CN116370406B - 一种用于治疗痔疮纳米酶可注射水凝胶 - Google Patents
一种用于治疗痔疮纳米酶可注射水凝胶 Download PDFInfo
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Abstract
本发明提供了一种用于治疗痔疮纳米酶可注射水凝胶,本发明用微波法制备了铁掺杂磷钼杂多酸纳米材料(Fe‑PMs),将具有抗氧化及抗炎特性石榴皮水提物及单宁酸(TA)作为还原剂制备银纳米,具有抗菌壳聚糖及聚乙烯醇为凝胶分子骨架,利用Fe‑PMs的超高纳米酶模拟氧化酶活性,氧化含丰富酚羟基的TA,维持苯酚‑醌的动态氧化还原平衡,制备纳米酶可注射水凝胶,Fe‑MoSe2产生具有杀菌能力的活性氧,AgNPs及凝胶基底抗菌作用,达到高效抗菌效果;石榴皮水提物具有的抗氧化及收敛作用,TA的强抗炎作用,制备的纳米酶可注射水凝胶用于修复及改善内痔、外痔、混合痔及肛裂、肛瘘手术后引起的出血、疼痛、肛门坠胀等症状。
Description
技术领域
本发明涉及纳米材料抗菌技术领域,具体为一种用于治疗痔疮纳米酶可注射水凝胶。
背景技术
痔疮是一种常见、多发病,影响世界4%的人口。痔疮患者经常因为无痛出血、脱垂、血栓形成引起的疼痛或瘙痒就诊,开始多采用保守治疗,包括提高饮食中的纤维素类、 增加水的摄入量及局部用药,保守治疗失败的患者可采用套扎、红外线凝结及硬化剂注射疗法。对于三、四度痔疮及采用其他方法未见好转的急性期痔疮,外科手术治疗是首选, 痔切除术被认为痔疮手术的金标准,但其具有术后疼痛、出血、水肿的诟病。水凝胶因其能提供湿润的环境、吸收伤口渗出物、抗细菌等特性,一直被认为是很有前途的伤口敷料。目前市场上用于痔疮治疗凝胶包括壳聚糖凝胶、纳米银抗菌水凝胶等,但由于性能相对单一,对于痔疮发病期、术后恢复等治疗效果相当有限。
石榴皮中大量存在的植物多酚使石榴皮提取物(PPE)及所含单宁酸(TA)具有抗炎、抗氧化、抗菌等多种生物活性;壳聚糖具有良好的生物相容性、生物降解性、低毒性、无抗原性、抗微生物活性、凝血和止血作用的优点成为伤口敷料重要的基材。纳米酶因能产生活性氧而被广泛用作新型抗菌剂,目前研究最多的是用其过氧化化物酶,可以催化过氧化氢(H2O2)生成羟基自由基(·OH),达到抗菌目的,但用到有毒的H2O2,而氧化物酶不使用有毒的H2O2,更具有应用价值。
多金属氧酸盐是一类具有高负电荷、独特螯合性质和多样结构的金属-氧化物阴离子,它们还具有独特的电子转移性质和调控纳米材料电荷组成的能力。因此,推测多金属氧酸盐可以作为构筑基元,在不需要额外表面修饰的情况下,探索富含电荷的纳米酶。基于此,本发明通过微波法合成了铁掺杂磷钼杂多酸纳米材料(Fe-PMs),令人惊讶的是,得到的Fe-PMs表现出优异的类氧化物酶活性,Fe/Mo双金属氧化还原电对促进了活性Fe2+的再生,两者协同促进了Fe-PMs的催化增强。
发明内容
本发明提供了一种用于治疗痔疮纳米酶可注射水凝胶,本发明用微波法制备了铁掺杂磷钼杂多酸纳米材料(Fe-PMs),将具有抗氧化及抗炎特性石榴皮提取物及所含单宁酸(TA)作为还原剂、制备银纳米(AgNPs),具有抗菌壳聚糖(CS)及聚乙烯醇(PVA)为凝胶分子骨架,利用Fe-PMs的超高纳米酶模拟氧化酶(OXD)活性,氧化含丰富酚羟基的TA,维持苯酚-醌的动态氧化还原平衡,制备纳米酶可注射水凝胶,Fe-PMs纳米酶表现出诱导水凝胶自固化的高催化活性,为水凝胶提供了长期和可重复的粘附性,类似于贻贝的粘附。利用Fe-MoSe2产生具有杀菌能力的活性氧(ROS),AgNPs及凝胶基底抗菌作用,达到高效抗菌效果;同时利用石榴皮提取物具有的抗氧化及收敛作用,TA 的强抗炎作用,制备的纳米酶可注射水凝胶用于修复及改善内痔、外痔、混合痔及肛裂、肛瘘手术后引起的出血、疼痛、肛门坠胀等症状,促进痔核缩小、防止痔核脱垂和减轻痔黏膜充血水肿等症状,亦可用于痔疮等肛门疾病治疗的抗菌护理,能够对痔疮的伤口起到有效的愈合作用。AgNPs/Fe-PMs可注射水凝胶具有止痒、低毒、安全,渗透性好等特点,具有促进组织修复与再生的作用。在注入肛门时更为方便,只要将凝胶轻轻挤压就可以注入肛门,减少对创口的碰触和按压,也减少了注入药物的时间和复杂程度。
本发明制备用于治疗痔疮纳米酶可注射水凝胶,按以下步骤进行:
(1)铁掺杂磷钼杂多酸纳米材料(Fe-PMs)制备:0.15 - 0.20 g磷钼酸(PM)溶于15-20 mL纯水中,加入0.25 - 0.30 g氯化铁,常温搅拌25-30 min后,转移到微波消解罐中,140 ℃持续消解3-4 h,冷却至室温,离心,经0.22μm滤膜过滤,真空干燥,即得Fe-PMs;
(2)AgNPs/Fe-PMs可注射水凝胶制备:于90℃水浴加热搅拌溶解配制PVA溶液,冷到至40-45°C,加入AgNO3溶液,搅拌30-60 min,冷却至室温,作为A液;将Fe-PMs与石榴皮提取物水溶液按体积比1:10混合搅拌,作为B液;将4-5重量份A液和1重量份B液混合,常温避光搅拌30-40min,得C液;用1%的醋酸配制壳聚糖(CS)溶液,取CS溶液和C液按4:1的体积比混合搅拌10-20min,即得AgNPs/Fe-PMs可注射水凝胶。其中,石榴皮提取物浓度为2-3mg/mL,PVA浓度为10-12%(w/v),AgNO3溶液浓度为2-3mg/mL, CS浓度为2-3%(w/v),Fe-PMs浓度为0.1-0.5mg/mL。
所述的石榴皮提取物按以下方法制备:将干燥石榴皮粉碎,过40-60目筛,称取石榴皮粉末0.1-0.2g,加入50-60mL 40%-50%乙醇,在50℃条件下超声25-30min,离心,取上清液,烘箱60-70℃烘1-2h后,得石榴皮提取物,根据需求用纯水配制为所需浓度。
所述的离心是在4000-6000r/min处理10-15min。
AgNPs/Fe-PMs可注射水凝胶具有强模拟氧化酶(OXD)活性及模拟超氧歧化酶(SOD)活性,模拟OXD活性在弱酸(pH 4-5)条件下活性最强,而模拟SOD活性在中性或弱碱性(pH 7-8)条件下活性最强。
AgNPs/Fe-PMs可注射水凝胶,将用于修复及改善内痔、外痔、混合痔及肛裂、肛瘘手术后引起的出血、疼痛、肛门坠胀等症状。
本发明的优点在于:
1、本发明制备的铁掺杂磷钼杂多酸纳米材料(Fe-PMs),具有超强模拟氧化酶(OXD)及超氧歧化酶(SOD)特性,利用具有抗氧化及抗炎特性石榴皮提取物及所含单宁酸作为还原剂制备银纳米(AgNPs),利用抗菌壳聚糖(CS)及聚乙烯醇(PVA)为凝胶分子骨架,Fe-PMs具有的超高纳米酶模拟氧化酶(OXD)活性,氧化含丰富酚羟基的TA,维持苯酚-醌的动态氧化还原平衡,制备纳米酶可注射水凝胶,Fe-PMs纳米酶表现出诱导水凝胶自固化的高催化活性,为水凝胶提供了长期和可重复的粘附性,类似于贻贝的粘附;
2、本发明将天然产物活性物质与纳米材料有效结合,利用其抗菌、收敛、消炎等功效制备纳米酶可注射水凝胶,用于痔疮治疗,包括修复及改善内痔、外痔、混合痔及肛裂、肛瘘手术后引起的出血、疼痛、肛门坠胀等症状,促进痔核缩小、防止痔核脱垂和减轻痔黏膜充血水肿等症状,亦可用于痔疮等肛门疾病治疗的抗菌护理,能够对痔疮的伤口起到有效的愈合作用;
3、AgNPs/Fe-PMs可注射水凝胶具有止痒、低毒、安全,渗透性好等特点,具有促进组织修复与再生的作用;在注入肛门时更为方便,只要将凝胶轻轻挤压就可以注入肛门,减少对创口的碰触和按压,也减少了注入药物的时间和复杂程度。
附图说明
图1为实施例1中Fe-PMs的TEM图;
图2为实施例1中AgNPs/Fe-PMs可注射水凝胶的TEM图;
图3为实施例1中Fe-PMs及AgNPs/Fe-PMs可注射水凝胶的Zeta电位图;
图4为实施例1中AgNPs/Fe-PMs可注射水凝胶氧化TMB(拟OXD)的紫外可见吸收光谱图,图中(1)为单独TMB,(2)TMB和AgNPs/Fe-PMs可注射水凝胶;
图5为实施例1 AgNPs/Fe-PMs可注射水凝胶还原硝基蓝四氮唑(NBT)的抑制率,清除能力O2 ·-的能力(拟SOD)的紫外可见吸收光谱图,图中(1)为单独NBT,(2)NBT和AgNPs/Fe-PMs可注射水凝胶;
图6为实施例1中AgNPs/Fe-PMs可注射水凝胶的黏性;
图7为实施例1 AgNPs/Fe-PMs可注射水凝胶细胞毒性实验结果;
图8为实施例1 AgNPs/Fe-PMs可注射水凝胶抗菌实验结果,图中空白为不加可注射水凝胶;
图9 为实施例1各组小鼠肛缘水肿及结肠变化图;
图10为实施例1各组小鼠中性炎症细胞因子用药后的变化图;
图11 为实施例1 小鼠模型肛缘组织病理学染色图。
实施方式
下面将结合具体的实施例对本发明的技术方案作进一步详细地描述说明,但本发明的保护范围并不仅限于此。
实施例1:铁掺杂磷钼杂多酸纳米材料(Fe-PMs)制备及性能
(1)铁掺杂磷钼杂多酸纳米材料(Fe-PMs)制备:0.15 g磷钼酸(PM)溶于15mL纯水中,加入0.25 g氯化铁,常温搅拌30 min后,转移到微波消解罐中,140 ℃持续消解4 h,冷却至室温,离心,经0.22μm滤膜过滤,60℃真空干燥24h,即得Fe-PMs;
(2)石榴皮提取物制备:将干燥石榴皮粉碎,过50目筛,称取石榴皮粉末0.1g,加入50 mL 50%乙醇,在50℃条件下超声25 min,4000 r/min处理15min,取上清液,烘箱70℃烘2h后,得石榴皮提取物,取100mg石榴皮提取物溶于10mL去离子水中,制备10mg/mL石榴皮提取物;
(3)AgNPs/Fe-PMs可注射水凝胶制备:于90℃水浴加热搅拌溶解配制12%的PVA溶液,冷到至45°C,加入浓度为2.5 mg/mL,搅拌40 min,冷却至室温,作为A液;将0.2 mg/mLFe-PMs与2 mg/mL石榴皮提取物水溶液按体积比1:10混合搅拌,作为B液;将5重量份A液和1重量份B液混合,常温避光搅拌35 min,得C液;用1%的醋酸配制3%的壳聚糖(CS)溶液,取CS溶液和C液按4:1的体积比混合搅拌10 min,即得AgNPs/Fe-PMs可注射水凝胶;
(4)纳米材料的TEM及Zeta电位测试:将合成的Fe-PMs及AgNPs/Fe-PMs可注射水凝胶冷冻干燥后,进行透射电镜及扫描电镜(TEM及SEM)及Zeta电位测试(图1-3),从TEM图(图1)看出,Fe-PMs呈现良好的分散性带平均尺寸为2.7±0.7 nm的准球形形貌,晶格空间为0.22 nm,对应于石墨的(100)晶面; AgNPs/Fe-PMs可注射水凝胶SEM图呈现高度多孔结构(图2);Fe-PMs的Zeta电位为负,而AgNPs/Fe-PMs可注射水凝胶为正(图3),因为CS的存在使得材料表面带正电荷,从而有利于与带负电荷的细胞膜结合,改变细胞膜通透性,有利于抗菌;
(5)采用TMB显色反应测定纳米酶的拟氧化酶(OXD)活性:将100µg/mL AgNPs/Fe-PMs可注射水凝胶5μL、100mmol/L的TMB 50µL,加入到pH5.0醋酸盐缓冲溶液2mL中,混合均匀,静置10min,用紫外-可见分光光度计在654 nm处测量吸光度,每个样品测量3次,取平均值,吸收光谱图见图4,由图可知,AgNPs/Fe-PMs可注射水凝胶有相当高的拟氧化物酶活性;
(6)采用超氧物歧化酶(SOD)对氮蓝四唑(NBT)光还原的抑制作用来确定其酶活性大小。在有氧化物质存在下,核黄素可被光还原,被还原的核黄素在有氧条件下极易再氧化而产生·O2 -,·O2 -可将NBT还原为蓝色的甲腙,并且在560 nm 处有最大吸收;而SOD可清除·O2 -,从而抑制了甲腙的形成;将100µg/mL AgNPs/Fe-PMs可注射水凝胶100μL与含500μL2.25 mM NBT、500μL 60 μM核黄素、500μL 14.5mM甲硫氨酸溶液和500μL 3mM EDTA-Na2溶液的pH=7.8磷酸盐缓冲液1.9 mL混匀在LED灯光下反应30 min,空白不加AgNPs/Fe-PMs可注射水凝胶。用紫外-可见分光光度计在580 nm处测定吸光度,每个样品测量3次,取平均值,结果如图5;从图5中可以看出,AgNPs/Fe-PMs可注射水凝胶表现出相当高的拟SOD活性;
(7)AgNPs/Fe-PMs可注射水凝胶黏性:AgNPs/Fe-PMs可注射水凝胶具有自黏性,可反复黏附于各种表面;如图6(a,b)所示,水凝胶可以粘附在各种表面,包括樱桃番茄、玻璃瓶、小鼠肝脏和金属坚果;定量拉伸-粘附测试结果表明水凝胶对猪皮、玻璃瓶、聚乙烯材质离心管、香蕉、柠檬、西红柿和橘子等物体的粘附强度分别为65、45、44、51、32、48和46 kPa;
(8)细胞毒性测试:采用CCK-8细胞活力试剂盒检测纳米酶的细胞毒性,具体实验,将人脐静脉内皮细胞(HUVECs, 北纳创联生物科技有限公司)接种于96孔板中培养24h后,分别用不同浓度的AgNPs/Fe-PMs可注射水凝胶(0、5、10、20、40、80、160 μg/mL,以Ag+含量计算)分别孵育12、24、36h,分次用PBS漂洗细胞,每孔加入CCK-8溶液至10%浓度,37℃孵育,在450nm处测定吸光度;CCK-8分析(图7)显示AgNPs/Fe-PMs可注射水凝胶对细胞均无明显毒性;
(9)AgNPs/Fe-PMs可注射水凝胶抗菌试验
以下菌种从北纳创联生物科技有限公司获得;
实验方法:以耐甲氧西林金黄色葡萄球菌(MRSA, ATCC 43300)和大肠杆菌(E. coli,ATCC 25922)为代表革兰氏阴性菌株。采用平板计数法,通过计数CFU数来判定AgNPs/Fe-PMs可注射水凝胶的抗菌性能。首先,将上述菌种在固体Luria-Bertani(LB)培养基和固体营养肉汤培养基孵育24 h,用接种环挑取少量形成的菌落,接种到对应液体培养基(5mL)中,然后,在37℃、180rpm恒温摇床下震荡孵育12h后即可获得细菌悬浮液(1×108 CFU/mL),用无菌磷酸盐缓冲液(PBS)稀释到1×105 CFU/mL。材料分为四组:空白对照组,Fe-PMs可注射水凝胶、AgNPs可注射水凝胶及AgNPs/Fe-PMs可注射水凝胶组。将培养的细菌加入磷酸盐缓冲液中作为空白对照组,其他组与不同凝胶敷料混合,凝胶敷料浓度为50μg/mL,在37℃下将孵育60min,菌悬液经过稀释后(100 μL)均匀涂布在LB固体培养基和营养肉汤固体培养基上,在37℃下培养24 h,计算菌落数,判断抗菌性能;
结果如图8所示,空白对照组几乎没有抗菌性能,Fe-PMs可注射水凝胶、AgNPs可注射水凝胶及AgNPs/Fe-PMs可注射水凝胶组,MRSA和E. coli的抑菌率分别约为33.4%、52.0%及 99.9%,36.1%、64.7%及 100.0%,AgNPs/Fe-PMs可注射水凝胶组对MRSA和E. coli有近100%的杀菌率;
(10)小鼠痔疮模型构建及痔疮愈合试验
小鼠痔疮模型:所有动物实验均符合《动物护理指导原则》。选取10周龄、体重18~20g的雄性ICR小鼠作为实验动物。在雄性 ICR小鼠肛门内0.5cm 处推注0.05 mL 20%乙酸溶液,24 h后,具有肛周肿胀、白色溃烂面现象的小鼠即为造模成功小鼠,总共40只。随机分为5组(每组 8只小鼠):空白组、Fe-PMs 凝胶组、AgNPs凝胶组、AgNPs/Fe-PMs可注射水凝胶及马应龙麝香痔疮膏阳性对照组,空白注射无菌PBS,其余凝胶浓度为50μg/mL,将上述凝胶(各300μL)注入小鼠肛门,每日1次,连续5d;
小鼠肛缘水肿及结肠变化评估:治疗 5d 后,空白组肛缘明显肿胀,Fe-PMs 凝胶组、AgNPs凝胶组肛缘边缘高于皮肤,马应龙麝香痔疮膏肛缘可见,而AgNPs/Fe-PMs可注射水凝胶完全无水肿;同时通过小鼠结肠变化评估痔疮愈合情况,结果表明,痔疮模型组小鼠结肠出现黄色肿胀,结合小鼠肛门红肿、便秘情况,应是难以代谢的排泄物;经过凝胶治疗后,结肠长度有所恢复,肠内容物逐渐呈现健康黑色状态,AgNPs/Fe-PMs可注射水凝胶最为明显,与肛缘水肿结果一致,见图9;
ELISA 检测IL-1β、IL-6、TNF-α水平:末次给药后,小鼠禁食不禁水12 h后,分别取小鼠肛缘组织外周血。采用ELISA法检测各组小鼠血浆中IL-1β、IL-6、TNF-α;结果表明,治疗 5d 后,AgNPs/Fe-PMs凝胶组肛缘组织中炎性细胞因子IL-1β、IL-6、TNF-α水平显著低于其他凝胶组及空白组(P<0. 05),见图10;
肛缘组织HE 染色和免疫荧光:末次给药24 h后,取小鼠肛缘组织。采用10%甲醛溶液固定组织24 h,然后将固定的组织脱水,石蜡包埋,制备石蜡切片。将制备的石蜡切片进行HE染色;取石蜡切片,脱蜡、固定,山羊血清进行封闭,然后分别使用抗 Ly6G 抗体和抗F4/80 抗体对组织中中性粒细胞和巨噬细胞浸润情况进行染色,洗片后再使用二抗室温孵育1 h;治疗5d 后,病理染色结果显示,治疗组肛缘组织中炎性细胞浸润显著少于其他凝胶组及空白组(P<0. 05),见图11。
以上结果表明,本发明制备的纳米酶AgNPs/Fe-PMs可注射水凝胶有二重酶活性,通过纳米酶模拟OXD活性起到抗菌、石榴皮提取物抗氧化、收敛作用及纳米酶模拟SOD活性清除自由起到的消炎作用,对小鼠痔疮起到有效的治愈作用。
Claims (2)
1.一种用于治疗痔疮纳米酶可注射水凝胶,其特征在于,制备步骤如下:
(1)铁掺杂磷钼杂多酸纳米材料Fe-PMs的制备:0.15 - 0.20 g磷钼酸PM溶于15-20 mL纯水中,加入0.25 - 0.30 g氯化铁,常温搅拌25-30 min后,转移到微波消解罐中,140 ℃持续消解3-4 h,冷却至室温,在4000- 6000r/min处理10-15min离心,经0.22μm滤膜过滤,真空干燥,即得Fe-PMs;
(2)AgNPs/Fe-PMs可注射水凝胶的制备:于90℃水浴加热搅拌溶解配制PVA溶液,冷到至40-45°C,加入AgNO3溶液,搅拌30-60 min,冷却至室温,作为A液;将Fe-PMs与石榴皮提取物水溶液按体积比1:10混合搅拌,作为B液;将4-5重量份A液和1重量份B液混合,常温避光搅拌30-40min,得C液;用1%的醋酸配制壳聚糖CS溶液,取CS溶液和C液按4:1的体积比混合搅拌10-20min,即得AgNPs/Fe-PMs可注射水凝胶;其中,石榴皮提取物浓度为2-3mg/mL,PVA浓度为10-12%(w/v),AgNO3溶液浓度为2-3mg/mL, CS浓度为2-3%(w/v),Fe-PMs浓度为0.1-0.5mg/mL;
所述的石榴皮提取物按以下方法制备:将干燥石榴皮粉碎,过40-60目筛,称取石榴皮粉末0.1-0.2g,加入50-60mL 40%-50%乙醇,在50℃条件下超声25-30min,离心,取上清液,烘箱60-70℃烘1-2h后,得石榴皮提取物,根据需求用纯水配制为所需浓度。
2.如权利要求1所述的纳米酶可注射水凝胶在制备治疗痔疮的药物中的应用。
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Citations (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1066390A (zh) * | 1991-05-08 | 1992-11-25 | 安阿玥 | 治疗痔疮用注射液的制备方法 |
CN1718230A (zh) * | 2005-04-22 | 2006-01-11 | 张海峰 | 一种治疗痔疮的中药冻干粉针剂及其制备方法 |
CN101481714A (zh) * | 2008-01-11 | 2009-07-15 | 北京化工大学 | 一种以石榴皮为原料酶法制备鞣花酸的方法 |
CN108395440A (zh) * | 2018-04-28 | 2018-08-14 | 贵阳倍隆生物科技有限公司 | 一种利用没食子酸衍生物合成鞣花酸的方法 |
CN109082303A (zh) * | 2018-09-11 | 2018-12-25 | 福州大学 | 一种fcc汽油清洁化方法 |
CN115006586A (zh) * | 2022-01-24 | 2022-09-06 | 昆明理工大学 | 一种纳米酶原位水凝胶的制备方法及应用 |
CN115227864A (zh) * | 2022-07-23 | 2022-10-25 | 昆明理工大学 | 一种多功能纳米酶原位水凝胶的制备方法及应用 |
CN115386105A (zh) * | 2022-08-26 | 2022-11-25 | 昆明理工大学 | 多重酶活性纳米酶荧光水凝胶的制备方法及应用 |
CN115501335A (zh) * | 2022-08-30 | 2022-12-23 | 昆明理工大学 | 光声增强多重酶活性纳米酶水凝胶的制备方法和应用 |
-
2023
- 2023-06-02 CN CN202310650210.4A patent/CN116370406B/zh active Active
Patent Citations (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1066390A (zh) * | 1991-05-08 | 1992-11-25 | 安阿玥 | 治疗痔疮用注射液的制备方法 |
CN1718230A (zh) * | 2005-04-22 | 2006-01-11 | 张海峰 | 一种治疗痔疮的中药冻干粉针剂及其制备方法 |
CN101481714A (zh) * | 2008-01-11 | 2009-07-15 | 北京化工大学 | 一种以石榴皮为原料酶法制备鞣花酸的方法 |
CN108395440A (zh) * | 2018-04-28 | 2018-08-14 | 贵阳倍隆生物科技有限公司 | 一种利用没食子酸衍生物合成鞣花酸的方法 |
CN109082303A (zh) * | 2018-09-11 | 2018-12-25 | 福州大学 | 一种fcc汽油清洁化方法 |
CN115006586A (zh) * | 2022-01-24 | 2022-09-06 | 昆明理工大学 | 一种纳米酶原位水凝胶的制备方法及应用 |
CN115227864A (zh) * | 2022-07-23 | 2022-10-25 | 昆明理工大学 | 一种多功能纳米酶原位水凝胶的制备方法及应用 |
CN115386105A (zh) * | 2022-08-26 | 2022-11-25 | 昆明理工大学 | 多重酶活性纳米酶荧光水凝胶的制备方法及应用 |
CN115501335A (zh) * | 2022-08-30 | 2022-12-23 | 昆明理工大学 | 光声增强多重酶活性纳米酶水凝胶的制备方法和应用 |
Non-Patent Citations (1)
Title |
---|
金属基纳米酶的抗菌性能研究;朱军润;中国优秀硕士学位论文全文数据库工程科技Ⅰ辑;全文 * |
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