Grain size is one of the most important factors affecting rice grain quality and yield,and attracts great attention from molecular biologists and breeders.In this study,we engineered a CRISPR/Cas9 system targeting the...Grain size is one of the most important factors affecting rice grain quality and yield,and attracts great attention from molecular biologists and breeders.In this study,we engineered a CRISPR/Cas9 system targeting the miR396 recognition site of the rice GS2 gene,which encodes growth-regulating factor 4(OsGRF4)and regulates multiple agronomic traits including grain size,grain quality,nitrogen use efficiency,abiotic stress response,and seed shattering.In contrast to most previous genome editing efforts in which indel mutations were chosen to obtain null mutants,a mutant named GS2^(E) carrying an in-frame 6-bp deletion and 1-bp substitution within the miR396-targeted sequence was identified.GS2^(E) plants showed increased expression of GS2 in consistent with impaired repression by miR396.As expected,the gain-of-function GS2^(E) mutant exhibited multiple beneficial traits including increased grain size and yield and bigger grain length/width ratio.Thousand grain weight and grain yield per plant of GS2^(E) plants were increased by 23.5%and 10.4%,respectively.These improved traits were passed to hybrids in a semidominant way,suggesting that the new GS2^(E) allele has great potential in rice improvement.Taken together,we report new GS2 germplasm and describe a novel gene-editing strategy that can be widely employed to improve grain size and yield in rice.This trait-improvement strategy could be applied to other genes containing miRNA target sites,in particular the conserved miR396-GRF/GIF module that governs plant growth,development and environmental response.展开更多
Hypoxia-Ischemia Brain Damage(HIBD)results in a widespread neuronal damage and permanent brain tissue injury due to the severe reactive oxygen species(ROS)boost induced neuroinflammation.In this work,a K doped N-C bas...Hypoxia-Ischemia Brain Damage(HIBD)results in a widespread neuronal damage and permanent brain tissue injury due to the severe reactive oxygen species(ROS)boost induced neuroinflammation.In this work,a K doped N-C based nanozyme was fabricated for scavenging ROS.KNC could impair ROS production and M1 polarization in microglia.Beneficial from these contents,the brain damage was mitigated in HIBD rats,which was proved by the increased regional blood flow,decreased pro-inflammatory microglia and astrocyte activation.The learning and memory capabilities were restored after applying with KNC post HIBD,which was ascribed to the diminished HI-induced dendritic spine loss in hippocampal regions.RNA-seq revealed that decreased ALOX12 expression is one of the clues of neuronal protection.KNC could combine with ALOX12 and further inhibit the lipid peroxi-dation.These two clues obtain KNC with superior ability of inhibition of ROS boost induced brain damage post HIBD.This nanozyme provided a potential strategies and new idea of HIBD therapy.展开更多
It is important to eliminate lipopolysaccharide(LPS)along with killing bacteria in periprosthetic joint infection(PJI)therapy for promoting bone repair due to its effect to regulate macrophages response.Although natur...It is important to eliminate lipopolysaccharide(LPS)along with killing bacteria in periprosthetic joint infection(PJI)therapy for promoting bone repair due to its effect to regulate macrophages response.Although natural antimicrobial peptides(AMPs)offer a good solution,the unknown toxicity,high cost and exogenetic immune response hamper their applications in clinic.In this work,we fabricated a nanowire-like composite material,named P@C,by combining chitosan and puerarin via solid-phase reaction,which can finely mimic the bio-functions of AMPs.Chitosan,serving as the bacteria membrane puncture agent,and puerarin,serving as the LPS target agent,synergistically destroy the bacterial membrane structure and inhibit its recovery,thus endowing P@C with good antibacterial property.In addition,P@C possesses good osteoimmunomodulation due to its ability of LPS elimination and macrophage differentiation modulation.The in vivo results show that P@C can inhibit the LPS induced bone destruction in the Escherichia coli infected rat.P@C exhibits superior bone regeneration in Escherichia coli infected rat due to the comprehensive functions of its superior antibacterial property,and its ability of LPS elimination and immunomodulation.P@C can well mimic the functions of AMPs,which provides a novel and effective method for treating the PJI in clinic.展开更多
The adverse immune response mediated by macrophages is one of the main factors that are prone to lead poor osseointegration of polyetheretherketone(PEEK)implants in clinic.Hence,endowing PEEK with immunomodulatory abi...The adverse immune response mediated by macrophages is one of the main factors that are prone to lead poor osseointegration of polyetheretherketone(PEEK)implants in clinic.Hence,endowing PEEK with immunomodulatory ability to avoid the adverse immune response becomes a promising strategy to promote bone repair.In this work,sulfonation and hydrothermal treatment were used to fabricate a 3D porous surface on PEEK and hydroxyapatite(HA)composited PEEK.The HA composited PEEK with 3D porous surface inhibited macrophages polarizing to M1 phenotype and downregulated inducible nitric oxide synthase protein expression,which led to a nitric oxide concentration reduction in culture medium of mouse bone marrow mesenchymal stem cells(mBMSCs)under coculture condition.The decrease of nitric oxide concentration could help to increase bone formation-related OSX and ALP genes expressions and decrease bone resorption-related MMP-9 and MMP-13 genes expressions via cAMP–PKA–RUNX2 pathway in mBMSCs.In summary,the HA composited PEEK with 3D porous surface has the potential to promote osteogenesis of PEEK through immunomodulation,which provides a promising strategy to improve the bone repair ability of PEEK.展开更多
Ds-block elements have been gaining increasing attention in the field of biomaterials modification,owing to their excellent biological properties,such as antibiosis,osteogenesis,etc.However,their function mechanisms a...Ds-block elements have been gaining increasing attention in the field of biomaterials modification,owing to their excellent biological properties,such as antibiosis,osteogenesis,etc.However,their function mechanisms are not well understood and conflicting conclusions were drawn by previous studies on this issue,which are mainly resulted from the inconsistent experimental conditions.In this work,three most widely used ds-block elements,copper,zinc,and silver were introduced on titanium substrate by plasma immersion ion implantation method to investigate the rule of ds-block elements in the immune responses.Results showed that the implanted samples could decrease the inflammatory responses compared with Ti sample.The trend of anti-inflammatory effects of macrophages on samples was in correlation with cellular ROS levels,which was induced by the implanted biomaterials and positively correlated with the number of valence electrons of ds-block elements.The co-culture experiments of macrophages and bone marrow mesenchymal stem cells showed that these two kinds of cells could enhance the anti-inflammation and osteogenesis of samples by the paracrine manner of PGE2.In general,in their steady states on titanium substrate(Cu2+,Zn2+,Ag),the ds-block elements with more valence electrons exhibit better anti-inflammatory and osteogenic effects.Moreover,molecular biology experiments indicate that the PGE2-related signaling pathway may contribute to the desired immunoregulation and osteoinduction capability of ds-block elements.These findings suggest a correlation between the number of valence electrons of ds-block elements and the relevant biological responses,which provides new insight into the selection of implanted ions and surface design of biomaterials.展开更多
Infection and poor tissue repair are the key causes of percutaneous implantation failure. However, there is a lackof effective strategies to cope with due to its high requirements of sterilization, soft tissue healing...Infection and poor tissue repair are the key causes of percutaneous implantation failure. However, there is a lackof effective strategies to cope with due to its high requirements of sterilization, soft tissue healing, andosseointegration. In this work, L-arginine (L-Arg) was loaded onto a sulfonated polyetheretherketone (PEEK)surface to solve this issue. Under the infection condition, nitric oxide (NO) and reactive oxygen species (ROS) areproduced through catalyzing L-Arg by inducible nitric oxide synthase (iNOS) and thus play a role in bacteriasterilization. Under the tissue repair condition, L-Arg is catalyzed to ornithine by Arginase-1 (Arg-1), whichpromotes the proliferation and collagen secretion of L929 and rBMSCs. Notably, L-Arg loading samples couldpolarize macrophages to M1 and M2 in infection and tissue repair conditions, respectively. The results in vivoshow that the L-Arg loading samples could enhance infected soft tissue sealing and bone regeneration. Insummary, L-Arg loading sulfonated PEEK could polarize macrophage through metabolic reprogramming,providing multi-functions of antibacterial abilities, soft tissue repair, and bone regeneration, which gives a newidea to design percutaneous implantation materials.展开更多
Polyetheretherketone(PEEK)has been used as an implant material because it has similar mechani-cal properties to natural bone.However,inferior osseointegration and bioinertness hamper the clinical application of PEEK.I...Polyetheretherketone(PEEK)has been used as an implant material because it has similar mechani-cal properties to natural bone.However,inferior osseointegration and bioinertness hamper the clinical application of PEEK.In this study,the surfaces of sulfonated three-dimensional(3D)PEEK porous structures were loaded with different concentrations of strontium ranelate,a compound commonly used in the treatment or prevention of osteoporosis by promoting bone formation and inhibiting bone resorption.Field-emission scanning electron microscopy was used to characterize the topography of the structures,elemental carbon,oxygen and strontium contents were mea-sured by X-ray photoelectron spectroscopy,and surface zeta potentials and water-contact angle were also measured.The results indicated that strontium ranelate was successfully loaded onto the 3D porous structures.In vitro cellular results showed that strontium ranelate-treated sulfonated PEEK(SP-SR)strengthened the adhesion of MC3T3-E1 cells.The activity of alkaline phosphatase,collagen secretion and extracellular matrix mineralization deposition of MC3T3-E1 cells were also improved on the surface of SP-SR.These results indicate that SP-SR could serve a new implant candidate for surgical treatment.展开更多
基金supported by the National Key Research and Development Program of China(2016YFD0102000)“Breeding of Major New Varieties of Main Grain Crops”Program(2020ABA016)from Department of Science and Technology of Hubei Province.
文摘Grain size is one of the most important factors affecting rice grain quality and yield,and attracts great attention from molecular biologists and breeders.In this study,we engineered a CRISPR/Cas9 system targeting the miR396 recognition site of the rice GS2 gene,which encodes growth-regulating factor 4(OsGRF4)and regulates multiple agronomic traits including grain size,grain quality,nitrogen use efficiency,abiotic stress response,and seed shattering.In contrast to most previous genome editing efforts in which indel mutations were chosen to obtain null mutants,a mutant named GS2^(E) carrying an in-frame 6-bp deletion and 1-bp substitution within the miR396-targeted sequence was identified.GS2^(E) plants showed increased expression of GS2 in consistent with impaired repression by miR396.As expected,the gain-of-function GS2^(E) mutant exhibited multiple beneficial traits including increased grain size and yield and bigger grain length/width ratio.Thousand grain weight and grain yield per plant of GS2^(E) plants were increased by 23.5%and 10.4%,respectively.These improved traits were passed to hybrids in a semidominant way,suggesting that the new GS2^(E) allele has great potential in rice improvement.Taken together,we report new GS2 germplasm and describe a novel gene-editing strategy that can be widely employed to improve grain size and yield in rice.This trait-improvement strategy could be applied to other genes containing miRNA target sites,in particular the conserved miR396-GRF/GIF module that governs plant growth,development and environmental response.
基金Financial support from the Science and Technology Commission of Shanghai Municipality(24CL2900700)the Fundamental Research Funds for The Central Universities(YG2023ZD29)+5 种基金the National Natural Science Foundation of China(32371397,U21A20100)the Funda-mental Research Funds for The Central Universities(2232023A-10)the Research Fund of Shanghai Tongren Hospital,Shanghai Jiaotong Uni-versity School of Medicine(2023DHYGJC-YBA01)the Laboratory Open Fund of Key Technology and Materials in Minimally Invasive Spine Surgery(2024JZWC-ZDB03)Natural Science Foundation of Shanghai(24ZR1463600)the Talent project of Shanghai Tongren Hospital(TRKYRC-xx02)are acknowledged.
文摘Hypoxia-Ischemia Brain Damage(HIBD)results in a widespread neuronal damage and permanent brain tissue injury due to the severe reactive oxygen species(ROS)boost induced neuroinflammation.In this work,a K doped N-C based nanozyme was fabricated for scavenging ROS.KNC could impair ROS production and M1 polarization in microglia.Beneficial from these contents,the brain damage was mitigated in HIBD rats,which was proved by the increased regional blood flow,decreased pro-inflammatory microglia and astrocyte activation.The learning and memory capabilities were restored after applying with KNC post HIBD,which was ascribed to the diminished HI-induced dendritic spine loss in hippocampal regions.RNA-seq revealed that decreased ALOX12 expression is one of the clues of neuronal protection.KNC could combine with ALOX12 and further inhibit the lipid peroxi-dation.These two clues obtain KNC with superior ability of inhibition of ROS boost induced brain damage post HIBD.This nanozyme provided a potential strategies and new idea of HIBD therapy.
基金National Natural Science Foundation of China(U21A20100,32000938)Science and Technology Commission of Shanghai Municipality,China(19JC1415500,20ZR1465000)+1 种基金Shenzhen Science and Technology Funding(JCYJ20210324120009026)S&T Innovation 2025 Major Special Program of Ningbo(2018B10040)are acknowledged.
文摘It is important to eliminate lipopolysaccharide(LPS)along with killing bacteria in periprosthetic joint infection(PJI)therapy for promoting bone repair due to its effect to regulate macrophages response.Although natural antimicrobial peptides(AMPs)offer a good solution,the unknown toxicity,high cost and exogenetic immune response hamper their applications in clinic.In this work,we fabricated a nanowire-like composite material,named P@C,by combining chitosan and puerarin via solid-phase reaction,which can finely mimic the bio-functions of AMPs.Chitosan,serving as the bacteria membrane puncture agent,and puerarin,serving as the LPS target agent,synergistically destroy the bacterial membrane structure and inhibit its recovery,thus endowing P@C with good antibacterial property.In addition,P@C possesses good osteoimmunomodulation due to its ability of LPS elimination and macrophage differentiation modulation.The in vivo results show that P@C can inhibit the LPS induced bone destruction in the Escherichia coli infected rat.P@C exhibits superior bone regeneration in Escherichia coli infected rat due to the comprehensive functions of its superior antibacterial property,and its ability of LPS elimination and immunomodulation.P@C can well mimic the functions of AMPs,which provides a novel and effective method for treating the PJI in clinic.
基金support from the National Natural Science Foundation of China(U21A20100,81772363,32000938)Science and Technology Commission of Shanghai Municipality,China(20ZR1465000)+2 种基金High-end Entrepreneurial and Innovative Teams of Ningbo High-level Talents Project(2018A-09-C)Shenzhen Science and Technology Funding(JCYJ20210324120009026)S&T Innovation 2025 Major Special Program of Ningbo(2018B10040)are acknowledged.
文摘The adverse immune response mediated by macrophages is one of the main factors that are prone to lead poor osseointegration of polyetheretherketone(PEEK)implants in clinic.Hence,endowing PEEK with immunomodulatory ability to avoid the adverse immune response becomes a promising strategy to promote bone repair.In this work,sulfonation and hydrothermal treatment were used to fabricate a 3D porous surface on PEEK and hydroxyapatite(HA)composited PEEK.The HA composited PEEK with 3D porous surface inhibited macrophages polarizing to M1 phenotype and downregulated inducible nitric oxide synthase protein expression,which led to a nitric oxide concentration reduction in culture medium of mouse bone marrow mesenchymal stem cells(mBMSCs)under coculture condition.The decrease of nitric oxide concentration could help to increase bone formation-related OSX and ALP genes expressions and decrease bone resorption-related MMP-9 and MMP-13 genes expressions via cAMP–PKA–RUNX2 pathway in mBMSCs.In summary,the HA composited PEEK with 3D porous surface has the potential to promote osteogenesis of PEEK through immunomodulation,which provides a promising strategy to improve the bone repair ability of PEEK.
基金the National Natural Science Foundation of China(51831011 and 31570973)National Science Foundation for Distinguished Young Scholars of China(51525207)Science and Technology Commission of Shanghai Municipality(19JC1415500,17441904000,and 18YF1426900)are acknowledged.
文摘Ds-block elements have been gaining increasing attention in the field of biomaterials modification,owing to their excellent biological properties,such as antibiosis,osteogenesis,etc.However,their function mechanisms are not well understood and conflicting conclusions were drawn by previous studies on this issue,which are mainly resulted from the inconsistent experimental conditions.In this work,three most widely used ds-block elements,copper,zinc,and silver were introduced on titanium substrate by plasma immersion ion implantation method to investigate the rule of ds-block elements in the immune responses.Results showed that the implanted samples could decrease the inflammatory responses compared with Ti sample.The trend of anti-inflammatory effects of macrophages on samples was in correlation with cellular ROS levels,which was induced by the implanted biomaterials and positively correlated with the number of valence electrons of ds-block elements.The co-culture experiments of macrophages and bone marrow mesenchymal stem cells showed that these two kinds of cells could enhance the anti-inflammation and osteogenesis of samples by the paracrine manner of PGE2.In general,in their steady states on titanium substrate(Cu2+,Zn2+,Ag),the ds-block elements with more valence electrons exhibit better anti-inflammatory and osteogenic effects.Moreover,molecular biology experiments indicate that the PGE2-related signaling pathway may contribute to the desired immunoregulation and osteoinduction capability of ds-block elements.These findings suggest a correlation between the number of valence electrons of ds-block elements and the relevant biological responses,which provides new insight into the selection of implanted ions and surface design of biomaterials.
基金the National Natural Science Foundation of China(32371397,32000938,U21A20100)The Fundamental Research Funds for The Central Universities(YG2023ZD29)+2 种基金Shenzhen Science and Technology Funding(JCYJ20210324120009026)Laboratory Open Fund of Key Technology and Materials in Minimally Invasive Spine Surgery(2024JZWC-ZDB03,2024JZWC-YBA04)Talent project of Shanghai Tongren Hospital(TRKYRC-xx02)are acknowledged.
文摘Infection and poor tissue repair are the key causes of percutaneous implantation failure. However, there is a lackof effective strategies to cope with due to its high requirements of sterilization, soft tissue healing, andosseointegration. In this work, L-arginine (L-Arg) was loaded onto a sulfonated polyetheretherketone (PEEK)surface to solve this issue. Under the infection condition, nitric oxide (NO) and reactive oxygen species (ROS) areproduced through catalyzing L-Arg by inducible nitric oxide synthase (iNOS) and thus play a role in bacteriasterilization. Under the tissue repair condition, L-Arg is catalyzed to ornithine by Arginase-1 (Arg-1), whichpromotes the proliferation and collagen secretion of L929 and rBMSCs. Notably, L-Arg loading samples couldpolarize macrophages to M1 and M2 in infection and tissue repair conditions, respectively. The results in vivoshow that the L-Arg loading samples could enhance infected soft tissue sealing and bone regeneration. Insummary, L-Arg loading sulfonated PEEK could polarize macrophage through metabolic reprogramming,providing multi-functions of antibacterial abilities, soft tissue repair, and bone regeneration, which gives a newidea to design percutaneous implantation materials.
基金This study was supported by grants from the National Natural Science Foundation of China(81403029)the Natural Science Foundation of Shanghai(19ZR1449100)+2 种基金the Science and Technology Commission of Shanghai Municipality(19JC1415500)the Science and Technology Commission of Shanghai Municipality(18410760600)the International Partnership Program of Chinese Academy of Sciences(Grant No.GJHZ1850).
文摘Polyetheretherketone(PEEK)has been used as an implant material because it has similar mechani-cal properties to natural bone.However,inferior osseointegration and bioinertness hamper the clinical application of PEEK.In this study,the surfaces of sulfonated three-dimensional(3D)PEEK porous structures were loaded with different concentrations of strontium ranelate,a compound commonly used in the treatment or prevention of osteoporosis by promoting bone formation and inhibiting bone resorption.Field-emission scanning electron microscopy was used to characterize the topography of the structures,elemental carbon,oxygen and strontium contents were mea-sured by X-ray photoelectron spectroscopy,and surface zeta potentials and water-contact angle were also measured.The results indicated that strontium ranelate was successfully loaded onto the 3D porous structures.In vitro cellular results showed that strontium ranelate-treated sulfonated PEEK(SP-SR)strengthened the adhesion of MC3T3-E1 cells.The activity of alkaline phosphatase,collagen secretion and extracellular matrix mineralization deposition of MC3T3-E1 cells were also improved on the surface of SP-SR.These results indicate that SP-SR could serve a new implant candidate for surgical treatment.
