目的研究糖化血红蛋白变异指数(HGI)、尿微量白蛋白(u-ALB)及血清腱生蛋白C(TNC)与2型糖尿病视网膜病变(T2DR)的相关性。方法回顾性选择2021年4月至2023年5月于首都医科大学大兴教学医院接受治疗的2型糖尿病(T2DM)患者1390例的临床资料...目的研究糖化血红蛋白变异指数(HGI)、尿微量白蛋白(u-ALB)及血清腱生蛋白C(TNC)与2型糖尿病视网膜病变(T2DR)的相关性。方法回顾性选择2021年4月至2023年5月于首都医科大学大兴教学医院接受治疗的2型糖尿病(T2DM)患者1390例的临床资料,根据T2DR发生情况将其分为T2DR组(n=378)和非T2DR组(n=1012)。依据糖尿病视网膜病变早期治疗研究分类系统对T2DR患者进行分期,分为非增殖期T2DR组(n=275)和增殖期T2DR(n=103)。观察两组基线资料(性别、年龄、T2DM病程)、血糖[空腹血糖(FBG)、餐后2 h血糖(2 h PBG)]、血脂[高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)、总胆固醇、甘油三酯]、HGI、u-ALB及血清TNC水平;观察不同T2DR病变分期患者基线资料、血糖、血脂、HGI、u-ALB、血清TNC水平。采用Pearson相关分析对T2DM患者HGI、u-ALB、血清TNC与T2DR的相关性进行分析。采用多因素Logistic回归分析对影响T2DR发生的独立危险因素进行分析。结果T2DR组与非T2DR组的性别构成比、年龄比较,差异均无统计学意义(P>0.05);T2DR组T2DM病程、FPG、2 h PBG、HbA1c、HDL-C、LDL-C、总胆固醇、甘油三酯、HGI、u-ALB、TNC水平均大于非T2DR组,差异均有统计学意义(P<0.05)。增殖期T2DR组与非增殖期T2DR组的性别构成比、年龄比较,差异均无统计学意义(P>0.05);增殖期T2DR组的T2DM病程、FPG、2 h PBG、HbA1c、HDL-C、LDL-C、总胆固醇、甘油三酯、HGI、u-ALB、TNC水平均大于非增殖期T2DR组,差异均有统计学意义(P<0.05)。Pearson相关分析结果显示,HGI、u-ALB、TNC与T2DM患者发生T2DR呈正相关(P<0.05)。多因素Logistic回归分析结果显示,HbA1c、HGI、u-ALB及TNC为影响T2DM患者发生T2DR的独立危险因素。结论HGI、u-ALB、TNC的异常升高可促进T2DM患者T2DR的发生及进展,HbA1c、HGI、u-ALB及TNC为影响T2DM患者发生T2DR的独立危险因素。展开更多
目的:研究活化蛋白C(Activated protein C,APC)对大鼠皮瓣缺血再灌注损伤的作用及可能机制。方法:将80只SD雄性大鼠随机分为四组:对照组、药物对照组、模型组和治疗组,每组20只。观察术后72 h内模型大鼠皮瓣形态变化,HE染色观察大鼠皮...目的:研究活化蛋白C(Activated protein C,APC)对大鼠皮瓣缺血再灌注损伤的作用及可能机制。方法:将80只SD雄性大鼠随机分为四组:对照组、药物对照组、模型组和治疗组,每组20只。观察术后72 h内模型大鼠皮瓣形态变化,HE染色观察大鼠皮瓣组织病理学变化,TUNEL法染色观察皮瓣组织细胞凋亡,ELISA法检测血清中TNF-α、IL-6水平,用黄嘌呤氧化酶法和硫代巴比妥酸TBA比色法分别测定皮瓣组织中超氧化物歧化酶(SOD)活性、丙二醛(MDA)含量,Westernblot法检测皮瓣组织中Nrf-2、HO-1、γ-GCS蛋白表达水平。结果:与模型组比较,治疗组皮瓣红肿、坏死程度、病理损伤程度减弱;TUNEL法染色观察,与模型组比较,治疗组皮瓣组织细胞凋亡率减少(P<0.05);ELISA法检测发现,与模型组比较,治疗组血清中TNF-α、IL-6降低(P<0.05);与模型组比较,治疗组SOD活性升高(P<0.05),MDA含量降低(P<0.05);Westernblot法检测发现,与模型组比较,治疗组Nrf-2、HO-1、γ-GCS蛋白相对表达水平上升(P<0.05)。结论:APC能改善大鼠皮瓣缺血再灌注损伤,其机制可能与激活Nrf-2/HO-1信号通路,抑制氧化应激反应和减少细胞凋亡、炎症反应有关。展开更多
AIM:To evaluate the potential of two trabecular meshwork(TM)-specific promoters,Chitinase 3-like 1(Ch3L1)and matrix gla protein(MGP),for improving specificity and safety in glaucoma gene therapy based on self-compleme...AIM:To evaluate the potential of two trabecular meshwork(TM)-specific promoters,Chitinase 3-like 1(Ch3L1)and matrix gla protein(MGP),for improving specificity and safety in glaucoma gene therapy based on self-complementary AAV2(scAAV2)vector technologies.METHODS:An scAAV2 vector with C3 transferase(C3)as the reporter gene(scAAV2-C3)was selected.The scAAV2-C3 vectors were driven by Ch3L1(scAAV2-Ch3L1-C3),MGP(scAAV2-MGP-C3),enhanced MGP(scAAV2-eMGP-C3)and cytomegalovirus(scAAV2-CMV-C3),respectively.The cultured primary human TM cells were treated with each vector at different multiplicities of infections.Changes in cell morphology were observed by phase contrast microscopy.Actin stress fibers and Rho GTPases/Rho-associated protein kinase pathway-related molecules were assessed by immunofluorescence staining,real-time quantitative polymerase chain reaction and Western blot.Each vector was injected intracamerally into the one eye of each rat at low and high doses respectively.In vivo green fluorescence was visualized by a Micron III Retinal Imaging Microscope.Intraocular pressure(IOP)was monitored using a rebound tonometer.Ocular responses were evaluated by slit-lamp microscopy.Ocular histopathology analysis was examined by hematoxylin and eosin staining.RESULTS:In TM cell culture studies,the vectormediated C3 expression induced morphologic changes,disruption of actin cytoskeleton and reduction of fibronectin expression in TM cells by inhibiting the Rho GTPases/Rhoassociated protein kinase signaling pathway.At the same dose,these changes were significant in TM cells treated with scAAV2-CMV-C3 or scAAV2-Ch3L1-C3,but not in cells treated with scAAV2-eMGP-C3 or scAAV2-MGP-C3.At lowinjected dose,the IOP was significantly decreased in the scAAV2-Ch3L1-C3-injected eyes but not in scAAV2-MGPC3-injected and scAAV2-eMGP-C3-injected eyes.At highinjected dose,significant IOP reduction was observed in the scAAV2-eMGP-C3-injected eyes but not in scAAV2-MGP-C3-injected eyes.Similar to scAAV2-CMV-C3,scAAV2-Ch3L1-C3 vector showed efficient transduction both in the TM and corneal endothelium.In anterior segment tissues of scAAV2-eMGP-C3-injected eyes,no obvious morphological changes were found except for the TM.Inflammation was absent.CONCLUSION:In scAAV2-transduced TM cells,the promoter-driven efficiency of Ch3L1 is close to that of cytomegalovirus,but obviously higher than that of MGP.In the anterior chamber of rat eye,the transgene expression pattern of scAAV2 vector is presumably affected by MGP promoter,but not by Ch3L1 promoter.These findings would provide a useful reference for improvement of specificity and safety in glaucoma gene therapy using scAAV2 vector.展开更多
文摘目的研究糖化血红蛋白变异指数(HGI)、尿微量白蛋白(u-ALB)及血清腱生蛋白C(TNC)与2型糖尿病视网膜病变(T2DR)的相关性。方法回顾性选择2021年4月至2023年5月于首都医科大学大兴教学医院接受治疗的2型糖尿病(T2DM)患者1390例的临床资料,根据T2DR发生情况将其分为T2DR组(n=378)和非T2DR组(n=1012)。依据糖尿病视网膜病变早期治疗研究分类系统对T2DR患者进行分期,分为非增殖期T2DR组(n=275)和增殖期T2DR(n=103)。观察两组基线资料(性别、年龄、T2DM病程)、血糖[空腹血糖(FBG)、餐后2 h血糖(2 h PBG)]、血脂[高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)、总胆固醇、甘油三酯]、HGI、u-ALB及血清TNC水平;观察不同T2DR病变分期患者基线资料、血糖、血脂、HGI、u-ALB、血清TNC水平。采用Pearson相关分析对T2DM患者HGI、u-ALB、血清TNC与T2DR的相关性进行分析。采用多因素Logistic回归分析对影响T2DR发生的独立危险因素进行分析。结果T2DR组与非T2DR组的性别构成比、年龄比较,差异均无统计学意义(P>0.05);T2DR组T2DM病程、FPG、2 h PBG、HbA1c、HDL-C、LDL-C、总胆固醇、甘油三酯、HGI、u-ALB、TNC水平均大于非T2DR组,差异均有统计学意义(P<0.05)。增殖期T2DR组与非增殖期T2DR组的性别构成比、年龄比较,差异均无统计学意义(P>0.05);增殖期T2DR组的T2DM病程、FPG、2 h PBG、HbA1c、HDL-C、LDL-C、总胆固醇、甘油三酯、HGI、u-ALB、TNC水平均大于非增殖期T2DR组,差异均有统计学意义(P<0.05)。Pearson相关分析结果显示,HGI、u-ALB、TNC与T2DM患者发生T2DR呈正相关(P<0.05)。多因素Logistic回归分析结果显示,HbA1c、HGI、u-ALB及TNC为影响T2DM患者发生T2DR的独立危险因素。结论HGI、u-ALB、TNC的异常升高可促进T2DM患者T2DR的发生及进展,HbA1c、HGI、u-ALB及TNC为影响T2DM患者发生T2DR的独立危险因素。
文摘目的:研究活化蛋白C(Activated protein C,APC)对大鼠皮瓣缺血再灌注损伤的作用及可能机制。方法:将80只SD雄性大鼠随机分为四组:对照组、药物对照组、模型组和治疗组,每组20只。观察术后72 h内模型大鼠皮瓣形态变化,HE染色观察大鼠皮瓣组织病理学变化,TUNEL法染色观察皮瓣组织细胞凋亡,ELISA法检测血清中TNF-α、IL-6水平,用黄嘌呤氧化酶法和硫代巴比妥酸TBA比色法分别测定皮瓣组织中超氧化物歧化酶(SOD)活性、丙二醛(MDA)含量,Westernblot法检测皮瓣组织中Nrf-2、HO-1、γ-GCS蛋白表达水平。结果:与模型组比较,治疗组皮瓣红肿、坏死程度、病理损伤程度减弱;TUNEL法染色观察,与模型组比较,治疗组皮瓣组织细胞凋亡率减少(P<0.05);ELISA法检测发现,与模型组比较,治疗组血清中TNF-α、IL-6降低(P<0.05);与模型组比较,治疗组SOD活性升高(P<0.05),MDA含量降低(P<0.05);Westernblot法检测发现,与模型组比较,治疗组Nrf-2、HO-1、γ-GCS蛋白相对表达水平上升(P<0.05)。结论:APC能改善大鼠皮瓣缺血再灌注损伤,其机制可能与激活Nrf-2/HO-1信号通路,抑制氧化应激反应和减少细胞凋亡、炎症反应有关。
基金Supported by the National Natural Science Foundation of China(No.81900829,No.82070963)the Xiamen Medical and Health Guiding Project Fund Project(No.3502Z20214ZD1214)+1 种基金the Guangdong Basic and Applied Basic Research Foundation(No.2019A1515011234)the Science and Technology Innovation Committee of Shenzhen(No.JCYJ20210324125614039)。
文摘AIM:To evaluate the potential of two trabecular meshwork(TM)-specific promoters,Chitinase 3-like 1(Ch3L1)and matrix gla protein(MGP),for improving specificity and safety in glaucoma gene therapy based on self-complementary AAV2(scAAV2)vector technologies.METHODS:An scAAV2 vector with C3 transferase(C3)as the reporter gene(scAAV2-C3)was selected.The scAAV2-C3 vectors were driven by Ch3L1(scAAV2-Ch3L1-C3),MGP(scAAV2-MGP-C3),enhanced MGP(scAAV2-eMGP-C3)and cytomegalovirus(scAAV2-CMV-C3),respectively.The cultured primary human TM cells were treated with each vector at different multiplicities of infections.Changes in cell morphology were observed by phase contrast microscopy.Actin stress fibers and Rho GTPases/Rho-associated protein kinase pathway-related molecules were assessed by immunofluorescence staining,real-time quantitative polymerase chain reaction and Western blot.Each vector was injected intracamerally into the one eye of each rat at low and high doses respectively.In vivo green fluorescence was visualized by a Micron III Retinal Imaging Microscope.Intraocular pressure(IOP)was monitored using a rebound tonometer.Ocular responses were evaluated by slit-lamp microscopy.Ocular histopathology analysis was examined by hematoxylin and eosin staining.RESULTS:In TM cell culture studies,the vectormediated C3 expression induced morphologic changes,disruption of actin cytoskeleton and reduction of fibronectin expression in TM cells by inhibiting the Rho GTPases/Rhoassociated protein kinase signaling pathway.At the same dose,these changes were significant in TM cells treated with scAAV2-CMV-C3 or scAAV2-Ch3L1-C3,but not in cells treated with scAAV2-eMGP-C3 or scAAV2-MGP-C3.At lowinjected dose,the IOP was significantly decreased in the scAAV2-Ch3L1-C3-injected eyes but not in scAAV2-MGPC3-injected and scAAV2-eMGP-C3-injected eyes.At highinjected dose,significant IOP reduction was observed in the scAAV2-eMGP-C3-injected eyes but not in scAAV2-MGP-C3-injected eyes.Similar to scAAV2-CMV-C3,scAAV2-Ch3L1-C3 vector showed efficient transduction both in the TM and corneal endothelium.In anterior segment tissues of scAAV2-eMGP-C3-injected eyes,no obvious morphological changes were found except for the TM.Inflammation was absent.CONCLUSION:In scAAV2-transduced TM cells,the promoter-driven efficiency of Ch3L1 is close to that of cytomegalovirus,but obviously higher than that of MGP.In the anterior chamber of rat eye,the transgene expression pattern of scAAV2 vector is presumably affected by MGP promoter,but not by Ch3L1 promoter.These findings would provide a useful reference for improvement of specificity and safety in glaucoma gene therapy using scAAV2 vector.
