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Ertan Kucuksayan Aysegul Cort Mujgan Timur Evrim Ozdemir Suleyman Gultekin Yucel Prof. Dr. Tomris Ozben 《Journal of cellular biochemistry》2013,114(7):1685-1694
Antioxidants may prevent apoptosis of cancer cells via inhibiting reactive oxygen species (ROS). However, to date no study has been carried out to elucidate the effects of strong antioxidant N‐acetylcysteine (NAC) on Bleomycin induced apoptosis in human testicular cancer (NTERA‐2, NT2) cells. For this reason, we studied the effects of Bleomycin and NAC alone and in combination on apoptotic signaling pathways in NT2 cell line. We determined the cytotoxic effect of bleomycin on NT2 cells and measured apoptosis markers such as Caspase‐3, ‐8, ‐9 activities and Bcl‐2, Bax, Cyt‐c, Annexin V‐FTIC and PI levels in NT2 cells incubated with different agents for 24 h. Early apoptosis was determined using FACS assay. We found half of the lethal dose (LD50) of Bleomycin on NT2 cell viability as 400, 100, and 20 µg/ml after incubations for 24, 48, and 72 h, respectively. Incubation with bleomycin (LD50) and H2O2 for 24 h increased Caspase‐3, ‐8, ‐9 activities, Cyt‐c and Bax levels and decreased Bcl‐2 levels. The concurrent incubation of NT2 cells with bleomycin/H2O2 and NAC (5 mM) for 24 h abolished bleomycin/H2O2‐dependent increases in Caspase‐3, ‐8, ‐9 activities, Bax and Cyt‐c levels and bleomycin/H2O2‐dependent decrease in Bcl‐2 level. Our results indicate that bleomycin/H2O2 induce apoptosis in NT2 cells by activating mitochondrial pathway of apoptosis, while NAC diminishes bleomycin/H2O2 induced apoptosis. We conclude that NAC has antagonistic effects on Bleomycin‐induced apoptosis in NT2 cells and causes resistance to apoptosis which is not a desired effect in eliminating cancer cells. J. Cell. Biochem. 114: 1685–1694, 2013. © 2013 Wiley Periodicals, Inc. 相似文献
13.
Priscilla F. McAuliffe Kurt W. Evans Argun Akcakanat Ken Chen Xiaofeng Zheng Hao Zhao Agda Karina Eterovic Takafumi Sangai Ashley M. Holder Chandeshwar Sharma Huiqin Chen Kim-Anh Do Emily Tarco Mihai Gagea Katherine A. Naff Aysegul Sahin Asha S. Multani Dalliah M. Black Elizabeth A. Mittendorf Isabelle Bedrosian Gordon B. Mills Ana Maria Gonzalez-Angulo Funda Meric-Bernstam 《PloS one》2016,11(3)
14.
Erkan Tuncay Yusuf Olgar Aysegul Durak Sinan Degirmenci Ceylan Verda Bitirim Belma Turan 《Journal of cellular physiology》2019,234(8):13370-13386
Role of β3-AR dysregulation, as either cardio-conserving or cardio-disrupting mediator, remains unknown yet. Therefore, we examined the molecular mechanism of β3-AR activation in depressed myocardial contractility using a specific agonist CL316243 or using β3-AR overexpressed cardiomyocytes. Since it has been previously shown a possible correlation between increased cellular free Zn2+ ([Zn2+]i) and depressed cardiac contractility, we first demonstrated a relation between β3-AR activation and increased [Zn2+]i, parallel to the significant depolarization in mitochondrial membrane potential in rat ventricular cardiomyocytes. Furthermore, the increased [Zn2+]i induced a significant increase in messenger RNA (mRNA) level of β3-AR in cardiomyocytes. Either β3-AR activation or its overexpression could increase cellular reactive oxygen species (ROS) and reactive nitrogen species (RNS) levels, in line with significant changes in nitric oxide (NO)-pathway, including increases in the ratios of pNOS3/NOS3 and pGSK-3β/GSK-3β, and PKG expression level in cardiomyocytes. Although β3-AR activation induced depression in both Na+- and Ca2+-currents, the prolonged action potential (AP) seems to be associated with a marked depression in K+-currents. The β3-AR activation caused a negative inotropic effect on the mechanical activity of the heart, through affecting the cellular Ca2+-handling, including its effect on Ca2+-leakage from sarcoplasmic reticulum (SR). Our cellular level data with β3-AR agonism were supported with the data on high [Zn2+]i and β3-AR protein-level in metabolic syndrome (MetS)-rat heart. Overall, our present data can emphasize the important deleterious effect of β3-AR activation in cardiac remodeling under pathological condition, at least, through a cross-link between β3-AR activation, NO-signaling, and [Zn2+]i pathways. Moreover, it is interesting to note that the recovery in ER-stress markers with β3-AR agonism in hyperglycemic cardiomyocytes is favored. Therefore, how long and to which level the β3-AR agonism would be friend or become foe remains to be mystery, yet. 相似文献
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We have improved the expression of recombinant human granulocyte-colony-stimulating factor (G-CSF), produced by either pL or trpP expression vectors in Escherichia coli, by altering the sequence at the 5' end of the G-CSF-coding region. Initial attempts to express G-CSF resulted in neither detectable G-CSF mRNA nor protein in the trpP system, and only G-CSF mRNA was detectable in the pL system. We modified both expression vectors to decrease the G + C content of the 5' end of the coding region without altering the predicted amino acid sequence. This resulted in expression of detectable G-CSF mRNA and protein in both systems. Expression reached 17% and 6.5% of the total soluble cellular protein in the pL and trpP expression systems, respectively. The N-terminal sequence of the recombinant G-CSF from the pL system was Met-Thr-Pro-Leu-Gly-Pro-. G-CSF isolated from several human cell lines (including the LD-1 cell line reported here), does not have an N-terminal methionyl residue. Deletion of the threonine codon at the beginning of the coding region for the mature G-CSF resulted in efficient removal of the N-terminal methionine residue during expression in E. coli. 相似文献
17.
Valentine AM LeTadic-Biadatti MH Toy PH Newcomb M Lippard SJ 《The Journal of biological chemistry》1999,274(16):10771-10776
Radical clock substrate probes were used to assess the viability of a discrete substrate radical species in the mechanism of hydrocarbon oxidation by the soluble methane monooxygenase (sMMO) from Methylococcus capsulatus (Bath). New substituted cyclopropane probes were used with very fast ring-opening rate constants and other desirable attributes, such as the ability to discriminate between radical and cationic intermediates. Oxidation of these substrates by a reconstituted sMMO system resulted in no rearranged products, allowing an upper limit of 150 fs to be placed on the lifetime of a putative radical species. This limit strongly suggests that there is no such substrate radical intermediate. The two enantiomers of trans-1-methyl-2-phenyl-cyclopropane were prepared, and the regioselectivity of their oxidation to the corresponding cyclopropylmethanol and cyclopropylphenol products was determined. The results are consistent with selective orientation of the two enantiomeric substrates in the hydrophobic cavity at the active site of sMMO, specific models for which were examined by molecular modeling. 相似文献
18.
Evaluation of procedures for amplification of small-size samples for hybridization on microarrays 总被引:3,自引:0,他引:3
Various approaches have been developed for the preparation of samples for gene expression monitoring. For Affymetrix chips, a standard protocol is widely used; however, this is inefficient for small samples such as laser capture microdissections. Several amplification procedures for such samples already exist, and our goal was to test two of them: the first is based on random PCR amplification, and the second, linear amplification, involves performing the standard protocol twice. We analyzed a dilution of a commercially available mouse brain total RNA preparation and microdissections from mouse hippocampus and striatum. We evaluated the quality of microarray data by analyzing several chip parameters and performing multiple comparisons. At the biological level, brain microdissections prepared with either method gave similar expression results. At the technical level, analysis of the commercial sample showed that random PCR amplification is more reproducible, requires smaller RNA input, and generates cRNA of higher quality than linear amplification. 相似文献
19.
This study was designed to measure the effect of iron supplementation on antioxidant status in iron-deficient anemia, including
the time for hemoglobin normalization and at the time of filling of iron body stores. The extent of plasma lipid peroxidation
was evaluated by measuring the levels of malondialdehyde and glutathione peroxidase (GSH-Px), and the activities of superoxide
dismutase (SOD) and catalase in 63 patients with iron-deficiency anemia before and after 6 wk of iron supplementation and
at the time when body iron stores are saturated. After 6 wk of iron supplementation, a significant decrease of oxidative stress
was observed in the treated subjects relative to controls (p<0.05). No significant differences existed between treated patients at 6 wk and at the end of the study. The erythrocyte levels
of catalase, SOD, and GSH-Px were significantly lower in treated patients relative to controls (p<0.05). These levels increased after 6 wk of supplementation (p<0.05) and showed no significant differences with those at the end of the study. 相似文献
20.
Omer Ateş Levent Dalyan Gulen Hatemi Vedat Hamuryudan Aysegul Topal-Sarıkaya 《Molecular biology reports》2010,37(7):3637-3641
We aim to ascertain the possible involvement of functional IL10 and TNF-α promoter polymorphisms on the susceptibility to Behçet’s syndrome (BS), to examine whether IL10 and TNF-α genotypes might work synergistically influencing susceptibility to BS. IL10 ?1082G/A, ?819C/T and ?592C/A and TNF ?308G/A polymorphisms were analyzed in 102 Turkish patients with BS and 102 healthy subjects by using amplification refractory mutation system-polymerase chain reaction (ARMS-PCR). We have found no significant associations between IL10 ?1082G/A, ?819C/T, ?592C/A, TNF-α ?308G/A polymorphisms and BS. Also, no significant correlation was found between IL10 GCC, ACC, ATA haplotypes, GCC+/GCC+, GCC+/GCC?, GCC?/GCC? genotypes. There was no significant association between combined TNF-α/IL10 genotypes and BS. Our study indicates that functional TNF-α, IL10 genotypes or combined TNF-α, IL10 genotypes do not play a role in BS susceptibility in Turkish BS patients. 相似文献