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91.
EP Guthrie T Quinton-Jager LS Moran BE Slatko RB Kucera JS Benner GG Wilson JE Brooks 《Canadian Metallurgical Quarterly》1996,180(1-2):107-112
SphI, a type II restriction-modification (R-M) system from the bacterium Streptomyces phaeochromogenes, recognizes the sequence 5'-GCATGC. The SphI methyltransferase (MTase)-encoding gene, sphIM, was cloned into Escherichia coli using MTase selection to isolate the clone. However, none of these clones contained the restriction endonuclease (ENase) gene. Repeated attempts to clone the complete ENase gene along with sphIM in one step failed, presumably due to expression of SphI ENase gene, sphIR, in the presence of inadequate expression of sphIM. The complete sphIR was finally cloned using a two-step process. PCR was used to isolate the 3' end of sphIR from a library. The intact sphIR, reconstructed under control of an inducible promoter, was introduced into an E. coli strain containing a plasmid with the NlaIII MTase-encoding gene (nlaIIIM). The nucleotide sequence of the SphI system was determined, analyzed and compared to previously sequenced R-M systems. The sequence was also examined for features which would help explain why sphIR unlike other actinomycete ENase genes seemed to be expressed in E. coli. 相似文献
92.
LS Wafelman BH Pollock J Kreutzer DS Richards AA Hutchison 《Canadian Metallurgical Quarterly》1999,75(2):73-81
Prognostic factors for survival of 62 fetuses and neonates with nonimmune hydrops fetalis (NIHF) were studied retrospectively. Twenty-eight infants survived >/=28 days which is 45% for all fetuses and newborns diagnosed with NIHF and 61% for liveborns with unresolved NIHF. Univariate analysis identified that mortality was associated with the presence of >/=2 serous cavity effusions and a need for chest compressions at birth. Multivariate logistic regression analysis confirmed that the presence of >/=2 serous cavity effusions was significantly associated with mortality from NIHF <28 days after birth [OR = 48.2 (CI 3.6, 662.9) (p < 0.004)]. We conclude that, compared to published cases from the 1970s and early 1980s, survival of liveborns with NIHF seems improved. The decrease in stillbirths is more notable. The severity of hydrops at birth is the key determinant for survival. 相似文献
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95.
Polygalacturonase-inhibiting proteins (PGIPs) with different specificities are expressed in Phaseolus vulgaris 总被引:2,自引:0,他引:2
A Desiderio B Aracri F Leckie B Mattei G Salvi H Tigelaar JS Van Roekel DC Baulcombe LS Melchers G De Lorenzo F Cervone 《Canadian Metallurgical Quarterly》1997,10(7):852-860
The pgip-1 gene of Phaseolus vulgaris, encoding a polygalacturonase-inhibiting protein (PGIP), PGIP-1 (P. Toubart, A. Desiderio, G. Salvi, F. Cervone, L. Daroda, G. De Lorenzo, C. Bergmann, A. G. Darvill, and P. Albersheim, Plant J. 2:367-373, 1992), was expressed under control of the cauliflower mosaic virus 35S promoter in tomato plants via Agrobacterium tumefaciens-mediated transformation. Transgenic tomato plants with different expression levels of PGIP-1 were used in infection experiments with the pathogenic fungi Fusarium oxysporum f. sp. lycopersici, Botrytis cinerea, and Alternaria solani. No evident enhanced resistance, compared with the resistance of untransformed plants, was observed. The pgip-1 gene was also transiently expressed in Nicotiana benthamiana with potato virus X (PVX) as a vector. PGIP-1 purified from transgenic tomatoes and PGIP-1 in crude protein extracts of PVX-infected N. benthamiana plants were tested with several fungal polygalacturonases (PGs). PGIP-1 from both plant sources exhibited a specificity different from that of PGIP purified from P. vulgaris (bulk bean PGIP). Notably, PGIP-1 was unable to interact with a homogeneous PG from Fusarium moniliforme, as determined by surface plasmon resonance analysis, while the bulk bean PGIP interacted with and inhibited this enzyme. Moreover, PGIP-1 expressed in tomato and N. benthamiana had only a limited capacity to inhibit crude PG preparations from F. oxysporum f. sp. lycopersici, B. cinerea, and A. solani. Differential affinity chromatography was used to separate PGIP proteins present in P. vulgaris extracts. A PGIP-A with specificity similar to that of PGIP-1 was separated from a PGIP-B able to interact with both Aspergillus niger and F. moniliforme PGs. Our data show that PGIPs with different specificities are expressed in P. vulgaris and that the high-level expression of one member (pgip-1) of the PGIP gene family in transgenic plants is not sufficient to confer general, enhanced resistance to fungi. 相似文献
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LS Sheldon JT Keever JM Roberds JB Beach JN Morgan 《Canadian Metallurgical Quarterly》1997,7(1):37-59
Dietary uptake may be a significant pathway of exposure to contaminants. As such, dietary exposure assessments should be considered an important part of the total exposure assessment process. The objective of this work was to develop reliable methods that are applicable to a wide range of base/neutral and carbamate-type pesticides in duplicate diet samples collected as part of dietary exposure assessment studies. The resulting method needed to be sensitive to concentrations below 1 ng/g, accurate and precise, and as simple and cost effective as possible. As a first step, information was gathered on current methods for measuring pesticides in foods. Although the literature methods could serve as a starting point, few had been applied to duplicate diet samples and detection limits were generally high (10 to 100 ng/g). Experimental work was performed to evaluate individual extraction, cleanup, and analysis procedures; link the most promising procedures into analysis methods; and generate performance data on the final method. The final method used Soxhlet extraction with solvent partitioning and gel permeation chromatography cleanup. Gas chromatography/mass spectrometry was used for the analysis of base/neutral pesticides. High performance liquid chromatography analysis was used for the analysis of carbamate pesticides. Results of performance testing showed good accuracy (recovery > 70%), precision (% RSD < 25%), and sensitivity (method detection limits < 1.0 ng/g) for most pesticides targeted for study. 相似文献
98.
LS Lohmander 《Canadian Metallurgical Quarterly》1997,11(4):711-726
Biochemical markers for osteoarthritis (OA) may serve different purposes. Since markers reflect ongoing dynamic metabolic processes in the joint tissues (cartilage, synovium, bone, etc.), they are most likely to be useful to predict prognosis and response to treatment, to monitor response to treatment, and for disease staging. Markers are currently being used at the research level for these purposes. The goal of using these markers to assess the disease process in the OA clinical trial setting or in the clinical routine has, however, not yet been reached. 相似文献
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100.
J Wang J Kwon X Ding JA Fairley DT Woodley LS Chan 《Canadian Metallurgical Quarterly》1997,150(6):1901-1907
Intraepidermal neutrophilic IgA dermatosis, a rare skin disease entity manifested with blisters and pustules clinically and lower epidermal blister, acantholysis, and neutrophilic infiltration pathologically, was first reported in 1985. Although the disease is characterized by IgA autoantibodies targeting the epithelial cell surface component, the target antigen has not been determined. We investigated a patient with this disease by histopathology, direct and indirect immunofluorescence, immunoblotting, and immunoadsorption studies. The pustular lesion was characterized by blister at the lower epidermis, acantholysis, and neutrophilic infiltration. Nonsecretory IgA1 subclass autoantibodies targeting the lower epithelial cell surfaces were detected in the patient's skin and serum. The patient's IgA autoantibodies labeled a recombinant desmosomal protein desmoglein 3 on immunoblotting and the immunolabeling of epithelial cell surfaces was eliminated by preadsorption with desmoglein 3. Thus, desmoglein 3 is identified as a target antigen in intraepidermal neutrophilic IgA dermatosis. The ability of IgA1 autoantibodies to bind neutrophils may be responsible for the prominent neutrophilic infiltration observed histopathologically and for the pustular lesions observed clinically. 相似文献