasis of a 58-y-old male. This cell line has been
maintained in continuous culture for 20 mo. through more than 50 passages. It displays simulataneous expression of the intermediate
filaments cytokeratin and vimentin. Flow cytometric analysis of DNA content reveals a major hyperdiploid population.
This work was supported in part by a grant from Triton Biosciences, Inc. 相似文献
219.
β-lactam antibiotics in the presence of certain metal ions damage deoxyribose and DNA with the release of thiobarbituric acid-reactive material. This damage can be substantially prevented by catalase, metal chelators and some scavengers of the hydroxyl radical. Ferric salts in the presence of certain β-lactam antibiotics were effective in degrading deoxyribose but they did not appear to damage DNA. In contrast copper salts and p-lactam antibiotics were extremely effective in damaging both DNA and deoxyribose. 相似文献
220.
猴脑线粒体DNA提取及限制性内切酶分析 总被引:2,自引:0,他引:2
本文用冷碱法从2只恒河猴和1只食蟹猴的脑组织中提取mtDNA,最后的得率大约为0.7μgmtDNA/g脑组织,是肝脏组织得率的1/3左右。与肝脏组织相比较,从脑组织中提取mtDNA有以下优点:1.匀浆方便。2.样品中蛋白杂质少,容易彻底抽提去除蛋白质。3.大分子RNA杂质极少,不经Sepharose-4B柱或RNawe处理,就可得到较纯的样品。加之哺乳动物脑的体积较大,哺乳动物脑组织不失为提取mtDNA的一个有用的组织来源。经16种限制性内切酶分析,并与来自同一个体肝脏组织的mtDNA比较,结果进一步证实,mtDNA无组织特异性。对12岁以上老年猴脑mtDNA的分析表明,在衰老中,动物mtDNA的序列可能没有变化,甲基化程度也无显著增高。 相似文献