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41.
Reshu Saxena Sudipti Gupta Kavita Singh Kalyan Mitra Anil Kumar Tripathi Raj Kamal Tripathi 《PloS one》2015,10(4)
Nef is an accessory viral protein that promotes HIV-1 replication, facilitating alterations in cellular pathways via multiple protein-protein interactions. The advent of proteomics has expanded the focus on better identification of novel molecular pathways regulating disease progression. In this study, nef was sequenced from randomly selected patients, however, sequence variability identified did not elicited any specific mutation that could have segregated HIV-1 patients in different stages of disease progression. To explore the difference in Nef functionality based on sequence variability we used proteomics approach. Proteomic profiling was done to compare the effect of Nef variants in host cell protein expression. 2DGE in control and Nef transfected SupT1 cells demonstrated several differentially expressed proteins. Fourteen protein spots were detected with more than 1.5 fold difference. Significant down regulation was seen in six unique protein spots in the Nef treated cells. Proteins were identified as Cyclophilin A, EIF5A-1 isoform B, Rho GDI 1 isoform a, VDAC1, OTUB1 and α-enolase isoform 1 (ENO1) through LC-MS/MS. The differential expression of the 6 proteins was analyzed by Real time PCR, Western blotting and Immunofluorescence studies with two Nef variants (RP14 and RP01) in SupT1 cells. There was contrasting difference between the effect of these Nef variants upon the expression of these six proteins. Downregulation of α-enolase (ENO1), VDAC1 and OTUB1 was more significant by Nef RP01 whereas Cyclophilin A and RhoGDI were found to be more downregulated by Nef RP14. This difference in Nef variants upon host protein expression was also studied through a site directed mutant of Nef RP01 (55AAAAAAA61) and the effect was found to be reversed. Deciphering the role of these proteins mediated by Nef variants will open a new avenue of research in understanding Nef mediated pathogenesis. Overall study determines modulation of cellular protein expression in T cells by HIV-1 Nef variants. 相似文献
42.
N transfer in three-species grass-clover mixtures with chicory,ribwort plantain or caraway 总被引:1,自引:0,他引:1
Nawa Raj Dhamala Jim Rasmussen Georg Carlsson Karen Søegaard Jørgen Eriksen 《Plant and Soil》2017,413(1-2):217-230
Background and aims
There is substantial evidence that legume-derived Nitrogen (N) is transferred to neighboring non-legumes in grassland mixtures. However, there is sparse information about how deep rooted non-legume forage herbs (forbs) influence N transfer in multi-species grasslands.Methodology
Red clover (Trifolium pretense L.) was grown together with perennial ryegrass (Lolium perenne L.) and one of three forb species: chicory (Cichorium intybus L.), ribwort plantain (Plantago lanceolata L.) or caraway (Carum carvi L.) in a field experiment. During the first year after the establishment, red clover leaves were labeled with 15N-urea to determine the N transfer from red clover to companion ryegrass and forbs.Results
On an annual basis, up to 15 % of red clover N was transferred to the companion ryegrass and forbs, but predominantly to the grass. The forb species did not differ in their ability to take up clover N, but biomass production and soil N acquisition was higher in chicory and plantain than in caraway.Conclusions
Grass relied to a great extent on clover N, whereas forbs relied on soil N. Soil 15N-enrichment indicated that N transfer occurred in the upper soil layers and that a dependence on clover-derived N did not necessarily give grass a growth advantage.43.
Jagesh K. Tiwari Poonam D. Sarkar SK. Pandey Jai Gopal S. Raj Kumar 《Plant Cell, Tissue and Organ Culture》2010,103(2):175-187
Interspecific potato somatic hybrids between Solanum tuberosum L. (di)haploid C-13 and 1 endosperm balance number non-tuberous wild species S. etuberosum Lindl. were produced by protoplasts electrofusion. The objective was to transfer virus resistance from this wild species
into the cultivated potatoes. Post-fusion products were cultured in VKM medium followed by regeneration of calli in MS13 K medium at 20°C under a 16-h photoperiod, and regenerants were multiplied on MS medium. Twenty-one somatic hybrids were
confirmed by RAPD, SSR and cytoplasm (chloroplast/mitochondria) type analysis possessing species-specific diagnostic bands
of corresponding parents. Tetraploid nature of these somatic hybrids was determined through flow cytometry analysis. Somatic
hybrids showed intermediate phenotypes (plant, leaves and floral morphology) to their parents in glass-house grown plants.
All the somatic hybrids were male-fertile. ELISA assay of somatic hybrids after artificial inoculation of Potato virus Y (PVY)
infection reveals high PVY resistance. 相似文献
44.
45.
Mechanism of action of interleukin-13 antagonist (IL-13E13K) in cells expressing various types of IL-4R 总被引:6,自引:0,他引:6
As interleukin (IL)-13 and IL-4 play a major role in various diseases including asthma, allergy, and malignancies, it is desirable to generate a molecule that blocks the effects of both cytokines. We previously generated a human IL-13 mutant (IL-13E13K), which is a powerful antagonist of IL-13, blocking the biological activities of IL-13. We now show that IL-13E13K also competitively inhibits signaling and biological activities of IL-4 through type II and partially through type III IL-4 receptor (R) system. IL-13E13K completely blocked the IL-4-induced phosphorylation of STAT6 and IL-4-dependent protein synthesis in cells expressing type II and partially type III IL-4R but not type I IL- 4R. Consistent with the inhibition of biological activities, IL-13E13K inhibited IL-4 binding to type II IL-4R-expressing cells but not to type I IL-4R-expressing cells. The inhibition efficiency of IL-4 binding by IL-13E13K was relatively lower compared to wtIL-13 even though IL-13E13K bound to IL-13Ralpha1 positive cells with a similar affinity to wtIL-13. These results indicate that Glu13 in IL-13 associates with IL-4Ralpha, and mutation to lysine decreases its binding ability to IL-4Ralpha chain. IL-13E13K binds to IL- 13Ralpha1, which is shared by both IL-13R and IL-4R systems. Consequently, IL-13E13K inhibits IL-4 binding to these cells and prevents heterodimer formation between IL-13Ralpha1 and IL-4Ralpha chains. This interference by IL-13E13K blocks the biological activities of not only IL-13 but also partially of IL-4. Thus, IL-13E13K may be a useful agent for the treatment of diseases such as asthma, allergic rhinitis, and cancer, which are dependent on signaling through both IL-4 and IL-13 receptors. 相似文献
46.
47.
48.
Differential modulation of voltage-dependent K+ currents in colonic smooth muscle by oxidants 总被引:1,自引:0,他引:1
The effect of oxidants on voltage-dependent K+ currents was examined in mouse colonic smooth muscle cells. Exposure to either chloramine-T (Ch-T), an agent known to oxidize both cysteine and methionine residues, or the colon-specific oxidant monochloramine (NH2Cl) completely suppressed the transient outward K+ current (Ito) while simultaneously enhancing the sustained delayed rectifier K+ current (Idr). In contrast, the cysteine-specific oxidants hydrogen peroxide (H2O2) and 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB) exhibited partial and slow suppression of Ito by inducing a shift in channel availability of -18 mV without affecting Idr. After enhancement by NH2Cl or Ch-T, Idr was sensitive to 10 mM tetraethylammonium but not to other K+ channel blockers, suggesting that it represented activation of the resting Idr and not a separate K+ conductance. Extracellular dithiothreitol (DTT) partially reversed the effect of H2O2 and DTNB on Ito but not the actions of NH2Cl and Ch-T on either Idr or Ito. Dialysis of myocytes with GSH (5 mM) or DTT (5 mM) prevented suppression of Ito by H2O2 and DTNB but did not alter the effects of NH2Cl or Ch-T on either Idr or Ito. Ch-T and NH2Cl completely blocked Ito generated by murine Kv4.1, 4.2, and 4.3 in Xenopus oocytes, an effect not reversible by intracellular DTT. In contrast, intracellular DTT reversed the effect of H2O2 and DTNB on the cloned channels. These results suggest that Ito is suppressed via modification of both methionine and cysteine residues, whereas enhancement of Idr likely results from methionine oxidation alone. colon; colitis; redox; ion channel 相似文献
49.
50.
Arginine is one of the commonly used additives to enhance refolding yield of proteins, to suppress aggregation of proteins, and to increase solubility of proteins, and yet the molecular interactions that contribute to the role of arginine are unclear. Here, we present experiments, using bovine serum albumin (BSA), lysozyme (LYZ), and β-lactoglobulin (BLG) as model proteins, to show that arginine can enhance heat-induced aggregation of concentrated protein solutions, contrary to the conventional belief that arginine is a universal suppressor of aggregation. Results show that the enhancement in aggregation is caused only for BSA and BLG, but not for LYZ, indicating that arginine's preferential interactions with certain residues over others could determine the effect of the additive on aggregation. We use this previously unrecognized behavior of arginine, in combination with density functional theory calculations, to identify the molecular-level interactions of arginine with various residues that determine arginine's role as an enhancer or suppressor of aggregation of proteins. The experimental and computational results suggest that the guanidinium group of arginine promotes aggregation through the hydrogen-bond-based bridging interactions with the acidic residues of a protein, whereas the binding of the guanidinium group to aromatic residues (aggregation-prone) contributes to the stability and solubilization of the proteins. The approach, we describe here, can be used to select suitable additives to stabilize a protein solution at high concentrations based on an analysis of the amino acid content of the protein. 相似文献