目的研究呼出气一氧化氮(FeNO)、血清免疫球蛋白E(IgE)和基质金属蛋白酶9(MMP9)的水平与儿童哮喘急性发作之间的关系,为儿童哮喘的预防及治疗提供依据。方法选取沈阳市妇婴医院于2020年11月至2022年11月收治的98例支气管哮喘急性发作期...目的研究呼出气一氧化氮(FeNO)、血清免疫球蛋白E(IgE)和基质金属蛋白酶9(MMP9)的水平与儿童哮喘急性发作之间的关系,为儿童哮喘的预防及治疗提供依据。方法选取沈阳市妇婴医院于2020年11月至2022年11月收治的98例支气管哮喘急性发作期儿童作为急性组,按照病情程度分成轻度组(n=32)、中度组(n=38)和重度组(n=28),按照2∶1的比例选出49例同期在门诊治疗的处于支气管哮喘缓解期的儿童作为缓解组,随机选取健康体检儿童49例作为健康对照组,分别对他们进行FeNO、MMP9和血清IgE及肺功能[用力肺活量(FVC)、1秒用力呼气量(FEV_(1))、FEV_(1)/FVC%、最大呼气流量(PEF)]检测。应用Pearson相关分析探讨哮喘急性发作期FeNO、MMP9及血清IgE和肺功能之间的联系,并对三者在支气管哮喘急性发作中的预测价值进行分析。结果急性组、缓解组和对照组的年龄、性别、体重指数和病程的比较,差异无统计学意义(P>0.05)。急性组FeNO、MMP9、血清IgE分别为(59.95±12.65)ppb、(4.87±1.44)pg/ml、(330.63±74.88)IU/ml,缓解组分别为(25.23±8.23)ppb、(1.21±0.02)pg/ml、(152.23±32.12)IU/ml,均高于对照组的(12.43±4.09)ppb、(0.53±0.24)pg/ml、(126.34±57.33)IU/ml,差异具有统计学意义(P<0.05)。急性期和缓解期FVC、FEV1、FEV1/FVC%、PEF均低于对照组,差异具有统计学意义(P<0.05)。支气管哮喘急性发作中度组FeNO、MMP9、血清IgE水平分别为(49.23±6.23)ppb、(1.21±0.02)pg/ml、(282.61±59.83)IU/ml,重度组分别为(67.43±10.09)ppb、(0.53±0.24)pg/ml、(356.49±70.82)IU/ml,均高于轻度组的(34.62±10.65)ppb、(4.87±1.44)pg/ml,(189.21±14.33)IU/ml,差异具有统计学意义(P<0.05)。在轻度组中FeNO、MMP9和血清IgE水平均较低,而在中度组中这些指标均较高,其中FVC、FEV_(1)、FEV_(1)/FVC%和PEF均较低,差异具有统计学意义(P<0.05)。FeNO以及MMP9与血清IgE水平呈正相关(P<0.05),FeNO、MMP9以及血清IgE水平与FVC、FEV_(1)、FEV_(1)/FVC%、PEF均呈负相关(P<0.05)。MMP9在支气管哮喘的诊断中表现出了显著的优势,当达到最大约登指数时,对应的截断值为1.17,曲线下面积(Area under curve,AUC)为0.83,敏感度和特异性也分别达到了90.13%和86.5%。结论支气管哮喘急性发作的儿童血清中的FeNO、MMP9以及血清IgE水平显著增高,随肺部功能恶化程度加重而上升,可能与支气管哮喘急性发作患儿肺功能损害程度有关。展开更多
Cotton is a pivotal economic crop for natural textile fibers that also serves as an important source of edible oil(Long et al.2023).Cottonseed oil contains approximately14%oleic acid and 59%linoleic acid.An increase i...Cotton is a pivotal economic crop for natural textile fibers that also serves as an important source of edible oil(Long et al.2023).Cottonseed oil contains approximately14%oleic acid and 59%linoleic acid.An increase in monounsaturated fatty acids,particularly oleic acid,enhances the oxidative stability and nutritional value of edible oil(Chen et al.2021).展开更多
Background Genome editing has been considered as powerful tool in agricultural fields.However,genome editing progress in cattle has not been fast as in other mammal species,for some disadvantages including long gestat...Background Genome editing has been considered as powerful tool in agricultural fields.However,genome editing progress in cattle has not been fast as in other mammal species,for some disadvantages including long gestational periods,single pregnancy,and high raising cost.Furthermore,technically demanding methods such as microinjection and somatic cell nuclear transfer(SCNT)are needed for gene editing in cattle.In this point of view,electroporation in embryos has been risen as an alternative.Results First,editing efficiency of our electroporation methods were tested for embryos.Presence of mutation on embryo was confirmed by T7E1 assay.With first combination,mutation rates for MSTN and PRNP were 57.6%±13.7%and 54.6%±13.5%,respectively.In case of MSTN/BLG,mutation rates were 83.9%±23.6%for MSTN,84.5%±18.0%for BLG.Afterwards,the double-KO embryos were transferred to surrogates and mutation rate was identified in resultant calves by targeted deep sequencing.Thirteen recipients were transferred for MSTN/PRNP,4 calves were delivered,and one calf underwent an induction for double KO.Ten surrogates were given double-KO embryos for MSTN/BLG,and four of the six calves that were born had mutations in both genes.Conclusions These data demonstrated that production of genome edited cattle via electroporation of RNP could be effectively applied.Finally,MSTN and PRNP from beef cattle and MSTN and BLG from dairy cattle have been born and they will be valuable resources for future precision breeding.展开更多
Noncoding RNAs instruct the Cas9 nuclease to site speifillyl cleave DNA in the CRISPR/Cas9 system.Despite the high incidence of hepatocellular carcinoma(HCC),the patient's outcome is poor.As a result of the emerge...Noncoding RNAs instruct the Cas9 nuclease to site speifillyl cleave DNA in the CRISPR/Cas9 system.Despite the high incidence of hepatocellular carcinoma(HCC),the patient's outcome is poor.As a result of the emergence of therapeutic resistance in HCC patients,dlinicians have faced difficulties in treating such tumor.In addition,CRISPR/Cas9 screens were used to identify genes that improve the dlinical response of HCC patients.It is the objective of this article to summarize the current understanding of the use of the CRISPR/Cas9 system for the treatment of cancer,with a particular emphasis on HCC as part of the current state of knowledge.Thus,in order to locate recent developments in oncology research,we examined both the Scopus database and the PubMed database.The ability to selectively interfere with gene expression in combinatorial CRISPR/Cas9 screening can lead to the discovery of new effective HCC treatment regimens by combining clinically approved drugs.Drug resistance can be overcome with the help of the CRISPR/Cas9 system.HCC signature genes and resistance to treatment have been uncovered by genome-scale CRISPR activation screening although this method is not without limitations.It has been extensively examined whether CRISPR can be used as a tool for disease research and gene therapy.CRISPR and its applications to tumor research,particularly in HCC,are examined in this study through a review of the literature.展开更多
Changes in lipid metabolism have been implicated in protection against infectious diseases. In the first experiment of this study, we measured clinical lipid parameters in a murine model where the unmethylated cytidin...Changes in lipid metabolism have been implicated in protection against infectious diseases. In the first experiment of this study, we measured clinical lipid parameters in a murine model where the unmethylated cytidine phosphate guanosine (CpG) oligodinucleotide (ODN1826), a Toll-like receptor 9 (TLR9) agonist was administered in combination with D-galactosamine (GalN) that caused relatively liver-specific inflammation and toxicity. In the control mice group injected with phosphate-buffered saline (PBS) (acute psychological stress model associated with blood sampling), the serum triglyceride (TG) levels showed a rapid decrease followed by a rebound at 24 h as we have recently reported. However, such a TG rebound was impaired in the CpG/GalN- and solely CpG-treated groups of mice despite an absence of liver injury based on serum alanine aminotransferase levels in the latter group. Thus, the stress-associated serum TG rebound was abrogated by the injection of a sub-hepatotoxic CpG dose. In the second experiment, we simply measured the hepatic CD36 and SACRB1 (the gene for scavenger receptor B1 (SR-B1)) transcripts after the i.p. administration of PBS, CpG or CpG/GalN. There was a remarkable elevation of hepatic CD36 transcript expression in both the CpG- and CpG/GalN-treated mice at 8 h post-CpG injection whereas the increase in the PBS-treated mice was slower than the former two groups, suggesting that hepatic CD36 transcript expression is more pronounced in the combined stress models than under psychological stress alone. The individual mice data showed that the increase in CD36 expression was accompanied by a reduction in SCARB1 mRNA, showing reciprocal regulation between these two genes. Together with our previously reported findings, these data suggest that, in a murine model combining psychological stress with TLR-triggered hepatic inflammation, the psychological stress facilitates liver uptake of plasma TG (and its components fatty acids), but the subsequent re-esterification and/or release of TG-rich lipoproteins from the liver is impaired due to the concomitant TLR-signaling. We hypothesize that lipid metabolism during acute stress shifts toward an elevated hepatic uptake of lipids due to concomitant TLR signaling, facilitating the clearance of bacterial lipids by the liver.展开更多
H9N2 virus has been widely distributed in wild birds and poultry around the world since its first emergence in the United States of America in 1966(Gu et al.2017;Carnaccini and Perez 2020).The virus appeared in chicke...H9N2 virus has been widely distributed in wild birds and poultry around the world since its first emergence in the United States of America in 1966(Gu et al.2017;Carnaccini and Perez 2020).The virus appeared in chickens in China in the early 1990s,and over the last two decades has gradually become the dominant epidemic subtype(Sun and Liu 2015;Bi et al.2020).Although H9N2 virus infection alone cannot cause severe disease or death in poultry,H9N2 virus-infected birds experience a degree of egg production drop and can be easily infected by other pathogens,thus causing economic losses for poultry industry.展开更多
The development and maturation of the CRISPR/Cas genome editing system provides a valuable tool for plant functional genomics and genetic improvement.Currently available genome-editing tools have a limited number of t...The development and maturation of the CRISPR/Cas genome editing system provides a valuable tool for plant functional genomics and genetic improvement.Currently available genome-editing tools have a limited number of targets,restricting their application in genetic research.In this study,we developed a novel CRISPR/Cas9 plant ultra-multiplex genome editing system consisting of two template vectors,eight donor vectors,four destination vectors,and one primer-design software package.By combining the advantages of Golden Gate cloning to assemble multiple repetitive fragments and Gateway recombination to assemble large fragments and by changing the structure of the amplicons used to assemble sg RNA expression cassettes,the plant ultra-multiplex genome editing system can assemble a single binary vector targeting more than 40 genomic loci.A rice knockout vector containing 49 sg RNA expression cassettes was assembled and a high co-editing efficiency was observed.This plant ultra-multiplex genome editing system advances synthetic biology and plant genetic engineering.展开更多
Soybean is a broadly popular and extensively cultivated crop,however,many high-yield and high-quality varieties require specific growth conditions,restricting their widespread adoption.The appropriate light conditions...Soybean is a broadly popular and extensively cultivated crop,however,many high-yield and high-quality varieties require specific growth conditions,restricting their widespread adoption.The appropriate light conditions and photoperiod must be attained for these varieties to thrive in new environments.In this study,we employed CRISPR/Cas9 to design two sgRNAs aimed at knocking out the maturity-related gene E4 in a major American soybean variety called''Jack'',which belongs to maturity group MGII.E4 gene is primarily involved in the photoperiodic flowering and maturity in soybean,making it an ideal candidate for genetic manipulation.We successfully obtained 1 homozygous E4-SG1 mutant type with 1-bp insertion,and 4 homozygous E4-SG2 mutants type with 2-bp deletion,7-bp deletion,61-bp deletion,and 1-bp insertion,respectively.The homozygous e4 mutant plants contained early termination codons devoid of transgenic elements.Additionally,no potential offtarget sites of the E4 gene were detected.A comparative analysis revealed that,unlike the wild-type,the maturity time of homozygous e4 mutants was early under both short-day and long-day conditions.These mutants offer novel germplasm resources that may be used to modify the photoperiod sensitivity and maturity of soybean,enhancing its adaptability to high-latitude regions.展开更多
文摘目的研究呼出气一氧化氮(FeNO)、血清免疫球蛋白E(IgE)和基质金属蛋白酶9(MMP9)的水平与儿童哮喘急性发作之间的关系,为儿童哮喘的预防及治疗提供依据。方法选取沈阳市妇婴医院于2020年11月至2022年11月收治的98例支气管哮喘急性发作期儿童作为急性组,按照病情程度分成轻度组(n=32)、中度组(n=38)和重度组(n=28),按照2∶1的比例选出49例同期在门诊治疗的处于支气管哮喘缓解期的儿童作为缓解组,随机选取健康体检儿童49例作为健康对照组,分别对他们进行FeNO、MMP9和血清IgE及肺功能[用力肺活量(FVC)、1秒用力呼气量(FEV_(1))、FEV_(1)/FVC%、最大呼气流量(PEF)]检测。应用Pearson相关分析探讨哮喘急性发作期FeNO、MMP9及血清IgE和肺功能之间的联系,并对三者在支气管哮喘急性发作中的预测价值进行分析。结果急性组、缓解组和对照组的年龄、性别、体重指数和病程的比较,差异无统计学意义(P>0.05)。急性组FeNO、MMP9、血清IgE分别为(59.95±12.65)ppb、(4.87±1.44)pg/ml、(330.63±74.88)IU/ml,缓解组分别为(25.23±8.23)ppb、(1.21±0.02)pg/ml、(152.23±32.12)IU/ml,均高于对照组的(12.43±4.09)ppb、(0.53±0.24)pg/ml、(126.34±57.33)IU/ml,差异具有统计学意义(P<0.05)。急性期和缓解期FVC、FEV1、FEV1/FVC%、PEF均低于对照组,差异具有统计学意义(P<0.05)。支气管哮喘急性发作中度组FeNO、MMP9、血清IgE水平分别为(49.23±6.23)ppb、(1.21±0.02)pg/ml、(282.61±59.83)IU/ml,重度组分别为(67.43±10.09)ppb、(0.53±0.24)pg/ml、(356.49±70.82)IU/ml,均高于轻度组的(34.62±10.65)ppb、(4.87±1.44)pg/ml,(189.21±14.33)IU/ml,差异具有统计学意义(P<0.05)。在轻度组中FeNO、MMP9和血清IgE水平均较低,而在中度组中这些指标均较高,其中FVC、FEV_(1)、FEV_(1)/FVC%和PEF均较低,差异具有统计学意义(P<0.05)。FeNO以及MMP9与血清IgE水平呈正相关(P<0.05),FeNO、MMP9以及血清IgE水平与FVC、FEV_(1)、FEV_(1)/FVC%、PEF均呈负相关(P<0.05)。MMP9在支气管哮喘的诊断中表现出了显著的优势,当达到最大约登指数时,对应的截断值为1.17,曲线下面积(Area under curve,AUC)为0.83,敏感度和特异性也分别达到了90.13%和86.5%。结论支气管哮喘急性发作的儿童血清中的FeNO、MMP9以及血清IgE水平显著增高,随肺部功能恶化程度加重而上升,可能与支气管哮喘急性发作患儿肺功能损害程度有关。
基金supported by the Science and Technology Innovation Talents in Universities of Henan Province,China(24HASTIT053)the National Natural Science Foundation of China(32172041)+1 种基金the Natural Science Foundation of Henan Province,China(232300421026)the Science and Technology Innovation 2030,China(2022ZD0402001-04)。
文摘Cotton is a pivotal economic crop for natural textile fibers that also serves as an important source of edible oil(Long et al.2023).Cottonseed oil contains approximately14%oleic acid and 59%linoleic acid.An increase in monounsaturated fatty acids,particularly oleic acid,enhances the oxidative stability and nutritional value of edible oil(Chen et al.2021).
基金financially supported by the National Research Foundation of Korea(NRF-2021R1A5A1033157 for SRC program:382 Comparative medicine Disease Research Center,NRF-2021R1F1A105195313)the Research Institute of Veterinary Science,the BK21 Four for Future Veterinary Medicine Leading Education and Research Center,and a Seoul National University(SNU)grant(#550e2020005)。
文摘Background Genome editing has been considered as powerful tool in agricultural fields.However,genome editing progress in cattle has not been fast as in other mammal species,for some disadvantages including long gestational periods,single pregnancy,and high raising cost.Furthermore,technically demanding methods such as microinjection and somatic cell nuclear transfer(SCNT)are needed for gene editing in cattle.In this point of view,electroporation in embryos has been risen as an alternative.Results First,editing efficiency of our electroporation methods were tested for embryos.Presence of mutation on embryo was confirmed by T7E1 assay.With first combination,mutation rates for MSTN and PRNP were 57.6%±13.7%and 54.6%±13.5%,respectively.In case of MSTN/BLG,mutation rates were 83.9%±23.6%for MSTN,84.5%±18.0%for BLG.Afterwards,the double-KO embryos were transferred to surrogates and mutation rate was identified in resultant calves by targeted deep sequencing.Thirteen recipients were transferred for MSTN/PRNP,4 calves were delivered,and one calf underwent an induction for double KO.Ten surrogates were given double-KO embryos for MSTN/BLG,and four of the six calves that were born had mutations in both genes.Conclusions These data demonstrated that production of genome edited cattle via electroporation of RNP could be effectively applied.Finally,MSTN and PRNP from beef cattle and MSTN and BLG from dairy cattle have been born and they will be valuable resources for future precision breeding.
文摘Noncoding RNAs instruct the Cas9 nuclease to site speifillyl cleave DNA in the CRISPR/Cas9 system.Despite the high incidence of hepatocellular carcinoma(HCC),the patient's outcome is poor.As a result of the emergence of therapeutic resistance in HCC patients,dlinicians have faced difficulties in treating such tumor.In addition,CRISPR/Cas9 screens were used to identify genes that improve the dlinical response of HCC patients.It is the objective of this article to summarize the current understanding of the use of the CRISPR/Cas9 system for the treatment of cancer,with a particular emphasis on HCC as part of the current state of knowledge.Thus,in order to locate recent developments in oncology research,we examined both the Scopus database and the PubMed database.The ability to selectively interfere with gene expression in combinatorial CRISPR/Cas9 screening can lead to the discovery of new effective HCC treatment regimens by combining clinically approved drugs.Drug resistance can be overcome with the help of the CRISPR/Cas9 system.HCC signature genes and resistance to treatment have been uncovered by genome-scale CRISPR activation screening although this method is not without limitations.It has been extensively examined whether CRISPR can be used as a tool for disease research and gene therapy.CRISPR and its applications to tumor research,particularly in HCC,are examined in this study through a review of the literature.
文摘Changes in lipid metabolism have been implicated in protection against infectious diseases. In the first experiment of this study, we measured clinical lipid parameters in a murine model where the unmethylated cytidine phosphate guanosine (CpG) oligodinucleotide (ODN1826), a Toll-like receptor 9 (TLR9) agonist was administered in combination with D-galactosamine (GalN) that caused relatively liver-specific inflammation and toxicity. In the control mice group injected with phosphate-buffered saline (PBS) (acute psychological stress model associated with blood sampling), the serum triglyceride (TG) levels showed a rapid decrease followed by a rebound at 24 h as we have recently reported. However, such a TG rebound was impaired in the CpG/GalN- and solely CpG-treated groups of mice despite an absence of liver injury based on serum alanine aminotransferase levels in the latter group. Thus, the stress-associated serum TG rebound was abrogated by the injection of a sub-hepatotoxic CpG dose. In the second experiment, we simply measured the hepatic CD36 and SACRB1 (the gene for scavenger receptor B1 (SR-B1)) transcripts after the i.p. administration of PBS, CpG or CpG/GalN. There was a remarkable elevation of hepatic CD36 transcript expression in both the CpG- and CpG/GalN-treated mice at 8 h post-CpG injection whereas the increase in the PBS-treated mice was slower than the former two groups, suggesting that hepatic CD36 transcript expression is more pronounced in the combined stress models than under psychological stress alone. The individual mice data showed that the increase in CD36 expression was accompanied by a reduction in SCARB1 mRNA, showing reciprocal regulation between these two genes. Together with our previously reported findings, these data suggest that, in a murine model combining psychological stress with TLR-triggered hepatic inflammation, the psychological stress facilitates liver uptake of plasma TG (and its components fatty acids), but the subsequent re-esterification and/or release of TG-rich lipoproteins from the liver is impaired due to the concomitant TLR-signaling. We hypothesize that lipid metabolism during acute stress shifts toward an elevated hepatic uptake of lipids due to concomitant TLR signaling, facilitating the clearance of bacterial lipids by the liver.
基金supported by the National Key Research and Development Program of China(2021YFD1800200 and 2021YFC2301700)the National Natural Science Foundation of China(32192451)+1 种基金the Innovation Program of the Chinese Academy of Agricultural Sciences(CAASCSLPDCP-202301)the earmarked fund for CARS41(CARS-41).
文摘H9N2 virus has been widely distributed in wild birds and poultry around the world since its first emergence in the United States of America in 1966(Gu et al.2017;Carnaccini and Perez 2020).The virus appeared in chickens in China in the early 1990s,and over the last two decades has gradually become the dominant epidemic subtype(Sun and Liu 2015;Bi et al.2020).Although H9N2 virus infection alone cannot cause severe disease or death in poultry,H9N2 virus-infected birds experience a degree of egg production drop and can be easily infected by other pathogens,thus causing economic losses for poultry industry.
基金supported by the National Natural Science Foundation of China(32001532 and 31860411)the National Key Research and Development Program of China,(2022YFF1000020)+1 种基金Hunan Seed Industry Innovation Project(2021NK1012)the Yunnan Tobacco Company Project(2020530000241009)。
文摘The development and maturation of the CRISPR/Cas genome editing system provides a valuable tool for plant functional genomics and genetic improvement.Currently available genome-editing tools have a limited number of targets,restricting their application in genetic research.In this study,we developed a novel CRISPR/Cas9 plant ultra-multiplex genome editing system consisting of two template vectors,eight donor vectors,four destination vectors,and one primer-design software package.By combining the advantages of Golden Gate cloning to assemble multiple repetitive fragments and Gateway recombination to assemble large fragments and by changing the structure of the amplicons used to assemble sg RNA expression cassettes,the plant ultra-multiplex genome editing system can assemble a single binary vector targeting more than 40 genomic loci.A rice knockout vector containing 49 sg RNA expression cassettes was assembled and a high co-editing efficiency was observed.This plant ultra-multiplex genome editing system advances synthetic biology and plant genetic engineering.
基金supported by grants from the National Key R&D Program of China (2023YFD1201300)CAAS Agricultural Science and Technology Innovation Project
文摘Soybean is a broadly popular and extensively cultivated crop,however,many high-yield and high-quality varieties require specific growth conditions,restricting their widespread adoption.The appropriate light conditions and photoperiod must be attained for these varieties to thrive in new environments.In this study,we employed CRISPR/Cas9 to design two sgRNAs aimed at knocking out the maturity-related gene E4 in a major American soybean variety called''Jack'',which belongs to maturity group MGII.E4 gene is primarily involved in the photoperiodic flowering and maturity in soybean,making it an ideal candidate for genetic manipulation.We successfully obtained 1 homozygous E4-SG1 mutant type with 1-bp insertion,and 4 homozygous E4-SG2 mutants type with 2-bp deletion,7-bp deletion,61-bp deletion,and 1-bp insertion,respectively.The homozygous e4 mutant plants contained early termination codons devoid of transgenic elements.Additionally,no potential offtarget sites of the E4 gene were detected.A comparative analysis revealed that,unlike the wild-type,the maturity time of homozygous e4 mutants was early under both short-day and long-day conditions.These mutants offer novel germplasm resources that may be used to modify the photoperiod sensitivity and maturity of soybean,enhancing its adaptability to high-latitude regions.
