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CRISPR-Cas9基因编辑技术对细胞内源蛋白进行荧光标记的实验操作 被引量:2
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作者 吴仲胜 高誉 +2 位作者 杜勇涛 党颂 何康敏 《遗传》 CAS CSCD 北大核心 2023年第2期165-175,共11页
CRISPR-Cas9是目前广泛应用的基因编辑技术,可对目的基因进行高效精准编辑,快速实现目的基因的敲除或敲入。Cas9蛋白在sgRNA引导下对靶序列进行剪切并造成DNA双链断裂,在与剪切位点两端同源的DNA模板序列存在时,可通过同源重组修复方式... CRISPR-Cas9是目前广泛应用的基因编辑技术,可对目的基因进行高效精准编辑,快速实现目的基因的敲除或敲入。Cas9蛋白在sgRNA引导下对靶序列进行剪切并造成DNA双链断裂,在与剪切位点两端同源的DNA模板序列存在时,可通过同源重组修复方式引入外源序列,实现荧光蛋白或其他标签在基因组上的精准敲入,进而实现对内源蛋白进行荧光标签的融合标记。通过基因编辑技术对内源目的蛋白进行标记,可避免由于过表达造成蛋白质定位、动力学或功能等的潜在影响,可显著提升细胞成像实验的稳定性和可重复性。本文重点介绍了利用CRISPR-Cas9基因编辑系统对目的蛋白进行荧光蛋白或自标记蛋白标签标记的方法与操作流程,为构建内源蛋白荧光标记的哺乳动物细胞系提供参考。 展开更多
关键词 CRISPR-Cas9基因编辑 活细胞 内源蛋白 荧光蛋白 自标记蛋白标签
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Effects of neuregulin-1 on autonomic nervous system remodeling post-myocardial infarction in a rat model 被引量:7
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作者 Xin Lai Liang Zhong +7 位作者 Hai-xia Fu song dang Xin Wang Ning Zhang Gao-ke Feng Zi-qiang Liu Xi Wang Long Wang 《Neural Regeneration Research》 SCIE CAS CSCD 2017年第11期1905-1910,共6页
Sympathetic nerve and vagus nerve remodeling play an important part in cardiac function post-myocardial infarction (MI). Increasing evidence indicates that neuregulin-1 (NRG-1) improves cardiac function following ... Sympathetic nerve and vagus nerve remodeling play an important part in cardiac function post-myocardial infarction (MI). Increasing evidence indicates that neuregulin-1 (NRG-1) improves cardiac function following heart failure. Since its impact on cardiac function and neural remodeling post-MI is poorly understood, we aimed to investigate the role of NRG-1 in autonomic nervous system remodeling post-MI. Forty-five Sprague-Dawley rats were equally randomized into three groups: sham (with the left anterior descending coronary artery exposed but without ligation), MI (left anterior descending coronary artery ligation), and MI plus NRG-1 (left anterior descending coronary artery ligation followed by intraperitoneal injection of NRG-1 (10 lag/kg, once daily for 7 days)). At 4 weeks after MI, echocardi- ography was used to detect the rat cardiac function by measuring the left ventricular end-systolic inner diameter, left ventricular diastolic diameter, left ventricular end-systolic volume, left ventricular end-diastolic volume, left ventricular ejection fraction, and left ventricular fractional shortening, mRNA and protein expression levels of tyrosine hydroxylase, growth associated protein-43 (neuronal specific pro- tein), nerve growth factor, choline acetyltransferase (vagus nerve marker), and vesicular acetylcholine transporter (cardiac vagal nerve fiber marker) in ischemic myocardia were detected by real-time PCR and western blot assay to assess autonomous nervous remodeling. After MI, the rat cardiac function deteriorated significantly, and it was significantly improved after NRG-1 injection. Compared with the MI group, mRNA and protein levels of tyrosine hydroxylase and growth associated protein-43, as well as choline acetyltransferase mRNA level significantly decreased in the MI plus NRG-1 group, while mRNA and protein levels of nerve growth factor and vesicular acetylcholine transporters, as well as choline acetyltransferase protein level slightly decreased. Our results indicate that NRG- 1 can improve cardiac function and regulate sympathetic and vagus nerve remodeling post-MI, thus reaching a new balance of the autonomic nervous system to protect the heart from injury. 展开更多
关键词 nerve remodeling myocardial infarction NEUREGULIN-1 sympathetic nerve vagus nerve animal model real-time PCR westernblot assay cardiac function ECHOCARDIOGRAPHY
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Effects of Traditional Chinese Medicine Shensong Yangxin Capsules on Heart Rhythm and Function in Congestive Heart Failure Patients with Frequent Ventricular Premature Complexes:A Randomized, Double-blind, Multicenter Clinical Trial 被引量:36
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作者 Xi Wang Dan HU +9 位作者 song dang He Huang Cong-Xin Huang Ming-Jie Yuan Yan-Hong Tang Qing-Shan Zheng Fang Yin Shu Zhang Bo-Li Zhang Run-Lin Gao 《Chinese Medical Journal》 SCIE CAS CSCD 2017年第14期1639-1647,共9页
Background:Pharmacological therapy for congestive heart failure (CHF) with ventricular arrhythmia is limited.In the study,our aim was to evaluate the effects of Chinese traditional medicine Shensong Yangxin capsul... Background:Pharmacological therapy for congestive heart failure (CHF) with ventricular arrhythmia is limited.In the study,our aim was to evaluate the effects of Chinese traditional medicine Shensong Yangxin capsules (SSYX) on heart rhythm and function in CHF patients with frequent ventricular premature complexes (VPCs).Methods:This double-blind,placebo-controlled,multicenter study randomized 465 CHF patients with frequent VPCs to the SSYX (n =232) and placebo groups (n =233) for 12 weeks of treatment.The primary endpoint was the VPCs monitored by a 24-h ambulatory electrocardiogram.The secondary endpoints included the left ventricular ejection fraction (LVEF),left ventricular end-diastolic diameter,N-terminal pro-brain natriuretic peptide (NT-proBNP),New York Heart Association (NYHA) classification,6-min walking distance (6MWD),Minnesota Living with Heart Failure Questionnaire (MLHFQ) scores,and composite cardiac events (CCEs).Results:The clinical characteristics were similar at baseline.SSYX caused a significantly greater decline in the total number of VPCs than the placebo did (-2145 ± 2848 vs.-841 ± 3411,P 〈 0.05).The secondary endpoints of the LVEE NYHA classification,NT-proBNP,6MWD,and MLHFQ scores showed a greater improvements in the SSYX group than in the placebo group (ALVEF at 12th week:4.75 ± 7.13 vs.3.30 ± 6.53;NYHA improvement rate at the 8th and 12th week:32.6% vs.21.8%,40.5% vs.25.7%;mean level of NT-proBNP in patients with NT-proBNP 〉125 pg/ml at 12th week:-122 [Q1,Q3:-524,0] vs.-75 [Q1,Q3:-245,0];A6MWD at 12th week:35.1 ± 38.6 vs.17.2 ± 45.6;AMLHFQ at the 4th,8th,and 12th week:-4.24 ± 6.15 vs.-2.31 ± 6.96,-8.l 9 ± 8.41 vs.-3.25 ± 9.40,10.60 ± 9.41 vs.-4.83 ± 11.23,all P 〈 0.05).CCEs were not different between the groups during the study period.Conclusions:In this 12-week pilot study,SSYX was demonstrated to have the benefits of VPCs suppression and cardiac function improvement with good compliance on a background of standard treatment for CHF. 展开更多
关键词 Congestive Heart Failure Randomized ControlledTrial Shensong Yangxin Capsules Ventricular PrematureComplexes
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A multi-responsive luminescent sensor towards Fe_(3+) and acetone based on a Cd-containing metal–organic framework 被引量:5
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作者 Ting Wang Qing-Hui Liu +4 位作者 Ying Gao Xiu-Yun Yang Weiting Yang song dang Zhong-Ming Sun 《Chinese Chemical Letters》 SCIE CAS CSCD 2016年第4期497-501,共5页
A Cd-containing metal–organic framework(Cd L), formula as {[Cd_3(L)_2(H_2O)_6] 1.5DMF}, has been synthesized under solvothermal condition by the reaction of 4,40,400-(methylsilanetriyl)tribenzoic acid(H_3L)... A Cd-containing metal–organic framework(Cd L), formula as {[Cd_3(L)_2(H_2O)_6] 1.5DMF}, has been synthesized under solvothermal condition by the reaction of 4,40,400-(methylsilanetriyl)tribenzoic acid(H_3L) and Cd^(2+)ion. Single-crystal X-ray diffraction reveals that Cd L displays a three-dimensional framework with 2-fold interpenetration and DMF molecules locate in the void space of the channels. A topological analysis of the framework indicates Cd Lisa 3,4-connected pto net. The photoluminescence properties of Cd L are systematically studied in detail. Impressively, Cd L shows excellent detection performance towards Fe^(3+)ion and acetone in the sensing experiments, which undoubtedly demonstrates the great potential of Cd L as a highly selective multi-responsive luminescent sensor for the detection of organic solvents and metal ions. 展开更多
关键词 Metal–organic framework Fluorescent probe Multi-responsive sensor Luminescent quenching
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Microwave-assisted synthesis of nanoscale Eu(BTC)(H_2O)·DMF with tunable luminescence 被引量:3
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作者 song dang Shuyan song +1 位作者 Jing Feng Hongjie Zhang 《Science China Chemistry》 SCIE EI CAS CSCD 2015年第6期973-978,共6页
Nanoscale europium(III) metal-organic frameworks, Eu(BTC)(H2O)?DMF, were synthesized by rapid microwave-assisted method. The components of the as-prepared products were confirmed by the elemental analysis, X-ray powde... Nanoscale europium(III) metal-organic frameworks, Eu(BTC)(H2O)?DMF, were synthesized by rapid microwave-assisted method. The components of the as-prepared products were confirmed by the elemental analysis, X-ray powder diffraction(XRD), thermal gravimetric analysis(TGA) and Fourier-transform infrared spectra(FTIR) analyses. Eu(BTC)(H2O)?DMF with various morphologies, including particle-like, rod-like, straw-sheaf-like nanostructures, could be simply prepared by controlling the concentrations of the starting reactants. The optical measurements on the obtained Eu(BTC)(H2O)?DMF indicated that all the nanomaterials show the characteristic emissions of the Eu3+ ions at 578, 590, 612, 650, and 699 nm, which were attributed to 5D0?7FJ(J=0–4) transitions of the Eu3+ ion, respectively. It was also noticed that the luminescent properties of the as-prepared products were heavily dependent on the morphologies and sizes of the nanomaterials. The assembled straw-sheaf-like architectures displayed the strongest emissions and the longest luminescence lifetime, which was mainly due to the fewest surface defects. 展开更多
关键词 微波辅助合成 纳米结构 发光性能 铕离子 BTC DMF 傅里叶变换红外光谱 粉末X射线衍射
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Lanthanide complexes-functionalized ordered mesoporous TiO_2:Multicolor emission(visible and near-infrared luminescence) based on visible-light sensitization 被引量:1
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作者 Zhijuan Wang song dang +1 位作者 Shu'na Zhao Lining Sun 《Journal of Rare Earths》 SCIE EI CAS CSCD 2018年第9期939-945,共7页
Recently,much attention has been paid to the lanthanide luminescent materials based on the visiblelight sensitization for their potential applications in the fields of bio-imaging and optical devices.In this work,the ... Recently,much attention has been paid to the lanthanide luminescent materials based on the visiblelight sensitization for their potential applications in the fields of bio-imaging and optical devices.In this work,the lanthanide complexes have been covalently bonded to the ordered mesoporous titania(OMT) matrix,and the resulting titania-based hybrid ordered mesoporous materials(named as LnDBOMT,Ln = Eu,Sm,Yb,Nd) were characterized by using Fourier-transform infrared(FT-IR) spectroscopy,small-angle X-ray powder diffraction(SAXD),N2 adsorption-desorption isotherms,transmission electron microscopy(TEM),fluorescence spectroscopy,and thermogravimetric analysis.Generally,exciting with visible light is advantageous over UV excitation.Of importance here is that,under excitation with visible light,the LnDB-OMT all show characteristic visible(Eu3+,Sm3+) as well as nearinfrared(Sm3+,Yb3+,Nd3+) luminescence of the corresponding Ln3+ ions(multicolor emission covered from 500 to 1400 nm spectral region),which is attributed to the energy transfer from the ligands to the Ln3+ ions via an antenna effect. 展开更多
关键词 Visible-light excitation Visible luminescence Near-infrared luminescence Lanthanide complexes-functionalized Ordered mesoporous titania Rare earths
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