AIM:To study the bacteriocidal or bacteriostatic role of mast cells during infection with Mycobacterium.METHODS:Mycobacterium marinum(M.marinum)(BAA-535/M strain)was investigated for its ability to grow at a temperatu...AIM:To study the bacteriocidal or bacteriostatic role of mast cells during infection with Mycobacterium.METHODS:Mycobacterium marinum(M.marinum)(BAA-535/M strain)was investigated for its ability to grow at a temperature relevant to the mammalian host.Primary mast cells were differentiated from bone marrows of mice,a human mast cell line(HMC-1)and a human monocytic cell line(Mono Mac6)were maintained in culture.Mice were stimulated by intraperitoneal injection of heat-killed M.marinum to study cytochemically the degranulation of peritoneal mast cells.HMC-1 cells were stimulated with M.marinum to analyse m RNA expression for inflammatory reactant genes,while HMC-1 and primary mouse mast cells were infected with M.marinum to establish in parallel cell viability(lactate dehydrogenase release and cell counts)and viable mycobacterial counts.Flow cytometry was used to assess intracellular presence of fluorescein isothiocyanate labelled M.marinum after trypan blue quenching and to measure the extent of infection-induced apoptosis or necrosis in HMC-1.A GFP expressing recombinant M.marinum strain was used to assess intracellular location by fluorescence microscopy.Light microscopy of osmium tetroxide and Gram Twortstained sections of 0.5 μm and transmission electron microscopy were undertaken as sensitive methods.RESULTS:Since its isolation,M.marinum has adapted to grow at 37 ℃.This study found that M.marinum infects HMC-1 cells and primary murine mast cells,where they survive,replicate,and cause dose dependent cell damage over the analysis period of up to 120 h.Amikacin was an effective aminoglycoside antibiotic to eliminate extracellular or membrane attached M.marinum in order to adequately quantify the intracellular bacterial loads.In vivo,intraperitoneal injection of heat-killed M.marinum led to the release of mast cell granules in mice.HMC-1 cells stimulated with M.marinum showed a biphasic pattern of increased mR NA expression for LL-37 and COX-2/TNF-a during 24 h of stimulation.In HMC-1,M.marinum localised to the cytoplasm whereas in primary mast cells,M.marinum were found in vacuoles.CONCLUSION:The effector role of mast cells in infection with M.marinum can be studied in vitro and in vivo.展开更多
Aim Aging is an independent risk factor for heart disease, however the effective intervention has not been found so far. Bone marrow mesenchymal stem cells (BMSCs) have been shown to offer a wide variety of cel- l...Aim Aging is an independent risk factor for heart disease, however the effective intervention has not been found so far. Bone marrow mesenchymal stem cells (BMSCs) have been shown to offer a wide variety of cel- lular functions including the protective effects on damaged hearts. Here we investigated the antiaging properties of BMSCs and the underlying mechanism in a cellular model of cardiomyocyte senescence and a rat model of aging hearts. Methods In vitro study, neonatal rat ventricular cells (NRVCs) and BMSCs were cocultured in the same dish with a semipermeable membrane to separate the two populations. In vivo, the BMSCs were injected into the rat hearts to observe their antiaging effects. The expression of β-galactosidase and aging-related proteins, and the lev- els of oxidative stress were determined in vivo and in vitro. The heart function was measured by the High-Resolution Imaging System. Results Monocultured NRVCs displayed the senescence-associated phenotypes, characterized by an increase in the number of β-galaetosidase-positive cells and decreases in the degradation and disappearance of cellular organelles in a time-dependent manner. The levels of reactive oxygen species and malondialdehyde were el- evated, whereas the activities of antioxidant enzymes superoxide dismutase and glutathione peroxidase were de- creased, along with upregulation of p53, p21cipl/wafl and p16INK4a in the aging eardiomyoeytes. These deleterious alterations were abrogated in aging NRVCs cocultured with BMSCs. Qualitatively, the same senescent phenotypes were consistently observed in aging rat hearts. Notably, BMSC transplantation significantly prevented these detri- mental alterations and improved the impaired cardiac function in the aging rats. Conclusions BMSCs possess strong antiseneseence action on the aging NRVCs and hearts and can improve cardiac function after transplantation in aging rats. The present study, therefore, provides an alternative approach for the treatment of heart failure in the elderly population.展开更多
The purpose of this paper is to present a new general approach to solve ground-state energies of the double-electron systems in a uniform magnetic field, in which the basic element of evolution is the set in the solut...The purpose of this paper is to present a new general approach to solve ground-state energies of the double-electron systems in a uniform magnetic field, in which the basic element of evolution is the set in the solution space, rather than the point. The paper defines the Cell Evolutionary Algorithm, which implements such a view of the evolution mechanism. First, the optimal set in which the optimal solution may be obtained. Then this approach applies the embedded search method to get the optimal solution. We tested this approach on the atomic structure, and the results show that it can improve not only the efficiency but also the accuracy of the calculations as it relates to this specific problem.展开更多
基金Supported by Faculty for the Future Fellowship grant by the Schlumberger Foundation(recipient Siad S)
文摘AIM:To study the bacteriocidal or bacteriostatic role of mast cells during infection with Mycobacterium.METHODS:Mycobacterium marinum(M.marinum)(BAA-535/M strain)was investigated for its ability to grow at a temperature relevant to the mammalian host.Primary mast cells were differentiated from bone marrows of mice,a human mast cell line(HMC-1)and a human monocytic cell line(Mono Mac6)were maintained in culture.Mice were stimulated by intraperitoneal injection of heat-killed M.marinum to study cytochemically the degranulation of peritoneal mast cells.HMC-1 cells were stimulated with M.marinum to analyse m RNA expression for inflammatory reactant genes,while HMC-1 and primary mouse mast cells were infected with M.marinum to establish in parallel cell viability(lactate dehydrogenase release and cell counts)and viable mycobacterial counts.Flow cytometry was used to assess intracellular presence of fluorescein isothiocyanate labelled M.marinum after trypan blue quenching and to measure the extent of infection-induced apoptosis or necrosis in HMC-1.A GFP expressing recombinant M.marinum strain was used to assess intracellular location by fluorescence microscopy.Light microscopy of osmium tetroxide and Gram Twortstained sections of 0.5 μm and transmission electron microscopy were undertaken as sensitive methods.RESULTS:Since its isolation,M.marinum has adapted to grow at 37 ℃.This study found that M.marinum infects HMC-1 cells and primary murine mast cells,where they survive,replicate,and cause dose dependent cell damage over the analysis period of up to 120 h.Amikacin was an effective aminoglycoside antibiotic to eliminate extracellular or membrane attached M.marinum in order to adequately quantify the intracellular bacterial loads.In vivo,intraperitoneal injection of heat-killed M.marinum led to the release of mast cell granules in mice.HMC-1 cells stimulated with M.marinum showed a biphasic pattern of increased mR NA expression for LL-37 and COX-2/TNF-a during 24 h of stimulation.In HMC-1,M.marinum localised to the cytoplasm whereas in primary mast cells,M.marinum were found in vacuoles.CONCLUSION:The effector role of mast cells in infection with M.marinum can be studied in vitro and in vivo.
文摘Aim Aging is an independent risk factor for heart disease, however the effective intervention has not been found so far. Bone marrow mesenchymal stem cells (BMSCs) have been shown to offer a wide variety of cel- lular functions including the protective effects on damaged hearts. Here we investigated the antiaging properties of BMSCs and the underlying mechanism in a cellular model of cardiomyocyte senescence and a rat model of aging hearts. Methods In vitro study, neonatal rat ventricular cells (NRVCs) and BMSCs were cocultured in the same dish with a semipermeable membrane to separate the two populations. In vivo, the BMSCs were injected into the rat hearts to observe their antiaging effects. The expression of β-galactosidase and aging-related proteins, and the lev- els of oxidative stress were determined in vivo and in vitro. The heart function was measured by the High-Resolution Imaging System. Results Monocultured NRVCs displayed the senescence-associated phenotypes, characterized by an increase in the number of β-galaetosidase-positive cells and decreases in the degradation and disappearance of cellular organelles in a time-dependent manner. The levels of reactive oxygen species and malondialdehyde were el- evated, whereas the activities of antioxidant enzymes superoxide dismutase and glutathione peroxidase were de- creased, along with upregulation of p53, p21cipl/wafl and p16INK4a in the aging eardiomyoeytes. These deleterious alterations were abrogated in aging NRVCs cocultured with BMSCs. Qualitatively, the same senescent phenotypes were consistently observed in aging rat hearts. Notably, BMSC transplantation significantly prevented these detri- mental alterations and improved the impaired cardiac function in the aging rats. Conclusions BMSCs possess strong antiseneseence action on the aging NRVCs and hearts and can improve cardiac function after transplantation in aging rats. The present study, therefore, provides an alternative approach for the treatment of heart failure in the elderly population.
基金Supported by the opening Foundation of state key Laboratory of Magnetic Resonance and Atomic and Molecularphysics, Wuhan Insti
文摘The purpose of this paper is to present a new general approach to solve ground-state energies of the double-electron systems in a uniform magnetic field, in which the basic element of evolution is the set in the solution space, rather than the point. The paper defines the Cell Evolutionary Algorithm, which implements such a view of the evolution mechanism. First, the optimal set in which the optimal solution may be obtained. Then this approach applies the embedded search method to get the optimal solution. We tested this approach on the atomic structure, and the results show that it can improve not only the efficiency but also the accuracy of the calculations as it relates to this specific problem.