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1.
Initiation of the extrinsic pathway of coagulation. Association of factor VIIa with a cell line expressing tissue factor 总被引:3,自引:0,他引:3
We have examined initial assembly of the extrinsic pathway of blood coagulation on cell surfaces with radiolabeled human factor VIIa and a human fetal lung cell line possessing abundant functional tissue factor activity. Binding of factor VIIa to these cells was observed and was time- and temperature-dependent. Binding of factor VIIa was quantitatively equivalent at 37 and 6 degrees C, although the kinetics of binding differed. The radiolabeled ligand bound by the cell was indistinguishable by sodium dodecyl sulfate-polyacrylamide gel analysis from the factor VIIa offered. Factor VIIa binding was influenced by calcium ions. The binding appears to involve at least two classes of calcium-dependent binding sites. Optimal binding occurred at 2 mM calcium for both classes of sites, and there was inhibition of binding to the high affinity sites at higher calcium. Association of factor VIIa was specific, saturable, had a Kd of 123 +/- 37 pm, and factor VIIa interacted with about 100,000 binding sites per cell. Once established, specific binding was rapidly reversible. Direct cellular binding of human factor X also was observed and was calcium, time- and temperature-dependent. Factor X binding was specific and saturable with half-maximal binding at 87.6 +/- 27.4 nM to 6.03 +/- 1.03 X 10(6) sites per cell. Specific high affinity binding of factor VIIa correlated with generation of factor Xa. A direct linear relationship was observed at low factor VIIa binding; however, at higher bound factor VIIa, the relationship was nonstoichiometric, i.e. less factor Xa was formed per mole of factor VIIa. Expression of specific binding sites for factors VIIa and X provides further substantiation for the molecular assembly hypothesized to initiate the extrinsic coagulation protease cascade on cells. 相似文献
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E F Plow C Hougie T S Edgington 《Journal of immunology (Baltimore, Md. : 1950)》1971,107(5):1496-1500
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Y. BATARD A. ZIMMERLIN M. LE RET F. DURST D. WERCK-REICHHART 《Plant, cell & environment》1995,18(5):523-533
O-Dealkylation of two series of fluorescent 7-alkoxy-coumarins and 7-alkoxyphenoxazones by plant cytochrome P450s was investigated in Helianthus tuberosus tuber tissues treated with prototype P450 inducers, environmental pollutants or agrochemicals. Methoxy-, ethoxy-, propoxy-and butoxycoumarins and methoxy- and ethoxyresorufins were metabolized by fplant microsomes. Dealkylation of pentoxy- and benzyloxyresorufins was not detected. All dealkylating activities were enhanced by aging plant tissues in the presence of xenobiotics, in some cases up to 20-fold relative to the activities detected in control tissues. Increases in total P450 in the same tissues never exceeded 3-fold. The isozymes induced by prototype P450 inducers clearly differed from those in mammalian liver. That multiple P450s with overlapping substrate specificities were involved in the metabolism of both alkoxycoumarins and alkoxyresorufins was demonstrated by (1) the differential induction of the activities in response to exposure to xenobiotics, (2) the differential inhibition of the activities by clotrimazole, paclobutrazole and tetcyclacis in aminopyrine and benzo(a)pyrene-treated tissues, and(3) the selective inhibition observed with antibodies raised against purified ethoxycoumarin deethylase fractions. Our results suggest that the measurement of the dealkylation of such fluorescent substrates in plants might be useful to monitor environmental pollution. 相似文献
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Biotypes of Agrobacterium tumefaciens in Hungary 总被引:3,自引:1,他引:2
S. SÜLE 《Journal of applied microbiology》1978,44(2):207-213
Isolates of Agrobacterium tumefaciens from Hungary were separated into three biotypes on the basis of their physiological characters. Biotypes 1 and 2 corresponded with those of Keane et al . (1970). The most common isolates were of biotype 2. Isolates from grapevines formed a separate biotype which might be distinguished from biotype 1 by D-(–)tartrate and malonate utilization. Many isolates with biotype-intermediate characters were found. Isolates utilizing D-(–)tartrate, erythritol and malonate were included into biotype 2, although many of them were 3–ketolactose positive. Biotypes were not separated geographically and biotype 1 and 2 apparently occurred together. 相似文献
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