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Majumdar R  Railkar R  Dighe RR 《Proteins》2011,79(11):3108-3122
Single chain fragment variables (ScFvs) have been extensively employed in studying the protein-protein interactions. ScFvs derived from phage display libraries have an additional advantage of being generated against a native antigen, circumventing loss of information on conformational epitopes. In the present study, an attempt has been made to elucidate human chorionic gonadotropin (hCG)-luteinizing hormone (LH) receptor interactions by using a neutral and two inhibitory ScFvs against hCG. The objective was to dock a computationally derived model of these ScFvs onto the crystal structure of hCG and understand the differential roles of the mapped epitopes in hCG-LH receptor interactions. An anti-hCG ScFv, whose epitope was mapped previously using biochemical tools, served as the positive control for assessing the quality of docking analysis. To evaluate the role of specific side chains at the hCG-ScFv interface, binding free energy as well as residue interaction energies of complexes in solution were calculated using molecular mechanics Poisson-Boltzmann/surface area method after performing the molecular dynamic simulations on the selected hCG-ScFv models and validated using biochemical and SPR analysis. The robustness of these calculations was demonstrated by comparing the theoretically determined binding energies with the experimentally obtained kinetic parameters for hCG-ScFv complexes. Superimposition of hCG-ScFv model onto a model of hCG complexed with the 51-266 residues of LH receptor revealed importance of the residues previously thought to be unimportant for hormone binding and response. This analysis provides an alternate tool for understanding the structure-function analysis of ligand-receptor interactions.  相似文献   
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Abstract Developmental curves for the sister species Chrysomya chloropyga (Wiedemann, 1818) and Chrysomya putoria (Wiedemann, 1830) (Diptera: Calliphoridae) were established at eight and 10 different constant temperatures, respectively, using developmental landmarks and body length as measures of age. The thermal summation constants (K) and developmental threshold (D0) were calculated for five developmental landmarks using a previously described method. Isomorphen and isomegalen diagrams were also constructed for the purpose of estimating postmortem intervals (PMIs). Chrysomya chloropyga had an average developmental threshold value (D0) of 10.91 °C (standard error [SE] = 0.94 °C, n = 5), significantly lower than that of C. putoria (13.42 °C, SE = 0.45 °C, n = 5) (paired t‐test: t = ? 4.63, d.f. = 8, P < 0.00). Similarly, K values for C. chloropyga were larger than those for C. putoria for all developmental events except onset of the wandering phase. These are the first data that can be used to calculate minimum PMIs and predict population growth of C. chloropyga and C. putoria in Africa; the data indicate that developmental data for one of these species cannot be used as surrogate data for the sister species.  相似文献   
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Many herbivorous insects sequester defensive chemicals from their host plants. We tested sequestration fitness costs in the specialist moth Utetheisa ornatrix (Lepidoptera: Arctiidae). We added pyrrolizidine alkaloids (PAs) to an artificial diet at different concentrations. Of all the larval and adult fitness components measured, only development time was negatively affected by PA concentration. These results were repeated under stressful laboratory conditions. On the other hand, the amount of PAs sequestered greatly increased with the diet PA concentration. Absence of a detectable negative effect does not necessarily imply a lack of costs if all individuals express the biochemical machinery of detoxification and sequestration constitutively. Therefore, we used qPCR to show that expression of the gene used to detoxify PAs, pyrrolizidine‐alkaloid‐N‐oxygenase (pno), increased 41‐fold in our highest PA treatment. Nevertheless, fitness components were affected only slightly or not at all, suggesting that sequestration in this species does not incur a strong cost. The apparent lack of costs has important implications for our understanding of the evolution of ecological interactions; for example, it implies that selection by specialist herbivores may decrease the levels of certain chemical defences in plant populations.  相似文献   
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Turnbull LA  Rees M  Purves DW 《Ecology letters》2008,11(10):1037-1046
Equalising trade-offs, such as seed mass vs. number, have been invoked to reconcile neutral theory with observed differences between species. This is an appealing explanation for the dramatic seed size variation seen within guilds of otherwise similar plants: under size-symmetric competition, where resource capture is proportional to mass, the outcome of competition should be insensitive to whether species produce many small seeds or few large ones. However, under this assumption, stochastic variation in seed rain leads to exclusion of all but the smallest-seeded species. Thus stochasticity in seed arrivals, a process that was previously thought to generate drift, instead results in deterministic competitive exclusion. A neutral outcome is possible under one special case of a more general equalising framework, where seed mass affects survival but not competition. Further exploration of the feasibility of neutral trade-offs is needed to understand the respective roles of neutrality and niche structure in community dynamics.  相似文献   
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Grazer control of periphyton biomass has been addressed in numerous experimental studies in all kinds of aquatic habitats. In this meta‐analysis, the results of 865 experiments are quantitatively synthesized in order to address the following questions: (i) Do lotic, lentic, and marine ecosystems differ in their degree of grazer control of periphyton biomass? (ii) Which environmental variables affect the degree of grazer control? (iii) How much does the result of these experiments depend on facets of experimental design? Across all ecosystems, the grazers removed on average 59% of the periphyton biomass, with grazing being significantly stronger for laboratory (65%) than for field (56%) experiments. Neither field nor lab experiments showed a significant difference among lotic, lentic, and coastal habitats. Among different taxonomic consumer groups, crustaceans (amphipods and isopods) and trichopteran larvae removed the highest proportion of periphyton biomass. Grazer effects increased with increasing algal biomass, with decreasing resource availability and with increasing temperature, especially in field experiments. Grazer effects also increased with increasing total grazer biomass in field experiments but showed the opposite trend in lab experiments, indicating a tendency toward overcrowded lab experiments. Other aspects of experimental design, such as cage type, size, and duration of the study, strongly affected the outcome of the experiments, suggesting that much care has to be placed on the choice of experimental design.  相似文献   
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Antennal proteins of the male fire ant (Solenopsis invicta) were analyzed by two-dimensional gel electrophoresis, with the objective of identifying pheromone-binding proteins, which have not previously been found in ant antennae. The major low-molecular weight protein found in the male fire ant antenna was subjected to Edman degradation to determine the N-terminal amino acid sequence. Degenerate PCR primers based on this sequence were used to obtain a cDNA sequence corresponding to the full-length protein sequence. In-gel trypsin digestion followed by MALDI-TOF mass spectrometry and HPLC-ESI/MS/MS demonstrated that the protein gel spot contained only the protein corresponding to the cDNA sequence obtained by PCR. The sequence is similar to apolipophorin-III, an exchangeable lipid-binding protein. Fire ant apolipophorin-III is expressed in the antenna as well as the head, thorax and abdomen.  相似文献   
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Intergenerational fitness effects on offspring due to the early life of the parent are well studied from the standpoint of the maternal environment, but intergenerational effects owing to the paternal early life environment are often overlooked. Nonetheless, recent laboratory studies in mammals and ecologically relevant studies in invertebrates predict that paternal effects can have a major impact on the offspring's phenotype. These nongenetic, environment‐dependent paternal effects provide a mechanism for fathers to transmit environmental information to their offspring and could allow rapid adaptation. We used the bank vole Myodes glareolus, a wild rodent species with no paternal care, to test the hypothesis that a high population density environment in the early life of fathers can affect traits associated with offspring fitness. We show that the protein content in the diet and/or social environment experienced during the father's early life (prenatal and weaning) influence the phenotype and survival of his offspring and may indicate adaptation to density‐dependent costs. Furthermore, we show that experiencing multiple environmental factors during the paternal early life can lead to a different outcome on the offspring phenotype than stimulated by experience of a single environmental factor, highlighting the need to study developmental experiences in tandem rather than independent of each other.  相似文献   
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