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91.
The growth plate chondrocyte plays a central role in growth plate function. The purpose of this study was to characterize the respiratory and calcium transport properties of isolated mammalian growth plate chondrocytes and mitochondria obtained from these cells and to quantitate the mitochondrial weight and volume fraction in each zone of the growth plate. A new method was developed for isolation of mitochondria from chondrocyte suspensions. Isolated chondrocyte mitochondria demonstrated an eightfold increase in oxygen consumption in response to calcium and a two- to threefold increase in oxygen consumption in response to adenosine diphosphate. Similar responses were observed in chondrocytes treated with digitonin. The mitochondrial protein content of the growth plate and hyaline cartilage chondrocytes is significantly less than hepatocytes. Conversely, the chondrocyte mitochondrial cytochrome aa3 content is similar to mitochondria from a wide variety of sources. A zonal analysis of the growth plate demonstrates an increase in the mitochondrial weight (protein) fraction from the reserve to the hypertrophic zone whereas the mitochondrial volume fraction decreases from the reserve to the hypertrophic zone. The findings of this study emphasize the dependence of chondrocytes on glycolysis as a prime energy source and support the concept that chondrocyte mitochondria have become specialized in the process of matrix calcification.  相似文献   
92.
An attempt was made to concentrate plasma membranes of homogenized chondrocytes isolated by collagenase digestion of rachitic rat epiphyseal growth plate cartilage. This study reports the characterization of enzymes in the plasma membrane of isolated chondrocytes and their comparison with extracellular matrix vesicle components. The plasma membraneenriched fractions that were obtained showed a sevenfold increase in 5′-nucleotidase and a 15-fold increase in alkaline phosphatase, both of which are regarded as plasma membrane markers. SDS-polyacrylamide gel electrophoretic profiles of proteins extracted from membrane fractions contained several major protein bands also seen in isolated matrix vesicles. These studies indicate the usefulness of concentrating plasma membrane components from isolated chondrocytes, after the chondrocytes have been enzymatically freed from investing matrix and other stromal components by collagenase.  相似文献   
93.
94.
Articular chondrocytes have been thought to reside in a homogenous matrix. The physical characteristics of the intercellular matrix of articular cartilage are not well characterized, especially at a nanoscopic scale. The present work tested the hypothesis that the nanomechanical properties of the intercellular matrices of articular cartilage in both the articulating surface and various cellular zones are non-homogeneous. Nanoindentation by atomic force microscopy was applied to the geometric center of the medial, lateral and groove regions of the superficial zone of the rabbit proximal radius cartilage, and then the intercellular matrices of chondrocytes from the superficial to calcifying zones in 40 μm increments. The elastic modulus of the articular surface of the medial condyle (1.46±0.11 MPa) was significantly higher than the lateral condyle (1.18±0.10 MPa), and the groove (0.96±0.07 MPa). There is a significant gradient increase in Young’s moduli from the superficial zone (0.52±0.05 MPa) to calcifying zone (1.69±0.12 MPa). Thus, the nanomechanical properties of the intercellular matrices of the articulating surface are region-specific and likely related to articular function. Heterogeneous biophysical properties of intercellular matrices along the depth from the superficial to calcifying zones suggest that chondrocytes likely reside in a heterogeneous matrix.  相似文献   
95.
Insulin-like growth factor-I (IGF-I) is an important anabolic growth factor in the maintenance of articular cartilage phenotypic expression. Chondrocyte morphology is also tightly linked to phenotype. The small G-protein Cdc42 plays a key role in regulation of cell morphology and phenotypic expression in several cell types and, we show here, in articular chondrocytes. The purpose of these studies was to investigate possible links between the intracellular signaling pathways of IGF-I and Cdc42 in articular chondrocytes. Treatment of chondrocytes with IGF-I resulted in a rapid and sustained decrease in the activation state (decreased GTP-bound) of Cdc42. Nucleotide exchange and hydrolysis experiments suggest that the decreased activation occurs through increased hydrolysis. Transient expression of dominant-negative Cdc42(T17N) allowed for enhanced expression of normal chondrocyte phenotype as determined by increased mRNA expression of collagen type II (Coll II) with decreased matrix metalloproteinase-3 (MMP-3) expression. The results of these studies suggest a novel link between IGF-I and Cdc42 signaling pathways. Further, an additional mechanism for the regulation of chondrocyte phenotype is defined through the IGF-I induced down-regulation of Cdc42 activation.  相似文献   
96.
Previous work has shown that interleukin 1 (IL-1) increases the activity of acid extruders in articular chondrocytes, while the H+-adenosine triphosphatase (ATPase) inhibitor bafilomycin can prevent aggrecanase-mediated cartilage degradation. The H+ transport induced by IL-1 may therefore be required for proteinase activity. In the present study, the effects of hexosamines and fish oils on H+-ATPase activity have been characterised for isolated bovine articular chondrocytes. Cells isolated in the presence of IL-1 were acidified, and the fraction of acid extrusion mediated by Na+–H+ exchange and an H+-ATPase were determined using specific inhibitors. Exposure to IL-1 significantly enhanced both components of acid extrusion. Co-incubation with glucosamine or mannosamine attenuated the H+-ATPase fraction of efflux. The addition of glucosamine at 9 h after exposure to IL-1—when H+-ATPase activation is already apparent—was also able to abolish H+-ATPase activity, implying that hexosamines do not exert effects at the level of protein synthesis. Co-incubation with the glucose transport inhibitor phloretin elicited similar effects to the hexosamines, suggesting that modulation of adenosine triphosphate levels may underlie their effects on H+-ATPase function. The omega-3 fish oil linolenic acid but not the omega-6 fish oil linoleic acid reduced H+-ATPase activity to levels seen in IL-1-untreated cells, although total efflux remained elevated, as a result of an enhanced H+ leak. These observations support a model whereby IL-1 stimulates an H+-ATPase-dependent system, possibly involved in aggrecanase activation, which appears to be one of the target mechanisms interrupted by dietary supplements reported to have symptom-modifying effects on osteoarthritis.  相似文献   
97.
High-serum media have been shown to produce significant improvement in the properties of tissue-engineered articular cartilage when applied in combination with dynamic deformational loading. To mitigate concerns regarding the culture variability introduced by serum, we examined the interplay between low-serum/ITS-supplemented media and dynamic deformational loading. Our results show that low serum/ITS supplementation does not support the same level of tissue formation as compared to high serum controls. In free-swelling culture, using a combination of ITS with concentrations of FBS above 2% negated the beneficial effects of ITS. Although there were beneficial effects with loading and 0.2%FBS + ITS, these constructs significantly underperformed relative to 20%FBS constructs. At 2%FBS + ITS, the free-swelling construct stiffness and composition approached or exceeded that of 20%FBS constructs. With dynamic loading, the properties of 2%FBS + ITS constructs were significantly lower than free-swelling controls and 20%FBS constructs by day 42. By priming the chondrocytes in 20%FBS prior to exposure to low-serum/ITS media, we observed that low-serum/ITS media produced significant enhancement in tissue properties compared to constructs grown continuously in 20%FBS.  相似文献   
98.
99.
Objectives: The aim of the present study was to investigate the factors that contribute to the progression of synovial chondromatosis in the temporomandibular joint (TMJ).

Methods: The authors investigated the expression of CD105 and CD90 in specimens from 17 patients with synovial chondromatosis in the TMJ, using immunohistochemical staining, and expression of CD105 and CD90 in cartilaginous nodules was scored semiquantitatively.

Results: The expression of CD105 and CD90 was found in almost all the cases. In particular, the expression of CD90 in cartilaginous nodules significantly decreased with the progression of synovial chondromatosis.

Discussion: The factors that determine progression of synovial chondromatosis are not fully understood. The results of this study suggest that CD90 may play an important role in the progression of synovial chondromatosis in the TMJ.  相似文献   

100.
The purpose of this study was to elucidate the expression of proto-oncogene Bcl-2 (anti-apoptotic) and Bax (pro-apoptotic) in fibrocartilage of the disc and hyaline cartilage of the condyle in the rabbit craniomandibular joint (CMJ). Ten New Zealand white rabbit heads were used. Sections were processed by the immunohistochemical techniques using mouse anti-Bcl-2 and anti-Bax antibodies. Intensity levels of immunostaining in condylar cartilage were quantified by a computer-image system. Immunoreactivity for Bcl-2 was mainly observed in the cytoplasm of the reserve cell and chondrocytic cell layers. A mild heterogeneous Bax expression was detected in the cytoplasm of chondrocytes of the upper hypertrophic layer and a few cells of the chondrocytic layer. The cytoplasm of chondrocytes in the disc exhibited a high intensity for Bcl-2, while Bax activity was only sporadically observed. We have shown that Bcl-2 and Bax proteins are present in CMJ cartilage and their expression patterns suggest that these oncoproteins are involved in chondrocyte survival or death via apoptotic pathways.  相似文献   
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