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1.
The EF‐1α promoter maintains high‐level transgene expression from episomal vectors in transfected CHO‐K1 cells 下载免费PDF全文
Xi Zhang Danhua Xu Qin Li Junhe Zhang Tianyun Wang 《Journal of cellular and molecular medicine》2017,21(11):3044-3054
In our previous study, we demonstrated that episomal vectors based on the characteristic sequence of matrix attachment regions (MARs) and containing the cytomegalovirus (CMV) promoter allow transgenes to be maintained episomally in Chinese hamster ovary (CHO) cells. However, the transgene expression was unstable and the number of copies was low. In this study, we focused on enhancers, various promoters and promoter variants that could improve the transgene expression stability, expression magnitude (level) and the copy number of a MAR‐based episomal vector in CHO‐K1 cells. In comparison with the CMV promoter, the eukaryotic translation elongation factor 1 α (EF‐1α, gene symbol EEF1A1) promoter increased the transfection efficiency, the transgene expression, the proportion of expression‐positive clones and the copy number of the episomal vector in long‐term culture. By contrast, no significant positive effects were observed with an enhancer, CMV promoter variants or CAG promoter in the episomal vector in long‐term culture. Moreover, the high‐expression clones harbouring the EF‐1α promoter tended to be more stable in long‐term culture, even in the absence of selection pressure. According to these findings, we concluded that the EF‐1α promoter is a potent regulatory sequence for episomal vectors because it maintains high transgene expression, transgene stability and copy number. These results provide valuable information on improvement of transgene stability and the copy number of episomal vectors. 相似文献
2.
《DNA Repair》2017
Infliction of DNA damage initiates a complex cellular reaction – the DNA damage response – that involves both signaling and DNA repair networks with many redundancies and parallel pathways. Here, we reveal the three strategies that the simple multicellular eukaryote, C. elegans, uses to deal with DNA damage induced by light. Separately inactivating repair or replicative bypass of photo-lesions results in cellular hypersensitivity towards UV-light, but impeding repair of replication associated DNA breaks does not. Yet, we observe an unprecedented synergistic relationship when these pathways are inactivated in combination. C. elegans mutants that lack nucleotide excision repair (NER), translesion synthesis (TLS) and alternative end joining (altEJ) grow undisturbed in the dark, but become sterile when grown in light. Even exposure to very low levels of normal daylight impedes animal growth. We show that NER and TLS operate to suppress the formation of lethal DNA breaks that require polymerase theta-mediated end joining (TMEJ) for their repair. Our data testifies to the enormous genotoxicity of light and to the demand of multiple layers of protection against an environmental threat that is so common. 相似文献
3.
In this article, we discuss molecular mechanisms involved in the evolution of amygdala kindling and the episodic loss of response
to pharmacological treatments during tolerance development. These phenomena allow us to consider how similar principles (in
different neurochemical systems) could account for illness progression, cyclicity, and drug tolerance in affective disorders.
We describe the phenomenon of amygdala-kindled seizures episodically breaking through effective daily pharmacotherapy with
carbamazepine and valproate, suggesting that these observations could reflect the balance of pathological vs compensatory
illness-induced changes in gene expression. Under certain circumstances, amygdala-kindled animals that were initially drug
responsive can develop highly individualized patterns of seizure breakthroughs progressing toward a complete loss of drug
efficacy. This initial drug efficacy may reflect the combination of drug-related exogenous neurochemical mechanisms and illness-induced
endogenous compensatory mechanisms. However, we postulate that when seizures are inhibited, the endogenous illness-induced
adaptations dissipate (the “time-off seizure” effect), leading to the re-emergence of seizures, a re-induction of a new, but
diminished, set of endogenous compensatory mechanisms, and a temporary period of renewed drug efficacy. As this pattern repeats,
an intermittent or cyclic response to the anticonvulsant treatment emerges, leading toward complete drug tolerance.
We also postulate that the cyclic pattern accelerates over time because of both the failure of robust illness-induced endogenous
adaptations to emerge and the progression in pathophysiological mechanisms (mediated by long-lasting changes in gene expression
and their downstream consequences) as a result of repeated occurrences of seizures. In this seizure model, this pattern can
be inhibited and drug responsivity can be temporarily reinstated by several manipulations, including lowering illness drive
(decreasing the stimulation current.), increasing drug dosage, switching to a new drug that does not show crosstolerance to
the original medication, or temporarily discontinuing treatment, allowing the illness to re-emerge in an unmedicated animal.
Each of these variables is discussed in relation to the potential relevance to the emergence, progression, and suppression
of individual patterns of episodic cyclicity in the recurrent affective disorders. A variety of clinical studies are outlined
that specifically test the hypotheses derived from this formulation. Data from animal studies suggest that illness cyclicity
can develop from the relative ratio between primary pathological processes and secondary endogenous adaptations (assisted
by exogenous medications). If this proposition is verified, it further suggests that illness cyclicity is inherent to the
neurobiological processes of episode emergence and amelioration, and one does not need to postulate a separate defect in the
biological clock. The formulation predicts that early and aggressive long-term interventions may be optimal in order to prevent
illness emergence and progression and its associated accumulating neurobiological, vulnerability factors. 相似文献
4.
Learning-Induced Expression of Meningeal Ependymin mRNA and Demonstration of Ependymin in Neurons and Glial Cells 总被引:1,自引:0,他引:1
Stefan Rother Rupert Schmidt Wolfgang Brysch †Karl-Hermann Schlingensiepen 《Journal of neurochemistry》1995,65(4):1456-1464
Abstract: The turnover of a CNS-specific cell adhesion glycoprotein, ependymin, has earlier been found to increase during periods of neuronal plasticity. Here, ependymin mRNA expression was analyzed by semiquantitative in situ hybridization in goldfish. Learning of an active avoidance response resulted in a significant increase in ependymin mRNA expression 20 min to 4 h after acquisition of the task. In contrast, yoked control animals that were exposed to the same numbers of conditioned and unconditioned stimuli in a random, unpaired manner exhibited a strong down-regulation of ependymin mRNA. Hybridization signals were also increased by injection of anti-ependymin antiserum into brain ventricles. Ependymin mRNA was exclusively localized to reticular-shaped fibroblasts of the inner endomeningeal cell layer. Immunoelectron microscopic investigation, however, revealed ependymin also in distinct neuronal and glial cell populations in which no ependymin mRNA had been detected. Uptake of meningeal protein factors into glial and neuronal cells may therefore be of functional importance for plastic adaptations of the CNS. 相似文献
5.
6.
Solar Cells: Role of Microstructure in Oxygen Induced Photodegradation of Methylammonium Lead Triiodide Perovskite Films (Adv. Energy Mater. 20/2017) 下载免费PDF全文
7.
8.
Streptococcus pyogenes is commonly found on pharynx, mouth and rarely on skin, lower gastrointestinal tract. It is a potential pathogen causing tonsillitis, pneumonia, endocarditis. The present study was undertaken to study the effects of low shear modeled microgravity on growth, morphology, antibiotic resistance, cross-stress resistance to various stresses and alteration in gene expression of S. pyogenes. The growth analysis performed using UV–Visible spectroscopy indicated decrease in growth of S. pyogenes under low shear modeled microgravity. Morphological analysis by Bio-transmission electron microscopy (TEM), Bio-scanning electron microscopy (SEM) did not reveal much difference between normal and low shear modeled microgravity grown S. pyogenes. The sensitivity of S. pyogenes to antibiotics ampicillin, penicillin, streptomycin, kanamycin, hygromycin, rifampicin indicates that the bacterium is resistant to hygromycin. Further S. pyogenes cultured under low shear modeled microgravity was found to be more sensitive to ampicillin and rifampicin as compared with normal gravity grown S. pyogenes. The bacteria were tested for the acid, osmotic, temperature and oxidative cross stress resistances. The gene expression of S. pyogenes under low shear modeled microgravity analyzed by microarray revealed upregulation of 26 genes and down regulation of 22 genes by a fold change of 1.5. 相似文献
9.
Sally A. Amundson Michael Bittner Paul Meltzer Jeffrey Trent Albert J. Fornace Jr. 《Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology》2001,129(4)
The responses to ionizing radiation and other genotoxic environmental stresses are complex and are regulated by a number of overlapping molecular pathways. One such stress signaling pathway involves p53, which regulates the expression of over 100 genes already identified. It is also becoming increasingly apparent that the pattern of stress gene expression has some cell type specificity. It may be possible to exploit these differences in stress gene responsiveness as molecular markers through the use of a combined informatics and functional genomics approach. The techniques of microarray analysis potentially offer the opportunity to monitor changes in gene expression across the entire set of expressed genes in a cell or organism. As an initial step in the development of a functional genomics approach to stress gene analysis, we have recently demonstrated the utility of cDNA microarray hybridization to measure radiation-stress gene responses and identified a number of previously unknown radiation-regulated genes. The responses of some of these genes to DNA-damaging agents vary widely in cell lines from different tissues of origin and different genetic backgrounds. While this again highlights the importance of a cellular context to genotoxic stress responses, it also raises the prospect of expression-profiling of cell lines, tissues, and tumors. Such profiles may have a predictive value if they can define regions of ‘expression space’ that correlate with important endpoints, such as response to cancer therapy regimens, or identification of exposures to environmental toxins. 相似文献
10.
Root tensile strength and root distribution of typical Mediterranean plant species and their contribution to soil shear strength 总被引:13,自引:0,他引:13
S. De Baets J. Poesen B. Reubens K. Wemans J. De Baerdemaeker B. Muys 《Plant and Soil》2008,305(1-2):207-226
In Mediterranean environments, gully erosion is responsible for large soil losses. It has since long been recognized that
slopes under vegetation are much more resistant to soil erosion processes compared to bare soils and improve slope stability.
Planting or preserving vegetation in areas vulnerable to erosion is therefore considered to be a very effective soil erosion
control measure. Re-vegetation strategies for erosion control rely in most cases on the effects of the above-ground biomass
in reducing water erosion rates, whereas the role of the below-ground biomass is often neglected or underestimated. While
the above-ground biomass can temporally disappear in semi-arid environments, roots may still be present underground and play
an important role in protecting the topsoil from being eroded. In order to evaluate the potential of plant species growing
in Mediterranean environments to prevent shallow mass movements on gully or terrace walls, the root reinforcement effect of
25 typical Mediterranean matorral species (i.e. shrubs, grasses herbs, small trees) was assessed, using the simple perpendicular
model of Wu et al. (Can Geotech J 16:19–33, 1979). As little information is available on Mediterranean plant root characteristics,
root distribution data were collected in SE-Spain and root tensile strength tests were conducted in the laboratory. The power
root tensile strength–root diameter relationships depend on plant species. The results show that the shrubs Salsola genistoides Juss. Ex Poir. and Atriplex halimus L. have the strongest roots, followed by the grass Brachypodium retusum (Pers.) Beauv. The shrubs Nerium oleander L. and the grass Avenula bromoides (Gouan) H. Scholz have the weakest roots in tension. Root area ratio for the 0–0.1 m topsoil ranges from 0.08% for the grass
Piptatherum miliaceum (L.) Coss to 0.8% for the tree Tamarix canariensis Willd. The rush Juncus acutus L. provides the maximum soil reinforcement to the topsoil by its roots (i.e. 304 kPa). Grasses also increase soil shear strength
significantly (up to 244 kPa in the 0–0.1 m topsoil for Brachypodium retusum (Pers.) Beauv.). The shrubs Retama sphaerocarpa (L.) Boiss. and Anthyllis cytisoides L. are increasing soil shear strength to a large extent as well (up to 134 and 160 kPa respectively in the 0–0.10 m topsoil).
Whereas grasses and the rush Juncus acutus L. increase soil shear strength in the topsoil (0–0.10 m) to a large extent, the shrubs Anthyllis cytisoides (L.), Retama sphaerocarpa (L.) Boiss., Salsola genistoides Juss. Ex Poir. and Atriplex halimus L. strongly reinforce the soil to a greater depth (0–0.5 m). As other studies reported that Wu’s model overestimates root
cohesion values, reported root cohesion values in this study are maximum values. Nevertheless, the calculated cohesion values
are used to rank species according to their potential to reinforce the soil. 相似文献