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81.
High‐affinity aptamers for important signal transduction proteins, i.e. Cdc42‐GTP, p21‐activated kinase1 (PAK1) and MRCK (myotonic dystrophy kinase‐related Cdc42‐binding kinase) α were successfully selected in the low micro‐ to nanomolar range using non‐systematic evolution of ligands by exponential enrichment (SELEX) with at least three orders of magnitude enhancement from their respective bulk affinity of naïve DNA library. In the non‐SELEX procedure, CE was used as a highly efficient affinity method to select aptamers for the desired molecular target through a process that involved repetitive steps of partitioning, known as non‐equilibrium CE of equilibrium mixtures with no PCR amplification between successive steps. Various non‐SELEX conditions including the type, concentration and pH of the run buffer were optimized. Other considerations such as salt composition of selection buffer, protein concentration and sample injection size were also studied for high stringency during selection. After identifying the best enriched aptamer pool, randomly selected clones from the aptamer pool were sequenced to obtain the individual DNA sequences. The dissociation constants (Kd) of these sequences were in the low micromolar to nanomolar range, indicating high affinity to the respective proteins. The best binders were also subjected to sequence alignment to generate a phylogenetic tree. No significant consensus region based on approximately 50 sequences for each protein was observed, suggesting the high efficiency of non‐SELEX for the selection of numerous unique sequences with high selectivity. 相似文献
82.
King‐Fu Lin Jing‐Yau Shyu 《Journal of polymer science. Part A, Polymer chemistry》2001,39(18):3085-3092
The early cure behavior of 4,4‐dicyanato 1,1‐diphenolethane resin with and without incorporating Cr(acac)3, Co(acac)3, and Cu(acac)2, respectively, as catalysts was investigated by gel permeation chromatography. The curing intermediates were separated by the column elution method and characterized by Fourier transform infrared, 1H, and 3C NMR spectroscopies. The results indicated that the formed dimer in the early cure stage is a straight chain containing a primary amino group. The formed triazine ring in the trimer has a strong catalytic effect on the remaining cyanate groups so that the reactivity of the trimers was significantly increased. The reactivities of the curing intermediates decreased with molecular size until 7‐mer was reached. The initial monomer consumption is described by second‐order‐rate kinetics. In the presence of metal acetylacetonates, the curing reactions may be accelerated, but they did not change the reaction path and preceding sequence of reactivities. © 2001 John Wiley & Sons, Inc. J Polym Sci Part A: Polym Chem 39: 3085–3092, 2001 相似文献
83.
This report describes a method to simultaneously determine 11 low-molecular-weight (LMW) organic acids and 16 chlorinated acid herbicides within a single run by a portable CE system with contactless conductivity detection (CCD) in a poly(vinyl alcohol) (PVA)-coated capillary. Under the optimized condition, the LODs of CE-CCD ranged from 0.056 to 0.270 ppm, which were better than for indirect UV (IUV) detection of the 11 LMW organic acids or UV detection of the 16 chlorinated acid herbicides. Combined with an on-line field-amplified sample stacking (FASS) procedure, sensitivity enhancement of 632- to 1078-fold was achieved, with satisfactory reproducibility (RSDs of migration times less than 2.2%, and RSDs of peak areas less than 5.1%). The FASS-CE-CCD method was successfully applied to determine the two groups of acidic pollutants in two kinds of environmental water samples. The portable CE-CCD system shows advantages such as simplicity, cost effectiveness, and miniaturization. Therefore, the method presented in this report has great potential for onsite analysis of various pollutants at the trace level. 相似文献
84.
Yau‐Hung Chen Chien‐Hui Chiou Wei‐Li Chen Yu‐Ru Jhou Ya‐Ting Lee Chien‐Chung Cheng 《中国化学会会志》2010,57(6):1257-1261
Zebrafish have received considerable attention as an organism‐based model in the development of pharmacological agents.1,2 Many small molecules applied to zebrafish show important behaviours and may constitute new kinds of markers for clinical purposes.3 Analysis of these molecules can facilitate the development of useful tools for monitoring environmental changes.4 Many chemicals that are toxic to the environment are known to influence the sensory systems of humans5 and fish.6 One important sensory system in all fish is the lateral line organ,7 which is readily accessible for the assessment of environmental changes.8 Neuromasts, which are located on the surface of the fish body, are one of the major components of the lateral lines of the zebrafish.9 Copper‐enriched water is known to affect the olfactory system in fish. Therefore, small molecules that induce specific patterns in the neuromasts of zebrafish should provide an important animal model with which to explore the effects of environmental changes on the sensory system.10,11 Recently, chemical sensors based on the rhodamine skeleton12 have been designed to specifically detect metal ions, such as Cu(II)13 and Fe(III)/Hg(II),14 in zebrafish. However, there has been no report of these rhodamine derivatives used in the specific recognition of the sensory system of zebrafish. Commonly, the sensory system is studied with antibody staining assays of scarified fish. Here, we report that a new rhodamine derivative can be used as a fluorescent chemical probe to visualize the neuromasts and intestinal villi of living zebrafish. Based on the specific recognition of this area in zebrafish, we narrowed the possible enzymes targeted by this rhodamine probe to alkaline phosphatase and confirmed this with a binding assay. It is a well‐recognized challenge to develop a fluorescent chemical probe that specifically recognizes a particular enzyme. Furthermore, the transfer of phosphate groups to certain enzymes can activate their catalytic reactivity, triggering a cascade reaction in a signal transduction pathway. The alkaline‐phosphatase‐specific recognition by this rhodamine derivative may be applicable to clinical purposes. 相似文献
85.
Various phase behavior of blends of poly(vinyl ether)s with homologous acrylic polymers (polymethacrylates or polyacrylates) were examined using differential scanning calorimetry, optical microscopy (OM), and Fourier‐transformed infrared spectroscopy. Effects of varying the pendant groups of either of constituent polymers on the phase behavior of the blends were analyzed. A series of interestingly different phase behavior in the blends has been revealed in that as the pendant group in the acrylic polymer series gets longer, polymethacrylate/poly(vinyl methyl ether) (PVME) blends exhibit immiscibility, upper critical solution temperature (UCST), and miscibility, respectively. This study found that the true phase behavior of poly(propyl methacrylate)/PVME [and poly(isopropyl methacrylate)/PVME)] blend systems, though immiscible at ambient, actually displayed a rare UCST upon heating to higher temperatures. Similarly, as the methyl pendant group in PVE is lengthened to ethyl (i.e., PVME replaced by PVEE), phase behavior of its blends with series of polymethacrylates or polyacrylates changes correspondingly. Analyses and quantitative comparisons on four series of blends of PVE/acrylic polymer were performed to thoroughly understand the effects of pendant groups in either polyethers (PVE's) or acrylic polymers on the phase behavior of the blends of these two constituents. © 2007 Wiley Periodicals, Inc. J Polym Sci Part B: Polym Phys 45: 1521–1534, 2007 相似文献
86.
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89.
The thermal ionization rates of electrons and holes at cobalt centers in silicon have been measured using the dark-capacitance and dark-current transient techniques. Using only n+–p diodes, the cobalt charge states, the thermal emission rates including their field dependences are determined. The thermal emission rate data fitted to et = A exp (−ΔE/kT) gives: (i) ΔE = ET − EV = (377 ± 1) meV and A = (4.43 ± 0.33) × 1013 s−1 for the donor level, (ii) ΔE = EC − ET = (569 ± 3) meV and A = (2.59 ± 0.38) × 1012 s−1 for the acceptor level. 相似文献
90.
Motivated by mirror symmetry, we study certain integral representations of solutions to the Gel´fand-Kapranov-Zelevinsky (GKZ) hypergeometric system. Some of these solutions arise as period integrals for Calabi-Yau manifolds in mirror symmetry. We prove that for a suitable compactification of the parameter space, there exist certain special boundary points, which we called maximal degeneracy points, at which all solutions but one become singular.