全文获取类型
收费全文 | 8121篇 |
免费 | 537篇 |
国内免费 | 1294篇 |
出版年
2023年 | 129篇 |
2022年 | 208篇 |
2021年 | 281篇 |
2020年 | 256篇 |
2019年 | 284篇 |
2018年 | 248篇 |
2017年 | 273篇 |
2016年 | 337篇 |
2015年 | 335篇 |
2014年 | 385篇 |
2013年 | 732篇 |
2012年 | 311篇 |
2011年 | 354篇 |
2010年 | 281篇 |
2009年 | 377篇 |
2008年 | 350篇 |
2007年 | 371篇 |
2006年 | 380篇 |
2005年 | 313篇 |
2004年 | 320篇 |
2003年 | 302篇 |
2002年 | 281篇 |
2001年 | 193篇 |
2000年 | 172篇 |
1999年 | 166篇 |
1998年 | 153篇 |
1997年 | 154篇 |
1996年 | 123篇 |
1995年 | 141篇 |
1994年 | 160篇 |
1993年 | 151篇 |
1992年 | 132篇 |
1991年 | 94篇 |
1990年 | 81篇 |
1989年 | 113篇 |
1988年 | 76篇 |
1987年 | 73篇 |
1986年 | 65篇 |
1985年 | 106篇 |
1984年 | 147篇 |
1983年 | 97篇 |
1982年 | 100篇 |
1981年 | 64篇 |
1980年 | 51篇 |
1979年 | 57篇 |
1978年 | 40篇 |
1977年 | 24篇 |
1976年 | 33篇 |
1974年 | 26篇 |
1973年 | 20篇 |
排序方式: 共有9952条查询结果,搜索用时 15 毫秒
991.
AIM:To determine if the cytotail of the principal sheddase tumor necrosis factor-α converting enzyme (TACE;ADAM17) controls protein ectodomain shedding.METHODS:Site-directed mutagenesis was performed to derive TACE variants. The resulting TACE expression plasmids with amino acid substitutions in the extracel-lular,cysteine-rich disintegrin domain (CRD) and/or deleted cytotail,along with an expression vector for the enhanced green fluorescence protein were transfected into shedding-defective M1 mutants stably expressing transmembrane L-selectin or transforming growth factor (TGF)-α. The expression levels of the TACE substrates at the cell surface were determined by flow cytometry. RESULTS:Consistent with published data,a single point mutation (C600Y) in the CRD led to shedding defi-ciency. However,removal of the cytotail from the C600Y TACE variant partially restored ectodomain cleavage of TGF-α and L-selectin. Cytotail-deleted mutants with any other substituting amino acid residues in place of Cys600 displayed similar function compared with tail-less C600Y TACE.CONCLUSION:The cytotail plays an inhibitory role,which becomes evident when it is removed from an enzyme with another mutation that affects the enzyme function. 相似文献
992.
Volpato G Filice M de las Rivas B Rodrigues RC Heck JX Fernandez-Lafuente R Guisan JM Mateo C Ayub MA 《Biotechnology progress》2011,27(3):717-723
Staphylococcus warneri strain EX17 produces three lipases with different molecular weights of 28, 30, and 45 kDa. The 45 kDa fraction (SWL-45) has been purified from crude protein extracts by one chromatographic step based on the selective adsorption of this lipase by interfacial activation on different hydrophobic supports at low ionic strength. The adsorption of SWL-45 on octyl-Sepharose increased the enzyme activity by 60%, but the other lipases were also adsorbed on this support. Using butyl-Toyopearl, which is a lesser hydrophobic support, the purification factor was close to 20, and the only protein band detected on the sodium dodecyl sulfate-polyacrylamide electrophoresis analysis gel was that corresponding to the SWL-45, which could be easily desorbed from the support by incubation with triton X-100, producing a purified enzyme. SWL-45 was immobilized under very mild conditions on cyanogen bromide Sepharose, showing similar activities and stability as for its soluble form but without intermolecular interaction. The effects of different detergents over the activity of the immobilized SWL-45 were analyzed, which was hyperactivated by factors of 1.3 and 2.5 with 0.01% Tween 80 and 0.1% Triton X-100, respectively, while ionic detergents produced detrimental effects on the enzyme activity even at very low concentrations. Optimal reaction conditions and the effect of other additives on the enzyme activity were also investigated. 相似文献
993.
Bibi NS Gavara PR Soto Espinosa SL Grasselli M Fernández-Lahore M 《Biotechnology progress》2011,27(5):1329-1338
The preparation of megaporous bodies, with potential applications in biotechnology, was attempted by following several strategies. As a first step, naive and robust scaffolds were produced by polymerization of selected monomers in the presence of a highly soluble cross‐linker agent. Ion‐exchange function was incorporated by particle embedding, direct chemical synthesis, or radiation‐induced grafting. The total ionic capacity of such systems was 1.5 mmol H+/g, 1.4 mmol H+/g, and 17 mmol H+/g, respectively. These values were in agreement with the ability to bind model proteins: observed dynamic binding capacity at 50% breakthrough was ?7.2 mg bovine serum albumin/g, ?7.4 hen egg‐white lysozyme (HEWL) mg/g, and ?108 HEWL mg/g. In the later case, total (static) binding capacity reached 220 mg/g. It was observed that the structure and size of the megapores remained unaffected by the grafting procedure which, however, allowed for the highest protein binding capacity. Lysozyme supported on grafted body showed extensive clarification activity against a Micrococcus lysodekticus suspension in the flow‐through mode, i.e., 90% destruction of suspended microbial cells was obtained with a residence time ≈ 18 min. Both protein capture and biocatalysis applications are conceivable with the 3D‐megaporous materials described in this work. © 2011 American Institute of Chemical Engineers Biotechnol. Prog., 2011 相似文献
994.
995.
十二烷基硫酸钠协同微波-酶法提取布渣叶总黄酮 总被引:1,自引:1,他引:0
采用十二烷基硫酸钠(SDS)协同微波-酶法提取布渣叶总黄酮,选取微波辐射时间(X1)、SDS质量分数(X2)、液固比(X3)、恒温水浴提取时间(X4)、酶解时间(X5)进行U15(54×3)混合水平均匀试验设计,利用偏最小二乘法(PLS)处理试验数据,建立数学模型,确定提取的最优条件并进行验证;另外,将优化后的提取结果与其他提取方法进行比较.研究结果表明:优化的提取条件为X1=1min、X2=0.16%、X3=30.75 mL/g、X4=50 min、X5=30 min.与其他提取方法相比,该法提取效率高,环保无污染,具有一定的应用价值. 相似文献
996.
997.
廖清池周胜华 《现代生物医学进展》2011,11(14):2779-2782
microRNA(miRNA)是一类非编码的小分子单链RNA,主要通过转录后抑制靶基因表达调节各种生物功能.miRNA表达水平在正常发育过程中以及在癌症及心血管疾病等各种疾病中发生变化.目前控制miRNA生物合成的信号途径以及调节机制尚未完全阐明.miRNA基因转录后在大蛋白复合体的介导下经一系列协调加工过程形成成熟的miRNA.近来发现转化生长因子B(TGFβ)信号途径的转导因子Smad蛋白在细胞核内调节miRNA的加工处理过程起重要作用.本文主要综述TGFB/Smad信号途径在miRNA生物合成中的调节作用. 相似文献
998.
Werner C Janisch S Wicke M 《Animal : an international journal of animal bioscience》2011,5(5):813-820
Colour is an important quality parameter of broiler meat influencing the consumer buying behaviour. The alterations of the colour after slaughter are related to the oxidative status of the tissue. This in turn is influenced by an interaction between the mitochondria and the antioxidative enzymes. In this study, breast muscles were collected from hens and cocks of a commercial line slaughtered at the ages of 28 and 41 day. Analysis of the activities of superoxide dismutase (SOD), glutathione peroxidase (GPx) and glutathione reductase (GR) was performed with samples obtained 20 min and 48 h after slaughter (post mortem, p.m.), whereas the mitochondrial respiratory activity was analysed in permeabilised breast muscle fibres collected 20 min p.m. The carcass characteristics of breast muscle and leg weight as well as breast yield were significantly higher, and the leg yields lower, in the 41-day-old broiler. The 28-day-old hens and cocks had comparable carcass characteristics (P > 0.05), whereas 41-day-old cocks had significantly higher carcass, breast and leg weight in comparison to the hens. The pH20 min p.m. and the L*48 h p.m. were significantly higher, and the a* and b* values of the 20 min and 48 h p.m. samples as well as the drip loss were significantly lower in the 41-day-old broiler. Mitochondrial respiratory rates were comparable (P > 0.05) between the 28- and 41-day-old cocks and hens. The same result could be found with regard to the activities of the SOD, GPx and GR except for lower activities of the SOD20 min p.m. and higher of the GR48 h min p.m. in the 41-day-old broiler. The concentrations of thiobarbituric acid-reactive substances were generally higher in the breast muscles of the 41-day-old broiler. Assorting the data according to their mean pH20 min p.m. indicates a positive influence of higher pH values (>6.34) on the mitochondrial function, whereas a low pH20 min p.m. results in tendentially and significantly higher activities of the antioxidative enzymes and drip loss values. These results indicate a relation between the meat quality and the oxidative metabolism as well as antioxidative capacity of the meat. 相似文献
999.
Zachary L. Rinkes Michael N. Weintraub Jared L. DeForest Daryl L. Moorhead 《Fungal Ecology》2011,4(6):396-407
The Guild Decomposition Model (GDM) hypothesized that temporal shifts in microbial “guilds,” each with distinct substrate preferences, drive decomposition dynamics and regulate soil carbon (C) losses and sequestration. To test this hypothesis, we established a laboratory incubation of Acer saccharum litter and monitored respiration, microbial biomass and enzyme activities, inorganic nutrients and shifts in functional groups of decomposers using phospholipid fatty acid (PLFA) analysis. Biomass and respiration peaked within the first 2 d of incubation, and the Gram negative PLFA biomarker 18:1ω7c predominated during the first 5 d. Hydrolytic enzyme activities and two fungal biomarkers (18:2ω6,9c and 18:3ω6c) increased by 25 d and lignolytic enzyme activity was detected at 68 d. Our results suggest that decomposers preferentially use labile substrates and that shifts in decomposer groups occur in response to changes in available substrates, which supports the GDM. 相似文献
1000.
Hydrophilins from distant organisms can protect enzymatic activities from water limitation effects in vitro 总被引:2,自引:0,他引:2