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21.
K-Cl cotransport activity in frog erythrocytes was estimated as a Cl- -dependent component of K+ efflux from cells incubated in Cl- - or NO3- -containing medium at 20 degrees C. Decreasing the osmolality of the medium resulted in an increase in K+ efflux from the cells in a Cl- medium but not in an NO3- medium. Treatment of red cells with 5 mM NaF caused a significant decrease (approximately 50%) in K+ loss from the cells in iso- and hypotonic Cl- media but only a small decrease in K+ loss in isotonic NO3- medium. Addition of 1 mM vanadate to an isotonic Cl- medium also led to a significant reduction in K+ efflux. Similar inhibitory effects of NaF and vanadate on K+ efflux in a Cl- medium, but not in an NO3- medium were observed when the incubation temperature was decreased from 20 to 5 degrees C. Thus, under various experimental conditions, NaF and vanadate inhibited about 50% of Cl- -dependent K+ efflux from frog red cells probably due to inhibition of protein phosphatases. Cl- -dependent K+ (86Rb) influx into frog erythrocytes was nearly completely blocked (approximately 94%) by 5 mM NaF. In a NO3- medium, K+ influx was mainly mediated by the Na+,K+ pump and was unchanged in the presence of 5 mM NaF, 0.03 mM Al3+ or their combination. These data indicate that G proteins or cAMP are not involved in the regulation of Na+,K+ pump activity which is activated by catecholamines and phosphodiesterase blockers in these cells. 相似文献
22.
23.
Gennadii Petrovich Gusev Natalia Ivanovna Agalakova 《Journal of comparative physiology. B, Biochemical, systemic, and environmental physiology》2010,180(3):385-391
Recently (Agalakova and Gusev in J Comp Physiol 179:443–450, 2009), we demonstrated that the activity of K–Cl cotransport (KCC) in frog red blood cells is inhibited under stimulation of protein
kinase C (PKC) with phorbol ester PMA (12-myristate-13-acetate). Present work was performed to uncover possible implication
of protein kinases and protein phosphatases (PPs) in the regulation of baseline and volume-dependent KCC activity in these
cells. K+ influx was estimated as 86Rb uptake by the cells in isotonic or hypotonic media in the presence of ouabain, K+ efflux was determined as the difference between K+ loss by the cells incubated in parallel in isotonic or hypotonic K+-free Cl−- and NO3
−-media. Swelling of the cells in hypotonic medium was accompanied by approximately 50% activation of Cl-dependent K+ influx and efflux. Protein tyrosine kinase (PTK) inhibitor genistein (0.1 mM) stably and considerably (up to 89%) suppressed
both baseline and volume-dependent KCC activity in each direction. Other PTK blockers (tyrphostin 23 and quercetin) had no
influence on KCC activity in frog erythrocytes. PKC inhibitor chelerythrine (20 μM) and both PP inhibitors, fluoride (5 mM)
and okadaic acid (1 μM), reduced KCC activity by 25–70%. Neither basal nor swelling-activated KCC in frog erythrocytes was
affected by PKC inhibitor staurosporine (1 μM). Based on the previous and present results, we can suggest that the main role
in the maintenance of basal and volume-dependent KCC activity in frog erythrocytes belongs to PTKs and PPs, whereas PKC is
a negative regulator of this ion system. 相似文献
24.
Transport of 204Tl was studied in human erythrocytes incubated in isotonic salt solutions at pH 7.4 and 37 degrees C. 204Tl was rapidly accumulated in cells up to the constant level within a 10 minutes incubation (t0.5 = 3.5 min). The rate of uptake and the distribution ratio decreased in the presence of 0.1 mM ouabain and 0.5-1.0 mM furosemide (t0.5 = 5 min). A broad variability of the coefficient 204Tl distribution was observed in the intact erythrocytes due to a ouabain-sensitive component which was seen to decrease with the increase in Tl+ concentration in the medium (0.005-0.2 mM), and also to depend on the medium ion composition. On the contrary, a passive distribution of 204Tl in the presence of ouabain and furosemide was relatively constant within 1.1-1.5. The steady state distribution of 204Tl was declined after a substitution of Cl- by sucrose in the medium due to depolarization of erythrocyte membrane. On the other side, 204Tl uptake by the cells was raised during hyperpolarization of the membrane in the presence of valinomycin. 相似文献
25.
T. I. Ivanova A. O. Sherstobitov G. P. Gusev 《Journal of Evolutionary Biochemistry and Physiology》2007,43(6):557-563
To activate Na+/H+ exchange, intracellular pH (pHi) of erythrocytes of the river lamprey Lampetra fluviatilis were changed from 6 and 8 using nigericin. The Na+/H+ exchanger activity was estimated from the values of amiloride-sensitive components of Na+ (22Na) inflow or of H+ outflow from erythrocytes. Kinetic parameters of the carrier functioning were determined by using Hill equation. Dependence of Na+ and H+ transport on pHi value is described by hyperbolic function with the Hill coefficient value (n) close to 1. Maximal rate of ion transport was within the limits of 9–10 mmol/l cells/min, and the H+ concentration producing the exchanger 50% activation amounted to 0.6–1.0 μM. Stimulation of H+ outcome from acidified erythrocytes (pHi 5.9) with increase of H+ concentration in the incubation medium is described by Hill equation with n value of 1.6. Concentration Na+ for the semimaximal stimulation of H+ outcome amounted to 10 mM. The obtained results indicate the presence in lamprey erythrocytes of only binding site for H+ from the cytoplasm side and the presence of positive cooperativity in Na+-binding from the extracellular side of the Na+/H+ exchanger. Na+ efflux from cells in the Na+-free medium did not change at a 10-fold increase of H+ concentration in the incubation medium. The presented data indicate differences of kinetic properties of the lamprey erythrocyte Na+/H+ exchanger and of this carrier isoforms in mammalian cells. In intact erythrocytes the dependence of the amiloride-sensitive Na+ inflow on its concentration in the medium is described by Hill equitation with n 1.6. The Na+ concentration producing the 50% transport activation amounted to 39 mM and was essentially higher as compared with that in acidified erythrocytes. These data confirm conception of the presence of two amiloride-sensitive pathways of Na+ transport in lamprey erythrocytes. 相似文献
26.
27.
Riapis LA Filatov NN Salova NIa Sizykh EV Gerasimov AN Svetoch EA Stepanshin IuG Bannov VA Eruslanov BV Borzenkov VN Khramov MV Gusev VV 《Zhurnal mikrobiologii, epidemiologii, i immunobiologii》2005,(1):7-11
The characterization of E.coli strains O157:H7, isolated from humans and animals on some territories of the Central Federal District, is presented. Among the isolates from human outbreaks, related and, probably, related cultures prevailed, while among the isolates obtained from different animals mainly unrelated cultures have been detected. A conclusion has been made concerning the existence of several independent zoonotic reservoirs of E. coli O157:H7 infection on this territory. The advantages and drawbacks of the use of pulse electrophoresis in the characterization of E. coli O157:H7 are discussed. Grounds are given for the necessity of the patients examination with hemorrhagic enetrocolitis for the presence of E. coli O157:H7, as well as for the expediency of having a special item for the registration of this E. coli infection in relevant statistical forms. 相似文献
28.
Divergence patterns of the banding sequences from the chromosomal arms A, C, D, E, and F were compared in 63 species of the genus Chironomus. Evaluation of the number of breakpoints between the pairs of inverted banding sequences and the analysis of the lengths of the conserved segments in the chromosomal arms in the chironomid species examined showed that different arms evolved relatively independently and at different rates. No direct correlation between the arm length and the breakpoints number was observed. The length of the conservative segment was not fixed, but was arm-specific. Robustness and fidelity of the estimates of phylogenetic relationships between the species examined increased with the arm number, i.e., with the genome proportion included in the analysis. 相似文献
29.
Ivanova TI Agalakova NI Gusev GP 《Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology》2006,145(1):60-67
Four structurally different protein phosphatases (PPs) inhibitors - fluoride, calyculin A, okadaic acid and cantharidin--were tested for their ability to modulate unidirectional Na(+) influx in rat red blood cells. Erythrocytes were incubated at 37 degrees C in isotonic and hypertonic media containing 1 mM ouabain and (22)Na in the absence or presence of PP inhibitors. Exposure of the cells to 20 mM fluoride or 50 nM calyculin A for 1 h under isosmotic conditions caused a significant stimulation of Na(+) influx, whereas addition of 200 microM cantharidin or 100 nM okadaic acid had no effect. After 2 h of treatment, however, all these PPs blockers significantly enhanced Na(+) transport in rat erythrocytes. Selective inhibitors of PP-1 and PP-2A types, calyculin A, cantharidin and okadaic acid, produced similar ( approximately 1.2-1.4-fold) stimulatory effects on Na(+) influx in the cells. Activation of Na(+) influx was unchanged with increasing calyculin A concentration from 50 to 200 nM. No additive stimulation of Na(+) influx was observed when the cells were treated with combination of 20 mM fluoride and 50 nM calyculin A. Na(+) influx induced by PPs blockers was inhibited by 1 mM amiloride and 200 muM bumetanide approximately in the equal extent, indicating the involvement of Na(+)/H(+) exchange and Na-K-2Cl cotransport in sodium transport through rat erythrocytes membrane. Activation of Na(+) transport in the cells induced by calyculin A and fluoride was associated with increase of intracellular Na(+) content. Shrinkage of the rat erythrocytes resulted in 2-fold activation of Na(+) influx. All tested PPs inhibitors additionally activated the Na(+) influx by 70-100% above basal shrinkage-induced level. Amiloride and bumetanide have diminished both the shrinkage-induced and PPs-inhibitors-induced Na(+) influxes. Thus, our observations clearly indicate that activities of Na(+)/H(+) exchanger and Na-K-2Cl cotransporter in rat erythrocytes are regulated by protein phosphatases and stimulated when protein dephosphorylation is inhibited. 相似文献
30.