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141.
Twisted ribbons made of polystyrene were used as a packing material for the cultivation of anchorage dependent cells. Normal human fibroblast cells grown on this support in a laboratory scale reactor reached densities of about 5–7×105 cells/ml. The cells adhered strongly to the carrier and no cell detachment was observed upon transfer to serum free medium. The properties of this packing material and its potential use are discussed. 相似文献
142.
Päivi Heikkilä Arvi I. Kahri Christian Ehnholm Petri T. Kovanen 《In vitro cellular & developmental biology. Plant》1988,24(9):936-942
Summary To define the role of endogenously synthesized cholesterol in the differentiation of adrenocortical cells in primary culture,
fetal rat adrenal cells were cultured in the presence of exogenous cholesterol (serum-supplemented medium) or in the absence
of it (serum-free medium or lipoprotein-free medium). Ultrastructurally the cells had features of glomerulosa cells: mitochondria
were oval or rod shaped with lamellar inner membranes. The amount of smooth endoplasmic reticulum was small, and lipid droplets
were few. When the cells were cultured in serum-free medium some intracytoplasmic vacuoles were seen. The undifferentiated
zona glomerulosa-like cells secreted low amounts of corticosterone and 18-OH-deoxycorticosterone (18-OH-DOC) in all three
media (serum-supplemented medium, serum-free medium, and lipoprotein-free medium). Stimulation of the adrenocortical cells
with ACTH induced the ultrastructural features of differentiated zona fasciculata-like cells. Mitochondrial inner membranes
were well developed in lipoprotein-free medium, but not in serum-free medium. The amount of intracellular lipids was increased
in both media devoid of cholesterol. In the ACTH stimulated cultures the presence of exogenous cholesterol resulted in increased
secretions of corticosterone and 18-OH-DOC. In the absence of an exogenous source of cholesterol, the amounts of steroids
secreted were only half of that secreted in the presence of serum-supplemented medium. Endogenously synthesized cholesterol
is sufficient for the morphologic differentiation of fetal rat adrenocortical cells under ACTH stimulation. However, without
exogenously provided cholesterol, the steroid production accounts only for half of the maximal output achieved using serum-supplemented
medium.
This work was supported by Finnish Culture Foundation. 相似文献
143.
A computerized mechanical cell stimulator for tissue culture: Effects on skeletal muscle organogenesis 总被引:8,自引:0,他引:8
Herman H. Vandenburgh 《In vitro cellular & developmental biology. Plant》1988,24(7):609-619
Summary A tissue culture system has been developed which can mechanically stimulate cells growing on a highly elastic plastic substratum
in a 24-well cell growth chamber. The collagen-coated substratum to which the cells attach and grow in the Mechanical Cell
Stimulator (Model I) can be repetitively stretched and relaxed by stepper motor with linear accuracy of 30 μm. The activity
controlling unit is an Apple IIe computer interfaced with the cell growth chamber via optical data links and is capable of
simulating many of the mechanical activity patterns that cells are subjected to in vivo. Primary avian skeletal myoblasts
proliferate and fuse into multinucleated myotubes in this set-up in a manner similar to normal tissue culture dishes. Under
static culture conditions, the muscle cells differentiate into networks of myotubes which show little orientation. Growing
the proliferating muscle cells on a unidirectional stretching substratum causes the developing myotubes to orient parallel
to the direction of movement. In contrast, growing the cells on a substratum undergoing continuous stretch-relaxation cycling
orients the developing myotubes perpendicular to the direction of movement. Neither type of mechanical activity significantly
affects the rate of cell proliferation of the rate of myoblast fusion into myotubes. These results indicate that during in
vivo skeletal muscle organogenesis, when substantial mechanical stresses are placed on skeletal muscle cells by both continuous
bone elongation and by spontaneous contractions, only bone elongation plays a significant role in proper fiber orientation
for subsequent functional work.
Supported by grants NS16753, AR36266, and RR05818 from the National Institutes of Health, Bethesda, MD. 相似文献
144.
Cyprian Weaver Robert L. Sorenson Brian Kobienia 《In vitro cellular & developmental biology. Plant》1988,24(2):108-116
Summary The purpose of this study was to develop a nonenzymatic method of isolating adult islets using atrophied pancreata from copper-deficient
rats and to analyze their morphologic characteristics and behavior in culture. This unusual model of isolation was studied
because islets remain intact in the course of dietary copper deficiency while the acinar glandular component of the pancreas
undergoes selective atrophy and lipomatosis. Small fragments containing islets were readily microdissected from atrophied
glands and placed in culture. Within 24 h the fragments congealed into small irregular- to spherical-shaped masses within
which the darker profile of islets could be distinguished. Within a period of 3 to 5 d, islet tissue began to bud from the
lipocytic mass until by Day 7 spherical aggregates of intact islet tissue separated from the residual fragments. Subsequent
to further in vitro treatment, these islets could be maintained as free viable spherical masses if periodically agitated,
as attached stationary islets which developed monolayer growth if left undisturbed and as aggregated masses of islet tissue
forming megaislets if combined in small groups. Grouped islets treated with actinomycin D and cycloheximide did not exhibit
aggregation when incubated with these inhibitors. This suggests that megaislet formation was an active process requiring protein-RNA
synthesis rather than passive clumping or aggregation that can accompany metabolically altered or dying islets undergoing
cellular shedding and adhesion. Immunohistochemical localization demonstrated that insulin, glucagon, somatostatin, and pancreatic
polypeptide-immunoreactive cell types were present within the islets derived from this technique. The cellular topography
of these islets was not unlike that described by others for islets cultured from enzymatic isolation. This culture model may
serve as a resource for mature, viable islets isolated without mechanical or enzymatic disaggregation which can have attenuating
effects on islet function.
This work was supported by a research grant from the Diabetes Research and Education Foundation. 相似文献
145.
M. Pilar López M. Jose Gómez-Lechón Jose V. Castell 《In vitro cellular & developmental biology. Plant》1988,24(6):511-517
Summary This study examines the factors involved in the rapid glycolysis and glycogenolysis that occur during the first stages of
hepatocyte culture: a) Shortly after seeding glycolysis, estimated as lactate released to culture medium, increased 10 times
in comparison to that reported in vivo. By 8 to 9 h of culture, hepatocytes were nearly glycogen-depleted even in the presence
of insulin. b) 6-Phosphofructo-2-kinase remained 100% active during this period. The proportion of the initial active phosphorylase
(87%) decreased to 57% by 7 h of culture. c) Fructose 2,6-bisphosphate content was initially similar to that found in liver
of fed animals, decreased after seeding and increased thereafter up to four times the initial concentration. In spite of changes
in the concentration of this activator, the glycolytic rate remained high and constant. d) ADP and AMP increased sharply after
cell plating, reaching values 1.7 and 3.5 times higher. The rise in AMP levels may be involved in the activation of glycolysis
and glycogenolysis, because this metabolite is known to act as an allosteric activator of phosphofrucktokinase and glycogen
phosphorylase. This metabolic situation resembles that of cells under hypoxia.
Part of this work was presented at the 38th Annual Meeting of the Tissue Culture Association, Washington, DC, May 1987. 相似文献
146.
Kiyoshi Akeo Yasuhiko Tanaka Tatsuji Fujiwara 《In vitro cellular & developmental biology. Plant》1988,24(7):705-710
Summary The endocytotic process in cultured human RPE cells was observed after 1 min, 20 min, and 2 h incubation with cationized ferritin.
Within 1 min the ferritin particles were seen to attach to the cell membrane, especially between microvilli. Uncoated and
coated pits could be recognized on the cell membranes, and uncoated and coated endocytotic vesicles were found in the cytoplasm
after 20 min of incubation. These vesicles were surrounded by abundant microfilaments and had no visible membranes. Loss of
membrane may be an initial step in the process of developing into the irregular clumps of ferritin particles found inside
the plasma membrane. With time, more irregular clumps of ferritin, smaller than the particles introduced during incubation,
appeared just beneath the cell membrane. Lysosomes were adjacent to the clumps of ferritin particles associated with microtobules
and finally degraded these particles. The phagolysosomes containing many particles were surrounded by many microtubules. Small
ferritin particles surrounded but had not entered the rough endoplasmic reticulums, and no particles were seen either around
or in the Golgi apparatus.
Presented at the 7th International Congress of Eye Research, Nagoya, Japan, 27 September 1986. 相似文献
147.
A. Birkenfeld Y. Ezra N. Ron D. Navot S. Granovsky J. G. Schenker I. S. Levij I. Vlodavsky 《In vitro cellular & developmental biology. Plant》1988,24(12):1188-1192
Summary The culturing of human endometrium in conventional plastic dishes and media is only partially successful, mainly because a
growth of a heterogeneous population of cells is achieved. Naturally produced extracellular matrix closely resembles the subepithelial
basement membrane and seems to affect both growth and differentiation of cells. These qualities of the extracellular matrix
(ECM) were applied for obtaining endometrial epithelial cultures. Endometrial tissue specimens were plated after slicing on
ECM-coated dishes and kept for up to 8 d. The growth of a confluent homogeneous tissue composed of polygonal epithelial-like
cells was demonstrated. To further characterize these cells, cultures were examined by scanning electron microscopy and transmission
electron microscopy. Scanning electron microscopy revealed flattened polygonal cells covered with microvilli, among which
ciliated cells were observed. By transmission electron microscopy the cells were seen as a monolayer, with some cells overlapping,
closely adherent to the matrix. Microvilli, as well as intracellular vacuoles and glycogen granules were observed. Cell type
specific cytoskeletal markers were demonstrated by antibodies to intermediate filament proteins (keratin and epithelial membrane
antigen). Taken together, the morphologic and immunohistochemical studies indicate that a selective growth of the epithelial
component of endometrial tissue was obtained after plating unprocessed endometrial tissue fragments on ECM-coated culture
dishes.
This work was supported by PHS grant no. CA 30289 to J.V. 相似文献
148.
A method for the rapid establishment of normal adult mammalian colonic epithelial cell cultures 总被引:6,自引:0,他引:6
Alda Vidrich Rajeswari Ravindranath Kianbanoo Farsi Stephan Targan 《In vitro cellular & developmental biology. Plant》1988,24(3):188-194
Summary Normal colonic epithelial cell cultures of mammalian origin are required to facilitate the study of both normal cellular functions
as well as pathogenesis of certain (human) colonic diseases. To date, little information is available regarding the growth
requirements of colonic epithelial cells in culture of eitehr animal or human origin. Such data would enable the development
of a long-term culture system for these cells. In this study, we present methodology that results in the establishment of
homogeneous cultures of adult rabbit colonic epithelial reproducibly, quickly, and in quantity. The epithelial nature of the
cultures is unambiguously established by intermediate filament typing using antikeratin antibodies. Such culutres can now
be used for a variety of functional studies as well as to investigate the growth requirements of colonic epithelial in culture.
This work was supported by the Blinder Foundation for Crohn’s Disease Research, Harbor UCLA IBD Center (AM 36200) and grant
AM 27806 from the National Institutes of Health, Bethesda, MD. 相似文献
149.
Takaki Shimada 《Cell and tissue research》1992,267(2):251-260
Summary Immunohistochemical localization of keratin, an intermediate filament protein, was studied in bull, goat, and sheep anterior pituitary glands, i.e., in animals of the order Artiodactyla. Strong immunoreactivity was detected in the cells of the marginal layer of bull and goat, as well as in cysts or large follicles in the anterior lobe of all 3 species. In addition, a number of stellateshape cells were immunoreactive for keratin and were distributed throughout the anterior lobe. The localization of keratin-positive cells in light-microscopic preparations correlated precisely with the localization of folliculo-stellate cells in adjacent ultrathin sections. In ultrastructural studies, many slender and elliptical membranous components which were different from smooth endoplasmic reticulum were observed in the cytoplasm of the some keratin-positive cells. Some of the folliculostellate cells in the 3 species were also immunoreactive for the subunit of S-100 protein, which exists in some epithelial cells. On the other hand, immunolocalization of glial fibrillary acidic protein, a glial cell marker, could not be demonstrated in the anterior pituitary glands of the 3 species studied. These results suggest that keratin-positive folliculo-stellate cells express epithelial-like characteristics. 相似文献
150.
P. Vuillez F. René M. Plante C. Hindelang M. J. Klein J. M. Félix M. E. Stoeckel 《Cell and tissue research》1992,267(1):169-183
Summary Organotypic cultures, in defined medium, of pituitary primordia obtained from 15-day-old rat fetuses were performed in order to study the in vitro differentiation of melanotrophic cells. The morphological and ultrastructural features of the transplants resembled those of the gland developing in vivo. In situ hybridization on semi-thin sections, using a 35S-labelled oligonucleotide probe, revealed pro-opiomelanocortin-mRNA-containing cells on the first day of culture in the anterior lobe and after 2–3 days in the intermediate lobe. Immunoperoxidase labelling of adjacent sections showed that the same cells reacted with antibodies against -melanocyte-stimulating hormone (MSH), 3 and adrenocorticotropic hormone in both lobes. The pro-opiomelanocortin-mRNA-containing cells formed progressively conspicuous areas in the intermediate lobe, which was almost uniformly labelled after 6 days. In the anterior lobe, these cells remained scattered in small cell groups, and colloidal gold immunolabelling showed the progressive disappearance of MSH labelling from the secretory vesicles in cells exhibiting morphological features of adult corticotrophic cells. Both the MSH content of the explants and MSH release into the culture medium increased with time. Treatment with the dopamine agonist bromocriptine induced a strong dose-dependent decrease in MSH secretion, which was significant after 3 days in culture, indicating that dopamine D2 receptors are able to regulate hormonal release of melanotrophic cells at early stages. This system constitutes a suitable model for further studies of factors controlling cell differentiation and cellular interactions involved in histogenesis. 相似文献