目的探讨2型糖尿病(T2DM)患者线粒体呼吸链复合酶Ⅰ、Ⅱ、Ⅲ活性与疲劳之间的潜在关系。方法选取2022年1月1日—2022年12月31日于泗洪县第一人民医院就诊的130例T2DM患者作为研究对象,根据疲劳量表评分分为糖尿病无疲乏组和糖尿病疲乏...目的探讨2型糖尿病(T2DM)患者线粒体呼吸链复合酶Ⅰ、Ⅱ、Ⅲ活性与疲劳之间的潜在关系。方法选取2022年1月1日—2022年12月31日于泗洪县第一人民医院就诊的130例T2DM患者作为研究对象,根据疲劳量表评分分为糖尿病无疲乏组和糖尿病疲乏组。采集患者的血样,进行常规的空腹血糖(FBG)、糖化血红蛋白(HbA1c)检测,分别使用辅酶Q10(CoQ10)酶联免疫试剂盒和线粒体呼吸链复合物活性定量检测试剂盒检测CoQ10和呼吸链复合酶Ⅰ、Ⅱ、Ⅲ的活性。采用独立样本t检验、双相关Pearson检验、ROC曲线、二元logistic回归分析评估各项临床指标与糖尿病疲劳风险的相关性。结果2组患者FBG、HbA1c、24 h平均血糖(24 h MBG)及24 h血糖标准差(24 h SDBG)和简单自测抑郁量表(PHQ-9)、广泛性焦虑量表(GAD-7)评分差异无统计学意义(P>0.05)。疲乏组患者CoQ10和线粒体呼吸链复合酶Ⅰ、Ⅱ、Ⅲ水平均低于无疲乏组患者(P<0.05)。T2DM患者CoQ10水平和线粒体呼吸链复合酶Ⅰ、Ⅱ、Ⅲ活性与疲劳量表评分呈负相关(r=-0.451~-0.906,P<0.05)。性别(女性)、年龄(60~69岁/70~79岁)、病程(10~年)均为影响T2DM患者产生疲劳感的独立危险因素。结论线粒体呼吸链复合酶Ⅰ、Ⅱ、Ⅲ活性降低与T2DM患者疲劳感产生密切相关,而且对T2DM患者疲劳感的产生具有预测价值。展开更多
目的构建人类MHC-Ⅰ类链相关基因A(MICA)的真核表达载体,转染人舌鳞癌脑高转移Tca8113-Tb细胞,建立稳定过表达MICA基因的口腔鳞癌细胞系。方法采用PCR技术扩增pCMV-SPORT6-MICA中编码MICA基因的cDNA序列,重组至有绿色荧光蛋白标记的真...目的构建人类MHC-Ⅰ类链相关基因A(MICA)的真核表达载体,转染人舌鳞癌脑高转移Tca8113-Tb细胞,建立稳定过表达MICA基因的口腔鳞癌细胞系。方法采用PCR技术扩增pCMV-SPORT6-MICA中编码MICA基因的cDNA序列,重组至有绿色荧光蛋白标记的真核表达载体pEGFP-N1,构建最终的表达载体pEGFP-N1-MICA,脂质体法转染Tca8113-Tb细胞,G418筛选,荧光显微镜下观察绿色荧光蛋白的表达,有限稀释法建立稳定过表达MICA基因的Tca8113-Tb细胞系,RT-PCR、real time PCR和免疫细胞化学检测MICA在该细胞中的表达。结果通过PCR技术获取了MICA基因并成功克隆入载体,测序鉴定该序列与GenBank中的序列相同。转染的细胞可见绿色荧光蛋白表达,RT-PCR、real time PCR及免疫细胞化学检测到目的基因MICA在转染细胞中为过表达。结论 pEGFP-N1-MICA真核表达载体的成功构建与稳定转染Tca8113-Tb细胞系的建立,为进一步研究该基因的功能奠定了良好的实验基础。展开更多
Background In addition to the well-known antibodies against human leukocyte antigens （HLA）-induced kidney-graft rejection, polymorphic major-histocompatibility-complex （MHC） class Ⅰ-related chain A （MICA） antig...Background In addition to the well-known antibodies against human leukocyte antigens （HLA）-induced kidney-graft rejection, polymorphic major-histocompatibility-complex （MHC） class Ⅰ-related chain A （MICA） antigens can elicit antibodies and have been suggested to play a role in the antibody-mediated allograft rejection （AMR）. We carded out a prospective study of MICA antibodies in post-renal transplant patients to determine the association between MICA antibodies, C4d staining, histological features, and graft outcome.Methods We tested 52 patients who had biopsy results due to graft dysfunction. The MICA antibodies in concurrent sera were determined by Luminex. All patients were followed up for one year after renal biopsy. The influence of antibody production on the function of graft was analyzed.Results Antibodies against MICA were positive in 15 out of the 52 patients （28.9%）. The presence of MICA antibodies was associated with renal-allograft deterioration. During one-year follow-up, the estimated glomerular filtration rate （eGFR） decreased （24.0±3.4）% among recipients with anti-MICA antibodies. However, among recipients without anti-MICA antibodies, the eGFR has declined only （8.4＋3.0）% （P=0.017）. The association between C4d staining,histological features and MICA antibody production was found no significant difference.Conclusion Besides anti-HLA antibodies, the presence of post-transplant MICA antibody is associated with poor graft outcome and increases the risk of graft failure.展开更多
Animals living in extremely high elevations have to adapt to low temperatures and low oxygen availability(hypoxia),but the underlying genetic mechanisms associated with these adaptations are still unclear.The mitochon...Animals living in extremely high elevations have to adapt to low temperatures and low oxygen availability(hypoxia),but the underlying genetic mechanisms associated with these adaptations are still unclear.The mitochondrial respiratory chain can provide>95%of the ATP in animal cells,and its efficiency is influenced by temperature and oxygen availability.Therefore,the respiratory chain complexes(RCCs)could be important molecular targets for positive selection associated with respiratory adaptation in high-altitude environments.Here,we investigated positive selection in 5 RCCs and their assembly factors by analyzing sequences of 106 genes obtained through RNA-seq of all 15 Chinese Phrynocephalus lizard species,which are distributed from lowlands to the Tibetan plateau(average elevation>4,500 m).Our results indicate that evidence of positive selec-tion on RCC genes is not significantly different from assembly factors,and we found no difference in selective pressures among the 5 complexes.We specifically looked for positive selection in line-ages where changes in habitat elevation happened.The group of lineages evolving from low to high altitude show stronger signals of positive selection than lineages evolving from high to low elevations.Lineages evolving from low to high elevation also have more shared codons under positive selection,though the changes are not equivalent at the amino acid level.This study advan-ces our understanding of the genetic basis of animal respiratory metabolism evolution in extreme high environments and provides candidate genes for further confirmation with functional analyses.展开更多
文摘目的探讨2型糖尿病(T2DM)患者线粒体呼吸链复合酶Ⅰ、Ⅱ、Ⅲ活性与疲劳之间的潜在关系。方法选取2022年1月1日—2022年12月31日于泗洪县第一人民医院就诊的130例T2DM患者作为研究对象,根据疲劳量表评分分为糖尿病无疲乏组和糖尿病疲乏组。采集患者的血样,进行常规的空腹血糖(FBG)、糖化血红蛋白(HbA1c)检测,分别使用辅酶Q10(CoQ10)酶联免疫试剂盒和线粒体呼吸链复合物活性定量检测试剂盒检测CoQ10和呼吸链复合酶Ⅰ、Ⅱ、Ⅲ的活性。采用独立样本t检验、双相关Pearson检验、ROC曲线、二元logistic回归分析评估各项临床指标与糖尿病疲劳风险的相关性。结果2组患者FBG、HbA1c、24 h平均血糖(24 h MBG)及24 h血糖标准差(24 h SDBG)和简单自测抑郁量表(PHQ-9)、广泛性焦虑量表(GAD-7)评分差异无统计学意义(P>0.05)。疲乏组患者CoQ10和线粒体呼吸链复合酶Ⅰ、Ⅱ、Ⅲ水平均低于无疲乏组患者(P<0.05)。T2DM患者CoQ10水平和线粒体呼吸链复合酶Ⅰ、Ⅱ、Ⅲ活性与疲劳量表评分呈负相关(r=-0.451~-0.906,P<0.05)。性别(女性)、年龄(60~69岁/70~79岁)、病程(10~年)均为影响T2DM患者产生疲劳感的独立危险因素。结论线粒体呼吸链复合酶Ⅰ、Ⅱ、Ⅲ活性降低与T2DM患者疲劳感产生密切相关,而且对T2DM患者疲劳感的产生具有预测价值。
文摘目的构建人类MHC-Ⅰ类链相关基因A(MICA)的真核表达载体,转染人舌鳞癌脑高转移Tca8113-Tb细胞,建立稳定过表达MICA基因的口腔鳞癌细胞系。方法采用PCR技术扩增pCMV-SPORT6-MICA中编码MICA基因的cDNA序列,重组至有绿色荧光蛋白标记的真核表达载体pEGFP-N1,构建最终的表达载体pEGFP-N1-MICA,脂质体法转染Tca8113-Tb细胞,G418筛选,荧光显微镜下观察绿色荧光蛋白的表达,有限稀释法建立稳定过表达MICA基因的Tca8113-Tb细胞系,RT-PCR、real time PCR和免疫细胞化学检测MICA在该细胞中的表达。结果通过PCR技术获取了MICA基因并成功克隆入载体,测序鉴定该序列与GenBank中的序列相同。转染的细胞可见绿色荧光蛋白表达,RT-PCR、real time PCR及免疫细胞化学检测到目的基因MICA在转染细胞中为过表达。结论 pEGFP-N1-MICA真核表达载体的成功构建与稳定转染Tca8113-Tb细胞系的建立,为进一步研究该基因的功能奠定了良好的实验基础。
文摘Background In addition to the well-known antibodies against human leukocyte antigens （HLA）-induced kidney-graft rejection, polymorphic major-histocompatibility-complex （MHC） class Ⅰ-related chain A （MICA） antigens can elicit antibodies and have been suggested to play a role in the antibody-mediated allograft rejection （AMR）. We carded out a prospective study of MICA antibodies in post-renal transplant patients to determine the association between MICA antibodies, C4d staining, histological features, and graft outcome.Methods We tested 52 patients who had biopsy results due to graft dysfunction. The MICA antibodies in concurrent sera were determined by Luminex. All patients were followed up for one year after renal biopsy. The influence of antibody production on the function of graft was analyzed.Results Antibodies against MICA were positive in 15 out of the 52 patients （28.9%）. The presence of MICA antibodies was associated with renal-allograft deterioration. During one-year follow-up, the estimated glomerular filtration rate （eGFR） decreased （24.0±3.4）% among recipients with anti-MICA antibodies. However, among recipients without anti-MICA antibodies, the eGFR has declined only （8.4＋3.0）% （P=0.017）. The association between C4d staining,histological features and MICA antibody production was found no significant difference.Conclusion Besides anti-HLA antibodies, the presence of post-transplant MICA antibody is associated with poor graft outcome and increases the risk of graft failure.
基金This work was supported by the National Natural Science Foundation of China(Grant No.31772447)the Second Tihetan Platean Scientife Expedition and Research Program(STEP),Grant No.2019 QZKK0501.VS.did this research within the framework of the HSE University Basic Research Program.
文摘Animals living in extremely high elevations have to adapt to low temperatures and low oxygen availability(hypoxia),but the underlying genetic mechanisms associated with these adaptations are still unclear.The mitochondrial respiratory chain can provide>95%of the ATP in animal cells,and its efficiency is influenced by temperature and oxygen availability.Therefore,the respiratory chain complexes(RCCs)could be important molecular targets for positive selection associated with respiratory adaptation in high-altitude environments.Here,we investigated positive selection in 5 RCCs and their assembly factors by analyzing sequences of 106 genes obtained through RNA-seq of all 15 Chinese Phrynocephalus lizard species,which are distributed from lowlands to the Tibetan plateau(average elevation>4,500 m).Our results indicate that evidence of positive selec-tion on RCC genes is not significantly different from assembly factors,and we found no difference in selective pressures among the 5 complexes.We specifically looked for positive selection in line-ages where changes in habitat elevation happened.The group of lineages evolving from low to high altitude show stronger signals of positive selection than lineages evolving from high to low elevations.Lineages evolving from low to high elevation also have more shared codons under positive selection,though the changes are not equivalent at the amino acid level.This study advan-ces our understanding of the genetic basis of animal respiratory metabolism evolution in extreme high environments and provides candidate genes for further confirmation with functional analyses.
