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91.
Summary Human umbilical vein endothelial cells (HUV-EC) grew rapidly in vitro in medium supplemented with epidermal growth factor,
fetal bovine serum (FBS) and human diploid fibroblast-conditioned medium. The effect of FBS could be replaced partially by
bovine serum albumin, cholesterol, and vitamin E, and completely by further addition of serum dialysate or refeeding every
other day. Among these components, fibroblast-conditioned medium is essential for HUV-EC growth. The HUV-EC were cultured
serially for over 50 population doublings in the 10% FBS containing fibroblast-conditioned medium and for over 40 population
doublings in the serum-free medium. Mitogenic factor(s) present in the medium conditioned by fibroblasts may be related to
endothelial cell growth factor and play an important role angiogenesis and regeneration of vascular endothelium in vitro. 相似文献
92.
Steinar Solberg Tor Larsen Leif Jørgensen 《In vitro cellular & developmental biology. Plant》1985,21(11):612-616
Summary This study examined whether nonconfluent endothelial cell cultures reacted differently than confluent ones toward thrombin-stimulated
platelets or a heparinized salt solution. The adherence to the endothelial cell cultures of51Cr-labeled human platelets stimulated at different thrombin concentrations was studied. There was significantly higher adherence
of stimulated platelets to nonconfluent cultures compared with confluent ones. This was confirmed by scanning electron microscopy,
which also revealed a tendency for the platelets to adhere at the cell periphery. Electron microscopy also showed that thrombin-stimulated
platelets induced endothelial cell contraction. Part of the peripheral endothelial cell surface toward the bottom of the culture
dish was inverted, facing the lumen of the dish. This phenomenon was particularly seen in nonconfluent cultures. When51Cr-labeled endothelial cultures were incubated with a mildly injurious fluid as heparinized sodium acetate and 20% serum,
at 20° C for 30 min, the nonconfluent cultures showed significantly more cell detachment and release of51Cr than the confluent ones. We conclude that under the conditions of the present experiments there are differences in the
reactivity of confluent and nonconfluent endothelial cell cultures. These differences probably reflect biological dissimilarities.
In experiments where properties of cultured endothelium are studied, care should be taken that the degree of confluency is
standardized. 相似文献
93.
Degradation of extracellular matrix by the trophoblastic cells of first-trimester human placentas 总被引:6,自引:0,他引:6
S J Fisher M S Leitch M S Kantor C B Basbaum R H Kramer 《Journal of cellular biochemistry》1985,27(1):31-41
First-trimester human placental villi were cultured on 3H-leucine-labeled extracellular matrices isolated from the PF HR9 and PYS-2 cell lines. Both cell lines produced an extracellular matrix that contained basement membrane-specific macromolecules, including type IV collagen, laminin and proteoglycan. Both matrices promoted outgrowth of cells from the villi which, according to morphological criteria, were identified as cytotrophoblastic cells. As the cells migrated from the attachment site, they caused a marked focal dissolution of the matrix which was accompanied by a concomitant release of 3H-labeled material into the media. Approximately half of this material chromatographed near the inclusion volume of Sephadex G-50, indicating that the labeled matrix components had been degraded. This phenomenon was dependent on the age of the placenta. Second-trimester placental villi also adhered to the matrix, but no areas of dissolution were formed and no significant amounts of radioactivity were released into the medium. These results suggest that culture of first-trimester human placental villi on extracellular matrices may be useful for the study of some of the early embryonic events leading to human implantation, during which the trophoblastic cells erode the uterine epithelium. 相似文献
94.
95.
Effects of fructose on human fibroblast metabolism: the application of DNA measurements as a basis for interpretation 总被引:2,自引:0,他引:2
Robert Z. Eanes 《In vitro cellular & developmental biology. Plant》1985,21(6):328-332
Summary A fluorometric procedure for measuring DNA was used to study growth and metabolic responses of eight cell strains of human
foreskin fibroblasts. In preliminary studies this procedure gave more precise specific activity changes inN-acetyl-β-d-glucosaminidase (NAG) than did a protein activity basis, when changes in this enzyme's specific activity were investigated
as a function of experimental cell manipulation. When fibroblast growth in eight cell strains was compared in 134 mM
d-fructose vs. 13.4 mM glucose-supplemented minimum essential media, a significant increase in cellular DNA (50%) and protein (45%) occurred over
an 11-d period. No significant differences in media pH change, lactate production, or carbohydrate uptake occurred on a DNA
basis when cell metabolism was compared over the last 24 h of culture in the two media. Cells grown in fructose-containing
media tended to show a reduction in NAG specific activity when compared with those grown in glucose-containing media. 相似文献
96.
John E. Gould James W. Overstreet Frederick W. Hanson 《Molecular reproduction and development》1985,12(1):47-54
The functional capacity for sperm interaction with the human zona pellucida and zona-free hamster oocyte was tested after human spermatozoa were capacitated by passage through a column of human cervical mucus. The results were compared with those obtained when spermatozoa from an aliquot of the same semen sample were capacitated by the standard laboratory methods involving sequential washing by dilution and centrifugation of the semen. Washed-capacitated sperm suspensions were more successful than mucus-capacitated sperm in attaching to the zona-free hamster oocyte and in fusing with its oolemma. However, the ability of mucus-capacitated sperm to penetrate the human zona pellucida was equal to washed capacitated sperm. These experiments demonstrate an approach that may be useful in comparative studies of human sperm capacitation in vivo and in vitro. 相似文献
97.
Age-dependent metabolic changes in cultured human fibroblasts 总被引:1,自引:0,他引:1
A. L. Muggleton-Harris R. Defuria 《In vitro cellular & developmental biology. Plant》1985,21(5):271-276
Summary The effects of metabolic poisons on the ATP content of cultured human skin fibroblasts at selected in vitro and in vivo ages
were studied. Potassium cyanide, iodacetemide, and Arsenate were used to inhibit ATP restoration by glycolysis and oxidative
phosphorylation. Cells treated with these metabolic poisons showed an age-dependent change in their ATP content. The decrease
in cellular ATP content after exposure to these drugs was taken as an estimate of ATP turnover. It was found that there was
a decrease in the ATP turnover with increasing population doubling level (i.e. in vitro age), and cells cultured from a 68-yr-old
donor had a lower ATP turnover than those cultured from a neonatal donor. This decreased ATP turnover correlates with a previous
finding of a decreased ability of “older” cells to be stimulated to migrate in culture and suggests that there is a metabolic
component to this age-related functional deficiency.
This work was supported by National Institutes of Health grants 2, RO1 EY02523 and 1 RO1 1, AGO 1212 awarded to A.L. Muggleton-Harris. 相似文献
98.
A. Shteyer D. Gazit I. Binderman I. A. Bab 《In vitro cellular & developmental biology. Plant》1987,23(1):15-20
Summary Cells of the dental papilla are capable of odontoblastic, fibroblastic, and endothelial differentiation and formation of dentin
and the dental pulp. In the present study dental papilla cells, obtained from human tooth buds (HDP cells), were cultured
in vitro through 3 to 7 passages. After exposure to prostaglandin E2 there was a marked decrease in intracellular cyclic AMP (cAMP) levels as compared to hormone-free controls. Parathyroid hormone
and calcitonin had stimulatory effects with 1 and 2 log increases in cAMP, respectively. The HDP cells showed moderate activity
of alkaline phosphatase, 1 log higher than that of hamster kidney fibroblasts (BHK 13) and 1 log lower than that of osteoblastic
osteosarcoma cells (ROS 17/2). When cultured for 4 or 8 wk in diffusion chambers (DC) implanted in athymic mice, many of the
HDP cells underwent odontoblastic morphodifferentiation with very long, single processes extending into the matrix. This matrix
contained banded and unbanded collagen fibers. Neither light nor electron microscopy of the DC content revealed mineral deposits.
These results suggest that HDP cells have an intrinsic potential for partial odontoblastic differentiation; inductive signals
like those originating from odontogenic epithelium are probably essential for the completion of hard tissue formation. 相似文献
99.
Cytochromec oxidase was isolated from human hearts and separated by SDS gel electrophoresis. The identity of polypeptide bands with known subunits was demonstrated by immunoblotting with monospecific antisera to rat liver cytochromec oxidase subunits. The polarographically determined kinetics of cytochromec oxidation were similar to those reported for the bovine heart enzyme. 相似文献
100.
Biochemical significance of enhanced activity of fluorinated 1,25-dihydroxyvitamin D3 in human cultured cell lines 总被引:1,自引:0,他引:1
Several human cancer cells possess receptors for 1,25-dihydroxyvitamin D3[1,25-(OH)2D3]. In these cells 1,25-(OH)2D3 has a biphasic concentration-dependent regulatory effect on cell replication and specifically induces its own metabolism. We have studied the effects on these parameters of the native hormone together with those of two analogues fluorinated at the 24-carbon and of 1,24R,25-trihydroxyvitamin D3[1,24R,25-(OH)3D3]. The difluorinated analogue 24,24-difluoro-1,25-(OH)2D3[24,24-F2-1,25-(OH)2D3] is an approximately fivefold more potent inhibitor of cellular replication than the native hormone, while 1,24R,25-(OH)3D3 is about fivefold less potent. This enhanced potency of the fluorinated analogue parallels its enhanced potency in in vivo studies of its effects on calcium and mineral metabolism. However, although the analogue retains replication stimulatory activity, it is clearly no more potent than the native hormone in this activity: 1,24R,25-(OH)3D3 has no significant stimulatory activity. Exposure of the cells to 1,25-(OH)2D3 at 0.05 nM for 6 h increases the subsequent conversion of labelled hormone to aqueous phase soluble compounds by 6.7-fold. None of the other compounds had a similar effect at this concentration. At 10 nM all 1-hydroxylated compounds increased aqueous phase radioactivity about equally (13 to 17-fold); this effect is still specific since 25-OH D3 had no such effect even at 10 nM. Studies on the effects of the fluorinated analogues upon receptor binding of hormone in cell cytosols and uptake of hormone by intact cells clearly demonstrate that the enhanced activity of these analogues is not due to higher receptor affinity or more rapid access to intracellular receptor.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献