目的用Meta分析的方法分析TGF-β3基因多态性与亚洲人群非综合征型唇腭裂(NSCL/P)易感性的相关性。方法计算机检索PubMed、Cochrane Library、Embase、中文科技期刊全文数据库、中国期刊全文数据库、中国生物医学文献数据库和万方数据库...目的用Meta分析的方法分析TGF-β3基因多态性与亚洲人群非综合征型唇腭裂(NSCL/P)易感性的相关性。方法计算机检索PubMed、Cochrane Library、Embase、中文科技期刊全文数据库、中国期刊全文数据库、中国生物医学文献数据库和万方数据库,检索日期自建库至2019年1月31日公开发表的文献。2位研究者独立按照本文纳入和排除标准筛选文献、提取资料。采用Stata12.0软件进行Meta分析,漏斗图观察发表偏倚。结果9篇病例对照研究进入Meta分析。TGF-β3基因在等位基因模型(G vs A:OR=1.23,95%CI:1.04~1.47)、相加模型(GG vs AA:OR=1.72,95%CI:1.17~2.55)、共显性模型(GG vs GA:OR=1.50,95%CI:1.15~1.98)和显性模型(GG vs GA+AA:OR=1.50,95%CI:1.16~1.94)下与NSCL/P易感性有关;在隐性模型(AA vs GA+GG:OR=0.97,95%CI:0.72~1.30)二者无相关性。不同对照组人群来源亚组分析结果显示,对照组来源医院的人群,各种基因模型下差异均无统计学意义。对照来源社区人群,TGF-β3基因位点多态性在A vs G、GA vs GG和GA+AA vs GG下与NSCL/P易感性有关,AA vs GG和AA vs GG+GA无相关性。结论TGF-β3基因多态性增加了亚洲人群NSCL/P的发病风险。展开更多
目的探讨TGF-β3、透明质酸(hyaluronic acid,HA)、甲状旁腺激素相关蛋白(parathyroid hormonerelated protein,PTHrP)对人脐带间充质干细胞(human umbilical cord mesenchymal stem cells,HUC-MSCs)成软骨分化的影响,优化培养条件的组...目的探讨TGF-β3、透明质酸(hyaluronic acid,HA)、甲状旁腺激素相关蛋白(parathyroid hormonerelated protein,PTHrP)对人脐带间充质干细胞(human umbilical cord mesenchymal stem cells,HUC-MSCs)成软骨分化的影响,优化培养条件的组合方式。方法体外分离培养人脐带间充质干细胞,形态学观察,流式检测表面抗体及成脂、成骨分化鉴定。取P3代细胞,加入不同组合的细胞因子进行成软骨诱导培养,分为1~21 d TGF-β3(G1)、1~28 d TGF-β3(G2)、1~21 d TGF-β3/1~21 d HA(G3)、1~28 d TGF-β3/1~28 d HA(G4)、1~21 d TGF-β3/1~21 d HA、14~21 d PTHrP(G5)、1~28 d TGF-β3、1~28 d HA、14~28 d PTHrP(G6)、1~28 d TGF-β3、1~28 d HA、21~28 d PTHrP(G7) 7组。结果诱导培养28 d和21 d相比较,TGF-β3组和TGFβ3/HA组COL2α1、COL10α1、SOX9、ACAN的表达量上升;TGFβ3/HA/PTHrP组COL2α1、SOX9、ACAN的表达量上升,COL10α1下降。培养相同天数时,TGFβ3/HA组较TGF-β3组COL2α1、SOX9、ACAN的表达量上升;TGFβ3/HA/PTHrP组同TGF-β3/HA组比较,COL2α1、ACAN、SOX9的表达量增加,COL10α1表达量下降,且PTHrP加入2周较1周效果更明显。差异均有统计学意义。HE和阿利新蓝染色结果与实时荧光定量PCR的结果一致。结论 TGF-β3、HA、PTHrP可以不同程度的促软骨分化,其中联合应用TGF-β3,HA 28 d,在第14天时加入PTHrP一起培养,这种组合方式促进HUC-MSCs成软骨分化效果最好,且能抑制细胞肥大。展开更多
Summary： A new type of TGF-β3 fusion protein with targeted therapy function was constructed, and its feasibility and target specificity of inducing chondrogenesis were investigated by transfecting LAP-MMP-mTGF-β3 g...Summary： A new type of TGF-β3 fusion protein with targeted therapy function was constructed, and its feasibility and target specificity of inducing chondrogenesis were investigated by transfecting LAP-MMP-mTGF-β3 gene into adipose-derived stem cells （ADSCs）. The recombinant pIRES- EGFP-MMP was constructed by inserting the sense and antisense DNA of encoding the amino acid of the synthetic MMP enzyme cutting site into the eukaryotic expression vector pIRES-EGFE LAP and mTGF-β3 fragments were obtained by using RT-PCR and inserted into the upstream and downstream of MMP from pIRES-EGFP-MMP respectively, and the recombinant plasmid of pIRES-EGFP- LAP-MMP-mTGF-β3 was constructed, which was transferred to ADSCs. The ADSCs were cultured and divided in three groups： experimental group （MMP group）, negative control group （no MMP） and non-transfection group. The morphological changes were observed microscopically, and the expression of proteoglycan and type II collagen （Col II） was detected by using Alcian blue staining and immuno- histochemistry staining at 7th, 14th and 21st day after culture. The recombinant plasmid of pIRES-EGFP-LAP-MMP-mTGF-β3 was correctly constructed by methods of enzyme cutting and se- quencing analysis. The mTGF-β3 fusion protein was successfully expressed after transfection, and in the presence of the MMP, active protein mTGF-β3 was generated, which significantly promoted differ- entiation of ADSCs into chondrocytes and the expression of cartilage matrix. The novel fusion protein LAP-MMP-mTGF-β3 can targetedly induce differentiation of ADSCs into chondrocytes, which would open up prospects for target therapy of cartilage damage repair in future.展开更多
文摘为了研究转化生长因子β3(Transforming growth factor beta3,TGF-β3)对体外培养的羊驼皮肤黑色素细胞表型的影响。本研究在体外培养的羊驼皮肤黑色素细胞中添加不同浓度TGF-β3(6.25、12.5、25、50ng·mL-1),通过实时监测和检测细胞增殖、毛色相关基因小眼畸形相关转录因子(Microphthalmia—associtated transcription factor,MITF)、酪氨酸酶(Tyrosinase,TYR)和酪氨酸酶相关蛋白2(Tyrosinase related protein 2,TYRP2)表达以及黑色素产量的变化。结果表明:(1)在羊驼皮肤黑色素细胞中添加50ng·mL-1浓度的TGF-β3后,在前30h内对细胞增殖有抑制效果,30h后对细胞增殖有明显的长时程维持细胞数量作用,但对TGF-β3添加的剂量没有依赖性;(2)添加TGF-β3后,黑色素细胞内MITF、TYR和TYRP2的表达量均被下调,而且黑色素细胞产生黑色素的量也被下调,主要以添加50ng·mL-1时下调最为显著。结果揭示,TGF-β3通过对羊驼黑色素细胞内MITF、TYR和TYRP2的表达的影响,并调控黑色素的产生,对黑色素细胞的生物学功能具有重要的影响。
文摘目的用Meta分析的方法分析TGF-β3基因多态性与亚洲人群非综合征型唇腭裂(NSCL/P)易感性的相关性。方法计算机检索PubMed、Cochrane Library、Embase、中文科技期刊全文数据库、中国期刊全文数据库、中国生物医学文献数据库和万方数据库,检索日期自建库至2019年1月31日公开发表的文献。2位研究者独立按照本文纳入和排除标准筛选文献、提取资料。采用Stata12.0软件进行Meta分析,漏斗图观察发表偏倚。结果9篇病例对照研究进入Meta分析。TGF-β3基因在等位基因模型(G vs A:OR=1.23,95%CI:1.04~1.47)、相加模型(GG vs AA:OR=1.72,95%CI:1.17~2.55)、共显性模型(GG vs GA:OR=1.50,95%CI:1.15~1.98)和显性模型(GG vs GA+AA:OR=1.50,95%CI:1.16~1.94)下与NSCL/P易感性有关;在隐性模型(AA vs GA+GG:OR=0.97,95%CI:0.72~1.30)二者无相关性。不同对照组人群来源亚组分析结果显示,对照组来源医院的人群,各种基因模型下差异均无统计学意义。对照来源社区人群,TGF-β3基因位点多态性在A vs G、GA vs GG和GA+AA vs GG下与NSCL/P易感性有关,AA vs GG和AA vs GG+GA无相关性。结论TGF-β3基因多态性增加了亚洲人群NSCL/P的发病风险。
文摘目的探讨TGF-β3、透明质酸(hyaluronic acid,HA)、甲状旁腺激素相关蛋白(parathyroid hormonerelated protein,PTHrP)对人脐带间充质干细胞(human umbilical cord mesenchymal stem cells,HUC-MSCs)成软骨分化的影响,优化培养条件的组合方式。方法体外分离培养人脐带间充质干细胞,形态学观察,流式检测表面抗体及成脂、成骨分化鉴定。取P3代细胞,加入不同组合的细胞因子进行成软骨诱导培养,分为1~21 d TGF-β3(G1)、1~28 d TGF-β3(G2)、1~21 d TGF-β3/1~21 d HA(G3)、1~28 d TGF-β3/1~28 d HA(G4)、1~21 d TGF-β3/1~21 d HA、14~21 d PTHrP(G5)、1~28 d TGF-β3、1~28 d HA、14~28 d PTHrP(G6)、1~28 d TGF-β3、1~28 d HA、21~28 d PTHrP(G7) 7组。结果诱导培养28 d和21 d相比较,TGF-β3组和TGFβ3/HA组COL2α1、COL10α1、SOX9、ACAN的表达量上升;TGFβ3/HA/PTHrP组COL2α1、SOX9、ACAN的表达量上升,COL10α1下降。培养相同天数时,TGFβ3/HA组较TGF-β3组COL2α1、SOX9、ACAN的表达量上升;TGFβ3/HA/PTHrP组同TGF-β3/HA组比较,COL2α1、ACAN、SOX9的表达量增加,COL10α1表达量下降,且PTHrP加入2周较1周效果更明显。差异均有统计学意义。HE和阿利新蓝染色结果与实时荧光定量PCR的结果一致。结论 TGF-β3、HA、PTHrP可以不同程度的促软骨分化,其中联合应用TGF-β3,HA 28 d,在第14天时加入PTHrP一起培养,这种组合方式促进HUC-MSCs成软骨分化效果最好,且能抑制细胞肥大。
基金supported by the National Natural Science Foundation of China(No.81101376)
文摘Summary： A new type of TGF-β3 fusion protein with targeted therapy function was constructed, and its feasibility and target specificity of inducing chondrogenesis were investigated by transfecting LAP-MMP-mTGF-β3 gene into adipose-derived stem cells （ADSCs）. The recombinant pIRES- EGFP-MMP was constructed by inserting the sense and antisense DNA of encoding the amino acid of the synthetic MMP enzyme cutting site into the eukaryotic expression vector pIRES-EGFE LAP and mTGF-β3 fragments were obtained by using RT-PCR and inserted into the upstream and downstream of MMP from pIRES-EGFP-MMP respectively, and the recombinant plasmid of pIRES-EGFP- LAP-MMP-mTGF-β3 was constructed, which was transferred to ADSCs. The ADSCs were cultured and divided in three groups： experimental group （MMP group）, negative control group （no MMP） and non-transfection group. The morphological changes were observed microscopically, and the expression of proteoglycan and type II collagen （Col II） was detected by using Alcian blue staining and immuno- histochemistry staining at 7th, 14th and 21st day after culture. The recombinant plasmid of pIRES-EGFP-LAP-MMP-mTGF-β3 was correctly constructed by methods of enzyme cutting and se- quencing analysis. The mTGF-β3 fusion protein was successfully expressed after transfection, and in the presence of the MMP, active protein mTGF-β3 was generated, which significantly promoted differ- entiation of ADSCs into chondrocytes and the expression of cartilage matrix. The novel fusion protein LAP-MMP-mTGF-β3 can targetedly induce differentiation of ADSCs into chondrocytes, which would open up prospects for target therapy of cartilage damage repair in future.
