焦性没食子酸联合唑类药物对念珠菌的抑菌作用

目的了解光滑念珠菌临床菌株的药敏情况以及中药单体焦性没食子酸联合唑类药物对念珠菌的抑菌作用。方法收集临床分离的光滑念珠菌116株、白念珠菌49株、热带念珠菌42株、克柔念珠菌4株和近平滑念珠菌13株,采用ATB FUNGUS3药敏试条检测光滑念珠菌的药敏情况;同时采用棋盘肉汤稀释法检测焦性没食子酸联合唑类药物对念珠菌的抑菌情况。结果116株光滑念珠菌中,14.66%(17株)的菌株对氟康唑耐药,22.41%(26株)对伊曲康唑表现为非野生型和81.03%(94株)对伏立康唑表现为非野生型。焦性没食子酸对5种念珠菌的抑菌情况,46.55%光滑念珠菌的MIC值为64μg/mL;34.69%白念珠菌的MIC值为64μg/mL;59.52%热带念珠菌的MIC值为64μg/mL;25%克柔念珠菌的MIC值为128μg/mL;46.15%近平滑念珠菌的MIC值为128μg/mL。唑类药物与焦性没食子酸联合用药时,100%、99.14%、99.14%的光滑念珠菌分别对氟康唑、伊曲康唑和伏立康唑表现为协同作用或相加作用,差异无统计学意义(P>0.05);而白念珠菌、热带念珠菌、克柔念珠菌和近平滑念珠菌均表现为无关作用和拮抗作用。与单独用药相比,联合用药时81.03%、68.1%、77.59%的光滑念珠菌分别对氟康唑、伊曲康唑、伏立康唑的MIC值降低2~3个浓度梯度,且耐药组与非耐药组之间差异具有统计学意义(P<0.05)。结论光滑念珠菌对唑类药物具有耐药性,伏立康唑非野生型菌株所占比例最高。焦性没食子酸单独用药时,5组念珠菌中对光滑念珠菌的抑菌效果最好,且敏感组比耐药组的抑菌效果更加显著。焦性没食子酸联合唑类药物显著降低了光滑念珠菌唑类药物的MIC值,且耐药组比非耐药组的效果更加显著,为中西医结合治疗临床光滑念珠菌感染提供实验依据。     

五环三萜类柴胡皂苷单体对白念珠菌伊曲康唑耐药株活性研究

目的 从柴胡中提取具有抗真菌活性的五环三萜类单体Bp3,研究该单体对伊曲康唑耐药白念珠菌菌株的作用及其与伊曲康唑的相互作用.方法 参照CLSI标准,采用棋盘微量稀释法,测定单用Bp3、伊曲康唑及两者联合使用时对20株伊曲康唑耐药白念珠菌菌株的MIC,计算部分抑菌浓度指数(FIC),判定两药相互作用.结果 单独用药时Bp3及伊曲康唑对伊曲康唑耐药白念珠菌菌株MIC的几何均数值(GM值)分别为1.941 μg/mL和1.008 μg/mL.联合用药时Bp3和伊曲康唑的GM值分别降低为1.189 μg/mL和0.346 μg/mL.2种药物单用和联合使用时MIC值差异均有统计学意义(P＜0.05),联用时在20株耐药株均表现为协同或相加作用.结论 五环三萜类单体Bp3对白念珠菌伊曲康唑耐药株有一定的抑制作用,且与伊曲康唑有协同或相加作用.     

卡泊芬净、米卡芬净对念珠菌体外药物敏感性的动态研究

目的 动态研究卡泊芬净、米卡芬净体外对念珠菌的药物敏感性.方法 参照CLSI公布的M-27A方案微量液体稀释法分别测定卡泊芬净、米卡芬净、氟康唑对85株念珠菌的体外敏感性,并连续7d观测结果.结果 48 h卡泊芬净对白念珠菌、光滑念珠菌及其他念珠菌MIC50、MIC90中位数分别为0.030μg/mL、0.030 μg/mL,0.060μg/mL、0.125 μg/mL,0.125 μg/mL、0.500 μg/mL.48 h米卡芬净对白念珠菌、光滑念珠菌及其他念珠菌MIC50、MIC90中位数分别为0.030 μg/mL、0.030 μg/mL,0.060 μg/mL、0.060 μg/mL,0.250 μg/mL、0.500 μg/mL.48 h氟康唑对白念珠菌、光滑念珠菌及其他念珠菌MIC80、MIC100中位数分别为2μg/mL、128 μg/mL,64 μg/mL、128 μg/mL,2μg/mL、32μg／mL.85株念珠菌中未见对3种药物同时耐药的菌株.卡泊芬净组白念珠菌MIC50、MIC90 24 h后不再升高;光滑念珠菌MIC50 72 h后不再升高,MIC90 120 h后不再升高;其他念珠菌组MIC50 168 h、MIC90 96 h后不再升高.米卡芬净组白念珠菌、光滑念珠菌MIC50、MIC90 24 h后不再升高;其他念珠菌MIC50、MIC90在72 h后不再升高.结论卡泊芬净、米卡芬净对念珠菌属有较好的抗菌作用,其中对白念珠菌、光滑念珠菌作用更强,且MICs随着作用时间延长而升高并存在药物特异性和念珠菌种属特异性.     

基于蜡螟感染模型研究亚抑菌浓度氟康唑对光滑念珠菌毒力的影响

目的 基于蜡螟感染模型探究亚抑菌浓度(sub-MIC)氟康唑(fluconazole, FLU)对光滑念珠菌毒力的影响。方法收集青海省人民医院2021年4月至2022年10月临床分离非重复的光滑念珠菌作为实验菌株,采用ATB法测定FLU对光滑念珠菌的最低抑菌浓度(MIC)。采用蜡螟感染模型检测光滑念珠菌的毒力,并采用病理学和病原学方法评估建模是否成功。穿刺及不同sub-MIC FLU对健康蜡螟的影响：对照组不做任何处理,实验组分为三组,即PBS组(注射10μL PBS)、FLU亚组(分别注射10μL FLU药液,使感染蜡螟体内FLU浓度为0.5 mg/kg、1 mg/kg、2 mg/kg、4 mg/kg、8 mg/kg、16 mg/kg)和穿刺组(微量注射器穿刺但不注射)。不同sub-MIC的FLU对光滑念珠菌感染蜡螟的影响：采用2.50×10⁶酵母/只剂量感染蜡螟,2 h后,对照组再注射10μL PBS,实验组再注射不同浓度FLU工作液,观测对光滑念珠菌感染的蜡螟的治疗效果。结果 FLU对9株临床分离的光滑念珠MIC为8～32 mg/L,光滑念珠菌ATCC200...     

棘白菌素对氟康唑耐药的念珠菌体外药物敏感性的研究

目的评价3种棘白菌素类药物（卡泊芬净、米卡芬净、阿尼多芬净）体外对氟康唑耐药念珠菌的药物敏感性。方法采用微量液体稀释法和琼脂稀释法测定最小抑制浓度（MIC）。结果微量液体稀释法：59株耐药白念珠菌3种药物MIC50均为0.06μg/mL,米卡芬净、阿尼多芬净的MIC范围均为0.015～0.125μg/mL,卡泊芬净为0.015～0.25μg/mL;8株耐药光滑念珠菌MIC值均为0.063μg/mL。琼脂稀释法：59株耐药白念珠菌和8株耐药光滑念珠菌3种药物MIC值均为0.063μg/mL。结论3种棘白菌素类药物可能具有治疗氟康唑耐药的念珠菌感染的临床价值。     

289株VVC致病菌的菌种分布和对9种抗真菌药物敏感性

外阴阴道念珠菌病（vulvovaginal candidiasis,VVC）是女性的常见病。本研究收集了2018年1月-12月苏州地区VVC患者分离的289株念珠菌进行了病原学鉴定和包括棘白菌素类、新三唑类药物在内的9种抗真菌药物体外敏感性分析。本文采用核糖体RNA的D1/D2基因进行念珠菌菌种鉴定。参照M27-A3方法检测其对9种抗真菌药物（包括棘白菌素类及新三唑类药物）的体外敏感性。结果表明,289株VVC念珠菌菌株中,白念珠菌259株、光滑念珠菌14株、克柔念珠菌10株、热带念珠菌4株、近平滑念珠菌2株。259株VVC白念珠菌对棘白菌素类体外敏感性好,对米卡芬净敏感性高于另外两种棘白菌素类;对两性霉素B、5-氟胞嘧啶、氟康唑敏感性好;但对伊曲康唑、伏立康唑敏感性差;对泊沙康唑敏感性好。光滑念珠菌株和克柔念珠菌分离株对卡泊芬净敏感性差,但对米卡芬净、阿尼芬净敏感性好;光滑念珠菌株对两性霉素B、5-氟胞嘧啶体外敏感性好,对伊曲康唑敏感性差,对泊沙康唑敏感性好;伏立康唑对光滑念珠菌分离株MIC₅₀/₉₀为0.5/1μg/mL;克柔念珠菌对伊曲康唑、伏立康唑50%耐药;4株热带念珠菌对伊曲康唑50%耐药,对卡泊芬净、氟康唑、伏立康唑100%耐药,对其余5种抗真菌药物敏感。近平滑念珠菌对9种抗真菌药物均敏感。白念珠菌仍为苏州地区VVC的主要病原菌,其次是光滑念珠菌和克柔念珠菌,它们对临床常用药物伊曲康唑、伏立康唑、卡泊芬净敏感性差。研究结果提示对VVC病人常规进行分泌物培养、菌种鉴定,对苏州地区临床医生制定VVC治疗方案具有重要参考价值。尽管棘白菌素类、两性霉素B、5-氟胞嘧啶、新三唑类药物尚未应用到VVC的临床治疗中,但是这些药物对VVC病原体总体敏感性较好,未来有望成为氟康唑、咪唑类药物治疗失败患者的新选择。     

特康唑对常见念珠菌的体外敏感性试验

目的：研究特康唑对重度念珠菌性阴道炎常见念珠菌的体外敏感性。方法菌株分离自上海交通大学医学院附属仁济医院、上海市第六人民医院、上海市第一妇婴保健院妇科门诊外阴阴道念珠菌病（ VVC）患者的阴道分泌物。所有菌株均经念珠菌显色培养基（ CHROMagar Candida）、API 20C AUX鉴定,并采用CLSIM27?A3肉汤微量稀释法对213株临床分离念珠菌作了特康唑、克霉唑、咪康唑3种抗真菌药物敏感性测定。结果213株菌株经鉴定为白念珠菌192株（90．14％）,光滑念珠菌21株（9．86％）。特康唑对192株白念珠菌的MIC范围：0．024～12．5μg／mL,MIC50：0．39μg／mL,MIC90：1．56μg／mL,GM：0．248μg／mL;特康唑对19株光滑念珠菌的MIC范围：0．024～1．56μg／mL,MIC50：0．049μg／mL,MIC90：1．56μg／mL,GM：0．107μg／mL。结论目前阴道念珠菌病的病原菌仍以白念珠菌为主,并提示特康唑与克霉唑、咪康唑3种药物对213株念珠菌均具有良好的敏感性,特康唑对白念珠菌及光滑念珠菌有较强的抑制作用并未发现有耐药现象。     

新疆地区白念珠菌基因型分析及其体外药物敏感性研究

目的研究新疆地区汉族和维吾尔族患者来源的50株白念珠菌的基因型及其对两性霉素B、5-氟胞嘧啶、米卡芬净、伊曲康唑、氟康唑和咪康唑的体外敏感性。方法采用PCR法扩增白念珠菌rDNA 25S的Ⅰ类内含子包含区,根据扩增产物的大小判断基因型(A型为450 bp,B型为840 bp,C型为450 bp和840 bp)。采用CLSI M27-A液基微量稀释法测定50株白念珠菌对上述6种抗真菌药的体外敏感性。结果 50株菌分为3种基因型:A型30株,B型和C型各10株。所有菌株对两性霉素B、5-氟胞嘧啶、米卡芬净和咪康唑的MIC值较低,MIC范围依次为0.25～0.5μg/mL,0.125～0.5μg/mL,≤0.03μg/mL,0.25～8μg/mL;对伊曲康唑和氟康唑的MIC值较高,MIC范围分别为0.25～8μg/mL,0.5～64μg/mL。B型和C型对5-氟胞嘧啶的MIC值均为0.125μg/mL,对伊曲康唑和氟康唑的耐药率分别为84%、70%。不同族别来源的菌株基因型比较无显著差异(P>0.05),不同基因型菌株的抗真菌药物敏感性比较也无显著差异(P>0.05)。结论新疆地区白念珠菌分A,B,C三种基因型。汉...     

复发性外阴阴道念珠菌病的致病菌种鉴定及药敏分析

目的:探究复发性外阴阴道念珠菌病致病菌的菌种分布及耐药情况,指导临床治疗。方法:收集210例念珠菌性外阴阴道炎患者阴道分泌物标本,其中复发性外阴阴道念珠菌病(RVVC)组76例,外阴阴道念珠菌病(VVC)组134例。运用科玛嘉显色培养基法进行菌种鉴定和药物敏感试验。结果:在210例病例中,RVVC组中白色念珠菌54株(71.05%),光滑念珠菌15株(19.74%),热带念珠菌4株(5.26%),克柔念珠菌2株(2.63%),近平滑念珠菌1株(1.32%)。非白色念珠菌中以光滑念珠菌为主,在RVVC组中的比例明显高于VVC组,有显著性差异(P<0.05)。药敏试验显示,RVVC的念珠菌株中仅8株对7种药物全部敏感,23株敏感药物<5种,其中1株仅对制霉菌素敏感;对药物的敏感率为制霉菌素(97.37%)>克霉唑(60.52%)>酮康唑(51.32%)>伊曲康唑(36.84%)>咪康唑(35.53%)>氟康唑(23.68%)>特比奈芬(10.53%)。结论:白色念珠菌仍是RVVC的主要致病菌,非白色念珠菌比例在RVVC组中显著高于VVC组,其中以光滑念珠菌为主。念珠菌对制霉菌素敏感率最高,对唑类药物耐药性有增加趋势。     

米卡芬净对分离自中国的念珠菌和曲霉临床株体外抑菌活性的研究

目的了解新型抗真菌药物米卡芬净(micafungin,MFG)对分离自中国的念珠菌和曲霉临床株的体外抑菌活性。方法参照CLSI(Clinical and Laboratory Standards Institute,以前为NCCLS)制定的M27-A2和M38-A方案测定86株念珠菌和35株曲霉的最低抑菌浓度(MIC)或最低有效浓度(MEC)。结果MFG对大多数念珠菌属和曲霉属均有较好的抑菌作用。对念珠菌属的MIC90从高到低依次为:氟康唑(FLC)敏感的白念珠菌、热带念珠菌、光滑念珠菌为0.125μg/ml,FLC耐药和剂量依赖敏感株为0.25μg/ml,克柔念珠菌为0.5μg/ml,近平滑念珠菌8μg/ml,季也蒙念珠菌>16μg/ml。MFG对烟曲霉的MEC90为≤0.03μg/ml,对非烟曲霉的曲霉属MEC90为0.06μg/ml。MFG与唑类药物、两性霉素B(AMB)不存在交叉耐药,对FLC耐药的念珠菌、伊曲康唑耐药的曲霉、AMB不敏感的曲霉均有好的抑菌活性。结论MFG对多数念珠菌属和曲霉属(包括对唑类耐药和AMB不敏感的菌株)有较好的体外抑菌作用。     

耐氟康唑念珠菌和耐伊曲康唑烟曲霉对泊沙康唑的敏感性测定

目的：测定耐氟康唑念珠菌和耐伊曲康唑烟曲霉临床分离株对泊沙康唑的敏感性。方法参照美国临床实验室标准化研究所制定的 M27-A3和 M38-A2方案,测定从临床获得的11株耐氟康唑的念珠菌和3株耐伊曲康唑烟曲霉对泊沙康唑的 MIC 值。结果对于氟康唑耐药的念珠菌,泊沙康唑的 MIC 范围是0.125-1μg/ mL。对于伊曲康唑耐药烟曲霉,泊沙康唑的 MIC 范围是0.06-0.5μg/ mL。结论11株耐氟康唑的念珠菌和3株耐伊曲康唑烟曲霉均对泊沙康唑有效。     

Synergic effect of metronidazole and pyrantel pamoate on Giardia lamblia

Giardia lamblia is a pathogenic protozoan presenting as the main characteristic, the trophozoite capacity to adhere in host intestinal epithelium, infecting mammals, including humans. The clinical treatment of this disease is based on metronidazole (Mz) that acts as an alternative electron acceptor, and its reduction promotes DNA impairment. In veterinary treatment, one of the best options is pyrantel pamoate (Pm), which the mode of action has not elucidated yet. Different strategies for Giardia treatment have been explored to avoid side effects to the host. In this context, the efficiency of treatment combining drugs raise as an interesting alternative for protozoan diseases. Here, we evaluated in vitro synergic effect of Mz and Pm on trophozoites and on its adherence to IEC-6 cells. The treatment with Mz or Pm was effective on trophozoites, with IC(50)/24h values of 5.3±0.9 μM and 13.8±1.4 μM, respectively. The treatment of trophozoites with different combinations of Mz and Pm were also evaluated, as showed by fractional inhibitory concentration index (FICI) under 0.5 in all conditions tested, corresponding to the synergic effect. This synergic activity was also observed when the combinations of 5.3 μM Mz+0.4 μM Pm and 13.8 μM Pm+0.1 μM Mz induced a remarkable reduction in % adhesion (85-90% and 52-59%, respectively) and in number of adhered parasites per 100 cells. The low cytotoxicity to the host cells of the combinations, associated to the strong synergic potential of the combination, encourage us to further investigate its effect in in vivo models.     

Synergistic effects between silibinin and antibiotics on methicillin-resistant Staphylococcus aureus isolated from clinical specimens

Methicillin-resistant Staphylococcus aureus (MRSA) is a dangerous microorganism, and creates serious medical problems. It causes many types of infections in humans and often acquires multi-drug resistance. In this study, silibinin was evaluated against 20 clinical isolates of MRSA, either alone or in combination with ampicillin or oxacillin, using a checkerboard assay. The silibinin exhibited good activity against isolates of MRSA, and MRSA ATCC33952 and MSSA ATCC25923, with minimum inhibitory concentrations/minimum bactericidal concentrations (MICs/MBCs) ranging between 2-8/4-16 μg/mL, for ampicillin 2-1024/2-2048 μg/mL, and for oxacillin 0.25-32/0.5-64 μg/mL. The range of MIC(50) and MIC(90) were 0.5-4 μg/mL and 2-8 μg/mL, respectively. The MICs/MBCs for the combination of silibinin plus oxacillin or ampicillin were reduced by ≥4-fold against the MRSA isolates tested, demonstrating a synergistic effect, as defined by a fractional inhibitory concentration index (FICI) of ≤0.5. Furthermore, a time-kill study evaluating the growth of the tested bacteria showed that growth was completely attenuated after 2-5 h of treatment with the 1/2 MIC of silibinin, regardless of whether it was administered alone or with oxacillin (1/2 MIC) or ampicillin (1/2 MIC). In conclusion, silibinin exerted synergistic effects when administered with oxacillin or ampicillin and the antibacterial activity and resistant regulation of silibinin against clinical isolates of MRSA might be useful in controlling MRSA infections.     

覆盆子提取物联合唑类药物抗真菌活性研究

目的 探讨中药覆盆子提取物联合唑类药物的体外抗真菌作用.方法 采用CLSI公布的M27-A方案微量液基稀释法和棋盘式微量稀释法,测定覆盆子提取物单用及联合唑类药物对不同念珠菌的MIC值和FICI指数.结果 覆盆子不同溶液提取物与氟康唑均表现出协同关系,以覆盆子醇提物为例,单用对念珠菌的MIC80测定值范围主要集中在0.16～1.25 mg/mL,与氟康唑合用后表现出协同关系(FICI≤0.5),且MIC80测定值范围降至0.01 ～0.04 mg/mL;合用后的氟康唑抗真菌活性也明显增强.另外,覆盆子醇提物与不同唑类药物合用后均有协同关系,其MIC80测定值由单用时大于10 mg/mL降至0.04 mg/mL.结论 覆盆子醇提物和唑类药物单用时对耐药念珠菌的抑菌作用较弱,但二者合用后表现出明显的协同关系,对耐药念珠菌的抑菌作用明显增强.     

Fungicidal activity of the human peptide hepcidin 20 alone or in combination with other antifungals against Candida glabrata isolates

Candida glabrata infections are often difficult to eradicate due to the intrinsically low susceptibility to azoles of this species. In addition, C. glabrata has also been shown to be insensitive to several cationic peptides, which have been shown to be promising novel therapeutic candidates for the treatment of fungal infection. In this study, the in vitro fungicidal activity of the human cationic peptide hepcidin 20 (Hep-20) was evaluated against clinical isolates of C. glabrata with different levels of fluconazole susceptibility. Interestingly, all isolates were susceptible to Hep-20 (100–200 μg/ml) at pH 7.4, whereas the fungicidal effect of the peptide was higher (50 μg/ml) at acidic pH values. In addition, an increased antifungal activity was observed for Hep-20 with amphotericin B and a synergistic effect was demonstrated for the Hep-20/fluconazole and Hep-20/caspofungin combinations.     

Antifungal effects of Ellagitannin isolated from leaves of Ocotea odorifera (Lauraceae)

Ocotea odorifera is a medicinal plant that is popularly known in Brazil as "canela-sassafrás" and is used to treat dermatosis. This study investigated the antifungal properties of O. odorifera. The methanol extract of O. odorifera was submitted to successive chromatographic separation and yielded Tellimagrandin II (TEL). Candida parapsilosis strain ATCC 22019 was used to determine the minimum inhibitory (MIC) and fungicidal concentrations, and to study the synergistic action with nystatin (NYS), amphotericin (AMP), and fluconazole (FLU). After treatment, the morphology of the yeast was analysed by scanning electron microscopy. Cytotoxicity was assessed in Vero cells, and genotoxicity by the micronucleus test. The TEL structure was proposed based on NMR and comparison with literature data and ESI-MSMS analysis. The compound showed potent inhibitory activity against C. parapsilosis, with a MIC of 1.6 μM. TEL acted synergistically with NYS, AMP, and FLU, and caused morphological alterations in the yeast cells. The methanolic extract showed low cytotoxicity in vitro and in vivo, and was not mutagenic in mice (P < 0.05). The use of O. odorifera in traditional medicine seems to have a valid basis, in view of the antifungal activity of TEL demonstrated in this study, and may contribute to potential drug development.     

Design,synthesis and biological evaluation of novel semicarbazone-selenochroman-4-ones hybrids as potent antifungal agents

A series of novel 2,3-dihydro-4H-1-benzoselenin-4-one (thio)semicarbazone derivatives were designed and synthesized by using molecular hybridization approach. All the target compounds were characterized by HRMS and NMR and evaluated in vitro antifungal activity against five pathogenic strains. In comparison with precursor selenochroman-4-ones, the hybrid molecules in this study showed significant improvement in antifungal activities. Notably, compound B8 showed significant antifungal activity against other strains excluding Aspergillus fumigatus (0.25 μg/mL on Candida albicans, 2 μg/mL on Cryptococcus neoformans, 8 μg/mL on Candida zeylanoides and 2 μg/mL on fluconazole-sensitive strains of Candida albicans). Moreover, compounds B8, B9 and C2 also displayed most potent activities against four fluconazole-resistance strains. Especially the MIC values of the hybrid molecule B8 against fluconazole-resistant strains were in the range of 0.5–2 μg/mL. Therefore, the molecular hybridization approach in this study provided new ideas for the development of antifungal drug.