全文获取类型
收费全文 | 102315篇 |
免费 | 7604篇 |
国内免费 | 3578篇 |
出版年
2023年 | 1542篇 |
2022年 | 2023篇 |
2021年 | 3385篇 |
2020年 | 3500篇 |
2019年 | 4677篇 |
2018年 | 4025篇 |
2017年 | 2879篇 |
2016年 | 2835篇 |
2015年 | 3610篇 |
2014年 | 6502篇 |
2013年 | 7806篇 |
2012年 | 4743篇 |
2011年 | 6139篇 |
2010年 | 4585篇 |
2009年 | 5234篇 |
2008年 | 5380篇 |
2007年 | 5354篇 |
2006年 | 4891篇 |
2005年 | 4249篇 |
2004年 | 3781篇 |
2003年 | 3023篇 |
2002年 | 2678篇 |
2001年 | 1808篇 |
2000年 | 1507篇 |
1999年 | 1379篇 |
1998年 | 1429篇 |
1997年 | 1198篇 |
1996年 | 1147篇 |
1995年 | 1092篇 |
1994年 | 1019篇 |
1993年 | 836篇 |
1992年 | 865篇 |
1991年 | 728篇 |
1990年 | 656篇 |
1989年 | 558篇 |
1988年 | 489篇 |
1987年 | 465篇 |
1986年 | 383篇 |
1985年 | 545篇 |
1984年 | 717篇 |
1983年 | 470篇 |
1982年 | 580篇 |
1981年 | 459篇 |
1980年 | 373篇 |
1979年 | 368篇 |
1978年 | 286篇 |
1977年 | 253篇 |
1976年 | 220篇 |
1974年 | 143篇 |
1973年 | 181篇 |
排序方式: 共有10000条查询结果,搜索用时 15 毫秒
881.
882.
Cora-Jean S. Edgell Jill E. Haizlip C. Robert Bagnell Joan P. Packenham Paul Harrison Barry Wilbourn Victoria J. Madden 《In vitro cellular & developmental biology. Plant》1990,26(12):1167-1172
Summary Weibel-Palade bodies are ultrastructurally defined organelles found only in vascular endothelial cells. Because endothelium
in corpo is very dispersed, isolation and further characterization of this organelle has been dependent on increasing the
number of cells in culture. However, primary isolates of endothelial cells have a limited replication potential and tend to
senesce in culture. In this report, EA.hy926, a continuously replicating cell line derived from human endothelium, is shown
to contain Weibel-Palade bodies. Electron micrographs demonstrate the ultrastructural characteristics of these tissue-specific
organelles and their cytoplasmic distribution in EA.hy926 cells. Von Willebrand factor, which has been shown to exist in Weibel
Palade bodies, is demonstrated by immunofluorescence in discrete rod-shaped organelles whose size, shape, and distribution
are consistent with that of Weibel-Palade bodies in primary endothelial cell cultures. Rapid release of von Willebrand factor
can be induced by calcium ionophore, and large multimeric forms of the protein are found in EA.hy926 cells. These two properties
are consistent with the function currently ascribed to Weibel Palade bodies: storage of multimerized von Willebrand factor.
Thus ultrastructural, immunologic, and functional data establish the existence of this as yet poorly understood tissue-specific
organelle in a continuous, vigorously replicating human cell line. 相似文献
883.
Serum-free culture of enriched mouse anterior and ventral prostatic epithelial cells in collagen gel
Timothy Turner Howard A. Bern Peter Young Gerald R. Cunha 《In vitro cellular & developmental biology. Plant》1990,26(7):722-730
Summary Sustained growth of mouse ventral and anterior prostatic epithelial cells embedded within collagen gel matrix was achieved
in a serum-free medium composed of Dulbecco's modified Eagle's medium and Ham's F12 medium, 1∶1 (vol/vol), supplemented with
bovine serum albumin fraction V, epidermal growth factor, transferrin, cholera toxin, prolactin, 5α-dihydrotestosterone, cortisol,
putrescine, fibroblast growth factor, and a trace element mixture. Three-dimensional growth of prostatic epithelial cells
occurred inside the collagen gel matrix. This serum-free medium allowed cell growth greater than sevenfold over 10 d in culture.
Tissue recombination and cell culture techniques were integrated to demonstrate that cultured cells retained prostatic characteristics.
Following 10 d of culture, epithelial colonies from mouse ventral and anterior prostatic epithelial cell cultures were isolated
and combined with rat fetal urogenital sinus mesenchyme and grown for 4 wk under the renal capsule of intact athymic male
mice. These tissue recombinants showed distinctive prostatic histologic characteristic (alveoli and ducts lined with cuboidal
or columnar epithelium surrounded by stroma). When histologic sections of recombinants were stained with the Hoechst 33258,
epithelial cells of mouse origin were distinguishable from stromal cells of rat origin.
Aided by grants CA-05388 and CA-09041 from the National Institutes of Health, Bethesda, MD, and by M. A. R. C. fellowship
GM08730 to T. T. 相似文献
884.
Susan Mcafee Moore Coral A. Lamartiniere 《Journal of biochemical and molecular toxicology》1990,5(4):237-243
We have used the human hepatoma cell line, Hep G2, to examine the ability of hormones and xenobiotics to modulate the hepatic induction of benzo(a)pyrene hydroxylase and epoxide hydrolase. Hep G2 cells were cultured in Eagle's Minimum Essential Medium supplemented with 10% fetal calf serum. 3-Methylcholanthrene, diethylstilbestrol, testosterone propionate, and combinations of 3-meth-ylcholanthrene, and each of the hormones were added directly to the culture media. We subsequently studied the metabolism of benzo(a)pyrene using cell lysates of the Hep G2 cells. Metabolites were quantitated by high-performance liquid chromatography (HPLC) using fluorodetection. Exposure to 3-methyl-cholanthrene alone resulted in an eightfold increase in total benzo(a)pyrene metabolites with a change of the predominant metabolite from the 3-hydroxy-benzo(a)pyrene to the carcinogenic pathway of the benzo(a)pyrene-7,8-diol. Diethylstilbestrol and testosterone propionate resulted in small, but significant, decreases in metabolism of benzo(a)pyrene. When exposed in combination with 3-methyl-cholanthrene, testosterone propionate antagonized and diethylstilbestrol potentiated the metabolism of benzo(a)pyrene. 3-Methylcholanthrene, diethylstilbestrol, and combinations of 3-methylcholanthrene and diethylstilbestrol or testosterone propionate resulted in increased epoxide hydrolase activity as compared to controls. These results, carried out in a human hepatoma cell line, lend support to a concern for potentiated toxicity and carcinogenicity following exposure to complex chemical mixtures. 相似文献
885.
Hillary A. Hahm Margot M. Ip Kathleen Darcy Jennifer D. Black Wendy K. Shea Suzanne Forczek Masami Yoshimura Takami Oka 《In vitro cellular & developmental biology. Plant》1990,26(8):803-814
Summary A serum-free primary culture system is described which allows normal rat mammary epithelial cells (RMECs) embedded within
a reconstituted basement membrane to undergo extensive growth and functional differentiation as detected by synthesis and
secretion of the milk products casein and lipid. RMECs isolated from mammary glands of immature virgin rats were seeded within
an extracellular matrix preparation derived from the Engelbreth-Holm-Swarm sarcoma and cultured in a serum-free medium consisting
of Dulbecco's modified Eagle's medium-F12 containing insulin, prolactin, progesterone, hydrocortisone, epidermal growth factor,
bovine serum albumin, transferrin, and ascorbic acid. Casein synthesis and secretion were documented at the electron microscopic
level as well as by an enzyme-linked immunosorbent assay (ELISA) assay using a polyclonal antibody against total rat caseins.
Numerous secretory vesicles with casein micelles were noted near the apical surface of the RMECs, and secreted casein was
observed in the lumen. These ultrastructural data were confirmed by the ELISA assay which showed that microgram amounts of
casein per well were synthesized by the RMECs and that the amount of casein increased with time in culture. Using immunoblot
analysis it was demonstrated that the full complement of casein proteins was synthesized. In addition to casein protein, β-casein
mRNA levels were shown to increase with time. Synthesized lipid was detected at both the light and electron microscopic levels.
Phase contrast photomicrographs demonstrated extensive intracellular lipid accumulation within the ductal and lobuloalveolarlike
colonies, and at the electron micrograph level, lipid droplets were predominantly localized near the apical surface of the
RMECs. The lipid nature of these droplets was verified by oil red O staining. Results from this study demonstrate that RMECs
from immature virgin rats proliferate extensively and rapidly develop the capacity to synthesize and secrete casein and lipid
when grown within a reconstituted basement membrane under defined serum-free conditions. This unique system should thus serve
as an excellent model in which the regulation of mammary development and gene expression can be investigated.
This work was supported by grants CA 33240 and CA 35641 and by core grant CA 24538 from the National Institutes of Health,
Bethesda, MD. 相似文献
886.
Mitsuru Iwata Shoko Iwata Mark A. Everett Bryan B. Fuller 《In vitro cellular & developmental biology. Plant》1990,26(6):554-560
Summary A human foreskin organ culture system has been developed to study the response of human skin to hormonal stimulation. Foreskins
are maintained in culture on floating plastic supports which allows the epidermal surface to be exposed to air while the dermis
is bathed in nutrient medium. Both black and white human foreskins can be maintained in organ culture for at least 1 wk with
no change in the tissue structure or cell viability as determined by histochemical staining and by dopa reaction staining.
Tyrosinase activity in both black and white human foreskin cultures decays markedly during the first 2 d of culture to a new
steady state level which remains stable throughout the culture period. Both black and white foreskin cultures consistently
demonstrate 2- to 10-fold increases in tyrosinase activity when treated with theophylline (1 mM). Approximately 90% of all skin cultures examined showed an increase in enzyme activity when treated with this phosphodiesterase
inhibitor. Dibutyryl cAMP (0.1 mM) and [Nle4, D-phe7]-alpha MSH (10−8
M), were also found to markedly stimulate tyrosinase activity in some skin cultures, whereas alpha-MSH and prostaglandin E1 produced only an inconsistent and small increase in the activity of the enzyme. Histamine (1 μM), vitamin D3 (1 μM), and retinoic acid (1μM) failed to stimulate tyrosinase activity in either white or black foreskin cultures. This hormone-responsive organ culture
system can be utilized to characterize the molecular processes responsible for the regulation of tyrosinase and pigmentation
in human skin.
This work was supported by a research contract from the Oklahoma Center for the Advancement of Science and Technology (OCAST)
and by a research grant from the Presbyterian Health Foundation. 相似文献
887.
M. Moyer F. Bullrich J. B. Sheffield 《In vitro cellular & developmental biology. Plant》1990,26(11):1073-1078
Summary When embryonic retina is dissociated into a single cell suspension and maintained in stationary culture, a population of flat
cells is found on the culture dish. We have carried out a morphologic and immunologic study of the emergence of this population
in vitro. Ten- and fourteen-day-old chick embryo retinas were dissociated with trypsin, seeded on glass cover slips for various
times, and prepared for scanning electron microscopy (SEM) and immunofluorescence (IF) for Vimentin, an intermediate filament
protein. SEM indicates that the characteristic flat cell morphology is initiated in some cells in as little as 30 min after
the start of the culture. Not all of the cells that attach flatten. As incubation proceeds, small clusters of cells that had
formed in suspension attach to the substrate, and flat cells emerge from them. The flattened cells are positive for Vimentin
by IF within 10 min of attachment. The percent of fluorescent cells found on the substrate is constant during the time in
culture. This suggests that flat cells do not attach first, followed by neural cells, but that the neural cells and flat cells
attach to the dish at the same rate. When aggregates that had formed in suspension attach to the substrate, they are anchored
by flat cells that migrate out of the aggregate. Since Vimentin appears in the cultured cells within 10 min, it is unlikely
that it has been newly synthesized. Thus, the same cells that contained Vimentin in the retina now express it as flat cells.
this supports the hypothesis that flat cells derive from the same cells in the retina that give rise to Müller cells. We have
also observed the emergence of a population of cells with short (0.5μm) microvilli that appear within 8 h of culture. They
seem to be a distinct subpopulation of the cells on the upper portion of attached clusters.
This research was supported in part by grant EY-04892 and RR-0715 from the National Institutes of Health, Bethesda, MD, and
a grant from the W.W. Smith Foundations 相似文献
888.
Teresa L. Johnson Mary Pat Moyer 《In vitro cellular & developmental biology. Plant》1990,26(11):1095-1100
Summary Normal human colon mucosa cells and cells obtained from histologically normal tissues near that cancer were fused with human
colon cancer cells. Resultant hybrid populations of normal and malignant cell fusions behaved as nonmalignant cells in culture,
were unable to grow in soft agar, did not express tumor-associated antigens, and were nontumorigenic in nude mice. Autofusion
of the cancer cell population led to a phenotype intermediate between normal and malignant cells. That is, the cultures had
a much lower plating efficiency in soft agar, and the tumors had a longer latency and slower growth rate in nude mice. This
is the first cell culture system to demonstrate that normal epithelial cells can suppress malignancy of their autologous cancer
cells, and is a prelude to more extensive studies of genetic events involved in malignant conversion of human colonic epithelium.
This study was supported by The University of Texas Health Science Center at San Antonio Center for Human Cell Biotechnology
and a graduate student stipend (T. J.) from the Department of Cellular and Structural Biology. 相似文献
889.
Graham W. Burton Keith U. Ingold Kevin H. Cheeseman Trevor F. Slater 《Free radical research》1990,11(1):99-107
-Tocopherol, a superior chain-breaking, peroxyl radical-trapping antioxidant and the most active component of vitamin E, is elevated in liver tumor cells, contributing to their greater resistance towards lipid peroxidation compared to cells from normal tissues. Also, in regenerating rat liver the level of vitamin E has been found to fluctuate in phase with the rate of cell division. In order to study the biokinetcis and mechanisms of the distribution of vitamin E in organs and within tissues of animals, deuterated forms of -tocopherol have been synthesized and their uptake into blood and tissues has been measured by gas chromatography-mass spectrometry. Measurement of the competitive uptake from a mixture of the RRR-and SRR--tocopherol stereoisomers labelled with different amounts of deuterium shows that the liver exerts a strong preference for secretion of the natural (RRR) stereoisomer into the plasma. It is suggested that a tocopherol-binding protein plays a key role in this process. 相似文献
890.
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