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31.
Identification of a Thymidylate Synthase Gene within the Genome of Chilo Iridescent Virus 总被引:1,自引:0,他引:1
The thymidylate synthase (TS, EC 2.1.1.45) is essential for the de novo synthesis of dTMP in pro- and eucaryotic organisms. Consequently it plays a major role in the replication of the DNA genome of a cell or a DNA virus. The gene encoding the TS of Chilo iridescent virus (CIV) was identified by nucleotide sequence analysis of the viral genome and was mapped within the EcoRI CIV DNA fragments G and R. Computer assisted analysis of the DNA nucleotide sequence between the genome coordinates 0.482 and 0.489 revealed an open reading frame (ORF) of 885 nucleotides. This ORF was found to encode a polypeptide of 295 amino acid residues (33.9 kDa) that showed significant homologies to known TS of different species including mammals, plants, fungi, protozoa, bacteria, and DNA viruses. The highest amino acid homologies were found between the CIV-TS and the TS of herpesvirus ateles (54.0%), Saccharomyces cerevisiae (51.8%), herpesvirus saimiri (51.0%), rhesus monkey rhadinovirus (50.7%), mouse (50.5%), rat (50.2%), varicella-zoster virus (50.2%), equine herpesvirus 2 (50.0%), and the human TS (48.4%). The CIV-TS contains six amino acid domains that are highly conserved in the TS of other species. Within these domains the major amino acid residues are present for which a functional role has been reported. The CIV-TS was found to be more closely related to the TS of eucaryotes than to the TS of procaryotes indicating the phylogenetic origin of the CIV-TS gene. The identification of a TS gene in the genome of CIV is the first report of a viral TS that is not encoded by a herpesvirus or a bacteriophage. 相似文献
32.
Stessman J Maaravi Y Hammerman-Rozenberg R Cohen A Nemanov L Gritsenko I Gruberman N Ebstein RP 《Mechanisms of ageing and development》2005,126(2):333-339
In an exploratory study, 11 common polymorphisms were examined for contributing to longevity including: apolipoprotein E (apoE), methylenetetrahydrofolate reductase (MTHFR), cathepsin D (CAD), superoxide dismutase 2 (SOD2), angiotensinogen (AGT) and insulin-like growth factor 2 (IGF2), Leiden factor 7, p53 oncogene, dopamine D4 receptor (DRD4) and the serotonin transporter (SERT). Genotype and allele frequencies of these genes were compared in 224 older (75 years) Jewish Jerusalem residents of Ashkenazi ethnicity to a group of 441 younger subjects (22 years). Nominally significant results provide suggestive evidence in the Ashkenazi group that apoE, MHTFR, SOD2, IGF2 ApaI, and factor VII are risk factors for a single outcome, survival to 75. Overall, the more genetically homogenous Ashkenazi ethnic group showed evidence for association in five genes examined suggesting that future studies in this population would gainfully focus on this ethnic group. 相似文献
33.
We describe a protocol for the synchronisation of normal and tumour cells grown in suspension cultures using 3-hydroxypyridin-4-one iron chelators. These compounds inhibit ribonucleotide reductase, one of the rate limiting enzymes in DNA synthesis, and so block the cell cycle in late G1 phase. After removal of the chelator or repletion of cellular iron, cells progress through the cycle and remain synchronised for at least one full cell cycle. Cell viability is unaffected for at least 72 hours post-incubation and chelator treatment has no effect on RNA and protein synthesis. This method of synchronisation has been successful with all cell lines tested including normal and leukaemic human cell lines. 相似文献
34.
Summary The phenotypically wild strain I3 of Chlamydomonas reinhardtii, carrying a cryptic mutation at the nit-6 locus, was distinguished from strains 21gr (cryptic mutant at nit-5) and 6145c (wild type) because of the ability of I3 to grow on nitrate media containing 2mM tungstate.Molybdopterin-cofactor (MoCo) mutants 102 (double mutant at nit-5 and nit-6) and 104 (mutant at nit-4) grew on nitrate media supplemented with high concentrations of molybdate, although final cell densities were 40–60% lower and generation times 3.5 to six fold longer than for wild type. Under these conditions, nitrate reductase (NR) activity of the mutants, when measured either in situ or in vitro, was practically undetectable. The MoCo defective mutant 307 (nit-3) was not molybdate repairable. Although NR activity was not restored in vitro by molybdate in any of the MoCo– mutant strains, their extracts had free NR-diaphorase subunits together with NR-subunits assembled into high molecular weight species.Our results indicate that: a) nit-4, nit-5 and nit-6 loci are responsible for molybdate processing in the cell; b) nit-3 may encode a component of the pterin moiety biosynthetic route; c) NR subunits can assemble in the presence of an inactive MoCo; d) high concentrations of molybdate can replace partially in vivo but not in vitro the function of nit-4 and the combined function(s) of the nit-5 and nit-6 gene products. 相似文献
35.
36.
Cloning of the nitrate reductase gene of Stagonospora (Septoria) nodorum and its use as a selectable marker for targeted transformation 总被引:1,自引:0,他引:1
The nitrate reductase gene (NIA1) of the phytopathogenic fungus Stagonospora (Septoria) nodorum has been cloned from a cosmid library by homologous hybridisation with a PCR-generated probe. A 6.7-kb fragment carrying
the NIA1 gene was subcloned and partially characterised by restriction mapping. Sequencing of the gene indicated a high degree of
homology, both at the nucleotide and amino-acid levels, with nitrate reductase genes of other filamentous fungi. Furthermore,
consensus regulatory signals thought to be involved in the control of nitrogen metabolism are present in the 5′ flanking region.
The cloned NIA1 gene has been used to develop a gene-transfer system based on nitrate assimilation. Stable nia1 mutants of S. nodorum defective in nitrate reductase were isolated by virtue of their resistance to chlorate. These were transformed back to nitrate
utilisation with the wild-type S. nodorum NIA1 gene. Southern analyses revealed that transformation occurred as a result of the integration of transforming DNA into the
fungal genome; in all cases examined, integration was targeted to the homologous sequence.
Received: 30 March / 9 June 1998 相似文献
37.
Liu CS Chen HW Lii CK Tsai CS Kuo CL Wei YH 《Environmental and molecular mutagenesis》2002,40(3):168-174
The effects of long-term smoking on mitochondrial DNA (mtDNA) deletions in hair follicles were investigated in subjects with different antioxidant capacity. Twenty-two male smokers with a smoking index of greater than 5 pack-years and without any known systemic diseases were recruited for this study. Forty healthy nonsmoking males were included as controls. We found that the concentrations of ascorbate and alpha-tocopherol and the activities of glutathione S-transferase (GST) and glutathione peroxidase in blood plasma were significantly decreased in smokers. The levels of glutathione and protein thiols in whole blood and the incidence of a 4,977 bp deletion of mtDNA (dmtDNA) in hair follicles were significantly increased in smokers. A significantly higher incidence of the 4,977 bp dmtDNA was found in smokers with plasma GST activity less than 5.66 U/l (OR = 7.2, P = 0.020). Using multiple covariate ANOVA and logistic regression, we found that age and low plasma GST activity were the only two risk factors for the 4,977 bp dmtDNA. These results suggest that smoking depletes antioxidants and causes mtDNA deletions and that plasma GST may play an important role in the preservation of the mitochondrial genome in tissue cells of smokers. 相似文献
38.
39.
F. V. Donenko S. M. Sitdikova A. O. Kabieva B. E. Polotskii Z. O. Machaladze M. I. Davydov L. V. Moroz 《Bulletin of experimental biology and medicine》1995,119(2):206-208
Preincubation of cells of BDF1 hybrid mice with P388 leukemia with doxorubicin and buthionine sulfoximine leads to the manifestation
of a therapeutic effect of the antibiotic. Injection of buthionine sulfoximine and ethacrinic acid to mice with leukemia does
not alter the therapeutic effect of the antibiotic.
Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 119, N
o
2, pp. 212–214, February, 1995
Presented by N. N. Trapeznikov, Member of the Russian Academy of Medical Sciences 相似文献
40.
P糖蛋白和谷胱甘肽—S—转移酶在肾细胞癌中的表达 总被引:2,自引:1,他引:2
探讨肾细胞癌与多药耐药的关系,方法:应用免疫组化方法,检测4例术前未进行化疗的肾细胞癌和16例正常肾组织中的P糖蛋白和谷胱甘肽-2转移-π表达。结果:P-ag和GST-π外地16例正常肾组织中的表达率均匀100%,在44例肾细胞癌组织中分别为63.65和54.5%Pg,P-gp和/或GST-π阳性表达率为79.5%。 相似文献