全文获取类型
收费全文 | 816篇 |
免费 | 69篇 |
专业分类
885篇 |
出版年
2023年 | 4篇 |
2022年 | 11篇 |
2021年 | 14篇 |
2020年 | 8篇 |
2019年 | 5篇 |
2018年 | 21篇 |
2017年 | 6篇 |
2016年 | 15篇 |
2015年 | 28篇 |
2014年 | 25篇 |
2013年 | 57篇 |
2012年 | 57篇 |
2011年 | 56篇 |
2010年 | 28篇 |
2009年 | 28篇 |
2008年 | 41篇 |
2007年 | 36篇 |
2006年 | 51篇 |
2005年 | 54篇 |
2004年 | 40篇 |
2003年 | 54篇 |
2002年 | 46篇 |
2001年 | 21篇 |
2000年 | 16篇 |
1999年 | 18篇 |
1998年 | 9篇 |
1997年 | 7篇 |
1996年 | 12篇 |
1995年 | 8篇 |
1994年 | 14篇 |
1993年 | 6篇 |
1992年 | 7篇 |
1991年 | 6篇 |
1990年 | 4篇 |
1989年 | 10篇 |
1988年 | 9篇 |
1987年 | 5篇 |
1986年 | 7篇 |
1985年 | 8篇 |
1984年 | 4篇 |
1982年 | 5篇 |
1981年 | 3篇 |
1980年 | 4篇 |
1978年 | 2篇 |
1977年 | 2篇 |
1976年 | 2篇 |
1974年 | 4篇 |
1973年 | 2篇 |
1972年 | 2篇 |
1971年 | 1篇 |
排序方式: 共有885条查询结果,搜索用时 0 毫秒
21.
In vitro phosphorylation of the tumor suppressor gene RB protein by mitosis-specific histone H1 kinase 总被引:19,自引:0,他引:19
Y Taya H Yasuda M Kamijo K Nakaya Y Nakamura Y Ohba S Nishimura 《Biochemical and biophysical research communications》1989,164(1):580-586
The major components of the mitosis-specific histone H1 kinase are CDC2 kinase and cyclin and the consensus amino acid sequence for phosphorylation by this enzyme has been proposed. We have noted the presence of such sequences in six sites of the tumor suppressor gene RB protein and determined whether or not RB protein is in fact phosphorylated by this kinase. Highly purified enzyme was used for this purpose. HeLa cell extracts immunoprecipitated with anti-RB antiserum as well as RB proteins expressed in E. coli cells were shown to be phosphorylated by this kinase in vitro. Synthetic peptides for the six expected sites were also phosphorylated. These results suggest the possibility that the function of RB protein is regulated by CDC2 kinase. 相似文献
22.
Masahito Tsuboi Jun Shoji Atsushi Sogabe Ingrid Ahnesjö Niclas Kolm 《Ecology and evolution》2016,6(3):647-655
The brain is one of the most energetically expensive organs in the vertebrate body. Consequently, the high cost of brain development and maintenance is predicted to constrain adaptive brain size evolution (the expensive tissue hypothesis, ETH). Here, we test the ETH in a teleost fish with predominant female mating competition (reversed sex roles) and male pregnancy, the pacific seaweed pipefish Syngnathus schlegeli. The relative size of the brain and other energetically expensive organs (kidney, liver, heart, gut, visceral fat, and ovary/testis) was compared among three groups: pregnant males, nonpregnant males and egg producing females. Brood size in pregnant males was unrelated to brain size or the size of any other organ, whereas positive relationships were found between ovary size, kidney size, and liver size in females. Moreover, we found that the size of energetically expensive organs (brain, heart, gut, kidney, and liver) as well as the amount of visceral fat did not differ between pregnant and nonpregnant males. However, we found marked differences in relative size of the expensive organs between sexes. Females had larger liver and kidney than males, whereas males stored more visceral fat than females. Furthermore, in females we found a negative correlation between brain size and the amount of visceral fat, whereas in males, a positive trend between brain size and both liver and heart size was found. These results suggest that, while the majority of variation in the size of various expensive organs in this species likely reflects that individuals in good condition can afford to allocate resources to several organs, the cost of the expensive brain was visible in the visceral fat content of females, possibly due to the high costs associated with female egg production. 相似文献
23.
Yuka Okada Yuko Ueshin Hidetoshi Hasuwa Kazuhiro Takumi Masaru Okabe Masahito Ikawa 《Genesis (New York, N.Y. : 2000)》2009,47(4):217-223
Lentiviral vectors efficiently integrate into the host genome of both dividing and nondividing cells, and so they have been used for stable transgene expression in biological and biomedical studies. However, recent studies have highlighted the risk of insertional mutagenesis and subsequent oncogenesis. Here, we used an integrase‐defective lentiviral (IDLV) vector to decrease the chance of random integration and examined the feasibility of lentiviral vector‐mediated gene targeting into murine embryonic stem (ES) cells. After transduction with wild‐type lentiviral vectors, none of the 512 G418 resistant clones were found to be homologous recombinant clones. Although the transduction efficiency was lower with the IDLV vectors (5.9% of wild‐type), successful homologous recombination was observed in nine out of the 941 G418 resistant clones (0.83 ± 1.32%). Pluripotency of the homologous recombinant ES cells was confirmed by the production of chimeric mice and subsequent germ line transmission. Because lentiviral vectors can efficiently transduce a variety of stem cell types, our strategy has potential relevance for secure gene‐manipulation in therapeutic applications. genesis 47:217–223, 2009. © 2009 Wiley‐Liss, Inc. 相似文献
24.
Real-time detection of DNA hybridization on microarray using a CCD-based imaging system equipped with a rotated microlens array disk 总被引:2,自引:0,他引:2
Mogi T Hatakeyama K Taguchi T Wake H Tanaami T Hosokawa M Tanaka T Matsunaga T 《Biosensors & bioelectronics》2011,26(5):1942-1946
This work describes a novel charge-coupled device (CCD)-based imaging system (MB Biochip Reader?) for real-time detection of DNA hybridization to DNA microarrays. The MB Biochip Reader? consisted of a laser light source (532 nm), a microlens array for generation of a multi-beam laser, and a CCD for 2-D signal imaging. The MB Biochip Reader? with a rotated microlens array, allowed large-field imaging (6.2 mm × 7.6 mm with 6.45 μm resolution) with fast time-resolution at 0.2 s without speckle noise. Furthermore, real-time detection of DNA hybridization, which is sufficient to obtain accurate data from tens of thousands of array element per field, was successfully performed without the need for laser scanning. The performance of the MB Biochip Reader? for DNA microarray imaging was similar to the commercially available photomultiplier tube (PMT)-based microarray scanner, ScanArray Lite. The system potentially could be applied toward real-time analysis in many other fluorescent techniques in addition to real-time DNA microarray analysis. 相似文献
25.
目的检测γ-氨基丁酸(gamma-aminobutyric acid,GABA)和谷氨酸脱羧酶(glutamic acid decarboxylase,GAD)在大鼠降结肠上皮的表达及分布特征,并探讨GABA与上皮细胞分化增殖的关系。方法用免疫荧光及激光共聚焦显微扫描技术,检测GABA、GAD65及GAD67在大鼠降结肠上皮中的表达,并以麦芽凝聚素组织化学染色与免疫荧光结合的双重染色显示GABA和GAD65表达细胞的分布特征。同时,用RT-PCR方法检测GAD mRNA的表达。此外,用3H-胸腺嘧啶放射自显影及增殖细胞核抗原(PCNA)免疫组化方法显示降结肠上皮的增殖带。结果RT-PCR显示降结肠粘膜中GAD65及GAD67mRNA均为阳性。GABA及GAD65免疫反应阳性细胞主要分布在降结肠的腔面和隐窝的上1/3上皮细胞的胞浆,而GAD67阳性细胞仅分布腔面,此外,GABA及GAD65阳性染色也见于黏膜固有层。双重染色显示杯状细胞中GABA及GAD65均为阴性3。H-胸腺嘧啶及PCNA标记阳性细胞主要在隐窝的中下段。结论GABA及GAD65分布在大鼠降结肠上皮的成熟带及功能带,GABA系统可能参与上皮细胞的分化与增殖的调节。 相似文献
26.
The peroxisomal core from the liver of rats was purified 450-fold as a marker of urate oxidase [EC 1.7.3.3.] activity. This preparation has a high specific activity of urate oxidase but not of other peroxisomal enzymes: D-amino acid oxidase [EC 1.4.3.3.], L-alpha-hydroxy acid oxidase [EC 1.1.3.15], or catalase [EC 1.11.1.6]. No activity of marker enzymes for other subcellular particles; cytochrome c oxidase [EC1.9.3.1] (mitochondria), acid phosphatase [EC 3.1.3.2] (lysosomes), or glucose-6-phosphatase [EC 3.1.3.9] (microsomes), was detected in this preparation. The core obtained showed a single protein band in sodium dodecyl sulfate-polyacrylamide gel electrophoresis and the position of the band was found to correspond to a molecular weight 35,000. When the peroxisomal core was subjected to treatment at various pH's with 0.1 M carbonate buffer, urate oxidase was almost completely solubulized at pH 11.0, although approximately 35% of the core protein still remained in the pellet After solubilization of the core at pH 11.0, the specific activity of urate oxidase in the supernatant increased about 1.6 times; the density of the insoluble protein remaining in the pellet was identical with the that of the original core on sucrose density gradient centrifugation. 相似文献
27.
Go Suzuki Wataru Okada Yoko Yasutake Hidekazu Yamamoto Iwao Tanita Hiroshi Yamashita Takeshi Hayashibara Toshiaki Komatsu Toru Kanyama Masahito Inoue Masashi Yamazaki 《Restoration Ecology》2020,28(5):1172-1182
Larval recruitment is essential for sustaining coral communities and a fundamental tool in some interventions for reef restoration. To improve larval supply and post‐settlement survival in sexually assisted coral restoration efforts, an integrated in situ collector system, the larval cradle, was designed to collect spawned gametes then culture the resulting larvae until settled on artificial substrates. The final design of the larval cradle was cylindrical, a nylon mesh structure with a volume of 9 m3, suspended in the sea and extending vertically toward the seabed. We found three key design features that improved the efficiency of the apparatus: (1) an open area of sea surface and mesh size of less than 100 μm produced high fertilization and optimal survival (>90%), (2) a special skirt‐shaped net (3 m in diameter) with a connection hose for attaching the cradle to collect bundles from many adult colonies over a wide area and at various depths, and (3) adding short square tube pieces, called square hollow sections, as a substrate for enhancing larval settlement and survival, to a larval cradle at 4 days after spawning was optimal for uniform settlement. This system allowed not only the collection of several million eggs, but also subsequent production of several thousand settled juvenile corals, without land facilities. Our design achieved several hundred times higher survival for early life stages of Acropora tenuis compared to nature. 相似文献
28.
Zhihua Liu John S. Kimball Nicholas C. Parazoo Ashley P. Ballantyne Wen J. Wang Nima Madani Caleb G. Pan Jennifer D. Watts Rolf H. Reichle Oliver Sonnentag Philip Marsh Miriam Hurkuck Manuel Helbig William L. Quinton Donatella Zona Masahito Ueyama Hideki Kobayashi Eugnie S. Euskirchen 《Global Change Biology》2020,26(2):682-696
Arctic and boreal ecosystems play an important role in the global carbon (C) budget, and whether they act as a future net C sink or source depends on climate and environmental change. Here, we used complementary in situ measurements, model simulations, and satellite observations to investigate the net carbon dioxide (CO2) seasonal cycle and its climatic and environmental controls across Alaska and northwestern Canada during the anomalously warm winter to spring conditions of 2015 and 2016 (relative to 2010–2014). In the warm spring, we found that photosynthesis was enhanced more than respiration, leading to greater CO2 uptake. However, photosynthetic enhancement from spring warming was partially offset by greater ecosystem respiration during the preceding anomalously warm winter, resulting in nearly neutral effects on the annual net CO2 balance. Eddy covariance CO2 flux measurements showed that air temperature has a primary influence on net CO2 exchange in winter and spring, while soil moisture has a primary control on net CO2 exchange in the fall. The net CO2 exchange was generally more moisture limited in the boreal region than in the Arctic tundra. Our analysis indicates complex seasonal interactions of underlying C cycle processes in response to changing climate and hydrology that may not manifest in changes in net annual CO2 exchange. Therefore, a better understanding of the seasonal response of C cycle processes may provide important insights for predicting future carbon–climate feedbacks and their consequences on atmospheric CO2 dynamics in the northern high latitudes. 相似文献
29.
30.
Masrina Mohd Nadzir Masahiro Kino-oka Katsura Sugawara Masahito Taya 《Biotechnology letters》2013,35(2):295-300
The effect of insulin-like growth factor-1 (IGF-1) on the behavior of rabbit chondrocytes in cultured collagen (CL) gels initially seeded with 2 × 105 cells/ml was examined. On day 5, the frequency of migrating cells cultured in presence of 100 ng IGF-1/ml was 0.04, which was 54 % of the frequency in IGF-1-free culture. The presence of IGF-1 caused an increase in the frequency of dividing cells from 0.09 to 0.13. These results suggest that IGF-1 suppressed the migration of chondrocytes in the CL gels while stimulating cell division in the initial culture phase. The proteolytic migration of cells was thought to be suppressed by the down-regulation of membrane type 1 matrix metalloproteinase by IGF-1. This contributed to the formation of aggregates with spherical-shaped cells that produced collagen type II. 相似文献