全文获取类型
收费全文 | 894篇 |
免费 | 57篇 |
国内免费 | 2篇 |
出版年
2024年 | 1篇 |
2023年 | 2篇 |
2022年 | 6篇 |
2021年 | 12篇 |
2020年 | 8篇 |
2019年 | 19篇 |
2018年 | 11篇 |
2017年 | 21篇 |
2016年 | 28篇 |
2015年 | 28篇 |
2014年 | 22篇 |
2013年 | 47篇 |
2012年 | 75篇 |
2011年 | 74篇 |
2010年 | 50篇 |
2009年 | 40篇 |
2008年 | 65篇 |
2007年 | 52篇 |
2006年 | 41篇 |
2005年 | 43篇 |
2004年 | 41篇 |
2003年 | 41篇 |
2002年 | 43篇 |
2001年 | 17篇 |
2000年 | 10篇 |
1999年 | 23篇 |
1998年 | 12篇 |
1997年 | 8篇 |
1996年 | 7篇 |
1995年 | 8篇 |
1994年 | 6篇 |
1993年 | 7篇 |
1992年 | 7篇 |
1991年 | 16篇 |
1990年 | 9篇 |
1989年 | 7篇 |
1988年 | 3篇 |
1987年 | 6篇 |
1986年 | 5篇 |
1985年 | 5篇 |
1984年 | 3篇 |
1983年 | 3篇 |
1982年 | 6篇 |
1981年 | 4篇 |
1980年 | 3篇 |
1979年 | 4篇 |
1977年 | 1篇 |
1976年 | 1篇 |
1973年 | 1篇 |
1958年 | 1篇 |
排序方式: 共有953条查询结果,搜索用时 15 毫秒
81.
Shuhei Yanase Ryosuke Yamada Shohei Kaneko Hideo Noda Tomohisa Hasunuma Tsutomu Tanaka Chiaki Ogino Hideki Fukuda Akihiko Kondo Professor 《Biotechnology journal》2010,5(5):449-455
We demonstrate direct ethanol fermentation from amorphous cellulose using cellulase-co-expressing yeast. Endoglucanases (EG) and cellobiohydrolases (CBH) from Trichoderma reesei, and β-glucosidases (BGL) from Aspergillus aculeatus were integrated into genomes of the yeast strain Saccharomyces cerevisiae MT8-1. BGL was displayed on the yeast cell surface and both EG and CBH were secreted or displayed on the cell surface. All enzymes were successfully expressed on the cell surface or in culture supernatants in their active forms, and cellulose degradation was increased 3- to 5-fold by co-expressing EG and CBH. Direct ethanol fermentation from 10 g/L phosphoric acid swollen cellulose (PASC) was also carried out using EG-, CBH-, and BGL-co-expressing yeast. The ethanol yield was 2.1 g/L for EG-, CBH-, and BGL-displaying yeast, which was higher than that of EG- and CBH-secreting yeast (1.6 g/L ethanol). Our results show that cell surface display is more suitable for direct ethanol fermentation from cellulose. 相似文献
82.
83.
Koichi Murata Hiroyuki Yoshitomi Shimei Tanida Masahiro Ishikawa Kohei Nishitani Hiromu Ito Takashi Nakamura 《Arthritis research & therapy》2010,12(3):R86
Introduction
MicroRNAs (miRNAs), endogenous small noncoding RNAs regulating the activities of target mRNAs and cellular processes, are present in human plasma in a stable form. In this study, we investigated whether miRNAs are also stably present in synovial fluids and whether plasma and synovial fluid miRNAs could be biomarkers of rheumatoid arthritis (RA) and osteoarthritis (OA). 相似文献84.
Koseki M Hirano K Masuda D Ikegami C Tanaka M Ota A Sandoval JC Nakagawa-Toyama Y Sato SB Kobayashi T Shimada Y Ohno-Iwashita Y Matsuura F Shimomura I Yamashita S 《Journal of lipid research》2007,48(2):299-306
Lipid rafts on the cell surface are believed to be very important as platforms for various cellular functions. The aim of this study was to know whether defective lipid efflux may influence lipid rafts on the cell surface and their related cellular functions. We investigated macrophages with defective lipid efflux from ATP binding cassette transporter A1-deficient (Abca1-KO) mice. Lipid rafts were evaluated by the following two novel probes: a biotinylated and protease (subtilisin Carlsberg)-nicked derivative of theta-toxin and a fluorescein ester of polyethylene glycol-derived cholesterol. Lipid rafts in Abca1-KO macrophages were increased, as demonstrated by both probes. Moreover, activities of nuclear factor kappaB, mRNA and intracellular distribution, and secretion of tumor necrosis factor-alpha (TNF-alpha) were examined after stimulation by lipopolysaccharides (LPSs). LPS-induced responses of the activation of nuclear factor kappaB and TNF-alpha were more prompt and accelerated in the Abca1-KO macrophages compared with wild-type macrophages. Modification of lipid rafts by cyclodextrin and nystatin corrected the abnormal response, suggesting an association between the increased lipid rafts and abnormal TNF-alpha secretion. We report here that Abca1-KO macrophages with defective lipid efflux exhibited increased lipid rafts on the cell surface and accelerated TNF-alpha secretion. 相似文献
85.
Kimura MY Iwamura C Suzuki A Miki T Hasegawa A Sugaya K Yamashita M Ishii S Nakayama T 《Journal of immunology (Baltimore, Md. : 1950)》2007,178(8):4926-4936
Schnurri-2 (Shn-2) is a large zinc-finger containing protein, and it plays a critical role in cell growth, signal transduction and lymphocyte development. In Shn-2-deficient CD4 T cells, the activation of NF-kappaB was up-regulated and their ability to differentiate into Th2 cells was enhanced. We herein demonstrate that Th1 and Th2 memory cells are not properly generated from Shn-2-deficient effector Th1/Th2 cells. Even a week after the transfer of effector Th1/Th2 cells into syngeneic mice, a dramatic decrease in the number of Shn-2-deficient donor T cells was detected particularly in the lymphoid organs. The transferred Shn-2-deficient Th1/Th2 cells express higher levels of the activation marker CD69. No significant defect in the BrdU incorporation in the Shn-2-deficient transferred CD4 T cells was observed. The numbers of apoptotic cells were selectively higher in Shn-2-deficient donor Th1/Th2 cell population. Moreover, Shn-2-deficient effector Th1 and Th2 cells showed an increased susceptibility to cell death in in vitro cultures with increased expression of FasL. Transfer of Th2 effector cells over-expressing the p65 subunit of NF-kappaB resulted in a decreased number of p65-expressing cells in the lymphoid organs. As expected, T cell-dependent Ab responses after in vivo immunization of Shn-2-deficient mice were significantly reduced. Thus, Shn-2 appears to control the generation of memory Th1/Th2 cells through a change in their susceptibility to cell death. 相似文献
86.
The ras proto-oncogenes, of which there are four isoforms, are molecular switches that function in signal transduction pathways to control cell differentiation, proliferation, and survival. How the Ras isoforms orchestrate cellular processes that affect behavior is poorly understood. Further, why cells express two or more Ras isoforms is unknown. Here, using a genetically defined system, we show that the presence of both wild-type KRas and NRas isoforms is required for transformation because they perform distinct nonoverlapping functions: wild-type NRas regulates adhesion, and KRas coordinates motility. Remarkably, we find that Ras isoforms achieve functional specificity by engaging different signaling pathways to affect the same cellular processes, thereby coordinating cellular outcome. Although we find that signaling from both isoforms intersects in actin and microtubule cytoskeletons, our results suggest that KRas signals through Akt and Cdc42 while NRas signals through Raf and RhoA. Our analyses suggest a previously unappreciated convergence of different Ras isoforms on the dynamics of the processes involved in transformation. 相似文献
87.
The extracellular matrix (ECM) acts as a critical factor during morphogenesis. Because the organization of the ECM directly
influences the structure of tissues and organs, a determination of the way that ECM organization is regulated should help
to clarify morphogenesis. We have analyzed the assembly of Del1, an ECM protein produced by endothelial cells in embryos,
in the ECM. Del1 consists of three epidermal growth factor repeats (E1–E3) at its N-terminus and two discoidin domains (C1,
C2) at its C-terminus. Experiments with various deletion mutants of Del1 have revealed that fragments containing the C-terminus
of C1, which has a lectin-like structure, direct deposition in the ECM. The efficiency of deposition varies according to the
presence of other domains in Del1. A fragment containing E3 and C1 has the strongest deposition activity, whereas fragments
containing C2, which is highly homologous to C1, have low deposition activity. Digestion of ECM with hyaluronidase from bovine
testis releases Del1 from the ECM, suggesting that glycosaminoglycans are involved in the deposition of Del1. In vivo gene
transfer experiments have shown that fusion with the deposition domain of Del1 dramatically alters the distribution of exogenous
proteins in mice. Thus, the extent of Del1 deposition may modify the organization of the ECM. 相似文献
88.
Fujiki T Nanatani K Nishitani K Yagi K Ohnishi F Yoneyama H Uchida T Nakajima T Abea K 《Journal of biochemistry》2007,141(1):85-91
We cloned the aspT gene encoding the L-aspartate:L-alanine antiporter AspTCt in Comamonas testosteroni genomic DNA. Analysis of the nucleotide sequence revealed that C. testosteroni has an asp operon containing aspT upstream of the l-aspartate 4-decarboxylase gene, and that the gene order of the asp operon of C. testosteroni is the inverse of that of Tetragenococcus halophilus. We used proteoliposomes to confirm the transport processes of AspTCt. To elucidate the two-dimensional structure of AspTCt, we analysed its membrane topology by means of alkaline phosphatase (PhoA) and beta-lactamase (BlaM) fusion methods. The fusion analyses revealed that AspTCt has seven transmembrane segments (TMs), a large cytoplasmic loop containing approximately 200 amino acid residues between TM4 and TM5, a cytoplasmic N-terminus, and a periplasmic C-terminus. These results suggest that the orientation of the N-terminus of AspTCt differs from that of tetragenococcal AspT, even though these two AspT orthologues catalyse the same transport reactions. 相似文献
89.
Nishizawa R Nishiyama T Hisaichi K Matsunaga N Minamoto C Habashita H Takaoka Y Toda M Shibayama S Tada H Sagawa K Fukushima D Maeda K Mitsuya H 《Bioorganic & medicinal chemistry letters》2007,17(3):727-731
Hydroxylated derivatives were designed and synthesized based on the information of oxidative metabolites. Compounds derived from beta-substituted (2R,3R)-2-amino-3-hydroxypropionic acid showed improved inhibitory activities against the binding of MIP-1alpha to human CCR5, compared with the non-hydroxylated derivatives and the other isomers. 相似文献
90.
Molecular survey of Babesia microti, Ehrlichia species and Candidatus neoehrlichia mikurensis in wild rodents from Shimane Prefecture, Japan 总被引:1,自引:0,他引:1
Tabara K Arai S Kawabuchi T Itagaki A Ishihara C Satoh H Okabe N Tsuji M 《Microbiology and immunology》2007,51(4):359-367
A significant number of patients are diagnosed with "fevers of unknown origin" (FUO) in Shimane Prefecture in Japan where tick-borne diseases are endemic. We conducted molecular surveys for Babesia microti, Ehrlichia species, and Candidatus Neoehrlichia mikurensis in 62 FUO cases and 62 wild rodents from Shimane Prefecture, Japan. PCR using primers specific for the Babesia 18S small-subunit rRNA (rDNA) gene and Anaplasmataceae groESL amplified products from 45% (28/62) and 25.8% (16/62) of captured mice, respectively. Of the 28 18S rDNA PCR positives, 23 and five samples were positive for Hobetsu- and Kobe-type B. microti, respectively. In contrast, of the 16 groESL PCR positives, eight, one and seven samples were positive for Ehrlichia muris, Ehrlichia sp. HF565 and Candidatus N. mikurensis, respectively. Inoculation of selected blood samples into Golden Syrian hamsters indicated the presence of Hobetsu- and Kobe-type B. microti in four and one sample, respectively. Isolation of the latter strain was considered important as previous studies suggested that the distribution of this type was so far confined to Awaji Island in Hyogo Prefecture, where the first case of transfusion-associated human babesiosis originated. DNA samples from 62 FUO human cases tested negative for B. microti 18S rDNA gene, Anaplasmataceae groESL gene, Rickettsia japonica 17K genus-common antigen gene and Orientia tsutsugamushi 56K antigen gene by PCRs. We also conducted seroepidemiological surveys on 62 human sera collected in Shimane Prefecture from the FUO patients who were suspected of carrying tick-borne diseases. However, indirect immunofluorescent antibody tests using B. microti- and E. muris-infected cells detected IgG against E. muris in only a single positive sample. This study demonstrates the presence of several potentially important tick-borne pathogens in Shimane Prefecture and suggests the need for further study on the causative agents of FUOs. 相似文献