背景:溶质载体家族1成员5(solute carrier family 1 member 5,SLC1A5)在多种疾病中发挥了潜在作用,但确切作用机制尚不清楚。构建稳定的SLC1A5过表达和敲低细胞模型可为深入研究SLC1A5在疾病中的确切作用机制以及发现潜在治疗靶点提供...背景:溶质载体家族1成员5(solute carrier family 1 member 5,SLC1A5)在多种疾病中发挥了潜在作用,但确切作用机制尚不清楚。构建稳定的SLC1A5过表达和敲低细胞模型可为深入研究SLC1A5在疾病中的确切作用机制以及发现潜在治疗靶点提供有力的实验工具。目的:构建小鼠SLC1A5过表达和敲低的慢病毒载体,以建立稳定转染的RAW264.7细胞株,为深入探讨SLC1A5在炎症中的作用提供实验基础。方法:根据SLC1A5基因序列设计合成引物并使用聚合酶链反应扩增该基因片段。将目的基因定向接入经Age I/Nhe I酶切的载体质粒GV492中构建重组慢病毒质粒,对阳性克隆进一步筛选后测序比对结果;pHelper1.0质粒载体、pHelper2.0质粒载体、目的质粒载体与293T细胞共同培养并转染,获得慢病毒原液进行包装和滴度测定;在此基础上,通过体外培养RAW264.7细胞,确定嘌呤霉素工作质量浓度;不同滴度的慢病毒分别与RAW264.7细胞共同培养,根据荧光强度确定转染效率;用嘌呤霉素挑选出稳定转染细胞,实时荧光定量聚合酶链反应和蛋白免疫印迹方法检测稳定转染细胞株的SLC1A5基因和蛋白表达水平。结果与结论:(1)测序序列与目的序列一致提示重组慢病毒载体构建成功;(2)过表达SLC1A5慢病毒的滴度为1×10~9 TU/mL,敲低SLC1A5慢病毒的滴度为3×10~9 TU/mL;(3)确定RAW264.7细胞嘌呤霉素工作质量浓度为3μg/mL;(4)过表达/敲低SLC1A5慢病毒转染RAW264.7细胞的最佳条件皆为HiTransG P转染增强液且感染复数值等于50;(5)过表达SLC1A5稳转细胞株中SLC1A5基因和蛋白的表达量明显上调,而敲低SLC1A5稳转细胞株中SLC1A5基因和蛋白的表达量显著下调。结果表明,成功构建了小鼠SLC1A5过表达和敲低的慢病毒载体并获得稳定转染的RAW264.7细胞株。展开更多
目的探讨原发性喉癌患者癌组织中微小核糖核酸-425-5p(miR-425-5p)/跨膜蛋白受体Patched1(PTCH1)轴分子与临床病理参数及预后的关系。方法前瞻性选取2018年7月至2021年6月新乡医学院第一附属医院收治的108例原发性喉癌患者作为研究对象...目的探讨原发性喉癌患者癌组织中微小核糖核酸-425-5p(miR-425-5p)/跨膜蛋白受体Patched1(PTCH1)轴分子与临床病理参数及预后的关系。方法前瞻性选取2018年7月至2021年6月新乡医学院第一附属医院收治的108例原发性喉癌患者作为研究对象。比较癌组织、癌旁组织及不同病理特征癌组织miR-425-5p、PTCH1 m RNA相对表达量,采用Spearman法分析miR-425-5p、PTCH1与临床病理特征的相关性,随访3年,统计所有患者的3年生存率,比较生存与死亡患者癌组织中的miR-425-5p、而PTCH1 m RNA相对表达量,并利用受试者工作特征(ROC)曲线获取miR-425-5p、PTCH1最佳截断值,采用KM曲线分析miR-425-5p、PTCH1与预后的关系。结果原发性喉癌患者癌组织中的miR-425-5p相对表达量为1.81±0.48,明显高于癌旁组织的1.08±0.23,PTCH1 m RNA相对表达量为1.21±0.36,明显低于癌旁组织的1.63±0.41,差异均有统计学意义(P<0.05);Ⅲ~Ⅳ期、淋巴结转移、低分化癌组织中的miR-425-5p相对表达量分别为1.97±0.46、2.09±0.42、2.14±0.46,明显高于Ⅰ~Ⅱ期、无淋巴结转移、中高分化癌组织的1.54±0.41、1.66±0.39、1.60±0.40,PTCH1 m RNA相对表达量分别为1.09±0.21、1.04±0.24、1.01±0.20,明显低于Ⅰ~Ⅱ期、无淋巴结转移、中高分化癌组织的1.42±0.25、1.30±0.27、1.34±0.23,差异均有统计学意义(P<0.05);Spearman法分析结果显示,miR-425-5p与临床分期、淋巴结转移呈正相关(r=0.663、0.702,P<0.05),与分化程度呈负相关(r=-0.681,P<0.05),PTCH1与临床分期、淋巴结转移呈负相关(r=-0.652、-0.711,P<0.05),与分化程度呈正相关(r=0.694,P<0.05);死亡患者癌组织中的miR-425-5p相对表达量为2.23±0.46,明显高于生存患者癌组织的1.67±0.38,而PTCH1 m RNA相对表达量为0.96±0.21,明显低于生存患者癌组织的1.30±0.34,差异均有统计学意义(P<0.05);ROC分析结果显示,miR-425-5p、PTCH1预测死亡的曲线下面积(AUC)分别为0.815(95%CI:0.727~0.884)、0.792(95%CI:0.702~0.865),最佳截断值分别为2.01、1.09;KM分析结果显示,miR-425-5p高表达、PTCH1低表达患者3年生存率均低于miR-425-5p低表达、PTCH1高表达患者,差异均有统计学意义(P<0.05)。结论miR-425-5p在原发性喉癌癌组织中表达上调,PTCH1表达下调,联合检测对预后具有一定预测价值,可作为临床评估病情、预测预后的辅助指标,以指导临床工作。展开更多
目的:荟萃分析方法评价5-羟色胺(5-HT)1A受体部分激动剂治疗功能性消化不良(FD)的临床疗效及安全性。方法:计算机检索PubMed、Web of Science、CNKI等数据库建库起公开发表的5-HT1A受体部分激动剂治疗FD的随机对照研究(RCT)。RevMan 5....目的:荟萃分析方法评价5-羟色胺(5-HT)1A受体部分激动剂治疗功能性消化不良(FD)的临床疗效及安全性。方法:计算机检索PubMed、Web of Science、CNKI等数据库建库起公开发表的5-HT1A受体部分激动剂治疗FD的随机对照研究(RCT)。RevMan 5.4软件对纳入的资料荟萃分析,并进行亚组分析、分层分析,评价5-HT1A受体部分激动剂对FD的疗效及安全性。结果:纳入19项RCT,共计1575例患者(治疗组801例,对照组774例)。荟萃分析显示:治疗组总有效率高于对照组(OR=4.18,95%CI:3.05~5.73,P<0.00001),而消化道症状评分(SMD=-1.30,95%CI:-1.95~-0.64,P=0.0001)、焦虑状态评分(SMD=-1.22,95%CI:-1.79~-0.65,P<0.0001)和抑郁状态评分(SMD=-1.52,95%CI:-2.41~-0.63,P=0.0008)均低于对照组,嗜睡(OR=4.78,95%CI:1.80~12.70,P<0.05)、口干(OR=3.07,95%CI:1.31~7.19,P<0.05)发生率均高于对照组。结论:与常规或安慰剂治疗相比,联合应用5-HT1A受体部分激动剂能提高总体疗效,但嗜睡及口干发生率较高。展开更多
In patients with Alzheimer’s disease,gamma-glutamyl transferase 5(GGT5)expression has been observed to be downregulated in cerebrovascular endothelial cells.However,the functional role of GGT5 in the development of A...In patients with Alzheimer’s disease,gamma-glutamyl transferase 5(GGT5)expression has been observed to be downregulated in cerebrovascular endothelial cells.However,the functional role of GGT5 in the development of Alzheimer’s disease remains unclear.This study aimed to explore the effect of GGT5 on cognitive function and brain pathology in an APP/PS1 mouse model of Alzheimer’s disease,as well as the underlying mechanism.We observed a significant reduction in GGT5 expression in two in vitro models of Alzheimer’s disease(Aβ_(1-42)-treated hCMEC/D3 and bEnd.3 cells),as well as in the APP/PS1 mouse model.Additionally,injection of APP/PS1 mice with an adeno-associated virus encoding GGT5 enhanced hippocampal synaptic plasticity and mitigated cognitive deficits.Interestingly,increasing GGT5 expression in cerebrovascular endothelial cells reduced levels of both soluble and insoluble amyloid-βin the brains of APP/PS1 mice.This effect may be attributable to inhibition of the expression ofβ-site APP cleaving enzyme 1,which is mediated by nuclear factor-kappa B.Our findings demonstrate that GGT5 expression in cerebrovascular endothelial cells is inversely associated with Alzheimer’s disease pathogenesis,and that GGT5 upregulation mitigates cognitive deficits in APP/PS1 mice.These findings suggest that GGT5 expression in cerebrovascular endothelial cells is a potential therapeutic target and biomarker for Alzheimer’s disease.展开更多
The essential photoprotective role of proton gradient regulation 5(PGR5)-dependent cyclic electron flow(CEF)has been reported in Arabidopsis,rice,and algae.However,its functional assessment has not been performed in t...The essential photoprotective role of proton gradient regulation 5(PGR5)-dependent cyclic electron flow(CEF)has been reported in Arabidopsis,rice,and algae.However,its functional assessment has not been performed in tomato yet.In this study,we focused on elucidate the function of SlPGR5 and SlPGR5-like photosynthetic phenotype 1(PGRL1)in tomato.We performed RNA interference and found that SlPGR5/SlPGRL1-suppressed transformants exhibited extremely low CO_(2)assimilation capacity,their photosystem I(PSI)and PSII were severely photoinhibited and chloroplasts were obviously damaged.The SlPGR5/SlPGRL1-suppressed plants almost completely inhibited CEF and Y(ND),and PSII photoinhibition may be directly related to the inability to produce sufficient proton motive force to induce NPQ.The transgenic plants overexpressing SlPGR5 and SlPGRL1 driven by 35S promoter capable alleviate photoinhibition of plants under low night temperature.The transcriptomic and proteomic analyses suggested that the nuclear gene transcription and turnover of chloroplast proteins,including the plastoglobule-related proteins,were closely related to SlPGR5/SlPGRL1 pathway dependent CEF.The bridge relationship between CEF and chloroplast quality maintenance was a novel report to our knowledge.In conclusion,these results revealed the regulatory mechanism of the SlPGR5/SlPGRL1 pathway in photoprotection and maintenance of chloroplast function in tomato,which is crucial for reduce yield loss,especially under adverse environmental conditions.展开更多
文摘目的探讨原发性喉癌患者癌组织中微小核糖核酸-425-5p(miR-425-5p)/跨膜蛋白受体Patched1(PTCH1)轴分子与临床病理参数及预后的关系。方法前瞻性选取2018年7月至2021年6月新乡医学院第一附属医院收治的108例原发性喉癌患者作为研究对象。比较癌组织、癌旁组织及不同病理特征癌组织miR-425-5p、PTCH1 m RNA相对表达量,采用Spearman法分析miR-425-5p、PTCH1与临床病理特征的相关性,随访3年,统计所有患者的3年生存率,比较生存与死亡患者癌组织中的miR-425-5p、而PTCH1 m RNA相对表达量,并利用受试者工作特征(ROC)曲线获取miR-425-5p、PTCH1最佳截断值,采用KM曲线分析miR-425-5p、PTCH1与预后的关系。结果原发性喉癌患者癌组织中的miR-425-5p相对表达量为1.81±0.48,明显高于癌旁组织的1.08±0.23,PTCH1 m RNA相对表达量为1.21±0.36,明显低于癌旁组织的1.63±0.41,差异均有统计学意义(P<0.05);Ⅲ~Ⅳ期、淋巴结转移、低分化癌组织中的miR-425-5p相对表达量分别为1.97±0.46、2.09±0.42、2.14±0.46,明显高于Ⅰ~Ⅱ期、无淋巴结转移、中高分化癌组织的1.54±0.41、1.66±0.39、1.60±0.40,PTCH1 m RNA相对表达量分别为1.09±0.21、1.04±0.24、1.01±0.20,明显低于Ⅰ~Ⅱ期、无淋巴结转移、中高分化癌组织的1.42±0.25、1.30±0.27、1.34±0.23,差异均有统计学意义(P<0.05);Spearman法分析结果显示,miR-425-5p与临床分期、淋巴结转移呈正相关(r=0.663、0.702,P<0.05),与分化程度呈负相关(r=-0.681,P<0.05),PTCH1与临床分期、淋巴结转移呈负相关(r=-0.652、-0.711,P<0.05),与分化程度呈正相关(r=0.694,P<0.05);死亡患者癌组织中的miR-425-5p相对表达量为2.23±0.46,明显高于生存患者癌组织的1.67±0.38,而PTCH1 m RNA相对表达量为0.96±0.21,明显低于生存患者癌组织的1.30±0.34,差异均有统计学意义(P<0.05);ROC分析结果显示,miR-425-5p、PTCH1预测死亡的曲线下面积(AUC)分别为0.815(95%CI:0.727~0.884)、0.792(95%CI:0.702~0.865),最佳截断值分别为2.01、1.09;KM分析结果显示,miR-425-5p高表达、PTCH1低表达患者3年生存率均低于miR-425-5p低表达、PTCH1高表达患者,差异均有统计学意义(P<0.05)。结论miR-425-5p在原发性喉癌癌组织中表达上调,PTCH1表达下调,联合检测对预后具有一定预测价值,可作为临床评估病情、预测预后的辅助指标,以指导临床工作。
文摘目的:荟萃分析方法评价5-羟色胺(5-HT)1A受体部分激动剂治疗功能性消化不良(FD)的临床疗效及安全性。方法:计算机检索PubMed、Web of Science、CNKI等数据库建库起公开发表的5-HT1A受体部分激动剂治疗FD的随机对照研究(RCT)。RevMan 5.4软件对纳入的资料荟萃分析,并进行亚组分析、分层分析,评价5-HT1A受体部分激动剂对FD的疗效及安全性。结果:纳入19项RCT,共计1575例患者(治疗组801例,对照组774例)。荟萃分析显示:治疗组总有效率高于对照组(OR=4.18,95%CI:3.05~5.73,P<0.00001),而消化道症状评分(SMD=-1.30,95%CI:-1.95~-0.64,P=0.0001)、焦虑状态评分(SMD=-1.22,95%CI:-1.79~-0.65,P<0.0001)和抑郁状态评分(SMD=-1.52,95%CI:-2.41~-0.63,P=0.0008)均低于对照组,嗜睡(OR=4.78,95%CI:1.80~12.70,P<0.05)、口干(OR=3.07,95%CI:1.31~7.19,P<0.05)发生率均高于对照组。结论:与常规或安慰剂治疗相比,联合应用5-HT1A受体部分激动剂能提高总体疗效,但嗜睡及口干发生率较高。
基金supported by STI2030-Major Projects,No.2021ZD 0201801(to JG)Shanxi Province Basic Research Program,No.20210302123429(to QS).
文摘In patients with Alzheimer’s disease,gamma-glutamyl transferase 5(GGT5)expression has been observed to be downregulated in cerebrovascular endothelial cells.However,the functional role of GGT5 in the development of Alzheimer’s disease remains unclear.This study aimed to explore the effect of GGT5 on cognitive function and brain pathology in an APP/PS1 mouse model of Alzheimer’s disease,as well as the underlying mechanism.We observed a significant reduction in GGT5 expression in two in vitro models of Alzheimer’s disease(Aβ_(1-42)-treated hCMEC/D3 and bEnd.3 cells),as well as in the APP/PS1 mouse model.Additionally,injection of APP/PS1 mice with an adeno-associated virus encoding GGT5 enhanced hippocampal synaptic plasticity and mitigated cognitive deficits.Interestingly,increasing GGT5 expression in cerebrovascular endothelial cells reduced levels of both soluble and insoluble amyloid-βin the brains of APP/PS1 mice.This effect may be attributable to inhibition of the expression ofβ-site APP cleaving enzyme 1,which is mediated by nuclear factor-kappa B.Our findings demonstrate that GGT5 expression in cerebrovascular endothelial cells is inversely associated with Alzheimer’s disease pathogenesis,and that GGT5 upregulation mitigates cognitive deficits in APP/PS1 mice.These findings suggest that GGT5 expression in cerebrovascular endothelial cells is a potential therapeutic target and biomarker for Alzheimer’s disease.
基金supported by the National Natural Science Foundation of China(Grant Nos.32072651,31772356)China Agriculture Research System of MOF and MARA(Grant No.CARS23)+1 种基金Joint Fund for Innovation Enhancement of Liaoning Province(Grant No.2021-NLTS-11-01)Support Program for Young and middle-aged Scientific and technological Innovation Talents(Grant No.RC210293)。
文摘The essential photoprotective role of proton gradient regulation 5(PGR5)-dependent cyclic electron flow(CEF)has been reported in Arabidopsis,rice,and algae.However,its functional assessment has not been performed in tomato yet.In this study,we focused on elucidate the function of SlPGR5 and SlPGR5-like photosynthetic phenotype 1(PGRL1)in tomato.We performed RNA interference and found that SlPGR5/SlPGRL1-suppressed transformants exhibited extremely low CO_(2)assimilation capacity,their photosystem I(PSI)and PSII were severely photoinhibited and chloroplasts were obviously damaged.The SlPGR5/SlPGRL1-suppressed plants almost completely inhibited CEF and Y(ND),and PSII photoinhibition may be directly related to the inability to produce sufficient proton motive force to induce NPQ.The transgenic plants overexpressing SlPGR5 and SlPGRL1 driven by 35S promoter capable alleviate photoinhibition of plants under low night temperature.The transcriptomic and proteomic analyses suggested that the nuclear gene transcription and turnover of chloroplast proteins,including the plastoglobule-related proteins,were closely related to SlPGR5/SlPGRL1 pathway dependent CEF.The bridge relationship between CEF and chloroplast quality maintenance was a novel report to our knowledge.In conclusion,these results revealed the regulatory mechanism of the SlPGR5/SlPGRL1 pathway in photoprotection and maintenance of chloroplast function in tomato,which is crucial for reduce yield loss,especially under adverse environmental conditions.
