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31.
J L Rubenstein J F Nicolas F Jacob 《Comptes rendus de l'Académie des sciences. Série III, Sciences de la vie》1984,299(8):271-274
We describe a general technique to inhibit gene expression in eukaryotic cells. The gene we chose to inhibit was the E. coli LacZ gene (encoding beta-galactosidase), which has previously been cloned into a eukaryotic expression vector [1]. This plasmid is called pCH110. We constructed a variant of pCH110 in which we flipped a 2566 base pair 5' fragment of the LacZ gene into the antiparallel orientation. The plasmid containing this mutated LacZ gene is called pNSLacZ (NS signifies non-sense coding sequence). When equal amounts of pCH110 and pNSLacZ are co-transfected into 3T6 mouse fibroblasts, the beta-galactosidase activity is decreased by approximately a factor of ten. Increasing the ratio of pNSLacZ to pCH110 above 1:1 does not appreciably increase the level of inhibition. Next, we prove the specificity of the inhibition by adding a third gene to the transfection mixture. For this purpose, we used pSVneo beta, a plasmid which expresses a phosphotransferase. We found that even when the beta-galactosidase activity was diminished by a factor of 10, the phosphotransferase activity was unaffected. Therefore, we have demonstrated that: the presence of an antiparallel copy of the LacZ gene results in a significant and specific diminution of the LacZ gene's expression; only a fraction of the LacZ gene needs to be in the antiparallel orientation in order to observe this effect. These results suggest that this technique can serve as a tool to decrease the level of gene expression in order to study the function of specific genes, or as a therapeutic manoeuvre in the treatment of disorders of abnormal gene expression. 相似文献
32.
Role of pancreatic enzymes in the intraluminal transport and ileal absorption of vitamin B 12 (Cb 1)
J P Nicolas G Marcoullis 《Comptes rendus des séances de la Société de biologie et de ses filiales》1980,174(4):548-563
The present paper outlines the classical concepts of transport and absorption of vitamin B12 and discusses findings which provide new insight into the important role of pancreatic enzymes in the absorption of the vitamin B12. In vivo experiments with healthy subjects and patients with exocrine pancreatic insufficiency demonstrate that the pancreatic enzymes do not activate "the precursor" intrinsic factor molecule but solely dissociate vitamin from the inactive R type proteins with a consequent coupling to the biologically active intrinsic factor. 相似文献
33.
Some diversity and niche amplitude parameters were applied to rangeland pastures of the Central Iberian Peninsula and to their succession stages after the periodical ploughing typical of the traditional management of these areas. Four different slopes within a large area of undulating terrain were selected for the monitoring of succession as they contained the characteristical geomorphological pattern of the area (denudation, transport and accumulation sectors).If we consider the total entropy theorem, H (E.P.)=H(E)+H(P/E), the total entropy of the slope H(E.P) and the entropy of species H(E) increase as succession progresses. As the value of the entropy of the sampling plots conditioned by the species H(P/E) is affected by the number of plots utilized, we employed the expression A=H(P/E)/log2 number of plots, similar to Pielou's index for niche amplitude, W=H(P/E)/H(P).This values decreases with succession, indicating that plant species tend to occupy more definite sectors along the slope. The number of low entropy species H(P/E)
i or specialist species, confined to narrow sectors also increases. When computed separately within the different sectors niche amplitude results in small values for the low slope regions (accumulation sector). This effect becomes more pronounced when succession advances.
Nous remercions le Conseil d'Administration de La Paranza, propriétaire du Castillo de Vañuelas, et particulièrement Mrs. C. Hernandez-Ros et J. A. Léon-Vrquijò, pour les faeilités qu'ils ont données à cette équipe durant la réalisation de ce travail. 相似文献
34.
J González-Cabrero R de Nicolas L Hernando J Egido 《Journal of immunology (Baltimore, Md. : 1950)》1991,147(12):4162-4166
We have recently described that patients with IgA nephropathy present high serum levels of anti-BSA idiotypic antibodies that were well correlated with the existence of hematuria. Furthermore, these Id were found in circulating and renal deposited immune complexes. In the present work, we examined the expression of surface idiotypic determinants on PBL by flow cytometry and their in vitro production, using as reagent anti-idiotypic antibodies previously well characterized. The presence of cross-reactive Id-bearing cells was observed in 5 out of 6 patients studied, with frequencies ranging from 3 to 12% of lymphocytes. After 7 days of culture, the spontaneous synthesis of idiotypic antibodies by PBL was found elevated in 6 out of 13 (46%) patients. A major Id cell expression and production was noted in patients with active disease as defined by hematuria. The preincubation of PBL with 20 and 50 micrograms of anti-idiotypic antibodies/2 x 10(6) cells for 3 days induced a significant inhibition of cross-reactive Id production in a dose-dependent fashion, with a degree of suppression between 12 and 50% in five out of six patients studied. In the above assays, as negative controls, we used the anti-Id antibodies previously adsorbed on an Id-Sepharose column. On the whole, these results suggest that patients with IgA nephropathy present dysfunctions in the Id-Anti-Id network that could play an important role in the pathogenesis of this disease. 相似文献
35.
36.
The developing mouse was used to assess biochemical changes in membrane lipids during the period when nerve growth cones become synapses. Growth cone particles and synaptosomes were simultaneously obtained from common brain homogenates. Incorporation of the essential fatty acid, docosahexaenoic acid (22:6 omega-3), was correlated with the developmental changes in endogenous fatty acid content of growth cones and synaptosomes. Analysis of endogenous lipid content indicated that, at all ages studied, the growth cones contained more arachidonoyl acyl chains (20:4 omega-6) than did synaptosomes. Before the onset of synaptogenesis, levels of arachidonoyl chains increased and levels of 22:6, oleoyl and linoleoyl chains decreased in synaptosomes. Although stearoyl and palmitoyl (16:0) remained stable in synaptosomes, 16:0 decreased in growth cones. With the exception of 16:0 and 20:4, endogenous fatty acyl content of growth cones and synaptosomes became similar by postnatal day 10, which coincides with the onset of synaptogenesis. When 5-day-old mouse pups were injected intraperitoneally with [3H]22:6, the incorporation into growth cone and synaptosome phospholipids was greatest in phosphatidylethanolamine, followed by phosphatidylserine and phosphatidylcholine. Nominal labeling was present in phosphatidic acid and phosphatidylinositol. Labeling in neutral lipids was less than that of phospholipids, with triacylglycerol incorporating most of the neutral lipid label, followed by diacylglycerol and free 22:6. Only the growth cone fraction contained detectable amounts of 22:6-labeled cholesterol esters. The distribution of 22:6 label in plasma 72 h after injection indicated that approximately 60% of the label was in phospholipids with approximately 40% in neutral lipids and less than 5% in free fatty acids.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
37.
Mohammed Sbia Marie-Francoise Diebler Nicolas Morel Maurice Israël 《Journal of neurochemistry》1992,59(4):1273-1279
The mediatophore is a presynaptic membrane protein that has been shown to translocate acetylcholine (ACh) under calcium stimulation when reconstituted into artificial membranes. The mediatophore subunit, a 15-kDa proteolipid, presents a very high sequence homology with the N,N'-dicyclohexylcarbodiimide (DCCD)-binding proteolipid subunit of the vacuolar-type H(+)-ATPase. This prompted us to study the effect of DCCD, a potent blocker of proton translocation, on calcium-dependent ACh release. The present work shows that DCCD has no effect on ACh translocation either from Torpedo synaptosomes or from proteoliposomes reconstituted with purified mediatophore. However, using [14C]DCCD, we were able to demonstrate that the drug does bind to the 15-kDa proteolipid subunit of the mediatophore. These results suggest that although the 15-kDa proteolipid subunits of the mediatophore and the vacuolar H(+)-ATPase may be identical, different domains of these proteins are involved in proton translocation and calcium-dependent ACh release and that the two proteins have a different membrane organization. 相似文献
38.
39.
Jean- -François Laliberté Olivier Nicolas Serge Durand Rolf Morosoli 《Plant molecular biology》1992,18(3):447-451
The xylanase gene from Cryptococcus albidus contains seven introns. Genomic and cDNA clones under the control of the CaMV 35S promoter were transferred into tobacco plants using Agrobacterium-mediated cell transformation. The genes were transcribed and the mRNAs were amplified by the polymerase chain reaction using primers on each side of the intron region. About 90% of the amplification products from plants transformed with the genomic clone corresponded to the size of the pre-mRNA (1.2 kb) and 10% represented the spliced product (0.85 kb). The 0.85 kb fragment was cloned and sequenced and the result indicated that the introns from the xylanase gene were accurately spliced by the plant cells. 相似文献
40.
Conclusion From this brief review it appears that at least three categories of human glioma-associated antigens may exist. The first seems to be restricted and common to gliomas. The second is shared between gliomas, normal adult brain, and fetal brain. The third is present on cells from adult and fetal tissue and on cells from tumours derived from the neural crest. The expression of glioma-associated antigens is highly variable from one tumour, or tumour cell line, to another, and reflects the phenotypic heterogeneity of the glioma group. Moreover, this heterogeneity has been found in different clones of individual glioma cell lines [1]. The fact that gliomas share some antigens with normal brain is of critical importance for immunodiagnosis or immunotherapy. It is evident that active immunotherapy for gliomas should be performed with cultured cells and not with tumour extracts, because such extracts may contain MBP.The exact nature of the various glioma-associated antigens remains to be clearly defined, however. They may belong to a group of surface glycoproteins such as those described by Lloyd et al. [24] for melanoma or more recently by Lubitz et al. [25] for glial cells. 相似文献