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81.
Human HB4C5 hybridoma cells produce a lung cancer-specific IgM human monoclonal antibody (mAb). HB4C5 human mAb cross-reacts
with Candida cytochrome c (Cyt c) and carboxypeptidase (Cpase). Concanavalin A (ConA)-resistant variants of HB4C5 cells loss
the original light chain followed by expression of various new light chains at a high incidence (light chain shifting) (Tachibana
et al., 1996). HTD8 cells, one of the ConA-resistant variant subclones of HB4C5 cells, undergo the active light chain shifting
and produce various sublines, each of which stably secretes new mAb consisting of a new light chain and a HB4C5 heavy chain.
The new mAb exhibits altered antigen binding ability from that of the original antibody. We could expect that HTD8 cells can
be used as ‘a light chain stem cell line’ to improve antigen binding ability and specificity of established human mAbs. A
BD9D12 IgG human mAb recognizes lung cancer cells and cross-reacts with cytokeratin 8. Introduction of the heavy chain gene
of BD9D12 mAb into HTD8 cells resulted in establishment of various sublines which secreted various kinds of hybrid antibodies
consisting of different light chains derived from HTD8 subclones which underwent light chain shifting and a common IgG heavy
chain derived from BD9D12. These hybrid antibodies exhibited different or improved reactivities to Cyt, Cpase, cytokeratin
8 and various cancer cells from those of parental mAb, demonstrating that light chain shifting can be applied to improve the
affinity and specificity of human mAb.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
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TNF-alpha is a key molecule in obesity-related metabolic disturbances. This study was designed to determine whether N-acetylcysteine (NAC), an antioxidant, prevents the activation of nuclear factor-kappaB (NF-kappaB) by exogenously administered TNF-alpha in adipocytes, and whether such change affects the production of adipocytokines. The treatment of well-differentiated 3T3-L1 cells with 20 mM of NAC significantly increased the reduced glutathione concentration up to 150% of control. The treatment with 10 ng/ml of TNF-alpha decreased antioxidant enzyme levels such as CuZn-superoxide dismutase (SOD), MnSOD and catalase, and activated NF-kappaB in 3T3-L1 adipocytes. The activation of NF-kappaB was significantly prevented by the pretreatment with 20 mM of NAC. TNF-alpha (1-10 ng/ml) dose-dependently increased interleukin (IL)-6 and plasminogen activator inhibitor-1 (PAI-1) secretion from 3T3-L1 adipocytes, while decreased adiponectin secretion. NAC (5-20 mM) attenuated the TNF-alpha-induced changes in these adipocytokine secretions in a dose-dependent manner. The effect of TNF-alpha and NAC on the adipocytokine productions was exerted at the m-RNA level, judging from results of the real time RT-PCR analysis. The present study revealed that NAC inhibited the TNF-alpha-mediated activation of NF-kappaB and improved the adverse changes in the levels of IL-6, PAI-1 and adiponectin in 3T3-L1 adipocytes. NAC may have the potential to improve the obesity-related abnormal adipocytokine metabolism by attenuating the TNF-alpha-induced oxidant-antioxidant imbalance in adipocytes. 相似文献
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Hiroki Murakami Sanetaka Shirahata Chikafumi Hori Hideharu Yoshii Koji Yamada Kazuki Shinohara 《Bioscience, biotechnology, and biochemistry》2013,77(8):1619-1624
Breaking activity of catecholamines and their structural analogues on λ DNA were investigated by agarose slab gel electrophoresis. Since λ DNA has a homogenous molecular size, it is a favorable material to detect the activity of DNA breaking reagent. Among the compounds tested, those having enediol group were only active, though their activities remarkably differed owing to their side chains. The profile of the breaking reaction was studied in detail by the use of one of the catecholamines, epinephrine. 相似文献
87.
The effect of variations in sodium conductances on pacemaking in a dopaminergic retinal neuron model
Shirahata T 《Acta biologica Hungarica》2011,62(2):211-214
Dopaminergic neurons in the retina show spontaneous tetrodotoxin-sensitive pacemaking, which has been explained by a reduced Hodgkin-Huxley-type computer model. The present study used this model to investigate the effect of variations in transient and persistent sodium conductance values on pacemaking, under variable leakage conductance levels. This study indicated that transient sodium conductance plays an indispensable role in pacemaking, which occurs under conditions in which only a persistent sodium conductance is considerably reduced, thus contributing to a detailed understanding of the relationship between sodium conductance and pacemaking. 相似文献
88.
DNA condensation, precipitation, and aggregation are related phenomena involving DNA-DNA interactions in the presence of multivalent cations, and studied for their potential implications in DNA packaging in the cell. Recent studies have shown that the condensation/aggregation is a prerequisite for the cellular uptake of DNA for gene therapy applications. To elucidate the ionic and structural factors involved in DNA aggregation, we studied the precipitation and resolubilization of high molecular weight and sonicated calf thymus DNA, two therapeutic oligonucleotides, and poly(dA).2Poly(dT) triplex DNA in the presence of the tetravalent polyamine spermine using a centrifugation assay, Tm measurements, and CD spectroscopy. The ability of spermine to provoke DNA precipitation was in the following order: triplex DNA > duplex DNA > single-stranded DNA. In contrast, their resolubilization at high polyamine concentrations followed a reverse order. The effective concentration of spermine to precipitate DNA increased with Na+ in the medium. Tm data indicated the DNA stabilizing effect of spermine even in the resolubilized state. CD spectroscopy revealed a series of sequential conformational alterations of duplex and triplex DNA, with the duplex form regaining the B-DNA conformation at high concentrations (approximately 200 mM) of spermine. The triplex DNA, however, remained in a Psi-DNA conformation in the resolubilized state. Chemical structural specificity effects were exerted by spermidine and spermine analogues in precipitating and resolubilizing sonicated calf thymus DNA, with N4-methyl substitution of spermidine and a heptamethylene separation of the imino groups of spermine having the maximal difference in the precipitating ability of the analogues compared to spermidine and spermine, respectively. Therapeutically important bis(ethyl) substitution reduced the precipitating ability of the analogues compared to spermine. The effect of the cationicity of polyamines was evident with the pentamines being much more efficacious than the tetramines and triamines. These results provide new insights into the mechanism of DNA precipitation by polyamines, and suggest the importance of polyamine structure in developing gene delivery vehicles for therapeutic applications. 相似文献
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The Fukushima Daiichi Nuclear Power Plant accident on March 11, 2011 attracted people’s attention, with anxiety over possible radiation hazards. Immediate and long-term concerns are around protection from external and internal exposure by the liberated radionuclides. In particular, residents living in the affected regions are most concerned about ingesting contaminated foodstuffs, including drinking water. Efficient removal of radionuclides from rainwater and drinking water has been reported using several pot-type filtration devices. A currently used flow-type test apparatus is expected to simultaneously provide radionuclide elimination prior to ingestion and protection from internal exposure by accidental ingestion of radionuclides through the use of a micro-carbon carboxymethyl cartridge unit and an electrochemically reduced water production unit, respectively. However, the removability of radionuclides from contaminated tap water has not been tested to date. Thus, the current research was undertaken to assess the capability of the apparatus to remove radionuclides from artificially contaminated tap water. The results presented here demonstrate that the apparatus can reduce radioactivity levels to below the detection limit in applied tap water containing either 300 Bq/kg of 137Cs or 150 Bq/kg of 125I. The apparatus had a removal efficiency of over 90% for all concentration ranges of radio–cesium and –iodine tested. The results showing efficient radionuclide removability, together with previous studies on molecular hydrogen and platinum nanoparticles as reactive oxygen species scavengers, strongly suggest that the test apparatus has the potential to offer maximum safety against radionuclide-contaminated foodstuffs, including drinking water. 相似文献