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1.
The influence of ionic strength and composition on the binding and inhibition of human leukocyte elastase by glycosaminoglycans with variable degree and position of sulfation was investigated. The kinetic mechanism of inhibition had a hyperbolic, mixed-type character with a competitive component that was promoted by low ionic strength, reduced by phosphate ions, and which also depended on the substrate and glycosaminoglycan structure. Enzyme binding was a cooperative phenomenon that varied with ionic strength and composition. The inhibition patterns correlated with the cationic character of elastase and with the distribution of arginines on its molecular surface, most notably with residues located in the vicinity of the substrate binding region. The order of affinity for elastase binding was chondroitin 4-sulfate < chondroitin 6-sulfate < dermatan sulfate, iduronate-containing derivatives being superior with respect to the glucuronate-containing counterparts. Additional sulfation at both the 4- and 6- positions or at the N- and 4-positions of the N-acetylgalactosamine moiety decidedly improved the inhibitory efficiency. The results highlight a fundamental physiological role of enzyme-glycosaminoglycan interactions. In the azurophil granule of the human polymorphonuclear neutrophil, elastase and other enzymes are bound to a matrix of chondroitin 4-sulfate because this is the only glycosaminoglycan that simultaneously offers good binding for enzyme compartmentalization together with prompt release from the bound state at the onset of phagocytosis.  相似文献   
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The phenotypic characteristics of three Serpulina pilosicoli strains isolated from humans with diarrhoea (WesB, Kar, Hrm7) and two porcine S. pilosicoli strains isolated from pigs with intestinal spirochaetosis (1648, 3295), were compared with the type strain of the species P43/6/78T (T = type strain) and other intestinal spirochaetes within the genus Serpulina. All S. pilosicoli strains had a characteristic ultrastructural appearance, displayed similar growth rates, hydrolysed hippurate, lacked beta-glucosidase activity, utilised D-ribose as a growth substrate, and had similar sensitivities to rifampicin and spiramycin. The only consistent phenotypic characteristic that differentiated human strains from porcine strains of S. pilosicoli was that the human strains all utilised the pentose sugar D-xylose. These distinguishing phenotypic traits appear useful for identifying S. pilosicoli.  相似文献   
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The application of micro- and nanofabrication processes to the development of hybrid synthetic and biological systems may enable the production of new devices such as controllable transporters, gears, levers, micropumps, or microgenerators powered by biological molecular motors. However, engineering these hybrid devices requires fabrication processes that are compatible with biological materials such as kinesin motor proteins and microtubules. In this paper, the effects of micro- and nanofabrication processing chemicals and resists on the functionality of casein, kinesin, and microtubule proteins are systematically examined to address the important missing link of the biocompatibility of micro- and nanofabrication processes needed to realize hybrid system fabrication. It is found that both casein, which is used to prevent motor denaturation on surfaces, and kinesin motors are surprisingly tolerant of most of the processing chemicals examined. Microtubules, however, are much more sensitive. Exposure to the processing chemicals leads to depolymerization, which is partially attributed to the pH of the solutions examined. When the chemicals were diluted in aqueous buffers, a subset of them no longer depolymerized microtubules and in their diluted forms still worked as resist removers. This approach broadens the application of micro- and nanofabrication processes to hybrid synthetic and biological system fabrication.  相似文献   
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The L5178Y/tk+/- (-)3.7.2C mouse lymphoma assay (MLA) which detects mutations affecting the heterozygous thymidine kinase (tk) locus is capable of responding to chemicals acting as clastogens as well as point mutagens. Improvements in the assay to enhance detection of this spectrum of genetic events are summarized, and criteria for evaluating the data are defined. Using these criteria, the Phase III Work Group reviewed and evaluated literature containing MLA results published from 1976 through 1993. The data base included 602 chemicals of which 343 were evaluated as positive, 44 negative, 18 equivocal, 54 apparently inappropriate for evaluation in this test system with the published protocols, and 142 that were inadequately tested, and thus a definitive call could not be made. The overall performance of the assay is summarized by chemical class, and the outcome of testing 260 chemicals in the MLA is compared with Gene-Tox and National Toxicology Program evaluations of rodent carcinogenesis bioassay results for the same chemicals. Based on the Work Group's evaluation of published MLA data for chemicals that were considered adequately tested, it is concluded that for most chemicals the L5178Y/tk+/- mouse lymphoma assay is eminently well suited for genotoxicity testing and for predicting the potential for carcinogenicity.  相似文献   
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The suppressive effect of the halogenated inhalation anesthesia on cortical somatosensory evoked potentials (cSSEPs) has been well documented. Less studied and appreciated is the effect of nitrous oxide often with a narcotic as an alternative to a potent agent for spinal cord monitoring. This study sought to define more clearly the influence of nitrous oxide on cSSEPs elicited to posterior tibial nerve stimulation. A secondary purpose was to demonstrate the advantage of a total intravenous propofol anesthesia in facilitating uncompromised large-amplitude cSSEPs. Fifty adult patients undergoing anterior cervical discectomy served as the study sample. Brainstem and cortical posterior tibial nerve SSEPs were recorded under two independent anesthesia conditions, namely, nitrous oxide and propofol. Results demonstrated a significant amplitude reduction and latency prolongation with the nitrous oxide versus propofol protocol. cSSEP amplitude with propofol was, on the average, approximately two times larger than that with nitrous oxide. Based on these findings, the use of nitrous-oxide anesthesia is not recommended when limited to monitoring cSSEPs that are already amplitude compromised secondary to existing spinal cord disease.  相似文献   
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We have employed polyclonal antibodies to a peptide sequence of bovine steroidogenic acute regulatory (StAR) protein and human placental 3beta-hydroxysteroid dehydrogenase (3beta-HSD) to determine the localisation and distribution of these proteins in rat and bovine adrenal glands. Immunohistochemical staining demonstrated the presence of StAR protein in the zona glomerulosa (ZG), zona fasciculata (ZF), zona reticularis (ZR) and in the medulla of both species. For 3beta-HSD, immunostaining was observed in the ZG, ZF and ZR of the rat adrenal and was absent in the medulla. Immunoblotting experiments showed intense bands for StAR protein (30 kDa, 37 kDa) in the mitochondria of bovine ZG, ZF and medulla and a less intense band (30 kDa) in the microsomes. In rat ZG and ZF/R mitochondria only the 30 kDa protein was present. For 3beta-HSD, an intense band (42 kDa) was found in microsomes and mitochondria of rat and bovine ZG and ZFR. A very faint signal for 3beta-HSD was seen in adrenal medulla. In conclusion, StAR (or a closely related) protein is present throughout the adrenal gland in rat and bovine species in contrast to 3beta-HSD which is confined to the steroidogenic zones. The possible function of StAR protein in the adrenal medulla merits investigation.  相似文献   
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