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1.
2.

Introduction

Guided treatments with nanoparticles and cold atmospheric plasma are a new approach in cancer therapy. Plasma is an ionized gas that has reactive and energetic particles and can be produced in the laboratory by different methods.

Material and methods

Plasma jet therapy was employed to irradiate HCT-116 cells (human colorectal cancer cells) which were cultured in the presence of gold nanoparticles (GNPs). Cell cytotoxicity was tested with 3-[4, 5-dimethylthiazol-2-yl]-2,5 diphenyltetrazolium bromide (MTT), and cancerous cell apoptosis was shown by 4’,6-diamidino-2-phenylindole (DAPI) staining.

Results

The results showed that cell death was increased significantly with p < 0.001 by cold atmospheric plasma in the presence of gold nanoparticles.

Conclusions

It appears that non-thermal plasma and gold nanoparticles synergism is a promising approach in colon cancer therapy.  相似文献   

3.
Macrophages can be obtained about 99 per cent pure and in a yield of about 50 per cent by ficoll (polysucrose) density gradient centrifugation of mouse peritoneal exudate cells. The macrophages appear in the lighter fractions. The lymphocytes appear in the denser fractions and can be further purified on a polystyrene bead, rayon wool column. The method also separates guinea-pig and rat peritoneal exudate cells.

Peritoneal exudate macrophages take up colloidal radioactive gold in vivo. However, macrophages differ in the ability to take up colloidal gold and the macrophages which phagocytose the most gold appear in the lightest fraction after density gradient centrifugation.

  相似文献   

4.
The pinocytosis of radioactive colloidal gold by peritoneal exudate cells from guinea-pigs sensitized against tuberculin or ovalbumin was enhanced by specific antigen. The effect was mediated by soluble material (gold uptake stimulator (GUS) produced by lymphocytes and acting upon macrophages. GUS production began within the first 18 hours of culture and required 24 hours contact to maximally stimulate normal peritoneal exudate cells.

Comparison of the gold uptake and migration inhibition assays on identical populations of sensitive exudate cells showed in terms of minimum effective antigen dose that the gold uptake assay was considerably more sensitive. Similarly, when sensitive lymph node cells were incubated with antigen, activity could be detected in the culture supernatants at about sixty-four-fold greater dilution by the uptake of gold.

At low concentrations of GUS a log-linear dose response relationship held. At high concentrations, gold uptake levelled off or even fell, and this was true for concentrations of antigen greater than optimal when added to sensitive exudate cells.

Chromium uptake varied little at the concentrations of antigen and supernatant factors tested. Hence the effects of gold uptake are unlikely to be indirect effects on cell proliferation or viability.

  相似文献   

5.

Objective

We have previously demonstrated the efficient and time-dependent transvascular localization of Sialyl Lewis X (SLX)-liposomes to inflammatory sites, but the final target of the SLX-liposomes remained uncertain. The aim of this study was to identify the target cells of the liposomes within the inflamed joints of collagen antibody-induced arthritis (CAIA) model mice.

Methods

SLX-liposomes and unlabeled liposomes encapsulating high-density colloidal gold were administered intravenously into the caudal vein of CAIA mice on day 5 after induction of arthritis when the inflammatory score was maximal (n = 6 per group). Six hours or 24 h after liposome administration, animals were euthanized and hind limbs and ankles were excised without perfusion. After fixation, synovial tissues were examined by light microscopy after silver enhancement of colloidal gold or by transmission electron microscopy.

Results

Silver-enhanced signals were detected within the cells around E-selectin-positive blood vessels in the synovium of the SLX-liposome group. These cells were positive for the macrophage/monocyte marker F4/80 or neutrophil marker Ly-6G. Transmission electron microscopy detected the colloidal gold signals together with liposome-like structures within the phagosomes of synovial macrophages. Transmission electron microscopy and energy dispersive X-ray spectrometry could determine gold elements in the lysosomes of synovial macrophages.

Conclusions

The results of the current study demonstrate that SLX-liposomes primarily targeting E-selectin in activated endothelial cells could potentially deliver their contents into inflammatory cells around synovial blood vessels in arthritic joints.  相似文献   

6.
Mosquito-borne diseases in India, e.g., malaria, dengue, chikungunya, filariasis, and Japanese encephalitis cause thousands of deaths per year. Mosquito control is to enhance the health and quality of life of county residents and visitors through the reduction of mosquito populations. Mosquito control is of serious concern in developing countries like India due to the lack of general awareness, development of resistance, and socioeconomic reasons. Noble metal nanoparticles have been used because of their unique optical properties; especially gold and silver have a broad absorption band in the visible region of electromagnetic spectrum. Synthesis of gold nanoparticles using Cymbopogan citratus is an ecofriendly approach for safer environment. C. citratus leaf broth was a good reducing agent that converted chloroauric acid (HAuCl4) to metal gold and further heating converted it into nanoparticles. Characterization using UV spectrophotometer, X-ray diffraction, Fourier transform infrared spectroscopy, particle size analyzer, and transmission electron microscopy confirmed that the particles are gold nanoparticles ranging between 10 and 110 nm with an average particles size of 20 nm. Further biosynthesized gold nanoparticles and Anthocephalus cadamba were experimented for the larvicidal effect on the filarial vector, Culex quinquefasciatus. Results showed that the gold nanoparticles are much toxic than the plant extract. Observed lethal concentrations (LC50 and LC90) were 1.08 and 2.76 ppm for gold nanoparticles and 21.82 and 79.52 ppm for the third instar of C. quinquefasciatus.  相似文献   

7.
Sitter  H.  Torossian  A.  Duda  D.  Sattler  J. 《Inflammation research》2004,53(2):S164-S168
Objective:Histamine release may cause anaphylactoid reactions. However, during anaesthesia and surgery especially cardiovascular effects may not be regarded as histamine-related. Therefore, we adapted the classical concept of histamine release reactions to the perioperative situation and validated the new paradigm. Methods:Elevated plasma histamine (diagnostic gold standard) was correlated to potentially related intraoperative signs and symptoms. The validity, repeatability and sensitivity of the ‘gold standard’ was tested by ROC analysis in volunteers, who also received H1-/H2-histamine antagonists. Additionally, a dose-response relationship was determined in dogs using the histamine releaser compound 48/80. Results:The ‘gold standard’ had a sensitivity of 96% (90%–100%) and a specificity of 93% (85%–100%). The reproducibility was proven by repeated injections of histamine. Skin reactions, tachycardia and hypertension were identified as histamine-related diagnostic variables. A dose-response curve of plasma histamine release was created. Conclusions:The defined ‘gold standard’ is valid for the diagnosis of histamine-related reactions during anaesthesia and surgery. It may help to identify patients, who could benefit from pre-anaesthetic antihistamine prophylaxis.  相似文献   

8.
H Yeger  V I Kalnins 《Virology》1978,91(2):489-492
An indirect immunocytochemical labeling technique employing antibody bound to colloidal gold particles as a marker was used to study the distribution of the major viral glycoprotein gp70 on surfaces of cells and virus in cultures infected with wild-type and mutant ts29 Rauscher murine leukemia virus. The temperature-sensitive mutant-infected cells grown at the nonpermissive temperature acquire submembranous densities which are believed to contain the gag gene-coded uncleaved polypeptide precursor. The cell membranes associated with these densities are heavily labeled with the gold-colloid marker and possess much larger quantities of gp70 than the rest of the cell surface. The budding and extracellular virions produced by these cells after shiftdown to the permissive temperature are also heavily labeled. These findings are consistent with the view that structural protein-membrane-glycoprotein interactions are important in type C virus assembly.  相似文献   

9.
Airway epithelial cells act as the first barrier against pathogens. These cells recognize conserved structural motifs expressed by microbial pathogens via Toll-like receptors (TLRs) expressed on the surface. In contrast to the level of expression in lymphoid cells, the level of expression of TLR2 and TLR4 in airway epithelial cells is low under physiological conditions. Here we explored whether Klebsiella pneumoniae upregulates the expression of TLRs in human airway epithelial cells. We found that the expression of TLR2 and TLR4 by A549 cells and human primary airway cells was upregulated upon infection with K. pneumoniae. The increased expression of TLRs resulted in enhancement of the cellular response upon stimulation with Pam3CSK4 and lipopolysaccharide, which are TLR2 and TLR4 agonists, respectively. Klebsiella-dependent upregulation of TLR expression occurred via a positive IκBα-dependent NF-κΒ pathway and via negative p38 and p44/42 mitogen-activated protein kinase-dependent pathways. We showed that Klebsiella-induced TLR2 and TLR4 upregulation was dependent on TLR activation. An isogenic capsule polysaccharide (CPS) mutant did not increase TLR2 and TLR4 expression. Purified CPS upregulated TLR2 and TLR4 expression, and polymyxin B did not abrogate CPS-induced TLR upregulation. Although no proteins were detected in the CPS preparation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and colloidal gold staining, we could not rule out the possibility that traces of protein in our CPS preparation could have been responsible, at least in part, for the TLR upregulation.  相似文献   

10.
We prepared pH-sensitive drug–dendrimer conjugate-hybridized gold nanorod as a promising platform for combined cancer photothermal-chemotherapy under in vitro and in vivo conditions. Poly(ethylene glycol)-attached PAMAM G4 dendrimers (PEG–PAMAM) were first covalently linked on the surface of mercaptohexadecanoic acid-functionalized gold nanorod (MHA-AuNR), with subsequent conjugation of anti-cancer drug doxorubicin (DOX) to dendrimer layer using an acid-labile-hydrazone linkage to afford PEG–DOX–PAMAM–AuNR particles. The particles with a high PEG–PAMAM dendrimer coverage density (0.28 per nm2 AuNR) showed uniform sizes and excellent colloidal stability. In vitro drug release studies demonstrated that DOX released from PEG–DOX–PAMAM–AuNR was negligible under normal physiological pH, but it was enhanced significantly at a weak acidic pH value. The efficient intracellular acid-triggered DOX release inside of lysosomes was confirmed using confocal laser scanning microscopy analysis. Furthermore, the combined photothermal-chemo treatment of cancer cells using PEG–DOX–PAMAM–AuNR for synergistic hyperthermia ablation and chemotherapy was demonstrated both in vitro and in vivo to exhibit higher therapeutic efficacy than either single treatment alone, underscoring the great potential of PEG–DOX–PAMAM–AuNR particles for cancer therapy.  相似文献   

11.
Phospholipase C (PLC)β has a role in saliva secretion by controlling intracellular Ca2+ via its product, IP3. The present study was attempted to localize PLCβ isoforms in mouse salivary glands in situ. A single major band was detected for PLCβ3 in immunoblots of the parotid and sublingual glands (PG, SLG), while no such band was seen in the submandibular gland (SMG). No bands were detected for PLCβ1 or 4 in the three glands. In immuno-light microscopy of PG and SLG, substantial immunoreactivity for PLCβ3 was seen in the cytoplasm including the plasmalemma of almost all ductal cells, while no distinct immunoreactivity was discerned in most acinar cells except for sublingual demilune cells. Numerous ductal cells exhibited higher immunoreactivity for PLCβ3 in their apical/supranuclear cell domain including the plasmalemma than in the basal/infranuclear domain, indicating an apico-basal polarity. In immuno-gold electron microscopy of PG ducts and SLG ducts and demilunes, most gold particles were found in association with plasma membranes as well as various intracellular membranes, most of which formed small oblong or flattened vesicles and vacuoles. A few particles were seen without association with any membranous structures. The present finding supports the previous physio-pharmacological result that Ca2+-signaling proteins as well as initial intracellular Ca2+ changes occur in the apical cell domain including the plasma membranes of the exocrine cells.  相似文献   

12.
Lysosomal imaging represents a potent tool for investigating the organization of related cellular events and their modulation via diagnostic and therapeutic approaches. However, specific labeling of the lysosome in live cells is a significant challenge. Taking advantage of the inherent lysosomal entry of nanoparticles and unique digestive inclusions in the lysosome, we developed a nanoparticle-based, enzyme-switchable fluorescence OFF-ON strategy for specific labeling of the lysosome and further imaging of extracellular acidification-induced lysosome trafficking in living cells. The nanoprobe comprised a 16 nm spherical gold nanoparticle as the core and an enzyme-responsive oligomer of fluorescein-conjugated oligo(4-vinyl-phenyl phosphate) as the shell. Due to quenching of the core gold nanoparticle, the nanoprobe was non-fluorescent. After incubation with cancer cells, the nanoprobe was rapidly internalized via scavenger receptor-mediated endocytosis and significantly shuffled into the lysosome. The nanoprobe specifically lighted up the lysosome owing to lysosome-induced fluorescence enhancement. Specifically, digestive inclusions in the lysosome hydrolyzed and released gold-quenched fluorescein molecules, leading to significant augmentation of fluorescence. On account of specific lysosomal labeling, the nanoprobe effectively facilitated imaging of a 4–6 μm anterograde trafficking event of the lysosome from the perinuclear region to the cell surface when an acidic extracellular environment developed. Our findings collectively highlight the use of nanoprobes for lysosomal imaging.  相似文献   

13.
Background: There is no single technique that can meet the criteria in identification of Helicobacter pylori. The diagnosis is important asantimicrobial resistance is frequently observed and associated with treatment failure. The present study was conducted to evaluate diagnostic tests for identification of H pylori and to assess their antimicrobial resistance pattern. Materials and Methods: Biopsies of gastric tissue from 200 patients with disorders of the upper gastrointestinal tract were studied for detection of H pylori by various methods like culture, H and E staining and urease test. Antimicrobial susceptibility testing was carried out by Kirby Bauer’s disc diffusion method. Results: Out of 200 patients, H pylori was detected by rapid urease test, H and E staining and culture in 26.5%, 14.5% and 2.5% cases respectively. H and E was taken as the gold standard. Sensitivity of urease test was 76.6% and of culture 13.3%. Specificity of urease was 81.7% in comparison with culture which showed 99.4% specificity. Metronidazole (05) showed high level of resistance followed by amoxicillin (03) and norfloxacillin (03). Tetracycline, erythromycin, levofloxacin and cotrimoxazole showed one resistance each to H pylori. Conclusion: H and E is taken as the gold standard according to CDC. Urease test is a better screening procedure than culture. H pylori resistance to metronidazole in our zone was highest. This is due to general and extensive use of metronidazole for other infectious diseases. Our study suggests need for a systematic approach to determine antibiogram of the strains before considering the drug regimens.  相似文献   

14.

Background

Electronic patient records (EPRs) contain a large amount of information written in free text. This information is considered very valuable for research but is also very sensitive since the free text parts may contain information that could reveal the identity of a patient. Therefore, methods for de-identifying EPRs are needed. The work presented here aims to perform a manual and automatic Protected Health Information (PHI)-annotation trial for EPRs written in Swedish.

Methods

This study consists of two main parts: the initial creation of a manually PHI-annotated gold standard, and the porting and evaluation of an existing de-identification software written for American English to Swedish in a preliminary automatic de-identification trial. Results are measured with precision, recall and F-measure.

Results

This study reports fairly high Inter-Annotator Agreement (IAA) results on the manually created gold standard, especially for specific tags such as names. The average IAA over all tags was 0.65 F-measure (0.84 F-measure highest pairwise agreement). For name tags the average IAA was 0.80 F-measure (0.91 F-measure highest pairwise agreement). Porting a de-identification software written for American English to Swedish directly was unfortunately non-trivial, yielding poor results.

Conclusion

Developing gold standard sets as well as automatic systems for de-identification tasks in Swedish is feasible. However, discussions and definitions on identifiable information is needed, as well as further developments both on the tag sets and the annotation guidelines, in order to get a reliable gold standard. A completely new de-identification software needs to be developed.  相似文献   

15.
A study was conducted to determine the ability of the inclusion immunofluorescence assay (inclusion IFA) to act as a screening test to detect samples with antibodies to Chlamydia pneumoniae; microimmunofluorescence (MIF) was used as the “gold standard.” In addition, the inclusion IFA was compared using HEp-2 cells infected with either C. pneumoniae CM-1 or Chlamydia trachomatis serovar E. A total of 331 serum samples representing a range of MIF titers were evaluated. The sensitivities of the inclusion IFA for detecting samples with C. pneumoniae MIF titers of ≥16 were 96.9 and 74.8% with C. pneumoniae- and C. trachomatis-infected cells, respectively. For samples with an elevated C. pneumoniae MIF titer of ≥512, the sensitivities of the C. pneumoniae- and C. trachomatis-based inclusion IFA were 97.0 and 8.8%, respectively. These results suggest that the inclusion IFA is not a genus-specific test, as evidenced by the failure of the C. trachomatis-infected cells to detect a significant number of samples with C. pneumoniae antibodies. Samples that had elevated C. pneumoniae inclusion IFA and MIF titers but that were found negative (titer, <16) by the C. trachomatis inclusion IFA were further tested by an in vitro neutralization assay for functional antibodies that might not have been detected by the serological assays. The in vitro neutralization results corroborated the serological results in that all seven sera tested had a neutralization titer for C. pneumoniae (range, 20 to 225), while all but one failed to have any effect on the infectivity of C. trachomatis serovar E. While the C. pneumoniae inclusion IFA had a high sensitivity for detecting chlamydial antibodies, depending on whether it was used as a screening test for detecting samples with low (≥16) or elevated (≥512) MIF titers, its specificity ranged from 53.4 to 77.1%. In conclusion, the inclusion IFA with C. pneumoniae-infected cells was best suited as a sensitive screening test for identifying specimens with elevated MIF titers (those associated with a possible acute infection with C. pneumoniae).  相似文献   

16.
The design and development of functional hybrid nanomaterials is currently a topic of great interest in biomedicine. Herein we investigated the grafting of Ru(II) polypyridyl complexes onto gold nanospheres (Ru@AuNPs) to improve the particles' near infrared (NIR) absorption, and ultimately allow for application in photothermal cancer therapy. As demonstrated in this article, these ruthenium(II) complexes could indeed significantly enhance gold nanospheres' two-photon luminescence (PTL) intensity and photothermal therapy (PTT) efficiency. The best dual functional nanoparticles of this study were successfully used for real-time luminescent imaging-guided PTT in live cancer cells. Furthermore, in vivo tumor ablation was achieved with excellent treatment efficacy under a diode laser (808 nm) irradiation at the power density of 0.8 W/cm2 for 5 min. This study demonstrates that the coupling of inert Ru(II) polypyridyl complexes to gold nanospheres allows for the enhancement of two-photon luminescence and for efficient photothermal effect.  相似文献   

17.
An unusual case of bone marrow dysplasia is reported. The features of particular interest are the very high incidence of nuclear drumsticks on the polymorphs, a curious appearance of these same nuclei on electron microscopy, and C trisomy of the bone marrow cells; it is possible that the condition resulted from gold and/or x-ray treatment.  相似文献   

18.
The identification of Campylobacter species and related organisms at the species level has always been difficult using phenotypic methods because of their low metabolic activity, whereas molecular methods are more reliable but time-consuming. In this study, 1007 different strains were identified using three different methods: conventional methods, molecular biology (real-time PCR and sequencing) and matrix assisted laser desorption ionization–time of flight (MALDI-TOF) mass spectrometry. Molecular methods were considered the gold standard. The accuracy of MALDI-TOF mass spectrometry reached 100% compared with the gold standard for all of the Campylobacter species, except Campylobacter jejuni (99.4%). The accuracy of conventional methods compared with the gold standard ranged from 0% to 100% depending on the species. However, MALDI-TOF mass spectrometry was not able to identify a mixture of two different species present in the same sample in four instances. Finally, MALDI-TOF mass spectrometry is highly recommended to identify Campylobacter spp. as only 0.4% discrepancy was found, whereas conventional methods led to 4.5% discrepancy.  相似文献   

19.
The present report describes the synthesis and biological evaluation of a molecular imaging platform based on gold nanoparticles directly labeled with indium-111. The direct labeling approach facilitated radiolabeling with high activities while maintaining excellent stability within the biological environment. The resulting imaging platform exhibited low interference of the radiolabel with targeting molecules, which is highly desirable for in-vivo probe tracking and molecular targeted tumor imaging. The indium-111 labeled gold nanoparticles were synthesized using a simple procedure that allowed stable labeling of the nanoparticle core with various indium-111 activities. Subsequent surface modification of the particle cores with RGD-based ligands at various densities allowed for molecular targeting of the αvß3 integrin in-vitro and for molecular targeted imaging in human melanoma and glioblastoma models in-vivo. The results demonstrate the vast potential of direct labeling with radioisotopes for tracking gold nanoparticles within biological systems.  相似文献   

20.
Haemophilus influenzae is a significant causative agent of respiratory tract infections (RTI) worldwide. The development of a rapid H. influenzae diagnostic assay that would allow for the implementation of infection control measures and also improve antimicrobial stewardship for patients is required. A number of nucleic acid diagnostics approaches that detect H. influenzae in RTIs have been described in the literature; however, there are reported specificity and sensitivity limitations for these assays. In this study, a novel real-time PCR diagnostic assay targeting the smpB gene was designed to detect all serogroups of H. influenzae. The assay was validated using a panel of well-characterized Haemophilus spp. Subsequently, 44 Haemophilus clinical isolates were collected, and 36 isolates were identified as H. influenzae using a gold standard methodology that combined the results of matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) and a fucK diagnostic assay. Using the novel smpB diagnostic assay, 100% concordance was observed with the gold standard, demonstrating a sensitivity of 100% (95% confidence interval [CI], 90.26% to 100.00%) and a specificity of 100% (95% CI, 63.06% to 100.00%) when used on clinical isolates. To demonstrate the clinical utility of the diagnostic assay presented, a panel of lower RTI samples (n = 98) were blindly tested with the gold standard and smpB diagnostic assays. The results generated were concordant for 94/98 samples tested, demonstrating a sensitivity of 90.91% (95% CI, 78.33% to 97.47%) and a specificity of 100% (95% CI, 93.40% to 100.00%) for the novel smpB assay when used directly on respiratory specimens.  相似文献   

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