全文获取类型
收费全文 | 15449篇 |
免费 | 827篇 |
国内免费 | 8篇 |
出版年
2022年 | 76篇 |
2021年 | 123篇 |
2020年 | 64篇 |
2019年 | 98篇 |
2018年 | 153篇 |
2017年 | 161篇 |
2016年 | 264篇 |
2015年 | 421篇 |
2014年 | 469篇 |
2013年 | 1229篇 |
2012年 | 837篇 |
2011年 | 909篇 |
2010年 | 520篇 |
2009年 | 499篇 |
2008年 | 837篇 |
2007年 | 922篇 |
2006年 | 935篇 |
2005年 | 963篇 |
2004年 | 1029篇 |
2003年 | 992篇 |
2002年 | 923篇 |
2001年 | 218篇 |
2000年 | 181篇 |
1999年 | 207篇 |
1998年 | 253篇 |
1997年 | 236篇 |
1996年 | 194篇 |
1995年 | 178篇 |
1994年 | 135篇 |
1993年 | 170篇 |
1992年 | 166篇 |
1991年 | 139篇 |
1990年 | 129篇 |
1989年 | 122篇 |
1988年 | 116篇 |
1987年 | 89篇 |
1986年 | 100篇 |
1985年 | 108篇 |
1984年 | 120篇 |
1983年 | 107篇 |
1982年 | 114篇 |
1981年 | 110篇 |
1980年 | 94篇 |
1979年 | 56篇 |
1978年 | 60篇 |
1977年 | 49篇 |
1976年 | 66篇 |
1975年 | 39篇 |
1974年 | 49篇 |
1973年 | 42篇 |
排序方式: 共有10000条查询结果,搜索用时 15 毫秒
991.
As the numbers of middle-aged and elderly mountain climbers have increased with the general aging of the population, accidents during mountain climbing have increased recently. A possible cause of such accidents is an excessively difficult expedition plan. To enjoy safe mountain climbing, the plans must take account of the climber's fitness level. We developed a method to plan mountain climbing using the climbing heartbeat index (CHI). This study is based on the assumption that the work expended when climbing a mountain is equal to the potential energy of the body and load weights elevated to the height of the mountain, and that the work is proportional to the heart rate. The CHI was calculated by the following equation The CHI values examined in this study ( n = 94) showed very small standard deviations and were significantly correlated with the maximum oxygen uptake, .VO(2 max) (ml kg(-1) min(-1)) ( r = -0.934, P < 0.01); it showed a characteristic value corresponding to the fitness level in each subject. In addition, this value remained nearly unchanged even when the load was changed. Therefore, if the CHI value of an individual is known (it can be estimated from .VO(2 max)), safer mountain climbing can be planned accordingly. Once determined, this CHI value can be used repeatedly unless the fitness level of the individual changes. 相似文献
992.
Mannitol-1-phosphate dehydrogenase (MtlD) is required for mannitol and glucitol assimilation in Bacillus subtilis: possible cooperation of mtl and gut operons
下载免费PDF全文
![点击此处可从《Journal of bacteriology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
Watanabe S Hamano M Kakeshita H Bunai K Tojo S Yamaguchi H Fujita Y Wong SL Yamane K 《Journal of bacteriology》2003,185(16):4816-4824
993.
994.
Kashiwagi K Shiba K Fukami-Kobayashi K Noda T Nishikawa K Noguchi H 《Journal of biochemistry》2003,133(3):371-376
The family of periplasmic binding proteins (PBPs) is believed to have arisen from a common ancestor and to have differentiated into two types. At first approximation, both types of PBPs have the same fold pattern, reflecting their common origin. However, the connection between the main chains of a type 2 PBP is more complicated than a type 1 PBP's. We have been interested in the possibility that such structural changes affect the folding of PBPs. In this study, we have characterized the folding pathways of MglB (a type 1 PBP) and ArgT (a type 2 PBP) by using urea gradient gel electrophoresis, fast protein size-exclusion liquid chromatography and hydrophobic dye ANS binding assay. We found a distinct difference in folding between these two proteins. The folding of MglB followed a simple two-state transition model, whereas the folding of ArgT was more complicated. 相似文献
995.
Sakashita G Shima H Komatsu M Urano T Kikuchi A Kikuchi K 《Journal of biochemistry》2003,133(2):165-171
Inhibitor 2 (I-2) is a ubiquitous regulator of type 1 protein phosphatase (PP1). Previous in vitro studies suggested that its inhibitory activity towards PP1 is regulated by phosphorylation at Thr72 by glycogen synthase kinase-3beta (GSK-3beta), and at Ser86, Ser120, and Ser121 by casein kinase 2 (CK2). Here we report that GSK-3beta expressed in COS-7 cells phosphorylates wild-type I-2 but not an I-2 mutant carrying a T to A substitution at residue 72, showing that GSK-3beta phosphorylates I-2 at T72 in vivo as well. Co-immunoprecipitation study demonstrated that HA-GSK-3beta and I-2-FLAG co-exist in a same complex in the intact cells, but they do not bind directly. It is noteworthy that co-expression of Myc-PP1C significantly increased co-precipitation of HA-GSK-3beta with I-2-FLAG, showing a complex formation of HA-GSK-3beta/Myc-PP1C / I-2-FLAG in vivo. Further studies using a GSK-3beta kinase-dead mutant and LiCl, an inhibitor of GSK-3beta, showed that the enzyme activity of GSK-3beta is required for co-precipitation. IP-Western study using several I-2 mutants substituted at phosphorylation sites (T72, S86, S120, and S121) suggested that phosphorylation of I-2 by CK2 is also involved in enhancement of association between GSK-3beta and I-2 in vivo. This study is the first demonstration that GSK-3beta associates with PP1C/I-2 complex and phosphorylates I-2 at T72 in the intact cells. 相似文献
996.
EMS16 is a member of the snake venom-derived C-type lectin family of proteins (CLPs) found in the venom of Echis multisquamatus. It binds to glycoprotein Ia/IIa (integrin alpha2beta1), a major collagen receptor of platelets, acting as a potent antagonist of platelet aggregation and cell migration. Amino acid sequencing and cDNA cloning of EMS16 have revealed that it is composed of an A chain of 134 amino acid residues and a B chain of 128 residues. Crystals of EMS16 belong to space group P2(1)2(1)2(1), with unit-cell parameters a = 46.57, b = 59.93, and c = 115.74 A, and diffract to a resolution of 1.9 A. Phase determination is underway by means of molecular replacement with the structure of blood coagulation factor IX-binding protein (IX-bp) from habu snake venom (PDB code 1bj3) as the search model. 相似文献
997.
Nomoto H Yonezawa T Itoh K Ono K Yamamoto K Oohashi T Shiraga F Ohtsuki H Ninomiya Y 《Journal of biochemistry》2003,134(2):231-238
A novel oligodendrocyte (OL)-specific cDNA was isolated from brain capillary endothelial cells and characterized. The cDNA encodes a protein of 1099 amino acids that contains a signal peptide and a transmembrane domain. The protein was expressed in mature OLs in vivo and in vitro cell cultures and was thus designated as mature OL transmembrane protein (MOLT). RT-PCR analysis showed that MOLT mRNA was expressed in brain, lung, pancreas, and testis. A polyclonal antibody raised against a part of the mouse MOLT reacted specifically with multipolar OLs possessing radially oriented processes that penetrated into the gray matter. More cells were detected in the white matter, and these had longitudinally oriented processes. In a rat OL lineage culture system, oligodendrocyte precursor cells did not initially produce MOLT mRNA and protein, but when they begun to differentiate into mature OLs, they started expressing MOLT. Consequently, MOLT may function as OLs become mature and may serve as a cell-surface marker for OL differentiation. 相似文献
998.
The microtubule-associated protein tau is a highly soluble protein that shows hardly any tendency to assemble under physiological conditions. In the brains of Alzheimer's disease (AD) patients, however, tau dissociates from the axonal microtubule and abnormally aggregates to form paired helical filaments (PHFs). One of the priorities in Alzheimer research is to clarify the mechanism of PHF formation. In recent years, several factors regulating tau assembly have come to light, yet some important questions remain to be answered. In this work, the His-tagged gene constructs of the four-repeat microtubule binding domain (4RMBD) in tau protein and its three mutants, 4RMBD S305N, N279K, and P301L, were expressed in E. coli and purified. Gel filtration chromatography and dynamic light scattering measurement yielded a Stokes radius of 3.1 nm, indicating that the His-tagged 4RMBD normally exists in buffer solution in a dimer state, which is formed by non-covalent intermolecular interactions. This non-covalent dimer can further polymerize to form filaments in the presence of polyanions such as heparin. The kinetics of the in vitro aggregation was monitored by thioflavine S dye fluorescence and CD measurements. The aggregation of 4RMBD was suggested to be a nucleation-dependent process, where the non-covalent dimer acts as an effective structural unit. The aggregation rate was strongly affected by the point mutation. Among the 4RMBD mutants, the rate of S305N was exceptionally fast, whereas N279K was the slowest, even slower than the wild-type. The aggregations were optimal in a weakly reducing environment for all the mutants and the wild type. However, the aggregations were affected differently by buffer pH, depending on the 4RMBD mutation. 相似文献
999.
Kantake N Sugiyama T Kolodner RD Kowalczykowski SC 《The Journal of biological chemistry》2003,278(26):23410-23417
Replication protein-A (RPA) is involved in many processes of DNA metabolism, including DNA replication, repair, and recombination. Cells carrying a mutation in the largest subunit of RPA (rfa1-t11: K45E) have defects in meiotic recombination, mating-type switching, and survival after DNA damage caused by UV and methyl methanesulfonate, as well as increased genome instability; however, this mutant has no significant defect in DNA replication. We purified the RPA heterotrimer containing the rfa1-t11 substitution (RPA(rfa1-t11)). This mutant RPA binds single-stranded DNA (ssDNA) with the same site size, and the RPA(rfa1-t11).ssDNA complex shows a similar sensitivity to disruption by salt as the wild-type RPA.ssDNA complex. RPA(rfa1-t11) stimulates DNA strand exchange, provided that the Rad51 protein.ssDNA nucleoprotein complex is assembled prior to introduction of the mutant RPA. However, RPA(rfa1-t11) is displaced from ssDNA by Rad51 protein more slowly than wild-type RPA and, as a consequence, Rad51 protein-mediated DNA strand exchange is inhibited when the ssDNA is in a complex with RPA(rfa1-t11). Rad52 protein can stimulate displacement of RPA(rfa1-t11) from ssDNA by Rad51 protein, but the rate of displacement remains slow compared with wild-type RPA. These in vitro results suggest that, in vivo, RPA is bound to ssDNA prior to Rad51 protein and that RPA displacement by Rad51 protein is a critical step in homologous recombination, which is impaired in the rfa1-t11 mutation. 相似文献
1000.
Maejima H Kinoshita E Seyama I Yamaoka K 《The Journal of biological chemistry》2003,278(11):9464-9471
Grayanotoxin (GTX) exerts selective effects on voltage-dependent sodium channels by eliminating fast sodium inactivation and causing a hyperpolarizing shift in voltage dependence of channel activation. In this study, we adopted a newly developed protocol that provides independent estimates of the binding and unbinding rate constants of GTX (k(on) and k(off)) to GTX sites on the sodium channel protein, important in the molecular analysis of channel modification. Novel GTX sites were determined in D2S6 (Asn-784) and D3S6 (Ser-1276) by means of site-directed mutagenesis; the results suggested that the GTX receptor consists of the S6 transmembrane segments of four homologous domains facing the ion-conducting pore. We systematically introduced at two sites in D4S6 (Na(v)1.4-Phe-1579 and Na(v)1.4-Tyr-1586) amino acid substituents with residues containing hydrophobic, aromatic, charged, or polar groups. Generally, substitutions at Phe-1579 increased both k(on) and k(off), resulting in no prominent change in dissociation constant (K(d)). It seems that the smaller the molecular size of the residue at Na(v)1.4-Phe-1579, the larger the rates of k(on) and k(off), indicating that this site acts as a gate regulating access of toxin molecules to a receptor site. Substitutions at Tyr-1586 selectively increased k(off) but had virtually no effect on k(on), thus causing a drastic increase in K(d). At position Tyr-1586, a hydrophobic or aromatic amino acid side chain was required to maintain normal sensitivity to GTX. These results suggest that the residue at position Tyr-1586 has a more critical role in mediating GTX binding than the one at position Phe-1579. Here, we propose that the affinity of GTX to Na(v)1.4 sodium channels might be regulated by two residues (Phe and Tyr) at positions Phe-1579 and Tyr-1586, which, respectively, control access and binding of GTX to its receptor. 相似文献