全文获取类型
收费全文 | 1506篇 |
免费 | 43篇 |
出版年
2023年 | 25篇 |
2022年 | 28篇 |
2021年 | 47篇 |
2020年 | 32篇 |
2019年 | 48篇 |
2018年 | 39篇 |
2017年 | 42篇 |
2016年 | 51篇 |
2015年 | 49篇 |
2014年 | 96篇 |
2013年 | 110篇 |
2012年 | 112篇 |
2011年 | 151篇 |
2010年 | 92篇 |
2009年 | 68篇 |
2008年 | 86篇 |
2007年 | 85篇 |
2006年 | 84篇 |
2005年 | 78篇 |
2004年 | 68篇 |
2003年 | 61篇 |
2002年 | 47篇 |
2001年 | 9篇 |
2000年 | 8篇 |
1999年 | 3篇 |
1998年 | 6篇 |
1997年 | 2篇 |
1996年 | 1篇 |
1995年 | 2篇 |
1994年 | 3篇 |
1993年 | 1篇 |
1991年 | 1篇 |
1990年 | 4篇 |
1989年 | 2篇 |
1987年 | 1篇 |
1986年 | 3篇 |
1982年 | 1篇 |
1979年 | 1篇 |
1964年 | 1篇 |
1963年 | 1篇 |
排序方式: 共有1549条查询结果,搜索用时 15 毫秒
81.
Stöcker M Tur MK Sasse S Krüssmann A Barth S Engert A 《Protein expression and purification》2003,28(2):211-219
Immunotoxins consist of a target-cell-specific binding moiety, chemically or recombinantly linked to a cytotoxic component. A number of different immunotoxins (IT) have increasingly been evaluated for immunotherapy. Since these foreign proteins are highly immunogenic in human, we have developed recombinant IT using the human ribonuclease angiogenin. Due to their potential toxic effects on eucaryotic cells, these IT are usually expressed in bacteria. Depending on the structure, size, and sequence of the desired IT, bacterial expression can be limited and the yield after purification be unsatisfactory. Therefore, the expression of IT in eucaryotic cells could provide a promising alternative. For this purpose we genetically fused the anti-CD30 single-chain variable fragment (scFv) Ki4 to the N- and C-termini of recombinant angiogenin. Both IT possess leader sequences, which mediate their secretion into the cell culture supernatant. Using a bicistronic mRNA the IT were simultaneously expressed together with enhanced green fluorescent protein (EGFP). This allows direct monitoring of transfected cells. An additional plasmid encoded Zeocin resistance enhances the cultivation of transfected cells under selection pressure. Three days after transfection of 293T-cells, unpurified IT were analyzed by flow cytometry and competitive cell proliferation assays. This is the first report on the use of eucaryotic cells for the secretion of functionally active IT with a human effector domain. 相似文献
82.
There are two highly homologous survival motor neuron (SMN) genes in humans but molecular defects in the SMN1 gene cause spinal muscular atrophy (SMA). More than 90% of SMA patients are shown to have a homozygous deletion of exon 7 in the SMN1 gene. Therefore, a simple test for exon 7 deletion would be very useful in the molecular diagnosis of SMA. However, limited methods are available, and most of these methods utilize expensive instruments and consumables. Here, we describe a simple allele-specific PCR test, which can be performed using standard equipment in DNA laboratories. The principle of the test is based on a single nucleotide difference (C versus T) between the exon 7 of SMN1 and SMN2 genes. Using allele-specific primers, two PCR amplifications are performed for each sample to amplify a 404-bp diagnostic fragment, and consequent electrophoresis of PCR products on agarose gel provides definitive information concerning the exon 7 deletion To rule out false negatives, a 500-bp fragment from the N-acetyltransferase gene was coamplified as an internal control in each test. We have, so far, analyzed 41 SMA samples with our method, and tested the validity of results using an independent restriction fragment length polymorphism (RFLP) method. Genotyping results obtained by both methods were in complete agreement for all of the samples analyzed. Our method can also be used to detect heterozygous deletion of exon 7 in SMN genes, if the relative intensities of the diagnostic and internal control bands are determined. 相似文献
83.
Ciftçi M Demir Y Ozmen I Atici O 《Journal of enzyme inhibition and medicinal chemistry》2003,18(1):71-76
The present study was undertaken to determine in vivo and in vitro effects of some plant growth regulators on rat erythrocyte carbonic anhydrase (CA) and glucose-6-phosphate dehydrogenase (G6PD) activities. Both in vivo and in vitro, spermidine and kinetin did not affect enzymatic activities of CA and G6PD, whereas putrescine decreased these activities, and abscisic acid increased them. Since plants use such growth regulators, their effects should be considered on mammals consuming them since they may possess important biological effects. 相似文献
84.
Whole body exposure of rats to microwaves emitted from a cell phone does not affect the testes 总被引:7,自引:0,他引:7
Dasdag S Zulkuf Akdag M Aksen F Yilmaz F Bashan M Mutlu Dasdag M Salih Celik M 《Bioelectromagnetics》2003,24(3):182-188
The objective of this study was to investigate the effects of radiofrequency radiation emitted from cellular phones on the lipid composition, malondialdehyde concentration, p53 immune reactivity, sperm count, morphology, histological structure of testes, and on rectal temperature of rats exposed to microwave radiation emitted from cellular phones. Sixteen Spraque-Dawley rats were separated into two groups of eight, sham exposed (control) and experimental. The rats were confined in plexiglas cages specially designed for this study, and cellular phones were placed 0.5 cm under the cages. For the experimental group, cellular phones were activated 20 min per day (7 days a week) for 1 month. For the control group, the cellular phones were placed beneath the cages for 20 min a day, but the phones were turned off. Rectal temperatures were measured weekly. For 250 mW radiated power, the whole body average SAR (rms) is 0.52 W/kg and 1 g averaged peak SAR (rms) is 3.13 W/kg. The Mann-Whitney U-test was used for statistical comparisons of groups. No statistically significant alteration in any of the endpoints was noted. This study found no evidence suggesting an adverse effect of cell phone exposure on measures of testicular function or structure. 相似文献
85.
Melatonin reduces glial reactivity in the hippocampus, cortex, and cerebellum of streptozotocin-induced diabetic rats 总被引:5,自引:0,他引:5
Baydas G Reiter RJ Yasar A Tuzcu M Akdemir I Nedzvetskii VS 《Free radical biology & medicine》2003,35(7):797-804
Hyperglycemia plays a critical role in the development and progression of diabetic neuropathy. One of the mechanisms by which hyperglycemia causes neural degeneration is via the increased oxidative stress that accompanies diabetes. Metabolic and oxidative insults often cause rapid changes in glial cells. Key indicators of this response are increased synthesis of glial fibrillary acidic protein (GFAP) and S100B, both astrocytic markers. In the present study, we examined glial reactivity in hippocampus, cortex, and cerebellum of streptozotocin (STZ)-induced diabetic rats by determining the expression of GFAP and S-100B and we evaluated the effect of melatonin on the glial response. Western blot measurement of contents in brain regions after 6 weeks of STZ-induced diabetes indicated significant increases in these constituents compared with those in nondiabetic controls. Administration of melatonin prevented the upregulation of GFAP in all brain regions of diabetic rats. Using GFAP immunohistochemistry, we observed an increase in GFAP immunostaining in the hippocampus of STZ-diabetic rats relative to levels in the control brains. Treatment with melatonin resulted in an obvious reduction of GFAP-immunoreactive astrocytes in hippocampus. Like GFAP, S100B levels also were increased in all three brain areas of diabetic rats, an effect also reduced by melatonin treatment. Finally, the levels of lipid peroxidation products were elevated as a consequence of diabetes, with this change also being prevented by melatonin. These results suggest that diabetes causes increased glial reactivity possibly due to elevated oxidative stress, and administration of melatonin represents an achievable adjunct therapy for preventing gliosis. 相似文献
86.
Serum cytokine levels in patients with acute brucellosis and their relation to the traditional inflammatory markers 总被引:2,自引:0,他引:2
Demirdag K Ozden M Kalkan A Godekmerdan A Sirri Kilic S 《FEMS immunology and medical microbiology》2003,39(2):149-153
The number of clinical studies on gamma interferon (IFN-gamma), tumor necrosis factor-alpha (TNF-alpha) and interleukin-4 (IL-4) in human brucellosis is limited. The present study was focused on IFN-gamma, TNF-alpha and IL-4 levels in acute brucellosis cases, in the acute phase and at the end of the treatment. The relation of these cytokines to traditional inflammation markers was also investigated. The study included 27 cases of acute brucellosis and 20 healthy volunteers who had no complaints. It was found that mean IFN-gamma and TNF-alpha levels, CRP (C-reactive protein) levels and ESR (erythrocyte sedimentation rate) values were significantly higher in acute brucellosis cases as compared to post-treatment values and values measured in the control group. In addition, IFN-gamma and TNF-alpha levels measured in the acute phase correlated with the increase in CRP levels and ESR values. Our results confirmed that IFN-gamma and TNF-alpha are involved in the pathophysiology of brucellosis and are closely related to the inflammatory activation of the disease. In view of the present findings, it is suggested that IFN-gamma and TNFalpha may be used for monitoring brucellosis. 相似文献
87.
Branched chain amino acids induce apoptosis in neural cells without mitochondrial membrane depolarization or cytochrome c release: implications for neurological impairment associated with maple syrup urine disease 总被引:6,自引:0,他引:6
下载免费PDF全文
![点击此处可从《Molecular biology of the cell》网站下载免费的PDF全文](/ch/ext_images/free.gif)
Jouvet P Rustin P Taylor DL Pocock JM Felderhoff-Mueser U Mazarakis ND Sarraf C Joashi U Kozma M Greenwood K Edwards AD Mehmet H 《Molecular biology of the cell》2000,11(5):1919-1932
Maple syrup urine disease (MSUD) is an inborn error of metabolism caused by a deficiency in branched chain alpha-keto acid dehydrogenase that can result in neurodegenerative sequelae in human infants. In the present study, increased concentrations of MSUD metabolites, in particular alpha-keto isocaproic acid, specifically induced apoptosis in glial and neuronal cells in culture. Apoptosis was associated with a reduction in cell respiration but without impairment of respiratory chain function, without early changes in mitochondrial membrane potential and without cytochrome c release into the cytosol. Significantly, alpha-keto isocaproic acid also triggered neuronal apoptosis in vivo after intracerebral injection into the developing rat brain. These findings suggest that MSUD neurodegeneration may result, at least in part, from an accumulation of branched chain amino acids and their alpha-keto acid derivatives that trigger apoptosis through a cytochrome c-independent pathway. 相似文献
88.
89.
90.
Dilara Kaman Necip İlhan Kerem Metin Mehmet Akbulut Bilal Üstündağ 《Cell biochemistry and function》2009,27(2):88-92
Paraoxonase 1 (PON 1) is a high‐density lipoprotein (HDL)‐associated enzyme with antioxidant function protecting low‐density lipoprotein (LDL) from oxidation. PON 1 has two amino acid polymorphisms in coding region; L/M 55 and Q/R 192. These polymorphisms modulate paraoxonase activity of the enzyme. PON 1 activity decreases in coronary artery disease (CAD). In the present study, distribution of PON 1 L/M 55 and Q/R 192 polymorphisms and the effect of these polymorphisms on the activities of PON 1, and on the severity of CAD in 277 CAD (+) patient and 92 CAD (?) subjects were examined. PON 1 L/M 55 and Q/R 192 genotypes were determined by PCR, RFLP and agarose gel electrophoresis techniques. Genotype distributions and allele frequencies for PON 1 Q/R 192 polymorphism were not significantly different between controls and CAD (+) patient group (p > 0.05), but in genotype and allele distribution of PON 1 L/M55 polymorphism, there was significantly difference among groups (p < 0.05). Genotype distributions for both polymorphisms were not significantly different between subgroups of single‐vessel disease (SVD), double‐vessel disease (DVD) and triple‐vessel disease (TVD). Serum PON 1 activity was lower in CAD (+) group than in controls and this was also statistically significant (p < 0.001). In both groups, the highest PON activities were detected in LL and RR genotypes. In summary, our results suggest that there is an association between the PON 1 L/M 55 polymorphism of paraoxonase and CAD in Turkish patients but not with PON 1 Q/R 192 polymorphism. However, it is hard to correlate these polymorphisms and severity of CAD. Copyright © 2009 John Wiley & Sons, Ltd. 相似文献