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931.
932.
Development of immobilized Sn4+ affinity chromatography material for highly selective enrichment of phosphopeptides 下载免费PDF全文
In this work, we first immobilized tin(IV) ion on polydopamine‐coated magnetic graphene (magG@PDA) to synthesize Sn4+‐immobilized magG@PDA (magG@PDA‐Sn4+) and successfully applied the material to highly selective enrichment of phosphopeptides. The material gathered the advantages of large surface area of graphene, superparamagnetism of Fe3O4, good hydrophilicity and biocompatibility of polydopamine, and strong interaction between Sn4+ and phosphopeptides. The enrichment performance of magG@PDA‐Sn4+ toward phosphopeptides from digested β‐casein at different concentrations, with and without added digested BSA was investigated and compared with magG@PDA‐Ti4+. The results showed high selectivity and sensitivity of the Sn4+‐IMAC material toward phosphopeptides, as good as the Ti4+‐IMAC material. Finally, magG@PDA‐Sn4+ was applied to the analysis of endogenous phosphopeptides from a real sample, human saliva, with both MALDI‐TOF MS and nano‐LC‐ESI‐MS/MS. The results indicated that the as‐synthesized Sn4+‐IMAC material not only has good enrichment performance, but also could serve as a supplement to the Ti4+‐IMAC material and expand the phosphopeptide coverage enriched by the single Ti4+‐IMAC material, demonstrating the broad application prospects of magG@PDA‐Sn4+ in phosphoproteome research. 相似文献
933.
Duong T Nguyen Baojun Wu Hongan Long Nan Zhang Caitlyn Patterson Stephen Simpson Krystalynne Morris W Kelley Thomas Michael Lynch Weilong Hao 《Molecular biology and evolution》2020,37(11):3118
Mutation and recombination are the primary sources of genetic variation. To better understand the evolution of genetic variation, it is crucial to comprehensively investigate the processes involving mutation accumulation and recombination. In this study, we performed mutation accumulation experiments on four heterozygous diploid yeast species in the Saccharomycodaceae family to determine spontaneous mutation rates, mutation spectra, and losses of heterozygosity (LOH). We observed substantial variation in mutation rates and mutation spectra. We also observed high LOH rates (1.65–11.07×10−6 events per heterozygous site per cell division). Biases in spontaneous mutation and LOH together with selection ultimately shape the variable genome-wide nucleotide landscape in yeast species. 相似文献
934.
PC12 cell line is well documented and widely applied as many kinds of models in neurobiological and neurochemical studies. Yet a thorough proteomic analysis has not been performed so far. Here we report the construction of a large-scale 2-D protein database for PC12 cells. The proteins extracted from PC12 cells were separated by 2-DE and identified by MALDI-TOF/TOF MS. A total of 1080 protein spots, excised from three different 2-D gels, were identified with high confidence. These proteins represent 474 different gene products, mainly binding proteins and enzymes. Three hundred and seven identified protein spots were located in the low-molecular weight region below 20 kDa. This database today represents one of the largest 2-D databases for higher eukaryotic cell proteomes and for low-molecular weight proteins. In addition, fragment ion spectra obtained by TOF/TOF confirmed that calcylin in PC12 cells was N-acetylated. The database of PC12 proteome is expected to be a powerful tool for neuroscientists. 相似文献
935.
Linlin Zhao Matthew G. Pence Plamen P. Christov Zdzislaw Wawrzak Jeong-Yun Choi Carmelo J. Rizzo Martin Egli F. Peter Guengerich 《The Journal of biological chemistry》2012,287(42):35516-35526
N2,3-Ethenoguanine (N2,3-ϵG) is one of the exocyclic DNA adducts produced by endogenous processes (e.g. lipid peroxidation) and exposure to bioactivated vinyl monomers such as vinyl chloride, which is a known human carcinogen. Existing studies exploring the miscoding potential of this lesion are quite indirect because of the lability of the glycosidic bond. We utilized a 2′-fluoro isostere approach to stabilize this lesion and synthesized oligonucleotides containing 2′-fluoro-N2,3-ϵ-2′-deoxyarabinoguanosine to investigate the miscoding potential of N2,3-ϵG by Y-family human DNA polymerases (pols). In primer extension assays, pol η and pol κ replicated through N2,3-ϵG, whereas pol ι and REV1 yielded only 1-base incorporation. Steady-state kinetics revealed that dCTP incorporation is preferred opposite N2,3-ϵG with relative efficiencies in the order of pol κ > REV1 > pol η ≈ pol ι, and dTTP misincorporation is the major miscoding event by all four Y-family human DNA pols. Pol ι had the highest dTTP misincorporation frequency (0.71) followed by pol η (0.63). REV1 misincorporated dTTP and dGTP with much lower frequencies. Crystal structures of pol ι with N2,3-ϵG paired to dCTP and dTTP revealed Hoogsteen-like base pairing mechanisms. Two hydrogen bonds were observed in the N2,3-ϵG:dCTP base pair, whereas only one appears to be present in the case of the N2,3-ϵG:dTTP pair. Base pairing mechanisms derived from the crystal structures explain the slightly favored dCTP insertion for pol ι in steady-state kinetic analysis. Taken together, these results provide a basis for the mutagenic potential of N2,3-ϵG. 相似文献
936.
Kesheng Lin Zhengqi Wang Li Wang Jiawen Zhou Lijuan Han Jingjing Wen Tingxia Dong Ran Duan Ning Li 《Phyton》2023,92(2):423-437
Prohibited pesticide residues have become one of the main factors affecting the quality and safety of Lycii Fructus, However, rarely studies focus on the rapid determination of these residues. Here, a total of 30 kinds of prohibited pesticide residues were determined by ultra-performance liquid chromatography tandem triple quadrupole mass spectrometry (UPLC-MS/MS) in five different process ways. Pretreatment methods, chromatographic separation and detection conditions in mass spectrometry were all optimized accordingly. Among the five different pretreatment methods, the first and third solid phase extraction failed to provide high recoveries of sulfosulfuron compounds (both lower than 60%). Recovery of chlorphenamidine by the Quick Easy Cheap Effective Rugged and Safe multiresidue method (QuEChERS) was lower than 60%, which did not meet the requirements of trace determination. The concentrations of 30 prohibited pesticides residues treated by straightforward and solid phase extraction showed good linearity in their corresponding ranges, with correlation coefficients over 0.99. The average recoveries of straightforward ranged from 78.13% to 110.9%, while RSD ranged from 1.3% to 16.9%, albeit poor purification was observed. The recovery yield from solid phase extraction was between 67.75% and 103.08% with RSD value from 0.8% to 14.0%, which met the requirements of trace determination, this method has good precision and stability. These results could be employed to other Traditional Chinese Medicines (TCMs) in detecting prohibited pesticide residues. 相似文献
937.
938.
疏叶当归根的化学成分 总被引:7,自引:0,他引:7
从四川茂汶产的疏叶当归根的乙醇提取物中分离得9个化合物、鉴定了7个,分别为当归内酯、β-谷甾醇、β-胡萝卜甙、地十作碳酸、蔗糖、10-庚基-10-辛基-十二烷及疏叶香豆素,其中疏叶香豆素为一新化合物,对钙拮抗有一定活性,另2个长链脂肪族化合物未鉴定。这9个化合物是从该植物中立冬以。疏叶当归根的乙醚提取物,经气-质联用分析得10个挥发性成分,主含当归内酯(65%),还含少量的δ(1-甲基乙基乙烯基) 相似文献
939.
A quantitative proteomics analysis of MCF7 breast cancer stem and progenitor cell populations 下载免费PDF全文
Song Nie Sean P. McDermott Yadwinder Deol Zhijing Tan Max S. Wicha David M. Lubman 《Proteomics》2015,15(22):3772-3783
Accumulating evidence has demonstrated that breast cancers are initiated and develop from a small population of stem‐like cells termed cancer stem cells (CSCs). These cells are hypothesized to mediate tumor metastasis and contribute to therapeutic resistance. However, the molecular regulatory networks responsible for maintaining CSCs in an undifferentiated state have yet to be elucidated. In this study, we used CSC markers to isolate pure breast CSCs fractions (ALDH+ and CD44+CD24‐ cell populations) and the mature luminal cells (CD49f‐EpCAM+) from the MCF7 cell line. Proteomic analysis was performed on these samples and a total of 3304 proteins were identified. A label‐free quantitative method was applied to analyze differentially expressed proteins. Using the criteria of greater than twofold changes and p value <0.05, 305, 322 and 98 proteins were identified as significantly different in three pairwise comparisons of ALDH+ versus CD44+CD24‐, ALDH+ versus CD49f‐EpCAM+ and CD44+CD24‐ versus CD49f‐EpCAM+, respectively. Pathway analysis of differentially expressed proteins by Ingenuity Pathway Analysis (IPA) revealed potential molecular regulatory networks that may regulate CSCs. Selected differential proteins were validated by Western blot assay and immunohistochemical staining. The use of proteomics analysis may increase our understanding of the underlying molecular mechanisms of breast CSCs. This may be of importance in the future development of anti‐CSC therapeutics. 相似文献
940.