目的:对忍冬不同器官,即花蕾、叶和茎中HQT(hydroxycinnamoyl CoA quinate hydroxycinnamoyl transfer-ase)基因的相对表达量进行研究,以揭示HQT基因与忍冬绿原酸生物合成之间的相关性。方法:利用半定量RT-PCR方法对忍冬不同器官中的内...目的:对忍冬不同器官,即花蕾、叶和茎中HQT(hydroxycinnamoyl CoA quinate hydroxycinnamoyl transfer-ase)基因的相对表达量进行研究,以揭示HQT基因与忍冬绿原酸生物合成之间的相关性。方法:利用半定量RT-PCR方法对忍冬不同器官中的内参Actin基因以及HQT基因进行测定,通过琼脂糖凝胶电泳对PCR结果进行分析。结果:忍冬不同器官中Actin基因条带亮度比较接近,而HQT基因条带亮度差异较大,花蕾中HQT基因条带最亮,叶中次之,而茎中HQT基因条带非常微弱。此结果与绿原酸在忍冬不同器官中的含量相一致。结论:忍冬HQT基因的表达与绿原酸的合成之间可能存在有必然的联系。展开更多
目的通过构建羟基桂皮酰辅酶A羟基桂皮酰转移酶(hydroxycinnamoyl CoA quinate hydrocycinnamoyl transferase,HQT)基因真核表达载体以及建立金银花的遗传转化系统,阐明金银花HQT基因在真核生物细胞中对绿原酸量的调控机制。方法应用通...目的通过构建羟基桂皮酰辅酶A羟基桂皮酰转移酶(hydroxycinnamoyl CoA quinate hydrocycinnamoyl transferase,HQT)基因真核表达载体以及建立金银花的遗传转化系统,阐明金银花HQT基因在真核生物细胞中对绿原酸量的调控机制。方法应用通路克隆系统技术构建HQT基因真核表达载体,利用根癌农杆菌介导法,构建含金银花HQT基因的工程菌,对转染成功的外植体进行愈伤诱导,进而进行半定量RT-PCR检测转基因愈伤组织中HQT基因的相对表达量以及利用HPLC检测其绿原酸的量,并对二者相关性进行分析。结果金银花愈伤组织中绿原酸的量随HQT基因表达量的升高而升高。结论金银花HQT基因在真核生物细胞内对绿原酸的生物合成具备调控作用。展开更多
羟基肉桂酰辅酶A奎尼羟基肉桂转移酶(hydroxycinnamoyl Co A quinate hydroxycinnamoyl transferase,HQT)是金银花绿原酸合成过程中的关键酶,本研究利用生物信息学的分析方法,对金银花HQT氨基酸序列与其它植物间相应片段进行同源比对和...羟基肉桂酰辅酶A奎尼羟基肉桂转移酶(hydroxycinnamoyl Co A quinate hydroxycinnamoyl transferase,HQT)是金银花绿原酸合成过程中的关键酶,本研究利用生物信息学的分析方法,对金银花HQT氨基酸序列与其它植物间相应片段进行同源比对和系统进化分析,并对该基因编码蛋白的理化性质,亚细胞定位、跨膜结构、二级结构和三级结构等方面进行了预测和分析。结果表明,LjHQT编码的蛋白含424个氨基酸,是一种亲水性蛋白,蛋白磷酸化位点有7个,其中丝氨酸位点6个,酪氨酸1个;目的蛋白中不存在信号肽,没有跨膜结构域,LjHQT可能定位于叶绿体或其它细胞器;其编码的氨基酸序列与中果咖啡的同源关系较近,具有较高的保守性与乌梅、葡萄的同源关系较远。二三级结构分析表明,LjHQT编码的蛋白主要由无规则卷曲和α螺旋组成,空间结构主要是螺旋和转角。本研究为LjHQT功能分析提供了重要研究基础。展开更多
Background:Rheumatoid arthritis(RA)is a worldwide public health problem.Intervention and prevention before the onset of rheumatic diseases is a new direction in current research.Objective:The aim of this study was to ...Background:Rheumatoid arthritis(RA)is a worldwide public health problem.Intervention and prevention before the onset of rheumatic diseases is a new direction in current research.Objective:The aim of this study was to evaluate the potential and feasibility of traditional Chinese medicine(TCM)in the prevention of RA.Methods:This was a single-armed prospective clinical trial.All participants were recruited from a single center in Guangdong,China.Adults who were tested positive for anti-cyclic citrullinated peptide antibody(anti-CCP)and/or rheumatoid factor(RF),had no synovitis and never been treated with disease-modifying anti-rheumatic drugs(DMARDs),were enrolled to take the Huayu-Qiangshen-Tongbi(HQT)decoction orally twice daily,200 mL each time for 24 weeks.Primary outcome was the proportion of patients who met 2010 ACR(American College of Rheumatology)/EULAR(European League Against Rheumatism)classification criteria of RA during observation.Secondary outcomes included levels of anti-CCP,RF,erythrocyte sedimentation rate(ESR),C-reactive protein(CRP),assessment of signs and symptoms,and radiographic progression by magnetic resonance imaging(MRI).Results:19 individuals were enrolled in the study,4 of which withdrew because of the epidemic of COVID-19.During the observation period,3 individuals(20%)developed RA and they had longer morning stiffness(P=0.009)and more obvious synovial enhancement in MRI(P=0.041)at baseline when compared with those who did not develop RA.After 24 weeks of intervention,there were improvements in 28-swollen joint count(SJC28)(P=0.046),Visual Analog Scale(VAS)(P=0.019),Patient’s Global Assessment(PtGA)(P=0.019)and Physician’s Global Assessment(PGA)(P=0.031),but no statistical significance was observed in the levels of anti-CCP,RF,ESR,CRP,morning stiffness,28-tender joint count(TJC28),Health Assessment Questionnaire(HAQ)and magnetic resonance imaging(MRI)analysis(P>0.05).Conclusion:The HQT formula is safe and could improve joint symptoms and signs in these at-risk individuals,but it remains to be investigated in futher study to see if it might potentially reduce the risk of developing RA.Besides,for individuals at high risk to develop RA,morning stiffness and synovial enhancement in MRI might be predictive factors and warning signs.展开更多
文摘目的:对忍冬不同器官,即花蕾、叶和茎中HQT(hydroxycinnamoyl CoA quinate hydroxycinnamoyl transfer-ase)基因的相对表达量进行研究,以揭示HQT基因与忍冬绿原酸生物合成之间的相关性。方法:利用半定量RT-PCR方法对忍冬不同器官中的内参Actin基因以及HQT基因进行测定,通过琼脂糖凝胶电泳对PCR结果进行分析。结果:忍冬不同器官中Actin基因条带亮度比较接近,而HQT基因条带亮度差异较大,花蕾中HQT基因条带最亮,叶中次之,而茎中HQT基因条带非常微弱。此结果与绿原酸在忍冬不同器官中的含量相一致。结论:忍冬HQT基因的表达与绿原酸的合成之间可能存在有必然的联系。
文摘目的通过构建羟基桂皮酰辅酶A羟基桂皮酰转移酶(hydroxycinnamoyl CoA quinate hydrocycinnamoyl transferase,HQT)基因真核表达载体以及建立金银花的遗传转化系统,阐明金银花HQT基因在真核生物细胞中对绿原酸量的调控机制。方法应用通路克隆系统技术构建HQT基因真核表达载体,利用根癌农杆菌介导法,构建含金银花HQT基因的工程菌,对转染成功的外植体进行愈伤诱导,进而进行半定量RT-PCR检测转基因愈伤组织中HQT基因的相对表达量以及利用HPLC检测其绿原酸的量,并对二者相关性进行分析。结果金银花愈伤组织中绿原酸的量随HQT基因表达量的升高而升高。结论金银花HQT基因在真核生物细胞内对绿原酸的生物合成具备调控作用。
文摘羟基肉桂酰辅酶A奎尼羟基肉桂转移酶(hydroxycinnamoyl Co A quinate hydroxycinnamoyl transferase,HQT)是金银花绿原酸合成过程中的关键酶,本研究利用生物信息学的分析方法,对金银花HQT氨基酸序列与其它植物间相应片段进行同源比对和系统进化分析,并对该基因编码蛋白的理化性质,亚细胞定位、跨膜结构、二级结构和三级结构等方面进行了预测和分析。结果表明,LjHQT编码的蛋白含424个氨基酸,是一种亲水性蛋白,蛋白磷酸化位点有7个,其中丝氨酸位点6个,酪氨酸1个;目的蛋白中不存在信号肽,没有跨膜结构域,LjHQT可能定位于叶绿体或其它细胞器;其编码的氨基酸序列与中果咖啡的同源关系较近,具有较高的保守性与乌梅、葡萄的同源关系较远。二三级结构分析表明,LjHQT编码的蛋白主要由无规则卷曲和α螺旋组成,空间结构主要是螺旋和转角。本研究为LjHQT功能分析提供了重要研究基础。
基金This study was supported by the National Natural Science Foun-dation of China(81804041)the special project of State Key Laboratory of Dampness Syndrome of Chinese Medicine(SZ2020ZZ17)+5 种基金the 2020 Guangdong Provincial Science and Technology Innovation Strategy Special Fund(The Guangdong-Hong Kong-Macao Joint Lab)(2020B1212030006)Natural Science Foundation of Guang-dong Province(2021A1515011477,2021A1515011593)grant from Guangzhou Basic Research Program(202102010256)the Key Research Project of Guangzhou University of Chinese Medicine(XK2019021)opening project of Guangdong Provincial Key Laboratory of Chinese Medicine for Prevention and Treatment of Refractory Chronic Diseases(2018)(2018B030322012,MB2020KF03),Innovation Team and Talents Cultivation Program of National Administration of Traditional Chinese Medicine(ZYYCXTD-C-202204)as well as grants from Guangdong Provincial Hospital of Chinese Medicine(MB2019ZZ07,YN10101906,YN2018ML08,YN2018ZD06)。
文摘Background:Rheumatoid arthritis(RA)is a worldwide public health problem.Intervention and prevention before the onset of rheumatic diseases is a new direction in current research.Objective:The aim of this study was to evaluate the potential and feasibility of traditional Chinese medicine(TCM)in the prevention of RA.Methods:This was a single-armed prospective clinical trial.All participants were recruited from a single center in Guangdong,China.Adults who were tested positive for anti-cyclic citrullinated peptide antibody(anti-CCP)and/or rheumatoid factor(RF),had no synovitis and never been treated with disease-modifying anti-rheumatic drugs(DMARDs),were enrolled to take the Huayu-Qiangshen-Tongbi(HQT)decoction orally twice daily,200 mL each time for 24 weeks.Primary outcome was the proportion of patients who met 2010 ACR(American College of Rheumatology)/EULAR(European League Against Rheumatism)classification criteria of RA during observation.Secondary outcomes included levels of anti-CCP,RF,erythrocyte sedimentation rate(ESR),C-reactive protein(CRP),assessment of signs and symptoms,and radiographic progression by magnetic resonance imaging(MRI).Results:19 individuals were enrolled in the study,4 of which withdrew because of the epidemic of COVID-19.During the observation period,3 individuals(20%)developed RA and they had longer morning stiffness(P=0.009)and more obvious synovial enhancement in MRI(P=0.041)at baseline when compared with those who did not develop RA.After 24 weeks of intervention,there were improvements in 28-swollen joint count(SJC28)(P=0.046),Visual Analog Scale(VAS)(P=0.019),Patient’s Global Assessment(PtGA)(P=0.019)and Physician’s Global Assessment(PGA)(P=0.031),but no statistical significance was observed in the levels of anti-CCP,RF,ESR,CRP,morning stiffness,28-tender joint count(TJC28),Health Assessment Questionnaire(HAQ)and magnetic resonance imaging(MRI)analysis(P>0.05).Conclusion:The HQT formula is safe and could improve joint symptoms and signs in these at-risk individuals,but it remains to be investigated in futher study to see if it might potentially reduce the risk of developing RA.Besides,for individuals at high risk to develop RA,morning stiffness and synovial enhancement in MRI might be predictive factors and warning signs.
