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71.
72.
Coppoolse ER de Vroomen MJ van Gennip F Hersmus BJ van Haaren MJ 《Plant molecular biology》2005,58(5):687-698
Cre/lox recombination in vivo has become an important tool to induce chromosomal rearrangements like deletions. Using a combination of Ds transposition and Cre/lox recombination in two independent experiments on chromosomes 6 and 7 of tomato, two sets of somatic deletions up to a size of 200 kb were obtained. The efficiency of somatic deletion decreased with increasing deletion size. The largest germinally transmitted deletion had a size of only 55 kb. The results show that Cre-mediated deletion in somatic cells is less efficient when the lox sites are separated over larger distances. A further drop of the deletion efficiency after germinal transmission of the larger deletions can be explained by the probable loss of genes that are of vital importance to gametophyte function. Plasmid rescue of an 8.4 kb circularised deleted DNA showed that the Cre-mediated deletion takes place in tomato as expected. Since the circular Cre-deleted DNA could only be PCR amplified in plant cells where the deletion was not complete, the double-stranded DNA circle is assumed to be instable. 相似文献
73.
Mutational study of the bacterial hemoglobin distal heme pocket 总被引:1,自引:0,他引:1
Verma S Patel S Kaur R Chung YT Duk BT Dikshit KL Stark BC Webster DA 《Biochemical and biophysical research communications》2005,326(2):290-297
Ligand binding experiments on three mutants in the distal heme pocket of Vitreoscilla hemoglobin (GlnE7His, ProE8Ala, and GlnE7His,ProE8Ala) were used to probe the role of GlnE7 and ProE8 in the pocket's unusual structure. The oxygen dissociation constants for the wild type, E8Ala mutant, and E7His mutant proteins were 4.5, 4.7, and 1.7microM, respectively; the K(d) for the double mutant was not determinable by our technique. Visible-Soret spectra of the carbonyl and cyanyl forms and FT-IR of the carbonyl form of the E8 mutant were similar to those of the wild type; the opposite was true for the GlnE7His and GlnE7His,ProE8Ala mutants, which also differed from wild type in the visible-Soret spectra of their oxidized forms. Models of the effects of the mutations on distal pocket structure were consistent with the experimental findings, particularly the larger effects of the GlnE7His change. 相似文献
74.
Klonowska A Clark ME Thieman SB Giles BJ Wall JD Fields MW 《Applied microbiology and biotechnology》2008,78(6):1007-1016
Desulfovibrio vulgaris Hildenborough is a well-studied sulfate reducer that can reduce heavy metals and radionuclides [e.g., Cr(VI) and U(VI)].
Cultures grown in a defined medium had a lag period of approximately 30 h when exposed to 0.05 mM Cr(VI). Substrate analyses
revealed that although Cr(VI) was reduced within the first 5 h, growth was not observed for an additional 20 h. The growth
lag could be explained by a decline in cell viability; however, during this time small amounts of lactate were still utilized
without sulfate reduction or acetate formation. Approximately 40 h after Cr exposure (0.05 mM), sulfate reduction occurred
concurrently with the accumulation of acetate. Similar amounts of hydrogen were produced by Cr-exposed cells compared to control
cells, and lactate was not converted to glycogen during non-growth conditions. D. vulgaris cells treated with a reducing agent and then exposed to Cr(VI) still experienced a growth lag, but the addition of ascorbate
at the time of Cr(VI) addition prevented the lag period. In addition, cells grown on pyruvate displayed more tolerance to
Cr(VI) compared to lactate-grown cells. These results indicated that D. vulgaris utilized lactate during Cr(VI) exposure without the reduction of sulfate or production of acetate, and that ascorbate and
pyruvate could protect D. vulgaris cells from Cr(VI)/Cr(III) toxicity.
J.D. Wall and M.W. Fields are both affiliated to the Virtual Institute of Microbial Stress and Survival ().
M.E. Clark and S.B. Thieman contributed equally to this work. 相似文献
75.
The results of our present study indicate that 1 alpha, 25-dihydroxyvitamin D3[1 alpha, 25(OH)2D3] directly induces fusion of mouse alveolar macrophages without any participation of T-lymphocytes by a mechanism involving RNA and protein synthesis but not DNA synthesis. We have reported that 1 alpha, 25(OH)2D3 induces fusion of alveolar macrophages by a direct mechanism and by a spleen cell-mediated indirect mechanism [(1983) Proc. Natl. Acad. Sci. USA 80, 5583-5587]. Alveolar macrophages pretreated with or without anti-Thy 1.2 antibody and complement fused similarly when they were incubated with 1 alpha, 25(OH)2D3. The vitamin suppressed DNA synthesis, but it significantly enhanced RNA and protein synthesis. The 1 alpha, 25(OH)2D3-induced fusion was blocked by adding actinomycin D or cycloheximide, but not by hydroxyurea. 相似文献
76.
Yew-Hu Chien Wen-Chung Shiau 《Journal of experimental marine biology and ecology》2005,318(2):201-211
Natural carotenoids from astaxanthin containing alga Haematococcus pluvialis (H) and a non-astaxanthin carotenoid-containing alga Spirulina pacifica (S), and a synthetic astaxanthin Carophyll Pink (A) were supplemented in formulated diets at two concentrations, 50 (I) and 100 (II) mg kg−1, resulting in seven pigmented diets HI, SI, AI, HII, SII, AII, and HS (H-50 mg kg−1+S-50 mg kg−1). Formulated diet without carotenoid supplementation served as a control (C). The different diets were fed to juvenile kuruma prawn Marsupenaeus japonicus for 9 weeks. Dietary carotenoid effects on survival, growth, and pigmentation were compared by the treatment individually or collectively. A low dissolved oxygen stress test was conducted 2 weeks later and prawns' survival time and oxygen consumption rate were also compared among treatments. After 9 weeks' rearing, C-fed prawn had significantly lower survival rate than the pigmented diets-fed prawns. No difference in weight gain was found among all prawns. C-fed prawn had 66.4% less flesh astaxanthin (FA) and 75.5% less shell astaxanthin (SA) than the pigmented diets-fed prawns. I-fed (AI, HI, and SI) prawns had 31.1% less FA and 29.6% less SA than II-fed (AII, HII, SII, and HS) prawns. No significant differences were found in the comparisons by other categories. The use of these three sources of carotenoids for pigmentation in crustacean was discussed along carotenoid conversion, deposition, digestibility, and absorption. When subjected to low dissolved oxygen stress, C-fed prawn had higher oxygen consumption rate (OCR) and shorter survival time (ST) than the prawns fed the pigmented diets. No differences in OCR or ST were found in the comparisons by other categories. 相似文献
77.
The reaction of the bisguanidine copper(I) compounds [Cu(btmgp)I] and [Cu2(btmgp)2][PF6]2 with molecular oxygen afforded at low temperatures complexes containing the bis-μ-oxo dicopper(III) core, which is capable to hydroxylate one of the N-CH3-groups of the {bis(tetramethyl)guanidino}propane ligands. The formation of the novel ligand {bis(trimethylmethoxy)guanidino}propane (btmmO) is reported as it represents the first hydroxylation of a N-methyl group. The products of this reaction are novel alkoxo-bridged binuclear copper complexes, namely [Cu2(btmmO)2I]+ containing an iodide ion in a novel bridging situation, as well as [Cu2(btmmO)2]2+ which have been identified in their complex salts and [Cu2(btmmO)2][PF6]2 · 2MeCN, respectively. Concomitantly, the hydroxo-bridged binuclear copper compounds [Cu2(btmgp)2(μ-OH)2]I2 and [Cu2(btmgp)2(μ-OH)2][PF6]2 are formed as couple products. The formation of the bis-μ-oxodicopper(III) complexes was monitored by UV/Vis-spectroscopy, and the reaction products were characterised by X-ray diffraction, vibrational spectroscopy and elemental analysis. 相似文献
78.
The effect of excessive Cd on the growth and metal uptake by leafy vegetables Brassica chinensis L. (cv. Wuyueman) and Brassica pekinensis (Lour.) Rupr. (cv. Qingyan 87-114) were studied in hydroponic solution culture. The Cd concentration higher than 10 μM significantly
decreased the root elongation and leaf chlorophyll contents of both plant species. The shoots of B. pekinensis had significantly higher concentrations of total and water-soluble Cd than B. chinensis. The roots of both species accumulated more Cd than the shoots in all the Cd treatments. Most of the Cd in the roots was
found in the cell walls. The shoot/root ratio of Cd concentrations in B. pekinensis was always greater than that in B. chinensis. But, the concentration and proportion of Cd in the cell walls in B. chinensis were higher than that in B. pekinensis. Cadmium treatments also increased the concentrations of total non-protein thiols (NPT) in the shoots of the both species.
A significant correlation was found between the concentrations of soluble Cd and NPT in plant shoots. 相似文献
79.
Necrostatin-1 protects against glutamate-induced glutathione depletion and caspase-independent cell death in HT-22 cells 总被引:3,自引:0,他引:3
Xu X Chua CC Kong J Kostrzewa RM Kumaraguru U Hamdy RC Chua BH 《Journal of neurochemistry》2007,103(5):2004-2014
Glutamate, a major excitatory neurotransmitter in the CNS, plays a critical role in neurological disorders such as stroke and Parkinson's disease. Recent studies have suggested that glutamate excess can result in a form of cell death called glutamate-induced oxytosis. In this study, we explore the protective effects of necrostatin-1 (Nec-1), an inhibitor of necroptosis, on glutamate-induced oxytosis. We show that Nec-1 inhibits glutamate-induced oxytosis in HT-22 cells through a mechanism that involves an increase in cellular glutathione (GSH) levels as well as a reduction in reactive oxygen species production. However, Nec-1 had no protective effect on free radical-induced cell death caused by hydrogen peroxide or menadione, which suggests that Nec-1 has no antioxidant effects. Interestingly, the protective effect of Nec-1 was still observed when cellular GSH was depleted by buthionine sulfoximine, a specific and irreversible inhibitor of glutamylcysteine synthetase. Our study further demonstrates that Nec-1 significantly blocks the nuclear translocation of apoptosis-inducing factor (a marker of caspase-independent programmed cell death ) and inhibits the integration of Bcl-2/adenovirus E1B 19 kDa-interacting protein 3 (a pro-death member of the Bcl-2 family) into the mitochondrial membrane. Taken together, these results demonstrate for the first time that Nec-1 prevents glutamate-induced oxytosis in HT-22 cells through GSH related as well as apoptosis-inducing factor and Bcl-2/adenovirus E1B 19 kDa-interacting protein 3-related pathways. 相似文献
80.