全文获取类型
收费全文 | 6243篇 |
免费 | 577篇 |
国内免费 | 3篇 |
专业分类
6823篇 |
出版年
2023年 | 28篇 |
2022年 | 62篇 |
2021年 | 122篇 |
2020年 | 65篇 |
2019年 | 85篇 |
2018年 | 109篇 |
2017年 | 98篇 |
2016年 | 162篇 |
2015年 | 277篇 |
2014年 | 317篇 |
2013年 | 402篇 |
2012年 | 429篇 |
2011年 | 471篇 |
2010年 | 281篇 |
2009年 | 265篇 |
2008年 | 386篇 |
2007年 | 377篇 |
2006年 | 312篇 |
2005年 | 333篇 |
2004年 | 311篇 |
2003年 | 315篇 |
2002年 | 309篇 |
2001年 | 46篇 |
2000年 | 30篇 |
1999年 | 50篇 |
1998年 | 76篇 |
1997年 | 49篇 |
1996年 | 53篇 |
1995年 | 39篇 |
1994年 | 44篇 |
1993年 | 47篇 |
1992年 | 37篇 |
1991年 | 34篇 |
1990年 | 27篇 |
1989年 | 29篇 |
1988年 | 22篇 |
1987年 | 29篇 |
1983年 | 27篇 |
1982年 | 35篇 |
1981年 | 25篇 |
1980年 | 24篇 |
1979年 | 26篇 |
1978年 | 30篇 |
1976年 | 20篇 |
1974年 | 23篇 |
1973年 | 21篇 |
1972年 | 26篇 |
1970年 | 20篇 |
1967年 | 28篇 |
1966年 | 20篇 |
排序方式: 共有6823条查询结果,搜索用时 31 毫秒
21.
Improved media for normal human muscle satellite cells: Serum-free clonal growth and enhanced growth with low serum 总被引:5,自引:0,他引:5
Richard G. Ham Judy A. St. Clair Cecelia Webster Helen M. Blau 《In vitro cellular & developmental biology. Plant》1988,24(8):833-844
Summary We have developed a serum-free medium for clonal growth of normal human muscle satellite cells (HMSC). It consists of an optimized
nutrient medium MCDB 120, plus a serum-free supplement, designated SF, that contains epidermal growth factor (EGF), insulin,
dexamethasone, bovine serum albumin, and fetuin. Fibroblast growth factor was needed with dialyzed fetal bovine serum (dFBS)
as the only other supplement, but in media containing SF, it was only slightly beneficial, and was omitted from the final
medium without significant loss. Clonal growth of HMSC in MCDB 120 plus SF is as good as with 15% serum and 0.5% chicken embryo
or bovine pituitary extract. However, growth is further improved by use of a doubly-supplemented (DS) medium containing both
SF and 5% dFBS. Clonal growth of HMSC in the DS medium far exceeds that in previous media with any amount of serum, and monolayer
growth is at least equal to that in conventional media with higher levels of serum. Cells grown in these media exhibit little
differentiation, even when grown to high densities. However, they retain the capacity for extensive fusion and synthesis of
increased creatine kinase when transferred to a serum-free differentiation-promoting medium, such as Dulbecco's modified Eagle's
medium plus insulin. All experiments were done with clonal cultures of HMSC to insure that observed growth responses were
always those of muscle cells.
This research was supported by a grant from the Muscular, Dystrophy Association.
Editor's statement This article describes the optimization of both the basal nutrient medium and growth factor requirements
for human muscle cells in vitro. This system is critical for studies of normal muscle cell and molecular biology, as well
as for understanding diseases of muscle such as Duchenne, Muscular Dystrophy. 相似文献
22.
Mapping of FMR1, the gene implicated in fragile X-linked mental retardation, on the mouse X chromosome 总被引:2,自引:0,他引:2
A genetic map of the Cf-9 to Dmd region of the mouse X chromosome has been established by typing 100 offspring from a Mus musculus x Mus spretus interspecific backcross for the four loci Cf-9, Cdr, Gabra3, and Dmd. The following order and genetic distances in centimorgans were determined: (Cf-9)-2.4 +/- 1.7-(Cdr)-2.0 +/- 1.4-(Gabra3)-4.1 +/- 2.0-(Dmd). Six backcross offspring carrying X chromosomes with recombination events in the Cdr-Dmd region were identified. These recombination events were used to define the position of Fmr-1, the murine homologue of FMR1, which is the gene implicated in the fragile X syndrome in man, and that of DXS296h, the murine homologue of DXS296. Both Fmr-1 and DXS296h were mapped into the same recombination interval as Gabra3 on the mouse X chromosome. These findings provide strong support for the concept that the order of loci lying in the Cf-9 to Gabra3 segment of the X chromosome is highly conserved between human and mouse. 相似文献
23.
We describe here a method for DNA fingerprinting of human chromosomes by Alu-polymerase chain reaction (PCR) amplification of DNA from monochromosomal hybrids, following digestion with restriction endonucleases. DNA digestion with restriction enzymes prior to PCR amplification reduces the total number of amplified fragments. The number and pattern of bands of PCR products observed in an electrophoretic medium are chromosome specific and provide a "fingerprint signature" for individual human chromosomes. Using this approach, we have produced fingerprints for human chromosomes 2, 5, 7, 9, and 12. The applicability of this approach to chromosome identification was assessed by comparing the fingerprints obtained for two different hybrids containing chromosome 7. DNA fragments specific for the long and the short arms of human chromosome 12 have also been identified. In addition, Alu-PCR-generated DNA fragments, specific for different chromosomes, were used to probe Southern blots of a hybrid cell panel to identify human chromosomes present in hybrid cell lines. The chromosomal specificity of these probes permits the identification of intact as well as rearranged chromosomes composed of segments arising from more than one chromosome. 相似文献
24.
The cytogenetic effects of the antimetabolite, cytosine arabinoside (Ara-C) are evaluated using in vivo and in vitro test systems and applying multiple parameters. The in vivo assay was carried out on 8-10-week-old inbred Swiss albino male mice using bone marrow as the somatic test system and the cells of testis as the meiotic test system. In vitro human leukocyte cultures were also employed. In vivo experimental doses were computed on surface area basis within the therapeutic dose range and injected intraperitoneally and for in vitro they were calculated on blood volume basis. Evaluation of somatic chromosome mutations included conventional screening for chromosome aberrations, variations in mitotic index and sister-chromatid exchanges (SCEs) by in vivo and in vitro methods besides studies on meiotic test systems using conventional screening for chromosome and sperm-head abnormalities. The quantitative data were subjected to statistical analysis by applying appropriate tests to evaluate their significance. The results of in vivo and in vitro experiments reveal the chromosome mutational activity of the compound. This is further supported by data on SCEs from both systems. However, a comparison of both demonstrated a differential mutagenic response of the drug, more in vivo than in vitro. This is also true for SCEs. Even though the mechanisms involved in causing chromosome aberrations and SCEs are different, the data on both corroborate each other on induction of chromosome mutations. 相似文献
25.
Helen Wiseman Michael Cannon Henry R.V. Arnstein David J. Barlow 《生物化学与生物物理学报:疾病的分子基础》1992,1138(3):197-202
The anti-cancer drug tamoxifen is a potent inhibitor of lipid peroxidation induced by Fe(III)-ascorbate in ox-brain phospholipid liposomes. Similar anti-oxidant effects, but with varying potencies, are also shown by 4-hydroxytamoxifen, cholesterol, ergosterol and 17-β-oestradiol. We now describe a computer-graphic fitting technique that demonstrates a structural similarity between the five compounds. In addition, we have quantified the differences (relative to cholesterol) between the anti-oxidant activities of the compounds in terms of a novel expression reffered to here as the cholesterol coefficient (Cc) Finally, we discuss how the inhibitory effect of tamoxifen on lipid peroxidation may result from a membrane stabilization that is associated with a decrease in membrane fluidity. This action may be related to the anti-proliferative effect exerted by tamoxifen on cancer and fungal cells. 相似文献
26.
Summary We have devised techniques to culture whole, dissected embryos of Drosophila melanogaster. We examine multiple aspects of the morphological and physiological development of the epidermis, musculature, nervous system, and internal organs in this cultured preparation, and show that in vitro development closely parallels normal embryogenesis. These techniques permit a wide range of experimental manipulations during embryogenesis and allow us to extend observations through late embryonic stages, after cuticle deposition. Applications of this technique are presented. 相似文献
27.
A homologue of the Escherichia coli DsbA protein involved in disulphide bond formation is required for enterotoxin biogenesis in Vibrio cholerae 总被引:22,自引:0,他引:22
A strain of Vibrio cholerae, which had been engineered to express high levels of the non-toxic B subunit (EtxB) of Escherichia coli heat-labile enterotoxin, was subjected to transposon (TnphoA) mutagenesis. Two chromosomal TnphoA insertion mutations of the strain were isolated that showed a severe defect in the amount of EtxB produced. The loci disrupted by TnphoA in the two mutant derivatives were cloned and sequenced, and this revealed that the transposon had inserted at different sites in the same gene. The open reading frame of the gene predicts a 200-amino-acid exported protein, with a Cys-X-X-Cys motif characteristic of thioredoxin, protein disulphide isomerase, and DsbA (a periplasmic protein required for disulphide bond formation in E. coli). The V. cholerae protein exhibited 40% identity with the DsbA protein of E. coli, including 90% identity in the region of the active-site motif. Introduction of a plasmid encoding E. coli DsbA into the V. cholerae TnphoA derivatives was found to restore enterotoxin formation, whilst expression of Etx or EtxB in a dsbA mutant of E. coli confirmed that DsbA is required for enterotoxin formation in E. coli. These results suggest that, since each EtxB subunit contains a single intramolecular disulphide bond, a transient intermolecular interaction with DsbA occurs during toxin subunit folding which catalyses formation of the disulphide in vivo. 相似文献
28.
Jeong-Yau Ho Rob Weide Helen M. Ma Monique F. van Wordragen Kris N. Lambert Maarten Koornneef Pim Zabel Valerie M. Williamson 《The Plant journal : for cell and molecular biology》1992,2(6):971-982
A dominant allele at the Mi locus on chromosome 6 of tomato (Lycopersicon esculentum Mill) confers resistance to three species of root-knot nematodes (Meloidogyne). The resistance, which is associated with a localized necrotic response, was originally introduced into tomato from the wild species Lycopersicon peruvianum. As a step towards the molecular cloning of Mi, we have identified closely linked DNA markers from both cDNA and genomic DNA libraries as restriction fragment length polymorphisms (RFLPs). DNA from tomato populations segregating for nematode resistance was analyzed to generate a high-resolution genetic map of this region. Additional information on gene order was obtained by comparing the size of the introgressed L. peruvianum chromosomal segment within a collection of nematode-resistant tomato lines. Among the four cDNA markers that are tightly linked to Mi, three are dominant, i.e. L. peruvianum-specific. One cDNA marker corresponds to a gene family comprising 20-30 members, one of which is diagnostic for all nematode-resistant genotypes tested. The presence of non-homologous sequences around the Mi gene may contribute to the suppression of recombination in this region of the genome in crosses heterozygous for Mi. The potential of 'walking' from closely linked markers to Mi is discussed. 相似文献
29.
Helen L. Henry Chhanda Dutta Noreen Cunningham Raymond Blanchard Robert Penny Chilung Tang Gail Marchetto Shu-Yin Chou 《The Journal of steroid biochemistry and molecular biology》1992,41(3-8)
The synthesis of 1,25(OH)2D3 is a critical control point in the regulation of calcium metabolism, and possibly in the growth and differentiation of a number of cell types. This paper reviews our current understanding of the regulation of this process at the cellular and molecular levels, with the emphasis on the mechanisms of feedback control 1,25(OH)2D3 itself, control of parathyroid hormone, the roles of cyclic AMP dependent protein kinase and protein kinase C, and the interaction between the various intracellular regulators of 1,25(OH)2D3 production. 相似文献
30.
Summary Nitrate fertiliser labelled with15N was applied to a field grown crop of winter wheat. Uptake and assimilation of fertiliser nitrate was studied by monitoring the appearance of labelled nitrate and labelled amino acids in the xylem sap. Shortly after applying15N-nitrate to the soil about 30 per cent of recently absorbed15N was in the reduced form, indicating that roots of cereal crops can make a substantial contribution in reducing nitrate. Seasonal changes in crop growth andin vivo NRA are also described. 相似文献