Bone marrow-derived mesenchymal stem cell(MSC)is one of the most actively studied cell types due to its regenerative potential and immunomodulatory properties.Conventional cell expansion methods using 2D tissue cultur...Bone marrow-derived mesenchymal stem cell(MSC)is one of the most actively studied cell types due to its regenerative potential and immunomodulatory properties.Conventional cell expansion methods using 2D tissue culture plates and 2.5D microcarriers in bioreactors can generate large cell numbers,but they compromise stem cell potency and lack mechanical preconditioning to prepare MSC for physiological loading expected in vivo.To overcome these challenges,in this work,we describe a 3D dynamic hydrogel using magneto-stimulation for direct MSC manufacturing to therapy.With our technology,we found that dynamic mechanical stimulation(DMS)enhanced matrix-integrinβ1 interactions which induced MSCs spreading and proliferation.In addition,DMS could modulate MSC biofunctions including directing MSC differentiation into specific lineages and boosting paracrine activities(e.g.,growth factor secretion)through YAP nuclear localization and FAK-ERK pathway.With our magnetic hydrogel,complex procedures from MSC manufacturing to final clinical use,can be integrated into one single platform,and we believe this‘all-in-one’technology could offer a paradigm shift to existing standards in MSC therapy.展开更多
This study explored methods for improving the performance of Steady-State Visual Evoked Potential(SSVEP)-based Brain-Computer Interfaces(BCI), and introduced a new analytical method to quantitatively analyze and refle...This study explored methods for improving the performance of Steady-State Visual Evoked Potential(SSVEP)-based Brain-Computer Interfaces(BCI), and introduced a new analytical method to quantitatively analyze and reflect the characteristics of SSVEP. We focused on the effect of the pre-stimulation paradigm on the SSVEP dynamic models and the dynamic response process of SSVEP, and performed a comparative analysis of three pre-stimulus paradigms(black, gray, and white). Four dynamic models with different orders(second-and third-order)and with and without a zero point were used to fit the SSVEP envelope. The zero-pole analytical method was adopted to conduct quantitative analysis on the dynamic models, and the response characteristics of SSVEP were represented by zero-pole distribution characteristics. The results of this study indicated that the pre-stimulation paradigm affects the characteristics of SSVEP, and the dynamic models had good fitting abilities with SSVEPs under various types of pre-stimulation. Furthermore, the zero-pole characteristics of the models effectively characterize the damping coefficient, oscillation period, and other SSVEP characteristics. The comparison of zeros and poles indicated that the gray pre-stimulation condition corresponds to a lower damping coefficient, thus showing its potential to improve the performance of SSVEP-BCIs.展开更多
The cellular response to the complex extracellular microenvironment is highly dynamic in time and type of extracellular matrix.Accurately reconstructing this process and analyzing the changes in receptor conformation ...The cellular response to the complex extracellular microenvironment is highly dynamic in time and type of extracellular matrix.Accurately reconstructing this process and analyzing the changes in receptor conformation on the cell membrane surface and intracellular or intercellular signaling has been a major challenge in analytical chemistry and biophysical methodology.In this paper,a time-coded multiconcentration microfluidic chemical waveform generator was developed for the dynamic signaling probing with single-cell array of high temporal resolution,high throughput,and multi-concentration combination stimulation.Based on innovative microchannel structure,sophisticated external control methods and multiplexing technology,the system not only allowed for temporally sequential permutations of the four concentrations of stimuli(time code),but also generated pulsed and continuous waveforms at different frequencies in a highly controllable manner.Furthermore,the single-cell trap array was set up to efficiently capture cells in suspension,dramatically increasing throughput and reducing experiment preparation time.The maximum frequency of the platform was 1 Hz,and one cell could be stimulated at multiple frequencies.To show the ability of the system to investigate rapid biochemical events in high throughput,pulse stimulation and continuous stimulation of different frequencies and different time codes,combined with four concentrations of histamine(HA),were generated for probing G protein-coupled receptor(GPCR)signaling in He La cells.Then,statistical analysis was performed for the mean peak height and mean peak area of the cellular response.We believe that the time-coded multi-concentration microfluidic chemical waveform generator will provide a novel strategy for analytical chemistry,biophysics,cell signaling,and individualized medicine applications.展开更多
The effect of different extraction and purification conditions on the microbial protein obtained from solid-state fermentation(SSF)and the effect of periodical dynamic changes of air on protein have been studied.The m...The effect of different extraction and purification conditions on the microbial protein obtained from solid-state fermentation(SSF)and the effect of periodical dynamic changes of air on protein have been studied.The mechanism of solid-state fermentation with periodical dynamic changes of air is also discussed.Compared with static solid-state fermentation,periodical dynamic changes of air afford a higher protein mass;from 1 g of the fermentation microbe,5.3 mg of the intracellular protein(an increase of 34.63%)and 9.09 mg of the extracellular protein(an increase of 17.8%)were obtained on the sixth day of fermentation.The filter paper activity and carboxymethyl cellulose activity(FPA and CMCase)of the extracellular protein are 1.739μmol/s and 109.592μmol/s,respectively,which represent increases of 60.1%and 21.2%over the corresponding values for static solid-state fermentation.The FPA and CMCase of the intra-cellular protein are 0.245μmol/s and 6.392μmol/s,respec-tively,which represent decreases of 22.2%and 38.7%over the corresponding values for static solid-state fermentation.The enzyme activity of the microbial extracellular protein in solid-state fermentation with periodical dynamic changes of air on the fifth day is nearly equal to that on the sixth day without periodical dynamic changes of air,so the period of fermentation can be shortened.The sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE)experiments suggest that pulsating air pressure stimulation leads to a decrease in the amount of protein component with molecule mass of about 80400Da,and an increase in the amount of protein component with molecule mass of about 28520Da.展开更多
基金supported by NUS Presidential Young Professorship,MOE Tier 1 grantsupported by the NUS Research Scholarship.
文摘Bone marrow-derived mesenchymal stem cell(MSC)is one of the most actively studied cell types due to its regenerative potential and immunomodulatory properties.Conventional cell expansion methods using 2D tissue culture plates and 2.5D microcarriers in bioreactors can generate large cell numbers,but they compromise stem cell potency and lack mechanical preconditioning to prepare MSC for physiological loading expected in vivo.To overcome these challenges,in this work,we describe a 3D dynamic hydrogel using magneto-stimulation for direct MSC manufacturing to therapy.With our technology,we found that dynamic mechanical stimulation(DMS)enhanced matrix-integrinβ1 interactions which induced MSCs spreading and proliferation.In addition,DMS could modulate MSC biofunctions including directing MSC differentiation into specific lineages and boosting paracrine activities(e.g.,growth factor secretion)through YAP nuclear localization and FAK-ERK pathway.With our magnetic hydrogel,complex procedures from MSC manufacturing to final clinical use,can be integrated into one single platform,and we believe this‘all-in-one’technology could offer a paradigm shift to existing standards in MSC therapy.
基金supported by the Key Research and Development Program of Guangdong Province (No. 2018B030339001)the National Key Research and Development Program of China (No. 2017YFB1002505)the National Natural Science Foundation of China (No. 61431007)
文摘This study explored methods for improving the performance of Steady-State Visual Evoked Potential(SSVEP)-based Brain-Computer Interfaces(BCI), and introduced a new analytical method to quantitatively analyze and reflect the characteristics of SSVEP. We focused on the effect of the pre-stimulation paradigm on the SSVEP dynamic models and the dynamic response process of SSVEP, and performed a comparative analysis of three pre-stimulus paradigms(black, gray, and white). Four dynamic models with different orders(second-and third-order)and with and without a zero point were used to fit the SSVEP envelope. The zero-pole analytical method was adopted to conduct quantitative analysis on the dynamic models, and the response characteristics of SSVEP were represented by zero-pole distribution characteristics. The results of this study indicated that the pre-stimulation paradigm affects the characteristics of SSVEP, and the dynamic models had good fitting abilities with SSVEPs under various types of pre-stimulation. Furthermore, the zero-pole characteristics of the models effectively characterize the damping coefficient, oscillation period, and other SSVEP characteristics. The comparison of zeros and poles indicated that the gray pre-stimulation condition corresponds to a lower damping coefficient, thus showing its potential to improve the performance of SSVEP-BCIs.
基金the National Natural Science Foundation of China(Nos.22074047,21775049 and 31700746)the Hubei Provincial Natural Science Foundation of China(No.2020CFB578)the Fundamental Research Funds for Central Universities,HUST(Nos.2020kfy XJJS034 and 2021GCRC056)。
文摘The cellular response to the complex extracellular microenvironment is highly dynamic in time and type of extracellular matrix.Accurately reconstructing this process and analyzing the changes in receptor conformation on the cell membrane surface and intracellular or intercellular signaling has been a major challenge in analytical chemistry and biophysical methodology.In this paper,a time-coded multiconcentration microfluidic chemical waveform generator was developed for the dynamic signaling probing with single-cell array of high temporal resolution,high throughput,and multi-concentration combination stimulation.Based on innovative microchannel structure,sophisticated external control methods and multiplexing technology,the system not only allowed for temporally sequential permutations of the four concentrations of stimuli(time code),but also generated pulsed and continuous waveforms at different frequencies in a highly controllable manner.Furthermore,the single-cell trap array was set up to efficiently capture cells in suspension,dramatically increasing throughput and reducing experiment preparation time.The maximum frequency of the platform was 1 Hz,and one cell could be stimulated at multiple frequencies.To show the ability of the system to investigate rapid biochemical events in high throughput,pulse stimulation and continuous stimulation of different frequencies and different time codes,combined with four concentrations of histamine(HA),were generated for probing G protein-coupled receptor(GPCR)signaling in He La cells.Then,statistical analysis was performed for the mean peak height and mean peak area of the cellular response.We believe that the time-coded multi-concentration microfluidic chemical waveform generator will provide a novel strategy for analytical chemistry,biophysics,cell signaling,and individualized medicine applications.
基金supported by the National Basic Research Program of China(973 Program)(Grant No.2004CB719700)CAS Knowledge Innovation Program Important Direction Project(Grant No.KJCX2-SW·206)Hi-Tech Research and Development Program of China(863 program)(Grant No.2003AA514023).
文摘The effect of different extraction and purification conditions on the microbial protein obtained from solid-state fermentation(SSF)and the effect of periodical dynamic changes of air on protein have been studied.The mechanism of solid-state fermentation with periodical dynamic changes of air is also discussed.Compared with static solid-state fermentation,periodical dynamic changes of air afford a higher protein mass;from 1 g of the fermentation microbe,5.3 mg of the intracellular protein(an increase of 34.63%)and 9.09 mg of the extracellular protein(an increase of 17.8%)were obtained on the sixth day of fermentation.The filter paper activity and carboxymethyl cellulose activity(FPA and CMCase)of the extracellular protein are 1.739μmol/s and 109.592μmol/s,respectively,which represent increases of 60.1%and 21.2%over the corresponding values for static solid-state fermentation.The FPA and CMCase of the intra-cellular protein are 0.245μmol/s and 6.392μmol/s,respec-tively,which represent decreases of 22.2%and 38.7%over the corresponding values for static solid-state fermentation.The enzyme activity of the microbial extracellular protein in solid-state fermentation with periodical dynamic changes of air on the fifth day is nearly equal to that on the sixth day without periodical dynamic changes of air,so the period of fermentation can be shortened.The sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE)experiments suggest that pulsating air pressure stimulation leads to a decrease in the amount of protein component with molecule mass of about 80400Da,and an increase in the amount of protein component with molecule mass of about 28520Da.
