全文获取类型
收费全文 | 618392篇 |
免费 | 67626篇 |
国内免费 | 378篇 |
出版年
2021年 | 5358篇 |
2018年 | 6555篇 |
2017年 | 6342篇 |
2016年 | 8914篇 |
2015年 | 12159篇 |
2014年 | 14320篇 |
2013年 | 19949篇 |
2012年 | 23078篇 |
2011年 | 23288篇 |
2010年 | 15533篇 |
2009年 | 14371篇 |
2008年 | 20252篇 |
2007年 | 20863篇 |
2006年 | 19498篇 |
2005年 | 18796篇 |
2004年 | 18524篇 |
2003年 | 17602篇 |
2002年 | 16943篇 |
2001年 | 29296篇 |
2000年 | 28956篇 |
1999年 | 22952篇 |
1998年 | 7913篇 |
1997年 | 8140篇 |
1996年 | 7479篇 |
1995年 | 6932篇 |
1994年 | 6614篇 |
1993年 | 6657篇 |
1992年 | 17719篇 |
1991年 | 16885篇 |
1990年 | 16286篇 |
1989年 | 15781篇 |
1988年 | 14443篇 |
1987年 | 13444篇 |
1986年 | 12512篇 |
1985年 | 12400篇 |
1984年 | 10317篇 |
1983年 | 8688篇 |
1982年 | 6698篇 |
1981年 | 6002篇 |
1980年 | 5721篇 |
1979年 | 9340篇 |
1978年 | 7275篇 |
1977年 | 6672篇 |
1976年 | 6045篇 |
1975年 | 6559篇 |
1974年 | 7115篇 |
1973年 | 6885篇 |
1972年 | 6217篇 |
1971年 | 5624篇 |
1970年 | 4866篇 |
排序方式: 共有10000条查询结果,搜索用时 15 毫秒
91.
Isolation of viral coat protein mutants with altered assembly and aggregation properties 总被引:1,自引:0,他引:1 下载免费PDF全文
A method was developed to screen bacteria for synthesis of mutant proteins with altered assembly and solubility properties using bacteriophage MS2 coat protein as a model self-associating protein. Colonies expressing coat protein from a plasmid were covered with an agarose overlay under conditions that caused the lysis of some of the cells in each colony. The proteins thus liberated diffused through the overlay at rates depending on their molecular sizes. After transfer of the proteins to a nitrocellulose membrane, probing with coat protein-specific antiserum revealed spots whose sizes and intensities were related to the aggregation state of coat protein. The method was employed in the isolation of assembly defective mutants and to find soluble variants of an aggregation-prone coat protein mutant. 相似文献
92.
HIF-1 expression in healing wounds: HIF-1alpha induction in primary inflammatory cells by TNF-alpha 总被引:12,自引:0,他引:12
93.
As part of systematic studies of the genus Porphyrain New Zealand, constituent sugar analyses of cell wall polysaccharidesin situ in dry thalli were found to yield data that weretaxonomically informative. Variation in constituent sugar levels betweenspecieswas sufficient in some cases to be useful in species differentiation. Thereproductive state of thallus regions had a significant impact on the levels ofconstituent sugars, whereas storage of dried thalli for eight months had noeffect. Three epiphytic taxa currently classified as species ofPorphyra appear to be incorrectly placed within the genus,as their constituent sugars and the levels of these sugars differed markedlyfrom those of all other species examined. 相似文献
94.
Peanut (Arachis hypogaea) agglutinin (PNA) is extensively used as tumour marker as it strongly recognises the cancer specific T antigen (Galβ1→3GalNAc-), but not its sialylated version. However, an additional specificity towards Galβ1→4GlcNAc (LacNAc), which is not tumour specific, had been attributed to PNA. For correct interpretation of lectin histochemical results we examined PNA sugar specificity using naturally occurring or semi-synthetic glycoproteins, matrix-immobilised galactosides and lectin-binding tissue glycoproteins, rather than mono- or disaccharides as ligands. Dot-blots, transfer blots or polystyrene plate coatings of the soluble glycoconjugates were probed with horse-radish peroxidase (HRP) conjugates of PNA and other lectins of known specificity. Modifications of PNA-binding glycoproteins, including selective removal of O-linked oligosaccharides and treatment with glycosidases revealed that Galβ1→4GlcNAc (LacNAc) was ineffective while terminal α-linked galactose (TAG) as well as exposed T antigen (Galβ1→3 GalNAc-) was excellent as sugar moiety in glycoproteins for their recognition by PNA. When immobilised, melibiose was superior to lactose in PNA binding. Results were confirmed using TAG-specific human serum anti-α-galactoside antibody. 相似文献
95.
The cell surface of the parasitic protozoan Leishmania mexicana is coated by glycosylphosphatidylinositol (GPI)-anchored glycoproteins, a GPI-anchored lipophosphoglycan and a class of free GPI glycolipids. To investigate whether the anchor or free GPIs are required for parasite growth we cloned the L.mexicana gene for dolichol-phosphate-mannose synthase (DPMS) and attempted to create DPMS knockout mutants by targeted gene deletion. DPMS catalyzes the formation of dolichol-phosphate mannose, the sugar donor for all mannose additions in the biosynthesis of both the anchor and free GPIs, except for a alpha1-3-linked mannose residue that is added exclusively to the free GPIs and lipophosphoglycan anchor precursors. The requirement for dolichol-phosphate-mannose in other glycosylation pathways in L.mexicana is minimal. Deletion of both alleles of the DPMS gene (lmdpms) consistently resulted in amplification of the lmdpms chromosomal locus unless the promastigotes were first transfected with an episomal copy of lmdpms, indicating that lmdpms, and possibly GPI biosynthesis, is essential for parasite growth. As evidence presented in this and previous studies indicates that neither GPI-anchored glycoproteins nor lipophosphoglycan are required for growth of cultured parasites, it is possible that the abundant and functionally uncharacterized free GPIs are essential membrane components. 相似文献
96.
Rigorous extraction methods coupled with capillary gel electrophoresis (CGE) provide a basis for a nonradiolabel assay for quantitation of intact antisense drug and its numerous chain-shortened metabolites. As part of the validation of the CGE method, we compared the quantitation of unlabeled ISIS 3521 (ISI 641A) and its chain-shortened metabolites with total radioactivity of [(35)S]-ISIS 3521. ISIS 3521 was labeled on the fifth nucleotide linkage from the 5'-end with (35)S by well-established methods. Multiple tissues collected from rats after administration of [(35)S]-ISIS 3521 were assayed by both radiolabel (liquid scintillation spectroscopy) and CGE methods. The CGE method provided accurate quantitation of the drug and its metabolites in kidney cortex and liver tissues. The correlation between methods for multiple tissues over time was excellent with 88.5% of the measurements being statistically equivalent. These data suggest that CGE is an accurate means of quantitating oligonucleotide in tissue and that it compares favorably with traditional radiochemical techniques. Clearance half-lives for total measurable oligonucleotides were equivalent to clearance of total radioactivity in both liver and kidney with the longest clearance half-life associated with the kidney. 相似文献
97.
98.
99.
Wei CHEN Liqing PENG Lichun JIANG David A.PIKE Christopher R.FRIESEN Gregory BROWN 《亚洲两栖爬行动物研究(英文版)》2018,(1)
Environmental unpredictability can influence strategies of maternal investment among eggs within a clutch. Models predict that breeding females should adopt a diversified bet-hedging strategy in unpredictable environments, but empirical field evidence from Asia is scarce. Here we tested this hypothesis by exploring spatial patterns in egg size along an altitudinal gradient in a frog species(Rana kukunoris) inhabiting the Tibetan Plateau. Within-clutch variability in egg size increased as the environment became variable(e.g., lower mean monthly temperature and mean monthly rainfall at higher altitudes), and populations in environments with more unpredictable rainfall produced eggs that were smaller and more variable in size. We provide support for a diversified bet-hedging strategy in high-altitude environments, which experience dynamic weather patterns and therefore are of unpredictable environmental quality. This strategy may be an adaptive response to lower environmental quality and higher unpredictable environmental variance. Such a strategy should increase the likelihood of breeding success and maximize maternal lifetime fitness by producing offspring that are adapted to current environmental conditions. We speculate that in high-altitude environments prone to physical disturbance, breeding females are unable to consistently produce the optimal egg size due to physiological constraints imposed by environmental conditions(e.g., duration of the active season, food availability). Species and populations whose breeding strategies are adapted to cope with uncertain environmental conditions by adjusting offspring size and therefore quality show a remarkable degree of ability to cope with future climatic changes. 相似文献
100.
A A Gureeva A V Sanin V O Rybin N S Barteneva I A Lapaeva 《Zhurnal mikrobiologii, epidemiologii, i immunobiologii》1986,(5):63-66
Effect of B. pertussis lymphocytosis-promoting factor (LPF) on the lympho-hematopoietic system of mice was studied. The injection of LPF was shown to sharply enhance endogenous colony formation and to induce a severe depletion of thymus cells, reaching its maximum of day 4. Thymocytes obtained on day 2 or 3 after the injection of LPF produced a suppressive effect on endogenous colony formation. The proliferative activity of hematopoietic stem cells sharply increased under the influence of LPF, though it had no radioprotective action. On the following day after the injection of LPF a steep rise in the number of hematopoietic stem cells was observed in the blood of mice: their content increased 20-fold in comparison with the control level. These data may be important for the evaluation of the side effects of pertussis vaccine on the lympho-hematopoietic system. 相似文献