全文获取类型
收费全文 | 1007篇 |
免费 | 65篇 |
国内免费 | 17篇 |
出版年
2023年 | 16篇 |
2022年 | 23篇 |
2021年 | 16篇 |
2020年 | 24篇 |
2019年 | 30篇 |
2018年 | 36篇 |
2017年 | 22篇 |
2016年 | 14篇 |
2015年 | 22篇 |
2014年 | 34篇 |
2013年 | 51篇 |
2012年 | 21篇 |
2011年 | 25篇 |
2010年 | 25篇 |
2009年 | 37篇 |
2008年 | 55篇 |
2007年 | 42篇 |
2006年 | 35篇 |
2005年 | 40篇 |
2004年 | 43篇 |
2003年 | 26篇 |
2002年 | 33篇 |
2001年 | 19篇 |
2000年 | 18篇 |
1999年 | 22篇 |
1998年 | 19篇 |
1997年 | 24篇 |
1996年 | 17篇 |
1995年 | 35篇 |
1994年 | 23篇 |
1993年 | 12篇 |
1992年 | 16篇 |
1991年 | 16篇 |
1990年 | 14篇 |
1989年 | 22篇 |
1988年 | 13篇 |
1987年 | 7篇 |
1986年 | 6篇 |
1985年 | 10篇 |
1984年 | 24篇 |
1983年 | 11篇 |
1982年 | 21篇 |
1981年 | 12篇 |
1980年 | 13篇 |
1979年 | 8篇 |
1977年 | 7篇 |
1976年 | 6篇 |
1975年 | 5篇 |
1974年 | 4篇 |
1973年 | 4篇 |
排序方式: 共有1089条查询结果,搜索用时 15 毫秒
31.
Modulation of Ion Gradients and Glutamate Release in Cultured Cerebellar Granule Cells by Ouabain 总被引:8,自引:0,他引:8
Abstract: Upon addition of the cardiac glycoside ouabain to cultured cerebellar granule cells, an immediate increase in intracellular free sodium is evoked mediated by two pathways, a voltage-sensitive channel blocked by tetrodotoxin and a channel sensitive to flunarizine. Ouabain induces a steady plasma membrane depolarization in low Ca2+ medium; whereas in the presence of Ca2+ , a distinct discontinuity is observed always preceded by a large increase in intracellular free Ca2+ ([Ca2+ ]c ). The plateau component of the increase can be inhibited additively by the L-type Ca2+ channel antagonist nifedipine, the spider toxin Aga-Gl, and the NMDA receptor antagonist MK-801. Single-cell imaging reveals that the [Ca2+ ]c increase occurs asynchronously in the cell population and is not dependent on a critical level of extracellular glutamate or synaptic transmission between the cells. A prolonged release of glutamate is also observed that is predominantly Ca2+ dependent for the first 6–10 min after the evoked increase in [Ca2+ ]c . This release is four times as large as that observed with 50 m M KCl and is predominantly exocytotic because release was inhibited by tetanus toxin, the V-type ATPase inhibitor bafilomycin, and Aga-Gl. It is proposed, therefore, that ouabain induces a period of membrane excitability culminating in a sustained exocytosis above that observed upon permanent depolarization with KCl. 相似文献
32.
N-terminal sequence analysis of atrial granule serine proteinase purified by affinity chromatography
Atrial granule serine proteinase is considered the leading candidate endoproteolytic processing enzyme of pro-atrial natriuretic factor. Its cleavage specificity is directed toward a monobasic amino acid processing site, and as such, the atrial enzyme is distinguished from the family of prohormone convertases which act at dibasic amino acid processing sites. To delineate the molecular mechanisms which distinguish monobasic from dibasic amino acid-directed processing enzymes, pure atrial enzyme is needed for sequence determination leading to molecular cloning, and for preparation of antisera. An affinity chromatography purification scheme seemed a logical modification of our established procedures to yield suitable amounts of enzyme for further studies. Surprisingly, pseudo-peptide bond inhibitors of the atrial enzyme [Damodaran and Harris (1995),J. Protein Chem., this issue] formed ineffective affinity ligands, even though these compounds contain essential residues on either side of what would be the scissile bond in a peptide substrate. On the other hand, tripeptide aldehydes (based on the substrate recognition sequence of the atrial enzyme) linked to Sepharose formed effective affinity matrices, permitting purification of the enzyme in a single step from a subcellular fraction enriched for atrial granules and lysosomes. Hence, the enzyme was purified 2000-fold in 90% overall yield, and subjected to N-terminal sequence analysis through 26 residues. The sequence determined, XXPEAAGLPG[R, L]GNPVP[F, G]R[Q, I]XY[G, E]XR(N, A]V, indicates that the atrial enzyme is unique, showing little sequence homology to other proteins in the database.Abbreviations AGSP
atrial granule serine proteinase
- ANF
atrial natriuretic factor
- BSA
bovine serum albumin
-
Bz
benzoyl
- EACA
6()-aminocaproic acid
- HEPES
N-2-hydroxyethylpiperazine-N'-propanesulfonic acid
- HPLC
high-performance liquid chromatography
- PEG
polyethylene glycol-3350
- SDS-PAGE
sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Single-letter abbreviations are used to denote amino acids 相似文献
33.
Pseudo-peptide bond inhibitors (-bond inhibitors) and peptide-aldehyde inhibitors of atrial granule serine proteinase, the candidate processing enzyme of pro-atrial natrieuretic factor, are prepared in high yield and purity by novel synthetic routes. The -bond compounds retain essential residues for enzyme binding, but place the enzyme inhibition site in the midst of the peptide sequence. Thus, Bz-APR--LR and Bz-APR--SLRR can be considered readthrough inhibitors of atrial granule serine proteinase. The most potent -peptide, Bz-APR--SLRR (IC50=250 M), is about fivefold less potent than the best peptide-aldehyde inhibitor (EACA-APR-CHO), and both the -bond and peptide-aldehyde compounds are competitive, reversible inhibitors of the enzyme. The -bond peptides containing two C-terminal Arg residues are three-to tenfold more potent than the analogous compounds containing only one C-terminal Arg residue, confirming the importance of both Arg residues in the enzyme processing recognition site. As expected, because of their moderate potencies, the -peptides are not useful affinity ligands for purification of atrial granule serine proteinase, but both peptide aldehydes are effective affinity ligands [Damodaran and Harris (1995),J. Protein Chem., this issue].Abbreviations AGSP
atrial granule serine proteinase
- ANF
atrial natriuretic factor
- Bz
benzoyl
- DIEA
diisopropylethylamine
- DIPCDI
diisopropylcarbodiimide
- DMF
dimethylformamide
- DMSO
dimethylsulfoxide
- EACA
6(e)-aminocaproic acid
- EtOAc
ethyl acetate
- HEPES
N-2-hydroxyethylpiperazine-N-propanesulfonic acid
- HOBt
N-hydroxybenzotriazole
- HPLC
high-performance liquid chrornatography
- NMR
nuclear magnetic resonance
- PEG
polyethylene glycol-3350
- PyBOP
benzotriazole-1-yl-oxy-trispyrrolidino-phosphonium-hexafluorophospate
- TEA
triethylamine
- TFA
trifluoroacetic acid
- THF
tetrahydrofuran
- TLC
thin-layer chromatography
- UV
ultraviolet
-
pseudo-peptide bond -CH2-NH-. Single-letter abbreviations are used to denote amino acids 相似文献
34.
Ethanol Promotes Apoptosis in Cerebellar Granule Cells by Inhibiting the Trophic Effect of NMDA 总被引:8,自引:3,他引:5
Abstract: When primary cultures of cerebellar granule neurons are grown in a physiological concentration of KCl (5 m M ) they undergo apoptosis, which can be prevented by growing the cells in the presence of N -methyl- d -aspartate (NMDA). We now show that ethanol inhibits this trophic effect of NMDA, i.e., promotes apoptosis, and also inhibits the NMDA-induced increase in intracellular Ca2+ concentration in cells grown in 5 m M KCl. Both effects of ethanol show a similar concentration dependence and are reversed by a high concentration of glycine, the co-agonist at the NMDA receptor. The data suggest that the effect of ethanol on apoptosis is mediated, at least in part, by inhibition of NMDA receptor function. This effect of ethanol to increase apoptosis could contribute to the previously described in vivo sensitivity of the developing cerebellum to ethanol-induced damage. 相似文献
35.
Nurit Nardi Galya Avidan Dvorah Daily Rina Zilkha-Falb Ari Barzilai 《Journal of neurochemistry》1997,68(2):750-759
Abstract: We analyzed biochemically and temporally the molecular events that occur in the programmed cell death of mouse cerebellar granule neurons deprived of high potassium levels. An hour after switching the neurons to a low extracellular K+ concentration ([K+]o), a significant part of the genomic DNA was already cleaved to high-molecular-weight fragments. This phenomenon was intensified with the progression of the death process. Addition of cycloheximide to the neurons 4 h after high [K+]o deprivation resulted in no cell loss and complete recovery of the damaged DNA. DNA margination and nuclear fragmentation as assessed by 4,6-diaminodiphenyl-2-phenylindole staining were observable in a few cells beginning ~4 h after the removal of high [K+]o and developed to nuclear condensation 4 h later. Six hours after high [K+]o deprivation, the DNA was fragmented into oligonucleosome-sized fragments. Within 6 h after removal of the extracellular K+, 50% of the neurons were committed to die and lost their ability to be rescued by readministration of 25 mM [K+]o. Similar to high [K+]o deprivation, inhibition of RNA or protein synthesis failed to halt neuronal degeneration of a similar percentage of cells 6 h after the onset of the death process. Mitochondrial function steadily decreased after [K+]o removal. An ~40% decrease in RNA and protein synthesis was detected by 6 h of [K+]o removal during the period of cell death commitment; rates continued to decline gradually thereafter. The temporal characteristics of the DNA damage and recovery, DNA cleavage to oligonucleosome-sized fragments, and the reduction in mitochondrial activity—events that occurred within the critical time—may indicate that these processes have an important part in the mechanism that committed the neurons to die. 相似文献
36.
N-Acetylaspartylglutamate Stimulates Metabotropic Glutamate Receptor 3 to Regulate Expression of the GABAA α6 Subunit in Cerebellar Granule Cells 总被引:1,自引:1,他引:0
Subroto Ghose Barbara Wroblewska Lorenzo Corsi †Dennis R. Grayson ‡Angel L. De Blas Stefano Vicini Joseph H. Neale 《Journal of neurochemistry》1997,69(6):2326-2335
Abstract: We have shown that the vertebrate neuropeptide N-acetylaspartylglutamate (NAAG) meets the criteria for a neurotransmitter, including function as a selective metabotropic glutamate receptor (mGluR) 3 agonist. Short-term treatment of cerebellar granule cells with NAAG (30 µM) results in the transient increase in content of GABAAα6 subunit mRNA. Using quantitative PCR, this increase was determined to be up to 170% of control values. Similar effects are seen following treatment with trans-1-aminocyclopentane-1,3-dicarboxylate and glutamate and are blocked by the mGluR antagonists (2S,3S,4S)-2-methyl-2-(carboxycyclopropyl)glycine and (2S)-α-ethylglutamic acid. The effect is pertussis toxin-sensitive. The increase in α6 subunit mRNA level can be simulated by activation of other receptors negatively linked to adenylate cyclase activity, such as adenosine A1, α2-adrenergic, muscarinic, and GABAB receptors. Forskolin stimulation of cyclic AMP (cAMP) levels abolished the effect of NAAG. The change in α6 levels induced by 30 µM NAAG can be inhibited in a dose-dependent manner by simultaneous application of increasing doses of the β-adrenergic receptor agonist isoproterenol. The increase in α6 mRNA content is followed by a fourfold increase in α6 protein level 6 h posttreatment. Under voltage-clamped conditions, NAAG-treated granule cells demonstrate an increase in the furosemide-induced inhibition of GABA-gated currents in a concentration-dependent manner, indicating an increase in functional α6-containing GABAA receptors. These data support the hypothesis that NAAG, acting through mGluR3, regulates expression of the GABAAα6 subunit via a cAMP-mediated pathway and that cAMP-coupled receptors for other neurotransmitters may similarly influence GABAA receptor subunit composition. 相似文献
37.
A. Christine Engblom Michael J. Courtney Jyrki P. Kukkonen Karl E. O. Åkerman 《Journal of neurochemistry》1997,69(5):2162-2168
Abstract: The effect of ethanol on the intracellular Ca2+ concentration response to NMDA in rat cerebellar granule cells grown in low or high KCI concentrations has been studied using image analysis. The cells grown in low KCI displayed high sensitivity for glycine. The subtype-selective antagonist ifenprodil inhibited the response with high (in the low micromolar range) and low (in the high micromolar range) potency. Ethanol affected the high-potency component in these cultures. In cells grown in high KCI the glycine sensitivity was lower, and a low potency for ifenprodil (high micromolar) dominated. These cells were not significantly sensitive to ethanol. The results indicate that the component displaying potency for ifenprodil in the low micromolar range with properties of the NR2B subunit is the target for ethanol action on the NMDA receptor. 相似文献
38.
本文报道大鼠水盐负荷时,其心房特殊颗粒(ASG)的数目及钙含量发生相关变化。给大鼠禁水5d或饮2%NaCl液4d造成水盐负荷,用电镜形态计量法计数ASG,以反映心房钠尿肽(ANP)的分泌水平,电镜X射线显微分析法测定ASG中钙、硫含量及肌质同终末池(TSR)中钙含量。结果显示,禁水引起ANP分泌减少时,ASG的数密度增加(6.02±2.30增至9.97±3.21个/μm3),同时伴有钙含量增加(64±16增至92±18mmol/kg);饮盐水引起ANP分泌增加时,ASG的数密度减少(6.02±2.30减至2.96±1.62个/μm3),巨伴有钙含量减少(64±16减至38±22mmol/kg),但水盐负荷对TSR中钙浓度无任何影响。据此推论ASG作为细胞内钙库,借某种机制调控其中钙的释放而参与ANP的刺激分泌耦联过程。 相似文献
39.
A selective uptake mechanism for some nucleosides and related substances was found in retinae of light adapted rabbits and fish. After the intravitreal injection in vivo of [3H]adenosine, [3H]inosine, [3H]guanosine and certain related compounds, the distribution of radioactivity was studied by autoradiography. Retinae were also incubated in [3H]adenosine and [3H]inosine and then were similarly processed.In rabbits, the accumulation of radioactivity from [3H]adenosine and [3H]guanosine was predominantly into glial cells, but also into neurons. [3H]Inosine labelled glia almost exclusively. However, the adenosine analog, [3H]methylphenylethyl-adenosine, resulted in well-defined neuronal labelling in this species. In fish, a few photoreceptor cell bodies exhibited strong radioactivity with the nucleosides, presumably representing incorporation into nucleic acids of replicating cells. Labelling was also seen in horizontal cells, amacrine cells and ganglion cells after the injection of either [3H]adenosine, [3H]guanosine or [3H]inosine.To some extent, the selective accumulation of radioactivity is likely to be due to cell replication, but in most neurons, other factors must be responsible. Judging from what is known about the actions of adenosine in central nervous tissue, signal transmission in the retina could be such a factor. 相似文献
40.
(1) Redox titrations of cytochrome b-561 have been performed with the purified cytochrome and with intact and detergent-solubilized chromaffin-granule membranes. (2) The midpoint redox potential of the cytochrome is 100–130 mV; this depends upon the composition of the buffer, but is independent of pH in the range 5.5–7.5; partial proteolysis of the cytochrome raises the midpoint potential to 160 mV. (3) The Nernst plots of titration data have slopes of 75–115 mV, and are in some cases sigmoid in shape. This may be explained by negative cooperativity during redox transitions in oligomeric cytochrome b-561. (4) Measurements of the haem and cytochrome content of chromaffin granule membrane suggest a haem content of 1 mol/mol protein. (5) Chemical crosslinking of cytochrome b-561 suggests that it may exist as an oligomer of 4–6 polypeptide chains within the chromaffin granule membrane. Aggregation of purified cytochrome b-561 was shown by gel filtration studies and by immunological methods in SDS-polyacrylamide gels. Studies of the molecular weight of the aggregates suggest that the monomer has a molecular weight close to 22 000, but migrates anomalously slowly during electrophoresis. 相似文献