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91.
Intracellular ice formation (IIF) is almost invariably lethal. In most cases, it results from the too rapid cooling of cells to below −40 °C, but in some cases it is manifested, not during cooling, but during warming when cell water that vitrified during cooling first devitrifies and then recrystallizes during warming. Recently, Mazur et al. [P. Mazur, I.L. Pinn, F.W. Kleinhans, Intracellular ice formation in mouse oocytes subjected to interrupted rapid cooling, Cryobiology 55 (2007) 158–166] dealt with one such case in mouse oocytes. It involved rapidly cooling the oocytes to −25 °C, holding them 10 min, rapidly cooling them to −70 °C, and warming them slowly until thawed. No IIF occurred during cooling but intracellular freezing, as evidenced by blackening of the cells, became detectable at −56 °C during warming and was complete by −46 °C. The present study differs in that the oocytes were warmed rapidly from −70 °C to temperatures between −65 and −50 °C and held for 3–60 min. This permitted us to determine the rate of blackening as function of temperature. That in turn allowed us to calculate the activation energy (Ea) for the blackening process; namely, 27.5 kcal/mol. This translates to about a quadrupling of the blackening rate for every 5 °C rise in temperature. These data then allowed us to compute the degree of blackening as a function of temperature for oocytes warmed at rates ranging from 10 to 10,000 °C/min. A 10-fold increase in warming rate increased the temperature at which a given degree of blackening occurred by 8 °C. These findings have significant implications both for cryobiology and cryo-electron microscopy. 相似文献
92.
93.
Rondón LJ Groenestege WM Rayssiguier Y Mazur A 《American journal of physiology. Regulatory, integrative and comparative physiology》2008,294(6):R2001-R2007
The body maintains Mg(2+) homeostasis by renal and intestinal (re)absorption. However, the molecular mechanisms that mediate transepithelial Mg(2+) transport are largely unknown. Transient receptor potential melastatin 6 (TRPM6) was recently identified and shown to function in active epithelial Mg(2+) transport in intestine and kidney. To define the relationship between Mg(2+) status and TRPM6 expression, we used two models of hypomagnesemia: 1) C57BL/6J mice fed a mildly or severely Mg(2+)-deficient diet, and 2) mice selected for either low (MgL) or high (MgH) erythrocyte and plasma Mg(2+) status. In addition, the mice were subjected to a severely Mg(2+)-deficient diet. Our results show that C57BL/6J mice fed a severely Mg(2+)-deficient diet developed hypomagnesemia and hypomagnesuria and showed increased TRPM6 expression in kidney and intestine. When fed a Mg(2+)-adequate diet, MgL mice presented hypomagnesemia and hypermagnesuria, and lower kidney and intestinal TRPM6 expression, compared with MgH mice. A severely Mg(2+)-deficient diet led to hypomagnesemia and hypomagnesuria in both strains. Furthermore, this diet induced kidney TRPM6 expression in MgL mice, but not in MgH mice. In conclusion, as shown in C57BL/6J mice, dietary Mg(2+)-restriction results in increased Mg(2+) (re)absorption, which is correlated with increased TRPM6 expression. In MgL and MgH mice, the inherited Mg(2+) status is linked to different TRPM6 expression. The MgL and MgH mice respond differently to a low-Mg(2+) diet with regard to TRPM6 expression in the kidney, consistent with genetic factors contributing to the regulation of cellular Mg(2+) levels. Further studies of these mice strains could improve our understanding of the genetics of Mg(2+) homeostasis. 相似文献
94.
Termination of protein synthesis (hydrolysis of the last peptidyl-tRNA on the ribosome) takes place when the ribosomal A site is occupied simultaneously by one of the three stop codons and by a class-1 translation termination factor. The existing procedures to measure the functional activity of this factor both in vitro and in vivo have serious drawbacks, the main of which are artificial conditions for in vitro assays, far from those in the cell, and indirect evaluation of activity in in vivo systems. A simple reliable and sensitive system to measure the functional activity of class-1 translation termination factors could considerably expedite the study of the terminal steps of protein synthesis, at present remaining poorly known, especially in eukaryotes. We suggest a novel system to test the functional activity in vitro using native functionally active mRNA, rather than tri-, tetra-, or oligonucleotides as before. This mRNA is specially designed to contain one of the three terminating (stop) codons within the coding nucleotide sequence. Plasmids have been generated that carry the genes of suppressor tRNAs each of which is specific toward one of the three stop codons. They were shown to support normal synthesis of a reporter protein, luciferase, by reading through the stop codon within the coding mRNA sequence. We have demonstrated that human class-1 translation termination factor eRF1 is able to compete with suppressor tRNA for a stop codon and to completely prevent its suppressive effect at a sufficient concentration. Forms of eRF1 with point mutations in functionally essential regions have lower competitive ability, demonstrating the sensitivity of the method to the eRF1 structure. The enzymatic reaction catalyzed by the full-size reporter protein is accompanied by emission of light quanta. Therefore, competition between suppressor tRNA and eRF1 can be measured using a luminometer, and this allows precise kinetic measurements in a continuous automatic mode. 相似文献
95.
The efficiency of the wave energy loss from a nonuniform MHD waveguide due to the conversion of the trapped magnetosonic waveguide modes into runaway Alfvén waves is estimated theoretically. It is shown that, if the waveguide parameters experience a jumplike change along the waveguide axis, the interaction between the waveguide modes and Alfvén waves occurs precisely at this “jump.” This effect is incorporated into the boundary conditions. A set of coupled integral equations with a singular kernel is derived in order to determine the transmission and reflection coefficients for the waveguide modes. The poles in the kernels of the integral operators correspond to the surface waves. When the jump in the waveguide parameters is small, analytic expressions for the frequency dependence of the transformation coefficients are obtained by using a model profile of the Alfvén velocity along the magnetic field. For the jump characterized by the small parameter value ε=0.3, the wave-amplitude transformation coefficient can amount to 5–10%. Under the phase synchronization condition (when the phase velocities of the waveguide modes on both sides of the jump are the same), the wave-energy transformation coefficient is much higher: it increases from a fraction of one percent to tens of percent. The transformation of fast magnetosonic waves into Alfvén waves is resonant in character, which ensures the frequency and wavelength filteringof the emitted Alfvén perturbations. 相似文献
96.
M Zió?kowska-Kochan G Sobczak-Kamińska R Mazur 《Polski tygodnik lekarski (Warsaw, Poland : 1960)》1992,47(14-15):327-328
Many countries are interested in the cost of therapy of the acute cerebral flow insufficiency leading to brain infarction. Total costs include: drugs, laboratory tests, nursing care, the cost of patients' food, equipment, amortization, hospital administration etc. The total expenditure for a 10-15-day treatment of the acute neurological incident depended on the severity of the disease and was 8 million ++ z?oty in case of mildly ill patient, 9 million in case of moderately severe disease, and 21 million z?oty in case of severely ill patient. If the costs of such tests as CT and TCD-scanning will be added, total cost will increase significantly. 相似文献
97.
98.
Mathematical models of the transfer of large enough charged molecules (macroions) have been constructed on the basis of the
classical equations of electromigration diffusion (Helmholtz-Smoluchowski, Goldman, and Goldman-Hodgkin-Katz). It is shown
that ion transfer in placental barriers (mimicking lipid-protein membrane barriers) and in muscle barriers proceeds by different
mechanisms. In placental barriers, the electromigration diffusion takes place through lipid-protein channels formed by conformational
alteration of phospholipid and protein molecules, with diffusion coefficients D = (2.6–3.6) × 10−8 cm2/s. The transfer in muscle barriers is due to migration via charged interfibrillar channels with negative diffusion activation
energy (explained by changes in the structure of muscle fibers and expenditures of thermal energy for the displacement of
Cl− from channel walls), and D = (6.0–10.0) × 10−6 cm2/s. 相似文献
99.
100.