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81.
Yuki Kawai Takeshi Kikuchi Yasumasa Mitani Yasushi Kogo Masayoshi Itoh Kengo Usui Hajime Kanamori Ai Kaiho Hideki Takakura Kanako Hoshi Paul E Cizdziel Yoshihide Hayashizaki 《Biologicals》2008,36(4):234-238
In a previous study, a single nucleotide polymorphism (SNP) diagnostic system named the SMart Amplification Process version 2 (SMAP 2) was reported, which enabled rapid gene diagnostics from crude samples such as whole blood. The asymmetric primer design and use of Taq MutS were reported as innovative background suppression technologies employed by SMAP 2, but Taq MutS is known to display differential affinities for various mismatch combinations, and hence may not be entirely effective for all possible applications. To address this issue we developed another approach using a competitive probe (CP) to enhance background suppression technology instead of Taq MutS. CP is a 3'-end aminated oligonucleotide that competes with 3'-end of a discrimination primer or the self-priming elongation site on intermediate product 2 (IM2) for non-target sequences, such as the alternative allele. The preferred hybridization kinetics for the full-match CP on the non-target sequence results in effective background suppression in SMAP 2 assays. By using a CP, we demonstrated the sensitive detection of EGFR gene mutations in purified genomic DNA from mixed cell populations. The CP approach is another tool enhancing the effectiveness and versatility of SMAP 2 assays, expanding its potential applications, and reinforcing its position as a highly effective technology for molecular diagnostics. 相似文献
82.
83.
Hayashizaki Y 《Comptes rendus biologies》2003,326(10-11):923-929
The Riken mouse genome encyclopedia a comprehensive full-length cDNA collection and sequence database. High-level functional annotation is based on sequence homology search, expression profiling, mapping and protein-protein interactions. More than 1000000 clones prepared from 163 tissues were end-sequenced and classified into 128000 clusters, and 60000 representative clones were fully sequenced representing 24000 clear protein-encoding genes. The application of the mouse genome database for positional cloning and gene network regulation analysis is reported. 相似文献
84.
Yamashita T Deguchi K Sehara Y Lukic-Panin V Zhang H Kamiya T Abe K 《Neurochemical research》2009,34(4):707-710
Possible strategies for treating ischemic stroke include: (1) Neuroprotection: preventing damaged neurons from undergoing
apoptosis in the acute phase of cerebral ischemia; (2) Stem cell therapy: the repair of broken neuronal networks with newly
born neurons in the chronic phase of cerebral ischemia. Firstly, we studied the neuroprotective effect of a calcium channel
blocker, azelnidipine, or a by-product of heme degradation, biliverdin, in the ischemic brain. These results revealed both
azelnidipine and biliverdin had a neuroprotective effect in the ischemic brain through their anti-oxidative property. Secondly,
we investigated the role of granulocyte colony-stimulating factor (G-CSF) by administering G-CSF to rats after cerebral ischemia
and found G-CSF plays a critical role in neuroprotection. Lastly, we developed a restorative stroke therapy with a bio-affinitive
scaffold, which is able to provide an appropriate environment for newly born neurons. In the future, we will combine these
strategies to develop more effective therapies for treatment of strokes.
Special issue article in honor of Dr. Akitane Mori. 相似文献
85.
86.
Atsutaka Kubosaki Yasuhiro Tomaru Michihira Tagami Erik Arner Hisashi Miura Takahiro Suzuki Masanori Suzuki Harukazu Suzuki Yoshihide Hayashizaki 《Genome biology》2009,10(4):R41-14
Background
Immediate early genes are considered to play important roles in dynamic gene regulatory networks following exposure to appropriate stimuli. One of the immediate early genes, early growth response gene 1 (EGR-1), has been implicated in differentiation of human monoblastoma cells along the monocytic commitment following treatment with phorbol ester. EGR-1 has been thought to work as a modifier of monopoiesis, but the precise function of EGR-1 in monocytic differentiation has not been fully elucidated. 相似文献87.
88.
Hattori Y Asano T Niki Y Kondoh H Kirihata M Yamaguchi Y Wakamiya T 《Bioorganic & medicinal chemistry》2006,14(10):3258-3262
Magnetic resonance imaging (MRI) and boron-neutron capture therapy (BNCT) are quite attractive techniques for diagnosis and treatment of cancer, respectively. In order to progress the study on both MRI and BNCT, the novel compounds containing 19F and 10B atoms in a single molecule were designed and synthesized. In the present paper, the syntheses and the internalization rates into tumor cells of these compounds are elucidated. 相似文献
89.
Yoshikawa F Banno Y Otani Y Yamaguchi Y Nagakura-Takagi Y Morita N Sato Y Saruta C Nishibe H Sadakata T Shinoda Y Hayashi K Mishima Y Baba H Furuichi T 《PloS one》2010,5(11):e13932
Background
Phospholipase D (PLD) catalyzes conversion of phosphatidylcholine into choline and phosphatidic acid, leading to a variety of intracellular signal transduction events. Two classical PLDs, PLD1 and PLD2, contain phosphatidylinositide-binding PX and PH domains and two conserved His-x-Lys-(x)4-Asp (HKD) motifs, which are critical for PLD activity. PLD4 officially belongs to the PLD family, because it possesses two HKD motifs. However, it lacks PX and PH domains and has a putative transmembrane domain instead. Nevertheless, little is known regarding expression, structure, and function of PLD4.Methodology/Principal Findings
PLD4 was analyzed in terms of expression, structure, and function. Expression was analyzed in developing mouse brains and non-neuronal tissues using microarray, in situ hybridization, immunohistochemistry, and immunocytochemistry. Structure was evaluated using bioinformatics analysis of protein domains, biochemical analyses of transmembrane property, and enzymatic deglycosylation. PLD activity was examined by choline release and transphosphatidylation assays. Results demonstrated low to modest, but characteristic, PLD4 mRNA expression in a subset of cells preferentially localized around white matter regions, including the corpus callosum and cerebellar white matter, during the first postnatal week. These PLD4 mRNA-expressing cells were identified as Iba1-positive microglia. In non-neuronal tissues, PLD4 mRNA expression was widespread, but predominantly distributed in the spleen. Intense PLD4 expression was detected around the marginal zone of the splenic red pulp, and splenic PLD4 protein recovered from subcellular membrane fractions was highly N-glycosylated. PLD4 was heterologously expressed in cell lines and localized in the endoplasmic reticulum and Golgi apparatus. Moreover, heterologously expressed PLD4 proteins did not exhibit PLD enzymatic activity.Conclusions/Significance
Results showed that PLD4 is a non-PLD, HKD motif-carrying, transmembrane glycoprotein localized in the endoplasmic reticulum and Golgi apparatus. The spatiotemporally restricted expression patterns suggested that PLD4 might play a role in common function(s) among microglia during early postnatal brain development and splenic marginal zone cells. 相似文献90.
Junko Tokuno Tsuyoshi Shoji Ryota Sumitomo Yuichiro Ueda Keiji Yamanashi Cheng-long Huang 《World journal of surgical oncology》2017,15(1):212