全文获取类型
收费全文 | 5506篇 |
免费 | 434篇 |
国内免费 | 288篇 |
专业分类
6228篇 |
出版年
2024年 | 6篇 |
2023年 | 70篇 |
2022年 | 153篇 |
2021年 | 266篇 |
2020年 | 187篇 |
2019年 | 217篇 |
2018年 | 192篇 |
2017年 | 137篇 |
2016年 | 212篇 |
2015年 | 353篇 |
2014年 | 388篇 |
2013年 | 419篇 |
2012年 | 507篇 |
2011年 | 447篇 |
2010年 | 272篇 |
2009年 | 244篇 |
2008年 | 269篇 |
2007年 | 245篇 |
2006年 | 191篇 |
2005年 | 175篇 |
2004年 | 166篇 |
2003年 | 120篇 |
2002年 | 108篇 |
2001年 | 106篇 |
2000年 | 93篇 |
1999年 | 99篇 |
1998年 | 40篇 |
1997年 | 52篇 |
1996年 | 41篇 |
1995年 | 38篇 |
1994年 | 29篇 |
1993年 | 33篇 |
1992年 | 41篇 |
1991年 | 37篇 |
1990年 | 46篇 |
1989年 | 37篇 |
1988年 | 23篇 |
1987年 | 24篇 |
1986年 | 13篇 |
1985年 | 14篇 |
1984年 | 10篇 |
1983年 | 10篇 |
1982年 | 9篇 |
1981年 | 8篇 |
1979年 | 14篇 |
1978年 | 8篇 |
1973年 | 9篇 |
1972年 | 6篇 |
1971年 | 9篇 |
1968年 | 5篇 |
排序方式: 共有6228条查询结果,搜索用时 0 毫秒
991.
A new one-dimensional manganese(III) Schiff-base complex [Mn(III)(salophen) (MeOTCNQ)] · CH3CN 1 (salophen = N,N′-bis(salicylidene)phenylenediamine) bridged by 7-methoxy-7,7,8,8-tetra-cyano-p-quinodimethane (MeOTCNQ), has been synthesized and characterized by X-ray crystallography and magnetic studies. Crystal structure study reveals that complex 1 has a 1D manganese(III) chain bridged by MeOTCNQ ligand which was obtained unexpectedly from tetracyano-p-quinodimethane (TCNQ) reacting with methanol. Noticeably, MeOTCNQ molecules in complex 1 adopt an unusual cis-syn coordination mode. The analysis of magnetic data indicates that a weak intrachain antiferromagnetic interaction exists in complex 1. 相似文献
992.
The aim of the present study is to explore whether membrane targeting of K+ channel-interacting protein 1 (KChIP1) is associated with its EF-hand motifs and varies with specific phospholipids. Truncated
KChIP1, in which the EFhands 3 and 4 were deleted, retained the α-helix structure, indicating that the N-terminal half of KChIP1 could fold appropriately. Compared with wild-type KChIP1,
truncated KChIP1 exhibited lower lipid-binding capability. Compared with wild-type KChIP1, increasing membrane permeability
by the use of digitonin caused a marked loss of truncated KChIP1, suggesting that intact EF-hands 3 and 4 were crucial for
the anchorage of KChIP1 on membrane. KChIP1 showed a higher binding capability with phosphatidylserine (PS) than truncated
KChIP1. Unlike that of truncated KChIP1, the binding of wild-type KChIP1 with membrane was enhanced by increasing the PS content.
Moreover, the binding of KChIP1 with phospholipid vesicles induced a change in the structure of KChIP1 in the presence of
PS. Taken together, our data suggest that EF-hands 3 and 4 of KChIP1 are functionally involved in a specific association with
PS on the membrane. 相似文献
993.
Role and relationship of nitric oxide and hydrogen peroxide in adventitious root development of marigold 总被引:1,自引:0,他引:1
Weibiao?Liao Honglang?Xiao Meiling?ZhangEmail author 《Acta Physiologiae Plantarum》2009,31(6):1279-1289
Several lines of evidence suggest that nitric oxide (NO) and hydrogen peroxide (H2O2) are important signal molecules involved in plant development and other physiological processes. Marigold (Tagetes erecta
L. ‘Marvel’) was used to understand the role and relationship of NO and H2O2 in adventitious root development of plants. The results showed that the effects of H2O2 or NO on adventitious root organogenesis of explants were dose dependent, with maximal biological responses at 200 μM H2O2 or 50 μM NO donor sodium nitroprusside (SNP). The results also indicated the importance of both putative NO synthase (NOS)-like
and nitrate reductase (NR) enzymes, which might be responsible for the production of NO in explants during rooting. Additionally,
guanosine 3′, 5′ -cyclic monophosphate (cGMP) was involved in NO- induced root formation of marigold, but it was not involved
in H2O2- mediated rooting process. The root number and length of explants treated with NO and H2O2 simultaneously were significantly higher than those of explants treated with H2O2 or NO alone. Moreover, NO treatments enhanced endogenous H2O2 levels in hypocotyls. Together, these results indicate that NO and H2O2 play crucial roles in the adventitious root development of marigold explants both synergistically and independently. 相似文献
994.
Our previous work has indicated that an ethylene-responsive aquaporin gene, Rh-PIP2;1, played an important role in the epidermal cell expansion of rose petals. In this work, we isolated an 896 bp promoter sequence
of the Rh-PIP2;1 and found that the promoter was rare in plants, occurring with an Inr motif, but without a TATA box. In transgenic Arabidopsis harboring the Rh-PIP2;1 promoter::GUS construct, the activity of Rh-PIP2;1 promoter was found to be developmental-dependent in almost all of the tested organs, and was particularly active in organs
that were rapidly expanding, and in tissues with high water flux capacity. Moreover, the promoter activity was inhibited by
ACC, ABA, NaCl, and cold in the roots of 3 or 6-day-old plants, and was increased by GA3 and mannitol in the rosettes of 9 or 12-day-old plants. Deleting the fragment from −886 to −828 resulted in nearly complete
disappearance of the promoter activity in roots, and a substantial decrease in the leaves, hypocotyls and floral organs. Taken
together, our results indicated that the Rh-PIP2;1 promoter responded to hormones and abiotic stresses in a developmental- and spatial-dependent manner, and the −886 to −828
region was crucial for the activity of the Rh-PIP2;1 promoter.
Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.
Y. Li and Z. Wu contributed to this work equally. 相似文献
995.
Cardiomyocyte death caused by proinflammatory cytokines, such as Tumor necrosis factor α (TNF-α), is one of the hot topics in cardiovascular research. TNF-α can induce multiple cell processes that are dependent on the treatment time although the long-term treatment definitely leads to cell death. The ability to intervene in cell death will be invaluable to reveal the effects of short-term TNF-α treatment to cardiomyocytes. However, a real-time monitoring technique is needed to guide the intervention of cell responses. In this work, we employed the impedance-sensing technique to real-time monitor the equivalent cell–substrate distance of cardiomyocytes via electrochemical impedance spectroscopy (EIS) and electrical cell–substrate impedance sensing (ECIS). In the stabilized cardiomyocyte culture, the sustained TNF-α treatment caused strengthened cell adhesion in the first 2 h which was followed by the transition to cell detachment afterwards. Considering cell detachment was an early morphological evidence of cell death, we removed TNF-α from the cardiomyocyte culture before the transition to achieve the intervention of cell responses. The result of this intervention showed that cell adhesion was continuously strengthened before and after the removal of TNF-α, indicating the short-term treated cardiomyocytes did not undergo death processes. It was also demonstrated in TUNEL and TBE tests that the percentages of apoptosis and cell death were both lowered. 相似文献
996.
Hui‐Min Wang Yan‐Ping Shih Su‐Ming Hu Wen‐Tsung Lo Hui‐Min Lin Shih‐Shiu Ding Hsin‐Chi Liao Po‐Huang Liang 《Biotechnology progress》2009,25(6):1582-1586
To quickly find an optimal expression system for recombinant protein production, a set of vectors with the same restriction sites were constructed for parallel cloning of a target gene and recombinant protein production in prokaryotic and eukaryotic expression systems, simultaneously. These vectors include nucleotide sequences encoding protein tags and protease recognition sites for tag removal, followed by the cloning sites 5′‐EcoRI/3′‐XhoI identical in these vectors for ligating with the sticky‐end PCR product of a target gene. Our vectors allow parallel gene cloning and protein production in multiple expression systems with minimal cloning effort. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2009 相似文献
997.
椒江口滩涂大型底栖动物群落格局与多样性 总被引:6,自引:0,他引:6
为了解椒江口滩涂大型底栖动物群落格局与多样性, 揭示其对环境变化的响应规律, 作者于2007年10月、2008年1月、4月和7月在椒江口南岸和北岸潮间带, 沿河流到海洋方向共布设6条采样断面进行大型底栖动物调查。分析了大型底栖动物种类组成、栖息密度和生物量的时空变化特征, 在此基础上运用α, β和γ多样性测度方法对大型底栖动物多样性进行分析, 同时探讨了大型底栖动物群落结构对环境变化的响应方向及程度, 结果显示: (1) 6条断面共记录到大型底栖动物78种, 总种数随季节变化显著, 在空间上沿河流到海洋方向呈升高趋势; (2) 栖息密度的季节变化不显著(P=0.145>0.05), 但空间变化显著(P=0.017<0.05), 生物量的季节变化显著(P=0.012<0.05), 空间变化极显著(P=0.004<0.01); (3) β和γ多样性指数定量显示了椒江河口区域滩涂环境的多变性和大型底栖动物群落的多样性和更替性。 相似文献
998.
999.
Kuo-Liang Su Ya-Fan Liao Hui-Chih Hung Guang-Yaw Liu 《The Journal of biological chemistry》2009,284(39):26768-26777
Ornithine decarboxylase (ODC) is the first enzyme involved in polyamine biosynthesis, and it catalyzes the decarboxylation of ornithine to putrescine. ODC is a dimeric enzyme, whereas antizyme inhibitor (AZI), a positive regulator of ODC that is homologous to ODC, exists predominantly as a monomer and lacks decarboxylase activity. The goal of this paper was to identify the essential amino acid residues that determine the dimerization of AZI. The nonconserved amino acid residues in the putative dimer interface of AZI (Ser-277, Ser-331, Glu-332, and Asp-389) were substituted with the corresponding residues in the putative dimer interface of ODC (Arg-277, Tyr-331, Asp-332, and Tyr-389, respectively). Analytical ultracentrifugation analysis was used to determine the size distribution of these AZI mutants. The size-distribution analysis data suggest that residue 331 may play a major role in the dimerization of AZI. Mutating Ser-331 to Tyr in AZI (AZI-S331Y) caused a shift from a monomer configuration to a dimer. Furthermore, in comparison with the single mutant AZI-S331Y, the AZI-S331Y/D389Y double mutant displayed a further reduction in the monomer-dimer Kd, suggesting that residue 389 is also crucial for AZI dimerization. Analysis of the triple mutant AZI-S331Y/D389Y/S277R showed that it formed a stable dimer (Kd value = 1.3 μm). Finally, a quadruple mutant, S331Y/D389Y/S277R/E332D, behaved as a dimer with a Kd value of ∼0.1 μm, which is very close to that of the human ODC enzyme. The quadruple mutant, although forming a dimer, could still be disrupted by antizyme (AZ), further forming a heterodimer, and it could rescue the AZ-inhibited ODC activity, suggesting that the AZ-binding ability of the AZI dimer was retained.Polyamines (putrescine, spermidine, and spermine) have been shown to have both structural and regulatory roles in protein and nucleic acid biosynthesis and function (1–3). Ornithine decarboxylase (ODC,3 EC 4.1.1.17) is a central regulator of cellular polyamine synthesis (reviewed in Refs. 1, 4, 5). This enzyme catalyzes the pyridoxal 5-phosphate (PLP)-dependent decarboxylation of ornithine to putrescine, and it is the first and rate-limiting enzyme in polyamine biosynthesis (2, 3, 6, 7). ODC and polyamines play important roles in a number of biological functions, including embryonic development, cell cycle, proliferation, differentiation, and apoptosis (8–15). They also have been associated with human diseases and a variety of cancers (16–26). Because the regulation of ODC and polyamine content is critical to cell proliferation (11), as well as in the origin and progression of neoplastic diseases (23, 24), ODC has been identified as an oncogenic enzyme, and the inhibitors of ODC and the polyamine pathway are important targets for therapeutic intervention in many cancers (6, 11).ODC is ubiquitously found in organisms ranging from bacteria to humans. It contains 461 amino acid residues in each monomer and is a 106-kDa homodimer with molecular 2-fold symmetry (27, 28). Importantly, ODC activity requires the formation of a dimer (29–31). X-ray structures of the ODC enzyme reveal that this dimer contains two active sites, both of which are formed at the interface between the N-terminal domain of one monomer, which provides residues involved in PLP interactions, and the C-terminal domain of the other subunit, which provides the residues that interact with substrate (27, 32–41).ODC undergoes a unique ubiquitin-independent proteasomal degradation via a direct interaction with the regulatory protein antizyme (AZ). Binding of AZ promotes the dissociation of the ODC homodimers and targets ODC for degradation by the 26 S proteasome (42–46). Current models of antizyme function indicate that increased polyamine levels promote the fidelity of the AZ mRNA translational frameshift, leading to increased concentrations of AZ (47). The AZ monomer selectively binds to dimeric ODC, thereby inactivating ODC by forming inactive AZ-ODC heterodimers (44, 48–50). AZ acts as a regulator of polyamine metabolism that inhibits ODC activity and polyamine transport, thus restricting polyamine levels (4, 5, 51, 52). When antizymes are overexpressed, they inhibit ODC and promote ubiquitin-independent proteolytic degradation of ODC. Because elevated ODC activity is associated with most forms of human malignancies (1), it has been suggested that antizymes may function as tumor suppressors.In contrast to the extensive studies on the oncogene ODC, the endogenous antizyme inhibitor (AZI) is less well understood. AZI is homologous to the enzyme ODC. It is a 448-amino acid protein with a molecular mass of 50 kDa. However, despite the homology between these proteins, AZI does not possess any decarboxylase activity. It binds to antizyme more tightly than does ODC and releases ODC from the ODC-antizyme complex (53, 54). Both the AZI and AZ proteins display rapid ubiquitin-dependent turnover within a few minutes to 1 h in vivo (5). However, AZ binding actually stabilizes AZI by inhibiting its ubiquitination (55).AZI, which inactivates all members of the AZ family (53, 56), restores ODC activity (54), and prevents the proteolytic degradation of ODC, may play a role in tumor progression. It has been reported that down-regulation of AZI is associated with the inhibition of cell proliferation and reduced ODC activity, presumably through the modulation of AZ function (57). Moreover, overexpression of AZI has been shown to increase cell proliferation and promote cell transformation (58–60). Furthermore, AZI is capable of direct interaction with cyclin D1, preventing its degradation, and this effect is at least partially independent of AZ function (60, 61). These results demonstrate a role for AZI in the positive regulation of cell proliferation and tumorigenesis.It is now known that ODC exists as a dimer and that AZI may exist as a monomer physiologically (62). Fig. 1 shows the dimeric structures of ODC (Fig. 1A) and AZI (Fig. 1B). Although structural studies indicate that both ODC and AZI crystallize as dimers, the dimeric AZI structure has fewer interactions at the dimer interface, a smaller buried surface area, and a lack of symmetry of the interactions between residues from the two monomers, suggesting that the AZI dimer may be nonphysiological (62). In this study, we identify the critical amino acid residues governing the difference in dimer formation between ODC and AZI. Our preliminary studies using analytical ultracentrifugation indicated that ODC exists as a dimer, whereas AZI exists in a concentration-dependent monomer-dimer equilibrium. Multiple sequence alignments of ODC and AZI from various species have shown that residues 277, 331, 332, and 389 are not conserved between ODC and AZI (Open in a separate windowFIGURE 1.Crystal structure and the amino acid residues at the dimer interface of human ornithine decarboxylase (hODC) and mouse antizyme inhibitor (mAZI). A, homodimeric structure of human ODC with the cofactor PLP analog, LLP (Protein Data Bank code 1D7K). B, putative dimeric structure of mouse AZI (Protein Data Bank code 3BTN). The amino acid residues in the dimer interface are shown as a ball-and-stick model. The putative AZ-binding site is colored in cyan. This figure was generated using PyMOL (DeLano Scientific LLC, San Carlos, CA).
Open in a separate window 相似文献
TABLE 1
Amino acid residues at the dimer interface of human ODC and AZIHuman ODC | Residue | Human AZI |
---|---|---|
Nonconserved | ||
Arg | 277 | Ser |
Tyr | 331 | Ser |
Asp | 332 | Glu |
Tyr | 389 | Asp |
Conserved | ||
Asp | 134 | Asp |
Lys | 169 | Lys |
Lys | 294 | Lys |
Tyr | 323 | Tyr |
Asp | 364 | Asp |
Gly | 387 | Gly |
Phe | 397 | Phe |