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11.
Total cellular calcium content (determined by atomic absorption spectrometry) of Rat-1 cells transformed by temperature-sensitive Rous sarcoma virus decreases with cell density, but is found not significantly different at permissive and at non-permissive temperature. Kinetic analysis of 45Ca efflux from preloaded cells exhibits three separable pools of exchangeable calcium. The ratio of pool size of the fast-exchanging Ca-compartment (bound to cell surface) to pool size of the intermediate Ca-compartment (cytoplasmic) was found to decrease from 2.5 to 1.3 upon shift from non-permissive to permissive temperature. The slowly exchanging Ca-pool (presumably mitochondrial) did not change significantly upon temperature shift. These and further data demonstrate a close correlation between distribution of cellular Ca among different cellular compartments and characteristics of cellular proliferation, both attributable to the function(s) of a single oncogene. 相似文献
12.
13.
Expression of epidermal growth factor receptor and associated glycoprotein on cultured human brain tumor cells 总被引:3,自引:0,他引:3
P A Steck G E Gallick S A Maxwell W S Kloetzer R B Arlinghaus R P Moser J U Gutterman W K Yung 《Journal of cellular biochemistry》1986,32(1):1-10
The expression of epidermal growth factor (EGF-R) in normal glial and glioma cells grown in culture was examined by using several independent assays. Immunoprecipitation with the monoclonal antibody R1 of extracts from metabolically labeled glial and glioma cells revealed a protein of Mr approximately 170,000, with a migration in sodium dodecyl sulfate-polyacrylamide gels identical to the EGR-R of A431 epidermal carcinoma cells. Furthermore, in the majority of glioma extracts, a protein of Mr approximately 190,000 was specifically immunoprecipitated by this antibody. Similar results were obtained by immunoblotting with a second antibody directed against a synthetic peptide in the sequence of the v-erb-B oncogene. In cell lines expressing both proteins, each was specifically phosphorylated on tyrosine in immune complex kinase assays. The majority of glioma cells bound between 40,000 to 80,000 125I-labeled epidermal growth factor molecules per cell. These results suggest that the expression of EGF-R is common in cultured human glioma cells. In addition, a structurally related protein, is expressed in some of these cells. 相似文献
14.
Jila H. Boal Scott F. Deamond Daniel E. Callahan Sarah A. Bruce Paul O. P. Ts'o L. L. Kan 《Cell biochemistry and biophysics》1989,14(3):245-256
The nuclear magnetic resonance (NMR) parameters, spin-lattice (T1), and spin-spin (T2) relaxation time, are usually longer for neoplastic cells than for normal cells of the same cell type. This has generally
been true at low NMR frequencies (≤100 MHz) when comparisons have been made between normal and neoplastic cells that have
both spent a short time in culture. We have previously demonstrated that although the T1 values of paired normal and neoplastic Syrian hamster (SH) fibroblastic cells in culture are not significantly different
when measured at 300 MHz, the 300 MHz T2 values for the neoplastic cells are smaller than those of the normal cells. (Xin et al. (1986),Cell Biophysics
8, 213.) Since treatment of normal diploid cells with polypeptide growth factors or tumor promoters frequently results in reversible
expression of neoplasia-associated phenotypes, T1 and T2 were obtained at 300 MHz for treated and untreated SH cells to see if these compounds could also produce smaller 300 MHz
T2 values. Secondary culture SH fetal fibroblast cells were treated with epidermal growth factor (EGF), fibroblast growth factor
(FGF), phorbol-12,13-didecanoate (PDD) and 4-α-phorbol-12,13-didecanoate (4αPDD). Treatment with either growth factor resulted
in smaller T2 values, but a statistically significant decrease was not observed for PDD or 4αPDD. The observed reductions in T2 values were correlated with the morphological and growth-stimulatory effects of these compounds on the cells. 相似文献
15.
Summary The anastomosing ER system of epidermal cells of onion bulb scales is composed of three modifications: lamellar and tubular elements, located in the cell periphery, and long tubular stands located deeper in the cytoplasm. Cytoplasmic acidification of epidermal cells by loading with weak organic acids like acetic or propionic acid causes the decay of the lamellar elements and the disappearance of long tubular strands. Organelle movement is also inhibited. The effects depend on the pH of the incubation medium and on the administered acid concentration, and are characterized by a distinct lag phase of about 7 min. The induced ER changes are transient with adaptation starting after about 50min. Buffer components alone have little influence on the cellular ER organization within a pH-range of 4.0–8.0. However, the pH of the medium strongly affects the time course of the effects as well as recovery after omitting the administered acid. Both modulation and recovery occur more rapidly at neutral or slightly alkaline pH. Actin filaments, which play a major role in ER organization and organelle movement, are not affected by cytosolic acidification.Dedicated to the memory of Professor Oswald Kiermayer 相似文献
16.
By indirect immunofluorescence, using rabbit anti-heparin-binding placental protein (HBPP) antiserum, we studied HBPP expression by physiologically and non-physiologically (microsurgically) activated hamster gametes. Whereas mature gametes (sperm, metaphase II oocytes) were negative, in vivo conceived preimplantation embryos, from pronuclear to two- and four-cell stages, were HBPP positive. No HBPP was demonstrated in the zona pellucida, but HBPP-dependent immunofluorescence was localized in the perivitelline space. Oocytes incubated with hyaluronidase demonstrated variable responses from negative to positive. (Diluent or sperm) microinjected oocytes were all activated and HBPP positive within 4 h after stimulation. Thus neither activation by microinjection nor HBPP expression required paternal gametes. These kinetics suggest that HBPP may be a cortical granule secretogogue which can be applied to monitor oocyte responses during in vitro manipulations. 相似文献
17.
Angie Rizzino Peter Kazakoff John Nebelsick 《In vitro cellular & developmental biology. Plant》1990,26(5):537-542
Summary Previous studies have shown that cell density can regulate the binding of several growth factors. To determine whether cell
density exerts a uniform effect on the expression of epidermal growth factor (EGF) receptors, seven cell lines were examined
in detail. For each cell line, EGF binding was found to decrease as cell density increases. Scatchard analysis of the binding
data reveals that decreases in EGF binding are due to reductions in the number of cell surface EGF receptors. The only apparent
exception is the effect of cell density on the binding of EGF to A-431 cells. For these cells, increases in cell density lead
to two effects: decreases in the number of high affinity EGF receptors and increases in the total number of EGF receptors.
In addition to the effects of cell density on EGF receptors, it was determined that increases in cell density can coordinately
down-regulate receptors for as many as four different growth factors. Overall, the findings described in this report for EGF
and those previously described for transforming growth factor type-β (TGF-β) and fibroblast growth factor (FGF) demonstrate
the existence of a common mechanism for down-regulating growth factor receptors.
This work was supported by grants from the Nebraska Department of Health (89-51), the National Cancer Institute (Laboratory
Research Center Support Grant, CA36727), and the American Cancer Society (Core Grant ACS SIG-16).
EDITOR'S STATEMENT The paper by Rizzino et al. demonstrates that receptor number decreases as a function of cell density.
This may represent a mechanism by which cell proliferation is reduced as cell density increases. 相似文献
18.
Mouse submandibular salivary epithelial cell growth and differentiation in long-term culture: Influence of the extracellular matrix 总被引:7,自引:0,他引:7
Elisa M. Durban 《In vitro cellular & developmental biology. Plant》1990,26(1):33-43
Summary The adult mouse submandibular salivary gland provides a good model system to study gene regulation during normal and abnormal
cell behavior because it synthesizes functionally distinct products ranging from growth factors and digestive enzymes to factors
of relevance to homeostatic mechanisms. The present study describes the long-term growth and differentiation of submandibular
salivary epithelial cells from adult male mice as a function of the culture substratum. Using a two-step partial dissociation
procedure, it was possible to enrich for ductal cells of the granular convoluted tubules, the site of epidermal growth factor
synthesis. Long-term cell growth over a period of 2 to 3 mo. with at least 3 serial passages was obtained only within three-dimensional
collagen gels. Cells grew as ductal-type structures, many of which generated lumens with time in culture. Electron microscopic
analysis in reference to the submandibular gland in vivo revealed enrichment for and maintenance of morphologic features of
granular convoluted tubule cells. Reactivity with a keratin-specific monoclonal antibody established the epithelial nature
of the cells that grew within collagen. Maintenance of cell differentiation, using immunoreactivity for epidermal growth factor
as criterion, was determined by both cytochemical and biochemical approaches and was found to be dependent on the collagen
matrix and hormones. Greater than 50% of the cells in primary collagen cultures contained epidermal growth factor only in
the presence of testosterone and triiodothyronine. In contrast, cells initially seeded on plastic or cycled to plastic from
collagen gels were virtually negative for epidermal growth factor. Biochemical analysis confirmed the presence of a protein
with an apparent molecular weight of 6000 which comigrated with purified mouse epidermal growth factor. Epidermal growth factor
was also present in detectable levels in Passage 1 cells. This culture system should permit assessment of whether modulation
of submandibular gland ductal cell growth can be exerted via a mechanism that in itself includes epidermal growth factor and
its receptor and signal transduction pathway.
This work was supported by Public Health Service grant DE07766 from the National Institute of Dental Research, National Institutes
of Health, Bethesda, MD. 相似文献
19.
Toshimitsu Okeda Yasushi Yokogawa Hiroaki Ueo Mary A. Bury Paul O. P. Ts'o Sarah A. Bruce 《In vitro cellular & developmental biology. Plant》1990,26(12):1157-1166
Summary Primary cultures of 9-d-gestation Syrian hamster embryo (E9) cells are distinct from primary cultures of later gestational
age in terms of their growth and differentiation. First, primary E9 cell cultures express multiple mesenchymal differentiation
lineages (e.g., adipocyte, myoblast) only rarely seen in cultures of 13-d-gestation fetal (F13) cells. Second, although most
primary E9 cultures have a limited in vitro proliferative life span and exhibit cellular senescence similar to primary cultures
of F13 cells, E9 cultures seem to have higher frequency of escape from senescence and conversion to continuous cell lines
compared to F13 cells. Moreover, this frequency can be further increased 4- to 5-fold by continuous exposure of the E9 cells
to tumor promoters or epidermal growth factor. Eleven continuous cell lines have been isolated from unreated, promoter-treated,
or epidermal growth factor-treated primary E9 cultures. Seven of these are neoplastic or preneoplastic. However, the remaining
four do not show any evidence of being in neoplastic progression and three of these continue to express the same differentiated
phenotype observed in ther parental primary cell cultures.
These studies were supported in part by grants from the National Institutes of Health (AG 01998), Bethesda, MD, and the U.S.
Department of Energy (DE-A-C02-76-EVO-3280), Washington, DC. 相似文献
20.