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Abstract— Pretreatment with sulfhydryl-reactive agents, such as N-ethylmaleimide and p-chloromercuriphenylsul-fonic acid, invariably resulted in marked inhibition of the binding of dl -(E)-2-amino-4-[3H]propyl-5-phosphono-3-pentenoic acid ([3H]CGP 39653), a competitive antagonist at an N-methyl-d -aspartate (NMDA)-sensitive subclass of central excitatory amino acid receptors, in brain synaptic membranes extensively washed and treated with Triton X-100, but did not significantly affect the binding of L-[3H]-glutamic acid ([3H]Glu), an endogenous agonist. The pre-treatment was effective in reducing the binding of [3H]-CGP 39653 at equilibrium, without altering the initial association rate, and decreased the affinity for the ligand. Pretreatment with sulfhydryl-reactive agents also enhanced the potencies of NMDA agonists to displace [3H]-CGP 39653 binding and attenuated those of NMDA antagonists, but had little effect on the potencies of the agonists and antagonists to displace [3H]Glu binding. The binding of both [3H]CGP 39653 and [3H]Glu was similarly sensitive to pretreatment with four different proteases in Tritontreated membranes, whereas pretreatment with phospho-lipase A2 or C markedly inhibited [3H]CGP 39653 binding without altering [3H]Glu binding. Moreover, both phospho-lipases not only induced enhancement of the abilities of NMDA agonists to displace the binding of [3H]CGP 39653 and [3H]Glu, but also caused diminution of those of NMDA antagonists. These results suggest that both sulfhydryl-reactive agents and phospholipases may predominantly interfere with radiolabeling of the NMDA recognition domain in a state favorable to an antagonist by [3H]CGP 39653, with concomitant facilitation of that in an agonist-preferring form by [3H]Glu. The possible presence of multiple forms of the NMDA recognition domain is further supported by these data.  相似文献   
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1α,25(OH)2-16-ene-D3, a synthetic analog of the steroid hormone, 1α,25(OH)2D3, has great potential to become a drug in the treatment of leukemia and other proliferative disorders, because of its minimal in vivo calcemic activity associated with a potent inhibitory effect on cell growth. However, at present, the mechanisms through which 1α,25(OH)2-16-ene-D3 expresses its biological activities are still not completely understood. Our previous in vitro study in a perfused rat kidney indicated for the first time that 1α,25(OH)2-16-ene-D3 and 1α,25(OH)2D3 are metabolized differently. 1α,25(OH)2-24-oxo-16-ene-D3, an intermediary metabolite of 1α,25(OH)2-16-ene-D3 formed through the C-24 oxidation pathway, accumulated significantly in the perfusate when compared to 1α,25(OH)2-24-oxo-D3, the corresponding intermediary metabolite of 1α,25(OH)2D3. In a subsequent in vivo study, we also reported that 1α,25(OH)2-24-oxo-16-ene-D3 exerted immunosuppressive activity equal to its parent, without causing significant hypercalcemia. In order to establish further the critical role of 1α,25(OH)2-24-oxo-16-ene-D3, in generating some of the key biological activities ascribed to its parent, we performed the present in vitro study using a human myeloid leukemic cell line (RWLeu-4) as a model. Comparative target tissue metabolism studies indicated that 1α,25(OH)2-16-ene-D3 and 1α,25(OH)2D3 are metabolized differently in RWLeu-4 cells, and the differences were similar to the ones we previously observed in the rat kidney. The significant finding was the accumulation of 1α,25(OH)2-24-oxo-16-ene-D3 in RWLeu-4 cells because of its resistance to further metabolism. Biological activity studies indicated that both 1α,25(OH)2-16-ene-D3 and its 24-oxo metabolite produced growth inhibition and promoted differentiation of RWLeu-4 cells to the same extent, and these activities were several fold higher than those exerted by 1α,25(OH)2D3. In addition, the genomic action of each vitamin D compound was assessed in a rat osteosarcoma cell line (ROS 17/2.8) by measuring its ability to transactivate a gene construct containing the vitamin D response element of the osteocalcin gene linked to the growth hormone reporter gene. In these studies, both 1α,25(OH)2-16-ene-D3 and its 24-oxo metabolite exerted similar but potent transactivation activity which was several fold greater than that exerted by 1α,25(OH)2D3 itself. In summary, our results indicate that the production and slow clearance of the bioactive intermediary metabolite, 1α,25(OH)2-24-oxo-16-ene-D3, in RWLeu-4 cells contributes significantly to the final expression of the enhanced biological activities ascribed to its parent analog, 1α,25(OH)2-16-ene-D3.  相似文献   
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报道了芦笋皮层组织培养再生植株的方法。表明:皮层在MS+1mg/L2,4—D+2mg/L6—BA培养基上形成无色或淡绿色疏松型愈伤组织,而MS+2mg/L6—BA+0.1mg/LIAA培养基则对芽的分化有利;试管苗的生很壮苗以无激素的MS为最佳培养基。此外,本试验还研究了不同碳源、不同激素组合对芦笋皮层离体培养再生植株的影响。  相似文献   
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中国种子植物区系定量化研究 V.区系相似性   总被引:3,自引:0,他引:3  
本文总结了应用相似系数即关联系数进行植物区系相似性分析的现状,指出了存在的问题.然后,从集合论角度讨论了区系相似性、相似关系及其相似系数的实质.作者以为在区系相似性分析中应用R.R.Sokal和C.D.Michener(1958)提出的简单匹配系数比较适宜,同时亦能避免以往区系相似性分析中缺乏可比性及某些“表相”相似等问题.最后,还提出了总体相似系数和类型相似系数二个新概念,以便按照吴征镒教授关于中国植物区系研究的学术思想统一研究各个不同地区植物区系的相似性.对此,作者用了6个区系实例进行了演算说明.  相似文献   
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利用EB病毒转化可产生较高水平人IgG和特异性抗2型登革病毒人抗体的Hu-TLC-SCID小鼠脾细胞,通过免疫组化、免疫荧光和PCR法检测转化细胞的人B细胞表面标志、EB病毒抗原和EB病毒基因。结果表明,被转化的Hu-TLC-SCID小鼠脾细胞能继续产生抗2型登革病毒的特异性人抗体,并具有人B细胞的、SmIgG标志及EB病毒潜伏膜蛋白-1(LMP-1)基因,可表达LMP-1和EB病毒核抗原(EBNA)。  相似文献   
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Lipid peroxide (LPO) in lymphocytes from mice was evaluated by measuring substances reactive to thiobarbituric acid (TBA). The product resulting from the reaction of TBA with lymphocytes was extracted with n-butyl and fluorescence intensity was determined. The degree of lipid peroxidation, expressed as fluorescence intensity f547, was assessed for stimulation of lymphocytes with concanavalin A (Con A), and was related to lymphocyte proliferation in response to Con A if Se was administered. The lymphocyte proliferation was determined by [3H]thymidine incorporation, expressed as cpm. The effect of superoxide dismutase (SOD), added to cell culture on lymphocyte proliferation was also evaluated. It was found that LPO in lymphocytes before Con A stimulation was significantly less than that after stimulation (p<0.001), and that SOD promoted lymphocyte proliferation dose dependently. The addition of Na2SeO3 to lymphocyte culture or supplementation in drinking water to mice decreased the produced LPO in lymphocyte in response to Con A. In the presence of Se, there is an inverse correlation between the levels of LPO in lymphocyte and the stimulated proliferation (r=−0.8902,r=−0.9439). In conclusion, active oxygen species scavenging was proposed as one of the mechanisms for Se to promote immunity.  相似文献   
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