全文获取类型
收费全文 | 1465篇 |
免费 | 76篇 |
出版年
2022年 | 6篇 |
2021年 | 8篇 |
2018年 | 19篇 |
2017年 | 10篇 |
2016年 | 18篇 |
2015年 | 41篇 |
2014年 | 37篇 |
2013年 | 97篇 |
2012年 | 62篇 |
2011年 | 56篇 |
2010年 | 39篇 |
2009年 | 45篇 |
2008年 | 62篇 |
2007年 | 83篇 |
2006年 | 77篇 |
2005年 | 64篇 |
2004年 | 88篇 |
2003年 | 92篇 |
2002年 | 77篇 |
2001年 | 59篇 |
2000年 | 66篇 |
1999年 | 36篇 |
1998年 | 23篇 |
1997年 | 14篇 |
1996年 | 12篇 |
1995年 | 7篇 |
1994年 | 11篇 |
1993年 | 11篇 |
1992年 | 31篇 |
1991年 | 29篇 |
1990年 | 16篇 |
1989年 | 22篇 |
1988年 | 16篇 |
1987年 | 18篇 |
1986年 | 15篇 |
1985年 | 16篇 |
1984年 | 10篇 |
1983年 | 12篇 |
1982年 | 11篇 |
1981年 | 10篇 |
1980年 | 10篇 |
1979年 | 8篇 |
1978年 | 9篇 |
1977年 | 10篇 |
1975年 | 11篇 |
1974年 | 6篇 |
1973年 | 5篇 |
1970年 | 12篇 |
1969年 | 8篇 |
1967年 | 7篇 |
排序方式: 共有1541条查询结果,搜索用时 453 毫秒
1.
2.
3.
Induction of competence and progression signals in human T lymphocytes by phorbol esters and calcium ionophores 总被引:1,自引:0,他引:1
We have investigated the induction of competence (IL-2 responsiveness) and progression in human T lymphocyte proliferation triggered by phorbol ester and calcium ionophore. The degree of proliferation induced with the phorbol ester, phorbol 12,13-dibutyrate (PDB) and the calcium ionophore ionomycin was dependent on the duration of exposure to these agents, with more than 6 h required for obtaining maximum proliferation. Following brief exposure to both agents for 30 min, which did not cause significant proliferation, T cells became competent to proliferate in response to exogenous interleukin 2 (IL-2). These competent T cells also progressed to DNA synthesis following incubation with PDB in the absence of ionomycin. Induction of competence to proliferate in response to either PDB or IL-2 was blocked by EGTA, suggesting that transmembrane Ca2+ flux was obligatory at this stage. Since other phorbol esters and synthetic diacylglycerols also stimulated DNA synthesis in competent cells, it is likely that progression was triggered by activation of protein kinase C. Following a brief exposure to PDB and ionomycin, subsequent incubation with PDB induced gene expression and secretion of IL-2 and augmented the expression of IL-2 receptors in the competent cells. Thus, we have demonstrated that Ca2+ mobilization is required for rendering T cells competent to express functional IL-2 receptors, to produce IL-2 in response to subsequent incubation with PDB, and that sustained activation of protein kinase C seems necessary for IL-2 production and subsequent progression of competent T cells to DNA synthesis. 相似文献
4.
Norio Masui Yumie Takagi Tetsu Nishikawa Makoto Yanabe Masato Nose Katsunori Sato 《Experimental Animals》2002,51(5):501-503
A new polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis was developed for genetic typing of the mouse Tnfsf6gld mutation. An artificial restriction site was introduced to the mouse Tnfsf6gld mutation by PCR amplification using a modified primer. The three genotypes of the Tnfsf6 locus (Tnfsf6gld/Tnfsf6gld, Tnfsf6gld/+, and +Tnfsf6-gld/+Tnfsf6-gld) could be distinguished clearly and easily. This PCR-RFLP analysis was found to be useful for the identification of the mouse Tnfsf6gld mutation. 相似文献
5.
In poly(A)+RNA extracted from a lactating goat mammary gland, mRNA of about 750 nucleotides was shown to encode pre alpha-lactalbumin by using in vitro translation and immunoprecipitation. From the total poly(A)+RNA, the cDNA library was constructed using the Escherichia coli plasmid pUC18; it was screened with the oligodeoxyribonucleotide probe corresponding to the amino acid sequence of Trp60-Gln65 of goat alpha-lactalbumin. A plasmid containing almost full-length cDNA of goat pre alpha-lactalbumin, pGLA-1, was identified. The cDNA insert of pGLA-1 comprises 727 base pairs and contains the signal peptide and mature protein sequence. 相似文献
6.
7.
Expression of goat alpha-lactalbumin in Escherichia coli and its refolding to biologically active protein 总被引:1,自引:0,他引:1
A cDNA encoding the mature region of goat alpha-lactalbumin and the 3'-non-coding region was fused to cDNA of the N-terminal half of porcine adenylate kinase which had been placed under the control of the tac promoter in an expression vector in Escherichia coli. In addition, a methionine codon was inserted between the two cDNAs. When the plasmid carried the full-length 3'-non-coding region, little accumulation of the fused protein was observed. However, the deletion of two-thirds of the 3'-non-coding region produced significant expression of the fused protein in E. coli strain JM105. Since goat alpha-lactalbumin contains no methionine residue, the mature goat alpha-lactalbumin was isolated by CNBr digestion of the fused insoluble protein and refolded using thioredoxin. The homogeneous and biologically active goat alpha-lactalbumin was purified by Ca2+ ion-dependent hydrophobic chromatography. 相似文献
8.
Determination of base composition of DNA by high performance liquid chromatography of its nuclease P1 hydrolysate 总被引:1,自引:0,他引:1
The base composition of DNA was directly determined by high performance liquid chromatography (HPLC) of its nuclease P1 hydrolysate. This method can satisfactorily be employed even if the sample of DNA contains RNA or the amount of the sample is very small. 相似文献
9.
Prerequisite for the induction of lymphokine-activated killer cells from T lymphocytes 总被引:5,自引:0,他引:5
H Sawada T Abo S Sugawara K Kumagai 《Journal of immunology (Baltimore, Md. : 1950)》1988,140(10):3668-3673
Human blood mononuclear cells were separated into Leu-11+7-NK, Leu-11-7+, and Leu-11-7-T cells by means of a combination of the Percoll gradient method and C-mediated cytolysis using mAb. When purified Leu-11+7-NK, Leu-11-7+, and Leu-11-7-T cells were cultured with rIL 2 (500 U/ml) for 6 days in a medium supplemented with 10% FCS, Leu-11+7-NK cells responded at the maximum level and Leu-11-7+ cells responded moderately as shown by both cell-proliferation response and cytotoxic activity generated. On the other hand, Leu-11-7-T cells did not respond at all to rIL-2. However, when Leu-11-7-T cells were cultured with rIL-2 in a medium supplemented with 10% autologous serum, they showed considerable responsiveness to rIL-2. In addition, much greater response to Leu-11-7-T cells were produced by the addition of monocytes. Monocyte cytokines, neither IL 1, IFN-gamma, TNF, nor their combination were able to substitute for monocytes in the induction culture. In contrast, the response level of Leu-11+7- NK cells remained unchanged irrespective of supplementation with autologous serum to medium or the addition of monocytes to the culture. These results indicated that culture conditions in the experiments significantly affected the results as to determination of lymphokine-activated killer cell precursors, especially the result pertaining to the conversion of T lymphocytes to lymphokine-activated killer cells. Under appropriate conditions, not only NK cells but also T cells are important precursors of lymphokine-activated killer cells. 相似文献
10.
Tetsu Hirata 《Ecological Research》1992,7(1):31-42
Successional changes of sessile organisms over about 3 years on concrete plates immersed at depths of 1.0, 2.5, 4.0 and 5.5
m in Nabeta Bay, Shimoda, Pacific coast of Japan, were investigated from their species composition, percentage cover, and
the structure, metabolism, biomass and maturity of the community for the purpose of verifying the hypothesis of succession
by Margalef (1968) and Odum (1969). The process of succession was divided into five stages from a cluster analysis based on
Horn's (1966) similarity in community structure. The five stages were characterized from dominant, species and 18 community
attributes in terms of structure, metabolism, biomass and maturity. With the advance of stages, two large-sized, long-lived
species, the kelpEisenia bicyclis and the oysterCrassostrea nippona, dominated; 18 community attributes showed a directional succession ending in a constant community structure, low community
metabolism, accumulation of community biomass except chlorophylla and an enhanced degree of community maturity. This directionality in succession of community attributes was valid for the
prediction of Margalef and Odum, despite a tendency for estimated daily energy budget to be more heterotrophic in such an
open system. From these results, it was suggested that the definition of succession by Odum (1969) should be interpreted as
the regularity in shift of dominant species, not individual species, and as the control of system by feedback mechanism developing
with succession rather than community-controlled mechanism. 相似文献