Müller glia,as prominent glial cells within the retina,plays a significant role in maintaining retinal homeostasis in both healthy and diseased states.In lower vertebrates like zebrafish,these cells assume respon...Müller glia,as prominent glial cells within the retina,plays a significant role in maintaining retinal homeostasis in both healthy and diseased states.In lower vertebrates like zebrafish,these cells assume responsibility for spontaneous retinal regeneration,wherein endogenous Müller glia undergo proliferation,transform into Müller glia-derived progenitor cells,and subsequently regenerate the entire retina with restored functionality.Conversely,Müller glia in the mouse and human retina exhibit limited neural reprogramming.Müller glia reprogramming is thus a promising strategy for treating neurodegenerative ocular disorders.Müller glia reprogramming in mice has been accomplished with remarkable success,through various technologies.Advancements in molecular,genetic,epigenetic,morphological,and physiological evaluations have made it easier to document and investigate the Müller glia programming process in mice.Nevertheless,there remain issues that hinder improving reprogramming efficiency and maturity.Thus,understanding the reprogramming mechanism is crucial toward exploring factors that will improve Müller glia reprogramming efficiency,and for developing novel Müller glia reprogramming strategies.This review describes recent progress in relatively successful Müller glia reprogramming strategies.It also provides a basis for developing new Müller glia reprogramming strategies in mice,including epigenetic remodeling,metabolic modulation,immune regulation,chemical small-molecules regulation,extracellular matrix remodeling,and cell-cell fusion,to achieve Müller glia reprogramming in mice.展开更多
The phyAm gene encoding acid phytase and optimized neutral phytase phyCs gene were inserted into expression vector pPIC9K in correct orientation and transformed into Pichia pastoris in order to expand the pH profile o...The phyAm gene encoding acid phytase and optimized neutral phytase phyCs gene were inserted into expression vector pPIC9K in correct orientation and transformed into Pichia pastoris in order to expand the pH profile of phytase and decrease the cost of production. The fusion phytase phyAm-phyCs gene was successfully overexpressed in P. pastoris as an active and ex-tracellular phytase. The yield of total extracellular fusion phytase activity is (25.4±0.53) U/ml at the flask scale and (159.1±2.92) U/ml for high cell-density fermentation, respectively. Purified fusion phytase exhibits an optimal temperature at 55 °C and an optimal pH at 5.5~6.0 and its relative activity remains at a relatively high level of above 70% in the range of pH 2.0 to 7.0. About 51% to 63% of its original activity remains after incubation at 75 °C to 95 °C for 10 min. Due to heavy glycosylation, the expressed fusion phytase shows a broad and diffuse band in SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis). After deglycosylation by endoglycosidase H (EndoHf), the enzyme has an apparent molecular size of 95 kDa. The characterization of the fusion phytase was compared with those of phyCs and phyAm.展开更多
Large numbers of precision fusion excitation functions were fitted in the literature using the nucleus-nucleus interaction potential having the Woods-Saxon shape. The diffuseness of this potential fusion ranges from 0...Large numbers of precision fusion excitation functions were fitted in the literature using the nucleus-nucleus interaction potential having the Woods-Saxon shape. The diffuseness of this potential fusion ranges from 0.75 to 1.5 fm. This is much larger than the value of 0.65 fm required by the elastic scattering data. Trying to resolve this contradiction we develop the dissipative trajectory model based on the density-dependent M3Y NN-forces folded with the nuclear matter distribution. Resulting potential possesses the normal diffuseness about 0.65 fm. With this potential we reach the agreement with the data for 16O+208Pb, 28Si+208Pb, 32S+208Pb reactions within 5%.展开更多
Introduction: Since the earliest description of spinal fusion in 1911 and later by Dr. Fred H. Albee, it has become one of the most commonly performed procedures by orthopedist and neurosurgeons. The spinal fusion is ...Introduction: Since the earliest description of spinal fusion in 1911 and later by Dr. Fred H. Albee, it has become one of the most commonly performed procedures by orthopedist and neurosurgeons. The spinal fusion is now used to treat a variety of indications, such as traumatic injuries, deformities, primary and secondary tumors, infections and degenerative conditions of the spine. The risk of iatrogenic injury during traditional anterior, posterior, and transforaminal open fusion surgery is significant. The axial lumbar interbody fusion (Axia-LIF) is a minimal invasive technique which uses the retroperitoneumpresacral anatomical corridor to fuse the lumbar vertebral bodies L4-L5-S1 avoiding manipulation of the annular ligament, paravertebral muscles and facet joints. Methods: In this retrospective series, we report all the cases made in the Centro Medico Naval in México City in two years. A total of eleven patients with degenerative disc disease and spondylolisthesis underwent Axia-LIF one or two level systems with a 36 months clinical and radiographic follow-up. The outcomes included Oswestry Disability Index (ODI) score and leg/back pain severity. Radiographic outcome was evaluated with dynamics and orthogonal x-ray, as well as lumbosacral tomography scan to evaluate fusion status. Results: Nine patients underwent Axia-LIF one level system (L5-S1) and the rest two levels system (L4-S1). Ten patients were fixated with transpedicular percutaneous screws and one with facets joints screws. No intraoperative complications were reported. The mean back pain severity improved 57% in 12 months, and the mean leg pain severity improved 50% in the same time (P < 0.001). Mean ODI scores improved 58%, from 60% ± 16% at baseline to 25% ± 8% at twelve months (P < 0.001). At one year, a patient developed pseudoarthrosis that required posterolateral arthrodesis with transpedicular percutaneous screws. At 36 months monitoring, 100% patients presented a total interbody fusion in the tomography scans. At final follow-up, mean ODI score improved 73% (16% ± 5%;P < 0.001). Conclusion: The Axial Lumbar Interbody Fusion has demonstrated to be a safe treatment for the degenerative disc disease L5-S1 and L4-S1. The patients who underwent one or two level Axia-LIF showed an improvement in ODI and back/leg pain severity scores, with no intraoperative complications. The use of this technique and its indications are still in controversy;nevertheless, its use has increased as for pathologies such as spondylitis, scoliosis, patients with residual pain with previous surgeries. We recommended complementary pedicular fixation to avoid complications and improved interbody fusion.展开更多
Influenza A virus(IAV) commandeers numerous host cellular factors for successful replication. However, very few host factors have been revealed to be involved in the fusion of viral envelope and late endosomal membran...Influenza A virus(IAV) commandeers numerous host cellular factors for successful replication. However, very few host factors have been revealed to be involved in the fusion of viral envelope and late endosomal membranes. In this study, we identified cation-dependent mannose-6-phosphate receptor(M6PR) as a crucial host factor for the replication of IAV. We found that siRNA knockdown of M6PR expression significantly reduced the growth titers of different subtypes of IAV, and that the inhibitory effect of M6PR siRNA treatment on IAV growth was overcome by the complement of exogenously expressed M6PR. When A549 cells were treated with siRNA targeting M6PR,the nuclear accumulation of viral nucleoprotein(NP) was dramatically inhibited at early timepoints post-infection, indicating that M6PR engages in the early stage of the IAV replication cycle. By investigating the role of M6PR in the individual entry and post-entry steps of IAV replication, we found that the downregulation of M6PR expression had no effect on attachment, internalization, early endosome trafficking,or late endosome acidification. However, we found that M6PR expression was critical for the fusion of viral envelope and late endosomal membranes. Of note, M6PR interacted with the hemagglutinin(HA) protein of IAV, and further studies showed that the lumenal domain of M6PR and the ectodomain of HA2 mediated the interaction and directly promoted the fusion of the viral and late endosomal membranes,thereby facilitating IAV replication. Together, our findings highlight the importance of the M6PR–HA interaction in the fusion of viral and late endosomal membranes during IAV replication.展开更多
基金supported by the National Natural Science Foundation of China,No.31930068National Key Research and Development Program of China,Nos.2018YFA0107302 and 2021YFA1101203(all to HX).
文摘Müller glia,as prominent glial cells within the retina,plays a significant role in maintaining retinal homeostasis in both healthy and diseased states.In lower vertebrates like zebrafish,these cells assume responsibility for spontaneous retinal regeneration,wherein endogenous Müller glia undergo proliferation,transform into Müller glia-derived progenitor cells,and subsequently regenerate the entire retina with restored functionality.Conversely,Müller glia in the mouse and human retina exhibit limited neural reprogramming.Müller glia reprogramming is thus a promising strategy for treating neurodegenerative ocular disorders.Müller glia reprogramming in mice has been accomplished with remarkable success,through various technologies.Advancements in molecular,genetic,epigenetic,morphological,and physiological evaluations have made it easier to document and investigate the Müller glia programming process in mice.Nevertheless,there remain issues that hinder improving reprogramming efficiency and maturity.Thus,understanding the reprogramming mechanism is crucial toward exploring factors that will improve Müller glia reprogramming efficiency,and for developing novel Müller glia reprogramming strategies.This review describes recent progress in relatively successful Müller glia reprogramming strategies.It also provides a basis for developing new Müller glia reprogramming strategies in mice,including epigenetic remodeling,metabolic modulation,immune regulation,chemical small-molecules regulation,extracellular matrix remodeling,and cell-cell fusion,to achieve Müller glia reprogramming in mice.
基金the National Key Technologies R & D Program of China during the 10th Five-Year Plan Period (No. 2002BA514A-12)the Education Department of Sichuan Province (No. 2006B014)the Innovative Fund for Distinguished Young Scholars of Sichuan Agricultural University, China
文摘The phyAm gene encoding acid phytase and optimized neutral phytase phyCs gene were inserted into expression vector pPIC9K in correct orientation and transformed into Pichia pastoris in order to expand the pH profile of phytase and decrease the cost of production. The fusion phytase phyAm-phyCs gene was successfully overexpressed in P. pastoris as an active and ex-tracellular phytase. The yield of total extracellular fusion phytase activity is (25.4±0.53) U/ml at the flask scale and (159.1±2.92) U/ml for high cell-density fermentation, respectively. Purified fusion phytase exhibits an optimal temperature at 55 °C and an optimal pH at 5.5~6.0 and its relative activity remains at a relatively high level of above 70% in the range of pH 2.0 to 7.0. About 51% to 63% of its original activity remains after incubation at 75 °C to 95 °C for 10 min. Due to heavy glycosylation, the expressed fusion phytase shows a broad and diffuse band in SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis). After deglycosylation by endoglycosidase H (EndoHf), the enzyme has an apparent molecular size of 95 kDa. The characterization of the fusion phytase was compared with those of phyCs and phyAm.
文摘Large numbers of precision fusion excitation functions were fitted in the literature using the nucleus-nucleus interaction potential having the Woods-Saxon shape. The diffuseness of this potential fusion ranges from 0.75 to 1.5 fm. This is much larger than the value of 0.65 fm required by the elastic scattering data. Trying to resolve this contradiction we develop the dissipative trajectory model based on the density-dependent M3Y NN-forces folded with the nuclear matter distribution. Resulting potential possesses the normal diffuseness about 0.65 fm. With this potential we reach the agreement with the data for 16O+208Pb, 28Si+208Pb, 32S+208Pb reactions within 5%.
文摘Introduction: Since the earliest description of spinal fusion in 1911 and later by Dr. Fred H. Albee, it has become one of the most commonly performed procedures by orthopedist and neurosurgeons. The spinal fusion is now used to treat a variety of indications, such as traumatic injuries, deformities, primary and secondary tumors, infections and degenerative conditions of the spine. The risk of iatrogenic injury during traditional anterior, posterior, and transforaminal open fusion surgery is significant. The axial lumbar interbody fusion (Axia-LIF) is a minimal invasive technique which uses the retroperitoneumpresacral anatomical corridor to fuse the lumbar vertebral bodies L4-L5-S1 avoiding manipulation of the annular ligament, paravertebral muscles and facet joints. Methods: In this retrospective series, we report all the cases made in the Centro Medico Naval in México City in two years. A total of eleven patients with degenerative disc disease and spondylolisthesis underwent Axia-LIF one or two level systems with a 36 months clinical and radiographic follow-up. The outcomes included Oswestry Disability Index (ODI) score and leg/back pain severity. Radiographic outcome was evaluated with dynamics and orthogonal x-ray, as well as lumbosacral tomography scan to evaluate fusion status. Results: Nine patients underwent Axia-LIF one level system (L5-S1) and the rest two levels system (L4-S1). Ten patients were fixated with transpedicular percutaneous screws and one with facets joints screws. No intraoperative complications were reported. The mean back pain severity improved 57% in 12 months, and the mean leg pain severity improved 50% in the same time (P < 0.001). Mean ODI scores improved 58%, from 60% ± 16% at baseline to 25% ± 8% at twelve months (P < 0.001). At one year, a patient developed pseudoarthrosis that required posterolateral arthrodesis with transpedicular percutaneous screws. At 36 months monitoring, 100% patients presented a total interbody fusion in the tomography scans. At final follow-up, mean ODI score improved 73% (16% ± 5%;P < 0.001). Conclusion: The Axial Lumbar Interbody Fusion has demonstrated to be a safe treatment for the degenerative disc disease L5-S1 and L4-S1. The patients who underwent one or two level Axia-LIF showed an improvement in ODI and back/leg pain severity scores, with no intraoperative complications. The use of this technique and its indications are still in controversy;nevertheless, its use has increased as for pathologies such as spondylitis, scoliosis, patients with residual pain with previous surgeries. We recommended complementary pedicular fixation to avoid complications and improved interbody fusion.
文摘针对传统断路器机械特性评估方法的不足,构建了基于稀疏表示和M-ELM(Memetic-Extreme Learning Machine)的断路器故障诊断模型,该模型首先利用稀疏表示方法跟踪断路器分(合)闸过程中与动触头同步运动的连杆或主轴上辅助标志物运动的轨迹;然后,根据该轨迹获取断路器操动机构的行程时间曲线并据此计算断路器的分(合)闸速度等各种机械特性参数;最后,将动触头运动机械特性参数作为M-ELM的输入进行断路器的故障诊断。通过对12 k V真空断路器的测试实验证明了该模型诊断断路器状态的有效性和优越性。
基金supported by the National Natural Science Foundation of China(32192453,32172847)the National Key Research and Development Program of China(2021YFD1800204)+1 种基金the Laboratory of Lingnan Modern Agriculture Project(NT2021007)the earmarked fund for CARS-41。
文摘Influenza A virus(IAV) commandeers numerous host cellular factors for successful replication. However, very few host factors have been revealed to be involved in the fusion of viral envelope and late endosomal membranes. In this study, we identified cation-dependent mannose-6-phosphate receptor(M6PR) as a crucial host factor for the replication of IAV. We found that siRNA knockdown of M6PR expression significantly reduced the growth titers of different subtypes of IAV, and that the inhibitory effect of M6PR siRNA treatment on IAV growth was overcome by the complement of exogenously expressed M6PR. When A549 cells were treated with siRNA targeting M6PR,the nuclear accumulation of viral nucleoprotein(NP) was dramatically inhibited at early timepoints post-infection, indicating that M6PR engages in the early stage of the IAV replication cycle. By investigating the role of M6PR in the individual entry and post-entry steps of IAV replication, we found that the downregulation of M6PR expression had no effect on attachment, internalization, early endosome trafficking,or late endosome acidification. However, we found that M6PR expression was critical for the fusion of viral envelope and late endosomal membranes. Of note, M6PR interacted with the hemagglutinin(HA) protein of IAV, and further studies showed that the lumenal domain of M6PR and the ectodomain of HA2 mediated the interaction and directly promoted the fusion of the viral and late endosomal membranes,thereby facilitating IAV replication. Together, our findings highlight the importance of the M6PR–HA interaction in the fusion of viral and late endosomal membranes during IAV replication.
