全文获取类型
收费全文 | 2593篇 |
免费 | 223篇 |
国内免费 | 1篇 |
出版年
2023年 | 8篇 |
2022年 | 31篇 |
2021年 | 42篇 |
2020年 | 47篇 |
2019年 | 64篇 |
2018年 | 59篇 |
2017年 | 53篇 |
2016年 | 72篇 |
2015年 | 113篇 |
2014年 | 147篇 |
2013年 | 185篇 |
2012年 | 228篇 |
2011年 | 243篇 |
2010年 | 132篇 |
2009年 | 88篇 |
2008年 | 138篇 |
2007年 | 149篇 |
2006年 | 146篇 |
2005年 | 112篇 |
2004年 | 149篇 |
2003年 | 134篇 |
2002年 | 110篇 |
2001年 | 46篇 |
2000年 | 35篇 |
1999年 | 38篇 |
1998年 | 22篇 |
1997年 | 20篇 |
1996年 | 19篇 |
1995年 | 12篇 |
1994年 | 11篇 |
1993年 | 9篇 |
1992年 | 24篇 |
1991年 | 12篇 |
1990年 | 9篇 |
1989年 | 6篇 |
1988年 | 9篇 |
1987年 | 8篇 |
1986年 | 15篇 |
1985年 | 11篇 |
1984年 | 5篇 |
1983年 | 8篇 |
1982年 | 6篇 |
1980年 | 4篇 |
1979年 | 4篇 |
1977年 | 2篇 |
1976年 | 3篇 |
1974年 | 3篇 |
1973年 | 2篇 |
1970年 | 3篇 |
1964年 | 2篇 |
排序方式: 共有2817条查询结果,搜索用时 46 毫秒
1.
M. Graciela Pucciarelli S. Ruschkowski T.J. Trust B.B. Finlay 《FEMS microbiology letters》1995,129(2-3):293-399
Abstract Helicobacter pylori is a bacterial pathogen of humans that infects the gastric mucosa. This infection has been associated with gastritis, peptic ulcers, and gastric carcinomas. Diverse in vitro studies have described efficient adherence of H. pylori to different types of epithelial cells. Because of its varied effects on host cells, we have analysed signal transduction events in H. pyfori -infected epithelial cells. Our results show that H. pylori induces an increase in inositol phosphates in all cultured epithelial cells used, including HeLa, Henle 407, Hep-2, and the human gastric adenocarcinoma cell line AGS. Bacterial growth medium supernatants induce a similar response in the host cell. The increase in inositol phosphates is not related to redistribution of cytoskeletal proteins such as actin or α-actinin nor tyrosine-phosphorylation of host cell proteins. The inositol phosphate increase is also observed in cells infected with low or non-adherent H. pylori mutants or mutants defective in the vacuolating toxin or urease holoenzyme. These results indicate that inositol phosphate release in H. pytori -infected cells is not dependent on bacterial adherence, and that a soluble bacterial factor, but not the vacuolating toxin or urease holoenzyme, mediates such an effect. 相似文献
2.
Abstract. 1. Pollen is considered to be an important dietary component for many species of flower-feeding herbivores. Its influence on oviposition site selection by the pollen beetle Meligethes aeneus , and on the development of its larvae was investigated.
2. The effects of pollen presence and absence on adult, egg, and larval incidence in the field, and on larval development in the laboratory were compared through the use of Synergy, a composite hybrid oilseed rape Brassica napus variety comprising male-fertile (with pollen) and male-sterile (without pollen) plants.
3. In the field, adult females were more abundant on male-fertile plants during flowering, and a greater proportion of male-fertile than male-sterile buds were accepted for oviposition. These data indicate a possible role of pollen in oviposition site selection by female pollen beetles.
4. The numbers of first instar larvae on the two plant lines did not differ; however, more second instars were found on male-fertile than on male-sterile flowers. This suggests a greater larval survival on male-fertile plants, possibly due to the more readily available food resources and better nutrition afforded by the presence of pollen.
5. Laboratory experiments confirmed that a diet which included pollen improved survival to adulthood and resulted in heavier pupae and adults; however, pollen was not obligatory for larval survival and development.
6. The pollen beetle, previously thought to be an obligate pollen feeder, is therefore more generalist in its requirements for development. These findings may relate to the nutritional and behavioural ecology of other flower-feeding herbivores. 相似文献
2. The effects of pollen presence and absence on adult, egg, and larval incidence in the field, and on larval development in the laboratory were compared through the use of Synergy, a composite hybrid oilseed rape Brassica napus variety comprising male-fertile (with pollen) and male-sterile (without pollen) plants.
3. In the field, adult females were more abundant on male-fertile plants during flowering, and a greater proportion of male-fertile than male-sterile buds were accepted for oviposition. These data indicate a possible role of pollen in oviposition site selection by female pollen beetles.
4. The numbers of first instar larvae on the two plant lines did not differ; however, more second instars were found on male-fertile than on male-sterile flowers. This suggests a greater larval survival on male-fertile plants, possibly due to the more readily available food resources and better nutrition afforded by the presence of pollen.
5. Laboratory experiments confirmed that a diet which included pollen improved survival to adulthood and resulted in heavier pupae and adults; however, pollen was not obligatory for larval survival and development.
6. The pollen beetle, previously thought to be an obligate pollen feeder, is therefore more generalist in its requirements for development. These findings may relate to the nutritional and behavioural ecology of other flower-feeding herbivores. 相似文献
3.
John R. Lawrence Pascal J. Delaquis Darren R. Korber Douglas E. Caldwell 《Microbial ecology》1987,14(1):1-14
Phase, darkfield, and computer-enhanced microscopy were used to observe the surface microenvironment of flow cells during bacterial colonization. Microbial behavior was consistent with the assumptions used previously to derive surface colonization kinetics and to calculate surface growth and attachment rates from cell number and distribution. Surface microcolonies consisted of closely packed cells. Each colony contained 2n cells, where n is the number of cell divisions following attachment. Initially, cells were freely motile while attached, performing circular looping movements within the plane of the solid-liquid interface. Subsequently, cells attached apically, maintained a fixed position on the surface, and rotated. This type of attachment was reversible and did not necessarily lead to the formation of microcolonies. Cells became irreversibly attached by progressing from apical to longitudinal attachment. Longitudinally attached cells increased in length, then divided, separated, moved apart laterally, and slid next to one another. This resulted in tight cell packing and permitted simultaneous growth and adherence. After approximately 4 generations, individual cells emigrated from developing microcolonies to recolonize the surface at new locations. Surface colonization byPseudomonas fluorescens can thus be subdivided into the following sequential colonization phases: motile attachment phase, reversible attachment phase, irreversible attachment phase, growth phase, and recolonization phase. 相似文献
4.
Inhibition of in vitro nuclear transport by a lectin that binds to nuclear pores 总被引:88,自引:42,他引:46
下载免费PDF全文
![点击此处可从《The Journal of cell biology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
Selective transport of proteins is a major mechanism by which biochemical differences are maintained between the cytoplasm and nucleus. To begin to investigate the molecular mechanism of nuclear transport, we used an in vitro transport system composed of a Xenopus egg extract, rat liver nuclei, and a fluorescently labeled nuclear protein, nucleoplasmin. With this system, we screened for inhibitors of transport. We found that the lectin, wheat germ agglutinin (WGA), completely inhibits the nuclear transport of fluorescently labeled nucleoplasmin. No other lectin tested affected nuclear transport. The inhibition by WGA was not seen when N-acetylglucosamine was present and was reversible by subsequent addition of sugar. When rat liver nuclei that had been incubated with ferritin-labeled WGA were examined by electron microscopy, multiple molecules of WGA were found bound to the cytoplasmic face of each nuclear pore. Gel electrophoresis and nitrocellulose transfer identified one major and several minor nuclear protein bands as binding 125I-labeled WGA. The most abundant protein of these, a 63-65-kD glycoprotein, is a candidate for the inhibitory site of action of WGA on nuclear protein transport. WGA is the first identified inhibitor of nuclear protein transport and interacts directly with the nuclear pore. 相似文献
5.
Strains of Escherichia, Salmonella, Shigella and Yersinia actively enter eukaryotic cells. Several techniques were used to compare and contrast the invasion mechanisms of Salmonella cholerae-suis, Yersinia enterocolitica and Shigella flexneri. Three animal cell lines (CHO, HEp-2 and MDCK) were examined for susceptibility to bacterial entry by these strains. Levels of intracellular bacteria varied widely between cell lines, but CHO cells were the most susceptible to bacterial invasion, HEp-2 invasion levels were intermediary, whereas polarized MDCK cells were invaded to a lesser extent. This illustrates that tissue culture models can be optimized to study bacterial invasion and intracellular replication. We used these tissue culture models to examine the interactions between host cells and these invasive bacteria. The use of lysosomotropic agents (methylamine and ammonium chloride), cationic ionophores (monensin) and acidification-defective CHO cell lines demonstrated that endosome acidification is not required for bacterial invasion or intracellular replication. Drugs which inhibited microfilament formation (cytochalasins B and D) prevented internalization of S. cholerae-suis, Y. enterocolitica and S. flexneri, indicating that invasion is a microfilament-dependent event. The microtubule inhibitors, colchicine, vincristine and vinblastine, did not affect bacterial internalization. 相似文献
6.
L Svensson B B Finlay D Bass C H von Bonsdorff H B Greenberg 《Journal of virology》1991,65(8):4190-4197
When rotavirus infects the mature villus tip cells of the small intestine, it encounters a highly polarized epithelium. In order to understand this virus-cell interaction more completely, we utilized a cell culture-adapted rhesus rotavirus (RRV) to infect human intestinal (Caco-2) and Madin-Darby canine kidney (MDCK-1) polarized epithelial cells grown on a permeable support. Filter-grown Caco-2 cells and MDCK-1 cells, producing a transepithelial resistance of 300 to 500 and greater than 1,000 omega . cm2, respectively, were infected from either the apical or basolateral domain with RRV or Semliki Forest virus. Whereas Semliki Forest virus infection only occurred when input virions had access to the basolateral domain of MDCK-1 or Caco-2 cells, RRV infected MDCK-1 and Caco-2 monolayers in a symmetric manner. The effect of rotavirus infection on monolayer permeability was analyzed by measuring the transepithelial electrical resistance. Rotavirus infection on filter-grown Caco-2 cells caused a transmembrane leak at 18 h postinfection, before the development of the cytopathic effect (CPE) and extensive virus release. Electrical resistance was completely abolished between 24 and 36 h postinfection. Although no CPE could be detected on RRV-infected MDCK cells, the infection caused a transmembrane leak that totally abolished the electrical resistance at 18 to 24 h postinfection. Cell viability and the CPE analysis together with immunohistochemistry and immunofluorescence data indicated that the abolishment of resistance across the monolayer was due not to an effect on the plasma membrane of the cells but to an effect on the paracellular pathway limited by tight junctions. Attachment and penetration of rotavirus onto Caco-2 cells caused no measurable transmembrane leak during the first hour of infection. 相似文献
7.
We previously described the isolation and purification of two similar alpha 1-protease inhibitors from mouse plasma termed alpha 1-PI(E) and alpha 1-PI(T) because of their respective affinities for elastase and trypsin. Some of the biochemical and immunological properties of these proteins are reported. Both are acidic glycoproteins with pI's of 4.1-4.2. The plasma half-life of each inhibitor, determined after administration of the 125I-protein, is approximately 4 h both in normal mice and in mice after induction of the acute phase reaction. The two proteins have almost identical amino acid compositions and similar CNBr peptide maps. Tryptic maps, however, are considerably different. Reverse-phase chromatography separated alpha 1-PI(E) into three distinct isoforms, each eluting with approximately 60% acetonitrile. Under these conditions alpha 1-PI(T) shows a single peak, clearly different from those of alpha 1-PI(E). The three alpha 1-PI(E) isoforms have the same molecular weights on sodium dodecyl sulfate-gel electrophoresis and the same tripeptide sequence at their N-terminus, and appear to be immunologically identical. Polyclonal, monospecific antibodies to each native inhibitor, prepared in rabbits, showed no cross-reactivity when tested by functional assay or crossed immunoelectrophoresis. Interestingly, each antibody recognized epitopes on the C-terminal portion of its respective antigen. These studies confirm that alpha 1-PI(E) and alpha 1-PI(T), although highly similar, are products of different genes. Like human alpha 1-PI, the two mouse inhibitors are partially inactivated by mild oxidation with chloramine-T, losing all elastase inhibitor and lesser amounts of antichymotryptic and antitryptic activity. However, unlike the human protein, neither alpha 1-PI(E) nor alpha 1-PI(T) was found to have a methionine residue at its P1 site. 相似文献
8.
The nucleotide sequences of five finP alleles from various IncF plasmids (finP types I to V) as well as of three finP mutations were determined and compared. The finP gene specificity could be attributed to a variable, six-to-seven-nucleotide loop located between inverted repeats, and the sequence data were consistent with the product of finP being an RNA molecule rather than a protein. The finP mutations interrupted a proposed finP promoter or destabilized a predicted stem-and-loop structure in the finP RNA molecule. 相似文献
9.
Six embalmed human cadaveric hemi-pelves with their associated proximal femurs have been tested in vitro using 25 strain-gauge rosettes on each hemi-pelvis. Loads were applied up to 2.5 kN and principal stresses were computed from the principal strain data. Acetabular prostheses, cemented in place upon a cartilage-devoid but intact subchondral bone-plate, showed little change in stress-patterns when compared with the normal data, regardless of whether or not the component employed metal-backing. The use of 30 anchoring holes of 6.4 mm diameter, in the intact subchondral bone-plate, had little effect upon the stress-patterns, regardless of whether metal-backing was employed upon the prosthesis. When the subchondral bone-plate was removed, there were notable changes in the stress-pattern in the periacetabular region and on the medial wall of pelvis. The metal-backed prosthesis did not produce such notable changes as its plastic counterpart, when the subchondral bone-plate was removed. The use of a plastic prosthesis cemented in a Protrusio ring, in an acetabulum devoid of subchondral bone, produced notable changes in the stress-patterns in the whole periacetabular region and on the medial wall. 相似文献
10.
A total of 161 patients completed a questionnaire about their pattern of taking the oral contraceptive pill. Only 28% (45) of patients were taking the pill according to the manufacturer''s instructions, and in the event of the pill being missed only 26% of patients would use a sheath. A tenth of the patients believed that amenorrhoea always indicated pregnancy, but 35% believed that amenorrhoea was harmful to the body. This group did not differ in their pill taking from the remaining 65% of patients. 相似文献