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51.
Preparation of P32-phosphanilic acid-bovine serum albumin tracer 总被引:1,自引:0,他引:1
52.
Podoplanin is expressed by a sub-population of human foetal rib and knee joint rudiment chondrocytes
The aim of this study was to determine if podoplanin was expressed by rudiment chondrocytes in human foetal cartilages. Podoplanin was immunolocalised in first trimester human foetal rib and knee joint rudiments to a sub-population of chondrocytes deep in the rib rudiments, tibial and femoral growth plates and cells associated with the cartilage canals of the foetal knee joint rudiments. Lymphatic vessels in the loose stromal tissues surrounding the developing rudiments were also demonstrated on the same histology slides using antipodoplanin (MAb D2-40) and anti-LYVE-1 and differentiated from CD-31 positive blood vessels confirming the discriminative capability of the antibody preparations used. The D2-40 positive rib and knee rudiment chondrocytes were not stained with antibodies to LYVE-1, CD-31 or CD-34 however perlecan was a prominent pericellular proteoglycan around these cells confirming their chondrogenic phenotype. Discernable differences were evident between the surface and deep rudiment chondrocytes in terms of their antigen reactivities detected with MAb D2-40 or antiperlecan antibodies. Binding of the cytoplasmic tail of PDPN to the ERM proteins ezrin, radixin and moeisin may result in changes in cytoskeletal organisation which alter the phenotype of this central population of rudiment cells. This may contribute to morphological changes in the rudiment cartilages which lead to establishment of the primary ossification centres and is consistent with their roles as transient developmental scaffolds during tissue development. 相似文献
53.
Globally, 40 million people live with the chronic effects of lymphatic filariasis (LF), making it the second leading cause of disability in the world. Despite this, there is limited research into the experiences of people living with the disease. This review summarises the research on the experiences of people living with LF disability. The review highlights the widespread social stigma and oppressive psychological issues that face most people living with LF-related disability. Physical manifestations of LF make daily activities and participation in community life difficult. The findings confirm the need for the Global Programme to Eliminate Lymphatic Filariasis (GPELF) to support morbidity management activities that address the complex biopsychosocial issues that people living with LF-related disability face. 相似文献
54.
Colleen L. Lau Kimberly Y. Won Luke Becker Ricardo J. Soares Magalhaes Saipale Fuimaono Wayne Melrose Patrick J. Lammie Patricia M. Graves 《PLoS neglected tropical diseases》2014,8(11)
Background
As part of the Global Programme to Eliminate Lymphatic Filariasis (LF), American Samoa conducted mass drug administration (MDA) from 2000–2006, and passed transmission assessment surveys in 2011–2012. We examined the seroprevalence and spatial epidemiology of LF post-MDA to inform strategies for ongoing surveillance and to reduce resurgence risk.Methods
ELISA for LF antigen (Og4C3) and antibodies (Wb123, Bm14) were performed on a geo-referenced serum bank of 807 adults collected in 2010. Risk factors assessed for association with sero-positivity included age, sex, years lived in American Samoa, and occupation. Geographic clustering of serological indicators was investigated to identify spatial dependence and household-level clustering.Results
Og4C3 antigen of >128 units (positive) were found in 0.75% (95% CI 0.3–1.6%) of participants, and >32 units (equivocal plus positive) in 3.2% (95% CI 0.6–4.7%). Seroprevalence of Wb123 and Bm14 antibodies were 8.1% (95% CI 6.3–10.2%) and 17.9% (95% CI 15.3–20.7%) respectively. Antigen-positive individuals were identified in all ages, and antibody prevalence higher in older ages. Prevalence was higher in males, and inversely associated with years lived in American Samoa. Spatial distribution of individuals varied significantly with positive and equivocal levels of Og4C3 antigen, but not with antibodies. Using Og4C3 cutoff points of >128 units and >32 units, average cluster sizes were 1,242 m and 1,498 m, and geographical proximity of households explained 85% and 62% of the spatial variation respectively.Conclusions
High-risk populations for LF in American Samoa include adult males and recent migrants. We identified locations and estimated the size of possible residual foci of antigen-positive adults, demonstrating the value of spatial analysis in post-MDA surveillance. Strategies to monitor cluster residents and high-risk groups are needed to reduce resurgence risk. Further research is required to quantify factors contributing to LF transmission at the last stages of elimination to ensure that programme achievements are sustained. 相似文献55.
Cindy Shu Clare Hughes Susan M. Smith Margaret M. Smith Anthony Hayes Bruce Caterson Christopher B. Little James Melrose 《Glycoconjugate journal》2013,30(7):717-725
Composite agarose (1.2 %) polyacrylamide (0.6 %) gel electrophoresis was used to separate discrete populations of native aggrecan and perlecan in newborn to 10 year old ovine intervertebral discs (IVDs). Semi-dry immunoblotting using core-protein and glycosaminoglycan (GAG) side chain specific monoclonal antibodies in combination with chondroitin ABC lyase demonstrated intra-chain native 7-D-4 chondroitin sulphate (CS) sulphation motifs and variable proportions of non-reducing terminal Δ4,5-unsaturated uronate-N-acetylgalactosamine-4-sulphate [2B6(+)] and Δ4,5-unsaturated glucuronate-N-acetylgalactosamine-6-sulphate [3B3(+)] disaccharides. The relative abundance of 2-B-6(+) aggrecan increased with advancing age of the IVD samples while the converse was true for the 3-B-3(+) aggrecan population. Relative 7D4 levels in aggrecan and perlecan were highest in the newborn IVD and significantly lower in the older IVD and other cartilage samples. Quantitation of 7D4 proteoglycan by enzyme linked immunosorbent inhibition assay confirmed the newborn ovine nucleus pulposus (NP) and inner annulus fibrosus (AF) contained higher levels (1.2-1.32 μg 7-D-4-proteoglycan/mg tissue wet weight) than the 2 (0.35-0.42 μg/mg wet weight tissue) and 10 year old IVD samples (0.16-0.22 μg/mg tissue wet weight) with the outer AF zones consistently containing lower levels of 7-D-4 epitope in all cases (P?<?0.001). Cell populations on the margins of the AF and cartilaginous vertebral rudiments in newborn ovine and human foetal IVD strongly expressed 7-D-4 CS epitope and perlecan, This was co-distributed with Notch-1 expression in human foetal IVDs consistent with the 7-D-4 CS sulphation motif representing a marker of tissue development expressed by disc progenitor cell populations. 相似文献
56.
Melrose J Roughley P Knox S Smith S Lord M Whitelock J 《The Journal of biological chemistry》2006,281(48):36905-36914
The aim of this study was to immunolocalize perlecan in human fetal, postnatal, and mature hyaline cartilages and to determine information on the structure and function of chondrocyte perlecan. Perlecan is a prominent component of human fetal (12-14 week) finger, toe, knee, and elbow cartilages; it was localized diffusely in the interterritorial extracellular matrix, densely in the pericellular matrix around chondrocytes, and to small blood vessels in the joint capsules and perichondrium. Aggrecan had a more intense distribution in the marginal regions of the joint rudiments and in para-articular structures. Perlecan also had a strong pericellular localization pattern in postnatal (2-7 month) and mature (55-64 year) femoral cartilages, whereas aggrecan had a prominent extracellular matrix distribution in these tissues. Western blotting identified multiple perlecan core protein species in extracts of the postnatal and mature cartilages, some of which were substituted with heparan sulfate and/or chondroitin sulfate and some were devoid of glycosaminoglycan substitution. Some perlecan core proteins were smaller than intact perlecan, suggesting that proteolytic processing or alternative splicing had occurred. Surface plasmon resonance and quartz crystal microbalance with dissipation experiments demonstrated that chondrocyte perlecan bound fibroblast growth factor (FGF)-1 and -9 less efficiently than endothelial cell perlecan. The latter perlecan supported the proliferation of Baf-32 cells transfected with FGFR3c equally well with FGF-1 and -9, whereas chondrocyte perlecan only supported Baf-32 cell proliferation with FGF-9. The function of perlecan therefore may not be universal but may vary with its cellular origin and presumably its structure. 相似文献
57.
Susan M. Smith John M. Whitelock Renato V. Iozzo Christopher B. Little James Melrose 《Histochemistry and cell biology》2009,132(5):491-503
We evaluated the immunohistochemical distribution of three major proteoglycans of cartilage, i.e., aggrecan, versican and
perlecan vis-a-vis collagens I and II in the developing human spine of first-trimester foetuses. Aggrecan and perlecan were
prominently immunolocalised in the cartilaginous vertebral body rudiments and to a lesser extent within the foetal intervertebral
disc. In contrast, versican was only expressed in the developing intervertebral disc interspace. Using domain-specific monoclonal
antibodies against the various modules of versican, we discovered the V0 isoform as the predominant form present. Versican
immunolocalisations conducted with antibodies directed to epitopes in its N and C termini and GAG-α and GAG-β core protein
domains provided evidence that versican in the nucleus pulposus was either synthesised devoid of a G3 domain or this domain
was proteolytically removed in situ. The V0 versican isoform was localised with prominent fibrillar components in the annular
lamellae of the outer annulus fibrosus. Perlecan was a notable pericellular proteoglycan in the annulus fibrosus and nucleus
pulposus but poorly immunolocalised in the marginal tissues of the developing intervertebral disc, apparently delineating
the intervertebral disc–vertebral body interface region destined to become the cartilaginous endplate in the mature intervertebral
disc. The distribution of collagens I and II in the foetal spine was mutually exclusive with type I present in the outer annulus
fibrosus, marginal tissues around the vertebral body rudiment and throughout the developing intervertebral disc, and type
II prominent in the vertebral rudiment, absent in the outer annulus fibrosus and diffusely distributed in the inner annulus
fibrosus and nucleus pulposus. Collectively, our findings suggest the existence of an intricate and finely balanced interplay
between various proteoglycans and collagens and the spinal cell populations which synthesise and assemble these components
during spinal development. 相似文献
58.
We undertook a comparative immunolocalisation study on type II collagen, aggrecan and perlecan in a number of 12- to 14-week-old
human foetal and postnatal (7–19 months) ovine joints including finger, toe, knee, elbow, hip and shoulder. This demonstrated
that perlecan followed a virtually identical immunolocalisation pattern to that of type II collagen in the foetal tissues,
but a slightly divergent localisation pattern in adult tissues. Aggrecan was also localised in the cartilaginous joint tissues,
which were clearly delineated by toluidine blue staining and the type II collagen immunolocalisations. It was also present
in the capsular joint tissues and in ligaments and tendons in the joint, which stained poorly or not at all with toluidine
blue. In higher power microscopic views, antibodies to perlecan also stained small blood vessels in the synovial lining tissues
of the joint capsule; however, this was not discernable in low power macroscopic views where the immunolocalisation of perlecan
to pericellular regions of cells within the cartilaginous rudiments was a predominant feature. Perlecan was also evident in
small blood vessels in stromal connective tissues associated with the cartilage rudiments and with occasional nerves in the
vicinity of the joint tissues. Perlecan was expressed by rounded cells in the enthesis attachment points of tendons to bone
and in rounded cells in the inner third of the meniscus, which stained prominently with type II collagen and aggrecan identifying
the chondrogenic background of these cells and local compressive loads. Flattened cells within the tendon and in the surface
laminas of articular cartilages and the meniscus did not express perlecan. Collected evidence presented herein, therefore,
indicates that besides being a basement membrane component, perlecan is also a marker of chondrogenic cells in prenatal cartilages.
In postnatal cartilages, perlecan displayed a pericellular localisation pattern rather than the territorial or interterritorial
localisation it displayed in foetal cartilages. This may reflect processing of extracellular perlecan presumably as a consequence
of intrinsic biomechanical loading on these tissues or to divergent functions for perlecan and type II collagen in adult compared
to prenatal tissues. 相似文献
59.
Taylor ML Gutman BL Melrose NA Ingraham AM Schwartz JA Osborn JM 《American journal of botany》2008,95(4):399-413
Cabomba is a small water lily genus that is native to the New World. Studies of pollen development and associated changes in the anther yield valuable characters for considering the evolution of reproductive biology in seed plants. Here we characterized the complete ontogenetic sequence for pollen in Cabomba caroliniana. Anthers at the microspore mother cell, tetrad, free microspore, and mature pollen grain stages were studied using scanning electron, transmission electron, and light microscopy. Tetragonal and decussate tetrads both occur in C. caroliniana, indicating successive microsporogenesis. The exine is tectate-columellate, and the infratectal columellae are the first exine elements to form, followed by a continuous tectum and a thin foot layer. A lamellate endexine initiates in the early free microspore stage, but becomes compressed in mature grains. Tectal microchannels and sculptural rods also initiate during the early free microspore stage, and significant pollenkitt deposition follows, supporting the hypothesis that these elements function in entomophily. The tapetum is morphologically amoeboid, with migratory tapetal cells directly contacting developing free microspores within the anther locule. Results from this study illustrate the importance of including ontogenetic data in analyzing pollen characters and in developing evolutionary and ecological hypotheses. The new palynological data also emphasize the character plasticity that occurs in basal angiosperms. 相似文献
60.
Simon?C?MastbergenEmail author Nathalie?WD?Jansen Johannes?WJ?Bijlsma Floris?PJG?Lafeber 《Arthritis research & therapy》2005,8(1):R2
Treatment of osteoarthritis (OA) with nonsteroidal anti-inflammatory drugs (NSAIDs) diminishes inflammation along with mediators
of cartilage destruction. However, NSAIDs may exert adverse direct effects on cartilage, particularly if treatment is prolonged.
We therefore compared the direct effects of indomethacin, naproxen, aceclofenac and celecoxib on matrix turnover in human
OA cartilage tissue. Human clinically defined OA cartilage from five different donors was exposed for 7 days in culture to
indomethacin, naproxen, aceclofenac and celecoxib – agents chosen based on their cyclo-oxygenase (COX)-2 selectivity. As a
control, SC-560 (a selective COX-1 inhibitor) was used. Changes in cartilage proteoglycan turnover and prostaglandin E2 production were determined. OA cartilage exhibited characteristic proteoglycan turnover. Indomethacin further inhibited proteoglycan
synthesis; no significant effect of indomethacin on proteoglycan release was found, and proteoglycan content tended to decrease.
Naproxen treatment was not associated with changes in any parameter. In contrast, aceclofenac and, prominently, celecoxib
had beneficial effects on OA cartilage. Both were associated with increased proteoglycan synthesis and normalized release.
Importantly, both NSAIDs improved proteoglycan content. Inhibition of prostaglandin E2 production indirectly showed that all NSAIDs inhibited COX, with the more COX-2 specific agents having more pronounced effects.
Selective COX-1 inhibition resulted in adverse effects on all parameters, and prostaglandin E2 production was only mildly inhibited. NSAIDs with low COX-2/COX-1 selectivity exhibit adverse direct effects on OA cartilage,
whereas high COX-2/COX-1 selective NSAIDs did not show such effects and might even have cartilage reparative properties. 相似文献