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111.
Denis Tagu Catherine Bergounioux Claudette Perennes Pierre Gadal 《Plant Cell, Tissue and Organ Culture》1990,21(3):259-266
In previous work, transformedPetunia hybrida plants were obtained by direct gene transfer, using two different genes on separate plasmids (NPT II gene and a cDNA of PEPC from green sorghum leaves). In this study, we have analysed the sexual transmission of the acquired genes by genetic crossing analysis of 2 of the transgenic petunias. The ploïdies of the two clones were determined by flow cytometric analysis showing that one was 2n and the other 4n. Self and back crosses show that the kanamycin character was inherited as a single dominant trait, and that the two clones were heterozygotes for this character. Therefore, the 4n clone probably arises from an endoploidization followed by a transformation event. Southern blot analyses show that all of the resistant progenies which were analysed harboured the kanamycin gene, and expressed the phosphorylation activity in vitro. The DNA of several progenies were also tested for the presence of co-transformed PEPC cDNA sequence. All of the kanamycin-resistant progenies tested contained the second coding sequence, indicating that the two foreign genes might be genetically co-inherited in the transgenic plants. The way in which the two genes are integrated into the genome is discussed.Abbreviations NPT
neomycin phosphotransferase
- PEG
polyethyleneglycol
- PEPC
phosphoenolpyruvate carboxylase 相似文献
112.
Pascale Andre Christian Capo Anne Marie Benoliel Michel Buferne Pierre Bongrand 《Cell biochemistry and biophysics》1990,16(1-2):13-34
Fluorescent probes are widely used to study cell structure and function. However, few reports were devoted to a quantitative analysis of the intracellular distribution of fluorescent markers. In the present work, we describe the topographical changes of surface and cytoskeletal markers on individual cells subjected to adhesive or mechanical interaction. Conjugates were prepared with a cytotoxic T-lymphocyte clone and target cells. Specific antigens, membrane phospholipids, surface glycoconjugates, and polymerized actin were labeled with fluorescent antibodies or biochemical probes. The analysis of fluorescence distributions in conjugates demonstrated a selective reorganization of the plasma membrane with a gathering of some molecular species in the intercellular adhesion area. Furthermore, individual phagocytic cells were sucked into glass micropipets, then stained with fluorescent phallacidin to analyze the effect of mechanical efforts on the cytoskeleton organization. The concentration of polymerized actin was found to be similar in mechanicallyinduced protrusions and whole cells. It is concluded that adhesive interactions may result in marked cell polarization and formation of membrane zones with a particular biochemical composition. The submembranar cytoskeleton might play a role in this process. 相似文献
113.
Marianne Horoyan Anne-Marie Benoliel Christian Capo Pierre Bongrand 《Cell biochemistry and biophysics》1990,17(3):243-256
We combined fluorescence labeling, digital image processing, and micromanipulation to investigate the intracellular events induced by inflicting a mechanical stress on rat basophilic leukemia cells. Our findings were as follows:
- Most cells displayed a localized calcium rise in response to micropipet aspiration. This represented an average threefold increase as compared to resting level, and it was observed during the first 10 s following aspiration. A slow return to initial level occurred within about 3 min. Further, this calcium rise involved a mobilization of intracellular stores, since it was not prevented by adding a calcium chelator into the extracellular medium.
- All micropipet-aspirated cells displayed a local accumulation of microfilaments, with a preferential localization in the cell protrusions or near the pipet tips.
- No absolute correlation was found between the localization of calcium rise and cytoskeletal accumulation.
- Cell deformability was decreased when intracellular calcium was maintained at a constant (high or low) level with ionomycin and/or EGTA.
114.
Pierre Tachon 《Free radical research》1990,9(1):39-47
The role of histidine on DNA breakage induced by hydrogen peroxide (H2O2) and ferric ions or by H2O2 and cupric ions was studied on purified DNA. L-histidine slightly reduced DNA breakage by H2O2 and Fe3+ but greatly inhibited DNA breakage by H2O2 and Cu2+. However, only when histidine was present, the addition of EDTA to H2O2 and Fe3+ exhibited a bimodal dose response curve depending on the chelator metal ratio. The enhancing effect of histidine on the rate of DNA degradation by H2O2 was maximal at a chelator metal ratio between 0.2 and 0.5, and was specific for iron. When D-histidine replaced L-histidine, the same pattern of EDTA dose response curve was observed. Superoxide dismutase greatly inhibited the rate of DNA degradation induced by H2O2, Fe3+, EDTA and L-histidine involving the superoxide radical.
These studies suggest that the enhancing effect of histidine on the rate of DNA degradation by H2O2 and Fe3+ is mediated by an oxidant which could be a ferrous-dioxygen-ferric chelate complex or a chelate-ferryl ion. 相似文献
These studies suggest that the enhancing effect of histidine on the rate of DNA degradation by H2O2 and Fe3+ is mediated by an oxidant which could be a ferrous-dioxygen-ferric chelate complex or a chelate-ferryl ion. 相似文献
115.
116.
From 1987 until 1995, life-history traits of the Lesser Sheathbill Chionis minor in the Kerguelen Archipelago were compared with those at other localities, where this species breeds in seabird colonies. At Kerguelen Island only, some pairs breed on shores free of penguin and cormorant colonies. Moreover, pairs, including nonbreeders, and solitary individuals maintain territories all the year round. Site and mate changes were not linked, and all divorces were permanent. Fidelity rates were similar in all localities and so were the annual survival rates of adults. The survival rate of immature individuals was highest at Kerguelen Island, where sheathbills laid smaller clutches, produced fewer fledglings and had lower breeding success than in the other islands. At Kerguelen Island, pairs breeding on shores had similar clutch sizes but fledged fewer offspring than those breeding in seabird colonies. Further, the proportion of vacant territories and nonbreeders was higher on shores. Kerguelen sheathbills devoted less time to food acquisition than those on Marion Island, being exceptionally kleptoparasitic, and spent a smaller part of their foraging time exploiting seabird colonies. Their diet was mainly algae. The differences on Kerguelen Island as compared with other localities were caused by the presence of an extensive intertidal zone on the former, which reduced competition, making many more sites suitable and the environment more predictable. These life-history and behavioural traits are discussed in relation to life-history theory. As differences were also found, although less extensive, within the Kerguelen Archipelago, we suggest that some traits represent an adaptive response to external constraints and that the life-history strategy of the Lesser Sheathbill is particularly opportunistic. 相似文献
117.
Olivier Cohen Christine Cans Jean Louis Gilardi Hubert Roth Marie-Ange Mermet Pierre Jalbert Jacques Demongeot Martine Cuillel 《Human genetics》1996,97(5):659-667
Reciprocal translocations (rcp) are among the most common constitutional chromosomal aberrations in man. Using a European
database of 1574 families carrying autosomal rcp, a cartographic study was done on the breakpoints involved. The breakpoints
are non-randomly distributed along the different chromosomes, indicating “hot spots”. Breakpoints of rcp that result in descendants
that are unbalanced chromosomally at birth are more frequent in a distal position on chromosomal arms, and 65% of them are
localised in R-bands. Among the R-bands, bands rich in GC islands and poor in Alu repetitive sequences are more frequently
the site of breakpoints, as well as bands that include a fragile site. This result suggests that the variation in degree of
methylation in GC islands could be involved in chromosomal breakage and hence in chromosomal rearrangements.
Received: 10 April 1995 / Revised: 1 July 1995 相似文献
118.
Maria de Fatima Bonaldo Pierre Jelenc Long Su Lee Lawton M. -T. Wu Dorothy Warburton Marcelo Bento Soares 《Human genetics》1996,97(4):441-452
A study was conducted on the feasibility of isolating genes and pseudogenes that map to chromosome 13 by a hybridization-based approach using a 13-specific library and pools of repeat-free cDNA clones. Five pairs of cDNA and chromosome 13 genomic clones were identified and characterized. Partial or full-length sequence was derived from all cDNAs, and database searches were performed for putative gene identification. Partial sequence was also obtained from the chromosome 13 genomic clones for comparison with those of the hybridizing cDNAs. As a result of these analyses we identified three genes, a putative homologue of a porcine mRNA encoding an unidentified hepatic protein, a putative homologue of a yeast integral membrane protein, and a gene for a translationally controlled tumor protein, and two processed pseudogenes, ribosomal proteins L23a and S3a. The latter was formerly identified as the v-fos transformation effector gene, Fte-1, and recently cited as a possible candidate for the BRCA2 gene on chromosome 13. All genes and pseudogenes were localized to cytogenetic bands by in situ hybridization of metaphase chromosomes with probes derived from the chromosome 13 genomic clones. 相似文献
119.
We review some of the older literature concerning metabolic turnover of cholesterol in the nervous system. The overall picture
is that incorporation of radioactive precursors into brain cholesterol is roughly proportional to the rate of myelination
and that, once incorporated, radioactive cholesterol is relatively stable metabolically. We outline a strategy for demonstrating
the source (local synthesis or uptake from the circulation) of cholesterol in brain. The experimental design involves determining
the rate of accumulation of cholesterol this is calculated as the increasing amounts of sterol in brain at successive time
intervals during development. The rate of appearance of newly synthesized cholesterol is determined from incorporation of
radioactivity from3H2O (injected i.p. several hours prior to sacrifice) into cholesterol. The radioactivity associated with the sterol fractions
and the specific activity of body water determined from the serum can be used to calculate the absolute amount of sterol newly
synthesized during the time when3H2O was present. The results obtained demonstrated that all of the bulk cholesterol accumulating in brain can be accounted for
by newly synthesized cholesterol. None of the radioactive cholesterol came from the circulation, since cholesterol feeding
suppressed cholesterol biosynthesis in the liver and specific radioactivity of circulating cholesterol was negligible. Thus,
almost all cholesterol accumulating in brain during development is locally synthesized.
Special issue dedicated to Dr. Marion R. Smith. 相似文献
120.
Josiane Arnaud Pierre Bourlard Bernard Denis Alain E. Favier 《Biological trace element research》1996,53(1-3):129-136
This study was carried out to assess manganese (Mn) status after an acute episode of myocardial infarction. Plasma and erythrocyte
Mn concentrations were measured from admission to hospital to day 15 postadmission in 21 patients suffering from acute myocardial
infarction and in three control groups. The determination of Mn in these biological fluids was performed by electrothermal
atomic absorption spectrometry. Plasma Mn was higher (p<0.01) and erythrocyte Mn was similar in the acute myocardial infarction group compared to healthy age-matched control group.
Plasma and erythrocyte Mn remained unchanged during the 2 wk after acute myocardial infarction and were not correlated to
enzyme activities. A decrease of erythrocyte Mn with age, expressed in nmol/L, was noted (p<0.02). These results suggest that plasma and erythrocyte Mn do not provide an indication of myocardial damage. Nonetheless,
Mn status in elderly merits further attention. 相似文献