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71.
Biochemical implications of sequence comparisons of the cystic fibrosis transmembrane conductance regulator 总被引:2,自引:0,他引:2
System A, the Na(+)-dependent amino acid transport activity, is encoded by the ATA2 gene and up-regulated following partial hepatectomy (PH), and its competitive inhibition interferes with liver regeneration. Rabbit polyclonal antibody was raised against a portion of the ATA2 gene product followed by immunodetection of ATA2 in isolated liver plasma membrane and lysate. The level of ATA2 increased in the plasma membrane following PH, while the relatively high quantity of ATA2 found in liver lysate remained constant. We also have shown that Northern analysis of steady-state ATA2 mRNA revealed no significant change following PH. These data show that ATA2-mediated transport is not regulated by the steady-state level of ATA2 mRNA but is regulated by the amount of ATA2 and redistribution to the plasma membrane. We hypothesize that ATA2 activity is regulated by recruitment of ATA2 protein from an intracellular compartment. In addition, the pattern of expression of System A activity in oocytes, transport kinetics, and sensitivity to chemical modification indicate the presence of a second System A isoform in liver that differs substantially from ATA2. 相似文献
72.
Two types of males are present in the Jalapa population of Xiphophorus helleri: Males with a black or dark brownish mid-lateral stripe ("black males") mature earlier and are smaller than males with a red or brownish mid-lateral stripe ("red males"). We tested the hypothesis that the colour patterns of red males may be regarded as ornaments, which evolved as the result of inter- and/or intrasexual selection. As predicted, in choice tests females exhibited a strong preference for red versus black males. Furthermore, in competition experiments red males became dominant over black males with no exception, both when body size was equal and even when red males were 3–5 mm smaller than black males. However, contrary to our prediction, sneak–chase behaviour in black males was not detected, and courtship displays occurred at similar rates in both morphs. Red males are always heterozygous, with one allele for red and another for black colouration. This genetic constraint potentially prevents the extinction of the black morph. Possibly, the colour patterns of red males are functioning as indicators of heterozygosity. According to the "heterozygosity theory of mate choice", the female's preference for red males may be adaptive. 相似文献
73.
A polarized cell: the root statocyte 总被引:2,自引:0,他引:2
In the gravity-perceiving cells (statocytes), located in the centre of the root cap, polarity is expressed in the arrangement of the organelles since, in most genera, the nucleus and the endoplasmic reticulum are maintained at the opposite ends of each cell by actin. Polarity is also evident in the distribution of plasmodesmata, which are more numerous in the transverse walls than in the longitudinal walls. The centre of each statocyte is depleted of microtubules (they are only located at the periphery) but is occupied by numerous amyloplasts (statoliths), denser than the cytoplasm. The amyloplasts do not contribute to the inherent structural polarity since their position is dependent upon the gravity vector. This article focuses on new microscopic analyses and on data obtained from experiments performed in microgravity, which have contributed to our better understanding of the architecture of the actin web implicated in the perception of gravity. Depending upon the plant, the actin network seems to be formed of single filaments arranged in various ways, or, of thin bundles of actin filaments. The amyloplasts are enmeshed in this web of actin and their envelopes are associated with it, but they can have autonomous movement via myosin in the absence of gravity. From calculations of the value of the force necessary to move one amyloplast in the lentil root, and from videomicroscopy performed with living statocytes of maize roots, it is hypothesized that actin microfilaments could be orientated in an overall diagonal direction in the statocyte. These observations could help in understanding how slight amyloplast movements may trigger and transmit the gravitropic signal. 相似文献
74.
Novel derivatives of 9,10-anthraquinone are selective algicides against the musty-odor cyanobacterium Oscillatoria perornata 总被引:2,自引:0,他引:2
Schrader KK Nanayakkara NP Tucker CS Rimando AM Ganzera M Schaneberg BT 《Applied and environmental microbiology》2003,69(9):5319-5327
Musty "off-flavor" in pond-cultured channel catfish (Ictalurus punctatus) costs the catfish production industry in the United States at least 30 million US dollars annually. The cyanobacterium Oscillatoria perornata (Skuja) is credited with being the major cause of musty off-flavor in farm-raised catfish in Mississippi. The herbicides diuron and copper sulfate, currently used by catfish producers as algicides to help mitigate musty off-flavor problems, have several drawbacks, including broad-spectrum toxicity towards the entire phytoplankton community that can lead to water quality deterioration and subsequent fish death. By use of microtiter plate bioassays, a novel group of compounds derived from the natural compound 9,10-anthraquinone have been found to be much more selectively toxic towards O. perornata than diuron and copper sulfate. In efficacy studies using limnocorrals placed in catfish production ponds, application rates of 0.3 micro M (125 micro g/liter) of the most promising anthraquinone derivative, 2-[methylamino-N-(1'-methylethyl)]-9,10-anthraquinone monophosphate (anthraquinone-59), dramatically reduced the abundance of O. perornata and levels of 2-methylisoborneol, the musty compound produced by O. perornata. The abundance of green algae and diatoms increased dramatically 2 days after application of a 0.3 micro M concentration of anthraquinone-59 to pond water within the limnocorrals. The half-life of anthraquinone-59 in pond water was determined to be 19 h, making it much less persistent than diuron. Anthraquinone-59 appears to be promising for use as a selective algicide in catfish aquaculture. 相似文献
75.
Ostafin AE Ponomarenko NS Popova JA Jäger M Bylina EJ Norris JR 《Photosynthesis research》2003,77(1):53-68
The utility of photosynthetically defective mutants in the purple photosynthetic bacterium Blastochloris viridis (formerly Rhodopseudomonas viridis)was demonstrated with construction of a reaction-center deficient mutant, LH 1-H. This LH 1-H mutant has a photosynthetic
apparatus in which most of the puf operon genes were deleted, resulting in an organism containing only the genes for the light harvesting polypeptides and the
H subunit of the reaction center. This B. viridisstrain containing a truncation of the puf operon was characterized by gel electrophoresis, lipid-to-protein ratio analysis, optical spectroscopy, electron paramagnetic
resonance and transmission electron microscopy. Optical and electron paramagnetic resonance spectroscopies revealed no photoactivity
in this LH 1-H mutant consistent with the absence of intact reaction centers. Electron paramagnetic resonance evidence for
assembled LH 1 complexes suggested that the interactions between light harvesting polypeptide complexes in membranes were
largely unchanged despite the absence of their companion reaction center cores. The observed increase in the lipid-to-protein
ratio was consistent with modified interactions between LH 1s, a view supported by transmission electron microscopy analysis
of membrane fragments. The results show that B. viridis can serve as a practical system for investigating structure-function relationships in membranes and photosynthesis through
the construction of photosynthetically defective mutants.
This revised version was published online in August 2006 with corrections to the Cover Date. 相似文献
76.
A novel method for the covalent attachment of erythrocytes to glass microscope coverslips that can be used to image intact
cells and the cytoplasmic side of the cell membrane with either solid or liquid mode atomic force microscopy (AFM) is described.
The strong binding of cells to the glass surface is achieved by the interaction of cell membrane carbohydrates to lectin,
which is bound to N-5-azido-2-nitrobenzoyloxysuccinimide (ANBNOS)-coated coverslips (1). The effectiveness of this method
is compared with the other commonly used methods of immobilizing intact erythrocytes on glass coverslips for AFM observations.
Experimental conditions of AFM imaging of biologic tissue are discussed, and typical topographies of the extracellular and
the cytoplasmic surfaces of the plasma membrane in the dry state and in the liquid state are presented. Comparison of the
spectrin network of cell age-separated erythrocytes has demonstrated significant loss in the network order in older erythrocytes.
The changes are quantitatively described using the pixel height histogram and window size grain analysis. 相似文献
77.
78.
The effect of mannitol on different genotypes of potato was studied in callus and plantlet culture. In vitro responses of five potato genotypes with well-known field behaviour to water deficit were analysed. After a 4-week-long cultivation on media containing mannitol up to 0.8 M, different morpho-physiological parameters were determined and statistically analysed. The useful concentration of mannitol for in vitro screening the osmotic tolerance of different genotypes depended on the type of culture; it was 0.4 M in plantlet-test and 0.8 M in callus-test. In callus-test the relative increase of callus mass was a useful parameter for determination of osmotic tolerance of genotypes at cellular level. In plantlet culture, stress index calculated from the rate of surviving in vitro shoots, number and length of roots per surviving explant and the rate of rooted explants were applicable to determine three groups according to the tolerant, medium tolerant and sensitive categories in agreement with the field behaviour of these genotypes. Under in vitro stress conditions we were able to distinguish the examined genotypes with different drought tolerance. 相似文献
79.
The reaction between low molecular weight S-nitrosothiols and hemoglobin is often used to synthesize S-nitrosohemoglobin, a form of hemoglobin suggested to be involved in the regulation of vascular oxygen delivery. However, this reaction has not been studied in detail, and several groups have reported a variable co-formation of oxidized methemoglobin (metHb) during synthesis. This study examines the mechanism of metHb formation and shows that nitrosylhemoglobin (HbNO) can also be formed. Generation of metHb and HbNO is largely dependent on the presence of protein thiol groups. We present evidence for a mechanism for the formation of metHb and HbNO involving the intermediacy of nitroxyl anion. Specifically, the reaction of nitroxyl with S-nitrosothiols to liberate nitric oxide and reduced thiol is proposed to be central to the reaction mechanism. 相似文献
80.
Alicia I. Cardoso Nathalie Sixt Agnes Vallier Joel Fayolle Robin Buckland T. Fabian Wild 《Journal of virology》1998,72(3):2516-2518
Plasmids encoding the measles virus hemagglutinin (HA) and nucleoprotein (NP) proteins inoculated into the skin of BALB/c mice by the gene gun method induced both humoral and cytotoxic lymphocyte class I-restrict- ed immune responses. Although intramuscular immunization induces the immunoglobulin G2a (IgG2a) antibody isotype for both antigens, with gene gun immunization, the NP still generated mainly IgG2a and the major isotype induced by the HA was IgG1. Interestingly, gene gun coimmunization of HA and NP plasmids resulted in a dominant IgG1 HA response and the switching of antibodies generated against the NP to the IgG1 isotype.The initial studies showing that injection of DNA into muscle induces an immune response to the encoded protein opened a new approach to vaccination (for reviews, see references 19 and 22). Recent studies suggest that inoculated muscle cells probably act only as a source of antigen and that immune priming takes place elsewhere in the body (14). For example, excision of an injected muscle a few minutes after DNA inoculation did not affect antibody and cytotoxic T-lymphocyte (CTL) responses (21). Thus, it may be interesting to examine other DNA delivery systems to study how the immune system responds to DNA vaccination. One alternative system involves precipitating DNA onto gold beads which are then propelled into the skin by means of pressurized helium gas (12). When such a system is used, less DNA is required, but unlike the case with intramuscular inoculations, the response is Th2-like, generating immunoglobulin G1 (IgG1) antibodies (17). More recent observations suggest that this is probably due to the mode of inoculation rather than the route (10). We have been studying DNA vaccination against the paramyxovirus measles virus (MV). This disease is one of the primary causes of infant mortality in developing countries, and there is an urgent need for an effective vaccine in infants, as the present live attenuated vaccine is inefficient in the presence of maternal antibodies. Our previous studies established that in a mouse model at least three MV proteins play a role in protection (23). Both glycoproteins, hemagglutinin (HA) and fusion, induce neutralizing antibodies (9, 11), and HA and nucleoprotein (NP) induce CTLs (3, 4), which do not protect against infection but help in recovery (5). In our previous study on DNA vaccination, we showed that intramuscular inoculation of DNAs coding for the MV HA and NP (pV1J-HA and pV1J-NP [6]) induced class I-restricted CTLs and a humoral response corresponding to a Th1 response (6). In the present study, we have extended our observations to compare the same plasmids’ ability to induce an immune response when they are delivered into the skin by a gene gun (Bio-Rad, Ivry sur Seine, France). Gold beads were coated with DNA as follows: approximately 30 mg of gold powder (1.0-μm gold beads; Bio-Rad) was mixed with 100 μl of 0.1 M spermidine (Sigma, L’Isle D’Abeau, France). After sonication, 0.5, 2, or 5 μg of plasmid DNA was added per mg of gold powder, and then 200 μl of 2.5 M CaCl2 was added to the mixture, with gentle vortexing. Pellets were washed three times and suspended in cold 100% ethanol. Tubes containing dried DNA-coated gold beads were stored at 4°C.