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排序方式: 共有86条查询结果,搜索用时 484 毫秒
21.
Characterization of entry mechanisms of human herpesvirus 8 by using an Rta-dependent reporter cell line 总被引:6,自引:0,他引:6
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To analyze the mechanisms of entry of human herpesvirus 8 (HHV-8), we established a reporter cell line T1H6 that contains the lacZ gene under the control of the polyadenylated nuclear RNA promoter, known to be strongly activated by a viral transactivator, Rta. We found that infection with cell-free virus, as well as cocultivation with HHV-8-positive primary effusion lymphoma cell lines, activated the lacZ gene of T1H6 in a sensitive and dose-dependent manner. Addition of Polybrene and centrifugation enhanced, but polysulfonate compounds inhibited, the HHV-8 infectivity. RGD-motif-containing polypeptides and integrins did not decrease the infectivity, suggesting the presence of an additional cellular receptor other than the reported one. The entry was dependent on pH acidification but not on the clathrin pathway. Although conditioned media obtained from human immunodeficiency virus (HIV)-infected cells did not have any effect on the early steps of HHV-8 infection, intracellular expression of a proviral HIV type 1, but not of Tat alone, increased the HHV-8-dependent reporter activation slightly, suggesting a potential of HIV-mediated enhancement of an early step of HHV-8 infection. 相似文献
22.
23.
Fabian Emrich Kiril Penov Mamoru Arakawa Nathan Dhablania Grayson Burdon Albert J. Pedroza Tiffany K. Koyano Young M. Kim Uwe Raaz Andrew J. Connolly Cristiana Iosef Michael P. Fischbein 《Journal of cellular and molecular medicine》2019,23(10):7000-7009
Marfan syndrome (MFS) is a connective tissue disorder that results in aortic root aneurysm formation. Reactive oxygen species (ROS) seem to play a role in aortic wall remodelling in MFS, although the mechanism remains unknown. MFS Fbn1C1039G/+ mouse root/ascending (AS) and descending (DES) aortic samples were examined using DHE staining, lucigenin‐enhanced chemiluminescence (LGCL), Verhoeff's elastin‐Van Gieson staining (elastin breakdown) and in situ zymography for protease activity. Fbn1C1039G/+ AS‐ or DES‐derived smooth muscle cells (SMC) were treated with anti‐TGF‐β antibody, angiotensin II (AngII), anti‐TGF‐β antibody + AngII, or isotype control. ROS were detected during early aneurysm formation in the Fbn1C1039G/+ AS aorta, but absent in normal‐sized DES aorta. Fbn1C1039G/+ mice treated with the unspecific NADPH oxidase inhibitor, apocynin reduced AS aneurysm formation, with attenuated elastin fragmentation. In situ zymography revealed apocynin treatment decreased protease activity. In vitro SMC studies showed Fbn1C1039G/+‐derived AS SMC had increased NADPH activity compared to DES‐derived SMC. AS SMC NADPH activity increased with AngII treatment and appeared TGF‐β dependent. In conclusion, ROS play a role in MFS aneurysm development and correspond anatomically with aneurysmal aortic segments. ROS inhibition via apocynin treatment attenuates MFS aneurysm progression. AngII enhances ROS production in MFS AS SMCs and is likely TGF‐β dependent. 相似文献
24.
Koji Yamano Bruno B. Queliconi Fumika Koyano Yasushi Saeki Takatsugu Hirokawa Keiji Tanaka Noriyuki Matsuda 《The Journal of biological chemistry》2015,290(42):25199-25211
Damaged mitochondria are eliminated through autophagy machinery. A cytosolic E3 ubiquitin ligase Parkin, a gene product mutated in familial Parkinsonism, is essential for this pathway. Recent progress has revealed that phosphorylation of both Parkin and ubiquitin at Ser65 by PINK1 are crucial for activation and recruitment of Parkin to the damaged mitochondria. However, the mechanism by which phosphorylated ubiquitin associates with and activates phosphorylated Parkin E3 ligase activity remains largely unknown. Here, we analyze interactions between phosphorylated forms of both Parkin and ubiquitin at a spatial resolution of the amino acid residue by site-specific photo-crosslinking. We reveal that the in-between-RING (IBR) domain along with RING1 domain of Parkin preferentially binds to ubiquitin in a phosphorylation-dependent manner. Furthermore, another approach, the Fluoppi (fluorescent-based technology detecting protein-protein interaction) assay, also showed that pathogenic mutations in these domains blocked interactions with phosphomimetic ubiquitin in mammalian cells. Molecular modeling based on the site-specific photo-crosslinking interaction map combined with mass spectrometry strongly suggests that a novel binding mechanism between Parkin and ubiquitin leads to a Parkin conformational change with subsequent activation of Parkin E3 ligase activity. 相似文献
25.
Algal phycocyanins promote growth of human cells in culture 总被引:4,自引:0,他引:4
K. Shinohara Y. Okura T. Koyano H. Murakami H. Omura 《In vitro cellular & developmental biology. Plant》1988,24(10):1057-1060
Summary The growth-promoting substances in a non-dialyzable extract of Synechococcus elongatus var. on RPMI 8226 cells (a human myeloma
cell line) were separated by gel filtration and ion exchange chromatography. By gel filtration with Sepharose 4B, the dialyzate
was separated into two fractions. One fraction was green-colored (P-1) and the other was blue-colored (P-2). The P-2 fraction
had a higher growth-promoting activity than P-1. By ion exchange chromatography, the P-2 fraction was separated into two blue-colored
fractions of phycocyanin and allophycocyanin. Both biliproteins promoted the growth of RPMI 8226 cells; however, allophycocyanin
was more active than phycocyanin.
Editor's statement This reporrt that phycobiliprotein from algae is capable of stimulating animal cell growth is unique, and
raises the possiblity that related compounds such as biliverdin might also be similarly active. Some puzzling aspects, such
as the lack of increase in specific activity during purification, as well as the possibility that the activity might be due
to a contaminant, remain to be resolved. 相似文献
26.
The catalytic domains of murine Golgi alpha1,2-mannosidases IA and IB that
are involved in N-glycan processing were expressed as secreted proteins in
P.pastoris . Recombinant mannosidases IA and IB both required divalent
cations for activity, were inhibited by deoxymannojirimycin and
kifunensine, and exhibited similar catalytic constants using
Manalpha1,2Manalpha-O-CH3as substrate. Mannosidase IA was purified as a 50
kDa catalytically active soluble fragment and shown to be an inverting
glycosidase. Recombinant mannosidases IA and IB were used to cleave
Man9GlcNAc and the isomers produced were identified by high performance
liquid chromatography and proton-nuclear magnetic resonance spectroscopy.
Man9GlcNAc was rapidly cleaved by both enzymes to Man6GlcNAc, followed by a
much slower conversion to Man5GlcNAc. The same isomers of Man7GlcNAc and
Man6GlcNAc were produced by both enzymes but different isomers of
Man8GlcNAc were formed. When Man8GlcNAc (Man8B isomer) was used as
substrate, rapid conversion to Man5GlcNAc was observed, and the same
oligosaccharide isomer intermediates were formed by both enzymes. These
results combined with proton-nuclear magnetic resonance spectroscopy data
demonstrate that it is the terminal alpha1, 2-mannose residue missing in
the Man8B isomer that is cleaved from Man9GlcNAc at a much slower rate.
When rat liver endoplasmic reticulum membrane extracts were incubated with
Man9GlcNAc2, Man8GlcNAc2was the major product and Man8B was the major
isomer. In contrast, rat liver Golgi membranes rapidly cleaved
Man9GlcNAc2to Man6GlcNAc2and more slowly to Man5GlcNAc2. In this case all
three isomers of Man8GlcNAc2were formed as intermediates, but a distinctive
isomer, Man8A, was predominant. Antiserum to recombinant mannosidase IA
immunoprecipitated an enzyme from Golgi extracts with the same specificity
as recombinant mannosidase IA. These immunodepleted membranes were enriched
in a Man9GlcNAc2to Man8GlcNAc2- cleaving activity forming predominantly the
Man8B isomer. These results suggest that mannosidases IA and IB in Golgi
membranes prefer the Man8B isomer generated by a complementary mannosidase
that removes a single mannose from Man9GlcNAc2.
相似文献
27.
A O Asita M Matsui T Nohmi A Matsuoka M Hayashi M Ishidate T Sofuni M Koyano H Matsushita 《Mutation research》1991,264(1):7-14
Smoke condensates of woods used for food preservation and aromatization in Nigeria were tested for mutagenic activity using Salmonella typhimurium TA98 and TA100. The woods were: white mangrove (Avicennia nitida), red mangrove (Rhizophora racemosa), mahogany Khaya sp.), abura (Mitragyna ciliata), alstonia (Alstonia boonei) and black afara (Terminalia ivorensis). Cigarette tar was tested for comparison. The condensates induced dose-dependent increases in the number of His+ revertants mainly with S9 mix. With the exception of mahogany and cigarette smoke condensate, the smoke condensates induced more revertants/microgram condensate in TA100 than in TA98. The number of revertants/microgram condensate ranged between 0.04 and 0.9 for the wood smoke condensates and was 0.12 for the cigarette smoke in TA100. The range was between 0.1 and 0.30 for the wood smoke condensates and 0.18 revertants/microgram condensate for cigarette smoke condensate in TA98. Concentrations of 7 polycyclic aromatic hydrocarbons (PAHs) in the condensates were determined namely, pyrene, benzo[a]pyrene, benz[a]anthracene, benzo[k]fluoranthene, benzo[b]chrysene, benzo[g,h,i]perylene and dibenzo[a,e]pyrene. The condensates contained varying concentrations of the individual PAHs and those with higher concentrations generally showed greater mutagenic activities. However, the order of mutagenic potency in the bacterial strains differed from the order of PAH concentrations, which were lower than the concentrations at which they are reported to induce mutations. When 6 of the PAHs were mixed in the concentrations in which they were found in the individual condensates, the mixtures did not induce mutation so that the contribution of the PAHs to the mutagenic activities of the condensates could not be determined. 相似文献
28.
29.
A peptide that blocks the interaction of NF‐κB p65 subunit with Smad4 enhances BMP2‐induced osteogenesis
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30.
We have developed a method for monitoring regional venous oxygen saturation. The key feature of this system is the use of highly flexible polymer fiber optics, and this flexibility allowed the production of a new fiber-optic transmission catheter. The space between the "face-to-face" positioned fiber-optic tips forms a remote catheter-based transmission cell. Our method applies Twersky's theory, in which absorption and scattering can be treated independently. Fresh rabbit blood was pumped through a disk oxygenator in which gas exchange occurred and passed the catheter. Simultaneous results obtained by the catheter and a cuvette oximeter were excellent (r = 0.99, SD = 1.1%). Oxygen saturation measured by this catheter was independent of vessel wall artifacts, blood pH, and flow velocity. Another application of this method is measurement of blood flow by the dye- (indocyanine green) dilution technique. The results of flow measurements by the catheter appeared to be satisfactory (r = 0.99, SD = 1.7%). This study concludes that our method is effective for monitoring the balance between regional oxygen supply and demand. 相似文献