A new method was used to preparing genomic DNA from Microbacterium sp.quickly and efficiently.DNA quantity and purity was measured by UV absorbance.Integrity of the genomic DNA was tested by agarose gel eletrophoresis...A new method was used to preparing genomic DNA from Microbacterium sp.quickly and efficiently.DNA quantity and purity was measured by UV absorbance.Integrity of the genomic DNA was tested by agarose gel eletrophoresis.The DNA prepared by this method was sufficiently pure for PCR.This method saves time and cost,practices easily as well.展开更多
The carbendazim（MBC） hydrolyzing enzyme gene was cloned and heterologously expressed in Escherichia coli BL21（DE3） from a newly isolated MBC-degrading bacterium strain Microbacterium sp. strain djl-6F. High perfor...The carbendazim（MBC） hydrolyzing enzyme gene was cloned and heterologously expressed in Escherichia coli BL21（DE3） from a newly isolated MBC-degrading bacterium strain Microbacterium sp. strain djl-6F. High performance liquid chromatography-mass spectrometry（HPLC-MS）analysis revealed that purified MheI-6F protein catalyzes direct hydrolysis of MBC into2-aminobenzimidazole（2-AB） with a high turnover rate and moderate affinity（Kmof6.69 μmol/L and kcatof 160.88/min） without the need for any cofactors. The optimal catalytic condition of MheI-6F was identified as 45°C, pH 7.0. The enzymatic activity of MheI-6F was found to be diminished by metal ions, and strongly inhibited by sodium dodecyl sulfate（SDS）.Through generating amino acid mutations in MheI-6F, Cys16 and Cys222 were identified as the catalytic groups that are essential for the hydrolysis of MBC. This is the first report on the biodegradation of MBC at the enzymatice level.展开更多
目的探讨血清表面活性蛋白D(surfactant protein D,SP-D)、黏蛋白1(mucin 1,MUC1)、锌指蛋白A20(zinc finger protein A20,A20)水平对输血相关性急性肺损伤(transfusion-related acute lung injury,TRALI)患者临床预后的预测价值,以期...目的探讨血清表面活性蛋白D(surfactant protein D,SP-D)、黏蛋白1(mucin 1,MUC1)、锌指蛋白A20(zinc finger protein A20,A20)水平对输血相关性急性肺损伤(transfusion-related acute lung injury,TRALI)患者临床预后的预测价值,以期临床个体化干预提供参考。方法选取2020年3月—2025年3月河北省胸科医院收治的TRALI患者249例为研究对象,根据输血后30 d内预后情况,分别纳入预后良好组(178例)、预后不良组(71例)。比较2组一般临床资料及血清SP-D、MUC1、A20水平,Logistic回归分析血清SP-D、MUC1、A20水平对TRALI患者预后不良的影响因素,受试者工作特征(receiver operating characteristic,ROC)曲线分析血清SP-D、MUC1、A20水平单独及联合检测对预后不良的预测效能,并采用一致性分析进行外部验证。结果2组输血次数、发血至输血时间、过敏史、急性生理与慢性健康评分系统Ⅱ(acute physiology and chronic health evaluationⅡ,APACHEⅡ)评分比较,差异有统计学意义(P<0.05);预后不良组血清SP-D、MUC1、A20水平分别为(89.54±21.36)g/L、(22.97±5.14)kU/L、(14.53±1.96)mg/L,明显高于预后良好组的(78.61±18.05)g/L、(16.28±4.32)kU/L、(12.67±1.84)mg/L,差异有统计学意义(P<0.05);Logistic回归分析显示,输血次数、发血至输血时间、过敏史均是TRALI患者预后不良的影响因素(P<0.05),APACHEⅡ评分及血清SP-D、MUC1、A20均与TRALI患者预后不良显著相关(P<0.05);ROC分析显示,血清SP-D、MUC1、A20联合预测预后不良的曲线下面积(area under the curve,AUC)为0.904(95%CI:0.860~0.938),优于各指标单独预测价值(Z=2.507、3.016、3.042,均P<0.05),且外部验证显示,联合预测预后不良与临床实际的符合率为95.00%,Kappa值为0.870(95%CI:0.617~0.982)差异有统计学意义(P<0.05)。结论血清SP-D、MUC1、A20均是TRALI患者预后不良的独立影响因素,联合检测对预后不良具有较高预测价值,可作为TRALI患者预后的潜在预测因子,并可指导临床工作。展开更多
文摘A new method was used to preparing genomic DNA from Microbacterium sp.quickly and efficiently.DNA quantity and purity was measured by UV absorbance.Integrity of the genomic DNA was tested by agarose gel eletrophoresis.The DNA prepared by this method was sufficiently pure for PCR.This method saves time and cost,practices easily as well.
基金supported by the Hi-Tech Research and Development Program(863) of China(No.2012AA101404)the National Natural Science Foundation of China(No.31301700)+1 种基金the Science and Technology Program of BaoJi(No.2013R6-3)the Key Subject Project of Baoji University of Arts and Sciences(No.ZK0919)
文摘The carbendazim（MBC） hydrolyzing enzyme gene was cloned and heterologously expressed in Escherichia coli BL21（DE3） from a newly isolated MBC-degrading bacterium strain Microbacterium sp. strain djl-6F. High performance liquid chromatography-mass spectrometry（HPLC-MS）analysis revealed that purified MheI-6F protein catalyzes direct hydrolysis of MBC into2-aminobenzimidazole（2-AB） with a high turnover rate and moderate affinity（Kmof6.69 μmol/L and kcatof 160.88/min） without the need for any cofactors. The optimal catalytic condition of MheI-6F was identified as 45°C, pH 7.0. The enzymatic activity of MheI-6F was found to be diminished by metal ions, and strongly inhibited by sodium dodecyl sulfate（SDS）.Through generating amino acid mutations in MheI-6F, Cys16 and Cys222 were identified as the catalytic groups that are essential for the hydrolysis of MBC. This is the first report on the biodegradation of MBC at the enzymatice level.
文摘目的探讨血清表面活性蛋白D(surfactant protein D,SP-D)、黏蛋白1(mucin 1,MUC1)、锌指蛋白A20(zinc finger protein A20,A20)水平对输血相关性急性肺损伤(transfusion-related acute lung injury,TRALI)患者临床预后的预测价值,以期临床个体化干预提供参考。方法选取2020年3月—2025年3月河北省胸科医院收治的TRALI患者249例为研究对象,根据输血后30 d内预后情况,分别纳入预后良好组(178例)、预后不良组(71例)。比较2组一般临床资料及血清SP-D、MUC1、A20水平,Logistic回归分析血清SP-D、MUC1、A20水平对TRALI患者预后不良的影响因素,受试者工作特征(receiver operating characteristic,ROC)曲线分析血清SP-D、MUC1、A20水平单独及联合检测对预后不良的预测效能,并采用一致性分析进行外部验证。结果2组输血次数、发血至输血时间、过敏史、急性生理与慢性健康评分系统Ⅱ(acute physiology and chronic health evaluationⅡ,APACHEⅡ)评分比较,差异有统计学意义(P<0.05);预后不良组血清SP-D、MUC1、A20水平分别为(89.54±21.36)g/L、(22.97±5.14)kU/L、(14.53±1.96)mg/L,明显高于预后良好组的(78.61±18.05)g/L、(16.28±4.32)kU/L、(12.67±1.84)mg/L,差异有统计学意义(P<0.05);Logistic回归分析显示,输血次数、发血至输血时间、过敏史均是TRALI患者预后不良的影响因素(P<0.05),APACHEⅡ评分及血清SP-D、MUC1、A20均与TRALI患者预后不良显著相关(P<0.05);ROC分析显示,血清SP-D、MUC1、A20联合预测预后不良的曲线下面积(area under the curve,AUC)为0.904(95%CI:0.860~0.938),优于各指标单独预测价值(Z=2.507、3.016、3.042,均P<0.05),且外部验证显示,联合预测预后不良与临床实际的符合率为95.00%,Kappa值为0.870(95%CI:0.617~0.982)差异有统计学意义(P<0.05)。结论血清SP-D、MUC1、A20均是TRALI患者预后不良的独立影响因素,联合检测对预后不良具有较高预测价值,可作为TRALI患者预后的潜在预测因子,并可指导临床工作。
