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851.
Characterization of N-linked oligosaccharides attached to recombinant human antithrombin expressed in the yeast Pichia pastoris 总被引:2,自引:0,他引:2
We studied the structures of four N-linked oligosaccharide chains of the recombinant human antithrombin (rAT) expressed in the yeast Pichia pastoris. rAT was fully glycosylated at Asn 96 and Asn 155, whereas the glycosylation on Asn 135 and Asn 192 was partial. The glycosylation level on Asn 135 was only 12% and this reduction is assumed to be one of the reasons for a higher heparin-binding affinity of rAT than plasma-derived human antithrombin (pAT). In order to determine the sizes and electrostatic charges of the N-linked oligosaccharides, rAT was treated with PNGase F, and the reduced ends were labelled by pyridylamination followed by analysis using anion exchange and amide adsorption columns. The N-linked oligosaccharides were 78% neutral and 22% phosphomannosylated. The neutral oligosaccharides were thought to be Man(9-12)GlcNAc(2) as their major components. The phosphomannosylated oligosaccharides were then subjected to mild acid hydrolysis and/or digestion with alkaline phosphatase, and their charge shifts were analysed by the affinity to an anion exchange column. Among phosphomannosylated oligosaccharides, monophosphate diester type was predominant, whereas negatively charged diphosphate diester and monophosphate monoester types were minor components. The mannose residues at the non-reducing end(s) of Man(9-12)GlcNAc(2) were phosphomannosylated or phosphorylated and these are the major components. Because rAT is less negatively charged than pAT, which has disialyl biantennary N-glycans, it might be less repulsive to pentasaccharide-bearing anticoagulantly active heparan sulphate proteoglycan molecules exposed on the surface of the damaged vascular vessels. 相似文献
852.
Development of an automation system for single nucleotide polymorphisms genotyping using bio-strand, a new three-dimensional microarray 总被引:1,自引:0,他引:1
Tojo Y Asahina J Miyashita Y Takahashi M Matsumoto N Hasegawa S Yohda M Tajima H 《Journal of Bioscience and Bioengineering》2005,99(2):120-124
Previously, we developed a novel three-dimensional microarray system called Bio-Strand, which may be used in various applications including single nucleotide polymorphisms genotyping. In Bio-Strand, samples for detection are immobilized on a one-dimensional thread, which is wound around a cylinder-shaped core to form a three-dimensional thread-and-core structure. The thread-and-core structure is then inserted into a plastic pipette tip, where hybridization and detection are performed. In this study, we have developed an automation system, NIAGALA Bio-Station SDx, which enables automated hybridization and detection during the genotyping procedure using Bio-Strand. Using this system, we have performed the single nucleotide polymorphism (SNP) genotyping of CYP2C, one of the important human cytochrome P450 genes and the results were completely consistent with the genotyping results determined by the TaqMan method. 相似文献
853.
P.L. Nilantha Lakshman Yoichi Toyokawa Hirohide Toyama Toki Taira Masaaki Yasuda 《Food chemistry》2010
Genus Monascus is one of the most important microorganisms in the fermentation industry in Asia. However, only a little attention has been paid to the proteinases produced by this fungus and their role in the fermentation process. The main objective of this study was to purify and characterise acid proteinases produced by Monascus pilosus. Two acid proteinases (MpiAP1 and MpiAP2) were purified to homogeneity. Both purified enzymes, MpiAP1 and MpiAP2, were monomeric structures with molecular masses of around 43 and 58 kDa, respectively. The former was an acidic non-glycoprotein, whereas the latter was an acidic glycoprotein with 27% carbohydrate content. Although amino-terminal amino acid sequence analysis of both enzymes (MpiAP1 and MpiAP2) of 20 amino acid length showed over 90% similarity, their amino-terminal amino acids were different from each other. Both enzymes were optimally active at 55 °C and at pH 2.5–3.0 against casein or human haemoglobin. The T1/2 values of MpiAP1 and MpiAP2 were 65 and 70 °C, respectively. Both of the enzymes were completely inhibited by pepstatin A, and markedly by SDS. MoO3 also showed a partial inhibition of both enzymes. Milk casein and haemoglobin were good substrates for these enzymes. Eleven cleavages were detected using the oxidised insulin B-chain as a peptide for the proteolytic specificity test of MpiAP1, while seven cleavages were detected for MpiAP2. 相似文献
854.
Omori Y Sakikubo T Nakane M Fuchu H Miake K Kodama Y Sugiyama M Nishikawa Y 《Journal of food protection》2010,73(10):1803-1808
To manufacture raw ham in an efficient manner, we recently developed a new system in which presliced pork loin was used, and the processing time was reduced to 5% of the conventional method. This study aimed to examine whether this raw ham could be as safe as ham produced by the conventional method. Pork loin spiked with enterohemorrhagic Escherichia coli serotype O157:H7, Listeria monocytogenes serotype 1/2c, Salmonella enterica serovar Enteritidis, and Staphylococcus aureus were processed using either the new or conventional method. The fate of the foodborne pathogens and behavior of hygiene indicator bacteria were examined. Whereas nitrite had disappeared during the conventional packaging process, the reduced processing time in the new system allowed for the ham to be vacuum packed with retention of the nitrite (6.9±1.2 ppm, P<0.01). This accounts for the prominent decrease in L. monocytogenes (2.3 log reduction in 35 days) and S. aureus (3.3 log reduction in 13 days) counts during storage. E. coli O157 and Salmonella Enteritidis were likely resistant to the nitrite in the ham. However, they were unable to multiply in the ham and decreased gradually as in the conventionally produced ham. The bacteriostatic nature of the raw ham was also indicated by the gradual decrease in coliforms (1.3 log reduction in 13 days) in nonspiked ham. In conclusion, the raw ham produced using presliced pork loin is practically as safe as conventionally produced raw ham. It is worth validating these results in a small-scale production setting. 相似文献
855.
Hiroyuki Nagai Masaaki Kawaguchi Keita Komiya Yoshio Kamiya Masakazu Washio Shigeru Kashiwagi 《Nuclear instruments & methods in physics research. Section B, Beam interactions with materials and atoms》2007,265(1):82-86
A compact pico-second pulse radiolysis system has been developing at Waseda University for studying primary processes in radiation chemistry. The system is composed of a photo-injector system and a pico-second all-solid-state laser system. An infrared (IR) and an ultraviolet (UV) laser pulses are obtained from mode-locked Nd:YLF laser system and used for generation of the white light continuum as a probe light and the irradiation to the Cu cathode of a photo-cathode RF-gun, respectively. To improve signal-to-noise (S/N) ratio and time resolution of this pulse radiolysis system, we optimized both probe light and pump electron beam. As a result, our pico-second pulse radiolysis system has been enough to study the primary processes of radiation chemistry. The experimental results and the improvements of our system are described in this paper. 相似文献
856.
Akihiko Terada Sheng Zhou Masaaki Hosomi 《Clean Technologies and Environmental Policy》2011,13(6):759-781
Until now, anaerobic ammonium oxidation (anammox) has been widely applied as an alternative method to the conventional nitrification–denitrification
pathway for biological nitrogen removal from wastewater. Since their discovery in a denitrifying fluidized bed reactor in
the Netherlands in the early 1990s, anammox bacteria have also been detected in natural environments. Anammox is one of the
newly found drivers known to contribute to the biogeochemical nitrogen cycle. In the marine environment, more than 50% of
nitrogenous compounds are reportedly converted into nitrogen gas via the anammox pathway. These observations were made using
state-of-the-art techniques for detecting anammox bacteria based on their lipids, small-subunit ribosomal RNA genes, functional
genes, and unique reaction pathways. The research objectives for anammox bacteria are quite diverse, ranging from the application
of anammox processes to various wastewater types, to anammox biochemistry and phylogeny, to elucidating how anammox bacteria
have evolved. Since the genome of the anammox bacterium Kuenenia stuttgaritiensis was deciphered, anammox bacteria have proved to be quite versatile. The next challenge is to enrich knowledge of anammox
bacterial physiology and phylogeny to improve their use in engineered and natural environmental systems and minimize nitrogen
loads to downstream water bodies. Furthermore, rapid startup of the anammox process for engineered systems is required to
broadly harness the benefits of anammox bacteria. This review article summarizes the physiology and phylogeny of anammox bacteria,
detection methods of anammox bacteria and reactions, the behavior of anammox bacteria in natural environments, and recent
developments in their use for engineered systems. 相似文献
857.
Hitoshi Iwamoto Masaaki Okihara Isao Akashi Yu Kihara Osamu Konno Shigeyuki Kawachi Makoto Sunamura Masahiro Sugimoto 《International journal of molecular sciences》2022,23(22)
Kidney biopsy is commonly used to diagnose kidney transplant dysfunction after transplantation. Therefore, the development of minimally invasive and quantitative methods to evaluate kidney function in transplant recipients is necessary. Here, we used capillary electrophoresis-mass spectrometry to analyze the biofluids collected from transplant recipients with impaired (Group I, n = 31) and stable (Group S, n = 19) kidney function and from donors (Group D, n = 9). Metabolomics analyses identified and quantified 97 metabolites in plasma, 133 metabolites in urine, and 108 metabolites in saliva. Multivariate analyses revealed apparent differences in the metabolomic profiles of the three groups. In plasma samples, arginine biosynthesis and purine metabolism between the I and S Groups differed. In addition, considerable differences in metabolomic profiles were observed between samples collected from participants with T cell-mediated rejection (TCR), antibody-mediated rejection, and other kidney disorders (KD). The metabolomic profiles in the three types of biofluids showed different patterns between TCR and KD, wherein 3-indoxyl sulfate showed a significant increase in TCR consistently in both plasma and urine samples. These results suggest that each biofluid has different metabolite features to evaluate kidney function after transplantation and that 3-indoxyl sulfate could predict acute rejection. 相似文献
858.
859.
Nguyen Chi Trung Ngo Hironari Sugiyama Buddhika Amila Kumara Sodige Juan Paulo Wiff Satoru Yamanaka Yoonho Kim Tsuneo Suzuki Masaaki Baba Masatoshi Takeda Noboru Yamada Koichi Niihara Tadachika Nakayama 《Journal of the American Ceramic Society》2023,106(1):201-212
The energy-harvesting ability of the lead-free ferroelectric Ba(Zr,Ti)O3 was investigated and greatly enhanced using the Kim novel electrothermodynamic cycle for low-temperature application. Ba(Zr,Ti)O3 was synthesized with a Zr:Ti ratio of 10:90 (BZT10) by hot-press sintering, which exhibited a mix relaxor-ferroelectric behavior. For power generation using the Kim cycle with low and high temperatures of TL = 25°C, TH = 120°C, the most optimized temperature pattern occurred for a heating time of 12.5 s and a cooling time of 22.5 s. Under these conditions, the electric field increased during the novel isodisplacement process, and the displacement variation in the isoelectric step reached the highest value and maximized the BZT10 cycle loop area. Applying these conditions while lowering TL to 20°C, an energy density ND = 504 mJ/cm3 was achieved. This value is the highest obtained energy density in a practical test for lead-free ferroelectric bulk material in the BaTiO3 family. 相似文献
860.
Takahashi Jun Ishikawa Kyohhei Kawakami Kazuto Fujioka Masaaki 《Metallurgical and Materials Transactions A》2023,54(2):526-536
Metallurgical and Materials Transactions A - To elucidate the hardenability improvement mechanisms and hardenability-controlling factors in low-carbon (C) steels with the combined addition of... 相似文献