首页 | 本学科首页   官方微博 | 高级检索  
文章检索
  按 检索   检索词:      
出版年份:   被引次数:   他引次数: 提示:输入*表示无穷大
  收费全文   5576篇
  免费   192篇
  国内免费   1篇
  5769篇
  2022年   27篇
  2021年   52篇
  2020年   23篇
  2019年   51篇
  2018年   58篇
  2017年   39篇
  2016年   103篇
  2015年   167篇
  2014年   183篇
  2013年   377篇
  2012年   284篇
  2011年   261篇
  2010年   182篇
  2009年   209篇
  2008年   295篇
  2007年   286篇
  2006年   279篇
  2005年   293篇
  2004年   302篇
  2003年   294篇
  2002年   294篇
  2001年   114篇
  2000年   129篇
  1999年   104篇
  1998年   72篇
  1997年   69篇
  1996年   62篇
  1995年   49篇
  1994年   53篇
  1993年   64篇
  1992年   92篇
  1991年   75篇
  1990年   70篇
  1989年   58篇
  1988年   62篇
  1987年   58篇
  1986年   55篇
  1985年   49篇
  1984年   45篇
  1983年   38篇
  1982年   37篇
  1981年   41篇
  1980年   28篇
  1979年   29篇
  1978年   37篇
  1976年   23篇
  1974年   19篇
  1973年   17篇
  1972年   18篇
  1969年   17篇
排序方式: 共有5769条查询结果,搜索用时 15 毫秒
1.
Heterodichogamy is a form of sex expression in which protandrous and protogynous individuals coexist, and is considered to be a mechanism that avoids selfing and promotes disassortative mating. We examined mating patterns in a heterodichogamous maple, Acer mono, using microsatellite markers. Parentage analysis revealed a selfing rate of only 9.8%. Disassortative mating between flowering types significantly exceeded within-type mating, but the mating patterns were better explained by flowering phenology (i.e., the temporal overlap between the female and male stages). Heterodichogamy in A. mono thus appears to promote outcrossing without requiring obligate self- or cross-incompatibility systems, although it did not guarantee disassortative mating. Multiple-regression analysis suggested that successful reproduction of pollen parents significantly increased with increased flower production and reciprocal flowering synchrony, but decreased only marginally with mating distance, although the distribution of mating distances suggested leptokurtic dispersal of pollen.  相似文献   
2.
A high-performance liquid chromatographic assay was developed for the quantitative determination of the sulfur-containing amino acids N-acetyl- -cysteine (NAC) and -cysteine (Cys) in rat plasma. The thiols were separated by reverse-phase ion-pair chromatography, and the column eluent was continuously mixed with an iodoplatinate-containing solution. The substitution of sulfur of the thiol compound with iodide was quantitatively determined by measuring changes in the absorption at 500 nm. The low-molecular-weight disulfides and mixed disulfide conjugates of thiols with proteins were entirely reduced to the original reduced compounds by dithiothreitol. By reducing these two types of disulfides separately during sample pretreatment, the reduced, protein-unbound, and total thiol concentrations could also be determined. Validation testing was performed, and no problems were encountered. The limit of detection was approximately 20 pmol of thiol on the column. The present method was used to measure the plasma concentrations of NAC and Cys in the rat after a bolus intravenous administration of NAC, focusing on disulfide formation. The binding of NAC to protein through mixed disulfide formation proceeds in a time-dependent and reversible manner. Moreover, this “stable” covalent binding might limit total drug elimination, while the unbound NAC is rapidly eliminated. Consequently, the analytical method described in this study is very useful for the determination of plasma NAC and Cys, including disulfide conjugates derived from them.  相似文献   
3.
Visible absorption and CD spectral and potentiometric studies on the His- and Tyr-containing ternary copper(II) complexes Cu(A)(L-B), where A refers to L-His, D-His, or L-Tyr and B to Lys, Tyr, Trp, Phe, Ala, Val, Arg, Glu, Asn, Gln, Ser, or Thr, were made to study ligand-ligand interactions in the complexes. While the CD spectral magnitudes in the d—d region are additive in the absence of side chain interactions and can be estimated from the magnitudes for the ternary systems involving DL-A or DL-B, deviation from the additivity was observed for Cu(L-His)(L-B) (B = LysH, Tyr, Trp, or Phe) and Cu(L-Tyr)(L-Trp). From the stability constants determined at 25 °C and I = 0.1 M (KNO3), the equilibrium constants, K, for the following hypothetical equilibria were calculated to be large (0.14–0.60) for formation of Cu(L-/D-His)(L-B)(B = Tyr or Trp) and Cu(D-His)(L-Phe) with Cu(en)(L-Ala) as standard: Cu(A)(L?Ala)+Cu(en)(L?b)?KCu(A)(L?B)+Cu(en)(L?Ala) The positive values indicate the stabilization due to the stacking between the imidazole ring of His and the aromatic side chain of L-B. Solvent dependence of the CD spectra for Cu(L-His)(L-LysH) and Cu(L-His) L-Trp) further supported the existence of the intramolecular electrostatic and hydrophobic interactions.  相似文献   
4.
To elucidate the exact role of the PGD(2) system in sleep-wake regulation in vivo, the sleep behavior of knockout mice, generated in the author's and other laboratories, was examined for lipocalin-type PGD synthase (L-PGDS), PGD receptor, adenosine A(2A) receptor, and histamine H(1) receptor; transgenic mice overexpressing the human L-PGDS gene, generated in the author's laboratory, were also examined. The circadian profiles of sleep patterns of wild-type and the genetically manipulated mice were essentially identical, indicating the possibility that the deficiency of one system may be effectively compensated by some other systems during development. Available evidence indicated that the PGD(2) system is involved in the homeostatic regulation of non-rapid eye movement sleep and that the arousal effect of orexin A is mediated by the histamine H(1) receptor system.  相似文献   
5.
6.

Background, aim, and scope  

To achieve sustainable development in industrial processes, attributed chemical risks as well as environmental impacts should be managed. Such non-monetary issues have been analyzed by scientific assessment methodologies such as various risk assessment (RA) and life cycle assessment (LCA) procedures. Local risks to be addressed in RA are microenvironments, including the workplace and neighborhood. Although a comprehensive interpretation of such risks is necessitated in industrial decision making, no practical method has been developed to interpret various types of risk with sufficient understandings of plant-specific functions and constraints. Because elaborate model-based approaches are inevitable for practical process development, actual case studies on chemical risks and detailed plant-specific functions and constraints should be performed. Manufacturing processes require that metal parts must be cleaned in preparation for surface treatments or the completion of metal processing. The significant amount of cleansing agents utilized in cleaning processes has become an issue in Japan. Almost all cleaning processes in Japan are carried out by small- and medium-sized enterprises (SMEs). Machinery processes have not been systematically analyzed in terms of chemical risks and, in addition, the environmental management skills of SMEs are generally far behind those of large enterprises. The objective of this study is to reveal the relationships between chemical risks and plant-specific conditions for a practical risk reduction carried out by industrial decision makers. For this purpose, we aimed at the analysis of such relationships in metal-cleaning processes. Through this analysis, the correlation between local risks and global impacts were discussed in terms of plant-specific conditions.  相似文献   
7.
The molecular basis of the genetic code relies on the specific ligation of amino acids to their cognate tRNA molecules. However, two pathways exist for the formation of Gln-tRNAGln. The evolutionarily older indirect route utilizes a non-discriminating glutamyl-tRNA synthetase (ND-GluRS) that can form both Glu-tRNAGlu and Glu-tRNAGln. The Glu-tRNAGln is then converted to Gln-tRNAGln by an amidotransferase. Since the well-characterized bacterial ND-GluRS enzymes recognize tRNAGlu and tRNAGln with an unrelated α-helical cage domain in contrast to the β-barrel anticodon-binding domain in archaeal and eukaryotic GluRSs, the mode of tRNAGlu/tRNAGln discrimination in archaea and eukaryotes was unknown. Here, we present the crystal structure of the Methanothermobacter thermautotrophicus ND-GluRS, which is the evolutionary predecessor of both the glutaminyl-tRNA synthetase (GlnRS) and the eukaryotic discriminating GluRS. Comparison with the previously solved structure of the Escherichia coli GlnRS-tRNAGln complex reveals the structural determinants responsible for specific tRNAGln recognition by GlnRS compared to promiscuous recognition of both tRNAs by the ND-GluRS. The structure also shows the amino acid recognition pocket of GluRS is more variable than that found in GlnRS. Phylogenetic analysis is used to reconstruct the key events in the evolution from indirect to direct genetic encoding of glutamine.  相似文献   
8.
D-Aminoacylase from Alcaligenes xylosoxydans subsp. xylosoxydans A-6 (AxD-NAase) offers a novel biotechnological application, the production of D-amino acid from the racemic mixture of N-acyl-DL-amino acids. However, its substrate specificity is biased toward certain N-acyl-D-amino acids. To construct mutant AxD-NAases with substrate specificities different from those of wild-type enzyme, the substrate recognition site of the AxD-NAase was rationally manipulated based on computational structural analysis and comparison of its primary structure with other D-aminoacylases with distinct substrate specificities. Mutations of amino acid residues, Phe191, Leu298, Tyr344, and Met346, which interact with the side chain of the substrate, induced marked changes in activities toward each substrate. For example, the catalytic efficiency (k(cat)/K(m)) of mutant F191W toward N-acetyl-D-Trp and N-acetyl-D-Ala was enhanced by 15.6- and 1.5-folds, respectively, compared with that of the wild-type enzyme, and the catalytic efficiency (k(cat)/K(m)) of mutant L298A toward N-acetyl-D-Trp was enhanced by 4.4-folds compared with that of the wild-type enzyme. Other enzymatic properties of both mutants, such as pH and temperature dependence, were the same as those of the wild-type enzyme. The F191W mutant in particular is considered to be useful for the enzymatic production of D-Trp which is an important building block of some therapeutic drugs.  相似文献   
9.
A nucleosome assembly protein (NAP-1) of Saccharomyces cerevisiae facilitates the association of histones with DNA to form nucleosomes in vitro at physiological ionic conditions. The cloned gene was expressed in Escherichia coli using a T7 expression system, and the protein (417 amino acid residues) was purified by Mono Q column chromatography. Various deletion fragments of NAP-1 protein were also produced, and their nucleosome assembly activity was examined by supercoiling assay. The internal fragment containing the residues 43-365 was necessary and sufficient for the activity, and a long stretch of negatively charged region near the carboxyl terminus was dispensable. This minimal size fragment could form the 12 S NAP-1-histone complex as the whole protein could, whereas deleted fragments on either side could bind with core histones only to form aggregates.  相似文献   
10.
SGIP1 has been shown to be an endophilin-interacting protein that regulates energy balance, but its function is not fully understood. Here, we identified its splicing variant of SGIP1 and named it SGIP1alpha. SGIP1alpha bound to phosphatidylserine and phosphoinositides and deformed the plasma membrane and liposomes into narrow tubules, suggesting the involvement in vesicle formation during endocytosis. SGIP1alpha furthermore bound to Eps15, an important adaptor protein of clathrin-mediated endocytic machinery. SGIP1alpha was colocalized with Eps15 and the AP-2 complex. Upon epidermal growth factor (EGF) stimulation, SGIP1alpha was colocalized with EGF at the plasma membrane, indicating the localization of SGIP1alpha at clathrin-coated pits/vesicles. SGIP1alpha overexpression reduced transferrin and EGF endocytosis. SGIP1alpha knockdown reduced transferrin endocytosis but not EGF endocytosis; this difference may be due to the presence of redundant pathways in EGF endocytosis. These results suggest that SGIP1alpha plays an essential role in clathrin-mediated endocytosis by interacting with phospholipids and Eps15.  相似文献   
设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号