全文获取类型
收费全文 | 2336篇 |
免费 | 117篇 |
国内免费 | 1篇 |
出版年
2021年 | 30篇 |
2020年 | 18篇 |
2019年 | 19篇 |
2018年 | 16篇 |
2017年 | 25篇 |
2016年 | 44篇 |
2015年 | 54篇 |
2014年 | 78篇 |
2013年 | 188篇 |
2012年 | 133篇 |
2011年 | 141篇 |
2010年 | 82篇 |
2009年 | 65篇 |
2008年 | 133篇 |
2007年 | 138篇 |
2006年 | 130篇 |
2005年 | 152篇 |
2004年 | 122篇 |
2003年 | 127篇 |
2002年 | 154篇 |
2001年 | 33篇 |
2000年 | 21篇 |
1999年 | 30篇 |
1998年 | 49篇 |
1997年 | 26篇 |
1996年 | 24篇 |
1995年 | 17篇 |
1994年 | 32篇 |
1993年 | 20篇 |
1992年 | 19篇 |
1991年 | 20篇 |
1990年 | 20篇 |
1989年 | 25篇 |
1988年 | 24篇 |
1987年 | 20篇 |
1986年 | 18篇 |
1985年 | 12篇 |
1984年 | 20篇 |
1983年 | 21篇 |
1982年 | 14篇 |
1981年 | 21篇 |
1980年 | 19篇 |
1979年 | 10篇 |
1978年 | 7篇 |
1977年 | 7篇 |
1976年 | 10篇 |
1975年 | 10篇 |
1974年 | 10篇 |
1972年 | 6篇 |
1971年 | 8篇 |
排序方式: 共有2454条查询结果,搜索用时 296 毫秒
1.
Kei Watanabe Kenta Wada Tomoko Ohashi Saki Okubo Kensuke Takekuma Ryoichi Hashizume Jun-Ichi Hayashi Tadao Serikawa Takashi Kuramoto Yoshiaki Kikkawa 《PloS one》2012,7(11)
We discovered a new cataract mutation, kfrs4, in the Kyoto Fancy Rat Stock (KFRS) background. Within 1 month of birth, all kfrs4/kfrs4 homozygotes developed cataracts, with severe opacity in the nuclei of the lens. In contrast, no opacity was observed in the kfrs4/+ heterozygotes. We continued to observe these rats until they reached 1 year of age and found that cataractogenesis did not occur in kfrs4/+ rats. To define the histological defects in the lenses of kfrs4 rats, sections of the eyes of these rats were prepared. Although the lenses of kfrs4/kfrs4 homozygotes showed severely disorganised fibres and vacuolation, the lenses of kfrs4/+ heterozygotes appeared normal and similar to those of wild-type rats. We used positional cloning to identify the kfrs4 mutation. The mutation was mapped to an approximately 9.7-Mb region on chromosome 7, which contains the Mip gene. This gene is responsible for a dominant form of cataract in humans and mice. Sequence analysis of the mutant-derived Mip gene identified a 5-bp insertion. This insertion is predicted to inactivate the MIP protein, as it produces a frameshift that results in the synthesis of 6 novel amino acid residues and a truncated protein that lacks 136 amino acids in the C-terminal region, and no MIP immunoreactivity was observed in the lens fibre cells of kfrs4/kfrs4 homozygous rats using an antibody that recognises the C- and N-terminus of MIP. In addition, the kfrs4/+ heterozygotes showed reduced expression of Mip mRNA and MIP protein and the kfrs4/kfrs4 homozygotes showed no expression in the lens. These results indicate that the kfrs4 mutation conveys a loss-of-function, which leads to functional inactivation though the degradation of Mip mRNA by an mRNA decay mechanism. Therefore, the kfrs4 rat represents the first characterised rat model with a recessive mutation in the Mip gene. 相似文献
2.
3.
Ichiro Tomida Hiroshi Kayahara Ryozo Iriye 《Bioscience, biotechnology, and biochemistry》2013,77(11):2557-2563
The degree of racemization in the coupling reaction, Pro-Val + Pro, with the several other methods than the activated ester methods was measured and the results were compared with that in the coupling reaction, Leu-Phe + Val, as well as in the previous paper.1) In the azide and the mixed anhydride methods, no or almost no racemization was detected, whereas in the other tested methods of peptide synthesis (Pachornik-, DCCD- and phosphazo-methods) the significantly large racemization was observed. It can be attributed to the strong nucleo- philic N-atom in the penultimate amino acids (Pro) and the steric hindrance of C-terminal amino acid (Val), which are favourable to the formation of the oxazolone ring.This assumption was further systematically confirmed in the synthesis of the other several tripeptides with the DCCD method. The separation of the diastereoisomers (LLL and LDL) of the resulting tripeptides by gas chromatography with a packed column was also here reported. 相似文献
4.
The isolation and culture of lily pollen protoplasts 总被引:4,自引:0,他引:4
Methods for the enzymatic isolation of lily protoplasts and their successful culture are described. When pre-anthesis binucleate pollen (immature pollen grains) was treated in enzyme solution containing macerozyme and cellulase, up to 80% lost their exine and gave rise to intact protoplasts within 1 h. These pollen protoplasts were uniform in size and densely cytoplasmic with two prominent generative and vegetative nuclei. The isolated pollen protoplasts regenerated a cell wall within 1 day of culture and produced a structure resembling a pollen tube after 10–12 days of culture. During this culture period, dividing generative nuclei or 2 sperm nuclei were observed in many protoplasts with regenerated cell walls. 相似文献
5.
Toshifumi Nakao Ichiro Yamato Yasuhiro Anraku 《Molecular & general genetics : MGG》1987,208(1-2):70-75
6.
Differential expression of multiple protein kinase C subspecies in rat central nervous tissue 总被引:16,自引:0,他引:16
M S Shearman Z Naor U Kikkawa Y Nishizuka 《Biochemical and biophysical research communications》1987,147(3):911-919
Protein kinase C from a number of areas of rat central nervous tissue was resolved into three distinct fractions upon hydroxyapatite column chromatography. One of the enzyme fractions, designated type II, could be further distinguished into two subspecies with polyclonal antisera, which were raised against synthetic peptides specific for the predicted amino acid sequences of two alternative cDNA clones encoding this enzyme type. Using a combination of these biochemical and immunological techniques, the relative activity of the multiple subspecies of protein kinase C was assessed for each brain area. A distinct regional pattern of expression was found, which per se may be an important factor in determining the response of different neuronal cell types to extracellular stimuli. 相似文献
7.
Three distinct forms of rat brain protein kinase C: differential response to unsaturated fatty acids 总被引:18,自引:0,他引:18
K Sekiguchi M Tsukuda K Ogita U Kikkawa Y Nishizuka 《Biochemical and biophysical research communications》1987,145(2):797-802
Although the three distinct forms of protein kinase C isolated from rat brain soluble fraction are structurally very similar, they respond differently to free unsaturated fatty acids such as arachidonic acid to exhibit their catalytic activity. Type I enzyme encoded by gamma-sequence, as predicted by cDNA clone analysis, responds to these fatty acids only slightly, whereas Type III enzyme determined by alpha-sequence is activated by free unsaturated fatty acids in the presence of Ca2+ in a comparable manner to phosphatidylserine plus diacylglycerol. Type II, a mixture of two enzymes encoded by beta I- and beta II-sequence, resulting from alternative splicing, shows properties in between those of Type I and Type III. Some of these forms of protein kinase C may function at a relatively later phase of cellular responses when large amounts of unsaturated fatty acids and Ca2+ are mobilized. 相似文献
8.
Mikihiko Naito Ichiro Kudo Yukiko Mukai-Sato Susumu Tsushima Hiroaki Nomura Shoshichi Nojima Keizo Inoue 《Cancer immunology, immunotherapy : CII》1987,24(2):158-164
Summary Liposomes composed of chemically synthesized glyceroglycolipids, such as 1,2-dipalmityl-[-cellobiosyl-(1 3)]-glycerol (Cel-DAG), 1,2-dipalmityl-[-lactosyl-(1 3)]-glycerol, or 1,2-dipalmityl-[-maltosyl-(1 3)]-glycerol, were found to enhance protective immunity against transplantable tumor cells (sarcoma 180) in ICR mice. Peritoneal exudate cells prepared from mice treated in vivo with Cel-DAG showed cytostatic activity in vitro against the mouse leukemia cell line, EL-4. Adherent cells separated from this preparation showed similar activity. Peritoneal cells from polypeptone-injected mice acquired appreciable cytostatic activity when incubated in vitro in the presence of glyceroglycolipid liposomes. The adherent cell fraction alone showed rather weak cytostatic activity when pretreated with the glyceroglycolipids, and full activity was restored by supplementing with the nonadherent cell fraction. The ability of glycolipids to induce tumoricidal effects was affected by cholesterol content: with increasing cholesterol content, the activities decreased. Cholesterol-free glycolipid liposomes were taken more efficiently by macrophages than cholesterol-containing liposomes. Cholersterol modifies the surface property of glyceroglycolipid liposomes. Activation of macrophages is responsible for enhancement of protective immunity against tumor cells by injection of these glycolipids in vivo.This work was supported in part by Grants-in-Aid (Nos. 58010010, and 59870076) for Scientific Research from the Ministry of Education, Science and Culture of Japan 相似文献
9.
Obi Ichiro; Ichikawa Yoshiaki; Kakutani Tadaaki; Senda Mitsugi 《Plant & cell physiology》1989,30(3):439-444
Electrophoretic mobilities of plant protoplasts from varioussources were measured, as a function of the pH of the medium,by a micro-electrophoresis technique to characterize the protoplastsin terms of curves of zeta potential vs. protoplast surfacepH (pHs). The shape of the curves of zeta potential vs. pHscurves differed among preparations of protoplasts isolated fromvarious species and strains. The isoelectric points (pI) ofthe protoplasts measured in this study were between 3.0 and4.0. These differences among the protoplasts suggest that itmay be possible to develop an electrophoretic method for theselection of protoplasts. The shape of the curves of zeta potentialvs. pHs also indicated that carboxyl groups, as well as phosphategroups, may contribute to the negative charges on the surfaceof protoplasts. (Received October 14, 1988; Accepted February 22, 1989) 相似文献
10.
Time-resolved x-ray diffraction and calorimetric studies at low scan rates: I. Fully hydrated dipalmitoylphosphatidylcholine (DPPC) and DPPC/water/ethanol phases 总被引:4,自引:4,他引:0 下载免费PDF全文
The phase transitions in fully hydrated dipalmitoylphosphatidylcholine (DPPC) and DPPC/water/ethanol phases have been studied by lowangle time-resolved x-ray diffraction under conditions similar to those employed in calorimetry (scan rates 0.05-0.5°C/min and uniform temperature throughout the samples). This approach provides more adequate characterization of the equilibrium transition pathways and allows for close correlations between structural and thermodynamic data. No coexistence of the rippled gel (Pβ') and liquid-crystalline (Lα) phases was found in the main transition of DPPC; rather, a loss of correlation in the lamellar structure, observed as broadening of the lamellar reflections, takes place in a narrow temperature range of ~100 mK at the transition midpoint. Formation of a long-living metastable phase, denoted by Pβ'(mst), differing from the initial Pβ' was observed in cooling direction by both x-ray diffraction and calorimetry. No direct conversion of Pβ'(mst) into Pβ' occurs for over 24 h but only by way of the phase sequence Pβ'(mst) → Lβ' → Pβ'. According to differential scanning calorimetry (DSC), the enthalpy of the Pβ'(mst)-Lα transition is by ~5% lower than that of the Pβ'-Lα transition. The effects of ethanol (Rowe, E. S. 1983. Biochemistry. 22:3299-3305; Simon, S. A., and T. J. McIntosh. 1984. Biochim. Biophys. Acta 773:169-172) on the mechanism and reversibility of the DPPC main transition were clearly visualized. At ethanol concentrations inducing formation of interdigitated gel phase, the main transition proceeds through a coexistence of the initial and final phases over a finite temperature range. During the subtransition in DPPC recorded at scan rate 0.3°C/min, a smooth monotonic increase of the lamellar spacing from its subgel (Lc) to its gel (Lβ') phase value takes place. The width of the lamellar reflections remains unchanged during this transformation. This provides grounds to propose a “sequential” relaxation mechanism for the subgel-gel transition which is not accompanied by growth of domains of the final phase within the initial one. 相似文献