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11.
Targeting of T7 RNA polymerase to tobacco nuclei mediated by an SV40 nuclear location signal 总被引:9,自引:0,他引:9
Michael W. Lassner Aubrey Jones Steve Daubert Luca Comai 《Plant molecular biology》1991,17(2):229-234
We have expressed two T7 RNA polymerase genes by electroporation into tobacco protoplasts. One of the genes was modified by inserting nucleotides encoding a viral nuclear localization signal (NLS) from the large T antigen of SV40. Both T7 RNA polymerase genes directed synthesis of a ca. 100 kDa protein in the electroporated protoplasts. T7 RNA polymerase activity was detected in extracts of protoplasts electroporated with both genes. Immunofluorescence analysis of these protoplasts indicated that only the polymerase carrying the NLS accumulated in the cell nucleus. These experiments suggest that mechanisms involved in the transport from the cytoplasm to the nucleus are similar in plant and animal cells. This system demonstrates the feasibility of T7 RNA polymerase-based approaches for the high-level expression of introduced genes in plant cells. 相似文献
12.
《Bioscience, biotechnology, and biochemistry》2013,77(7):1372-1377
The pig is an important animal for both agricultural and medical purposes. However, the number of pig-derived cell lines is relatively limited when compared with mouse- and human-derived lines. We established in this study a retroviral conditional expression system for the Simian vacuolating virus 40 large T fragment (SV40T) which allowed us to efficiently establish pig embryonic fibroblast cell lines. The established cell lines showed high levels of cell proliferation and resistance to cellular senescence. A chromosome analysis showed that 84% of the cells had the normal karyotype. Transient expression of the Cre recombinase allowed us to excise the SV40T fragment from the genome. The development of this research tool will enable us to quickly establish new cell lines derived from various animals. 相似文献
13.
Triplication of a unique genetic segment in a simian virus 40-like virus of human origin and evolution of new viral genomes 总被引:4,自引:0,他引:4
The non-defective (heavy) virions from a simian virus 40-like virus (DAR virus) isolated from human brain have been serially passaged at high input multi-plicities in primary monkey kidney cells. The 32P-labeled, progeny DAR-viral genomes have been purified and tested for sensitivity to the RI restriction endouclease from Escherichia coli (Eco RI3 restriction nuclease). The parental DAR-viral genomes share many physical properties with “standard” simian virus 40 DNA and are cleaved once by the Eco RI restriction nuclease. After the fourth serial passage, three populations of genomes could be distinguished: Eco RI resistant, Eco RI sensitive (one cleavage site) and Eco RI “supersensitive” (three, symmetrically-located, cleavage sites). The Eco RI cleavage product of the “supersensitive” form is one-third the physical size (10.4 S) of simian virus 40 DNA and reassociates about three times more rapidly than sheared, denatured simian virus 40 DNA. From the fourth to the eighth serial passages, the genomes containing this specific triplication of viral DNA sequences were selected for and became the predominant viral DNA species. 相似文献
14.
I. L. Sun W. Toole-Simms F. L. Crane D. J. Morré H. Löw J. Y. Chou 《Journal of bioenergetics and biomembranes》1988,20(3):383-391
Retinoic acid inhibits the reduction of diferric transferrin through the transplasma membrane electron transport system on fetal rat liver cells infected with a temperature-sensitive SV40 virus when the cells are in the nontransformed state cultured at 40°C. When the cells are in the transformed state (grown at the permissive 33°C temperature), retinoic acid does not inhibit the diferric transferrin reduction. Inhibition of activity of nontransformed cells is specific for retinoic acid with only slight inhibition by retinol and retinyl acetate at higher concentrations. Isolated rat liver plasma membrane NADH diferric transferrin reductase is also inhibited by retinoic acid. The effect of transformation with SV40 virus to decrease susceptibility to retinoic acid inhibition stands in contrast to much greater adriamycin inhibition of diferric transferrin reduction in the transformed cells than in nontransformed cells. 相似文献
15.
Heat shock protein (Hsp)40s play an essential role in protein metabolism by regulating the polypeptide binding and release cycle of Hsp70. The Hsp40 family is large, and specialized family members direct Hsp70 to perform highly specific tasks. Type I and Type II Hsp40s, such as yeast Ydj1 and Sis1, are homodimers that dictate functions of cytosolic Hsp70, but how they do so is unclear. Type I Hsp40s contain a conserved, centrally located cysteine-rich domain that is replaced by a glycine- and methionine-rich region in Type II Hsp40s, but the mechanism by which these unique domains influence Hsp40 structure and function is unknown. This is the case because high-resolution structures of full-length forms of these Hsp40s have not been solved. To fill this void, we built low-resolution models of the quaternary structure of Ydj1 and Sis1 with information obtained from biophysical measurements of protein shape, small-angle X-ray scattering, and ab initio protein modeling. Low-resolution models were also calculated for the chimeric Hsp40s YSY and SYS, in which the central domains of Ydj1 and Sis1 were exchanged. Similar to their human homologs, Ydj1 and Sis1 each has a unique shape with major structural differences apparently being the orientation of the J domains relative to the long axis of the dimers. Central domain swapping in YSY and SYS correlates with the switched ability of YSY and SYS to perform unique functions of Sis1 and Ydj1, respectively. Models for the mechanism by which the conserved cysteine-rich domain and glycine- and methionine-rich region confer structural and functional specificity to Type I and Type II Hsp40s are discussed. 相似文献
16.
Fleishman SJ Whitehead TA Strauch EM Corn JE Qin S Zhou HX Mitchell JC Demerdash ON Takeda-Shitaka M Terashi G Moal IH Li X Bates PA Zacharias M Park H Ko JS Lee H Seok C Bourquard T Bernauer J Poupon A Azé J Soner S Ovali SK Ozbek P Tal NB Haliloglu T Hwang H Vreven T Pierce BG Weng Z Pérez-Cano L Pons C Fernández-Recio J Jiang F Yang F Gong X Cao L Xu X Liu B Wang P Li C Wang C Robert CH Guharoy M Liu S Huang Y Li L Guo D Chen Y Xiao Y London N Itzhaki Z Schueler-Furman O Inbar Y Potapov V 《Journal of molecular biology》2011,414(2):289-302
The CAPRI (Critical Assessment of Predicted Interactions) and CASP (Critical Assessment of protein Structure Prediction) experiments have demonstrated the power of community-wide tests of methodology in assessing the current state of the art and spurring progress in the very challenging areas of protein docking and structure prediction. We sought to bring the power of community-wide experiments to bear on a very challenging protein design problem that provides a complementary but equally fundamental test of current understanding of protein-binding thermodynamics. We have generated a number of designed protein-protein interfaces with very favorable computed binding energies but which do not appear to be formed in experiments, suggesting that there may be important physical chemistry missing in the energy calculations. A total of 28 research groups took up the challenge of determining what is missing: we provided structures of 87 designed complexes and 120 naturally occurring complexes and asked participants to identify energetic contributions and/or structural features that distinguish between the two sets. The community found that electrostatics and solvation terms partially distinguish the designs from the natural complexes, largely due to the nonpolar character of the designed interactions. Beyond this polarity difference, the community found that the designed binding surfaces were, on average, structurally less embedded in the designed monomers, suggesting that backbone conformational rigidity at the designed surface is important for realization of the designed function. These results can be used to improve computational design strategies, but there is still much to be learned; for example, one designed complex, which does form in experiments, was classified by all metrics as a nonbinder. 相似文献
17.
Joanna Kamińska Olga M. Koper Edyta Siedlecka-Czykier Joanna Matowicka-Karna Jerzy Bychowski Halina Kemona 《Saudi Journal of Biological Sciences》2018,25(7):1263-1271
Introduction
Thrombotic and inflammatory mechanisms are involved in the pathophysiology of acute coronary syndrome (ACS). The aim of the study was the evaluation of inflammation (white blood cells count/WBC, C-reactive protein/CRP, interleukin-6/IL-6) and platelet (platelet count/PLT, mean platelet volume/MPV, large platelet/LPLT, beta-thromboglobulin/β-TG) biomarkers in the groups of ACS patients depending on the severity of signs and symptoms and compared to controls without coronary artery disease.Materials and methods
The study group included 93 patients categorized into 3 subgroups depending on the severity of signs and symptoms of ACS. PLT, MPV, LPLT, and WBC were determined on hematological analyzer, IL-6 and β-TG were measured using the ELISA method.Results
In the whole group of ACS patients WBC, CRP, IL-6, MPV, and β-TG were significantly higher as compared to controls. Analyzing the inflammation and platelet biomarkers depending on the severity of signs and symptoms in comparison to controls, statistically significant differences for above-mentioned parameters were also found. There were no significant differences between the advancement of coronary artery changes and inflammation as well as platelet parameters, except for CRP concentrations. The AUCs for all inflammation parameters tested were similar, however the highest AUCs showed WBC and CRP. Among platelet parameters the highest AUC revealed β-TG.Conclusion
Markers of inflammation and platelet activation may be associated to myocardial ischemia and myocardial injury. WBC, CRP and IL-6 as inflammation parameters and MPV and β-TG as platelet biomarkers may be useful indicators of the presence of coronary artery disease. 相似文献18.
Paul M. O Neill Paul A. Stocks Sunil Sabbani Natalie L. Roberts Richard K. Amewu Emma R. Shore Ghaith Aljayyoussi I?igo Angulo-Barturén María Belén Santiago Ferrer Bazaga María Santos Martínez Brice Campo Raman Sharma Susan A. Charman Eileen Ryan Gong Chen David M. Shackleford Stephen A. Ward 《Bioorganic & medicinal chemistry》2018,26(11):2996-3005
A series of aryl carboxamide and benzylamino dispiro 1,2,4,5-tetraoxane analogues have been designed and synthesized in a short synthetic sequence from readily available starting materials. From this series of endoperoxides, molecules with in vitro IC50s versus Plasmodium falciparum (3D7) as low as 0.84?nM were identified. Based on an assessment of blood stability and in vitro microsomal stability, N205 (10a) was selected for rodent pharmacokinetic and in vivo antimalarial efficacy studies in the mouse Plasmodium berghei and Plasmodium falciparum Pf3D70087/N9 severe combined immunodeficiency (SCID) mouse models. The results indicate that the 4-benzylamino derivatives have excellent profiles with a representative of this series, N205, an excellent starting point for further lead optimization studies. 相似文献
19.
《Chronobiology international》2013,30(3):331-347
With the introduction of sustained-release theophyllíne formulations for once-daily dosing or for unequally divided twice-daily dosing, comparison with conventional equally divided twice-daily dosing has been focused on nocturnal serum theophylline concentrations (STCs), plateau properties and peak-trough fluctuation. The merits of various steady-state characteristics such as nocturnal excess, plateau time, residual concentration, peak-trough fluctuation, swing and AUC fluctuation are illustrated by 15 data sets from 7 multiple-dose studies, each including either 10–12 healthy volunteers or 12–20 COPD-patients. In all of the studies, STCs were determined at least every 2 hr over a 24-hr period in steady-state. Included in the studies were 7 sustained-release theophylline formulations which were administered either once daily (in the morning or in the evening), or twice daily (either equally divided, or unequally divided with one-third of the dose being given in the morning and two-thirds in the evening). 相似文献
20.
Parkinson disease (PD) is the second most common neurodegenerative disorder after Alzheimer's disease, affecting 1–2% of the population over the age of 65. Both genetic and environmental factors trigger risks of and protection from PD. However, the molecular mechanism of PD is far from being clear. In this study, we downloaded the gene expression profile of PD from Gene Expression Omnibus and identified differentially expressed genes (DEGs) and dysfunctional pathways in PD patients compared with controls. To further understand how these pathways act together to account for the initiation of PD, we constructed a pathway crosstalk network by calculating the Jaccard index among pathways. A total of 873 DEGs and 16 dysfunctional pathways between PD patients and controls were identified. Through constructing a network of pathways, the relationships among PD pathways were visually presented by their interactions. Our results demonstrate the existence of crosstalk between different pathways in PD pathogenesis. These results not only may explain the causes of PD, but could also open the door to new therapeutic approaches for this disease. 相似文献