全文获取类型
收费全文 | 424篇 |
免费 | 47篇 |
国内免费 | 14篇 |
出版年
2024年 | 2篇 |
2023年 | 8篇 |
2022年 | 12篇 |
2021年 | 17篇 |
2020年 | 20篇 |
2019年 | 19篇 |
2018年 | 20篇 |
2017年 | 18篇 |
2016年 | 7篇 |
2015年 | 15篇 |
2014年 | 45篇 |
2013年 | 24篇 |
2012年 | 18篇 |
2011年 | 27篇 |
2010年 | 7篇 |
2009年 | 9篇 |
2008年 | 11篇 |
2007年 | 22篇 |
2006年 | 11篇 |
2005年 | 13篇 |
2004年 | 10篇 |
2003年 | 9篇 |
2002年 | 6篇 |
2001年 | 8篇 |
2000年 | 5篇 |
1999年 | 10篇 |
1998年 | 5篇 |
1997年 | 12篇 |
1996年 | 10篇 |
1995年 | 12篇 |
1994年 | 17篇 |
1993年 | 8篇 |
1992年 | 4篇 |
1991年 | 6篇 |
1990年 | 6篇 |
1989年 | 2篇 |
1988年 | 4篇 |
1987年 | 6篇 |
1985年 | 3篇 |
1984年 | 2篇 |
1983年 | 2篇 |
1982年 | 2篇 |
1981年 | 1篇 |
1980年 | 1篇 |
1979年 | 2篇 |
1978年 | 1篇 |
1976年 | 2篇 |
1974年 | 1篇 |
1971年 | 1篇 |
1970年 | 1篇 |
排序方式: 共有485条查询结果,搜索用时 62 毫秒
1.
《European journal of cell biology》2022,101(3):151256
An in vitro bone triple culture involving human primary osteoblasts, osteocytes and osteoclasts enables the investigation of bone healing factors, drugs or biomaterials in a model system for native bone tissue. The present study analyses the impact of Sr2+ as well as hypoxic cultivation (5% O2 content or chemically induced by Co2+) on bone cells. The three cell types were cultivated together in the presence of 100 µM Sr2+, hypoxic conditions or in the presence of 75 µM Co2+. After cultivation the cell types were separated and analysed on mRNA and protein level individually. In response to Sr2+ osteoblasts showed a downregulation of IBSP expression and a stimulation of ALP activity. Osteocyte gene marker expression of PDPN, MEPE, RANKL, OPG, osteocalcin and likewise the amount of secreted osteocalcin was reduced in the presence of Sr2+. Activity of osteoclast-specific enzymes TRAP and CAII was enhanced compared to the Sr2+ free control. Hypoxic conditions induced by both 5% O2 or a Co2+ treatment led to decreased DNA content of all bone cells and downregulated expression of osteoblast markers ALPL and IBSP as well as osteocyte markers PDPN, RANKL and OPG. In addition, Co2+ induced hypoxia decreased gene and protein expression of osteocalcin in osteocytes. In response to the Co2+ treatment, the TRAP gene expression and activity was increased. This study is the first to analyse the effects of Sr2+ or hypoxia on triple cultures with primary human bone cells. The investigated in vitro bone model might be suitable to reduce animal experiments in early stages of biomaterial and drug development. 相似文献
2.
The capsular polysaccharide (CPS) of Staphylococcus aureus strain Smith was labelled by growth of bacteria in the presence of radioactive N-acetylglucosamine and was separated from labelled cell wall components by affinity chromatography on wheat germ agglutinin following dissolution of the cells by lysostaphin. The products were partially characterised chemically and immunochemically. Similar labelled components were found in the culture fluid during growth. In a pulse-chase experiment, cell-bound CPS was released continuously into the culture fluid at the same rate as cell wall turnover and there was no evidence of direct excretion of CPS. 相似文献
3.
The intracellular growth of pathogenic mycobacteria has been linked to the presence of an electron transparent zone (ETZ or capsule), which surrounds the phagocytized bacteria and prevents the diffusion of lysosomal enzymes in infected macrophages. Recently, it was suggested that this capsule may be a bacterial structures, even being present in test tube-grown pathogenic mycobacteria (FEMS Microbiol. Lett. 1988, 56, 225-230). In the present paper, we show that under special fixation and embedding conditions, this capsule was clearly observed among 7 strains of mycobacteria grown in axenic media and also in M. leprae extracted and purified from experimentally infected armadillo or nude mice. In the case of bacteria treated likewise but subject to a prior dehydration step, this capsular structure disappeared suggesting its lipidic nature. Ultrathin sections of M. intracellular after immunolabelling showed for the first time that this capsule obtained mycobacterial antigens confirming its mycobacterial origin. It is suggested that the mycobacterial capsule may be formed of inert lipids, in which surface antigens are embedded. 相似文献
4.
Summary A 3D1H–15N–13C triple resonance experiment is presented that contains exclusively cross peaks between the1HN and15N nuclei of one residue with the H of the preceding residue. The pulse sequence, designed to minimize the time coherence, is transverse on nuclei with short T2 values. The experiment consists of coherence transfers via one-bond couplings from the HN via N, CO, C to the H and back to the HN for detection; it is called HN(COCA)HA. The experiment was tested on uniformly15N- and13C-enriched T4 lysozyme. 相似文献
5.
Polymer chains of (13)--d-glucan were dissolved with 1 M NaOH at 4° C from native microfibrillar protoplast nets. The chains associated into microfibrils during NaOH neutralization or dialysis. In contrast to the native microfibrils which are of uniform width individually (10 to 20 nm) and arranged in flat bundles, the microfibrils formed in vitro showed no band formation and consisted of fibrous spindle-shaped subunits of variable width or loose elementary fibrils about 1.7 nm wide. X-ray diagrams of native nets indicated a fairly high crystallinity and were different for wet and dry specimens. They corresponded to those of paramylon. Precipitated glucans produced diagrams different from the former and revealing a lower crystallinity especially with the dry samples.The X-ray pattern, combined with other data, allowed the precipitated microfibrils to be identified as aggregates of molecular strands composed each of three intertwined helical glucan chains. Since these triple helical chains are about 1.7 nm wide the elementary fibrils of this width can represent only single triple-helical strands. These helices have 7 glucose residues per turn and therefore a low symmetry which explains the poor crystallizing properties. The 7 membered helix represents a basic difference with the well crystallized native glucan which is built of highly symmetrical triple helices with 6 glucose residues per turn. Since 61 helical conformation is not formed in vitro at normal temperatures its generation in vivo must be due to the action of synthesizing enzymes at the protoplast membrane. The intertwining of these helices and crystallization of the strands are determined by their symmetry and physical properties of the chains. This characterizes the native microfibrils as products of self-assembly of enzymegenerated 61 helices. 相似文献
6.
A. M. Junelles A. El Kanouni H. Petitdemange R. Gay 《Journal of industrial microbiology & biotechnology》1989,4(2):121-125
Summary The production of granulose (an intracellular reserve polygranule), capsule and exopolysaccharide was investigated in a synthetic medium in which the oxido-reduction level was modified by the addition of acetic or butyric acid. After addition of the acids, granulose synthesis increased from 150 to 300 mg glucose equivalents ·1–1 and capsular synthesis decreased by 25%. Exopolysaccharide production was unchanged under these conditions. A hypothesis that attributes a role to the polymer in the oxido-reduction sequences is discussed. 相似文献
7.
在灭菌自来水模拟水体中,研究了7种细菌的存活和生长规律。Klebsiella pneumo-niae,Enterobacter aerogenes,Agrobacterium tumefatciens,在7天内平板计数降至0,而水体中镜检细菌总数(AODC)和活菌直接计数(DVC)结果无大变化,说明细菌已变成活的非可培养状态。Micrococcus,flavus 和 Streptococcus faecalis 的可培养菌数也可降至0。Pseudomonas sp.在48小时内由10~5降至10~2cfu/ml,随即升至10~6 cfu/ml 并持续到实验终了(41天)。Bacillus subtilis 在48小时平板计数降至10~2cfu/ml 并维持在该水平至实验结束(38天)。研究结果表明仅用涂布平板法检测多种细菌在水环境中的生存和分布是不合适的。 相似文献
8.
Resuscitation of Vibrio cholerae O1 strain TSI-4 from a viable but nonculturable state by heat shock 总被引:3,自引:0,他引:3
Sun Nyunt Wai Tetsuhiro Moriya Katsuhiko Kondo Hiroyasu Misumi Kazunobu Amako 《FEMS microbiology letters》1996,136(2):187-191
Abstract Vibrio cholerae strain TSI-4 was incubated in an M9 salt solution at 15 °C for more than 100 days. The plate counts showed no viable cells on day 30, but a broth culture from that day showed the growth of bacteria. However, after 35 days the bacteria entered the nonculturable state, based on the assessment of both the plate counts and broth culture. A portion of the culture was heated at 45 °C for 1 min in a water bath and subsequently plated onto a nutrient agar plate. More than 1000 colonies were recovered after this heat-shock treatment. The recovered cells showed the same chromosomal DNA pattern in the restriction map and the same outer membrane protein pattern in SDS-PAGE. Recovery of viable cells by heat-shock was achieved in cultures grown on M9 salt but not from cultures grown in phosphate-buffered saline. This suggests that the presence of NH4 Cl in the M9 salt solution may support the growth of the bacteria in a low nutrient medium, while also playing an important role in resuscitation. 相似文献
9.
大豆灰斑病抗性遗传的三点测交分析 总被引:1,自引:0,他引:1
本实验利用三点测交分析的方法, 对3个组合在人工接种大豆灰斑病菌的条件下的抗性表现进行基因效应分析,各组合均存在加性,组合1存在显性,组合2、3存在上位性。
Abstract:In this paper,Triple Test Cross Design was used in studing the resistance of soybean to 10 physiological race of Cercospora Sojina Haraby inoculation.Results o analysis of gene effects of resistance indicated that additive effect is significant in all the three crosses,dominant effect exsists only in the cross 1 and epistatic effect remains in the cross 2 and cross 3. 相似文献
10.
Olivier Bornet Gérard Lancelot Luc Chanteloup Nguyen T. Thuong Jean-Marie Beau 《Journal of biomolecular NMR》1994,4(4):575-580
Summary We present NMR studies of an intramolecular triple helix, the three strands of which have been linked by a hexaethylene glycol chain. To overcome the generally encountered difficulties of assignment in the homopyrimidine strands, the carbon C1 of the pyrimidines were selectively 13C-enriched. Assignments of the aromatic and sugar protons were obtained from NOESY-HMQC and TOCSY-HMQC spectra. We show that the recognition of a DNA duplex by a third strand via triplex formation is easily carried out in solution by observing the changes of the 1H1–13C1 connectivities as a function of pH. Furthermore, the conformation of the sugars has been found to be C2-endo, on the basis of the coupling constant values directly measured in an HSQC spectrum. 相似文献