全文获取类型
收费全文 | 29331篇 |
免费 | 4165篇 |
国内免费 | 15657篇 |
专业分类
49153篇 |
出版年
2024年 | 360篇 |
2023年 | 1062篇 |
2022年 | 1700篇 |
2021年 | 1831篇 |
2020年 | 1788篇 |
2019年 | 2012篇 |
2018年 | 1418篇 |
2017年 | 1402篇 |
2016年 | 1446篇 |
2015年 | 1981篇 |
2014年 | 2717篇 |
2013年 | 2358篇 |
2012年 | 3189篇 |
2011年 | 3118篇 |
2010年 | 2478篇 |
2009年 | 2511篇 |
2008年 | 2727篇 |
2007年 | 2459篇 |
2006年 | 2255篇 |
2005年 | 1861篇 |
2004年 | 1524篇 |
2003年 | 1232篇 |
2002年 | 1111篇 |
2001年 | 953篇 |
2000年 | 897篇 |
1999年 | 597篇 |
1998年 | 305篇 |
1997年 | 186篇 |
1996年 | 172篇 |
1995年 | 132篇 |
1994年 | 127篇 |
1993年 | 119篇 |
1992年 | 116篇 |
1991年 | 79篇 |
1990年 | 96篇 |
1989年 | 77篇 |
1988年 | 76篇 |
1987年 | 61篇 |
1986年 | 66篇 |
1985年 | 63篇 |
1984年 | 55篇 |
1983年 | 41篇 |
1982年 | 61篇 |
1981年 | 32篇 |
1980年 | 28篇 |
1977年 | 13篇 |
1964年 | 13篇 |
1957年 | 14篇 |
1953年 | 14篇 |
1950年 | 20篇 |
排序方式: 共有10000条查询结果,搜索用时 0 毫秒
81.
82.
83.
An internal deletion in the cytoplasmic tail reverses the apical localization of human NGF receptor in transfected MDCK cells 总被引:16,自引:9,他引:7
A Le Bivic Y Sambuy A Patzak N Patil M Chao E Rodriguez-Boulan 《The Journal of cell biology》1991,115(3):607-618
A cDNA encoding the full-length 75-kD human nerve growth factor receptor was transfected into MDCK cells and its product was found to be expressed predominantly (80%) on the apical membrane, as a result of vectorial targeting from an intracellular site. Apical hNGFR bound NGF with low affinity and internalized it inefficiently (6% of surface bound NGF per hour). Several mutant hNGFRs were analyzed, after transfection in MDCK cells, for polarized surface expression, ligand binding, and endocytosis. Deletionof juxta-membrane attachment sites for a cluster of O-linked sugars did not alter apical localization. A mutant receptor lacking the entire cytoplasmic tail (except for the five proximal amino acids) was also expressed on the apical membrane, suggesting that information for apical sorting was contained in the ectoplasmic or transmembrane domains. However, a 58 amino acid deletion in the hNGFR tail that moved a cytoplasmic tyrosine (Tyr 308) closer to the membrane into a more charged environment resulted in a basolateral distribution of the mutant receptor and reversed vectorial (basolateral) targeting. The basolateral mutant receptor also internalized 125I-NGF rapidly (90% of surface bound NGF per hour), exhibited a larger intracellular fraction and displayed a considerably shortened half-life (approximately 3 h). We suggest that hNGFR with the internal cytoplasmic deletion expresses a basolateral targeting signal, related to endocytic signals, that is dominant over apical targeting information in the ecto/transmembrane domains. These results apparently contradict a current model that postulates that basolateral targeting is a default mechanism. 相似文献
84.
The regulation of tissue kallikrein activity by plasma serine proteinase inhibitors (serpins) was investigated by measuring the association rate constants of six tissue-kallikrein family members isolated from the rat submandibular gland, with rat kallikrein-binding protein (rKBP) and alpha 1-proteinase inhibitor (alpha 1-PI). Both these serpins inhibited kallikreins rK2, rK7, rK8, rK9 and rK10 with association rate constants in the 10(3)-10(4) M-1.s-1 range, whereas only 'true' tissue kallikrein rK1 was not susceptible to alpha 1-PI. This results in slow inhibition of rK1 by plasma serpins, which could explain why this kallikrein is the only member of the gene family identified so far that induces a transient decrease in blood pressure when injected in minute amounts into the circulation. 相似文献
85.
Activation of MAP kinases by calcium-dependent and calcium-independent pathways. Stimulation by thapsigargin and epidermal growth factor. 总被引:5,自引:0,他引:5
T S Chao K L Byron K M Lee M Villereal M R Rosner 《The Journal of biological chemistry》1992,267(28):19876-19883
In order to determine the effect of calcium mobilization on mitogen-activated protein (MAP) kinase activation, we have treated human foreskin fibroblasts (HSWP cells) and human epidermal carcinoma (A431) cells with thapsigargin. Intracellular free calcium was monitored by single cell image analysis using fura-2 and correlated with MAP kinase stimulation as assessed by immunoprecipitation, kinase renaturation assays and immunoblotting. Thapsigargin stimulated the 44- and 42-kDa MAP kinase isozymes in both cell types with kinetics that were slightly delayed relative to enzyme stimulated by epidermal growth factor. Removal of external calcium did not significantly affect the activation of the MAP kinases by thapsigargin, indicating that intracellular calcium mobilization is sufficient to stimulate the enzymes. However, treatment of cells with EGTA under conditions which deplete both intra- and extracellular calcium inhibited stimulation by thapsigargin but not epidermal growth factor. Stimulation of the MAP kinases by the calcium ionophore ionomycin paralleled the activation observed with thapsigargin in both calcium-containing and calcium-free conditions. These results indicate that there are at least two independent pathways for stimulation of MAP kinase: one that is dependent on intracellular calcium mobilization, and one that is mediated by the tyrosine kinase epidermal growth factor receptor and is calcium-independent. 相似文献
86.
A novel human tissue kallikrein inhibitor designated as kallistatin has been purified from plasma to apparent homogeneity by polyethylene glycol fractionation and successive chromatography on heparin-Agarose, DEAE-Sepharose, hydroxylapatite, and phenyl-Superose columns. A purification factor of 4350 was achieved with a yield of approximately 1.35 mg per liter of plasma. The purified inhibitor migrates as a single band with an apparent molecular mass of 58 kDa when analyzed on SDS-polyacrylamide gel electrophoresis under reducing conditions. It is an acidic protein with pI values ranging from 4.6 to 5.2. No immunological cross-reactivity was found by Western blot analyses between kallistatin and other serpins. Kallistatin inhibits human tissue kallikrein's activity toward kininogen and tripeptide substrates. The second-order reaction rate constant (ka) was determined to be 2.6 x 10(4) M-1 s-1 using Pro-Phe-Arg-MCA. The inhibition is accompanied by formation of an equimolar, heat- and SDS-stable complex between tissue kallikrein and kallistatin, and by generation of a small carboxyl-terminal fragment from the inhibitor due to cleavage at the reactive site by tissue kallikrein. Heparin blocks kallistatin's complex formation with tissue kallikrein and abolishes its inhibitory effect on tissue kallikrein's activity. The amino-terminal residue of kallistatin is blocked. Sequence analysis of the carboxyl-terminal fragment generated from kallistatin reveals the reactive center sequence from P1' to P15', which shares sequence similarity with, but is different from known serpins including protein C inhibitor, alpha 1-antitrypsin, and alpha 1-antichymotrypsin. The results show that kallistatin is a new member of the serpin superfamily that inhibits human tissue kallikrein. 相似文献
87.
We studied the effects of arachidonic acid (AA) on Cl secretion across primary cultures of dog tracheal epithelium. Cell sheets showed mean values for baseline short-circuit current (Isc) and transepithelial resistance of 33.8 muA/cm2 and 360 omega.cm2 (n = 44). AA (5 x 10(-5) M) added to both sides increased Isc by 27.8 +/- 5.2 muA/cm2 (mean +/- SE, n = 8), and elevated intracellular cAMP levels. In the presence of 5 x 10(-6) M of both indomethacin (INDO) and nordihydroguaiaretic acid (NDGA) (inhibitors of cyclooxygenase and lipoxygenase, respectively), AA reduced Isc by 4.4 +/- 0.6 muA/cm2 (n = 10) without changing cAMP. Both INDO and NDGA were necessary to abolish the Isc increase in response to AA. The effects of AA on Isc were unaffected by amiloride. In the presence of INDO and NDGA, isoproterenol (ISO) raised cAMP and increased Isc by 27.6 +/- 4.3 (transient) and 12.8 +/- 3.2 muA/cm2 (sustained) (n = 9). With AA present as well as INDO and NDGA, the transient and sustained responses to ISO were significantly reduced to 13.2 +/- 2.4 and 3.9 +/- 0.8 muA/cm2 (n = 10), respectively; the increase in cAMP was unaltered. AA approximately halved baseline efflux of 125I from confluent cell sheets in high K medium and reduced the isoproterenol-induced increase in efflux to 20% of control. These data are consistent with the reported inhibitory effect of AA on apical membrane chloride channels. 相似文献
88.
89.
本文报道了寄生在云杉上的中国新记录种顶裂盘菌(Lophophacidium hyperboreumLagerb.);首次发现了这个种的无性型座壳梭孢属(Apostrasseria sp.),证实了融雪前病株针叶上有表生的菌丝和小菌核;查清了它是新疆云杉林中的广布种,引致云杉雪枯病;发现了新分布区,地理分布范围在75°—94°E,37°40′—49°N;发现了新寄主,多土的西伯利亚云杉(Picea obovata Ledeb.)和引进的青海云杉(P.crassifolia Kom.)、川西云杉(P.balfourianaRehd.et Wils.)。 相似文献
90.
Deletion of cytoplasmic sequences of the nerve growth factor receptor leads to loss of high affinity ligand binding 总被引:13,自引:0,他引:13
The nerve growth factor (NGF) receptor is a glycosylated transmembrane protein present on the cell surface as both high and low affinity forms, but biological responsiveness requires interactions of NGF with the high affinity site. We have tested the effects of mutations in the intracellular domain of the receptor upon its cell surface expression and equilibrium binding of 125I-NGF. Although mutant receptors lacking the entire cytoplasmic domain are processed and expressed at the cell surface and are capable of binding to NGF, the absence of cytoplasmic sequences leads to a loss of high affinity binding and to a lack of an appropriate cross-linking pattern as assessed by N-hydroxysuccinimidyl 4-azidobenzoate photoaffinity cross-linking. These results, taken together with the highly conserved nature of these cytoplasmic sequences, implies that the interaction of the receptor with an accessory molecule is necessary to form the high affinity receptor. 相似文献