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miR-23b-3p调控肾间质成纤维细胞成骨样分化参与Randall斑形成的机制研究 被引量:1
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作者 雷波 邱明星 刘健男 《安徽医科大学学报》 CAS 北大核心 2023年第12期2064-2072,共9页
目的探究miR-23b-3p对肾间质成纤维(hRIFs)细胞成骨样分化和Randall斑形成的影响及其可能机制。方法采用qRT-PCR技术检测草酸钙(CaOx)结石患者Randall斑组织(RP)和接受肾切除术的肾肿瘤患者正常乳头状组织(nRP)的miR-23b-3p和肌细胞特... 目的探究miR-23b-3p对肾间质成纤维(hRIFs)细胞成骨样分化和Randall斑形成的影响及其可能机制。方法采用qRT-PCR技术检测草酸钙(CaOx)结石患者Randall斑组织(RP)和接受肾切除术的肾肿瘤患者正常乳头状组织(nRP)的miR-23b-3p和肌细胞特异性增强子因子2C(MEF2C)、成骨标志物人骨钙蛋白(OCN)、骨桥蛋白(OPN)、Runt相关转录因子2(Runx2)的mRNA表达水平;体外分离、培养hRIFs细胞,将miR-23b-3p过表达质粒pSi-miR-23b-3p及空载质粒pSi-NC和MEF2C慢病毒过表达质粒Lv-MEF2C及空载质粒Lv-NC转染至hRIFs细胞中,并诱导细胞成骨分化14 d;ELISA法检测细胞碱性磷酸酶(ALP)活性;茜素红染色观察各组细胞矿化结节形成情况;qRT-PCR检测细胞中miR-23b-3p和MEF2C、OCN、OPN、Runx2 mRNA表达水平;Western blot检测细胞中MEF2C蛋白表达水平;双荧光素酶报告基因实验验证miR-23b-3p与MEF2C的靶向关系。结果①与nRP组比较,RP组中miR-23b-3p低表达,而MEF2C、OCN、OPN和Runx2 mRNA高表达;②hRIFs成骨诱导14 d后,细胞中ALP活性升高,矿化结节形成能力增强,miR-23b-3p表达水平降低,而MEF2C、OCN、OPN和Runx2 mRNA表达水平及MEF2C蛋白表达水平升高;③miR-23b-3p过表达降低成骨诱导后hRIFs细胞中ALP活性,抑制细胞矿化结节形成能力,下调细胞中OCN、OPN、Runx2 mRNA表达水平;④MEF2C过表达可逆转miR-23b-3p过表达对hRIFs细胞成骨分化的抑制作用;⑤MEF2C是miR-23b-3p下游靶基因。结论miR-23b-3p在RP组织及hRIFs细胞成骨样分化过程中低表达,上调miR-23b-3p可抑制hRIFs细胞的成骨样分化,其作用机制可能与靶向沉默MEF2C有关。 展开更多
关键词 miR-23b-3p 人肾间质成纤维细胞 肌细胞增强因子2C 成骨样分化 randall 草酸钙结石
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Rbm8a regulates neurogenesis and reduces Alzheimer's disease-associated pathology in the dentate gyrus of 5×FAD mice
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作者 Chenlu Zhu Xiao Ren +2 位作者 Chen Liu Yawei Liu Yonggang Wang 《Neural Regeneration Research》 SCIE CAS CSCD 2024年第4期863-871,共9页
Alzheimer’s disease is a prevalent and debilitating neurodegenerative condition that profoundly affects a patient’s daily functioning with progressive cognitive decline,which can be partly attributed to impaired hip... Alzheimer’s disease is a prevalent and debilitating neurodegenerative condition that profoundly affects a patient’s daily functioning with progressive cognitive decline,which can be partly attributed to impaired hippocampal neurogenesis.Neurogenesis in the hippocampal dentate gyrus is likely to persist throughout life but declines with aging,especially in Alzheimer’s disease.Recent evidence indicated that RNA-binding protein 8A(Rbm8a)promotes the proliferation of neural progenitor cells,with lower expression levels observed in Alzheimer’s disease patients compared with healthy people.This study investigated the hypothesis that Rbm8a overexpression may enhance neurogenesis by promoting the proliferation of neural progenitor cells to improve memory impairment in Alzheimer’s disease.Therefore,Rbm8a overexpression was induced in the dentate gyrus of 5×FAD mice to validate this hypothesis.Elevated Rbm8a levels in the dentate gyrus triggered neurogenesis and abated pathological phenotypes(such as plaque formation,gliosis reaction,and dystrophic neurites),leading to ameliorated memory performance in 5×FAD mice.RNA sequencing data further substantiated these findings,showing the enrichment of differentially expressed genes involved in biological processes including neurogenesis,cell proliferation,and amyloid protein formation.In conclusion,overexpressing Rbm8a in the dentate gyrus of 5×FAD mouse brains improved cognitive function by ameliorating amyloid-beta-associated pathological phenotypes and enhancing neurogenesis. 展开更多
关键词 Adora2a Alzheimer’s disease AsTROCYTE cAMP signaling pathway dentate gyrus dystrophic neurites MICROGLIA NEUROGENEsIs plaquE Rbm8a
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基质Gla蛋白与骨形成蛋白2在草酸钙肾结石患者Randall钙斑中的表达 被引量:5
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作者 朱敏 曾锋 +2 位作者 崔雨 刘细峰 陈合群 《中南大学学报(医学版)》 CAS CSCD 北大核心 2017年第3期277-283,共7页
目的:对比基质Gla蛋白(matrix Gla protein,MGP)及骨形成蛋白2(bone morphogenetic protein 2,BMP-2)在草酸钙结石肾乳头Randall钙化斑组织中的表达,探讨其在草酸钙肾结石形成过程中的相关机制。方法:收集中南大学湘雅医院泌尿外科2015... 目的:对比基质Gla蛋白(matrix Gla protein,MGP)及骨形成蛋白2(bone morphogenetic protein 2,BMP-2)在草酸钙结石肾乳头Randall钙化斑组织中的表达,探讨其在草酸钙肾结石形成过程中的相关机制。方法:收集中南大学湘雅医院泌尿外科2015年4月至12月期间住院的30例草酸钙肾结石患者的肾乳头钙化斑组织标本作为实验组,10例同期患肾肿瘤行根治性肾切除术患者的肾乳头组织标本作为对照组,利用实时荧光定量聚合酶链式反应(real-time polymerase chain reaction,RT-PCR)和Western印迹检测MGP和BMP-2在肾乳头组织中的表达,并采用免疫组织化学技术观察MGP和BMP-2在肾乳头组织中的表达。结果:1)RT-PCR检测实验组和对照组MGP mRNA的表达量为0.760±0.804和1.365±0.348,差异有统计学意义(P<0.05);实验组和对照组BMP-2 mRNA表达量分别为2.500±0.725和1.485±0.870,差异有统计学意义(P<0.05)。2)Western印迹检测实验组和对照组的MGP蛋白表达量分别为0.130±0.424和0.202±0.704,差异无统计学意义(P>0.05);实验组和对照组BMP-2蛋白表达量为0.885±0.220和0.682±0.272,差异有统计学意义(P<0.05)。3)免疫组织化学结果显示实验组MGP的表达较对照组低,而实验组BMP-2表达较对照组高(均P<0.05)。结论:草酸钙肾结石患者的肾乳头组织内BMP-2表达增多,MGP表达减少,草酸钙肾结石的形成可能是一种与MGP和BMP-2相关的类成骨反应或异位钙化。 展开更多
关键词 基质GLA蛋白 骨形成蛋白2 草酸钙肾结石 randall钙化斑
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Randall斑泌尿系结石患者的代谢评估 被引量:5
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作者 张一帆 许长宝 +4 位作者 万优 王曦堃 周德志 冀明阳 王友铭 《天津医药》 CAS 北大核心 2019年第10期1072-1075,共4页
目的通过对Randall斑结石患者进行24h尿代谢分析,探讨Randall斑患者的结石危险因素及机制。方法收集2017年10月—2018年10月于我院接受住院治疗的泌尿系结石患者50例,其中有Randall斑结石患者(Randall斑组)30例,无Randall斑结石患者(无R... 目的通过对Randall斑结石患者进行24h尿代谢分析,探讨Randall斑患者的结石危险因素及机制。方法收集2017年10月—2018年10月于我院接受住院治疗的泌尿系结石患者50例,其中有Randall斑结石患者(Randall斑组)30例,无Randall斑结石患者(无Randall斑组)20例,另择我院同期健康体检者30例为对照组。收集患者24h尿液进行尿液成分分析,对于排出的结石采用红外光谱分析仪检测结石成分。结果Randall斑组中结石的大小、位置、所在侧和结石成分与无Randall斑组差异均无统计学意义。与对照组相比,无Randall斑组24h尿液中枸橼酸含量降低,草酸、磷酸含量升高(P<0.05),Randall斑组24h尿液中枸橼酸含量降低,草酸、磷酸、钠、钙含量升高(P<0.05)。与无Randall斑组相比,Randall斑组24h尿液中枸橼酸含量降低,草酸、磷酸、钠、钙含量升高(P<0.05)。结论Randall斑患者24h尿液中高钠、钙、草酸、磷酸和低枸橼酸对结石的发生发展有重要影响,应限制Randall斑的泌尿系结石患者草酸、钠、钙、磷酸等的摄入并酌情补充枸橼酸。 展开更多
关键词 randall 泌尿系结石 24h尿液 危险因素
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肾结石Randall斑块研究进展 被引量:4
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作者 李永超 李杨 《中南大学学报(医学版)》 CAS CSCD 北大核心 2020年第4期435-439,共5页
泌尿系统结石具有高发病率和复发率的特点,是泌尿外科的难题。Randall斑块学说是目前学者们普遍认可的肾结石形成机制,Randall斑块形成的理论包括血管钙化学说、成骨转化学说等,目前尚未形成统一的学说。长链非编码RNA作为非编码RNA中... 泌尿系统结石具有高发病率和复发率的特点,是泌尿外科的难题。Randall斑块学说是目前学者们普遍认可的肾结石形成机制,Randall斑块形成的理论包括血管钙化学说、成骨转化学说等,目前尚未形成统一的学说。长链非编码RNA作为非编码RNA中一个重要的种类,它的突变或失调与疾病的发生和进展关系密切。与非结石患者肾乳头组织相比,肾结石患者肾乳头钙斑处组织中差异表达的长链非编码RNA提示其参与Randall斑块的形成。弹性纤维性假黄瘤是一种罕见的常染色体隐性遗传性疾病,由ABCC6基因突变所致。弹性纤维性假黄瘤患者有较高的结石发病率且肾内可见斑块形成,ABCC6基因缺陷可能促进Randall斑块的形成。 展开更多
关键词 randall斑块 肾结石 长链非编码RNA ABCC6
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Comparison ofβ-Amyloid Plaque Labeling Methods:Antibody Staining,Gallyas Silver Staining,and Thioflavin-S Staining 被引量:1
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作者 Xinze Shi Xuan Wei +1 位作者 Longze Sha Qi Xu 《Chinese Medical Sciences Journal》 CAS CSCD 2018年第3期167-173,共7页
Objective To evaluate senile plaque formation and compare the sensitivity of three differentβ-amyloid(Aβ)labeling methods(antibody staining,Gallyas silver staining,and thioflavin-S staining)to detect Aβdeposition.M... Objective To evaluate senile plaque formation and compare the sensitivity of three differentβ-amyloid(Aβ)labeling methods(antibody staining,Gallyas silver staining,and thioflavin-S staining)to detect Aβdeposition.Methods APPswe/PSEN1dE9 transgenic mice(APP/PS1)of different ages were used to examine spatiotemporal changes in Aβplaque deposition.Antibody staining,Gallyas silver staining,and thioflavin-S staining were used to detect Aβplaque deposition in the same brain region of adjacent slices from model mice,and the results were compared.Results With aging,Aβplaques first appeared in the cortex and then the deposition increased throughout the whole brain.Significantly greater plaque deposition was detected by 6E10 antibody than that analyzed with Gallyas silver staining or thioflavin-S staining(P<0.05).Plaque deposition did not show significant difference between the APP/PS1 mice brains assayed with Gallyas silver staining and ones with thioflavin-S staining(P=0.0033).Conclusions The APP/PS1 mouse model of Alzheimer’s disease could mimick the progress of Aβplaques occurred in patients with Alzheimer’s disease.Antibody detection of Aβdeposition may be more sensitive than chemical staining methods. 展开更多
关键词 Β-AMYLOID plaques Alzheimer’s disease antibody sTAINING Gallyas silver thioflavin-s
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Randall斑研究进展 被引量:1
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作者 武涛 刘志伟(综述) +1 位作者 唐启胜 马建军(审校) 《医学研究生学报》 CAS 北大核心 2021年第7期770-774,共5页
肾结石的发病率和复发率较高,是泌尿外科的常见病。探索结石形成机制对于预防结石复发至关重要,Randall斑学说是一种被广泛接受的肾结石形成机制,但还没有形成统一的学说。与Randall斑相关的基因研究有助于理解肾结石的形成机制,并为未... 肾结石的发病率和复发率较高,是泌尿外科的常见病。探索结石形成机制对于预防结石复发至关重要,Randall斑学说是一种被广泛接受的肾结石形成机制,但还没有形成统一的学说。与Randall斑相关的基因研究有助于理解肾结石的形成机制,并为未来分子靶向治疗提供方向。Randall斑为临床上肾结石分类提供了一种客观指标,并可能在代谢评估和预测无症状肾结石相关事件发生风险中起到一定作用。文章就Randall斑学说和相关基因研究、在结石分类、代谢评估、无症状肾结石中的应用研究进行综述。 展开更多
关键词 randall 肾结石 基因 分类 无症状肾结石
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肾上皮细胞损伤与肾结石Randall斑块形成 被引量:1
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作者 杨雄 靳潇潇 何文强 《基础医学与临床》 2022年第9期1454-1458,共5页
肾结石具有较高的发病率和复发率,是泌尿外科常见的疾病之一。目前Randall斑块学说是一种被普遍接受的肾结石形成机制,Randall斑块相关研究对肾结石的防治有重要意义,肾上皮细胞损伤促进Randall斑块形成。本文从肾上皮细胞成骨样改变、... 肾结石具有较高的发病率和复发率,是泌尿外科常见的疾病之一。目前Randall斑块学说是一种被普遍接受的肾结石形成机制,Randall斑块相关研究对肾结石的防治有重要意义,肾上皮细胞损伤促进Randall斑块形成。本文从肾上皮细胞成骨样改变、活性氧与氧化应激,巨噬细胞的吞噬,非编码RNA改变的研究进展进行综述。 展开更多
关键词 randall斑块 成骨样改变 巨噬细胞 活性氧 炎性反应
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TMEM16F may be a new therapeutic target for Alzheimer's disease 被引量:4
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作者 Zhi-Qiang Cui Xiao-Ying Hu +6 位作者 Tuo Yang Jing-Wei Guan Ying Gu Hui-Yuan Li Hui-Yu Zhang Qing-Huan Xiao Xiao-Hong Sun 《Neural Regeneration Research》 SCIE CAS CSCD 2023年第3期643-651,共9页
TMEM16F is involved in many physiological processes such as blood coagulation,cell membrane fusion and bone mineralization.Activation of TMEM16F has been studied in various central nervous system diseases.High TMEM16F... TMEM16F is involved in many physiological processes such as blood coagulation,cell membrane fusion and bone mineralization.Activation of TMEM16F has been studied in various central nervous system diseases.High TMEM16F level has been also found to participate in microglial phagocytosis and transformation.Microglia-mediated neuroinflammation is a key factor in promoting the progression of Alzheimer’s disease.However,few studies have examined the effects of TMEM16F on neuroinflammation in Alzheimer’s disease.In this study,we established TMEM16F-knockdown AD model in vitro and in vivo to investigate the underlying regulatory mechanism about TMEM16F-mediated neuroinflammation in AD.We performed a Morris water maze test to evaluate the spatial memory ability of animals and detected markers for the microglia M1/M2 phenotype and NLRP3 inflammasome.Our results showed that TMEM16F was elevated in 9-month-old APP/PS1 mice.After TMEM16F knockdown in mice,spatial memory ability was improved,microglia polarization to the M2 phenotype was promoted,NLRP3 inflammasome activation was inhibited,cell apoptosis and Aβplaque deposition in brain tissue were reduced,and brain injury was alleviated.We used amyloid-beta(Aβ_(25-35))to stimulate human microglia to construct microglia models of Alzheimer’s disease.The levels of TMEM16F,inducible nitric oxide synthase(iNOS),proinflammatory cytokines and NLRP3 inflammasome-associated biomarkers were higher in Aβ_(25-35) treated group compared with that in the control group.TMEM16F knockdown enhanced the expression of the M2 phenotype biomarkers Arg1 and Socs3,reduced the release of proinflammatory factors interleukin-1,interleukin-6 and tumor necrosis factor-α,and inhibited NLRP3 inflammasome activation through reducing downstream proinflammatory factors interleukin-1βand interleukin-18.This inhibitory effect of TMEM16F knockdown on M1 microglia was partially reversed by the NLRP3 agonist Nigericin.Our findings suggest that TMEM16F participates in neuroinflammation in Alzheimer’s disease through participating in polarization of microglia and activation of the NLRP3 inflammasome.These results indicate that TMEM16F inhibition may be a potential therapeutic approach for Alzheimer’s disease treatment. 展开更多
关键词 Alzheimer’s disease plaque inflammatory cytokines M1 phenotype M2 phenotype microglia polarization NEUROINFLAMMATION NLRP3 inflammasome siRNA TMEM16F
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Inhibiting 5-hydroxytryptamine receptor 3 alleviates pathological changes of a mouse model of Alzheimer's disease 被引量:1
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作者 Li-Fen Liu Yu-Tong Liu +5 位作者 Dan-Dan Wu Jie Cheng Na-Na Li Ya-Ni Zheng Liang Huang Qiong-Lan Yuan 《Neural Regeneration Research》 SCIE CAS CSCD 2023年第9期2019-2028,共10页
Extracellular amyloid beta(Aβ) plaques are main pathological feature of Alzheimer’s disease.However,the specific type of neuro ns that produce Aβ peptides in the initial stage of Alzheimer’s disease are unknown.In... Extracellular amyloid beta(Aβ) plaques are main pathological feature of Alzheimer’s disease.However,the specific type of neuro ns that produce Aβ peptides in the initial stage of Alzheimer’s disease are unknown.In this study,we found that 5-hydroxytryptamin receptor 3A subunit(HTR3A) was highly expressed in the brain tissue of transgenic amyloid precursor protein and presenilin-1 mice(an Alzheimer’s disease model) and patients with Alzheimer’s disease.To investigate whether HTR3A-positive interneurons are associated with the production of Aβ plaques,we performed double immunostaining and found that HTR3A-positive interneurons were clustered around Aβ plaques in the mouse model.Some amyloid precursor protein-positive or β-site amyloid precursor protein cleaving enzyme-1-positive neurites near Aβ plaques were co-localized with HTR3A interneurons.These results suggest that HTR3A-positive interneurons may partially contribute to the generation of Aβ peptides.We treated 5.0-5.5-month-old model mice with tro pisetron,a HTR3 antagonist,for 8 consecutive weeks.We found that the cognitive deficit of mice was partially reversed,Aβ plaques and neuroinflammation we re remarkably reduced,the expression of HTR3 was remarkably decreased and the calcineurin/nuclear factor of activated T-cell 4 signaling pathway was inhibited in treated model mice.These findings suggest that HTR3A interneurons partly contribute to generation of Aβ peptide at the initial stage of Alzheimer’s disease and inhibiting HTR3 partly reve rses the pathological changes of Alzheimer’s disease. 展开更多
关键词 5-hydroxytryptamin receptor 3 Alzheimer’s disease amyloid beta plaques CALCINEURIN cognitive deficits HTR3 interneurons iCa2%PLUs% nuclear factor of activated T-cells transgenic amyloid precursor protein and presenilin-1 mice TROPIsETRON
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雷公藤甲素抑制APP/PS1双转基因AD模型小鼠海马内Aβ沉积和老年斑形成 被引量:12
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作者 雷德亮 李明波 +2 位作者 熊鲲 邓小华 罗学港 《解剖学报》 CAS CSCD 北大核心 2009年第3期369-373,共5页
目的研究雷公藤甲素(T10)对APP/PS1双转基因AD模型小鼠(APP/PS1dtg)β-淀粉样蛋白(Aβ)沉积与老年斑(SP)形成的影响。方法取18只4.5月龄健康雄性APP/PS1dtg,随机分为3组,分别以T10灌胃:5μg/(kg·d)(T10 H组)、1μg/(... 目的研究雷公藤甲素(T10)对APP/PS1双转基因AD模型小鼠(APP/PS1dtg)β-淀粉样蛋白(Aβ)沉积与老年斑(SP)形成的影响。方法取18只4.5月龄健康雄性APP/PS1dtg,随机分为3组,分别以T10灌胃:5μg/(kg·d)(T10 H组)、1μg/(kg·d)(T10 L组)和等容量的溶媒灌胃(PLC组),共计45d。45d后取材,左侧半大脑切片,6E10免疫组织化学方法染色和刚果红染色结合无偏性体视学定量分析,研究T10对海马Aβ沉积和SP形成的影响;分离右侧半海马,免疫印迹法分析海马Aβ蛋白水平的变化。结果与PLC组比较,T10 H组海马6E10阳性的Aβ沉积总面积减少了35%(P〈0.001),SP总面积减少了32%(P〈0.001);T10 L组海马Aβ斑总面积减少了18%(P〈0.05),SP总面积减少了21%(P〈0.05);T10呈剂量依赖性抑制Aβ在APP/PS1dtg海马的沉积和SP的形成,减少海马内Aβ蛋白水平;免疫印迹分析也显示,T10 H组海马Aβ蛋白水平最低,PLC组Aβ蛋白水平最高,T10L组Aβ蛋白水平介于两者之间。结论T10抑制Aβ的产生和SP形成,或可用于AD的治疗。 展开更多
关键词 雷公藤甲素 阿尔茨海默病 Β-淀粉样蛋白 老年斑 免疫组织化学 无偏性体视学 转基因小鼠
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Alzheimer′s病研究新进展 被引量:1
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作者 刘欣 郗海涛 刘玉琛 《医学与社会》 2000年第3期19-21,共3页
对近年来国内外有关 AD的病因及发病机理、病理生化及防治等诸方面的研究新进展作一概述。AD的病因及发病机理大致可归纳为环境危险因素学说及遗传危险因素学说 ;病理生化研究方面的进展是 AD患者脑中有神经纤维缠结 (NFT)和老年斑 (SP... 对近年来国内外有关 AD的病因及发病机理、病理生化及防治等诸方面的研究新进展作一概述。AD的病因及发病机理大致可归纳为环境危险因素学说及遗传危险因素学说 ;病理生化研究方面的进展是 AD患者脑中有神经纤维缠结 (NFT)和老年斑 (SP) ,广泛而程度不一的脑萎缩及神经元和突触的缺失以及胆硷能活性降低对其记忆、认知障碍起着重要作用 ;防治研究方面的进展是基因工程研究的迅速进展。考虑到我国国情应当探讨和研究利用中药预防和治疗 AD。 展开更多
关键词 Alzheimer's 神经纤维缠结 老年斑
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人鸟氨酸脱羧酶和S-腺苷甲硫氨酸脱羧酶双反义腺病毒载体的构建
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作者 张冰 刘贤锡 +3 位作者 姜春英 张岩 胡海燕 耿昭 《山东大学学报(医学版)》 CAS 北大核心 2005年第5期387-390,395,共5页
目的:构建人鸟氨酸脱羧酶(ODC)第三外显子和S-腺苷甲硫氨酸脱羧酶(SAMDC)翻译起始位点双反义RNA腺病毒载体。方法:应用RT-PCR方法扩增出SAMDCmRNA翻译起始位点区205bp的基因片断。插入PMD-18T载体中,经XbaI、XhoI酶切回收,反向插入穿梭... 目的:构建人鸟氨酸脱羧酶(ODC)第三外显子和S-腺苷甲硫氨酸脱羧酶(SAMDC)翻译起始位点双反义RNA腺病毒载体。方法:应用RT-PCR方法扩增出SAMDCmRNA翻译起始位点区205bp的基因片断。插入PMD-18T载体中,经XbaI、XhoI酶切回收,反向插入穿梭质粒pAdTrack-CMV-ODCr,形成重组质粒pAdTrack-ODC-SAMDCr。经PmeI酶切线性化后,转入Adeasy-1细菌与pAdeasy-1质粒发生同源重组。挑选阳性重组质粒pAdeasy-ODC-SAMDCr,经PacI酶切后,转染293细胞,包装出腺病毒颗粒,经荧光显微镜和PCR方法对重组腺病毒进行鉴定。结果:利用RT-PCR方法成功地从大肠癌细胞扩增出205bp的cDNA片断,经测序证实为SAMDC翻译起始位点序列。测序证实,重组质粒pAdTrack-ODC-SAMDCr两个基因插入方向和序列均正确,转入Adeasy-1细菌获得多个阳性重组克隆。pAdeasy-ODC-SAMDCr转染293细胞进行包装扩增,可见明显病毒空斑形成,荧光显微镜下可见绿色荧光蛋白在293细胞中表达,PCR法证实含有目的基因。结论:成功构建并扩增出ODC和SAMDC双反义RNA腺病毒载体,为以后肿瘤的基因治疗和预防研究提供了必要工具。 展开更多
关键词 鸟氨酸脱羧酶 s-腺苷甲硫氨酸脱羧酶 腺病毒载体 RNA 反义 病毒空斑
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表面蛋白烯醇化酶Enolase在S.suis 2感染中的角色 被引量:2
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作者 孙雯 潘秀珍 《井冈山大学学报(自然科学版)》 2010年第3期45-48,共4页
目的通过克隆表达,在获得具有酶活性的猪链球菌2型(S.suis2)05ZYH33重组烯醇化酶Enolase蛋白的基础上,旨在继续探索其在细菌粘附和引发免疫下调作用中的角色。方法流式细胞术(FCM)、Hep2细胞粘附竞争试验、免疫空斑试验。结果流式细胞术... 目的通过克隆表达,在获得具有酶活性的猪链球菌2型(S.suis2)05ZYH33重组烯醇化酶Enolase蛋白的基础上,旨在继续探索其在细菌粘附和引发免疫下调作用中的角色。方法流式细胞术(FCM)、Hep2细胞粘附竞争试验、免疫空斑试验。结果流式细胞术FCM的细胞定位显示Enolase可以部分存在S.suis205ZYH33细菌的表面;Hep2细胞粘附竞争试验表明猪链球菌表面Enolase参与细菌对宿主细胞的黏附作用;免疫空斑实验的结果揭示Enolase在抑制宿主特异性免疫应答中发挥作用。结论 Enolase作为一个表面蛋白,确实在S.suis2感染中扮演一定的角色。 展开更多
关键词 猪链球菌2型 烯醇化酶Enolase 流式细胞术 黏附 溶血空斑形成实验
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脑卒中高危人群颈动脉斑块与血清sCD40L之间的关系 被引量:5
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作者 秋大成 侯丽淳 +3 位作者 王辰 毕胜 杨慧 王复新 《中风与神经疾病杂志》 CAS 北大核心 2015年第10期922-925,共4页
目的观察血清可溶性CD40配体(s CD40L)与高危人群颈动脉斑块性质之间的关系,为高危人群早期服用阿司匹林预防缺血性脑血管病提供依据。方法根据入选条件选取145例作为高危组,选取健康体检者40例作为对照组,据颈动脉超声检查结果将高危... 目的观察血清可溶性CD40配体(s CD40L)与高危人群颈动脉斑块性质之间的关系,为高危人群早期服用阿司匹林预防缺血性脑血管病提供依据。方法根据入选条件选取145例作为高危组,选取健康体检者40例作为对照组,据颈动脉超声检查结果将高危组分为无斑块组(Ⅰ)67例、稳定斑块组(Ⅱ)21例和不稳定斑块组(Ⅲ)57例3个亚组,采用酶联免疫吸附法(ELISA)测定所有研究对象血清s CD40L水平。结果高危组血清s CD40L水平明显高于对照组,差异有统计学意义P<0.05;高危组各亚组间血清s CD40L水平比较:Ⅰ组<Ⅱ组<Ⅲ组[(3.489±0.820)mg/ml<(3.600±0.697)mg/ml<(4.011±1.074)mg/ml],差异有统计学意义P<0.05;经Logistic回归分析结果显示:血清s CD40L(OR=1.788,P=0.037)是影响颈动脉斑块性质的危险因素。结论血清s CD40L是颈动脉粥样硬化斑块形成的危险因素;血清s CD40L可能影响颈动脉粥样硬化斑块的性质,促使其由稳定斑块向不稳定斑块的方向发展;对于脑卒中高危人群血清s CD40L水平较高者,应早期给予阿司匹林药物治疗,以此来预防缺血性脑血管病的发生、发展。 展开更多
关键词 sCD40L 动脉粥样硬化 斑块不稳定性 脑卒中
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APP/PS1/LC3三转基因小鼠的构建及其自噬流水平研究 被引量:2
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作者 陈婧菲 贺桂琼 龙志敏 《重庆医科大学学报》 CAS CSCD 北大核心 2018年第9期1140-1146,共7页
目的:研究APP/PS1/LC3三转基因小鼠的构建及自噬流情况。方法:将CAG-m RFP-e GFP-LC3转基因自噬流模型小鼠与APP/PS1双转基因阿尔茨海默病(Alzheimer’s Disease,AD)模型小鼠交配繁殖,对产下的子代进行基因分型鉴定,选出同时含有CAG-m R... 目的:研究APP/PS1/LC3三转基因小鼠的构建及自噬流情况。方法:将CAG-m RFP-e GFP-LC3转基因自噬流模型小鼠与APP/PS1双转基因阿尔茨海默病(Alzheimer’s Disease,AD)模型小鼠交配繁殖,对产下的子代进行基因分型鉴定,选出同时含有CAG-m RFP-e GFP-LC3基因和APP/PS1的小鼠为APP/PS1/LC3三转基因小鼠,Morris水迷宫实验检测小鼠行为学变化。取6月龄APP/PS1/LC3三转基因小鼠和同窝CAG-m RFP-e GFP-LC3单转基因小鼠各6只,断头取脑后在透射电镜及荧光显微镜下观察自噬流的变化,采用免疫组织化学法检测老年斑的形成,用Western blot检测自噬标志物。结果:基因分型证实APP/PS1/LC3三转基因小鼠构建成功。Morris水迷宫显示,6月龄APP/PS1/LC3三转基因小鼠找到平台的平均潜伏期和路程均明显增加(F=87.096,P=0.000;F=41.583,P=0.000),穿越平台的次数明显减少(2.000±0.707 vs.4.800±0.800,t=2.622,P=0.031)。透射电镜下观察,6月龄APP/PS1/LC3三转基因小鼠脑神经元内出现更多的自噬小泡。荧光显微镜下观察,与同窝CAG-m RFPe GFP-LC3单转基因小鼠比较,6月龄APP/PS1/LC3三转基因小鼠海马区自噬体及自噬溶酶体数量均明显增高(5.894±0.742 vs.14.820±3.350,t=0.017,P=0.000;1.204±0.420 vs.1.840±0.559,t=3.156,P=0.005),大脑皮质内自噬体数量亦明显升高(1.943±0.415 vs.10.030±4.382,t=6.364,P=0.000),但自噬溶酶体数量比较,差异未见统计学意义(0.562±0.207 vs.0.686±0.195,t=0.156,P=0.878)。免疫组化染色结果显6月龄APP/PS1/LC3三转基因大脑皮质及海马区域出现明显老年斑,而同窝CAG-m RFP-e GFP-LC3单转基因小鼠脑内未见老年斑。Western blot结果显示:与同窝CAG-m RFP-e GFP-LC3单转基因小鼠比较,APP/PS1/LC3三转基因小鼠脑内APP蛋白明显升高(0.294±0.070 vs.0.690±0.275,t=3.423,P=0.007),自噬相关蛋白LC3、Beclin1、P62水平均增高(0.241±0.004 vs.0.534±0.019,t=37.170,P=0.000;0.479±0.020 vs.1.180±0.255,t=6.820,P=0.000;0.188±0.007 vs.0.356±0.021,t=18.850,P=0.000),但溶酶体膜蛋白LAMP1表达水平降低(1.450±0.065 vs.0.773±0.043,t=8.705,P=0.000)。结论:APP/PS1/LC3三转基因小鼠自噬被激活,但自噬溶酶体降解受阻,是研究AD自噬流水平的理想动物模型。 展开更多
关键词 阿尔茨海默病 自噬 老年斑 转基因小鼠
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急性脑梗死患者Cathepsin S/Cystatin C水平与动脉斑块性质及狭窄程度的相关性分析 被引量:13
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作者 韩新生 徐建可 +4 位作者 李相磊 张洪阳 韩苗 张蕴 许予明 《中风与神经疾病杂志》 CAS 2018年第5期434-438,共5页
目的探讨急性脑梗死患者血清组织蛋白酶S(Cat S)与胱抑素C(Cys C)表达水平与动脉斑块性质及狭窄程度的关系。方法 336例急性脑梗死患者根据斑块性质及血管狭窄程度分为:稳定斑块组与不稳定斑块组及轻度、中度和重度狭窄3个亚组;114例同... 目的探讨急性脑梗死患者血清组织蛋白酶S(Cat S)与胱抑素C(Cys C)表达水平与动脉斑块性质及狭窄程度的关系。方法 336例急性脑梗死患者根据斑块性质及血管狭窄程度分为:稳定斑块组与不稳定斑块组及轻度、中度和重度狭窄3个亚组;114例同期健康体检者为正常对照组。所有入组患者检测血清Cat S和Cys C,脑梗死患者行神经功能缺损评价。结果脑梗死组患者血清Cat S及Cat S/Cys C均高于对照组,而血清Cys C水平降低,Cat S分别为(63.07±20.81)pg/ml、(50.49±10.54)pg/ml,Cat S/Cys C分别为(78.83±6.88)、(47.18±3.32),Cys C分别为(0.80±0.11)mg/L、(1.07±0.83)mg/L,两组比较,差异均有统计学意义(P<0.05);不稳定斑块组Cat S及Cat S/Cys C较稳定斑块组增高,而血清Cys C水平较稳定斑块组降低,Cat S分别为(75.34±15.45)pg/ml、(60.12±18.53)pg/ml,Cat S/Cys C分别为(103.68±2.52)、(64.64±9.24),Cys C分别为(0.73±0.62)mg/L、(0.93±0.53)mg/L,两组比较,差异均有统计学意义(P<0.05);两组亚组间(轻度狭窄组、中度狭窄组及重度狭窄组)比较,Cat S、Cys C水平及Cat S/Cys C均有显著差异(P<0.05),而血管狭窄程度越重,Cat S及Cat S/Cys C增高越明显、Cys C降低越明显,差异均有统计学意义(P<0.01)。结论脑梗死患者颈动脉斑块稳定性越差、血管狭窄程度越重,血清Cat S水平就越高,呈正相关;血清Cat S及Cat S/Cys C增高和血清Cys C水平降低,与脑梗死的发生密切相关。 展开更多
关键词 组织蛋白酶/ 胱抑素C 斑块 血管狭窄 缺血性脑卒中
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Surgery for Peyronie's disease 被引量:11
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作者 Laurence A Levine Stephen M Larsen 《Asian Journal of Andrology》 SCIE CAS CSCD 2013年第1期27-34,共8页
Peymnie's disease (PD) is most simply referred to as a fibrotic wound-healing disorder of the tunica albuginea. It is both a physically and psychologically devastating disorder that causes penile deformity, curvatu... Peymnie's disease (PD) is most simply referred to as a fibrotic wound-healing disorder of the tunica albuginea. It is both a physically and psychologically devastating disorder that causes penile deformity, curvature, hinging, narrowing and shortening, which may compromise sexual function. Although a variety of non-surgical treatments have been suggested, none to date offer a reliable and effective correction of the penile deformity. As a result, surgery remains the gold standard treatment option, offering the most rapid and reliable treatment which will be the focus of this article. We review the preoperative evaluation, surgical algorithm, graft materials and postoperative management of PD. Outcomes for tunical shortening, tunical lengthening and penile prosthesis placement for penile straightening are reviewed. Tunica albuginea plication is the preferred method of straightening for men with adequate rigidity and less severe disease defined as curvature less than 70° without narrowing/hinging. Men who have more severe, complex deformity, but maintain strong preoperative erectile function should be considered candidates for straightening with plaque incision or partial excision and grafting. Finally, for those men who have inadequate rigidity and PD, penile prosthesis placement with straightening is the best approach to address both problems. 展开更多
关键词 penile prosthesis Peyronie's disease penile reconstruction plaque excision and grafting plaque incision tunica albuginea plication
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Alzheimer's disease: the silver tsunami of the 21^(st) century 被引量:5
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作者 Ankita Sarkar Madison Irwin +2 位作者 Aditi Singh Matthew Riccetti Amit Singh 《Neural Regeneration Research》 SCIE CAS CSCD 2016年第5期693-697,共5页
Alzheimer's disease(AD), a fatal progressive neurodegenerative disorder, has no cure to date. One of the causes of AD is the accumulation of amyloid-beta 42(Aβ42) plaques, which result in the onset of neurodegen... Alzheimer's disease(AD), a fatal progressive neurodegenerative disorder, has no cure to date. One of the causes of AD is the accumulation of amyloid-beta 42(Aβ42) plaques, which result in the onset of neurodegeneration. It is not known how these plaques trigger the onset of neurodegeneration. There are several animal models developed to(i) study etiology of disease,(ii) look for genetic modifiers, and(iii) identify chemical inhibitors that can block neurodegeneration and help to find cure for this disease. An insect model of Drosophila melanogaster has also provided new insights into the disease. Here we will discuss the utility of the Drosophila eye model to study Alzheimer's disease. 展开更多
关键词 Alzheimer's disease NEURODEGENERATION amyloid plaques amyloid hypothesis animal models drosophila melanogaster drosophila eye
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Natural polyphenols effects on protein aggregates in Alzheimer's and Parkinson's prion-like diseases 被引量:3
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作者 aline freyssin guylène page +1 位作者 bernard fauconneau agnès rioux bilan 《Neural Regeneration Research》 SCIE CAS CSCD 2018年第6期955-961,共7页
Alzheimer's and Parkinson's diseases are the most common neurodegenerative diseases. They are characterized by protein aggregates and so can be considered as prion-like disease. The major components of these deposit... Alzheimer's and Parkinson's diseases are the most common neurodegenerative diseases. They are characterized by protein aggregates and so can be considered as prion-like disease. The major components of these deposits are amyloid peptide and tau for Alzheimer's disease, α-synuclein and synphilin-1 for Parkinson's disease. Drugs currently proposed to treat these pathologies do not prevent neurodegenerative processes and are mainly symptomatic therapies. Molecules inducing inhibition of aggregation or disaggregation of these proteins could have beneficial effects, especially if they have other beneficial effects for these diseases. Thus, several natural polyphenols, which have antioxidative, anti-inflammatory and neuroprotective properties, have been largely studied, for their effects on protein aggregates found in these diseases, notably in vitro. In this article, we propose to review the significant papers concerning the role of polyphenols on aggregation and disaggregation of amyloid peptide, tau, α-synuclein, synphilin-1, suggesting that these compounds could be useful in the treatments in Alzheimer's and Parkinson's diseases. 展开更多
关键词 natural polyphenols protein aggregates Alzheimer's disease amyloid peptide amyloid plaques hyperphosphorylated tau Parkinson's disease a-synuclein synphilin-I
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