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71.
Endocrine cell cultures have potential in bioprocessing, for the production of biologically active hormones, and in tissue engineering, for the development of implantable artificial tissues for long-term restoration of endocrine function. To optimize such systems, it is necessary to develop a thorough understanding of how inherently present environmental stresses, such as nutrient depletion and metabolite accumulation, affect the cells. This work focuses on the effects of the metabolite ammonium on indicators of endocrine cell metabolism and on the processing, storage and secretion of regulated secretory proteins. Experiments were conducted on recombinant insulin-producing mouse pituitary AtT-20 cells and mouse insulinoma TC3 cells. Exposure for 24–48 hours to 6 mM of exogenous ammonium resulted in higher rates of glucose consumption by both AtT-20 and TC3 cells, while the formation of additional ammonium generally decreased relative to ammonium-free controls. When TC3 cells were discharged of their intracellular insulin stores, the presence of ammonium during a subsequent recharge completely inhibited addition of new insulin-related peptides to the stores, as we had observed previously for both cell lines. There was a correlation between insulin-related peptides stored in TC3 cells during recharging and the amount that could be released upon secretagogue stimulation. Using a combination of radioimmunoassay and high performance liquid chromatography, we found that intracellular insulin and insulin-related peptides changed in the same fashion. Intracellular mechanisms that may be producing the observed results are discussed.Abbreviations IRP
insulin-related peptides
- HPLC
high performance liquid chromatography
- DAMP
3-(2,4-dinitroanilino)-3 amino-N-methyldipropylamine 相似文献
72.
Summary The organ culture of the mammary gland of lactating mice was used to examine the response of the differentiated gland to lactogenic
stimuli, insulin, cortisol, and prolactin. Time course studies showed that casein synthesis in cultured tissue decreased rapidly
during the first 2 d despite the presence of the three hormones, but on the 3rd d tissue cultured with either insulin and
prolactin or all three hormones regained the ability to synthesize milk proteins, casein, and α-lactalbumin: a greater increase
occurred in the three hormone system. The delayed addition of prolactin on Day 2 to the culture system containing insulin
and cortisol also stimulated casein synthesis. The addition of cytarabine, which inhibited insulin-dependent cell proliferation
in cultured explants, did not block the rebound of milk protein synthesis. The results indicate that in the presence of insulin,
cortisol, and prolactin mammary epithelial cells in culture first lose and then regain the ability of synthesizing milk protein
without requiring the formation of new daughter cells. 相似文献
73.
Insulin and glucagon were labeled with iodine. The reaction products were analyzed by high-performance liquid chromatography. It is shown that the pH of the reaction medium has a large effect on the position and the degree of iodine substitution as well as on the oxidation of the Met-containing glucagon and, furthermore, that the molar ratio of iodine to polypeptide hormone used during the labeling procedure affects not only the amount of iodine incorporated but also the distribution of iodinated products. The results show that certain iodinated derivatives are separated from each other and from the respective unlabeled polypeptide and thus can be obtained in a pure state. 相似文献
74.
James A. McAteer Orion D. Hegre 《In vitro cellular & developmental biology. Plant》1978,14(9):795-803
Summary A method of perfusion organ culture is described in which explants cultured at the airmedium interface are bathed by a continuous
flow of nutrient medium. Morphological studies on the fetal rat lung indicate that explant development in this system is comparable
to that obtained using standard organ-culture dishes. Medium supply is easily manipulated and continuous sampling of the effluent
stream is possible without disturbing the immediate explant environment. The basic design facilitates secretory-response studies
on cultured organ explants as demonstrated by a study of glucose-stimulated insulin release by the neonatal rat pancreas.
This work was supported by U. S. Public Health Service Training Grant No. GM 00114. 相似文献
75.
Summary Insulin release and membrane potential fluctuations in response to increased extracellular potassium [K+]
o
have been measured in single perifused islets of Langerhans from normal mice. An increase in [K+]
o
from 5mm to 50mm induced a transient insulin release with a peak at about 1 min. The peak value was [K+]
o
-dependent but the half-timet
1/2 for the decline was constant at nearly 1 min. 2.5mm cobalt completely inhibited the potassium-induced stimulation of insulin release. The insulin release elicited by 28 and 50mm [K+]
o
was similar in terms of peak, total release and half-time from maximum release. Stepwise increase in [K+]
o
from 10 to 28 to 50mm resulted in a normal response to 28mm but no peak of release after the 28 to 50mm increase. The results indicate good correlation between excess voltage noise, thought to reflect calcium channel activity, and insulin release evoked by changing extracellular potassium. 相似文献
76.
Effect of bombesin upon plasma somatostatin-like immunoreactivity, insulin and glucagon in normal and chemically sympathectomized dogs 总被引:2,自引:0,他引:2
The present study was designed to determine the effects of intravenously infused bombesin (10 ng/kg/min) upon basal and postprandial plasma somatostatin-like immunoreactivity (SLI), glucagon, insulin and triglyceride levels in normal (n = 12) and chemically sympathectomized (n = 11) dogs. Basal plasma SLI, glucagon and insulin levels rose significantly during the infusion of bombesin in the normal dogs, and this was not altered by chemical sympathectomy. Bombesin infusion enhanced the postprandial SLI response, while attenuating the postprandial glucagon response by 50% and the insulin response in the early postprandial phase of the meal. Sympathectomy did not significantly alter the basal levels of these polypeptides, but augmented the postprandial plasma SLI response during the first 90 min, and reduced the postprandial glucagon response during the infusion of bombesin. The postprandial insulin response was not affected by sympathectomy. In both normal and chemically sympathectomized dogs the rise in postprandial triglyceride levels was attenuated by bombesin infusion. 相似文献
77.
Hexokinase is present in the tissues in four isoenzymic forms. Cerebral tissue contains predominantly Type I hexokinase which
is believed to be insulin-insensitive. In cerebral tissue about 60 to 70% of the hexokinase is bound to the particulate fraction.
The changes in the distribution of hexokinase Type I and Type II together with the bound and free hexokinase have been studied
in control, diabetic and diabetic animals treated with insulin. The results indicate that the presence of insulin is essential
for the normal binding of the hexokinase to the particulate fraction. In heart tissue, Type II hexokinase bound to the pellet
shows a significant decrease in diabetes, which is reversed on insulin administration. 相似文献
78.
The immune responses to several antigens were compared in the I-A mutant mouse strain B6.C-H-2bm12 and the wild-type strain C57BL/6. With a lymph node cell proliferation assay, the response to two of these antigens, beef insulin and (TG)A-L, was demonstrated to be controlled by a gene in the I-Ab region. B6.C-H-2bm12 mice failed to respond to beef insulin, while their responses to (TG)A-L, DNP-OVA and PPD were comparable with those of the wild-type strain C57BL/6. Taken together with previous studies, these data suggest that the product of a single pleiotropic I-A gene, an la molecule, functions as a histocompatibility, la, and MLR antigen, as well as a necessary component for Ir gene function. Furthermore, the data reported here demonstrate that la molecules have multiple functional “Ir determinants,” one of which has been altered in the B6.C-H-2bm12 mutant. The B6.C-H-2bm12 mice, therefore, represent a powerful analytical tool for the understanding of the cellular and molecular basis for Ir gene control of the immune response. 相似文献
79.
A review of experimental studies of the effect of zinc nutrition on insulin metabolism is presented. In addition to a short
introduction to the synthesis, secretion, and action of insulin, the effects of zinc deficiency—specifically on glucose tolerance,
insulin secretion, insulin synthesis and storage, and on total insulin-like activity—are dealt with. The concentrations of
zinc and chromium in serum, pancreas, and liver are compared to those of zinc-deficient animals and pair-fed controls.
In contrast to pair-fed controls, zinc-deficient rats had unaltered proinsulin contents after glucose stimulation, but they
showed a diminished glucose tolerance, lowered serum insulin content, and an elevated total insulin-like activity. The serum
zinc concentration of the deficient animals was greatly reduced and did not change during glucose stimulation, whereas it
rose in the case of the pair-fed controls. The serum chromium concentration increased in both groups in response to glucose
stimulation. In the pancreas of the deficient animals, the zinc concentration was reduced 60% and it increased during the
glucose tolerance test. In the liver there were no significant differences. The chromium concentrations were elevated in both
the pancreas and liver of the zinc-deficient rats by 60 and 100%, respectively, and were not influenced by glucose injection.
These studies show clearly that nutritional zinc deficiency influences insulin metabolism and action. 相似文献
80.
Eight Billroth II resected patients and 8 normal controls were given two oral glucose loads, one ingested within 2 min, and the other ingested slowly over 80 min. In the Billroth II resected group, the integrated plasma GIP release was significantly higher after the fast than after the slow glucose ingestion. In this group the integrated plasma GIP release was also significantly higher than in the control group, but only after the fast glucose ingestion. These findings indicate that the rate of glucose delivery into the intestine may be of importance in the plasma GIP response to oral glucose. 相似文献