Human pluripotent stem cells(hPSCs), including human embryonic stem cells and human induced pluripotent stem cells, are promising sources for hematopoietic cells due to their unlimited growth capacity and the pluripot...Human pluripotent stem cells(hPSCs), including human embryonic stem cells and human induced pluripotent stem cells, are promising sources for hematopoietic cells due to their unlimited growth capacity and the pluripotency. Dendritic cells(DCs), the unique immune cells in the hematopoietic system, can be loaded with tumor specific antigen and used as vaccine for cancer immunotherapy. While autologous DCs from peripheral blood are limited in cell number, hPSC-derived DCs provide a novel alternative cell source which has the potential for large scale production. This review summarizes recent advances in differentiating hPSCs to DCs through the intermediate stage of hematopoietic stem cells. Step-wise growth factor induction has been used to derive DCs from hPSCs either in suspension cultureof embryoid bodies(EBs) or in co-culture with stromal cells. To fulfill the clinical potential of the DCs derived from hPSCs, the bioprocess needs to be scaled up to produce a large number of cells economically under tight quality control. This requires the development of novel bioreactor systems combining guided EB-based differentiation with engineered culture environment. Hence, recent progress in using bioreactors for hPSC lineage-specific differentiation is reviewed. In particular, the potential scale up strategies for the multistage DC differentiation and the effect of shear stress on hPSC differentiation in bioreactors are discussed in detail.展开更多
The influence of cells groupings factor to the performance of the cells groupings time-shift pilot scheme is researched for the multiple cells large scale antennas systems（LSAS）. The former researches have confirmed...The influence of cells groupings factor to the performance of the cells groupings time-shift pilot scheme is researched for the multiple cells large scale antennas systems（LSAS）. The former researches have confirmed that the cells groupings time-shift pilots scheme is effective to reduce inter-cell interference, especially pilot contamination, which results from the pilot reuse in adjacent cells. However, they have not specified reasonable cells groupings factor, which plays a critical role in the general performance of the LSAS. Therefore, this problem is researched in details. The time for reverse-link data transmission will be compressed, when the groupings factor surpasses a certain range. Thus it is not always beneficial to increase the cells groupings factor without limitation. Furthermore,a reasonable cells groupings factor is deduced from the perspective of optimization to enhance the system performance. Simulations verify the proposed cell grouping factor.展开更多
The catalyst layer （CL） of proton exchange mem-brane fuel cell （PEMFC） involves various particles and pores in meso-scale, which has an important effect on the mass, charge （proton and electron） and heat transpo...The catalyst layer （CL） of proton exchange mem-brane fuel cell （PEMFC） involves various particles and pores in meso-scale, which has an important effect on the mass, charge （proton and electron） and heat transport coupled with the electrochemical reactions. The coarse-grained molecular dynamics （CG-MD） method is employed as a meso-scale structure reconstruction technique to mimic the self-organization phenomena in the fabrication steps of a CL. The meso-scale structure obtained at the equilibrium state is further analyzed by molecular dynamic （MD） software to provide the necessary microscopic parameters for understanding of multi-scale and-physics processes in CLs. The primary pore size distribution （PSD） and active platinum （Pt） surface areas are also calculated and then compared with the experiments. In addition, we also highlight the implementation method to combine microscopic elementary kinetic reaction schemes with the CG-MD approaches to provide insight into the reactions in CLs. The concepts from CG modeling with particle hydrodynamics （SPH） and the problems on coupling of SPH with finite element modeling （FEM） methods are further outlined and discussed to understand the effects of the meso-scale transport phenomena in fuel cells.展开更多
We describe the microfabrication of ^85Rb vapour cells using a glass-silicon anodic bonding technique and in situ chemical reaction between rubidium chloride and barium azide to produce Rb. Under controlled conditions...We describe the microfabrication of ^85Rb vapour cells using a glass-silicon anodic bonding technique and in situ chemical reaction between rubidium chloride and barium azide to produce Rb. Under controlled conditions, the pure metallic Rb drops and buffer gases were obtained in the cells with a few mm^3 internal volumes during the cell sealing process. At an ambient temperature of 90 ℃ the optical absorption resonance of ^85Rb D1 transition with proper broadening and the corresponding coherent population trapping （CPT） resonance, with a signal contrast of 1.5% and linewidth of about 1.7 kHz, have been detected. The sealing quality and the stability of the cells have also been demonstrated experimentally by using the helium leaking detection and the after-9-month optoelectronics measurement which shows a similar CPT signal as its original status. In addition, the physics package of chip-scale atomic clock （CSAC） based on the cell was realized. The measured frequency stability of the physics package can reach to 2.1 × 10^-10 at one second when the cell was heated to 100 ℃ which proved that the cell has the quality to be used in portable and battery-operated devices.展开更多
This presentation predicts the elastic properties of three-dimensional(3D)orthogonal woven composite(3DOWC)by finite element analysis based on micro/meso repeated unit cell(RUC)models.First,the properties of fiber yar...This presentation predicts the elastic properties of three-dimensional(3D)orthogonal woven composite(3DOWC)by finite element analysis based on micro/meso repeated unit cell(RUC)models.First,the properties of fiber yarn are obtained by analysis on a micro-scale RUC model assuming fibers in a hexagonal distribution pattern in the polymer matrix.Then a full thickness meso-scale RUC model including weft yarns,warp yarns,Z-yarns and pure resin zones is established and full stiffness matrix of the 3DOWC including the in-plane and flexural constants are predicted.For thick 3DOWC with large number of weft,warp layers,an alternative analysis method is proposed in which an inner meso-RUC and a surface meso-RUC are established,respectively.Then the properties of 3DOWC are deduced based on laminate theory and properties of the inner and surface layers.The predicted results by the above two alternative methods are in good experimental agreement.展开更多
Stem cells (SCs), the undifferentiated biological cells, have the infinite capacity to self-renew and the pluripotent ability to differentiate. SCs and their derived products offer great promise for biomedical applica...Stem cells (SCs), the undifferentiated biological cells, have the infinite capacity to self-renew and the pluripotent ability to differentiate. SCs and their derived products offer great promise for biomedical applications such as cell therapy, tissue engineering, regenerative medicine and drug screening. However, the clinical applications of SCs require a large amount of SCs with high quality and the number of SCs from their tissue resources is very limited. Large-scale expansion is needed to generate homogeneous SCs with good biological characteristics for clinical application. This necessitates a bioreactor system to provide controllable and stable conditions for stem cell (SC) culture. Traditional methods of bioreactor for maintenance and expansion of cells rely on two-dimensional (2-D) culture techniques, leading to loss self-renewal ability and differentiation capacity upon long-term culture. New approaches for SC expansion with bioreactor employ three-dimensional (3-D) cell growth to mimic their environment in vivo. In this review, we summarize the application of bioreactors in SC culture.展开更多
Effect of non-hydrostatic stress on X-ray diffraction in a diamond anvil cell （DAC） is studied. The pressure gradient in the sample chamber leads to the broadening of the diffraction peaks, which increase with the h...Effect of non-hydrostatic stress on X-ray diffraction in a diamond anvil cell （DAC） is studied. The pressure gradient in the sample chamber leads to the broadening of the diffraction peaks, which increase with the hkl index of the crystal. It is found that the difference between the determined d-spacing compressive ratio d/do and the real d-spacing compressive ratio dr/do is determined by the yield stress of the pressure transmitting media （if used） and the shear modulus of the sample. On the basis of the corrected experiment data of Mao et al. （MXB86）, which was used to calibrate the most widely used ruby fluorescence scale, a new relationship of ruby fluorescence pressure scale is corrected, i.e., P = （1904/9.827）[（1 ＋ △λ/λ0）9.827 - 1].展开更多
Objective:To judge the efficacies of neural stem cell(NSC)transplantation on functional recovery following contusion spinal cord injuries(SCIs).Data sources:Studies in which NSCs were transplanted into a clinically re...Objective:To judge the efficacies of neural stem cell(NSC)transplantation on functional recovery following contusion spinal cord injuries(SCIs).Data sources:Studies in which NSCs were transplanted into a clinically relevant,standardized rat model of contusion SCI were identified by searching the PubMed,Embase and Cochrane databases,and the extracted data were analyzed by Stata 14.0.Data selection:Inclusion criteria were that NSCs were used in in vivo animal studies to treat contusion SCIs and that behavioral assessment of locomotor functional recovery was performed using the Basso,Beattie,and Bresnahan lo-comotor rating scale.Exclusion criteria included a follow-up of less than 4 weeks and the lack of control groups.Outcome measures:The restoration of motor function was assessed by the Basso,Beattie,and Bresnahan locomotor rating scale.Results:We identified 1756 non-duplicated papers by searching the aforementioned electronic databases,and 30 full-text articles met the inclusion criteria.A total of 37 studies reported in the 30 articles were included in the meta-analysis.The meta-analysis results showed that transplanted NSCs could improve the motor function recovery of rats following contusion SCIs,to a moderate extent(pooled standardized mean difference(SMD)=0.73;95%confidence interval(CI):0.47–1.00;P<0.001).NSCs obtained from different donor species(rat:SMD=0.74;95%CI:0.36–1.13;human:SMD=0.78;95%CI:0.31–1.25),at different donor ages(fetal:SMD=0.67;95%CI:0.43–0.92;adult:SMD=0.86;95%CI:0.50–1.22)and from different origins(brain-derived:SMD=0.59;95%CI:0.27–0.91;spinal cord-derived:SMD=0.51;95%CI:0.22–0.79)had similar efficacies on improved functional recovery;however,adult induced pluripotent stem cell-derived NSCs showed no significant efficacies.Furthermore,the use of higher doses of transplanted NSCs or the administration of immunosuppressive agents did not promote better locomotor function recovery(SMD=0.45;95%CI:0.21–0.70).However,shorter periods between the contusion induction and the NSC transplantation showed slightly higher efficacies(acute:SMD=1.22;95%CI:0.81–1.63;subacute:SMD=0.75;95%CI:0.42–1.09).For chronic injuries,NSC implantation did not significantly improve functional recovery(SMD=0.25;95%CI:–0.16 to 0.65).Conclusion:NSC transplantation alone appears to be a positive yet limited method for the treatment of contusion SCIs.展开更多
Rhodioloside has been shown to protect cells from hypoxia injury,and bone marrow mesenchymal stem cells have a good effect on tissue repair.To study the effects of rhodioloside and bone marrow mesenchymal stem cells o...Rhodioloside has been shown to protect cells from hypoxia injury,and bone marrow mesenchymal stem cells have a good effect on tissue repair.To study the effects of rhodioloside and bone marrow mesenchymal stem cells on spinal cord injury,a rat model of spinal cord injury was established using the Infinite Horizons method.After establishing the model,the rats were randomly divided into five groups.Rats in the control group were intragastrically injected with phosphate buffered saline(PBS)(5μL).PBS was injected at 6 equidistant points around 5 mm from the injury site and at a depth of 5 mm.Rats in the rhodioloside group were intragastrically injected with rhodioloside(5 g/kg)and intramuscularly injected with PBS.Rats in the mesenchymal stem cell(MSC)group were intramuscularly injected with PBS and intramuscularly with MSCs(8×10^6/mL in a 50-μL cell suspension).Rats in the Ad-HIF-MSC group were intragastrically injected with PBS and intramuscularly injected with HIF-1 adenovirus-infected MSCs.Rats in the rhodioloside+Ad-HIF-MSC group were intramuscularly injected with MSCs infected with the HIF-1 adenovirus and intragastrically injected with rhodioloside.One week after treatment,exercise recovery was evaluated with a modified combined behavioral score scale.Hematoxylin-eosin staining and Pischingert’s methylene blue staining were used to detect any histological or pathological changes in spinal cord tissue.Levels of adenovirus IX and Sry mRNA were detected by real-time quantitative polymerase chain reaction and used to determine the number of adenovirus and mesenchymal stem cells that were transfected into the spinal cord.Immunohistochemical staining was applied to detect HIF-1 protein levels in the spinal cord.The results showed that:(1)compared with the other groups,the rhodioloside+Ad-HIF-MSC group exhibited the highest combined behavioral score(P<0.05),the most recovered tissue,and the greatest number of neurons,as indicated by Pischingert’s methylene blue staining.(2)Compared with the PBS group,HIF-1 protein expression was greater in the rhodioloside group(P<0.05).(3)Compared with the Ad-HIF-MSC group,Sry mRNA levels were higher in the rhodioloside+Ad-HIF-MSC group(P<0.05).These results confirm that rhodioloside combined with bone marrow mesenchymal stem cells can promote the recovery of spinal cord injury and activate the HIF-1 pathway to promote the survival of bone marrow mesenchymal stem cells and repair damaged neurons within spinal cord tissue.This experiment was approved by the Animal Ethics Committee of Gansu University of Traditional Chinese Medicine,China(approval No.2015KYLL029)in June 2015.展开更多
It was in flask optimization tests proved that 2% serum, pH 7.0, 5:10 000 inoculation concentration of infectious bursal disease virus (IBDV) and 108 hours cultivation for IBDV harvest after its inoculation were the o...It was in flask optimization tests proved that 2% serum, pH 7.0, 5:10 000 inoculation concentration of infectious bursal disease virus (IBDV) and 108 hours cultivation for IBDV harvest after its inoculation were the optimal conditions when IBDV was propagated on Vero cells. 250 ml self-made spinner bottle and 5 L stirring fermentor tests proved that IBDV could maintain higher liters for a long time and the highest liters of IBDV in a spinner bottle and a fermentor were 8.875 and 8.58 ( - lgTCID50/0.1 ml) respectively when IBDV was proliferated on Vero cells using 2 g/L microcarriers in a spinner bottle and a fermentor and was cultivated under the optimum conditions obtained from flask tests after Vero cells had developed a confluent monolayer on microcarriers, which were at least one titer higher than the highest titer in the traditional rolling bottle. All these results suggested that this technology could be applied to large scale production for IBDV.展开更多
基金Supported by In part by Florida State University start up fundFlorida State University Research Foundation GAP awardthe partial support from National Science Foundation,No.1342192
文摘Human pluripotent stem cells(hPSCs), including human embryonic stem cells and human induced pluripotent stem cells, are promising sources for hematopoietic cells due to their unlimited growth capacity and the pluripotency. Dendritic cells(DCs), the unique immune cells in the hematopoietic system, can be loaded with tumor specific antigen and used as vaccine for cancer immunotherapy. While autologous DCs from peripheral blood are limited in cell number, hPSC-derived DCs provide a novel alternative cell source which has the potential for large scale production. This review summarizes recent advances in differentiating hPSCs to DCs through the intermediate stage of hematopoietic stem cells. Step-wise growth factor induction has been used to derive DCs from hPSCs either in suspension cultureof embryoid bodies(EBs) or in co-culture with stromal cells. To fulfill the clinical potential of the DCs derived from hPSCs, the bioprocess needs to be scaled up to produce a large number of cells economically under tight quality control. This requires the development of novel bioreactor systems combining guided EB-based differentiation with engineered culture environment. Hence, recent progress in using bioreactors for hPSC lineage-specific differentiation is reviewed. In particular, the potential scale up strategies for the multistage DC differentiation and the effect of shear stress on hPSC differentiation in bioreactors are discussed in detail.
基金supported by the National Natural Science Foundation of China(6110602261574013)
文摘The influence of cells groupings factor to the performance of the cells groupings time-shift pilot scheme is researched for the multiple cells large scale antennas systems（LSAS）. The former researches have confirmed that the cells groupings time-shift pilots scheme is effective to reduce inter-cell interference, especially pilot contamination, which results from the pilot reuse in adjacent cells. However, they have not specified reasonable cells groupings factor, which plays a critical role in the general performance of the LSAS. Therefore, this problem is researched in details. The time for reverse-link data transmission will be compressed, when the groupings factor surpasses a certain range. Thus it is not always beneficial to increase the cells groupings factor without limitation. Furthermore,a reasonable cells groupings factor is deduced from the perspective of optimization to enhance the system performance. Simulations verify the proposed cell grouping factor.
文摘The catalyst layer （CL） of proton exchange mem-brane fuel cell （PEMFC） involves various particles and pores in meso-scale, which has an important effect on the mass, charge （proton and electron） and heat transport coupled with the electrochemical reactions. The coarse-grained molecular dynamics （CG-MD） method is employed as a meso-scale structure reconstruction technique to mimic the self-organization phenomena in the fabrication steps of a CL. The meso-scale structure obtained at the equilibrium state is further analyzed by molecular dynamic （MD） software to provide the necessary microscopic parameters for understanding of multi-scale and-physics processes in CLs. The primary pore size distribution （PSD） and active platinum （Pt） surface areas are also calculated and then compared with the experiments. In addition, we also highlight the implementation method to combine microscopic elementary kinetic reaction schemes with the CG-MD approaches to provide insight into the reactions in CLs. The concepts from CG modeling with particle hydrodynamics （SPH） and the problems on coupling of SPH with finite element modeling （FEM） methods are further outlined and discussed to understand the effects of the meso-scale transport phenomena in fuel cells.
基金Project supported by National 863/973 Plans Projects (Grant Nos. 2006AA04Z361,2006CB932402)NSFC (Grant No. 60971002)
文摘We describe the microfabrication of ^85Rb vapour cells using a glass-silicon anodic bonding technique and in situ chemical reaction between rubidium chloride and barium azide to produce Rb. Under controlled conditions, the pure metallic Rb drops and buffer gases were obtained in the cells with a few mm^3 internal volumes during the cell sealing process. At an ambient temperature of 90 ℃ the optical absorption resonance of ^85Rb D1 transition with proper broadening and the corresponding coherent population trapping （CPT） resonance, with a signal contrast of 1.5% and linewidth of about 1.7 kHz, have been detected. The sealing quality and the stability of the cells have also been demonstrated experimentally by using the helium leaking detection and the after-9-month optoelectronics measurement which shows a similar CPT signal as its original status. In addition, the physics package of chip-scale atomic clock （CSAC） based on the cell was realized. The measured frequency stability of the physics package can reach to 2.1 × 10^-10 at one second when the cell was heated to 100 ℃ which proved that the cell has the quality to be used in portable and battery-operated devices.
基金BASTRI Subtopic Research about Digital Sampler Technology of Body Structure Performance Study Based on Big Data Calculation Model,China(No.MIIT Civil aircraft special purpose MJ-2017-F-20)
文摘This presentation predicts the elastic properties of three-dimensional(3D)orthogonal woven composite(3DOWC)by finite element analysis based on micro/meso repeated unit cell(RUC)models.First,the properties of fiber yarn are obtained by analysis on a micro-scale RUC model assuming fibers in a hexagonal distribution pattern in the polymer matrix.Then a full thickness meso-scale RUC model including weft yarns,warp yarns,Z-yarns and pure resin zones is established and full stiffness matrix of the 3DOWC including the in-plane and flexural constants are predicted.For thick 3DOWC with large number of weft,warp layers,an alternative analysis method is proposed in which an inner meso-RUC and a surface meso-RUC are established,respectively.Then the properties of 3DOWC are deduced based on laminate theory and properties of the inner and surface layers.The predicted results by the above two alternative methods are in good experimental agreement.
文摘Stem cells (SCs), the undifferentiated biological cells, have the infinite capacity to self-renew and the pluripotent ability to differentiate. SCs and their derived products offer great promise for biomedical applications such as cell therapy, tissue engineering, regenerative medicine and drug screening. However, the clinical applications of SCs require a large amount of SCs with high quality and the number of SCs from their tissue resources is very limited. Large-scale expansion is needed to generate homogeneous SCs with good biological characteristics for clinical application. This necessitates a bioreactor system to provide controllable and stable conditions for stem cell (SC) culture. Traditional methods of bioreactor for maintenance and expansion of cells rely on two-dimensional (2-D) culture techniques, leading to loss self-renewal ability and differentiation capacity upon long-term culture. New approaches for SC expansion with bioreactor employ three-dimensional (3-D) cell growth to mimic their environment in vivo. In this review, we summarize the application of bioreactors in SC culture.
基金Project supported by the Defense Industrial Technology Development Program of China (Grant No. B1520110001)the National Natural Science Foundation of China (Grant No. 10874158)the Project of China Academy of Engineering Physics (Grant Nos. 2010A0101001 and 2008A0101001)
文摘Effect of non-hydrostatic stress on X-ray diffraction in a diamond anvil cell （DAC） is studied. The pressure gradient in the sample chamber leads to the broadening of the diffraction peaks, which increase with the hkl index of the crystal. It is found that the difference between the determined d-spacing compressive ratio d/do and the real d-spacing compressive ratio dr/do is determined by the yield stress of the pressure transmitting media （if used） and the shear modulus of the sample. On the basis of the corrected experiment data of Mao et al. （MXB86）, which was used to calibrate the most widely used ruby fluorescence scale, a new relationship of ruby fluorescence pressure scale is corrected, i.e., P = （1904/9.827）[（1 ＋ △λ/λ0）9.827 - 1].
基金supported by the National Natural Science Foundation of China,No.81171147“Key Medical Talents of Qiangwei Project” Research Foundation of Health Department of Jiangsu Province of China,No.ZDRCA2016010+1 种基金“Xingwei Project” Key Personal Medical Research Foundation of Health Department of Jiangsu Province of China,No.RC201156Jiangsu Provincial Key Discipline of Medicine of China,No.XK201117(all to LXL)
文摘Objective:To judge the efficacies of neural stem cell(NSC)transplantation on functional recovery following contusion spinal cord injuries(SCIs).Data sources:Studies in which NSCs were transplanted into a clinically relevant,standardized rat model of contusion SCI were identified by searching the PubMed,Embase and Cochrane databases,and the extracted data were analyzed by Stata 14.0.Data selection:Inclusion criteria were that NSCs were used in in vivo animal studies to treat contusion SCIs and that behavioral assessment of locomotor functional recovery was performed using the Basso,Beattie,and Bresnahan lo-comotor rating scale.Exclusion criteria included a follow-up of less than 4 weeks and the lack of control groups.Outcome measures:The restoration of motor function was assessed by the Basso,Beattie,and Bresnahan locomotor rating scale.Results:We identified 1756 non-duplicated papers by searching the aforementioned electronic databases,and 30 full-text articles met the inclusion criteria.A total of 37 studies reported in the 30 articles were included in the meta-analysis.The meta-analysis results showed that transplanted NSCs could improve the motor function recovery of rats following contusion SCIs,to a moderate extent(pooled standardized mean difference(SMD)=0.73;95%confidence interval(CI):0.47–1.00;P<0.001).NSCs obtained from different donor species(rat:SMD=0.74;95%CI:0.36–1.13;human:SMD=0.78;95%CI:0.31–1.25),at different donor ages(fetal:SMD=0.67;95%CI:0.43–0.92;adult:SMD=0.86;95%CI:0.50–1.22)and from different origins(brain-derived:SMD=0.59;95%CI:0.27–0.91;spinal cord-derived:SMD=0.51;95%CI:0.22–0.79)had similar efficacies on improved functional recovery;however,adult induced pluripotent stem cell-derived NSCs showed no significant efficacies.Furthermore,the use of higher doses of transplanted NSCs or the administration of immunosuppressive agents did not promote better locomotor function recovery(SMD=0.45;95%CI:0.21–0.70).However,shorter periods between the contusion induction and the NSC transplantation showed slightly higher efficacies(acute:SMD=1.22;95%CI:0.81–1.63;subacute:SMD=0.75;95%CI:0.42–1.09).For chronic injuries,NSC implantation did not significantly improve functional recovery(SMD=0.25;95%CI:–0.16 to 0.65).Conclusion:NSC transplantation alone appears to be a positive yet limited method for the treatment of contusion SCIs.
基金supported by the National High Technology Research and Development Program of China (863 Program), No. 2015CB755400 (to XQH)
文摘Rhodioloside has been shown to protect cells from hypoxia injury,and bone marrow mesenchymal stem cells have a good effect on tissue repair.To study the effects of rhodioloside and bone marrow mesenchymal stem cells on spinal cord injury,a rat model of spinal cord injury was established using the Infinite Horizons method.After establishing the model,the rats were randomly divided into five groups.Rats in the control group were intragastrically injected with phosphate buffered saline(PBS)(5μL).PBS was injected at 6 equidistant points around 5 mm from the injury site and at a depth of 5 mm.Rats in the rhodioloside group were intragastrically injected with rhodioloside(5 g/kg)and intramuscularly injected with PBS.Rats in the mesenchymal stem cell(MSC)group were intramuscularly injected with PBS and intramuscularly with MSCs(8×10^6/mL in a 50-μL cell suspension).Rats in the Ad-HIF-MSC group were intragastrically injected with PBS and intramuscularly injected with HIF-1 adenovirus-infected MSCs.Rats in the rhodioloside+Ad-HIF-MSC group were intramuscularly injected with MSCs infected with the HIF-1 adenovirus and intragastrically injected with rhodioloside.One week after treatment,exercise recovery was evaluated with a modified combined behavioral score scale.Hematoxylin-eosin staining and Pischingert’s methylene blue staining were used to detect any histological or pathological changes in spinal cord tissue.Levels of adenovirus IX and Sry mRNA were detected by real-time quantitative polymerase chain reaction and used to determine the number of adenovirus and mesenchymal stem cells that were transfected into the spinal cord.Immunohistochemical staining was applied to detect HIF-1 protein levels in the spinal cord.The results showed that:(1)compared with the other groups,the rhodioloside+Ad-HIF-MSC group exhibited the highest combined behavioral score(P<0.05),the most recovered tissue,and the greatest number of neurons,as indicated by Pischingert’s methylene blue staining.(2)Compared with the PBS group,HIF-1 protein expression was greater in the rhodioloside group(P<0.05).(3)Compared with the Ad-HIF-MSC group,Sry mRNA levels were higher in the rhodioloside+Ad-HIF-MSC group(P<0.05).These results confirm that rhodioloside combined with bone marrow mesenchymal stem cells can promote the recovery of spinal cord injury and activate the HIF-1 pathway to promote the survival of bone marrow mesenchymal stem cells and repair damaged neurons within spinal cord tissue.This experiment was approved by the Animal Ethics Committee of Gansu University of Traditional Chinese Medicine,China(approval No.2015KYLL029)in June 2015.
文摘It was in flask optimization tests proved that 2% serum, pH 7.0, 5:10 000 inoculation concentration of infectious bursal disease virus (IBDV) and 108 hours cultivation for IBDV harvest after its inoculation were the optimal conditions when IBDV was propagated on Vero cells. 250 ml self-made spinner bottle and 5 L stirring fermentor tests proved that IBDV could maintain higher liters for a long time and the highest liters of IBDV in a spinner bottle and a fermentor were 8.875 and 8.58 ( - lgTCID50/0.1 ml) respectively when IBDV was proliferated on Vero cells using 2 g/L microcarriers in a spinner bottle and a fermentor and was cultivated under the optimum conditions obtained from flask tests after Vero cells had developed a confluent monolayer on microcarriers, which were at least one titer higher than the highest titer in the traditional rolling bottle. All these results suggested that this technology could be applied to large scale production for IBDV.
