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21.
Y Gache F Landon H Touitou A Olomucki 《Biochemical and biophysical research communications》1984,124(3):877-881
Purified alpha-actinin from human platelets was digested with Ca2+-activated protease from muscle. The alpha subunit (Mr = 100 kDa) was degraded into a unique polypeptide b of slightly lower molecular mass. In fresh platelets, only the a subunit was detected by immunoblotting techniques, while in out-dated platelets, both a and b polypeptides were present. Since a similar conversion of a to b occurs in vitro as in whole platelets, it can be assumed that, in platelets, alpha-actinin is cleaved by the endogenous Ca2+-activated protease. 相似文献
22.
The sperm head of the plains mouse, Pseudomys australis, has three curved hooks projecting from its anterior margin. The two ventral hooks have previously been shown to consist largely of an extension of the subacrosomal material. To characterize further the structure and composition of the ventral hooks, we have examined their formation during spermiogenesis using transmission electron microscopy, silver staining, and actin localization with NBD-phallacidin. The ventral hooks develop as an extension of the perinuclear space and postacrosomal dense lamina on the anteroventral margin of the sperm head. Bundles of 6-nm-thick filaments appear in the core of each hook; these are probably actin filaments based on staining of the hooks with NBD-phallacidin. Just prior to spermiation, electron-dense material condenses in the core of the ventral hooks and concurrently in the perinuclear space in the remainder of the sperm head. The two ventral hooks thus appear to consist of a core of perinuclear material and actin filaments, which is enclosed by a continuation of the postacrosomal dense lamina. 相似文献
23.
C. Méjean M. Boyer J. P. Labbé J. Derancourt Y. Benyamin C. Roustan 《Bioscience reports》1986,6(5):493-499
The interaction of two different anti-actin antibody populations with the myosin subfragment 1-F-actin rigor complex has been studied. In contrast with the 1–7 sequence, the 18–28 sequence appears to be strongly implicated in the contact area of the myosin head on the actin polypeptide chain. 相似文献
24.
T D Pollard 《Journal of cellular biochemistry》1986,31(2):87-95
25.
Human arterial smooth muscle cells in culture: Inverse relationship between proliferation and expression of contractile proteins 总被引:5,自引:0,他引:5
Gunnar Fager Göran K. Hansson Allen M. Gown David M. Larson Omar Skalli Göran Bondjers 《In vitro cellular & developmental biology. Plant》1989,25(6):511-520
Summary Human arterial smooth muscle cells (hASMC) from explants of the inner media of uterine arteries were studied in secondary
culture. We had previously found that these cells depend on exogenous platelet-derived growth factor (PDGF) for proliferation
in vitro. Deprivation of the serum mitogen(s) by culture in plasma-derived serum or bovine serum albumin (BSA) caused a true
growth arrest that was reversible upon reexposure to the mitogen(s). When added to serum-containing medium, heparin caused
a reversible growth arrest which could be competed for by increasing concentrations of serum. In the current study we used
a set of smooth muscle-specific actin and myosin, antibodies to study the expression of contractile proteins in stress fibers
under indirect immunofluorescence on hASMC in culture. Even in sparse culture, grwoth-arrested hASMC expressed stress fibers
containing these actin and myosin epitopes. This was true irrespective of whether growth arrest was achieved by culture in
media containing only BSA or a combination of heparin and whole blood serum. hASMC proliferating in whole blood serum in sparse
culture did not express such strees fibers, as judged by immunofluorescent staining. This was true also for cells that were
restimulated to proliferate in serum after a growth arrest. Utilizing a monoclonal antibody against a nuclear antigen expressed
in proliferating human cells, we were able to demonstrate an inverse relationship between the expression of this antigen and
the SMC-specific contractile proteins, respectively. Under these culture conditions, the reversible transition between defifferentiated
and differentiated hASMC was almost complete and terminated about 1 wk after the change in culture condition. We conclude
that hASMC in vitro respond, to exogenous PDGF by proliferation and dedifferetiation as a single population of cells. We also
conclude that this modulation is reversible, because the cells become uniformly quiescent and differentiated when the mitogenic
stimulus is blocked or removed.
This study was supported by grants from the Swedish Medical Research Council (Project no. 4531 and 6816), the Swedish Association
against Heart and Chest Diseases, the King Gustaf V and Queen Victoria Foundation, the National Institutes of Health, Bethesda,
MD (grant HL 29873) and the Swedish National Board for Laboratory Animals. 相似文献
26.
Summary Alpha-smooth muscle actin is currently considered a marker of smooth muscle cell differentiation. However, during various
physiologic and pathologic conditions, it can be expressed, sometimes only transiently, in a variety of other cell types,
such as cardiac and skeletal muscle cells, as well as in nonmuscle cells. In this report, the expression of actin mRNAs in
cultured rat capillary endothelial cells (RFCs) and aortic smooth muscle cells (SMCs) has been studied by Northern hybridization
in two-dimensional cultures seeded on individual extracellular matrix proteins and in three-dimensional type I collagen gels.
In two-dimensional cultures, in addition to cytoplasmic actin mRNAs which are normally found in endothelial cell populations,
RFCs expressed α-smooth muscle (SM) actin mRNA at low levels. α-SM actin mRNA expression is dramatically enhanced by TGF-β1. In addition, double immunofluorescence staining with anti-vWF and anti-α-SM-1 (a monoclonal antibody to α-SM actin) shows
that RFCs co-express the two proteins. In three dimensional cultures, RFCs still expressed vWF, but lost staining for α-SM
actin, whereas α-SM actin mRNA became barely detectable. In contrast to two-dimensional cultures, the addition of TGF-β1 to the culture media did not enhance α-SM actin mRNA in three-dimensional cultures, whereas it induced rapid capillary tube
formation. Actin mRNA expression was modulated in SMCs by extracellular matrix components and TGF-β1 with a pattern very different from that of RFCs. Namely, the comparison of RFCs with other cell types such as bovine aortic
endothelial cells shows that co-expression of endothelial and smooth muscle cell markers is very unique to RFCs and occurs
only in particular culture conditions. This could be related to the capacity of these microvascular endothelial cells to modulate
their phenotype in physiologic and pathologic conditions, particularly during angiogenesis, and could reflect different embryologic
origins for endothelial cell populations.
Supported by a Post-Doctoral Fellowship from the Swiss National Science Foundation (OK) and grant HL-RO1-28373 (JAM) from
the Department of Human Services, Public Health Service, Washington, D.C. 相似文献
27.
Mark Phillippe Trevania Saunders Shrikar Bangalore 《In vitro cellular & developmental biology. Plant》1990,26(4):369-378
Summary The following studies were undertaken to develop a cultured uterine myocyte model which would allow further clarification
of the adrenergic signal transduction mechanisms utilized by these myocytes. After mechanical removal of the endometrium,
rabbit uterine myoctes were isolated by an overnight enzymatic disaggregation using collagenase and DNase I. The isolated
myocytes were maintained in culture in 75-cm2 flasks containing Waymouth's MB 751/1 medium-10% fetal bovine serum along with 10−8
M estradiol, penicillin, streptomycin, and Fungizone. The phase contrast and electron micrographic appearance of these cells
was consistent with that previously reported for smooth muscle myocytes in culture. Immunocytochemical studies utilizing monoclonal
anti-alpha-smooth muscle actin antibodies confirmed the presence of smooth muscle actin in these cultured myocytes. Western
blot studies similarly confirmed the presence of alpha-smooth muscle actin in rabbit myometrial tissue and the cultured myocytes,
both the primary and F1 generation. After prelabeling the myocytes with [3H]inositol, adrenergic stimulation experiments demonstrated alpha-1 receptor mediated stimulation of inositol phosphates.
Beta receptor stimulation experiments confirmed cAMP production in these cultured myocytes, and the ability of clonidine,
an alpha-2 agonist, to inhibit forskolin stimulated cAMP production confirmed the presence of functional alpha-2 adrenergic
receptors in these myocytes. In conclusion, these cultured rabbit uterine myocytes have provided an in vitro model which can
be utilized to further clarify the adrenergic receptor signal transduction mechanisms in genital tract smooth muscle.
This research was supported by grant HD-22063 from the National Institutes of Health, Bethesda, MD. 相似文献
28.
MARIA APARECIDA VISCONTI ANA MARIA DE LAURO CASTRUCCI 《Pigment cell & melanoma research》1990,3(3):132-140
Norepinephrine (NE) and phenylephrine (Phe) were employed to study the ionic requirements for alpha adrenoceptor activation in the teleost Poecilia reticulata melanophores. As expected the beta adrenoceptor blocker, propranolol, increased the sensitivity of the preparation to NE (5.8 times), and was therefore employed in all the experimental procedures. Neither cocaine (a neuronal uptake blocker) nor dexamethasone (an extraneuronal uptake blocker) enhanced the sensitivity of the preparation to NE, suggesting that these inactivating mechanisms would not play a role in P. reticulata pigmentary system. However, in the absence of calcium, the dose-response curve (DRC) to NE was displaced to the left about 3.5 times, whereas the DRC to Phe was not affected. These results indicate that a neuronal uptake is active, but was not demonstrated by the classical pharmacological tools, probably due to an assymmetric display of the nervous endings. The DRC to NE was rightward displaced (14.1 times) in the presence of the calcium channel blocker Verapamil, whereas the DRC to Phe was not affected. These data suggest that P. reticulata melanophores possess a mixed population of alpha1 and alpha2 adrenoceptors, the activation of the latter eliciting an extracellular calcium influx. In sodium-free saline, the DRC to NE was rightward shifted (6.6 times) and the response to Phe was impaired in such a way that the maximal response was not achieved. The DRC to both NE and Phe were rightward displaced (7.9 and 2.7 times respectively) in the presence of the sodium channel blocker tetrodotoxin (TTX) 10?7M. In potassium-free saline, the melanophore sensitivity to Phe was increased, whereas the responses to NE were not affected, suggesting a differential sensitivity of the two alpha adrenoceptor subtypes to the resulting membrane hyperpolarization. Based on the literature and on our data we propose that P. reticulata melanophores possess a mixed population of alpha1 and alpha2 receptors. The activation of both subtypes of alpha adrenoceptors elicits a Na+ ion influx through TTX-sensitive sodium channels. The stimulation of alpha2 adrenoceptors also requires an extracellular calcium influx, through the opening of slow calcium channels. 相似文献
29.
30.
Shinichiro Nakamura Wijit Kiatipattanasakul Hiroyuki Nakayama Fumiko Ono Ippei Sakakibara Yasuhiro Yoshikawa Naoaki Goto Kunio Doi 《Journal of medical primatology》1996,25(4):294-300
Abstract: In this study, we immunohistochemically examined the several constituents of senile plaques (SPs) and cerebral amyloid angiopathy (CAA) in aged cynomolgus monkeys. Apolipoprotein E (apoE) deposited in all mature plaques and CAA, and in half of the diffuse plaques. Alpha-1-antichymotripsin (αACT) deposited in half of the mature plaques and in one third of the CAA. Amyloid precursor protein (APP), ubiquitin (Ub), and microtubule-associated protein-2 (MAP-2) accumulated in the swollen neurites of mature plaques. Glial fibrillary acidic protein (GFAP) was detected in the astrocytes and their processes surrounding the mature plaques. Tau was detected in neither the SPs nor CAA. Therefore, mature plaques involved extracellular Aβ, apoE, and αACT, and also astrocytes and swollen neurites. However, diffuse plaques involved only extracellular Aβ and apoE. Since these features, except for tau, were consistent with those in humans, this animal model will be useful for studying the pathogenesis of cerebral amyloid deposition. 相似文献