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991.
Matthew R Litman Norman HL Chiu James Wang 《Journal of chemical technology and biotechnology (Oxford, Oxfordshire : 1986)》2006,81(3):318-321
The objective of this research project is to develop a rapid molecular method for monitoring nitrification in a wastewater reactor. In the developed method, a monoclonal antibody (Hyb 153‐3) was used because it can specifically recognize non‐denatured enzymes responsible for nitrite oxidation in Nitrobacter and Nitrospira. The recognition of these enzymes under non‐denatured conditions can significantly simplify the procedures of future immunoassays for environmental samples collected from various natural and engineered systems. This paper presents the ability of the selected Hyb 153‐3 antibody to recognize the non‐denatured form of the nitrite‐oxidizing enzyme of Nitrospira in an aqueous phase as well as when the enzyme has been immobilized on a solid surface. Copyright © 2005 Society of Chemical Industry 相似文献
992.
993.
Promiscuous liberation of MHC-class I-binding peptides from the C termini of membrane and soluble proteins in the secretory pathway 总被引:1,自引:0,他引:1
HL Snyder I Bacík JW Yewdell TW Behrens JR Bennink 《Canadian Metallurgical Quarterly》1998,28(4):1339-1346
TAP can efficiently transport peptides up to twice as long as those bound to MHC class I molecules, suggesting a role for endoplasmic reticulum (ER) proteases in the trimming of TAP-transported peptides. To better define ER processing of antigenic peptides, we examined the capacity of TAP-deficient cells to present determinants derived from ER-targeted proteins encoded by recombinant vaccinia viruses. TAP-deficient cells failed to present antigenic peptides from internal locations in secreted proteins to MHC class I-restricted T lymphocytes. The same peptides were liberated from the C termini of a secreted protein and the lumenal domains of two membrane proteins delivered to the ER via different routes. These findings suggest that proteases in the secretory compartment can liberate C-terminal antigenic peptides from virtually any context. We propose that this activity often participates in the removal of N-terminal extensions from TAP-transported peptides, thereby creating optimally sized products for MHC class I binding. We further demonstrate that ER trimming of C termini can occur if we express an appropriate carboxypeptidase in the secretory pathway. The absence of such trimming under normal circumstances suggests that carboxypeptidase activity is generally deficient in the ER, consistent with the concordance between the specificity of TAP and MHC class I molecules for the same types of C-terminal residues. 相似文献
994.
Although recent fMRI and single unit recording studies have shown that attention modulates neural activity in motion sensitive areas of extrastriate cortex, these approaches cannot reveal qualitative or quantitative effects of attention on perception of motion. To investigate this, we asked observers to select one of two orthogonal directions in a brief, transparent dot display (prime) and then measured their sensitivity to global directional motion in a second uni-directional dot display (probe) presented a short time later. When probe direction matched the attended prime direction, sensitivity was degraded. But, when probe direction matched the ignored prime direction, sensitivity was enhanced, even though both components were of equal physical strength. Sensitivity was unchanged for directions opposite to either previously seen direction. Neither sensory adaptation nor opponent direction mechanisms can account for these data. Rather, processes initiated by visual selection must underlie these dramatic changes in motion sensitivity. 相似文献
995.
HL Galan MJ Hussey M Chung JK Chyu JC Hobbins FC Battaglia 《Canadian Metallurgical Quarterly》1998,178(3):451-456
Crush syndrome is the severe systemic manifestation of prolonged muscle compression and compartment syndrome. Careful patient assessment, early diagnosis, and aggressive treatment are vital to prevent multiorgan failure and death. Medical management of systemic complications, along with operative procedures of fasciotomy and debridement, are indicated with accompanying compartment syndrome. Debridement of necrotic and nonviable tissue is necessary; significant risks of infection and hemorrhage remain until the wounds can be subsequently closed or covered with skin graft. Crush syndrome and muscle necrosis in a closed injury without compartment syndrome may be followed clinically until healing or demarcation of a gangrenous part occurs, providing the patient's general medical condition, including renal function, can be maintained. Fasciotomy and hyperbaric oxygen will not reverse necrosis of muscle in the absence of compartment syndrome and therefore do not affect outcome of the extremity. Overall, prognosis is improved by early diagnosis and treatment, but outcome of the crushed extremity is poor and Volkmann's contracture often results. 相似文献
996.
Mutations in a number of cardiac sarcomeric protein genes cause hypertrophic cardiomyopathy (HCM). Previous findings indicate that HCM-causing mutations associated with a truncated cardiac troponin T (TnT) and missense mutations in the beta-myosin heavy chain share abnormalities in common, acting as dominant negative alleles that impair contractile performance. In contrast, Lin et al. [Lin, D., Bobkova, A., Homsher, E. & Tobacman, L. S. (1996) J. Clin. Invest. 97, 2842-2848] characterized a TnT point mutation (Ile79Asn) and concluded that it might lead to hypercontractility and, thus, potentially a different mechanism for HCM pathogenesis. In this study, three HCM-causing cardiac TnT mutations (Ile79Asn, Arg92Gln, and DeltaGlu160) were studied in a myotube expression system. Functional studies of wild-type and mutant transfected myotubes revealed that all three mutants decreased the calcium sensitivity of force production and that the two missense mutations (Ile79Asn and Arg92Gln) increased the unloaded shortening velocity nearly 2-fold. The data demonstrate that TnT can alter the rate of myosin cross-bridge detachment, and thus the troponin complex plays a greater role in modulating muscle contractile performance than was recognized previously. Furthermore, these data suggest that these TnT mutations may cause disease via an increased energetic load on the heart. This would represent a second paradigm for HCM pathogenesis. 相似文献
997.
998.
CS Bonder HL Dickensheets JJ Finlay-Jones RP Donnelly PH Hart 《Canadian Metallurgical Quarterly》1998,160(8):4048-4056
IL-4 has potent anti-inflammatory properties on monocytes and suppresses both IL-1beta and TNF-alpha production. Well-characterized components of the IL-4 receptor on monocytes include the 140-kDa alpha-chain and the IL-2R gamma-chain, gammac, which normally dimerize 1:1 for signaling from the receptor. However, mRNA levels for gammac were very low in 7-day-cultured monocytes. As mRNA levels for gammac declined with culture, so too did the ability of IL-4 to down-regulate LPS-induced TNF-alpha production. In contrast, IL-4 consistently down-regulated IL-1beta production by cultured monocytes. Immunoprecipitation and Western blot analyses demonstrated that 7-day-cultured monocytes do not express the functionally active 64-kDa gammac protein. This was associated with decreased STAT6 activation by IL-4. Studies with Abs to gammac and an IL-4 mutant that is unable to bind to gammac showed that IL-4 can suppress IL-1beta but not TNF-alpha production by LPS-stimulated monocytes in the presence of little or no functioning gammac. IL-4 also suppressed IL-1beta but not TNF-alpha production by Mono Mac 6 cells, which express minimal levels of gammac. For gammac-expressing LPS/PMA-activated U937 cells, IL-4 decreased both TNF-alpha and IL-1beta production. These results suggest that functional gammac is not present on in vitro-derived macrophages, and that while some anti-inflammatory responses to IL-4 are lost with this down-regulation of functional gammac, others are retained. We conclude that different functional responses to IL-4 by human monocytes and macrophages are regulated by different IL-4 receptor configurations. 相似文献
999.
HL Hui JS Kavanaugh ML Doyle A Wierzba PH Rogers A Arnone JM Holt GK Ackers RW Noble 《Canadian Metallurgical Quarterly》1999,38(3):1040-1049
Human hemoglobin produced in the Escherichia coli coexpression system of Hernan et al. [(1992) Biochemistry 31, 8619-8628] has been transformed into a functionally homogeneous protein whose properties closely approximate those of normal hemoglobin A. Both of the alpha and beta chains of this hemoglobin contain a valine-methionine substitution at position 1 in order to accommodate the difference in specificity of the protein-processing enzymes of procaryotes. Despite extensive purification, functional homogeneity of the E. coli expressed hemoglobin was achieved only by the complete disassembly of the hemoglobin into its component alpha and beta globins and their reassembly in the presence of hemin. The kinetics of CO combination and the thermodynamics of O2 binding and cooperativity of the reassembled alphaV1M-betaV1M hemoglobin closely approximate those of HbA. The alpha globin obtained from the E. coli expressed hemoglobin was also combined with normal human beta chains and hemin to form the alphaV1M variant. The alpha+M variant of HbA, in which the normal N-terminal valine of the alpha chains is preceded by a methionine residue, was prepared by the same procedure. The kinetics of the reactions of CO with the alphaV1M and alpha+M variants are similar to those for HbA. The equilibria of oxygen binding to alphaV1M and HbA are similar whereas alpha+M exhibits a significantly higher oxygen affinity. The three-dimensional structures of alphaV1M and alpha+M offer an explanation for the latter affinity difference. Although the structures of alphaV1M and HbA, which have been determined by X-ray crystallography, are virtually indistinguishable except at the N-terminal residues, that of alpha+M indicates the displacement of a solvent molecule, possibly a chloride ion, from arginine 141alpha. Such an alteration in an anion binding site could result in increased oxygen affinity. 相似文献
1000.
HL Sham C Zhao KD Stewart DA Betebenner S Lin CH Park XP Kong W Rosenbrook T Herrin D Madigan S Vasavanonda N Lyons A Molla A Saldivar KC Marsh E McDonald NE Wideburg JF Denissen T Robins DJ Kempf JJ Plattner DW Norbeck 《Canadian Metallurgical Quarterly》1996,39(2):392-397
The design, synthesis, and molecular modeling studies of a novel series of azacyclic ureas, which are inhibitors of human immunodeficiency virus type 1 (HIV-1) protease that incorporate different ligands for the S1', S2, and S2' substrate-binding sites of HIV-1 protease are described. The synthesis of this series is highly flexible in the sense that the P1', P2, and P2' residues of the inhibitors can be changed independently. Molecular modeling studies on the phenyl ring of the P2 and P2' ligand suggested incorporation of hydrogen-bonding donor/acceptor groups at the 3' and 4-positions of the phenyl ring should increase binding potency. This led to the discovery of compound 7f (A-98881), which possesses high potency in the HIV-1 protease inhibition assay and the in vitro MT-4 cell culture assay (Ki = approximately 5 pM and EC50 = 0.002 microM). This compares well with the symmetrical cyclic urea 1 pioneered at DuPont Merck. 相似文献