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Determination of the chirality of amino acid residues in the course of subtractive Edman degradation of peptides. 总被引:1,自引:0,他引:1
A chiral reagent, 1-fluoro-2,4-dinitro-5-L-alanine, was synthesized for the analysis of enantiomeric mixtures of amino acids after precolumn derivatization. The resulting diastereomers can be separated and quantitated by microbore RP-HPLC. These derivatives are relatively stable under the conditions used for acid hydrolysis of peptide bonds. Thus, this reagent was included in the protocol of a subtractive Edman degradation procedure of peptides to determine the sequence position of amino acid residues with concomitant identification of their chirality at a nanomolar level. 相似文献
24.
Maurizio Rossetto Kingsley W. Dixon Eric Bunn 《In vitro cellular & developmental biology. Plant》1992,28(4):192-196
Summary Aeration of tissue cultured rare Australian plantsConostylis wonganensis S.D. Hopper (Haemodoraceae);Diplolaena andrewsii Ostenf.;Drummondita ericoides Harvey (Rutaceae);Eremophila resinosa F. Muell. (Myoporaceae);Eucalyptus ‘graniticola’ (Myrtaceae);Lechenaultia pulvinaris C. Gardner (goodeniaceae); andSowerbaea multicaulis E. Pritzel (Liliaceae) has been found to reduce vitrification in sensitive species as well as significantly improving shoot
quality and transfer to soil in most study species. A simple 7-mm hole with a double-layer insert of filter paper in the polypropylene
screw lids of the culture vessel decreased shoot vitrification over a 4-wk culture period. The method has implications for
facilitating the tissue culture of other rare Australian plants and reducing the occurrence of this developmental abnormality. 相似文献
25.
Purification and characterization of bioactive peptides from skin extracts of Rana esculenta 总被引:2,自引:0,他引:2
M Simmaco D De Biase C Severini M Aita G F Erspamer D Barra F Bossa 《Biochimica et biophysica acta》1990,1033(3):318-323
The peptide fraction extracted by methanol from the skin of Rana esculenta, a species widely distributed in Western Europe, was investigated. The pharmacological activity found in the extract is attributable to the presence of authentic bradykinin, together with a shorter, partially active version of this molecule, des-Arg9-bradykinin. Also the bradykinin fragment 1-7 has been isolated, but it was inactive in our bioassay system. Moreover, a family of hydrophobic peptides has been purified and characterized, which appeared devoid of pharmacological activities when tested on smooth muscle preparations, but were provided with hemolytic activities. 相似文献
26.
27.
Giuseppe Fossati Andrea Balsari Donatella Taramelli Maria Luisa Sensi Giuseppe Pellegris Maurizio Nava Giorgio Parmiani 《Cancer immunology, immunotherapy : CII》1982,14(2):99-104
Summary Peripheral blood lymphocytes (PBL) of melanoma patients were sensitized in vitro with lymphocytes of a single donor or with a pool of lymphocytes of 5–20 different donors. After 6–7 days, the cytotoxic activity of the sensitized PBL was tested against cultured autologous tumor cells and lymphocytes in a 51Cr-release assay. Tumor lysis was observed in 13 of 16 cases in which patients' PBL (Pt-PBL) were stimulated by a pool of allogeneic lymphocytes and in five out of seven cases when single sensitization was performed. In no case was lysis of autologous normal lymphocytes or blasts seen. Cultivation of Pt-PBL with irradiated autologous tumor cells never led to the induction of lymphocytes cytotoxic to melanoma cells. Lysability by pool-activated autologous Pt-PBL of fresh cryopreserved tumor cells was compared to that of short-term cultured tumor cells, and no significant differences were observed. Cold-target inhibition experiments indicated that the cytotoxicity of Pt-PBL was tumor-restricted since only autologous melanoma cells but not lymphocytes were able to inhibit the reaction. These results indicate that activation of Pt-PBL is necessary in order to elicit or amplify their antitumor activity. 相似文献
28.
George W. Arana Ross J. Baldessarini Michael Herschel Maurizio Fava 《Life sciences》1981,29(2):121-133
The binding of low concentrations of [3H](?)apomorphine to preparations of calf and rat forebrain tissue was evaluated. Fractionation of crude homogenates to prepare a membrane fraction (P4) of striatal or caudate homogenates increased the proportion of saturable to total binding from 33% to over 80%, and increased the apparent density of binding sites from 94 to 681 fmol/mg protein. Binding in calf caudate P4 tissue was protein-dependent and optimal at pH = 7.0 to 7.5, and T = 20 to 25°C; at higher temperatures tissue binding sites appeared to degrade. The half-time of association and dissociation at 22°C were, respectively, 14.0 and 18.5 min; equilibration was complete in 60 min. Kinetic characteristics of high-affinity binding obtained from association and dissociation constants and from saturation isotherms were similar (Kd = 2.1 to 3.4 nM). The pharmacology of competition for 3H-APO suggests selectivity for dopamine-agonist interactions. These results indicate that the P4 membrane preparation may be useful for the evaluation of dopamine-agonist binding sites or “receptors.” 相似文献
29.
Cutolo M Capellino S Montagna P Ghiorzo P Sulli A Villaggio B 《Arthritis research & therapy》2005,7(5):R1124-R1132
Sex hormones seem to modulate the immune/inflammatory responses by different mechanisms in female and male rheumatoid arthritis
patients. The effects of 17β-oestradiol and of testosterone were tested on the cultured human monocytic/macrophage cell line
(THP-1) activated with IFN-γ in order to investigate their role in cell proliferation and apoptosis. Activated human THP-1
cells were cultured in the presence of 17β-oestradiol and testosterone (final concentration, 10 nM). The evaluation of markers
of cell proliferation included the NF-κB DNA-binding assay, the NF-κB inhibition complex, the proliferating cell nuclear antigen
expression and the methyl-tetrazolium salt test. Apoptosis was detected by the annexin V-propidium assay and by the cleaved
poly-ADP ribose polymerase expression. Specific methods included flow analysis cytometry scatter analysis, immunocytochemistry
and western blot analysis. Cell growth inhibition and increased apoptosis were observed in testosterone-treated THP-1 cells.
Increased poly-ADP ribose polymerase-cleaved expression and decreased proliferating cell nuclear antigen expression, as well
as an increase of IκB-α and a decrease of the IκB-α phosphorylated form (ser 32), were found in testosterone-treated THP-1
cells. However, the NF-κB DNA binding was found increased in 17β-oestradiol-treated THP-1 cells. The treatment with staurosporine
(enhancer of apoptosis) induced decreased NF-κB DNA binding in all conditions, but particularly in testosterone-treated THP-1
cells. Treatment of THP-1 by sex hormones was found to influence cell proliferation and apoptosis. Androgens were found to
increase the apoptosis, and oestrogens showed a protective trend on cell death – both acting as modulators of the NF-κB complex. 相似文献
30.
Maurizio Lambardi Kiran K. Sharma Trevor A. Thorpe 《In vitro cellular & developmental biology. Plant》1993,29(4):189-199
Summary Studies were undertaken to optimize tissue culture conditions for micropropagation of Aleppo pine (Pinus halepensis Mill.) from mature embryos and various explants of the embryo. Over 90% of the embryo explants gave rise to adventitious
buds within 4 wk. Intact embryos were the most suitable explants for shoot bud induction. Both isolated cotyledons and hypocotyls
produced adventitious buds, but these developed slowly and failed to elongate. N6-Benzyladenine (BA) alone at 5.0μM was the most effective cytokinin when added to gelled to gelled von Arnold and Eriksson’s (AE) medium containing 3% sucrose.
Adventitious bud development was achieved on hormone-free AE medium, and shoot elongation was optimum on three quarter-strength
Bornman’s MCM medium, with 0.1% conifer-derived activated charcoal. Shoots were multiplied on three-quarter strength MCM medium,
containing 5μM BA. To induce adventitious roots on the elongated shoots, pulse treatment with 1 mM IBA for 6 h, followed by the transfer of the shoots to sterile peat:vermiculite (1:1) mixture, was beneficial. After acclimatization
for 3 to 4 wk under mist, almost all the rooted shoots could be transplanted successfully to the greenhouse, where the plants
exhibited normal growth habit. Histologic studies on the ontogeny of adventitious shoot formation from mature embryo explants
revealed temporal structural changes in different parts of the explant. Induction of mitotic divisions on the shoot-forming
medium resulted in the formation of meristemoids in the epidermal and subepidermal layers of the explant, located initially
at both the tips of the cotyledons and the axils of adjacent cotyledons. Shoot buds arising in the axils of adjacent cotyledons
were due to new cell division and not to any preexisting meristem. 相似文献