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101.
Foxp3 inhibits RORgammat-mediated IL-17A mRNA transcription through direct interaction with RORgammat 总被引:3,自引:0,他引:3
Ichiyama K Yoshida H Wakabayashi Y Chinen T Saeki K Nakaya M Takaesu G Hori S Yoshimura A Kobayashi T 《The Journal of biological chemistry》2008,283(25):17003-17008
The cytokine, transforming growth factor-beta1 (TGF-beta1), converts naive T cells into regulatory T cells that prevent autoimmunity. However, in the presence of interleukin (IL)-6, TGF-beta1 has also been found to promote differentiation into IL-17-producing helper T (Th17) cells that are deeply involved in autoimmunity and inflammation. However, it has not been clarified how TGF-beta1 and IL-6 determine such a distinct fate. Here we found that a master regulator for Th17, retinoic acid-related orphan receptor gammat (RORgammat), was rapidly induced by TGF-beta1 regardless of the presence of IL-6. IL-6 reduced Foxp3 expression, and overexpression of Foxp3 in a T cell line resulted in a strong reduction of IL-17A expression. We have characterized the IL-17A promoter and found that RORgammat binding is sufficient for activation of the minimum promoter in the HEK 293T cells. RORgammat-mediated IL-17A promoter activation was suppressed by forced expression of Foxp3. Foxp3 directly interacted with RORgammat through exon 2 region of Foxp3. The exon 2 region and forkhead (FKH) domain of Foxp3 were necessary for the suppression of RORgammat-mediated IL-17A promoter activation. We propose that induction of Foxp3 is the mechanism for the suppression of Th17 and polarization into inducible Treg. 相似文献
102.
Takeshi Fukuda Seizaburo Shiraga Michiko Kato Shohei Yamamura Yasutaka Morita Eiichi Tamiya Teruo Hori Shin-ichiro Suye Prof./Dr. Mitsuyoshi Ueda 《NanoBioTechnology》2005,1(1):105-111
A novel screening system using a microchamber array chip was developed for construction of combinatorial nano-sized protein
libraries in combination with yeast cell surface engineering. It is possible to place a single yeast cell into each microchamber,
to observe its behavior, and to pick up the target cell. The microchamber array chip is referred to as a “yeast cell chip.”
A single EGFP-displaying yeast cell could be detected, picked up by a micro-manipulator, and cultivated on agar medium. Furthermore,
a catalytic reaction, the hydrolysis of fluorescein dioctanate, by a single yeast cell displaying Rhizopus oryzae lipase (ROL) was carried out in one microchamber. The ROL-encoding gene in a single ROL-displaying cell was amplified by
PCR. These results demonstrate that this yeast cell chip in combination with cell surface engineering could be used as a tool
in a high-throughput screening system not only for a single living cell and a whole-cell catalyst with a nano-sized protein
cluster but also for modified nano-sized and functional protein molecules from protein libraries on the cell surface. 相似文献
103.
Coaggregation among Nonflocculating Bacteria Isolated from Activated Sludge 总被引:7,自引:3,他引:4 下载免费PDF全文
Anushree Malik Masashi Sakamoto Shohei Hanazaki Masamitsu Osawa Takanori Suzuki Masaki Tochigi Kazuo Kakii 《Applied microbiology》2003,69(10):6056-6063
Thirty-two strains of nonflocculating bacteria isolated from sewage-activated sludge were tested by a spectrophotometric assay for their ability to coaggregate with one other in two-membered systems. Among these strains, eight showed significant (74 to 99%) coaggregation with Acinetobacter johnsonii S35 while only four strains coaggregated, to a lesser extent (43 to 65%), with Acinetobacter junii S33. The extent and pattern of coaggregation as well as the aggregate size showed good correlation with cellular characteristics of the coaggregating partners. These strains were identified by sequencing of full-length 16S rRNA genes. A. johnsonii S35 could coaggregate with strains of several genera, such as Oligotropha carboxidovorans, Microbacterium esteraromaticum, and Xanthomonas spp. The role of Acinetobacter isolates as bridging organisms in multigeneric coaggregates is indicated. This investigation revealed the role of much-neglected nonflocculating bacteria in floc formation in activated sludge. 相似文献
104.
Natsumi Takei Takuma Nakamura Shohei Kawamura Yuki Takada Yui Satoh Atsushi P. Kimura Tomoya Kotani 《Biological procedures online》2018,20(1):6
Background
Subcellular localization of coding and non-coding RNAs has emerged as major regulatory mechanisms of gene expression in various cell types and many organisms. However, techniques that enable detection of the subcellular distribution of these RNAs with high sensitivity and high resolution remain limited, particularly in vertebrate adult tissues and organs. In this study, we examined the expression and localization of mRNAs encoding Pou5f1/Oct4, Mos, Cyclin B1 and Deleted in Azoospermia-like (Dazl) in zebrafish and mouse ovaries by combining tyramide signal amplification (TSA)-based in situ hybridization with paraffin sections which can preserve cell morphology of tissues and organs at subcellular levels. In addition, the distribution of a long non-coding RNA (lncRNA), lncRNA-HSVIII, in mouse testes was examined by the same method.Results
The mRNAs encoding Mos, Cyclin B1 and Dazl were found to assemble into distinct granules that were distributed in different subcellular regions of zebrafish and mouse oocytes, suggesting conserved and specific regulations of these mRNAs. The lncRNA-HSVIII was first detected in the nucleus of spermatocytes at prophase I of the meiotic cell cycle and was then found in the cytoplasm of round spermatids, revealing expression patterns of lncRNA during germ cell development. Collectively, the in situ hybridization method demonstrated in this study achieved the detection and comparison of precise distribution patterns of coding and non-coding RNAs at subcellular levels in single cells of adult tissues and organs.Conclusions
This high-sensitivity and high-resolution in situ hybridization is applicable to many vertebrate species and to various tissues and organs and will be useful for studies on the subcellular regulation of gene expression at the level of RNA localization.105.
106.
"Flipper rubbing" behavior was quantitatively analyzed in wild Indo-Pacific bottlenose dolphins ( Tursiops aduncus ) around Mikura Island, Tokyo, Japan. We observed two types of flipper rubbing: (1) F-B rubbing; one dolphin (Rubber) rubbed its flipper over various parts of a partner's (Rubbee) body, and (2) F-F rubbing; both dolphins rubbed each other's anterior flipper edge in alternating shifts. F-B rubbings tended to be initiated by the Rubbee and were terminated by the Rubber. The Rubbee often moved actively its body part that was in contact with the Rubber's flipper, and assumed side-up, upside-down, or other postures while the Rubber remained horizontal in most cases. These facts suggest that the Rubbee engaged in F-B rubbing more actively than the Rubber, and might receive some benefit from the frictional contact during F-B rubbing. Dolphins often switched their roles as Rubber and Rubbee between episodes of flipper rubbing bout. Adults and sub-adults exchanged F-B rubbing and F-F rubbing most often with individuals of the same sex in the same age class. F-B rubbing was frequent in mother-and-calf dyads. Our results suggest that flipper rubbing is an affiliative behavior which could be a quantitative measure of social relationships among individuals of this species in future studies. 相似文献
107.
Katsuhiro Shiratake Yoshinori Kanayama Masayoshi Maeshima Shohei Yamaki 《Physiologia plantarum》1998,103(3):312-319
Protoplasts isolated from pear fruit at the end of the cell‐division stage, 30 days after flowering (DAF), had already formed a large central vacuole and the vacuole occupied most of the protoplast. The changes in protein composition and density of the tonoplast (vacuolar membrane) were investigated during fruit development. After a linear sucrose density gradient centrifugation, the distribution of tonoplasts at 30 and 48 DAF was broad and began to narrow with further fruit development. This suggests that the tonoplast of young fruit is heterogeneous and becomes homogeneous with fruit development. The apparent density of the tonoplast at 30 DAF was approximately 1.12 g ml−1 ; it decreased with fruit development and was finally 1.09 g ml−1 in mature fruit. The phospholipid amount on the basis of tonoplast protein was 0.80 mg mg−1 at 30 DAF. It increased with fruit development, and finally reached 7.49 mg mg−1 . This result indicates that the decrease in the density of the tonoplast was caused by the increase in the ratio of phospholipid to membrane protein. The protein composition of the tonoplast at each stage was quite different. The level of polypeptides of 94, 70, 61, 52, 48 and 41 kDa was low in young fruit and high in the middle or later stages of fruit development. In contrast, the level of a 76‐kDa polypeptide was high in young fruit and decreased with fruit development. Although their functions are still unclear, these tonoplast proteins may play important roles in fruit development. 相似文献
108.
Fusae Kanemitsu Shohei Kira 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》1999,721(2):95
High enzyme activity of mitochondrial creatine kinase (creatine-N-phosphotransferase, mCK, EC 2.7.3.2) was detected in serum from a patient with advanced carcinoma of the rectum and its isoforms were characterized by means of isoelectric focusing (IEF). Three forms of mCK, membrane-bound (pI 6.9–7.0), octameric (pI 7.0–7.9) and dimeric (pI 6.7, 6.8, 6.9 and 7.0), were detected in the fresh serum. These three forms of mCK were converted to five dimeric isoforms, and these were characterized as one reduced form (pI 7.0) and four oxidized (pI 6.6, 6.7, 6.8 and 6.9) forms upon treatment with urea, hydrogen peroxide or 2-mercaptoethanol (2-ME). The C-terminal of the mCKs was concluded to be a lysine residue because the mCKs treated with carboxypeptidase B migrated to positions closer to the anode than did those not treated with carboxypeptidase B. Therefore, four bands were concluded to represent one reduced-delysined isoform (pI 6.4) and three oxidized-delysined isoforms (pI 6.1, 6.2 and 6.3). The broad octameric mCK band disappeared and a narrow band focused at pI 6.8–6.9 appeared upon probable delysination of the mCKs. Thus, the number of lysine residues at the C-terminal of the octamer was concluded to be variable due to variable catalysis by carboxypeptidase N in the plasma. mCKs seemed to be inactivated during conversion from a membrane-bound form to dimeric oxidized-delysined forms via the octameric, dimeric reduced and oxidized forms. 相似文献
109.
Isolation of Vacuoles from Immature Apple Fruit Flesh and Compartmentation of Sugars, Organic Acids, Phenolic Compounds and Amino Acids 总被引:4,自引:0,他引:4
Vacuoles of immature apple fruit (Malus pumila Mill. var. domesticaSchneid.) were obtained by purification using Ficoll densitygradient centrifugation after lysis of the protoplasts by bothmild osmotic shock and the addition of EDTA and BSA. The recoverywas about 35% of the protoplasts. The isolated vacuoles hada mean diameter of about 100 µm. The distribution of sugars, organic acids, phenolic compoundsand amino acids in the vacuole, the cytoplasm and the free spacewas determined. Almost all of the fructose and glucose, themajor sugars of the tissue, were found in the vacuole. Sorbitolwas mainly located in the free space and the vacuole, and sucrosein the free space and the cytoplasm. More than 90% of the malicacid, the main organic acid, was located in the vacuole. Almostall of the phenolic compounds were also deposited in the vacuole. The volumes of the vacuole, the cytoplasm and the free spacein the whole tissue were calculated from the cell numbers ofthe whole tissue, the volume of the isolated protoplasts, andthe volume of the vacuoles present in the protoplast. The soluteconcentration in each compartment was estimated: vacuoles, 888mM; cytoplasm, 37 mM; free space, 57 mM. How these compartmentationsof solutes affected the translocation of sugars into the fruitand the cell expansion is discussed.
1This paper is contribution A-159 of the Fruit Tree ResearchStation. (Received July 7, 1983; Accepted November 14, 1983) 相似文献
110.
Kentaro Shirotsuki Yuji Nonaka Jiro Takano Keiichi Abe So-ichiro Adachi Shohei Adachi Mutsuhiro Nakao 《BioPsychoSocial medicine》2017,11(1):25