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31.
32.
Summary Lamellar bodies are described in the non-ciliated epithelial bronchiolar cells of the normal mouse lung. They are constituted of smooth concentric membranes, with a cytoplasmic center. They are related to mitochondria. They seem to belong to smooth endoplasmic reticulum. An origin from Golgi elements is discussed.
Acknowledgment. This work was supported by Grant No 69088 of Conseil de la Recherche Médicale du Québec. 相似文献
33.
R. Joplin A. J. Strain J. M. Neuberger 《In vitro cellular & developmental biology. Plant》1989,25(12):1189-1192
Summary Biliary epithelial cells (BEC) lining the intra-hepatic biliary ducts are the site of damage in several immunologically mediated
liver diseases. BEC are difficult to isolate since they represent only 5% of the total cell number in normal liver. In this
communication, a novel method for their isolation from normal liver is presented using a monoclonal antibody (HEA125) with
specificity for an epithelial cell surface glyco-protein reported to be expressed in liver only by biliary epithelium. By
combining differential density centrifugation and immuno-magnetic separation using HEA125 pure BEC (105 cells/g fresh tissue) were prepared routinely. These cells were maintained in culture for up to 4 weeks with significant
increases in cell numbers. The ability to prepare BEC from human liver offers an opportunity to develop In Vitro models to
investigate the aetiology of diseases in intra-hepatic biliary epithelium.
EDITOR’S STATEMENT This is a novel application to purification of specific liver cell types directly from tissue. It is well-suited
for rapid communication because of its novelty and potential utility to investigators. 相似文献
34.
James C. Beck Howard L. Hosick Bruce A. Watkins 《In vitro cellular & developmental biology. Plant》1989,25(5):409-418
Summary We investigated the effects of conditioned media derived from mouse mammary fat pads on the proliferation of CL-S1 cells,
an epithelial cell line originally isolated from a preneoplastic mammary outgrowth line. Cell proliferation in vitro in serum-free
defined medium was compared to that in this medium conditioned using intact mammary fat pad pieces or isolated fat pad adipocytes.
Culture medium was conditioned by incubating the conditioning material in defined culture medium for 24 h at 37°C. Conditioned
medium induced CL-S1 proliferation as much as 10- to 20-fold above the minimal levels of growth in control cultures after
13 d of culture. The growth-stimulatory factor(s) had an apparent molecular weight of greater than 10 kDa. This growth-stimulatory
activity was both heat and trypsin stable. Because the role of adipose tissue is to store and release lipids, we next tested
whether lipids are released during medium conditioning. The lipid composition of the fat pad conditioned medium was characterized
using both thin layer and gas liquid chromatography. These lipid analyses indicated that the fat pad pieces released significant
amounts of fatty acids and phospholipids into the medium during the conditioning period. The free fatty acid composition included
both saturated and unsaturated molecules, and about 80% of the total fatty acids consisted of palmitate, stearate, oleate,
and linoleate. These same fatty acids were a structural component of the majority of phospholipid found in the medium. The
addition of palmitate or stearate to defined medium had no effect or was inhibitory for CL-S1 proliferation, depending on
the concentration used. Defined medium supplemented with oleate, arachidonate, or linoleate induced CL-S1 proliferation, and
the inhibitory effects of palmitate and stearate were overcome by addition of oleate and linoleate. These data indicate that
both unsaturated and saturated fatty acids are released from intact adipose cells of the mouse mammary fat pad and that fatty
acids can influence the growth of prenoplastic mouse mammary epithelium. Thus, unsaturated fatty acids, perhaps in conjunction
with other substances released simultaneously, are candidate molecules for the substances that mediate the effect of adipose
tissue on growth of epithelium.
This work was supported in part by a grant from the American Institute for Cancer Research; grant CA 46885 from the National
Institutes of Health, Bethesda, MD; and by State of Washington initiative 171. 相似文献
35.
Expression of the gap junction protein connexin43 in embryonic chick lens: Molecular cloning,ultrastructural localization,and post-translational phosphorylation 总被引:13,自引:0,他引:13
Linda S. Musil Eric C. Beyer Daniel A. Goodenough 《The Journal of membrane biology》1990,116(2):163-175
Summary Lens epithelial cells are physiologically coupled to each other and to the lens fibers by an extensive network of intercellular gap junctions. In the rat, the epithelial-epithelial junctions appear to contain connexin43, a member of the connexin family of gap junction proteins. Limitations on the use of rodent lenses for the study of gap junction formation and regulation led us to examine the expression of connexin43 in embryonic chick lenses. We report here that chick connexin43 is remarkably similar to its rat counterpart in primary amino acid sequence and in several key structural features as deduced by molecular cDNA cloning. The cross-reactivity of an anti-rat connexin43 serum with chick connexin43 permitted definitive immunocytochemical localization of chick connexin43 to lens epithelial gap junctional plaques and examination of the biosynthesis of connexin43 by metabolic radiolabeling and immunoprecipitation. We show that chick lens cells synthesize connexin43 as a single, 42-kD species that is efficiently posttranslationally converted to a 45-kD form. Metabolic labeling of connexin43 with32P-orthophosphate combined with dephosphorylation experiments reveals that this shift in apparent molecular weight is due solely to phosphorylation. These results indicate that embryonic chick lens is an appropriate system for the study of connexin43 biosynthesis and demonstrate for the first time that connexin43 is a phosphoprotein. 相似文献
36.
Summary Interactions between epithelial cells and their environment are critical for normal function. Mammary epithelial cells require hormonal and extracellular matrix (ECM) signalling for the expression of tissue specific characteristics. With regard to ECM, cultured mammary epithelial cells synthesize and secrete milk proteins on stromal collagen I matrices. The onset of function coincides both with morphogenesis of a polarized epithelium and with deposition of basement membrane ECM basal to the cell layer. Mammary specific morphogenesis and biochemical differentiation is induced if mammary cells are cultured directly on exogenous basement membrane (EHS). Thus ECM may effect function by the concerted effect of permissivity for cell shape changes and the direct biochemical signalling of basement membrane molecules.A model is discussed where initial ECM control of mammary epithelial cell function originates in the interstitial matrix of stroma and subsequently transfers to the basement membrane when the epithelial cells have accumulated and deposited an organized basement membrane matrix.Dedicated to Professor Stuart Patton on the occasion of his 70th birthday. 相似文献
37.
We have tested the ability of several B2 antagonists on the responses of the open-circuited isolated canine tracheal epithelium to the luminal addition of Bradykinin (BK), Lys-BK, and substance P (SP). All three peptides produced biphasic changes in transmural potential difference (PD), an initial decrease (dip) followed by an increase (rise). The B2 antagonists
-Argo [Hyp3,Thi5,8,
-Phe7]BK (B5630) reversibly inhibited both the dips and the rise with IC50 values of 2.01 · 10−8 and 1.54 · 10−7 M, respectively. The responses to SP were unaffected even with high concentrations of the antagonist. Other antagonists tested [
-Phe1,7,Thi5,8]BK (B4158), [
-Phe2,7]BK (B4404), and [
-Phe7,Hyp8]BK (B5092) were ineffective. 相似文献
38.
We used whole-cell patch-clamp recording techniques to investigate G protein-activated currents in cultured rat retinal pigment
epithelial (RPE) cells. Using 140 mm KCl intracellular and 130 mm NaCl extracellular solutions, rat RPE cells possessed both inward and outward K+ currents. Upon addition of the nonhydrolyzable guanine triphosphate analogue, guanosine-5′-O-(3-thiophosphate) (GTPγS, 0.1
mm), to the recording electrode, a nonspecific cation (NSC) current was elicited. The NSC current had a mean reversal potential
of +5.7 mV in 130 mm extracellular NaCl with Cs+-aspartate in the pipette, and was not affected by alterations in the extracellular Ca2+ or Cl− concentration. The GTPγS-activated current was found to be permeable to several monovalent cations (K+, Na+, choline, TRIS, and NMDG). Addition of fluoroaluminate, an activator of large molecular weight heterotrimeric GTP-binding
proteins (G proteins), to the intracellular recording solution activated the NSC current. The G protein involved was pertussis
toxin (PTX)-sensitive, since GTPγS failed to activate the NSC current in cells pretreated with PTX. Further investigation
of second messenger molecules suggested that activation of the NSC current was not affected by alterations in intracellular
Ca2+ or ATP. From these results, we conclude that a G protein-regulated NSC current is present in rat RPE cells. Activation of
the NSC current may sufficiently depolarize RPE cells to activate outward K+ currents. This would provide a mechanism by which these cells could rid themselves of accumulated K+.
Received: 25 January 1996/Revised: 24 April 1996 相似文献
39.
Yuichi Mazaki Makoto Mochii Ryuji Kodama Goro Eguchi 《Development, growth & differentiation》1996,38(4):429-437
When retinal pigmented epithelial cells (PEC) of chick embryos are cultured under appropriate conditions, the phenotype changes to that of lens cells through a process known as transdifferentiation. The first half of the process, characterized by dedifferentiation of PEC, is accompanied by a marked decrease in adhesiveness of PEC to collagen type I- or type IV-coated dishes. To understand the underlying mechanisms of this change, we analyzed the expression of integrins, which are major receptors for extracellular matrix components. Northern blot analysis with cDNA probes for chicken α3, α6, α8, αv, β1 and β5 integrin mRNA showed that the genes for all these integrins are transcribed at similar levels in PEC and dedifferentiated PEC (dePEC). Further analysis of β1 integrin, which is a major component of integrin heterodimers, showed that although the protein amount of β1 integrin was not changed, its localization at focal contacts seen in PEC was lost in dePEC. When anti-β1 integrin antibody was added to the PEC culture medium, a decrease of cell-substrate adhesiveness occurred, followed by a gradual change in both morphology and gene expression patterns to ones similar to those of dePEC. These findings suggest that an appropriate distribution of β1 integrin plays an essential role in maintaining the differentiated state of PEC through cell-substrate adhesion. 相似文献
40.
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