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Summary NAD(P)H fluorescence emission spectra are recorded from single living cells, by a recently developed multichannel microspectrofluorometric technique, in correlation with the intracellular microelectrophoretic addition of substrate (i. e., glucose-6-P). These spectra may be used as a reference basis in establishing the critical parameters to be followed when the same studies are extended to a variety of cells, submitted to various drug effects or physical treatments. The sum-spectra corresponding to channel by channel (wavelength by wavelength) summation of spectra obtained from various cells within a series, before and after addition of substrate, and their difference spectrum may be normalized and evaluated comparatively. The NAD(P)H emission maximum prior to addition of substrate seems to present a mixture of dehydrogenase-bound and free coenzyme. There is a suggestion that immediately after substrate, i. e., at 5 sec, an increase in free NADH is first observed. While the overall changes in fluorescence intensity associated with substrate are quite large (50–150% increase), the counts (i. e., an expression of photons) associated with shifts in the emission maximum (free vs. bound NAD(P)H changes) are at times barely above noise. Rapid microspectrofluorometry provides in principle the most direct approach for the identification of coenzyme bound to various dehydrogenases in single living cells, but further improvements in spectral resolution and signal-to-noise ratio are required, for a better definition of the spectral shifts which may be observed.
Zusammenfassung Mit Hilfe einer kürzlich entwickelten Mehrkanal-Mikrospektrofluorometer-Methode wurden von einzelnen lebenden Zellen nach intrazellulär mikroelektrophoretischer Substratzugabe (z. B. Glucose-6-P) NAD(P)H Fluoreszenz-Emissionsspektren aufgenommen. Diese Spektren können als Vergleichsbasis bei der Festsetzung der entscheidenden Parameter verwendet werden, wenn die gleichen Untersuchungen auf eine Reihe von Zellen ausgedehnt werden sollen, die verschiedenen Medikamenteffekten oder physikalischer Behandlung ausgesetzt werden. Die Summenspektren, die der kanalmäßigen (wellenlängenmäßigen) Summierung der Spektren von verschiedenen Zellen innerhalb einer Serie, vor und nach Substratzugabe entsprechen, sowie ihre Differenzspektren können normalisiert und vergleichsweise ausgewertet werden. Das NAD(P)H-Emissionsmaximum vor der Substratzugabe scheint ein Gemisch von freiem und dehydrogenasegebundenem Coenzym darzustellen. Unmittelbar nach Substratzugabe (d. h. nach 5 sec) ist ein Anstieg an freiem NADH das erste Mal zu beobachten. Während die mit dem Substrat einhergehenden Gesamtveränderungen der Fluoreszenzintensität recht groß sind (50–150% Anstieg), sind die Impulse (als Effekt der Photonen), die mit einer Verschiebung im Emissionsmaximum verbunden sind (Veränderungen von freiem und gebundenem NAD(P)H) zu manchen Zeiten kaum höher als das Rauschen. Die rasche Mikrospektrofluorometrie stellt im Prinzip die direkteste Methode zur Identifizierung von Coenzymen dar, die an verschiedenen Dehydrogenasen in einzelnen lebenden Zellen gebunden sind. Weitere Verbesserungen der Spektralauflösung und der Empfindlichkeit (signal-to-noise ratio) sind notwendig, um die Spektralverschiebungen, die beobachtet werden, besser auswerten zu können.相似文献
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Jocelyn Jalbert Roland Gilbert Pierre Tétreault Brigitte Morin Denise Lessard-Déziel 《Cellulose (London, England)》2007,14(4):295-309
In this study, headspace gas chromatography/mass spectrometry has been used to assess the volatile by-products generated by
the ageing of oil-impregnated paper insulation of power transformers. Sealed-glass ampoules were used to age under oxidative
conditions 0.5-g specimens of insulating paper in 9 mL of inhibited mineral oil in a temperature range of 60–120 °C and moisture
of 0.5, 1 and 2% (w/w). A linear relationship between one of the oil-soluble degradation by-products, i.e. methanol, and the
number of ruptured 1,4-β-glycosidic bonds of cellulose, regardless of the type of paper (ordinary Kraft or thermally-upgraded
(TU) Kraft paper), was established for the first time in this field. Ageing at 130 °C of model compounds of the Kraft paper
constituents (α-cellulose, hemicellulose and lignin) and two cellulosic breakdown by-products (D-(+)-glucose and 1,6-anhydro-β-d-glucopyranose) confirmed that the α-cellulose degradation was mostly responsible for the presence of this molecule in the
system. Furthermore, additional 130 °C-tests with six different papers and pressboard samples under a tight control of initial
moisture indicated that at least one molecule of methanol is formed for each rupture of 1,4-β-glucosidic bond of the molecular
chains. Stability tests showed that the ageing indicator is stable under the oxygen and temperature conditions of open-breathing
transformers. The presence of methanol was detected in 94% of oil samples collected from over than 900 in-service pieces of
equipment, confirming the potential for this application. Lastly, the tests have shown that oil-oxidation by-products and
TU-nitrogenous agents modify the methanol partitioning coefficients in the paper/oil/air system, which makes their study essential
over a range of field conditions encountered by power transformers. Results are presented and discussed in comparison with
2-furfuraldehyde, which is the current reference in the domain. 相似文献
119.
Fadila Balegroune Pierre Braunstein Laurent Douce Yves Dusausoy Daniel Grandjean Michael Knorr Martin Strampfer 《Journal of Cluster Science》1992,3(3):275-296
The reactions of K[Fe{Si(OMe)3}(CO)3(PY)][PY=Ph2PCH2C(O)Ph, Ph2PCH2C(O)[(-C5H4)FeCp] (Cp=5-C5H5), Ph2P(CH2)2CN] with CdCl2·2.5H2O, ZnX
2 (X=Cl, I) or InCl3 afforded Fe-Cd-Fe or Fe-M(-X)2
M-Fe (M=Cd, Zn, In;X=Cl, I) and Fe-InCl2 complexes. Some of them contain an unusual and labile -2-SiO alkoxysilyl bridge which may be associated with a bridging mode for the ketophosphine ligand (first such example structurally established), thus providing original results in bimetallic chemistry on the intramolecular coordination of oxygendonor functions ofchemically different hemilabile ligands firmly attached to a neighboring metal center. The structures of the trinuclear complex (3), of the chlorobenzene solvate of the tetranuclear complex (4a·C6H5Cl) and of [mer-(OC)3{(EtO)3Si} (4e) have been determined by X-ray diffraction. Crystals of 3 are orthorhombic, space groupPbcn, witha=19.010(4),b=11.766(5),c=26.998(7)Å, andZ=4. Crystals of4a·C6H5Cl are monoclinic, space groupC2/c witha=22.455(3),b=17.680(2),c=16.627(4)Å, =90.80(4)°, andZ=4. Crystals of4e are monoclinic, space groupC2/c witha=25.392(5),b=18.554(6),c=16.28(1)Å, =120.73(3)°, andZ=4. The structures were solved using direct methods and Fourier difference techniques and refined by blocked full-matrix least squares toR=0.035 (R
w
=0.049) for 2719 observed reflections, toR=0.042 (R
w
=0.056) for 3082 observed reflections, and toR=0.057 (R
w
=0.075) for 1850 observed reflections for3, 4a·C6H5Cl and4e, respectively. The Fe-Zn complexes (9a), (9b) and (9c) were prepared and characterized by spectroscopic methods.Part 21 in the Series: Complexes with Functional Phosphines. Part 20: P. Braunstein, S. Coco Cea, A. DeCian, and J. Fischer (1992).Inorg. Chem.
31, 4203. 相似文献
120.
Jean Luc Guermond 《Zeitschrift für Angewandte Mathematik und Physik (ZAMP)》1987,38(5):717-729
Asymptotic theories like the lifting-line, the slender body or the slender ship lead to lineintegrals with singular kernels. Sometimes these integrals are improper, that is to say that they are defined only by their Finite Part. To find asymptotic expansions of these integrals, the Matched Asymptotic Expansion Method is widely used along with other more specific methods depending on the kernel type. The first method is laborious and not systematic, and the other methods are sometimes too much specific to treat general cases. Moreover, all of them are not well adapted to deal with Finite Part integrals.Here, a new method is proposed to avoid the previous difficulties. This method is systematic for homogeneous kernels and gives approximations up to any order, as long as the derivative of the weight function exists at this given order. Moreover the occurrence of logarithmic terms in the expansion is explained and easily predictable. An elliptic integral and the classical lifting-line theory are treated to illustrate the ease of this method.
Nomenclature D domain of integration - f(x) weight function - FP Finite Part - h(x) weight function - I ,I o bounded intervals - j, J integers - K(x, ) singular kernel - L, L integers - M integer defining the approximation order - P k (x) Legendre polynomial - R set of real numbers - R(ß) equals 1 if is an integer and 0 if not - R f, J ,R K, L remainders of Taylor developments - S () equals either 1 or the sign function:sgn() - t, u, v, x variable of integration - , real numbers - homogeneity order of the kernel - F () Euler's integral (gamma function) - small parameter - [.] integer part of 相似文献
Résumé Les théories asymptotiques telles que la ligne portante, le corps élancé ou le navire de grand allongement conduisent à des intégrales curvilignes à noyaux singuliers. Parfois, ces intégrales sont impropres c'est à dire qu'elles sont définies en Parties Finies. Différentes méthodes ont été mises au point pour trouver les développements asymptotiques de ces intégrales. Généralement elles dépendent fortement de la nature du noyau, et c'est finalement la méthode des développements raccordés qui est utilisées quand le noyau est trop compliqué. Cependant, cette méthode est laborieuse et comme les précèdentes non adaptée aux intégrales défines par leur Partie Finie.Une nouvelle méthode est proposée pour surmonter ces difficultés. Cette méthode est systématique pour les noyaux homogènes et donne les approximations à tout ordre pourvu que les dérivées de la fonction poids existent jusqu'à cet ordre. De plus la présence de termes logarithmiques dans le développement est expliquée et aisément prédictible.Une intégrale elliptique, ainsi que la fameuse théorie de la ligne portante sont traités pour illustrer les possibilités de la méthode.
Nomenclature D domain of integration - f(x) weight function - FP Finite Part - h(x) weight function - I ,I o bounded intervals - j, J integers - K(x, ) singular kernel - L, L integers - M integer defining the approximation order - P k (x) Legendre polynomial - R set of real numbers - R(ß) equals 1 if is an integer and 0 if not - R f, J ,R K, L remainders of Taylor developments - S () equals either 1 or the sign function:sgn() - t, u, v, x variable of integration - , real numbers - homogeneity order of the kernel - F () Euler's integral (gamma function) - small parameter - [.] integer part of 相似文献