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991.
Zhao Y Kan ZY Zeng ZX Hao YH Chen H Tan Z 《Journal of the American Chemical Society》2004,126(41):13255-13264
Nucleic acid molecules may fold into secondary structures, and the formation of such structures is involved in many biological processes and technical applications. The folding and unfolding rate constants define the kinetics of conformation interconversion and the stability of these structures and is important in realizing their functions. We developed a method to determine these kinetic parameters using an optical biosensor based on surface plasmon resonance. The folding and unfolding of a nucleic acid is coupled with a hybridization reaction by immobilization of the target nucleic acid on a sensor chip surface and injection of a complementary probe nucleic acid over the sensor chip surface. By monitoring the time course of duplex formation, both the folding and unfolding rate constants for the target nucleic acid and the association and dissociation rate constants for the target-probe duplex can all be derived from the same measurement. We applied this method to determine the folding and unfolding rate constants of the G-quadruplex of human telomere sequence (TTAGGG)(4) and its association and dissociation rate constants with the complementary strand (CCCTAA)(4). The results show that both the folding and unfolding occur on the time scale of minutes at physiological concentration of K(+). We speculate that this property might be important for telomere elongation. A complete set of the kinetic parameters for both of the structures allows us to study the competition between the formation of the quadruplex and the duplex. Calculations indicate that the formation of both the quadruplex and the duplex is strand concentration-dependent, and the quadruplex can be efficiently formed at low strand concentration. This property may provide the basis for the formation of the quadruplex in vivo in the presence of a complementary strand. 相似文献
992.
993.
The covalent immobilization of DNA onto self-assembled monolayer (SAM) modified gold electrodes (SAM/Au) was studied by X-ray photoelectron spectrometry and electrochemical method so as to optimize its covalent immobilization on SAMs. Three types of SAMs with hydroxyl, amino, and carboxyl terminal groups, respectively, were examined. Results obtained by both X-ray photoelectron spectrometry and cyclic voltammetry show that the largest covalent immobilization amount of dsDNA could be gained on hydroxyl-terminated SAM/Au. The ratio of amount of dsDNA immobilized on hydroxyl-terminated SAMs to that on carboxyl-terminated SAMs and to that on amino-terminated SAMs is (3-3.5): (1-1.5): 1. The dsDNA immobilized covalently on hydroxyl-terminated SAMs accounts for 82.8-87.6% of its total surface amount (including small amount of dsDNA adsorbed). So the hydroxyl-terminated SAM is a good substrate for the covalent immobilization of dsDNA on gold surfaces. 相似文献
994.
A strain ofRhodococcus equi SHB-121 forming 3-cyanopyridine hydratase was screened from nitrile-polluted soil. The optimum conditions for the formation
of 3-cyanopyridine hydratase by the strain SHB-121 have been studied. Under the optimum conditions, the specific activity
of the enzyme reached 5.32 U/mg of dry cell, 95 times higher than that cultured in screening medium. In addition, the activity
of coexistent amidase was very low.
3-Cyanopyridine hydratase was purified from methylacrylamide-induced cells ofRh. equi SHB-121 by procedures including ultrasonic oscillation, ammonium sulfate precipitation, and column chromatographies on DEAE-cellulose
DE52, hydroxyapatite, and Sephadex G-25. The overall purification was 31-fold. The molecular weight of the enzyme was about
30 kDA by SDS-PAGE. The pI value was 4.1. The transition temperature and pH were 7.0°C and 6.0, respectively, resulting from
the differential spectra.
The optimum pH and temperature for the enzyme reaction were 8.0 and 30°C. The enzyme activity was strongly inhibited by Ag+, Hg2+, Cu2+, and NH4
+, whereas it was enhanced by Fe3+ slightly. The enzyme catalyzed the hydration of 3-cyanopyridine to nicotinamide, and itsKm value was 0.1 mol/L. Uncompetitive inhibitor sodium cyanide has a K, value of 5 mmol/L. 相似文献
995.
用ICP研究鹿科动物骨质疏松症的微量元素特征 总被引:1,自引:0,他引:1
鹿科动物的骨质疏松症是近几年出现的一种疾病,它影响鹿的生长发育和繁殖,病重者死亡。我们采集梅花鹿和白唇鹿的毛,血样,用低温灰化方法和温法消解处理样品,用ICP分析了毛,血清中微量元素的含量。病梅花鹿毛样中Mg,Al,Ca,Ti,Fe,Cu,Zn的含量比正常组高,Ti,Cu,Zn的偏差率大于15%,而Si,Mn,P,P,Ag,Ba,La,Ce,Pb比正常组低,V,Ag,Ba,La,Ce,Pb存在... 相似文献
996.
997.
本文基于二氧硫对固定于增塑的聚氯乙烯膜上的芳香稠环化合物蒽的荧光熄灭,研制了二氧化硫选择性光化学敏感膜,经过组成优化 的敏感膜测定二氧化硫的浓度范围为6.59×10^-3-5.23×10^-1mol/L,校正曲线与理论推导的模拟曲线相吻合,且荧光信号具有较高的重现性,信号响应时间为5-30s. 相似文献
998.
Zhao S Prasad C Robertson HJ Liu YM 《Fresenius' Journal of Analytical Chemistry》2001,369(3-4):220-224
A capillary electrophoresis (CE) method with laser induced fluorescence (LIF) detection is described for quantification of enterostatin (Val-Pro-Asp-Pro-Arg), a pentapeptide involved in appetite regulation and insulin secretion. Enterostatin and two other pentapeptides belonging to the enterostatin family (i.e. Ala-Pro-Gly-Pro-Arg and Val-Pro-Gly-Pro-Arg) were well separated from each other. The peptides were fluorescently tagged with naphthalene-2,3- dicarboxaldehyde (NDA) and separated by micellar electrokinetic chromatography (MEKC) in the presence of methanol as an organic modifier. Coupled with LIF detection, the method had a detection limit of 4.8 x 10(-6) M for enterostatin. The relative standard deviation was to be 4.0% from five determinations of enterostatin at 37.2 microM in a human cerebrospinal fluid (CSF) sample. Twenty-three human CSF samples were analyzed. The level of enterostatin ranged from 24 microM to 51 microM with a mean (+/- SEM) value of 41.7 +/- 2.0 microM. 相似文献
999.
The combination of matrix isolation infrared spectroscopic and density functional calculation results provides strong evidence that the transition metal monoxide cation, ScO+, coordinates five noble gas atoms in forming the [ScO(Ng)5]+ (Ng = Ar, Kr, or Xe) complexes in noble gas matrixes. 相似文献
1000.
双波长分光光度法同时测定贵金属钯和锇 总被引:4,自引:0,他引:4
本文提出了在对羟基苯基荧光酮和溴化十六烷基三甲烷基甲胺存在下,双波长分光光度法同时测定微量和锇的新方法,在pH=7.6~8.0的磷酸盐介质中,钯和锇的对羟基苯基荧光酮-CTMAB配合物吸收光谱相互重叠,选择测定钯配合物的波长对为580.5nm与530.5nm,测定锇配合物的波长对为553.5nm与604.5nm,钯含量在0~7.0μg/10mL范围内,锇含量在0~9.0μg/mL范围内,与ΔA值呈 相似文献