全文获取类型
收费全文 | 7814篇 |
免费 | 491篇 |
国内免费 | 297篇 |
出版年
2023年 | 171篇 |
2022年 | 240篇 |
2021年 | 288篇 |
2020年 | 205篇 |
2019年 | 252篇 |
2018年 | 292篇 |
2017年 | 174篇 |
2016年 | 197篇 |
2015年 | 241篇 |
2014年 | 486篇 |
2013年 | 585篇 |
2012年 | 436篇 |
2011年 | 419篇 |
2010年 | 359篇 |
2009年 | 370篇 |
2008年 | 412篇 |
2007年 | 421篇 |
2006年 | 339篇 |
2005年 | 289篇 |
2004年 | 270篇 |
2003年 | 229篇 |
2002年 | 194篇 |
2001年 | 134篇 |
2000年 | 116篇 |
1999年 | 121篇 |
1998年 | 112篇 |
1997年 | 122篇 |
1996年 | 93篇 |
1995年 | 93篇 |
1994年 | 93篇 |
1993年 | 84篇 |
1992年 | 70篇 |
1991年 | 69篇 |
1990年 | 44篇 |
1989年 | 40篇 |
1988年 | 41篇 |
1987年 | 24篇 |
1986年 | 28篇 |
1985年 | 40篇 |
1984年 | 49篇 |
1983年 | 31篇 |
1982年 | 46篇 |
1981年 | 49篇 |
1980年 | 45篇 |
1979年 | 35篇 |
1978年 | 29篇 |
1977年 | 19篇 |
1976年 | 24篇 |
1975年 | 23篇 |
1974年 | 23篇 |
排序方式: 共有8602条查询结果,搜索用时 15 毫秒
71.
Mitogen- and isoproterenol-induced changes of [Ca2+]i in T cells attached to a glass substrate were examined. Murine (C57BL/6) splenic T cells were attached to coverslips or 35-mm dishes (MatTek) precoated with Cell Tak® (3.5 µg/cm2). The cells were then loaded with fluorescent dye (2 µg/ml of fura2-AM or fluo3-AM) and changes in [Ca2+]i in a population of cells (using a spectrofluorometer) or in single cells (using a confocal microscope) were measured during continuous superfusion. Population measurements of [Ca2+]i demonstrated that concanavalin A (Con A, 2 or 5 µg/ml) caused an increase in [Ca2+]i that rose to a peak and then declined to a steady state. The concentration-response relationship (0.05–5 µg/ml) had an EC50 of 0.3 µg/ml. Isoproterenol decreased the Con A-induced elevation of steady state [Ca2+]i. In single cell studies, the increase in [Ca2+]i in response to Con A typically occurred in about 50% of the cells in a microscope field, and the delay before activation varied among cells. Taken together, these data demonstrate that Cell Tak® can be used to attach T cells to glass coverslips and will be useful for the study of signaling mechanisms in T cells. 相似文献
72.
Identification of Immunogenic Epitopes of the 170-kDa Subunit Adhesin of Entamoeba histolytica in Patients with Invasive Amebiasis 总被引:2,自引:0,他引:2
CARLOS VELAZQUEZ IGNEZ VALETTE MIGUEL CRUZ MARIA-LUISA LABRA JULIO MONTES SAMUEL L. STANLEY Jr JESUS CALDERON 《The Journal of eukaryotic microbiology》1995,42(5):636-641
ABSTRACT. Entamoeba histolytica causes amebic dysentery (AD) and liver abscess (ALA). Little is known about protective immunity to amebiasis, and studies in this area have been complicated by the paucity of defined ameba antigens. We examined the proliferative responses of peripheral blood mononuclear cells (PBMC) from patients with AD and ALA to a recombinant protein containing a portion of the 170 kDa adhesin of E. histolytica (170CR), and to two synthetic peptides (1 and 2) derived from the 170 kDa sequence that were predicted to contain T cell epitopes. A significant number of patients with AD and ALA had PBMC that proliferated to 170CR molecule, and several individuals with ALA and AD had T cells that recognized one or both peptides. Contrarily, individuals from a non-endemic region for amebiasis did not respond to 170CR protein, or to both peptides. In regard to antibody response, nine of fifteen patients with ALA showed antibodies to 170CR protein. These same patients had antibodies to peptide 2. We identified peptides from 170-kDa adhesin that may contain both T and B cell epitopes recognized by some patients with invasive amebiasis. These peptides may be valuable reagents in studies of the immune response to amebiasis. 相似文献
73.
Ali Arslan Cuillermina Almazan Hans H. Zingg 《In vitro cellular & developmental biology. Animal》1995,31(2):140-148
Summary Normal and neoplastic growth of epithelial cells depends on mutual interactions between epithelial and stromal cells. As a
tool for the study of the underlying molecular mechanisms, we have developed temperature-sensitive, nontransformed cell lines
derived from rat uterine epithelium and stroma by transfecting primary cultures with a temperature-sensitive mutant of the
SV40 large T antigen. The epithelial and stromal cell lines obtained shared relevant morphological characteristics with the
primary cells from which they were derived. Immunocytochemical analysis showed that the epithelial cell lines expressed the
intermediate filament cytokeratin, whereas the stromal lines expressed the intermediate filament vimentin. Alkaline phosphatase
activity was present in all cell lines examined. All cell lines were anchorage dependent and did not form foci. One epithelial
cell line expressed oxytocin mRNA, a gene product recently shown to be highly expressed in vivo in the uterine epithelium
at term. If grown on Matrigel, this cell line formed domelike structures, a further characteristic of its differentiated phenotype.
In an attempt to reconstitute an endometrium in vitro, epithelial cells were seeded on top of a layer of stromal cells. Paraffin
cross sections showed that this in vitro system consisted of a bilayer structure. Four to five cuboidal epithelial cells were
typically anchored atop one stromal cell, forming an endometriumlike tissue. The present in vitro system should provide a
useful model for further studies on endometrial functions and epithelial/stromal cell interactions at a molecular level. 相似文献
74.
Ana M. Celentano Gabriela Gorelik María E. Solana Leonor Sterin-Borda Enri Borda Stella M. Gonzlez Cappa 《Prostaglandins & other lipid mediators》1995,49(3)
PGE2 involvement in experimental Trypanosoma cruzi infection depends on the lethal capacity of the parasite subpopulation used. Mice acutely infected with non-lethal K98 displayed an enhancement in PGE2 serum levels during the acute period, while those infected with lethal T. cruzi subpopulations (RA or K98-2) showed levels not different from normal mice. The enhancement detected in K98 group could be related both to an increased number of CD8+ T cell number and to enhanced PGE2 release per cell by CD8+; values of PGE2 release by adherent cells were not altered in this group. Treatment with cyclooxygenase inhibitors enhanced mortality rates of mice infected with K98, and administration of 16,16-dimethyl PGE2 (dPGE) reversed this effect. However, mice infected with RA did not reduce their mortality rates by administration of diverse doses of dPGE. These findings suggest that PGE2 could play a role in resistance in mice infected with K98. 相似文献
75.
Eric Hbert Michel Monsigny 《Biology of the cell / under the auspices of the European Cell Biology Organization》1994,81(1):73-76
Summary— The increase in galectin-3 lectin content observed in tumours or in in vitro transformed cells suggests that this lectin is important in the transformation process. In the present study, we investigated the mRNA expression level of the galectin-3, galectin-I and macrophage mannose receptor in normal and ras-transformed NIH 3T3 cells in relation to their transformation state. The galectin3 mRNA content in ras-transformed cells is increased in fully transformed cells, with a maximum in ras-transformed cells that have lost their growth anchorage-dependence. Under the same conditions, the galectin-1 mRNA level which was high in normal cells, increased slightly in transformed cells. The mRNA for the macrophage mannose receptor was not detected in 3T3 cells or in their ras-transformed counterparts. 相似文献
76.
Munechika Enjoji 《Molecular and cellular biochemistry》1994,137(1):33-37
In the murine IgH gene intronic enhancer (ENHiH), two major functional domains were reported. One is the E4/octomer region and another includes the A and B motifs. In the human ENHiH, it was reported that the HE2, which corresponds to the murine B, and E6 motifs play an important role in an enhancer activity and a tissue-specificity at cellular level. Here we examined thein vivo function of the E6, A and HE2 motifs within the human ENHiH by using the transgenic mice technique. The A and HE2 motifs together revealed almost the same enhancer function as the whole human ENHiH, but the E6 motif had lesser enhancer acitivty and tissue-specificity. 相似文献
77.
P. D. Chen H. Tsujimoto B. S. Gill 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1994,88(1):97-101
Diploid-like chromosome pairing in polyploid wheat is controlled by several Ph (pairing homoeologous) genes with major and minor effects. Homoeologous pairing occurs in either the absence of these genes or their inhibition by genes from other species (Ph
I genes). We transferred Ph
I genes from Triticum speltoides (syn Aegilops speltoides) to T. aestivum, and on the basis of further analysis it appears that two duplicate and independent Ph
I genes were transferred. Since Ph
I genes are epistatic to the Ph genes of wheat, homoeologous pairing between the wheat and alien chromosomes occurs in the F1 hybrids. Using the Ph
I gene stock, we could demonstrate homoeologous pairing between the wheat and Haynaldia villosa chromosomes. Since homoeologous pairing occurs in F1 hybrids and no cytogenetic manipulation is needed, the Ph
I gene stock may be a versatile tool for effecting rapid and efficient alien genetic transfers to wheat.Contribution no. 93-435-J from the Kansas Agricultural Experiment Station, Kansas State University, Manhattan, KS 66506-5502, USA 相似文献
78.
Abstract: Derivatives of the lac promoter (tac, pac, rac) belong to the strongest bacterial promoters which are frequently used for the induced overexpression of foreign genes in Escherichia coli . However, their use in fermentation processes is strongly restricted because of the high cost of the inducer iso-propyl-β-D-thiogalactopyranoside (IPTG). The aim of this work was to investigate the possibility of using lac-derived promoters in high cell density processes resulting in a high yield of the induced recombinant protein if glucose is the main carbon and energy source. Lactose is tested as inducer of the main antigenic coat protein (VP1) of the foot and mouth disease (FMD) virus in a T7-RNA polymerase expression system. It was shown that lactose is able to induce the expression of the recombinant gene to an amount of the VP1 protein corresponding to 20% of the total cell protein. 相似文献
79.
Self-splicing group I and group II introns encode homologous (putative) DNA endonucleases of a new family. 总被引:17,自引:3,他引:14 下载免费PDF全文
A. E. Gorbalenya 《Protein science : a publication of the Protein Society》1994,3(7):1117-1120
A new family of protein domains consisting of 50-80 amino acid residues is described. It is composed of nearly 40 members, including domains encoded by plastid and phage group I introns; mitochondrial, plastid, and bacterial group II introns; eubacterial genomes and plasmids; and phages. The name "EX1HH-HX3H" was coined for both domain and family. It is based on 2 most prominent amino acid sequence motifs, each encompassing a pair of highly conserved histidine residues in a specific arrangement: EX1HH and HX3H. The "His" motifs often alternate with amino- and carboxy-terminal motifs of a new type of Zn-finger-like structure CX2,4CX29-54[CH]X2,3[CH]. The EX1HH-HX3H domain in eubacterial E2-type bacteriocins and in phage RB3 (wild variant of phage T4) product of the nrdB group I intron was reported to be essential for DNA endonuclease activity of these proteins. In other proteins, the EX1HH-HX3H domain is hypothesized to possess DNase activity as well. Presumably, this activity promotes movement (rearrangement) of group I and group II introns encoding the EX1HH-HX3H domain and other gene targets. In the case of Escherichia coli restrictase McrA and possibly several related proteins, it appears to mediate the restriction of alien DNA molecules. 相似文献
80.
旋毛虫肌幼虫ES抗原的基因克隆及高效表达 总被引:7,自引:0,他引:7
作者对编码旋毛虫肌幼虫ES抗原的部分结构基因进行了克隆、鉴定和表达。用RNA PCR技术直接从旋毛虫肌幼虫总RNA中反转录并扩增出0.7kh的靶DNA,酶切分析后将其克隆到融合表达载体pEx3lC中。SDS—PAGE电泳表明,含重组子的大肠杆菌能够表达出一分子量为37kDa的融合蛋白(P37),后者占菌体总蛋白的22%以上,并以包含体形式存在于菌体中。经对纯化后表达蛋白的ELlSA检测,证明它能被猪旋毛虫病阳性血清和抗旋毛虫单克隆抗体识别。研究结果揭示,重组蛋白P37对于研制旋毛虫病诊断抗原和免疫抗原具有潜在的应用价值。 相似文献