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1.
Antibiotic treatment options for Burkholderia cepacia infection are limited because of high intrinsic resistance. The problem is complicated by development of cross-resistance between antibiotics of different classes. We isolated antibiotic-resistant mutants by stepwise exposure to chloramphenicol (Chlor) and to trimethoprim/sulphamethoxazole (T/S) for four B. cepacia strains: ATCC13945, Per (clinical isolate), Cas and D4 (environmental isolates). Chlor(r) mutants did not produce chloramphenicol acetyl-transferase. Cross-resistance, defined as greater than four-fold increase in MIC by microtitre dilution method, was consistently seen in both types of mutants. For chloramphenicol-resistant (Chlor[r]) and trimethoprim/sulphamethoxazole-resistant (Tr/Sr) mutants of B. cepacia ATCC13945 and Cas, no MIC change was seen for piperacillin, ceftazidime, rifampicin, gentamicin, tobramycin, polymyxin B or azithromycin. B. cepacia-Per and -D4 mutants showed cross-resistance to ceftazidime and to piperacillin. Comparison of outer membrane protein (OMP) profiles of B. cepacia and their mutants by SDS-PAGE revealed Tr/Sr) mutants to be deficient in a major OMP (molecular weight 39-47 kDa). Tr/Sr mutants also expressed additional OMPs not found in wild type strains at 75-77 kDa for B. cepacia-ATCC13945 and -Cas, and 20-21 kDa in B. cepacia-D4 and -Per. No OMP changes occurred in Chlor(r) mutants. Lipopolysaccharide (LPS) profiles of each type of mutant showed new high and low molecular weight LPS bands. Cross-resistance seems to be mediated by alterations in porin and LPS for Tr/Sr mutants, but only by LPS in Chlor(r) mutants. 相似文献
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The crystal structure of Escherichia coli ornithine transcarbamoylase (OTCase, EC 2.1.3.3) complexed with the bisubstrate analog N-(phosphonacetyl)-L-ornithine (PALO) has been determined at 2.8-A resolution. This research on the structure of a transcarbamoylase catalytic trimer with a substrate analog bound provides new insights into the linkages between substrate binding, protein-protein interactions, and conformational change. The structure was solved by molecular replacement with the Pseudomonas aeruginosa catabolic OTCase catalytic trimer (Villeret, V., Tricot, C., Stalon, V. & Dideberg, O. (1995) Proc. Natl. Acad. Sci. USA 92, 10762-10766; Protein Data Bank reference pdb 1otc) as the model and refined to a crystallographic R value of 21.3%. Each polypeptide chain folds into two domains, a carbamoyl phosphate binding domain and an L-ornithine binding domain. The bound inhibitor interacts with the side chains and/or backbone atoms of Lys-53, Ser-55, Thr-56, Arg-57, Thr-58, Arg-106, His-133, Asn-167, Asp-231, Met-236, Leu-274, Arg-319 as well as Gln-82 and Lys-86 from an adjacent chain. Comparison with the unligated P. aeruginosa catabolic OTCase structure indicates that binding of the substrate analog results in closure of the two domains of each chain. As in E. coli aspartate transcarbamoylase, the 240s loop undergoes the largest conformational change upon substrate binding. The clinical implications for human OTCase deficiency are discussed. 相似文献
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NM Bida 《Canadian Metallurgical Quarterly》1996,86(12):1555-1556
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DM Foreman S Jones NM McKechnie G Williams ME Boulton 《Canadian Metallurgical Quarterly》1996,28(5):296-302
We describe a simple method for combining in situ hybridisation and immunohistochemistry on the same retinal section. The technique was developed using a radiolabelled cDNA probe for opsin and an antibody (ROS1F4) against rhodopsin. This method retains the antigenic sites if immunocytochemistry is performed prior to in situ hybridisation. Opsin mRNA was found in the photoreceptor inner segment with rhodopsin immunolocalised to the photoreceptor outer segments. The technique should be applicable to numerous situations including analysis of the sequence of events in the expression and synthesis of the various opsins during retinal development and degeneration. 相似文献
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PR Bennett R Warwick J Vaughan H Chana A Lubenko NM Fisk 《Canadian Metallurgical Quarterly》1994,101(3):246-249
OBJECTIVE: To assess the effectiveness of a newly developed individualised birthweight ratio (IBR), which corrects for physiological birthweight determinants, in identifying infants at risk from the complications of macrosomia. DESIGN: Prospective observational study. SETTING: Obstetric unit, Nottingham City Hospital. SUBJECTS: 2835 women delivered between December 1991 and July 1992 and the infants of 624 of these, selected by virtue of their birthweight for gestation and IBR centile positions. MAIN OUTCOME MEASURES: Skinfold thickness and ponderal index measurements, operative delivery, shoulder dystocia, fetal trauma, impaired glucose tolerance. RESULTS: Using an IBR above the 90th centile as a cut off results in 2.4% of infants being reclassified as normally grown and 3.1% are reclassified as large. The IBR does not result in the identification of any more infants with abnormal ponderal indices or skinfold thicknesses than birthweight for gestation. It does, however, identify more of the infants at risk of operative delivery, shoulder dystocia, fetal trauma and impaired glucose tolerance. CONCLUSION: The IBR significantly improves upon birthweight for gestation in identifying infants who suffer from the complications of relative macrosomia. 相似文献
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J Desaki M Kinutani K Tan-Takeuchi NM Le Douarin 《Canadian Metallurgical Quarterly》1996,45(5):448-452
The migration of leucocytes through the walls of venules was examined in detail in the posterior latissimus dorsi muscle of quail-chick spinal cord chimeras, in which flaccid paralysis of the wings was observed. Examinations were made from the 70th to 80th day after hatching. Muscle fibers were degenerated and intramuscular nerve bundles destroyed. Massive leucocytes (almost lymphocytes) were found around the venules, depending on the passage of wandering leucocytes through the endothelium. Lymphocytes penetrated and were encased in the cytoplasm of the venular endothelial cell, and did not pass through the interendothelial junction. These findings suggest that, in the venules of the atrophied chimeric muscle, wandering leucocytes in the blood may pass through the endothelial cell body and migrate into the inflamed extravascular space. 相似文献