Detecting and quantifying intracellular microRNAs(miRNAs)are a critical step in resolving a cancer diagnostic and resolving the ensemble of gene products that orchestrate the living state of cells.However,the nanoprob...Detecting and quantifying intracellular microRNAs(miRNAs)are a critical step in resolving a cancer diagnostic and resolving the ensemble of gene products that orchestrate the living state of cells.However,the nanoprobe for detecting low abundance miRNAs in cell cytosol is restricted by either the"one-to-one"signaltrigger model or di culty for cytosol delivery.To address these challenges,we designed a lightharvesting nanoantenna-based nanoprobe,which directs excitation energy to a single molecule to sensitively detect cytosolic miRNA.With light irradiation,the light-harvesting nanoantenna e ectively disrupted lysosomal structures by generationof reactive oxygen species,substantially achieved cytosol delivery.The nanoantenna containing>4000 donor dyes can e ciently transfer excitation energy to one or two acceptors with 99%e ciency,leading to unprecedented signal amplification and biosensing sensitivity.The designed nanoantenna can quantify cytosolic miR-210 at zeptomolar level.The fluorescence lifetime of the donor exhibited good relationship with miR-210 concentration in the range of 0.032 to 2.97 amol/ngRNA.The zeptomole sensitivity of nanoantenna provides accurate bioimaging of miR-210 both in multiple cell lines and in vivo assay,which creates a pathway for the creation of miRNA toolbox for quantitative epigenetics and personalized medicine.展开更多
Background There were only 3 multiple myeloma (MM) cell lines established in China. In this study,we succeeded in establishing a novel MM cell line and analyzed its biological characteristics. Methods Mononuclear...Background There were only 3 multiple myeloma (MM) cell lines established in China. In this study,we succeeded in establishing a novel MM cell line and analyzed its biological characteristics. Methods Mononuclear cells isolated from the peripheral blood (PB) and bone marrow (BM) of a patient with advanced MM (λ light chain type) were cultured in medium. Cell morphology was analyzed by Wright-Giemsa-staining and cytochemical staining,immunophenotyping by flow cytometry and cytogenetic analysis by chromosome RHG-banding technique. Quantitative fluorescent polymerase chain reaction (PCR) was used to detect Epstein - Barr virus (EBV) DNA. Results The established cell line could survive and proliferate in the presence of feeder cells or conditioned medium. The cells secreted λ light chain and were negative for EBV. The Wright-Giemsa-staining showed typical plasmablast or plasma cell morphology. The cytochemical staining of the cells showed the following reactivity patterns: positive for acid phosphatase,negative for myeloperoxidase. The immunoprofile of the cells was concordant with that of MM cells: positive for CD_ 10 ,CD_ 28 ,CD_ 38 ,CD_ 138 ,CD_ 56 ,CD_ 49d ,CD_ 44 ,CD_ 54 and CD_ 58 ,negative for CD_ 19 , CD_ 40 ,CD_ 95 ,CD_ 95L ,CD_ 34 ,CD_2 and CD_5. The cytogenetic analysis showed complex chromosome abnormality of i (1q+),8q+,13q+,i (17q),i (18q) and +M. There was no difference in morphology,immunophenotype and cytogenetics between cells from PB and BM. Conclusions An MM cell line secreting λ light chain named CZ-1 was established. The cells from both PB and BM have the same biological characteristics.展开更多
In this review, we focused on a few obstacles that hinder three-dimensional （3D） bioprinting process in tissue engineering. One of the obstacles is the bioinks used to deliver cells. Hydrogels are the most widely us...In this review, we focused on a few obstacles that hinder three-dimensional （3D） bioprinting process in tissue engineering. One of the obstacles is the bioinks used to deliver cells. Hydrogels are the most widely used bioink materials; however, they are mechanically weak in nature and cannot meet the requirements for supporting structures, especially when the tissues, such as cartilage, require extracellular matrix to be mechanically strong. Secondly and more importantly, tissue regeneration is not only about building all the components in a way that mimics the structures of living tissues, but also about how to make the constructs function normally in the long term. One of the key issues is sufficient nutrient and oxygen supply to the engineered living constructs. The other is to coordinate the interplays between cells, bioactive agents and extracellular matrix in a natural way. This article reviews the approaches to improve the mechanical strength of hydrogels and their suitability for 3D bioprinting; moreover, the key issues of multiple cell lines coprinting with multiple growth factors, vascularization within engineered living constructs etc. were also reviewed.展开更多
基金supported by start-up fund of Washington State University。
文摘Detecting and quantifying intracellular microRNAs(miRNAs)are a critical step in resolving a cancer diagnostic and resolving the ensemble of gene products that orchestrate the living state of cells.However,the nanoprobe for detecting low abundance miRNAs in cell cytosol is restricted by either the"one-to-one"signaltrigger model or di culty for cytosol delivery.To address these challenges,we designed a lightharvesting nanoantenna-based nanoprobe,which directs excitation energy to a single molecule to sensitively detect cytosolic miRNA.With light irradiation,the light-harvesting nanoantenna e ectively disrupted lysosomal structures by generationof reactive oxygen species,substantially achieved cytosol delivery.The nanoantenna containing>4000 donor dyes can e ciently transfer excitation energy to one or two acceptors with 99%e ciency,leading to unprecedented signal amplification and biosensing sensitivity.The designed nanoantenna can quantify cytosolic miR-210 at zeptomolar level.The fluorescence lifetime of the donor exhibited good relationship with miR-210 concentration in the range of 0.032 to 2.97 amol/ngRNA.The zeptomole sensitivity of nanoantenna provides accurate bioimaging of miR-210 both in multiple cell lines and in vivo assay,which creates a pathway for the creation of miRNA toolbox for quantitative epigenetics and personalized medicine.
文摘Background There were only 3 multiple myeloma (MM) cell lines established in China. In this study,we succeeded in establishing a novel MM cell line and analyzed its biological characteristics. Methods Mononuclear cells isolated from the peripheral blood (PB) and bone marrow (BM) of a patient with advanced MM (λ light chain type) were cultured in medium. Cell morphology was analyzed by Wright-Giemsa-staining and cytochemical staining,immunophenotyping by flow cytometry and cytogenetic analysis by chromosome RHG-banding technique. Quantitative fluorescent polymerase chain reaction (PCR) was used to detect Epstein - Barr virus (EBV) DNA. Results The established cell line could survive and proliferate in the presence of feeder cells or conditioned medium. The cells secreted λ light chain and were negative for EBV. The Wright-Giemsa-staining showed typical plasmablast or plasma cell morphology. The cytochemical staining of the cells showed the following reactivity patterns: positive for acid phosphatase,negative for myeloperoxidase. The immunoprofile of the cells was concordant with that of MM cells: positive for CD_ 10 ,CD_ 28 ,CD_ 38 ,CD_ 138 ,CD_ 56 ,CD_ 49d ,CD_ 44 ,CD_ 54 and CD_ 58 ,negative for CD_ 19 , CD_ 40 ,CD_ 95 ,CD_ 95L ,CD_ 34 ,CD_2 and CD_5. The cytogenetic analysis showed complex chromosome abnormality of i (1q+),8q+,13q+,i (17q),i (18q) and +M. There was no difference in morphology,immunophenotype and cytogenetics between cells from PB and BM. Conclusions An MM cell line secreting λ light chain named CZ-1 was established. The cells from both PB and BM have the same biological characteristics.
文摘In this review, we focused on a few obstacles that hinder three-dimensional （3D） bioprinting process in tissue engineering. One of the obstacles is the bioinks used to deliver cells. Hydrogels are the most widely used bioink materials; however, they are mechanically weak in nature and cannot meet the requirements for supporting structures, especially when the tissues, such as cartilage, require extracellular matrix to be mechanically strong. Secondly and more importantly, tissue regeneration is not only about building all the components in a way that mimics the structures of living tissues, but also about how to make the constructs function normally in the long term. One of the key issues is sufficient nutrient and oxygen supply to the engineered living constructs. The other is to coordinate the interplays between cells, bioactive agents and extracellular matrix in a natural way. This article reviews the approaches to improve the mechanical strength of hydrogels and their suitability for 3D bioprinting; moreover, the key issues of multiple cell lines coprinting with multiple growth factors, vascularization within engineered living constructs etc. were also reviewed.
