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111.
Dulce E. Oliveira Ana Lúcia C. Santos Neto Anita D. Panek 《Analytical biochemistry》1981,113(1):188-192
A quantitative in situ assay of yeast α-glucosidase involving permeabilization of the cells by freezing and thawing is described. The assay was applied to different strains in different physiological states and was shown to give results comparable to those obtained with total cell homogenates. The primary advantage of the in situ assay was the possibility of analyzing a large number of samples from the same culture during a growth curve using a very reduced cell mass. 相似文献
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The isolation of Saccharomyces cerevisiae plasma membrane was carried out after hypotonic lysis of yeast protoplasts treated with concanavalin A by two independent methods: a, at low speed centrifugation and b, at high speed centrifugation in a density gradient. Several techniques (electron microscopic, enzymic, tagging, etc.) were used to ascertain the degree of purification of the plasma membranes obtained. The low speed centrifugation technique as compared with the other method gave a higher yield of plasma membranes with a similar degree of purification. Analysis of the yeast plasma membrane of normally growing cells by sodium dodecyl sulphate polyacrylamide gel electrophoresis showed at least 25 polypeptide bands. Twelve glycoprotein bands were also found, and their apparent molecular weights were determined. Treatment of the protoplasts with cycloheximide resulted in a significant decrease in the carbohydrate and protein content of the plasma membrane. The electrophoretic pattern of the plasma membrane of cycloheximide-treated cells showed a redistribution of the relative amounts of each protein band and a drastic reduction in the number of Schiff-positive bands. The isoelectric point of the most abundant proteins was low (pI 4) or lower than expected from previous data. A large part of the mannosyl transferase activity found in the cell (80%) was associated with the internal membranes, the remaining activity (20%) was located in the plasma membrane preparation. Part of the mannosyl transferase activity of the cells is located at the plasma membrane surface. Invertase (an external mannoprotein) is found in both the plasma and internal membranes, and as the specific activity dropped significantly following cycloheximide treatment of the cells, it is suggested that these membranes systems are the structures for the glycosylation of a precursor invertase and its subsequent release into the periplasmic space. Other transferase found in the plasma membrane preparation transfers glucose residues from UDPglucose to a poly(alpha(1 leads to 4) polymer identified as glycogen. 相似文献
115.
J D Ashbrook A A Spector E C Santos J E Fletcher 《The Journal of biological chemistry》1975,250(6):2333-2338
The binding of six physiologically important long chain fatty acids to defatted human plasma albumin was measured at 37 degrees in a calcium-free Krebs-Ringer phosphate buffer, pH 7.4. The data were analyzed in terms of multiple stepwise equilibria. With the saturated acids, the magnitude of the equilibrium (association) constants, Ki, increased as the chain length increased: laurate smaller than myristate smaller than palmitate smaller than stearate. Oleate was bound more tightly than stearate; by contrast, linoleate was bound less tightly than stearate. The equilibrium constants, K1 through K12, ranged from 2.4 times 10-6 - 3.5 times 10-3 m-1 for laurate to 2.6 times 10-8 - 3.5 times 10-5 m-1 for oleate. Successive values of Ki decrease for each of the acids, indicating that major cooperative binding effects do not occur over the physiological range of fatty acid concentrations. In no case could the Ki be segregated into distinct classes, suggesting that any grouping of albumin binding sites is somewhat arbitrary. The results were inconclusive concerning whether premicellar association of unbound fatty acid occurs. Although corrections for premicellar association produced very little change in the Ki values for myristate, they raised the Ki for palmitate and stearate by 300 to 700 per cent. A sigmoidal relationship was obtained when the logarithm of Ki was plotted against chain length for the saturated fatty acids containing 6 to 18 carbon atoms, indicating that the binding energy is not simply a statistical process dependent only on the fatty acid chain length. This selectivity that albumin contributes to the binding process may be due to varying degrees of configurational adaptability of its binding sites as the fatty acid increases in length. 相似文献
116.
P G Santos F Grande 《Proceedings of the Society for Experimental Biology and Medicine. Society for Experimental Biology and Medicine (New York, N.Y.)》1975,149(3):652-655
The lipolytic effect of glucagon was measured in vitro with adipose tissue of "young" (4-8 wk) and "old" (over 1 yr) geese. The response of the young geese tissue was about twice that observed with tissue of old geese, for glucagon concentrations of 0.05, 0.5, and 5.0 mug/ml. Our estimates indicate that the number of adipose cells per g of adipose tissue of young geese was three times that of the old geese tissue. This suggests that the greater lipolytic response to glucagon, observed in young geese adipose tissue, may possibly be due to its greater cellularity, rather than to a greater lipolytic response of the individual adipocyte. The lipolytic effect of glucagon in vivo, for each of the doses between 1.0 and 20.0 mug/kg, was significantly greater in the old than in the young geese. The slope of the linear equation relating log10 of glucagon dose and elevation of plasma FFA 5 min after injection, was significantly greater for the old than for the young geese. In the goose, therefore, the influence of age on the adipokinetic effect of glucagon appears to be mediated by factors operating in the whole animal, more than by changes in the adipose cell itself. A slower removal rate of circulating FFA by the old geese, could be one of these factors. 相似文献
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Raquel Santos Jorge Troy R. Hawkins Edgar G. Hertwich 《The International Journal of Life Cycle Assessment》2012,17(1):9-15
Purpose
The purpose of this study is to provide life cycle inventory data and results for components of electrical grids to the larger community of life cycle assessment practitioners. This article is the first in a series of two, each focusing on different components of power grids. In part 1, the objects under scope are power lines and cables. Systems for overhead, underground, and subsea transmission are modeled here, including HVDC systems used in long-distance transmission. 相似文献120.
Leonardo Mata Helena Gaspar Fátima Justino Rui Santos 《Journal of applied phycology》2011,23(5):827-832
The genus Asparagopsis is a prolific source of halogenated metabolites. Due to its commercial applications, it has been intensively cultivated in
southern Portugal. In the present study, we assess if the internal levels of the major halogenated metabolites (bromoform
and dibromoacetic acid) in Asparagopsis taxiformis can be increased with hydrogen peroxide (H2O2) addition. Previous studies with red algae showed that the production/release of bromoform can be enhanced by exogenously
supplying H2O2. However, no study has assessed if H2O2 supply enhances the content of secondary metabolites within the biomass. This detail is important as the objective of the
proposed research is to enhance the content of these valuable metabolites in the produced biomass. Both the activity of the
haloperoxidase enzyme and the metabolite content were assessed on short-term and long-term incubation periods to H2O2. To determine the susceptibility of A. taxiformis photosynthetic performance to the imposed oxidative stress, the in vivo fluorescence of photosystem II was monitored. A. taxiformis was shown to be physiologically vulnerable to H2O2, given the observed decrease of the maximum quantum yield of photosynthesis (F
v/F
m). Contrary to what was expected, the presence of H2O2 inhibited the activity of the iodoperoxidase enzyme. Nevertheless, the extracted halogenated metabolites were higher over
the first hours of exposure to H2O2, decreasing after 48 h. These results are probably related to the prosthetic group of the halogenated enzyme in A. taxiformis and the long-term oxidative stress damage of H2O2 exposure. Considering the objective of the proposed research, addition of H2O2 to the cultures, prior (3 h) to biomass harvesting, increases the metabolite content. 相似文献