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重组P128蛋白的表达纯化及其对抗牛源葡萄球菌活性研究
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作者 肖真真 汤明元 +2 位作者 张鑫宇 夏晓莉 孙怀昌 《中国兽医杂志》 CAS 北大核心 2018年第10期16-19,24,共5页
为了制备重组P128蛋白用于葡萄球菌性奶牛乳房炎治疗。将引入烟草蚀纹病毒蛋白酶(TEVp)识别位点的P128编码序列插入类弹性蛋白多肽(ELP)融合表达载体,转化大肠杆菌后用IPTG诱导融合蛋白表达,用温度敏感相变循环纯化,用TEVp活性包涵体切... 为了制备重组P128蛋白用于葡萄球菌性奶牛乳房炎治疗。将引入烟草蚀纹病毒蛋白酶(TEVp)识别位点的P128编码序列插入类弹性蛋白多肽(ELP)融合表达载体,转化大肠杆菌后用IPTG诱导融合蛋白表达,用温度敏感相变循环纯化,用TEVp活性包涵体切割融合标签,用最小抑菌浓度(MIC)和最小杀菌浓度(MBC)试验测定重组P128对奶牛乳房炎葡萄球菌抗菌活性。结果显示,融合蛋白以可溶性蛋白表达,两次相变循环纯化后的纯度达90%,产量为400 mg/mL; TEVp活性包涵体切割融合标签的效率接近100%,回收的重组P128纯度达98%,对牛源葡萄球菌的MIC和MBC分别为0. 47μg/mL和1. 88μg/mL;除个别多重耐药菌株外,重组P128蛋白对多数金黄色葡萄球菌和凝固酶阴性葡萄球菌的溶(杀)作用很强。这些研究结果表明,重组P128可用于葡萄球菌性奶牛乳房炎治疗。 展开更多
关键词 重组P128蛋白 融合表达 纯化 抗葡萄球菌活性
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蜚蠊肠道放线菌WA23‐4‐4的抗金黄色葡萄球菌次级代谢产物研究 被引量:1
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作者 叶宇歆 曾还雄 +4 位作者 陈柳汕 汪洁 刘文彬 金小宝 沈娟 《广东药科大学学报》 CAS 2021年第1期6-10,共5页
目的研究蜚蠊肠道内生放线菌WA23‐4‐4的抗金黄色葡萄球菌次级代谢产物。方法采用硅胶柱色谱、葡聚糖凝胶柱色谱、半制备高效液相等方法分离纯化菌株WA23‐4‐4次级代谢产物;采用紫外光谱(UV)、红外光谱(IR)、核磁共振光谱(NMR)、质谱(... 目的研究蜚蠊肠道内生放线菌WA23‐4‐4的抗金黄色葡萄球菌次级代谢产物。方法采用硅胶柱色谱、葡聚糖凝胶柱色谱、半制备高效液相等方法分离纯化菌株WA23‐4‐4次级代谢产物;采用紫外光谱(UV)、红外光谱(IR)、核磁共振光谱(NMR)、质谱(MS)等技术鉴定结构;微量肉汤稀释法体外测定抗金黄色葡萄球菌活性。结果从蜚蠊肠道内生放线菌WA23‐4‐4的乙酸乙酯萃取物中分离得到10个化合物:环(L‐脯‐L‐缬)二肽(1)、环(S‐脯‐S‐异亮)二肽(2)、环(R‐脯‐R‐亮)二肽(3)、环‐(L‐脯‐L‐酪)二肽(4)、环(甘‐亮)二肽(5)、环(丙‐酪)二肽(6)、反式对羟基肉桂酸(7)、邻氨基苯甲酸(8)、N‐[2‐(4‐羟苯基)‐乙基]乙酰胺(9)、环(丙‐缬)二肽(10)。其中化合物1、2、5对金黄色葡萄球菌(ATCC 25923)生长具有较强抑制作用(MIC值均为128μg/mL),化合物1、2对耐甲氧西林金黄色葡萄球菌(ATCC 25213)生长具有一定的抑制作用(MIC值均为256μg/mL)。结论化合物1、2、5可能是该放线菌的抗金黄色葡萄球菌活性成分。 展开更多
关键词 蜚蠊 放线菌 金黄色葡萄球菌活性
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林氏咽宁散体外抗金黄色葡萄球菌实验
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作者 仇翼 张敏 《食品与药品》 CAS 2006年第12A期46-48,共3页
目的探讨林氏咽宁散对金黄色葡萄球菌的体外抑菌作用。方法以琼脂平皿法、液体试管法观察其抗菌作用。结果对金黄色葡萄球菌有一定的抑菌作用。琼脂平皿法最低抑菌浓度为0.1g/mL;液体试管法最低抑菌浓度和最低杀菌浓度均为0.025g/mL。... 目的探讨林氏咽宁散对金黄色葡萄球菌的体外抑菌作用。方法以琼脂平皿法、液体试管法观察其抗菌作用。结果对金黄色葡萄球菌有一定的抑菌作用。琼脂平皿法最低抑菌浓度为0.1g/mL;液体试管法最低抑菌浓度和最低杀菌浓度均为0.025g/mL。结论林氏咽宁散体外抗菌活性测定显示对金黄色葡萄球菌有一定的抗菌作用。 展开更多
关键词 林氏咽宁散 金黄色葡萄球菌 活性
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嵌合蛋白P128的表达、纯化与抗牛源葡萄球菌活性研究 被引量:5
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作者 汤明元 刘文俊 +2 位作者 张鑫宇 夏晓莉 孙怀昌 《扬州大学学报(农业与生命科学版)》 CAS 北大核心 2016年第1期1-5,共5页
将抗葡萄球菌嵌合蛋白P128编码序列优化成大肠埃希菌偏爱密码子序列,插入脑膜炎奈瑟氏菌FrpC蛋白自加工元件(SPM)与类弹性蛋白多肽(ELP)融合表达载体,将重组载体pP128-SPM-ELP转化大肠埃希菌进行诱导表达,利用ELP的温度敏感可逆相变循环... 将抗葡萄球菌嵌合蛋白P128编码序列优化成大肠埃希菌偏爱密码子序列,插入脑膜炎奈瑟氏菌FrpC蛋白自加工元件(SPM)与类弹性蛋白多肽(ELP)融合表达载体,将重组载体pP128-SPM-ELP转化大肠埃希菌进行诱导表达,利用ELP的温度敏感可逆相变循环(ITC)进行融合蛋白纯化,利用SPM自切割活性进行融合蛋白切割和目的蛋白回收;以牛源金黄葡萄球菌为指示菌,对回收的嵌合蛋白P128进行溶(杀)菌活性测定。结果表明:P128-SPM-ELP融合蛋白获得高水平、可溶性表达,经过两轮ITC获得的融合蛋白纯度大于90%,其自切割效率接近100%,回收的嵌合蛋白P128对金黄葡萄球菌的最小抑菌浓度为8.4μg·mL-1,且有很强的杀菌作用。证明SPM-ELP融合表达与纯化系统是简单、经济、有效的重组蛋白表达与纯化系统,获得的嵌合蛋白P128可用于葡萄球菌性奶牛疾病防治。 展开更多
关键词 嵌合蛋白P128 融合表达 纯化 抗葡萄球菌活性
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广藿香内生真菌Fusarium falciforme R16次生代谢产物及其抗菌活性
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作者 袁怡悦 马青云 +2 位作者 谢晴宜 吴友根 赵友兴 《分子植物育种》 CAS 北大核心 2024年第12期4009-4017,共9页
广藿香是具有广泛应用价值的重要南药,为了探究其内生真菌的代谢属性及其抗菌作用,本研究采用组织培养法从广藿香植物花组织中分离和鉴定内生真菌,采用硅胶柱和半制备高效液相色谱等多种柱色谱分离方法对广藿香内生真菌Fusarium falcifo... 广藿香是具有广泛应用价值的重要南药,为了探究其内生真菌的代谢属性及其抗菌作用,本研究采用组织培养法从广藿香植物花组织中分离和鉴定内生真菌,采用硅胶柱和半制备高效液相色谱等多种柱色谱分离方法对广藿香内生真菌Fusarium falciforme R16的次生代谢产物进行分离纯化,运用NMR和MS等波谱方法鉴定化合物结构,对单体化合物进行抗菌活性测试。结果从内生真菌Fusarium falciforme R16发酵产物中分离得到12个化合物,分别鉴定为对羟基苯乙醇(1)、2-羟乙基-联二苯(2)、4-甲基-5,6-2H-吡喃-2-酮(3)、苯乙酸乙酯(4)、苯甲醚(5)、4-bromostilbene-O-methyl isopropionate(6)、2-甲基-1,4-苯二酚(7)、N-(4-羟苯乙基)-2-苯乙酰胺(8)、乙酸-4-戊烯酯(9)、2-(4-甲氧基苯)-2-丙醇(10)、4-methylideneoxane(11)和2-四氢萘酚(12)。抗菌活性结果显示化合物5,7和10对金黄色葡萄球菌和枯草芽孢杆菌具有一定抑制活性,化合物11和12对金黄色葡萄球菌具有弱的抑制活性,本研究结果为真菌抗菌活性物质发掘提供参考依据。 展开更多
关键词 广藿香 内生真菌 次级代谢产物 金黄色葡萄球菌活性 枯草芽孢杆菌活性
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重组人源β-防御素-2基因真核表达质粒转染293细胞的表达产物抗菌活性的研究 被引量:8
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作者 王慧 董碧蓉 周焱 《生物医学工程学杂志》 EI CAS CSCD 北大核心 2009年第2期371-373,共3页
利用分子生物学技术构建人源β-防御素-2(HBD-2)真核表达重组质粒pcDNA3.1-zeo(+)-HBD-2,经脂质体介导转染293细胞,RT-PCR检测到HBD-2基因成功转染293细胞。收集培养上清,采用肉汤对倍稀释法检测抗菌活性,结果显示培养上清对金黄色葡萄... 利用分子生物学技术构建人源β-防御素-2(HBD-2)真核表达重组质粒pcDNA3.1-zeo(+)-HBD-2,经脂质体介导转染293细胞,RT-PCR检测到HBD-2基因成功转染293细胞。收集培养上清,采用肉汤对倍稀释法检测抗菌活性,结果显示培养上清对金黄色葡萄球菌有抗菌活性,而在相同条件下未检测到抗铜绿假单胞菌活性,提示人源β-防御素-2(HBD-2)可能具有优势抗金黄色葡萄球菌活性。 展开更多
关键词 Β-防御素-2 金色葡萄球菌活性 293细胞
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Chitosan Extends the Shelf-life of Filleted Tilapia (Oreochromis niloticus) During Refrigerated Storage 被引量:9
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作者 CAO Rong LIU Qi +1 位作者 YIN Bangzhong WU Biao 《Journal of Ocean University of China》 SCIE CAS 2012年第3期408-412,共5页
Shelf-life extension of aquatic products is of significant economical importance. To determine the potential effect of chitosan on the shelf-life of filleted tilapia, this study analyzed the bacterial community divers... Shelf-life extension of aquatic products is of significant economical importance. To determine the potential effect of chitosan on the shelf-life of filleted tilapia, this study analyzed the bacterial community diversity in fresh and spoiled tilapia fillets stored at (4 ± 1)℃ and examined the antimicrobial activity of chitosan against relevant bacteria isolates. Results showed that Pseudomonas (20%) and Aeromonas (16%) were abundant in fresh tilapia fillets, whereas Pseudomonas (52%), Aeromonas (32%) and Staphylococcus (12%) were dominant in the spoiled samples. Chitosan showed wide-spectrum antibacterial activity against bacteria isolated from tilapia and 5.0 g L-1 chitosan was selected for application in preservation. We further determined the shelf-life of chitosan-treated, filleted tilapia stored at (4 ± 1)℃ based on microbiological, biochemical and sensory analyses. Results showed that the shelf-life of chitosan-treated, filleted tilapia was extended to 12 d, whereas that of untreated, control samples was 6 d. These indicate that chitosan, as a natural preservative, has great application potential in the shelf-life extension of tilapia fillets. 展开更多
关键词 Oreochromis niloticus bacterial flora chitosan refrigeration shelf-life
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Endophytic Fungi Isolated from Mangrove (Rhyzopora mucronata) and Its Antibacterial Activity on Staphylococcus aureus and Escherichia coli
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《Journal of Food Science and Engineering》 2011年第5期386-389,共4页
Disc diffusion method was used to investigate the antibacterial activity of methanol extract of five endophytic fungi isolated from mangrove (Rhizopora mucronata) on Staphylococcus aureus ATCC 9144 and Escherichia c... Disc diffusion method was used to investigate the antibacterial activity of methanol extract of five endophytic fungi isolated from mangrove (Rhizopora mucronata) on Staphylococcus aureus ATCC 9144 and Escherichia coli ATCC 8739, Fungi were isolated from leaf, stem and root of R. mucronata. Potato dextrose agar and potato dextrose broth were used for fungi purification and fermentation. Fungal metabolites were extracted with methanol (1 : 1 v/v) for 24 h before vacuum evaporated to yield the crude extracts. Antimicrobial activity test was done according to the Kirby-Bauer test. The inhibition zone around each disc was considered to measure antibacterial activity of endophytic fungi. Only three endophytic fungi showed inhibition activity on S. aureus ATCC 9144 and two endophytic fungi showed inhibition on E. coil ATCC 8739. The results suggested that the extracts have a high potential for enteric diseases treatment caused by foodbome pathogen. 展开更多
关键词 ANTIBACTERIAL endophytic fungi MANGROVE Rhyzopora mucronata.
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Effect of BEDGEN 40 on Growth, Anti-Oxidant Activity and Resistance against Aeromonas Hydrophila Infection in Tilapia (Oreochromis niloticus x O. aureus)
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作者 W. D. Zheng 《Feed & Livestock》 2012年第3期30-33,共4页
The globe artichoke has become important as a medicinal herb in recent years following the discovery of cynarin. This bitter-tasting compound, which is found in the leaves, improves liver and gall bladder function, st... The globe artichoke has become important as a medicinal herb in recent years following the discovery of cynarin. This bitter-tasting compound, which is found in the leaves, improves liver and gall bladder function, stimulates the secretion of digestive juices, especially bile, and lowers blood cholesterol levels. The leaves are anticholesterolemic, antirheumatic, cholagogue, digestive, diuretic, hypoglycaemic and lithontripic. BEDGEN 40 , a compound made with Cynara scolymus L. (artichoke) extract and choline chloride was evaluated for growth performance and anti-oxidant activity in tilapia. There were four treatment groups; group 1 was fed only with a basal diet and served as the control, group 2 - 4 were experimental groups that received 150, 300, or 600 mg/kg BEDGEN 40 , respectively, added to the basal diet and fed for 8 weeks. The results showed signifi- cant differences for weight gain, feed conversion, and protein efficiency in fish diets containing 150, 300, or 600 mg/kg BEDGEN 40 compared to the control. Generally, BEDGEN 40 improved the body condition factor and hepatosomatic index of tilapia, and reduced liver fat content. Fish consum- ing BEDGEN 40 also exhibited improved liver antioxidant status. BEDGEN 40 (300, or 600 mg/kg) was also found to significantly reduce the mortality of tilapia subsequently challenged with Aeromonas hydrophila. 展开更多
关键词 Tilapia (Oreochromis niloticus × O.aureus) BEDGEN 40 cynara extract cholinechloride growth performance anti-oxidant activity disease resistance
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放线菌Streptomyces sp. FJ3的核糖体工程改良与活性产物的分离 被引量:5
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作者 海乐 黄宇琪 +1 位作者 廖国建 胡昌华 《微生物学报》 CAS CSCD 北大核心 2011年第7期934-940,共7页
【目的】利用核糖体工程抗性筛选技术,获得有抗菌活性突变株,并对突变株新产生活性物质进行研究。【方法】以三峡库区筛选出的无抗菌活性放线菌野生株为出发菌,通过单菌落挑选与平板划线培养,分离筛选具有链霉素和利福平抗性突变株;通... 【目的】利用核糖体工程抗性筛选技术,获得有抗菌活性突变株,并对突变株新产生活性物质进行研究。【方法】以三峡库区筛选出的无抗菌活性放线菌野生株为出发菌,通过单菌落挑选与平板划线培养,分离筛选具有链霉素和利福平抗性突变株;通过摇瓶发酵和对发酵液进行纸片法活性测定,获得抗金葡菌活性突变株;采用高效液相色谱法(HPLC)分析其发酵液组分,通过LC-MS对变化峰进行分析;进行16S rDNA及形态学鉴定。【结果】链霉素和利福平对放线菌菌株FJ3的MIC分别为0.5μg/mL和110μg/mL;在FJ3突变菌株中,共获得24株链霉素突变菌株和20株利福平突变菌株,抗菌活性筛选显示6株具有抗菌活性,其中2株链霉素突变菌株对金葡菌有强抑菌活性,采用Doskochilova溶剂系统纸层析结果表明,该活性物质为一种核酸类抗生素,HPLC和LC-MS显示该活性物质可能为硫藤黄菌素。【结论】利用核糖体工程技术可以改变放线菌的次级代谢,获得具有生物活性的突变株,拓展药源放线菌活性菌株新资源。 展开更多
关键词 三峡库区 放线菌 核糖体工程 链霉素 金黄色葡萄球菌活性
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Synergistic antibacterial effect of honey and Herba Ocimi Basilici against some bacterial pathogens 被引量:4
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作者 Ali Talha Khalil Imran Khan +3 位作者 Kafeel Ahmad Yusra Ali Khan Momin Khan Muhammad Jaseem Khan 《Journal of Traditional Chinese Medicine》 SCIE CAS CSCD 2013年第6期810-814,共5页
OBJECTIVE: To evaluate the antibacterial activity of the combination of different honey brands and methanolic fraction of Herba Ocimi Basilici using agar well diffusion assay. METHODS: The antibacterial activity was... OBJECTIVE: To evaluate the antibacterial activity of the combination of different honey brands and methanolic fraction of Herba Ocimi Basilici using agar well diffusion assay. METHODS: The antibacterial activity was deter- mined against thirteen pathogenic bacterial clini- cal isolates including six gram negative (Klebsiella pneumonia, Pseudomonas aeroginosa, Escherichia coli, Salmonella typhi, Salmonella typhimirium, Xanthomonas campestris) and six gram positive strains (Enterococcus faecalis faecalis, Bacillus subti- lis, Staphylococcus aureus, Clostridium perfringens type C, Clostridium perfringens type D, CIostridium chauvoei). Agar well diffusion method was used while zones of inhibition were measured with verni- er scale. RESULTS: At higher concentration, all the honey brands showed good to significant activity. Thehighest activity was observed for Hamdard brand honey (27.60±0.40) against Enterococcus faecalis. CONCLUSION: These results revealed that combi- nations of plant extracts of Herba Ocimi Basilici with honey can be used for the development of potent and novel antibacterial agents. 展开更多
关键词 Anti-infective agents HONEY Gram-negative bacteria Gram-positive bacteria Well dif-fusion assay
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Enhanced antibacterial activity of silica nanorattles with ZnO combination nanoparticles against methicillin-resistant Staphylococcus aureus 被引量:1
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作者 Qianqian Chai Qiong wu +5 位作者 Tianlong Liu Longfei Tan Changhui Fu Xiangling Ren Yue Yang Xianwei Meng 《Science Bulletin》 SCIE EI CAS CSCD 2017年第17期1207-1215,共9页
Silica nanorattles(SNs) with zinc oxide(ZnO) combination nanoparticles are reported to inhibit methicillin-resistant Staphylococcus aureus(MRSA) for the first time. SNs loaded with ZnO nanoparticles,which can produce ... Silica nanorattles(SNs) with zinc oxide(ZnO) combination nanoparticles are reported to inhibit methicillin-resistant Staphylococcus aureus(MRSA) for the first time. SNs loaded with ZnO nanoparticles,which can produce free radicals, can cause severe damage to bacteria. ZnO nanoparticles not only provide free radicals in the combined nanostructures, which can inhibit the growth of bacteria, but also form nanorough surfaces with an irregular distribution of spikes on the SNs, which can enhance their adhesion to bacteria. Nanorough silica shell surfaces maintain the high activity and stability of small-sized ZnO nanoparticles and gather ZnO nanoparticles together to enhance production, which improves the efficiency of free radicals against the cytomembranes of bacterial cells. The enhanced adhesion of ZnO@SN nanoparticles to MRSA cells shortens the effective touching distance between free radicals and MRSA, which also improves antibacterial activity. As we expected, the ZnO@SN nanoparticles exhibit a better antibacterial effect than free ZnO nanoparticles against MRSA in vitro and in vivo. We also demonstrate that SNs loaded with ZnO nanoparticles can accelerate wound healing in MRSA skin inflammation models. This method of multilevel functionalization will be potentially applicable to the antibacterial field. 展开更多
关键词 Silica nanorattles ZnO nanoparticles Methicillin-resistant Staphylococcus aureus(MRSA) Rough surface Reactive oxygen species
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Biodegradable cationic e-poly-L-lysine-conjugated polymeric nanoparticles as a new effective antibacterial agent
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作者 赵瑞芳 王海 +3 位作者 季天骄 Greg Anderson 聂广军 赵宇亮 《Science Bulletin》 SCIE EI CAS CSCD 2015年第2期216-226,I0002,共12页
Biocompatible and biodegradable ε-poly-L- lysine (EPL)/poly (ε-caprolactone) (PCL) copolymer was designed and synthesized. The amphiphilic EPL-PCL copolymer could easily self-assembled into monodispersed nanop... Biocompatible and biodegradable ε-poly-L- lysine (EPL)/poly (ε-caprolactone) (PCL) copolymer was designed and synthesized. The amphiphilic EPL-PCL copolymer could easily self-assembled into monodispersed nanoparticles (NPs), which showed a broad-spectrum antibacterial activity against Escherichia coli, Staphylococcus aureus and Bacillus subtilis. Interestingly, the antibacterial efficacy of the novel NPs is more potent than the cationic peptide EPL. To explore the underlying mechanism of the biodegradable cationic NPs, various possible antibacterial pathways have been validated. The NPs have been found that they can disrupt bacterial walls/ membranes and induce the increasing in reactive oxygen species and alkaline phosphatase levels. More importantly, the self-assembled NPs induced the changes in bacterial osmotic pressure, resulting in cell invagination to form holes and cause the leakage of cytoplasm. Taken together, our results suggest that the EPL-PCL NPs can be further developed to be a promising antimicrobial agent to treat infectious diseases as surfactants and emulsifiers to enhance drug encapsulation efficiency and antimicrobial activity. 展开更多
关键词 EPL-PCL copolymers Biodegradable cationic nanoparticles Broad-spectrum antibacterial activity Disruption of bacterial walls/membranes Cell invagination
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