To establish a cell line for stable expression of human beta-defensin 3 (hBD3). Methods Full length cDNA of hBD3 was isolated from previously constructed pGEM-hBD3 and then inserted into pcDNA3. The recombinant vector...To establish a cell line for stable expression of human beta-defensin 3 (hBD3). Methods Full length cDNA of hBD3 was isolated from previously constructed pGEM-hBD3 and then inserted into pcDNA3. The recombinant vector identified carrying hBD3 with right direction was introduced into COS-7 cells by Lipofe-ctamine. Cell clones survived in G418-rich medium and with stable expression of hBD3 in both mRNA and protein levels were identified by RT-PCR and Western blot respectively. Genomic integration of the hBD3 gene with the COS-7 cells was confirmed by Southern dot blot and primary analysis. The antimicrobial activity of the secreted hBD3 was also evaluated. Results COS-7 cells transfected with pcDNA3-hBD3 expressed hBD3 stably in mRNA and protein level. Southern dot blot analysis showed successful integration of the hBD3 gene into the genome of COS-7 cell and the hBD-3 protein secreted into the culture medium showed antimicrobial activity. Conclusion We successfully established a hBD3-expressing cell line.展开更多
目的探究应用槐地凉血汤联合司库奇尤单抗治疗血热型白疕的疗效。方法收集应用司库奇尤单抗且经中医辨证为血热证的白疕患者,在基础治疗的同时治疗组予以司库奇尤单抗联合槐地凉血汤,对照组仅使用司库奇尤单抗,于基线、4周、8周、12周...目的探究应用槐地凉血汤联合司库奇尤单抗治疗血热型白疕的疗效。方法收集应用司库奇尤单抗且经中医辨证为血热证的白疕患者,在基础治疗的同时治疗组予以司库奇尤单抗联合槐地凉血汤,对照组仅使用司库奇尤单抗,于基线、4周、8周、12周时评估患者皮损面积及严重程度指数(psoriasis area and severity index,PASI)评分,皮肤病生活质量指数(dermatology life quality index,DLQI)评分,中医证候评分并比较基线、12周的人β防御素2(human beta defensin-2,HBD-2)与超敏C反应蛋白(hypersensitive C reactive protein,hs-CRP)值,进行统计分析。结果治疗组在PASI75、PASI90、DLQI 0/1应答率及中医证候评分改善方面优于对照组(P<0.05);两组在12周时较基线均能降低血清HBD-2值,且治疗组效果更明显(P<0.05);两组12周hs-CRP值均较基线有统计学意义(P<0.05)。结论应用司库奇尤单抗治疗血热型白疕的过程中联合槐地凉血汤可以更好消除皮损,改善生活质量,降低中医评分,缓解潜在系统炎症,且槐地凉血汤可能通过下调HBD-2参与控制白疕病程。展开更多
目的探索肺结核患者血清甲壳质酶蛋白40(chitinase protein 40,YKL-40)、β-防御素-2(human beta defensin 2,HBD-2)水平及其对肺结核的诊断价值。方法选取2021年6月—2023年6月山东省立医院收治的86例肺结核患者为肺结核组,根据痰涂片...目的探索肺结核患者血清甲壳质酶蛋白40(chitinase protein 40,YKL-40)、β-防御素-2(human beta defensin 2,HBD-2)水平及其对肺结核的诊断价值。方法选取2021年6月—2023年6月山东省立医院收治的86例肺结核患者为肺结核组,根据痰涂片结果分为活动性肺结核组(n=50)和非活动性肺结核组(n=36),另选取86例来自我院体检中心的健康受试者作为对照组。检测所有受试者血清中YKL-40、HBD-2水平并分析肺结核患者血清中YKL-40与HBD-2的相关性;采用受试者工作特征(receiver operating characteristic,ROC)曲线分析血清中YKL-40、HBD-2水平对肺结核及活动性肺结核的诊断价值。结果肺结核组血清YKL-40、HBD-2水平高于对照组(P均<0.05);活动性肺结核组血清YKL-40、HBD-2水平高于非活动性肺结核组(P均<0.05)。肺结核患者血清中YKL-40与HBD-2的水平呈正相关(r=0.601)。YKL-40、HBD-2及2者联合诊断肺结核的曲线下面积(area under the curve,AUC)分别为0.895、0.922、0.962,2者联合诊断优于单独诊断;YKL-40、HBD-2及2者联合诊断活动性肺结核的AUC分别为0.891、0.881、0.959,2者联合诊断优于单独诊断。结论肺结核患者血清YKL-40、HBD-2水平显著升高,且随病情加重而升高,2者对肺结核及活动性肺结核的具有一定的诊断价值。展开更多
β-防御素体外试验显示具广谱抗菌活性 ,本研究应用转基因方法评估其体内抗菌作用。构建大鼠 β-防御素 - 2 (r BD2 )重组质粒 p BK- CMV- r BD2和 p CDNA- 3.1- Myc- His(+) - r BD2 ,通过脂质体包裹的方法将重组质粒经气管滴入 ,检测...β-防御素体外试验显示具广谱抗菌活性 ,本研究应用转基因方法评估其体内抗菌作用。构建大鼠 β-防御素 - 2 (r BD2 )重组质粒 p BK- CMV- r BD2和 p CDNA- 3.1- Myc- His(+) - r BD2 ,通过脂质体包裹的方法将重组质粒经气管滴入 ,检测其在气管和肺组织表达情况 ,并应用肺组织匀浆上清菌落计数法检察对绿脓杆菌的肺清除率。结果显示在基因转染大鼠的气管和肺组织内均检测到 r BD2 - His融合蛋白基因 m RNA和蛋白的表达 ,说明脂质体包裹的重组 β-防御素 - 2 p BK- CMV- r BD2质粒可有效转染气道上皮组织。肺细菌清除率实验显示构建重组质粒 p BK-CMV- r BD2气道转染与对照相比可显著提高肺组织对绿脓杆菌的清除率 (n(8,p(0 .0 1)。本研究表明 β-防御素基因气道转染可提高呼吸道天然抗感染防御功能 ,可能在呼吸道感染的防治实践中具有潜在应用价值。展开更多
文摘To establish a cell line for stable expression of human beta-defensin 3 (hBD3). Methods Full length cDNA of hBD3 was isolated from previously constructed pGEM-hBD3 and then inserted into pcDNA3. The recombinant vector identified carrying hBD3 with right direction was introduced into COS-7 cells by Lipofe-ctamine. Cell clones survived in G418-rich medium and with stable expression of hBD3 in both mRNA and protein levels were identified by RT-PCR and Western blot respectively. Genomic integration of the hBD3 gene with the COS-7 cells was confirmed by Southern dot blot and primary analysis. The antimicrobial activity of the secreted hBD3 was also evaluated. Results COS-7 cells transfected with pcDNA3-hBD3 expressed hBD3 stably in mRNA and protein level. Southern dot blot analysis showed successful integration of the hBD3 gene into the genome of COS-7 cell and the hBD-3 protein secreted into the culture medium showed antimicrobial activity. Conclusion We successfully established a hBD3-expressing cell line.
文摘目的探究应用槐地凉血汤联合司库奇尤单抗治疗血热型白疕的疗效。方法收集应用司库奇尤单抗且经中医辨证为血热证的白疕患者,在基础治疗的同时治疗组予以司库奇尤单抗联合槐地凉血汤,对照组仅使用司库奇尤单抗,于基线、4周、8周、12周时评估患者皮损面积及严重程度指数(psoriasis area and severity index,PASI)评分,皮肤病生活质量指数(dermatology life quality index,DLQI)评分,中医证候评分并比较基线、12周的人β防御素2(human beta defensin-2,HBD-2)与超敏C反应蛋白(hypersensitive C reactive protein,hs-CRP)值,进行统计分析。结果治疗组在PASI75、PASI90、DLQI 0/1应答率及中医证候评分改善方面优于对照组(P<0.05);两组在12周时较基线均能降低血清HBD-2值,且治疗组效果更明显(P<0.05);两组12周hs-CRP值均较基线有统计学意义(P<0.05)。结论应用司库奇尤单抗治疗血热型白疕的过程中联合槐地凉血汤可以更好消除皮损,改善生活质量,降低中医评分,缓解潜在系统炎症,且槐地凉血汤可能通过下调HBD-2参与控制白疕病程。
文摘目的探索肺结核患者血清甲壳质酶蛋白40(chitinase protein 40,YKL-40)、β-防御素-2(human beta defensin 2,HBD-2)水平及其对肺结核的诊断价值。方法选取2021年6月—2023年6月山东省立医院收治的86例肺结核患者为肺结核组,根据痰涂片结果分为活动性肺结核组(n=50)和非活动性肺结核组(n=36),另选取86例来自我院体检中心的健康受试者作为对照组。检测所有受试者血清中YKL-40、HBD-2水平并分析肺结核患者血清中YKL-40与HBD-2的相关性;采用受试者工作特征(receiver operating characteristic,ROC)曲线分析血清中YKL-40、HBD-2水平对肺结核及活动性肺结核的诊断价值。结果肺结核组血清YKL-40、HBD-2水平高于对照组(P均<0.05);活动性肺结核组血清YKL-40、HBD-2水平高于非活动性肺结核组(P均<0.05)。肺结核患者血清中YKL-40与HBD-2的水平呈正相关(r=0.601)。YKL-40、HBD-2及2者联合诊断肺结核的曲线下面积(area under the curve,AUC)分别为0.895、0.922、0.962,2者联合诊断优于单独诊断;YKL-40、HBD-2及2者联合诊断活动性肺结核的AUC分别为0.891、0.881、0.959,2者联合诊断优于单独诊断。结论肺结核患者血清YKL-40、HBD-2水平显著升高,且随病情加重而升高,2者对肺结核及活动性肺结核的具有一定的诊断价值。
文摘β-防御素体外试验显示具广谱抗菌活性 ,本研究应用转基因方法评估其体内抗菌作用。构建大鼠 β-防御素 - 2 (r BD2 )重组质粒 p BK- CMV- r BD2和 p CDNA- 3.1- Myc- His(+) - r BD2 ,通过脂质体包裹的方法将重组质粒经气管滴入 ,检测其在气管和肺组织表达情况 ,并应用肺组织匀浆上清菌落计数法检察对绿脓杆菌的肺清除率。结果显示在基因转染大鼠的气管和肺组织内均检测到 r BD2 - His融合蛋白基因 m RNA和蛋白的表达 ,说明脂质体包裹的重组 β-防御素 - 2 p BK- CMV- r BD2质粒可有效转染气道上皮组织。肺细菌清除率实验显示构建重组质粒 p BK-CMV- r BD2气道转染与对照相比可显著提高肺组织对绿脓杆菌的清除率 (n(8,p(0 .0 1)。本研究表明 β-防御素基因气道转染可提高呼吸道天然抗感染防御功能 ,可能在呼吸道感染的防治实践中具有潜在应用价值。
