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131.
A rabbit model of invasive aspergillosis has been used to investigate the pathogenesis of Aspergillus infection in the immunosuppressed host. The animals received hydrocortisone daily and a single dose of cyclophosphamide 2 days prior to intratracheal instillation of conidia from Aspergillus fumigatus. Bronchoalveolar lavage (BAL) was performed in 3 infected and 2 control saline treated animals sacrificed on days 1, 2, 4, 7 and 10 following inoculation. Infective load within the lung was quantified using an assay for chitin which is an important component of fungal cell walls (in particular the hyphal cell wall) and is not present in vertebrate tissue. The total BAL white cell count did not discriminate between infected and saline treated animals and Aspergillus was cultured from one lavage specimen only. Infected animals developed a marked neutrophil alveolitis by day 2 in contrast to a near total absence of neutrophils in the lavages of the control animals. Phagocytosis of conidia by alveolar macrophages was prominent but did not prevent progressive infection as confirmed by measurement of lung chitin. This pattern of cellular response within the alveolar airspace reflects the complex nature of the response to Aspergillus infection in the immunosuppressed host. 相似文献
132.
Rolf Mentlein Cornelia Buchholz Prof. Dr. Brigitte Krisch 《Cell and tissue research》1989,258(2):309-317
Summary The synthetic peptides somatostatin (SRIF) and growth hormone-releasing hormone (GRH) were coupled directly to colloidal gold of different particle sizes. Both conjugates were biologically active in displacing the corresponding radiolabeled hormones from high affinity binding sites in pituitary membranes. Release of growth hormone (GH) from cultured anterior pituitary cells was modulated by both conjugates alone or in combination. Ultrastructural studies were performed with cells incubated at 4° C (2 h) and 37° C (2 min-2 h) with one of the labeled peptides or their combination. Somatotropes were identified by immunostaining with anti-rGH followed by protein A-ferritin, thus obtaining a triple labeling. Both hormone conjugates were internalized in different vesicles in the beginning but accumulated during longer incubation times in the same compartment. The secretory vesicles and the nucleus were not labeled by any hormone conjugate. In contrast to SRIF-gold, the uptake of GRH-gold conjugate decreased with longer incubation times. This effect could be neutralized by simulatenous incubation of the somatotropes with both regulating hormones. Hence, whereas the binding and internalization of SRIF by somatotropes do not seem to be influenced by GRH, the corresponding processes for GRH are stimulated by the presence of SRIF. 相似文献
133.
Curtis W. Hoganson Demetrios F. Ghanotakis Gerald T. Babcock Charles F. Yocum 《Photosynthesis research》1989,22(3):285-293
Manganese in the oxygen-evolving complex is a physiological electron donor to Photosystem II. PS II depleted of manganese may oxidize exogenous reductants including benzidine and Mn2+. Using flash photolysis with electron spin resonance detection, we examined the room-temperature reaction kinetics of these reductants with Yz
+, the tyrosine radical formed in PS II membranes under illumination. Kinetics were measured with membranes that did or did not contain the 33 kDa extrinsic polypeptide of PS II, whose presence had no effect on the reaction kinetics with either reductant. The rate of Yz
+ reduction by benzidine was a linear function of benzidine concentration. The rate of Yz
+ reduction by Mn2+ at pH 6 increased linearly at low Mn2+ concentrations and reached a maximum at the Mn2+ concentrations equal to several times the reaction center concentration. The rate was inhibited by K+, Ca2+ and Mg2+. These data are described by a model in which negative charge on the membrane causes a local increase in the cation concentration. The rate of Yz
+ reduction at pH 7.5 was biphasic with a fast 400 s phase that suggests binding of Mn2+ near Yz
+ at a site that may be one of the native manganese binding sites.Abbreviations PS II
Photosystem II
- YD
tyrosine residue in Photosystem II that gives rise to the stable Signal II EPR spectrum
- Yz
tyrosine residue in Photosystem II that mediates electron transfer between the reaction center chlorophyll and the site of water oxidation
- ESR
electron spin resonance
- DPC
diphenylcarbazide
- DCIP
dichlorophenolindophenol 相似文献
134.
The temperature dependence of the partial reactions leading to turn-over of the UQH2:cyt c
2 oxidoreductase of Rhodobacter sphaeroides have been studied. The redox properties of the cytochrome components show a weak temperature dependence over the range 280–330 K, with coefficients of about 1 m V per degree; our results suggest that the other components show similar dependencies, so that no significant change in the gradient of standard free-energy between components occurs over this temperature range. The rates of the reactions of the high potential chain (the Rieske iron sulfur center, cytochromes c
1 and c
2, reaction center primary donor) show a weak temperature dependence, indicating an activation energy < 8 kJ per mole for electron transfer in this chain. The oxidation of ubiquinol at the Qz-site of the complex showed a strong temperature dependence, with an activation energy of about 32 kJ mole–1. The electron transfer from cytochrome b-566 to cytochrome b-561 was not rate determining at any temperature, and did not contribute to the energy barrier. The activation energy of 32 kJ mole–1 for quinol oxidation was the same for all states of the quinone pool (fully oxidized, partially reduced, or fully reduced before the flash). We suggest that the activation barrier is in the reaction by which ubiquinol at the catalytic site is oxidized to semiquinone. The most economical scheme for this reaction would have the semiquinone intermediate at the energy level indicated by the activation barrier. We discuss the plausibility of this simple model, and the values for rate constants, stability constant, the redox potentials of the intermediate couples, and the binding constant for the semiquinone, which are pertinent to the mechanism of the ubiquinol oxidizing site.Abbreviations (BChl)2
P870, primary donor of the photochemical reaction center
-
b/c
1 complex
ubiquinol: cytochrome c
2 oxidoreductase
- cyt b
H
cytochrome b-561 or higher potential cytochrome b
- cyt b
L
cytochrome b-566, or low potential cytochrome b
- cyt c
1, cyt c
2, cyt c
t
cytochromes c
1 and c
2, and total cytochrome c (cyt c
1 and cyt c
2)
- Fe.S
Rieske-type iron sulfur center, Q
- QH2
ubiquinone, ubiquinol
- Qz, QzH2, Qz
–
ubiquinone, ubiquinol, and semiquinone anion of ubiquinone, bound at quinol oxidizing site
- Qz-site
ubiquinol oxidizing site (also called Qo-(outside)
- Qo
(Oxidizing)
- QP
(Positive proton potential) site)
- Qc-site
uubiquinone reductase site (also called the Qi-(inside)
- QR
(Reducing), or
- QN
(Negative proton potential) site)
- UHDBT
5-(n-undecyl)-6-hydroxy-4,7-dioxobenzothiazol 相似文献
135.
Francisco Javier Caballero Isabel Igeño Jacobo Cárdenas Francisco Castillo 《Archives of microbiology》1989,152(5):508-511
The phototrophic bacterium Rhodobacter capsulatus E1F1 assimilates ammonia and other forms of reduced nitrogen either through the GS/GOGAT pathway or by the concerted action of l-alanine dehydrogenase and aminotransferases. These routes are light-independent and very responsive to the carbon and nitrogen sources used for cell growth. GS was most active in cells grown on nitrate or l-glutamate as nitrogen sources, whereas it was heavily adenylylated and siginificantly repressed by ammonium, glycine, l-alanine, l-aspartate, l-asparagine and l-glutamine, under which conditions specific aminotransferases were induced. GOGAT activity was kept at constitutive levels in cells grown on l-amino acids as nitrogen sources except on l-glutamine where it was significantly induced during the early phase of growth. In vitro, GOGAT activity was strongly inhibited by l-tyrosine and NADPH. In cells using l-asparagine or l-aspartate as nitrogen source, a concerted induction of l-aspartate aminotransferase and l-asparaginase was observed. Enzyme level enhancements in response to nitrogen source variation involved de novo protein synthesis and strongly correlated with the cell growth phase.Abbreviations ADH
l-alanine dehydrogenase
- AOAT
l-alanine:2-oxoglutarate aminotransferase
- Asnase
l-asparaginase
- GOAT
Glycine: oxaloacetate aminotransferase
- GOGAT
Glutamate synthase
- GOT
l-aspartate: 2-oxoglutarate aminotransferase
- GS
Glutamine synthetase
- HPLC
High-Pressure Liquid Chromatography
- MOPS
2-(N-morpholino)propanesulfonic acid
- MSX
l-methionine-d,l-sulfoximine 相似文献
136.
I. M. Birk R. Dierstein I. Kaiser U. Matern W. A. König R. Krebber J. Weckesser 《Archives of microbiology》1989,151(5):411-415
Toxic and nontoxic peptides were isolated from the cyanobacterium Microcystis aeruginosa PCC 7806 by a procedure including extraction of cells with water-saturated 1-butanol, chromatography of the extract on silica
gel plates and high performance liquid chromatography (HPLC) on Partisil-5. The toxin was shown to be only a minor constituent,
being negatively charged and thus separable by electrophoresis, within the HPLC-purified fraction. It contained erythro-β-methyl-D-Asp, D-Glu, D-Ala, L-Leu, and L-Arg known to be part of the Microcystis peptide-toxin with Mr 994. The major part of the HPLC-purified fraction was assigned, however, to a nontoxic peptide with a Mr of 956. Partial hydrolysis studies of the nontoxic peptide(s) revealed amino acid sequences composed of D-Glu, N-methyl-Phe, and 3,4-dehydro-Pro, aside from the common L-amino acids. Cyclic linkage in the nontoxic peptide(s) appears likely. 相似文献
137.
138.
Fucosidosis is an autosomal recessive lysosomal storage disease due to a deficiency of-L-fucosidase activity in tissues and body fluids. Exponentially growing lymphoid cell cultures from four fucosidosis patients had 2.7-fold to 15.6-fold less extracellular-L-fucosidase protein and 28.8-fold to 144.0-fold less intracellular-L-fucosidase protein with negligible catalytic activity, compared to the mean of 19 control cultures. The percentage of total-L-fucosidase protein released extracellularly by cultures from the four patients was 64 to 85%, compared to 35±9% for control cultures. Intracellular and extracellular enzyme forms in fucosidosis and control cell lines were glycoproteins containing polypeptide chains ofM
r=52,000. During a 1.5-hr pulse-label with35S-methionine,-L-fucosidase was synthesized by control cells and two fucosidosis cell lines as an intracellular form withM
r=58,000. During a subsequent 21-hr chase with unlabeled methionine, mutant enzyme was almost entirely processed to an extracellular form withM
r=62,000. In contrast, only 25–30% of control enzyme was processed to an extracellular form (M
r=62,000), with the remainder retained intracellularly (M
r=60,000). In the other two fucosidosis cell lines,-L-fucosidase was synthesized as an intracellular form withM
r=56,000 that was processed to an extracellular form withM
r=60,000. In summary, the fucosidosis mutation(s) affected the catalytic activity, quantity, and extracellular release of-L-fucosidase as expressed by lymphoid cells.This work was funded by NIH Grants DK 32161 to R. A. DiCioccio and GM 28428 to J. K. Darby. 相似文献
139.
The combined effects of chlorine and ammonia on litter breakdown in outdoor experimental streams 总被引:2,自引:0,他引:2
The response of Potamogeton crispus L. breakdown to controlled doses of different levels of chlorine and chlorine + ammonia was investigated over two years in
outdoor experimental streams. In 1985, downstream riffles of 2 streams were dosed (observed in-stream concentrations) at ca.
10 μg/L Total Residual Chlorine (TRC), one stream at 64 μg/L TRC and one stream at 230 μg/L TRC. Two control streams were
not dosed and the upstream riffles of each stream served as within stream controls. In 1986, the downstream riffle of one
stream was dosed at 70 μg/L TRC and a second stream was dosed at 200 μg/L TRC. Four streams were also dosed with 2.5 mg/L
NH3-N: one stream with no chlorine, one stream with ca. 10 μg/L TRC, one with 56 μg/L TRC, and one with 150 μg/L TRC. A seventh
stream was dosed for 2 h at 2000 μg/L TRC and 2.5 mg/L ammonia and then allowed to recover (recovery stream). Each year, litter
decomposition (degree day k values) was measured during two 35 day trials (Jun–Jul and Aug–Sep). In 1985, when streams were dosed with chlorine alone,
decomposition was significantly reduced with the high (230 μg/L TRC) chlorine dose. Downstream decomposition was 27% (Jun–Jul)
and 59% (Aug–Sep) of the upstream (control) rate. No other chlorine effects were found during this period. In Jun–Jul 1986,
there was significantly lower decomposition in the downstream dosed sites of the 200 μg/L TRC alone stream, the 146 μg/L TRC
+ ammonia stream and the recovery stream; downstream decay rates were (respectively) 56%, 42% and 64% of the upstream control
sites. No other up-down pairs were different in July 1986. In Aug–Sep, all three streams with chlorine + ammonia (6, 56 and
146 μg/L TRC + 2,5 mg/L ammonia) and the 70 μg/L TRC alone stream had significantly lower decomposition rates in the downstream
dosed sites. For these streams, downstream decay rates ranged from 46% (high chlorine + ammonia) to 73% (low chlorine + ammonia)
of the upstream control rates. No other up-down pairs were different during this trial. Up and downstream sites of the stream
dosed with 2.5 mg/L ammonia alone were nearly identical for both trials (< 3% difference). These results indicate that TRC
at less than 250 μg/L can significantly reduce litter decomposition and strongly suggest that addition of ammonia to chlorinated
water can increase the toxic effect of chlorine.
currently at the Department of Fisheries and Wildlife
currently at the Department of Fisheries and Wildlife 相似文献
140.
Richard T. Sawyer 《Mycopathologia》1990,109(2):99-109
The initial interaction of Candida albicans with pulmonary tissue of B6D2/F1 mice was investigated. The LD50 for mice challenged intravenously (IV) was approximately 3 × 105 yeasts, whereas the LD50 by the intratracheal (IT) route was in excess of 108 yeasts. Mice challenged IV died of progressive yeast growth in the kidneys. In contrast, mice challenged IT rapidly eliminated the entire inoculum by the first day after challenge. Resident pulmonary alveolar macrophages (PAM) killed upwards of 70% of C. albicans in an in vitro killing assay. At effector: target ratios favoring the effector cell population resident PAM were able to restrict the formation of yeast germ tubes to only 30% of the yeasts, whereas at equivalent ratios virtually all of the intracellular yeasts produced germ tubes. Evaluation of the ability of PAM, harvested from genetically different strains of inbred mice, to kill C. albicans in vitro showed that killing ability was a property of resident PAM from mice with the black 6 background. It was discovered that during the initial stages of infection in vivo, the expression of the F4/80 surface molecule was down regulated, and the expression of the Mac 1 surface molecule upregulated. There were no quantitative changes in expression of either Mac 2, Mac 3, Ly 5 or the 5C6 surface epitopes. Taken together, the data show that pulmonary tissue is quantitatively very resistant to C. albicans infection, because of the ability of resident PAM to rapidly phagocytize and kill yeasts. Killing of C. albicans by resident PAM may be a property of a subset of this mononuclear phagocyte population and was accompanied by alterations in the expression of surface molecules.Presented as part of the Everett S. Beneke Symposium in Mycology, May 27, 1988. 相似文献