AIM： To study the effects of IκBα and its mutants （IκBαM, IκBα3N, IκBαM44C） on NF-κB, p53 and their downstream target genes. The relationship of NF-κB, p53, and IκBα was further discussed. METHODS： pEC...AIM： To study the effects of IκBα and its mutants （IκBαM, IκBα3N, IκBαM44C） on NF-κB, p53 and their downstream target genes. The relationship of NF-κB, p53, and IκBα was further discussed. METHODS： pECFP-IκBα, pECFP-IκBαM （amino acides 1-317, Ser32, 36A）, pECFP-IκBα243N （amino acides 1-243）, pECFP-IκBα244C （amino acides 24±317）, pEYFP-p65 and pp53-DsRed were constructed and transfected to ASTC-α-1 cells. Cells were transfected with pECFP-Cl as a control. 30 h after the transfection, location patterns of NF-κB, p53 and IκBα（IκBαM, IκBα243N, IκBα224C） were observed by a laser scanning microscope （LSM510/ConfoCor2, Zeiss）. RNA extraction and reverse transcription were performed in cells transfected or co-transfected with different plasmids. Effects of IκBα and its mutants on the transprition level of NF-κB, NF-κB downstream target gene TNF-α, p53 and p53 downstream target gene Bax were observed by real time QT-PCR. In all experiments β-actin was reference. Results are expressed as the target/reference ratio of the sample divided by the target/reference ratio of the control. Different transfected cells were incubated with CCK-8 for 2 h in the incubator. Then the absorbance at 450 nm was measured by using a microplate reader. RESULTS： Cells that were transfected with p53- DsRed revealed a predominant nuclear localization. YFP-p65 mainly existed in the cytoplasm. Cells were transfected with CFP-IκBα, CFP-IκBαM, and CFP-IκBα243N respectively and revealed a predominant cytosolic localization. However, cells transfected of CFP-IκBα244C revealed a predominant nuclear localization. The rnRNA levels of p65, TNF-α, p53 and Bax in CFP-IκBα transfected cells did not change significantly, while in YFP-p65/CFP-IκBα co-transfected cells, IκBα decreased the transcription of p65 downstream gene TNF-α （2.24 ± 0.503） compared with the YFP-p65/ CFP-C1 co-transfected cells （5.08 ± 0.891） （P 〈 0.05）. Phosphorylation defective IκBα （IκBαM） decreased the transcription levels of all the four genes compared with the control （P 〈 0.05）. The N terminus of IκBα（IκBα243N） increased the transcription of NF-κB （1.84 ± 0.176） and TNF-α （1.51 ± 0.203） a little bit. However, the C terminus of IκBα（IκBα244C） increased the transcription of NF-κB, TNF-α, p53 and Bax significantly （8.29 ± 1.662, 14.16 ± 2.121, 10.2 ± 0.621, 3.72 ± 0.346） （P 〈 0.05）. The CCK-8 experiment also showed that IκBα244C and p53 synergistically mediate apoptosis. CONCLUSIONS： IκBα and its mutants （IκBαM, IκBα243N, IκBαM244C） have different effects on NF- KB and p53 signaling pathways, according to their different structures. IκBαbounds with NF-KB and p53 in cytoplasm steadily, and inhibits both of the two signaling pathways, p53 and IκBα244C may be co-factor in inducing apoptosis. The C terminal of IκBαnhanced cell death, which suggests that it may be a pro-apoptotic protein existed in cells.展开更多
OBJECTIVE:To evaluate the diagnostic efficacy of Qisexingtai hand diagnostic method in the diagnosis of coronary artery disease(CAD).METHODS:This was an investigator-initiated,prospective,multi-center,cross-sectional ...OBJECTIVE:To evaluate the diagnostic efficacy of Qisexingtai hand diagnostic method in the diagnosis of coronary artery disease(CAD).METHODS:This was an investigator-initiated,prospective,multi-center,cross-sectional study.All the participants from three hospitals in China had been diagnosed by both Qisexingtai hand diagnostic method and coronary angiography.We compared the two diagnostic methods to calculate the sensitivity,the specificity,the omission diagnostic rate,the mistaken diagnostic rate and accuracy in order to evaluate the diagnostic efficacy of Qisexingtai hand diagnostic method for CAD.RESULTS:A total of 326 subjects were enrolled,diagnosed by both Qisexingtai hand diagnostic method and coronary angiography.As a result,there were 166 positive cases according to Qisexingtai hand diagnostic method,and 131 positive cases according to coronary angiography.Compared with the results of coronary angiography,the sensitivity of Qisexingtai hand diagnostic method was 80.2%,the specificity was 68.7%,the omission diagnostic rate was 19.8%,the mistaken diagnostic rate was 31.3%,and the accuracy was 73.3%.Area under the receiver operating characteristic curve was estimated as 0.735 for all,and 0.718,0.735,0.783 for the three sub-centers.CONCLUSION:Qisexingtai hand diagnostic method with high accuracy and sensitivity has certain application value in the diagnosis of CAD.展开更多
目的:探讨儿童早期预警评分(Pediatric Early Warning Score,PEWS)用于病毒性脑炎患儿病情判断的有效性。方法:选取2018年7月至2019年7月在邢台市人民医院住院期间因病情加重需要转入儿科重症监护病房(pediatric intensive care unit,PI...目的:探讨儿童早期预警评分(Pediatric Early Warning Score,PEWS)用于病毒性脑炎患儿病情判断的有效性。方法:选取2018年7月至2019年7月在邢台市人民医院住院期间因病情加重需要转入儿科重症监护病房(pediatric intensive care unit,PICU)治疗的28例病毒性脑炎患儿,设为PICU组;选取不需要转入PICU治疗的35例轻症患者,作为轻症组。记录轻症组入院时的PEWS和PICU组患儿转入PICU12 h前的PEWS评分。比较两组PEWS评分的差异,并进行受试者工作特征(receiver operatingcharacters,ROC)曲线分析。结果:两组患儿PEWS评分中位数(四分位数间距)分别是轻症组3分(2~4分),PICU组6分(5~7分),两组间评分差异具有统计学意义(χ2=163.836,P<0.01)。患儿需转入PICU的PEWS评分为5分或单项得分≥3分。结论:改良PEWS评分可用于病毒性脑炎患儿病情严重程度判断和转入PICU的预测。展开更多
基金the National Natural Science Foundation of China No. 60378043 and 30470494)the Natural ScienceFoundation of Guangdong Province No. 015012 and 04010394
文摘AIM： To study the effects of IκBα and its mutants （IκBαM, IκBα3N, IκBαM44C） on NF-κB, p53 and their downstream target genes. The relationship of NF-κB, p53, and IκBα was further discussed. METHODS： pECFP-IκBα, pECFP-IκBαM （amino acides 1-317, Ser32, 36A）, pECFP-IκBα243N （amino acides 1-243）, pECFP-IκBα244C （amino acides 24±317）, pEYFP-p65 and pp53-DsRed were constructed and transfected to ASTC-α-1 cells. Cells were transfected with pECFP-Cl as a control. 30 h after the transfection, location patterns of NF-κB, p53 and IκBα（IκBαM, IκBα243N, IκBα224C） were observed by a laser scanning microscope （LSM510/ConfoCor2, Zeiss）. RNA extraction and reverse transcription were performed in cells transfected or co-transfected with different plasmids. Effects of IκBα and its mutants on the transprition level of NF-κB, NF-κB downstream target gene TNF-α, p53 and p53 downstream target gene Bax were observed by real time QT-PCR. In all experiments β-actin was reference. Results are expressed as the target/reference ratio of the sample divided by the target/reference ratio of the control. Different transfected cells were incubated with CCK-8 for 2 h in the incubator. Then the absorbance at 450 nm was measured by using a microplate reader. RESULTS： Cells that were transfected with p53- DsRed revealed a predominant nuclear localization. YFP-p65 mainly existed in the cytoplasm. Cells were transfected with CFP-IκBα, CFP-IκBαM, and CFP-IκBα243N respectively and revealed a predominant cytosolic localization. However, cells transfected of CFP-IκBα244C revealed a predominant nuclear localization. The rnRNA levels of p65, TNF-α, p53 and Bax in CFP-IκBα transfected cells did not change significantly, while in YFP-p65/CFP-IκBα co-transfected cells, IκBα decreased the transcription of p65 downstream gene TNF-α （2.24 ± 0.503） compared with the YFP-p65/ CFP-C1 co-transfected cells （5.08 ± 0.891） （P 〈 0.05）. Phosphorylation defective IκBα （IκBαM） decreased the transcription levels of all the four genes compared with the control （P 〈 0.05）. The N terminus of IκBα（IκBα243N） increased the transcription of NF-κB （1.84 ± 0.176） and TNF-α （1.51 ± 0.203） a little bit. However, the C terminus of IκBα（IκBα244C） increased the transcription of NF-κB, TNF-α, p53 and Bax significantly （8.29 ± 1.662, 14.16 ± 2.121, 10.2 ± 0.621, 3.72 ± 0.346） （P 〈 0.05）. The CCK-8 experiment also showed that IκBα244C and p53 synergistically mediate apoptosis. CONCLUSIONS： IκBα and its mutants （IκBαM, IκBα243N, IκBαM244C） have different effects on NF- KB and p53 signaling pathways, according to their different structures. IκBαbounds with NF-KB and p53 in cytoplasm steadily, and inhibits both of the two signaling pathways, p53 and IκBα244C may be co-factor in inducing apoptosis. The C terminal of IκBαnhanced cell death, which suggests that it may be a pro-apoptotic protein existed in cells.
基金Supported by National Key R&D Program of China-Study on Operating Principles of Screening,Evaluation,Promotion and Application for Diagnosis and Treatment Technology of Characteristic Folk Traditional Chinese Medicine(No.2019YFC1708400)。
文摘OBJECTIVE:To evaluate the diagnostic efficacy of Qisexingtai hand diagnostic method in the diagnosis of coronary artery disease(CAD).METHODS:This was an investigator-initiated,prospective,multi-center,cross-sectional study.All the participants from three hospitals in China had been diagnosed by both Qisexingtai hand diagnostic method and coronary angiography.We compared the two diagnostic methods to calculate the sensitivity,the specificity,the omission diagnostic rate,the mistaken diagnostic rate and accuracy in order to evaluate the diagnostic efficacy of Qisexingtai hand diagnostic method for CAD.RESULTS:A total of 326 subjects were enrolled,diagnosed by both Qisexingtai hand diagnostic method and coronary angiography.As a result,there were 166 positive cases according to Qisexingtai hand diagnostic method,and 131 positive cases according to coronary angiography.Compared with the results of coronary angiography,the sensitivity of Qisexingtai hand diagnostic method was 80.2%,the specificity was 68.7%,the omission diagnostic rate was 19.8%,the mistaken diagnostic rate was 31.3%,and the accuracy was 73.3%.Area under the receiver operating characteristic curve was estimated as 0.735 for all,and 0.718,0.735,0.783 for the three sub-centers.CONCLUSION:Qisexingtai hand diagnostic method with high accuracy and sensitivity has certain application value in the diagnosis of CAD.
文摘目的:探讨儿童早期预警评分(Pediatric Early Warning Score,PEWS)用于病毒性脑炎患儿病情判断的有效性。方法:选取2018年7月至2019年7月在邢台市人民医院住院期间因病情加重需要转入儿科重症监护病房(pediatric intensive care unit,PICU)治疗的28例病毒性脑炎患儿,设为PICU组;选取不需要转入PICU治疗的35例轻症患者,作为轻症组。记录轻症组入院时的PEWS和PICU组患儿转入PICU12 h前的PEWS评分。比较两组PEWS评分的差异,并进行受试者工作特征(receiver operatingcharacters,ROC)曲线分析。结果:两组患儿PEWS评分中位数(四分位数间距)分别是轻症组3分(2~4分),PICU组6分(5~7分),两组间评分差异具有统计学意义(χ2=163.836,P<0.01)。患儿需转入PICU的PEWS评分为5分或单项得分≥3分。结论:改良PEWS评分可用于病毒性脑炎患儿病情严重程度判断和转入PICU的预测。
