全文获取类型
收费全文 | 645篇 |
免费 | 46篇 |
专业分类
691篇 |
出版年
2021年 | 7篇 |
2020年 | 5篇 |
2019年 | 7篇 |
2018年 | 9篇 |
2017年 | 11篇 |
2016年 | 13篇 |
2015年 | 16篇 |
2014年 | 21篇 |
2013年 | 25篇 |
2012年 | 30篇 |
2011年 | 18篇 |
2010年 | 13篇 |
2009年 | 8篇 |
2008年 | 32篇 |
2007年 | 17篇 |
2006年 | 26篇 |
2005年 | 16篇 |
2004年 | 28篇 |
2003年 | 20篇 |
2002年 | 20篇 |
2001年 | 19篇 |
2000年 | 10篇 |
1999年 | 15篇 |
1997年 | 5篇 |
1996年 | 7篇 |
1995年 | 6篇 |
1994年 | 6篇 |
1993年 | 7篇 |
1992年 | 16篇 |
1991年 | 20篇 |
1990年 | 10篇 |
1989年 | 19篇 |
1988年 | 15篇 |
1987年 | 13篇 |
1986年 | 16篇 |
1985年 | 9篇 |
1984年 | 18篇 |
1983年 | 17篇 |
1982年 | 7篇 |
1981年 | 6篇 |
1980年 | 10篇 |
1979年 | 12篇 |
1977年 | 13篇 |
1976年 | 9篇 |
1975年 | 8篇 |
1974年 | 8篇 |
1973年 | 10篇 |
1969年 | 5篇 |
1967年 | 4篇 |
1966年 | 6篇 |
排序方式: 共有691条查询结果,搜索用时 0 毫秒
1.
Electron-microscopic and immunocytochemical analyses of Weibel-Palade bodies in the human umbilical vein during pregnancy 总被引:3,自引:0,他引:3
Summary The present study was done to elucidate the biological significance of the Weibel-Palade body of human umbilical vein endothelial cells. Quantitative determinations of these endothelial-specific granules throughout pregnancy revealed that their numbers and size per cell profile were maintained at low levels from 12 to 19 weeks of gestation; then both rapidly increased from 33 weeks to full term. This increase coincided with the development of the rough endoplasmic reticulum and an increase in the number of endothelial cell pinocytotic vesicles. Light-microscopic peroxidase anti-peroxidase and electron-microscopic protein A-gold techniques provided evidence that factor VIII-related antigen was localized in the Weibel-Palade bodies. Furthermore, in vitro treatment of incubated umbilical vein tissue with compound 48/80, a histamine releaser, induced degranulation of Weibel-Palade bodies from the endothelium. The present study indicates that Weibel-Palade bodies are storage sites of both histamine and factor VIII-related antigen and have an important role in the obliteration of this vessel. 相似文献
2.
Eleven temperature-sensitive mutants of adenovirus type 12, capable of forming plaques in human cells at 33 C but not at 39.5 C, were isolated from a stock of a wild-type strain after treatment with either nitrous acid or hydroxyl-amine. Complementation tests in doubly infected human cells permitted a tentative assignment of eight of these mutants to six complementation groups. Temperature-shift experiments revealed that one mutant is affected early and most of the other mutants are affected late. Only the early mutant, H12ts505, was temperature sensitive in viral DNA replication. Infectious virions of all the mutants except H12ts505 and two of the late mutants produced at 33 C, appeared to be more heat labile than those of the wild type. Only H12ts505 was temperature sensitive for the establishment of transformation of rat 3Y1 cells. One of the late mutants (H12ts504) had an increased transforming ability at the permissive temperature. Results of temperature-shift transformation experiments suggest that a viral function affected in H12ts505 is required for “initiation” of transformation. Some of the growth properties of H12ts505-transformed cells were also temperature dependent, suggesting that a functional expression of a gene mutated in H12ts505 is required to maintain at least some aspects of the transformed state. 相似文献
3.
M Nishioka T Aibiki M Shirai S Terada H Kagawa S Watanabe 《Microbiology and immunology》1986,30(12):1291-1297
Actin is a major antigen involved in the reaction of smooth muscle antibody positive sera from patients with chronic active hepatitis. In the present study, actin extracted from rabbit skeletal muscle was denatured by sodium dodecyl sulfate and was immunized into the rabbit, a homologous animal for actin. The rabbits, thus immunized, produced antibodies reactive with actins of homologous and heterologous animals. In addition, the antibodies showed reactivity with autologous actin. It indicates that the denatured homologous actin is capable of terminating immunological tolerance to actin and induces formation of autoantibody to rabbit actin. This phenomenon may be implicated in the occurrence of anti-actin antibody in sera from patients with chronic liver disease and several other diseases. 相似文献
4.
M Ohtsubo M Yoshida S Ohta Y Kagawa M Yohda T Date 《Biochemical and biophysical research communications》1987,146(2):705-710
Using site-directed mutagenesis, Glu-190 or Glu-201 of the beta subunit of the F1-ATPase from the thermophilic bacterium PS3 were replaced with glutamine. It was possible to reconstitute complexes of the mutated beta subunits with alpha and gamma subunits, but the complexes did not have ATPase activity. It is concluded that carboxylic acid side chains of Glu-190 and Glu-201 of the beta subunit are essential for catalytic activity of F1-ATPase. 相似文献
5.
A potential approach for gene therapy targeting hepatoma using a liver-specific promoter on a retroviral vector. 总被引:7,自引:0,他引:7
S Kuriyama M Yoshikawa S Ishizaka T Tsujii K Ikenaka T Kagawa N Morita K Mikoshiba 《Cell structure and function》1991,16(6):503-510
Recent technological advances made in molecular biology and in vitro culture of human and other mammalian cells have led to broad medical and scientific acceptance of the feasibility of gene therapy for genetic diseases. Cancer might practically be one of the attractive targets for such therapy. For the treatment of cancer, it is important to manipulate the gene of interest such that it is expressed solely in cancer cells. We have developed a tissue-specific gene expression system, based on a tissue-specific promoter on a retroviral vector. A murine ecotropic retroviral vector was constructed in which the Escherichia coli beta-galactosidase gene served as a reporter; it was expressed under control of the albumin enhancer element and promoter. The tissue specificity of this vector was first assessed in vitro, and beta-galactosidase activity was detected exclusively in hepatoma cell lines. This recombinant retrovirus was injected directly into a subcutaneous tumor composed of transplantable murine MH-134 hepatoma cells, and expression of the gene was observed in vivo. Then this recombinant retrovirus was injected via the spleen or directly into the liver, resulting in the gene expression in dividing hepatocytes in partially hepatectomized mice, but not in nondividing hepatocytes in normal mice. Gene transfer specific to dividing hepatocytes and expression by means of retroviral vectors should possess high potential for selective elimination of hepatoma cells surrounded by nondividing normal hepatocytes. 相似文献
6.
Sequence and over-expression of subunits of adenosine triphosphate synthase in thermophilic bacterium PS3 总被引:8,自引:0,他引:8
S Ohta M Yohda M Ishizuka H Hirata T Hamamoto Y Otawara-Hamamoto K Matsuda Y Kagawa 《Biochimica et biophysica acta》1988,933(1):141-155
The primary structures of all the subunits of thermophilic ATP synthase were determined, and its alpha, beta and gamma subunits could be over-expressed in Escherichia coli, because these subunits were stable and reconstitutable. DNA of 7500 base pairs in length was found to contain a cluster of nine genes for subunits of ATP synthase. The order of their reading frames (size in base pairs) was: I(381): a(630): c(216): b(489): delta(537): alpha(1507): gamma(858): beta(1419): epsilon(396), I being a gene for a small hydrophobic, basic protein expressed in vitro. All the termini of TF0F1 subunits were confirmed by peptide sequencing. Large quantities of the overexpressed thermophilic alpha, beta and gamma subunits were prepared from the extract of E. coli, by a few purification steps. 相似文献
7.
Highly purified sarcolemmal membranes, prepared from fresh bovine heart left ventricle, were solubilized by n-octyl beta-D-glucopyranoside and reconstituted into proteoliposomes with soybean phospholipids by the detergent-dialysis method. Ca2+ flux into the proteoliposomes was determined using the fluorescent probe Quin2. A membrane potential (negative in the proteoliposome interior) that was created by K+ diffusion mediated by valinomycin accelerated the Ca2+ influx. The voltage-dependent Ca2+ influx was dependent on pretreatment of the sarcolemmal membranes with Bay K 8644 and was inhibited by various calcium antagonists including nicardipine (K0.5 = 4.5.10(-7) M), verapamil (K0.5 = 9.2.10(-9) M), diltiazem (K0.5 = 26.10(-8) M) and omega-conotoxin (K0.5 = 9.5.10(-9) M). 相似文献
8.
F1-ATPase is the major enzyme for ATP synthesis in mitochondria, chloroplasts, and bacterial plasma membranes. F1-ATPase obtained from thermophilic bacterium PS3 (TF1) is the only ATPase which can be reconstituted from its primary structure. Its beta subunit constitutes the catalytic site, and is capable of forming hybrid F1's with E. coli alpha and gamma subunits. Since the stability of TF1 resides in its primary structure, we cloned a gene coding for TF1, and the primary structure of the beta subunit was deduced from the nucleotide sequence of the gene to compare the sequence with those of beta's of three major categories of F1's; prokaryotic membranes, chloroplasts, and mitochondria. The following results were obtained. Homology: The primary structure of the TF1 beta subunit (473 residues, Mr = 51,995.6) showed 89.3% homology with 270 residues which are identical in the beta subunits from human mitochondria, spinach chloroplasts, and E. coli. It contained regions homologous to several nucleotide-binding proteins. Secondary structure: The deduced alpha-helical (30.1%) and beta-sheet (22.3%) contents were consistent with those determined from the circular dichroism spectra. Residues forming reverse turns (Gly and Pro) were highly conserved among the F1 beta subunits. Substituted residues and stability of TF1: We compared the amino acid sequence of the TF1 beta subunit with those of the other F1 beta subunits mentioned above. The observed substitutions in the thermophilic subunit increased its propensities to form secondary structures, and its external polarity to form tertiary structure. Codon usage: The codon usage of the TF1 beta gene was found to be unique. The changes in codons that achieved these amino acid substitutions were much larger than those caused by minimal mutations, and the third letters of the optimal codons were either guanine or cytosine, except in codons for Gln, Lys, and Glu. 相似文献
9.
Koji Yamada Masafuni Sasaki Genki Kimura 《In vitro cellular & developmental biology. Plant》1986,22(4):212-216
Summary We examined cellular protein content in four temperature-sensitive (ts) mutants of rat 3Y1 fibroblasts (3Y1tsD123, 3Y1tsF121,
3Y1tsG125, and 3Y1tsH203) under various conditions of culture that affect cell proliferation. When proliferation of the ts
mutants was inhibited at a nonpermissive temperature (39.8°C) in the G1 phase, prominent accumulation of cellular protein occurred in three mutants (3Y1tsF121, 3Y1tsG125, and 3Y1tsH203) but not
in 3Y1tsD123. The over-accumulation of protein at 39.8°C in the former three mutants was inhibited at high cell densities.
At low cell densities there was an upper limit in the protein accumulation at 39.8°C. When the three mutants, proliferation-arrested
at high cell densities at 33.8°C, were replated sparsely in fresh medium and shifted to 39.8°C, proliferation was completely
inhibited whereas over-accumulation of protein occurred. These results indicating dissociation of protein accumulation and
cell proliferation suggest that the two events are regulated by different mechanisms.
This work was supported in part by a Grant-in-Aid for Encouragement of Young Scientists (1984) to K. Y. from the Ministry
of Education, Science, and Culture, Japan. 相似文献
10.
Resistance of thermophilic ATPase (TF1) to specific F1-atpase inhibitors including local anesthetics 总被引:2,自引:0,他引:2
T Saishu Y Kagawa R Shimizu 《Biochemical and biophysical research communications》1983,112(3):822-826
F1-ATPase obtained from mesophilic organisms is inhibited by specific inhibitors, such as aurovertin, efrapeptin, quercetin and several local anesthetics. This property has been explained by the common structure at the catalytic center of F1. However thermophilic F1 (TF1), which has the same primary structure at the center as other F1's, was shown to be resistant to these F1-specific inhibitors. Thus, the inhibitory mechanism may be explained not by the common structure at the catalytic site, but by some conformational changes of the flexible mesophilic F1 molecules or the absence of an inhibitor binding site in thermophilic F1. 相似文献