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81.
Transfer of allergic encephalomyelitis with spleen cells from donors sensitized with myelin basic protein in incomplete Freund's adjuvant 总被引:1,自引:0,他引:1
T Namikawa J R Richert B F Driscoll M W Kies E C Alvord 《Journal of immunology (Baltimore, Md. : 1950)》1982,128(2):932-934
Myelin basic protein (BP) emulsified in incomplete Freund's adjuvant (BP/IFA) is relatively nonencephalitogenic in Lewis rats. Furthermore, repeated injections of BP/IFA prevent subsequent induction of experimental allergic encephalomyelitis (EAE) by BP emulsified in complete Freund's adjuvant (BP/CFA). In spite of this, spleen cells from rats injected repeatedly with BP/IFA transfer EAE after they are cultured with BP almost as effectively as BP/CFA spleen cells. However, unlike the latter, BP/IFA spleen cells do not proliferate in response to BP in culture. Furthermore, BP/IFA spleen cells are unable to transfer EAE after culture with concanavalin A (Con A), in contrast to BP/CFA spleen cells. Both populations of spleen cells undergo a strong proliferative response to Con A in culture. For BP/IFA cells, at least, a proliferative response to BP in vitro is not a prerequisite for enhanced transfer of EAE in Lewis rats. 相似文献
82.
Distinctive effects of inhibitors of mitochondrial function on Rous sarcoma virus replication and malignant transformation 总被引:2,自引:0,他引:2
The effects of chloramphenicol and ethidium bromide on Rous sarcoma virus replication and transformation were studied. Chloramphenicol suppressed virus production with little effect on focus formation. Ethidium bromide inhibited focus formation up to 95% when cells were treated 24 hours prior to, or within the first 48 hours after infection, but failed to suppress virus replication significantly. These results suggest that virus replication and cell conversion require a function(s) having the antibiotic sensitivity resembling that of mitochondria. 相似文献
83.
The effects of a number of proteinase inhibitors on rat ovarian and rat hepatic adenylate cyclase preparations were examined. N alpha-tosylarginine methyl ester, 7-amino-1-chloro-3-L-tosylamidoheptan-2-one, 1-chloro-4-phenyl-3-L-tosylamidobutan-2-one, 1-chloro-4-methyl-3-L-tosylamidopentan-2-one and other low-molecular-weight proteinase inhibitors blocked hormonally stimulated adenylate cyclase from either source with hepatic preparations requiring higher concentrations. Addition of nucleotides (ATP, GTP, GDP, CTP or ITP) to inhibited ovarian preparations did not reverse inhibition, nor did dithiothreitol reverse phenylmethanesulphonyl fluoride-inhibited ovarian adenylate cyclase. The kinetics of the inhibition of rat ovarian adenylate cyclase were examined by following the production of cyclic AMP after the addition of inhibitors to membrane preparations preincubated under assay conditions with human choriogonadotropin, guanosine 5'-[beta gamma-imido]triphosphate of NaF. 7-Amino-1-chloro-3-L-tosylamidoheptan-2-one, 1-chloro-4-phenyl-3-L-tosylamidobutan-2-one and 1-chloro-4-methyl-3-L-tosylamidopentan-2-one had two effects on human-choriogonadotropin-stimulated adenylate cyclase. At low concentrations (less than or equal to 0.2 mM) there was an irreversible inhibition of hormonally-stimulated cyclase with maximum first-order inhibitory rate constants of 0.05--0.08 min-1. At higher concentrations the irreversible effect persisted, but, in addition, there was a marked decrease in the cyclase initial velocity to 25--50% of that of control values. N alpha-tosylarginine methyl ester had similar effects; at low concentrations (less than or equal to 2 mM) it inhibited irreversibly, and at higher concentrations it decreased the initial velocity (50% at 10 mM). At high concentrations (greater than 3 mM) N alpha-tosylarginine methyl ester also inhibited NaF- and guanosine 5'-[beta gamma-imidol]-triphosphate-stimulated cyclase but in a reversible manner. 7-Amino-1-chloro-3-L-tosylamidoheptan-2-one inhibited NaF-stimulated adenylate cyclase in two ways, as for human-choriogonadotropin-stimulated adenylate cyclase, but required 10--20-fold higher concentrations. The low-concentration irreversible effect can be explained by a continual inactive in equilibrium active conversion of adenylate cyclase during hormonal stimulation in which the inactive to active conversion is blocked by the inhibitors. The high-concentration effect is a direct one on the active catalytic moiety of the enzyme. 相似文献
84.
J R Richert B F Driscoll M W Kies E C Alvord 《Journal of immunology (Baltimore, Md. : 1950)》1979,122(2):494-496
Spleen cells from myelin basic protein (BP)-sensitized donor rats appear to be incapable of adoptively transferring experimental allergic encephalomyelitis (EAE) directly to normal recipients. It has been reported that in vitro incubation with concanavalin A (Con A) activates rat spleen cells so that they are capable of transferring EAE. We report here that incubation with specific antigen, BP, also permits transfer of disease with spleen cells. Data are presented in which activation of EAE spleen cells by Con A is compared with activation by BP. Cellular proliferation does not appear to be necessary for in vitro activation with specific antigen. 相似文献
85.
Orr S Alexandre E Clark BJ Gray N Helin H Ravid R Read M Richert L Roosipuu R Jonsson-Rylander AC Syring C Teesalu T Thasler WE Trafford J Van Der Valk J Weiss TS Womack C Ylikomi T 《Cell and tissue banking》2005,6(2):131-138
This report records the Fourth meeting of the European Network of Research Tissue Bank (Brussels, 18th March 2004) which was attended by Mel Read MEP. The existing membership of this informal group represents European Human Research Tissue Bankers, biomedical researchers seeking access to human tissue and allied groups including animal welfare representatives. This Fourth meeting provided a forum to update members on individual activity in this area. A particular focus of this meeting was to consider the status of this group and future affiliations to increase the profile and activity of this Network. This meeting addressed differences in legislative and ethical requirements governing the use of human tissue in biomedical research in the different countries represented. Future activity of the ENRTB, planned at this meeting, will target harmonisation of current differences which are currently barriers to increased access to human tissue for biomedical research. Through the harmonisation of procurement, processing and distribution of human tissue specimens the ENRTB will provide a mechanism to benefit human health through increased use of human tissue in pharmacotoxicological studies and the associated replacement of animal tests. 相似文献
86.
Elhaïmeur F Nicod L Courderot-Masuyer C Robin S Guyon C Bouhaddi M Regnard J Richert L Berthelot A 《Biological trace element research》2005,107(3):263-276
Hypertension is known to be associated with an oxidative stress resulting from an imbalance of antioxidant defense mechanisms
in various tissues. The purpose of this study was to investigate the relationship between the increase of arterial blood pressure,
measured during the gradual development of experimental hypertension in deoxycorticosterone (DOCA)-salt-treated rats, and
an early imbalance of liver antioxidant status. The levels of liver oxidant/antioxidant markers and iron were studied during
the induction of hypertension in 3-, 6-, and 8-wk DOCA-salt-treated Sprague-Dawley rats. Hepatic antioxidant defenses were
decreased as early as 3 wk of hypertensive treatment: the decrease of peroxidase-reductase-transferase and catalase activities
was associated with a significant increase of thiobarbituric acid reactive substances (TBARS) levels. Liver oxidative stress
increased until 6 wk, and remained stable at 8 wk of DOCA-salt treatment. Concurrently, liver iron levels were increased at
6 wk and returned to normal values after 8 wk of hypertensive treatment. Iron seems to be an inductor of liver oxidative stress
and responsible for the persistent oxidative stress, most likely through secondary free-radical release. Thus, our data (1)
confirm that hypertension in DOCA-salt-treated rats might be a free-radical-dependent disease where hepatic oxidant/antioxidant
imbalance is obviously involved from the beginning of blood pressure elevation and (2) suggest that the use of suitable iron
chelators might reverse liver oxidative stress associated with the increase of blood pressure. 相似文献
87.
Salah Edin El Meshri Emmanuel Boutant Assia Mouhand Audrey Thomas Valéry Larue Ludovic Richert Valérie Vivet-Boudou Yves Mély Carine Tisné Delphine Muriaux Hugues de Rocquigny 《Biochimica et Biophysica Acta (BBA)/General Subjects》2018,1862(6):1421-1431
Background
HIV-1 Gag polyprotein orchestrates the assembly of viral particles. Its C-terminus consists of the nucleocapsid (NC) domain that interacts with RNA, and the p6 domain containing the PTAP motif that binds the cellular ESCRT factor TSG101 and ALIX. Deletion of the NC domain of Gag (GagNC) results in defective Gag assembly, a decrease in virus production and, thus probably affects recruitment of the ESCRT machinery. To investigate the role of GagNC in this recruitment, we analysed its impact on TSG101 and ALIX localisations and interactions in cells expressing Gag.Methods
Cells expressing mCherry-Gag or derivatives, alone or together with eGFP-TSG101 or eGFP-ALIX, were analysed by confocal microscopy and FLIM-FRET. Chemical shift mapping between TSG101-UEV motif and Gag C-terminus was performed by NMR.Results
We show that deletion of NC or of its two zinc fingers decreases the amount of Gag-TSG101 interacting complexes in cells. These findings are supported by NMR data showing chemical shift perturbations in the NC domain in- and outside - of the zinc finger elements upon TSG101 binding. The NMR data further identify a large stretch of amino acids within the p6 domain directly interacting with TSG101.Conclusion
The NC zinc fingers and p6 domain of Gag participate in the formation of the Gag-TSG101 complex and in its cellular localisation.General significance
This study illustrates that the NC and p6 domains cooperate in the interaction with TSG101 during HIV-1 budding. In addition, details on the Gag-TSG101 complex were obtained by combining two high resolution biophysical techniques. 相似文献88.
Mutant KB cells with decreased EGF receptor expression: biochemical characterization 总被引:2,自引:0,他引:2
Mutants of the human KB carcinoma cell line resistant to a cytotoxic conjugate of epidermal growth factor and Pseudomonas exotoxin (EGF-PE) express a pleiotropic phenotype, which includes reduced levels of 125I-EGF binding, without altered affinity for EGF (Lyall et al., 1987). Here, the EGF-toxin (ET) resistant mutants were further characterized with respect to the amount and size of the EGF receptor and the level of EGF receptor RNA. These data indicate that decreased binding of 125I-EGF in the mutants is due to reduced amounts of EGF receptor, which is associated with decreased mRNA levels. Changes in other proteins in the ET mutants were also examined. Five of the six ET mutants had a decrease in a 78,000 Mr- membrane glycoprotein. In addition, an increase in a protein with a Mr- of 40,000 and a pl = 8.0 was found in all the mutants, and an increase in a series of proteins with a Mr- of 36,000 and a pl of 6.3-6.5 was found in some of the mutants. These results confirm the pleiotropic nature of the EGF-PE resistant mutants and show that reduced EGF binding is due to altered expression of the EGF receptor gene in the mutants. 相似文献
89.
D W Shen C Cardarelli J Hwang M Cornwell N Richert S Ishii I Pastan M M Gottesman 《The Journal of biological chemistry》1986,261(17):7762-7770
We have established four cell lines derived from the human KB carcinoma cell line which express high-level multiple drug resistance. One of these lines was selected for resistance to colchicine, one was selected for resistance to colchicine in the presence of the tumor promoter, mezerein, one for resistance to vinblastine, and one for resistance to adriamycin. All of these cell lines are cross-resistant to the other selective agents. The development of multidrug resistance in these cultured human carcinoma cells is associated with a limited number of specific protein alterations revealed by high resolution two-dimensional gel electrophoresis and Western blot analysis. These protein alterations in multidrug-resistant lines include the decreased prevalence of members of a family of proteins of molecular mass 70,000 to 80,000 daltons, pI 4.8-5.0, the increased synthesis of a protein of molecular mass 21,000 daltons, pI 5.0, in the colchicine-resistant cell lines only, and the increased expression of a 170,000-dalton protein in membrane preparations from all of the resistant cells. The loss of the 70,000- to 80,000-dalton proteins in the multidrug-resistant lines, which can also be demonstrated by immunoprecipitation of these proteins with specific antisera, is associated with a loss of translatable mRNA for these proteins. These studies suggest that only a limited number of protein changes occur in multidrug-resistant cell lines. 相似文献
90.
The phosphorylation of vinculin by a highly purified kinase was stimulated by anionic phospholipids and inhibited to varying degrees, by chlorpromazine, imipramine, dibucaine and tetracaine. The drug effects are ascribed to a competitive inhibition of the activation process by their ability to interact with phospholipid. 相似文献