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71.
Elevated cortical extracellular fluid glutamate in transgenic mice expressing human mutant (G93A) Cu/Zn superoxide dismutase 总被引:6,自引:0,他引:6
Alexander GM Deitch JS Seeburger JL Del Valle L Heiman-Patterson TD 《Journal of neurochemistry》2000,74(4):1666-1673
Transgenic mice expressing a mutated (G93A) human Cu/Zn superoxide dismutase (SOD1) develop motor neuron pathology and clinical symptoms similar to those seen in patients with amyotrophic lateral sclerosis. Loss of motor neurons is most prominent in lumbar, followed by cervical cord and then brainstem. No significant cell death has been reported in motor cortex. The integrity of the cortical glutamate reuptake systems was evaluated using intracerebral microdialysis and western immunoblot assays for the glutamate transporters GLT-1, GLAST, and EAAC1. The basal extracellular fluid levels of aspartate, glutamate, glutamine, 3,4-dihydroxyphenylacetic acid, and 5-hydroxyindole-3-acetic acid were evaluated by HPLC. The extraction fraction of L-3H]glutamate, corrected with [14C]mannitol, was also evaluated. GLT-1, EAAC1, and GLAST protein levels were determined by semiquantitative chemiluminescence immunoblot of proteins from membrane-enriched fractions. The relative optical density of film was translated into relative protein level by comparison with a standard control mouse. The SOD1 mutant mice demonstrated a significant (p < 0.05) increase in basal levels of extracellular aspartate and glutamate. In addition, when the glutamate extraction fraction was challenged with exogenous unlabeled glutamate (500 microM) by reversed microdialysis, the glutamate extraction fraction in the mutant SOD1 mice was decreased significantly from control levels. The SOD1 mutant mice demonstrated no difference in the cortical protein levels of the glutamate transporter subtypes. This study demonstrates that in areas of no visible pathology and no loss of glutamate transporter proteins, SOD1 mutant mice have elevated extracellular fluid aspartate and glutamate levels and a decreased capacity to clear glutamate from the extracellular space. 相似文献
72.
73.
中华姬鼠与大林姬鼠的同工酶差异 总被引:1,自引:0,他引:1
中华姬鼠(Apodemusdraco)和大林姬鼠(Apodemuspeninsulae)是形态学上十分相似的两种鼠类。为了对两种姬鼠的分类提供生物化学方面的依据,采用聚丙烯酰胺凝胶等电聚焦电泳方法比较和分析了两种姬鼠的LDH同工酶、EST同工酶和SOD同工酶的差异。结果表明,两种姬鼠的LDH同工酶酶谱基本相似,而EST同工酶和SOD同工酶酶谱则存在明显的种间差异。根据EST同工酶A2带的有无和SOD同工酶主带等电点的差别,能将两种姬鼠很容易区分开来。 相似文献
74.
75.
参麦注射液抗心肌缺氧-再给氧损伤实验研究 总被引:3,自引:0,他引:3
采用Langendorf离体心脏灌注模型,对大鼠心肌缺氧—再给氧损伤中抗自由基酶SOD和GSH-Px,过氧化产物MDA、心肌酶CPK和心肌细胞超微结构进行了观察、同时探讨了参麦注射液的保护作用机理。结果表明:(1)心肌缺氧灌注40min,富氧再灌5min,与正常对照组比较,心肌细胞超微结构损伤严重,线粒体数目减少,大部分空泡变性,嵴消失,糖原颗粒减少,心肌收缩结构受到严重破坏。同时CPK活性明显升高,SOD及GSH-Px活性明显降低,MDA含量明显升高(P<0.01)。(2)预先给不同剂量参麦注射液进行灌注,与模型组比较,心肌超微结构损伤明显减轻,线粒体数目较多,嵴密集,未见肿胀变形,糖原颗粒丰富,心肌收缩结构基本正常。CPK活性明显降低,心肌SOD及GSH-Px活性明显增高,心肌MDA含量明显降低(P<0.01)。且参麦大剂量组疗效优于复方丹参液(P<0.05)。我们推测其保护作用机理可能是稳定心肌细胞膜,保护心肌线粒体,增加能量供应,提高抗自由基酶活性,从而减轻氧自由基对心肌的损害 相似文献
76.
A cDNA clone for the cytosolic Cu/Zn superoxide dismutase (Cu/Zn SOD) from Chinese cabbage (Brassica campestris ssp.pekinensis) was isolated and its DNA sequence was determined. The cDNA clone contains a complete coding sequence which encodes a protein
of 152 amino acids and a 3-untranslated region including a poly A signal. The deduced amino acid sequence shows that it is
highly homologous to the Cu/Zn SODs from other plants (60–90%). The lack of a putative chloroplast targeting transit peptide
indicates that the clone represents a cytosolic form of Cu/Zn SOD. Genomic Southern hybridization suggests that cytosolic
Cu/Zn SOD genes are present in 1 or 2 copies per genome. 相似文献
77.
The enzymes involved in the protection of plant metabolism in presence of azo dye was characterized by studying activities of the role of antioxidant enzymes in the hairy roots (HRs) of Physalis minima L. during degradation of an azo dye, Reactive Black 8 (RB8). When the HRs were exposed to RB8 (30 mg L?1), a nine fold increase in SOD activity was observed after 24 h, while 22 and 50 fold increase in activity was observed for POX and APX respectively after 72 h, whereas there was no significant change in activity of CAT. The activation of different antioxidant enzymes at different time intervals under dye stress suggests the synchronized functioning of antioxidant machinery to protect the HRs from oxidative damage. FTIR analysis confirmed the degradation of dye and the non-toxic nature of metabolites formed after dye degradation was confirmed by phytotoxicity study. 相似文献
78.
Seiji Miyake Saori Kobayashi Kazuo Tsubota Yoko Ozawa 《Biochemical and biophysical research communications》2014
The mechanism by which lutein, a carotenoid, acts as an antioxidant in retinal cells is still not fully understood. Here, lutein treatment of a neuronal cell line (PC12D) immediately resulted in reduced intracellular ROS levels, implying that it has a direct role in ROS scavenging. Significantly, lutein treatment also induced phase II antioxidative enzyme expression, probably via a nuclear factor-like 2 (Nrf2) independent pathway. This latter mechanism could explain why lutein acts diversely to protect against oxidative/cytotoxic stress, and why it is physiologically involved in the human neural tissue, such as the retina. 相似文献
79.
Xi Cheng Xingjian HuangSiyu Liu Mi TangWanfeng Hu Siyi Pan 《Biochemical and biophysical research communications》2014
Polyphenol oxidases (PPOs) catalyzing the oxygen dependent oxidation of phenols to quinones are ubiquitously distributed in plants and are assumed to be involved in plant defense against pests and pathogens. A protein with high PPO activity was identified in Satsuma mandarine, extracted with Tris–HCl buffer, purified by salt precipitation and column chromatography, and characterized by mass spectrometry as germin-like protein (GLP), which belongs to pathogenesis related protein (PR) family. In the present study, the structure and enzymatic properties of GLP were characterized using spectroscopy methods. Based on native PAGE analysis, the molecular weight of GLP was estimated to be 108 kDa and GLP was identified as a pentamer containing five subunits of 22 kDa. The optimum pH and temperature for PPO catalyzing activity of GLP was 6.5 and 65 °C, respectively. Kinetic constants were 0.0365 M and 0.0196 M with the substrates catechol and pyrogallol, respectively. The structural characterization of GLP provided better insights into the regions responsible for its PPO activity. 相似文献
80.