结核分枝杆菌喹诺酮类药物耐药与基因突变关系的研究

目的:探讨喹诺酮类药物的耐药性与结核分枝杆菌gyrA基因突变的关系。方法:采用比例法检测结核分枝杆菌中氧氟沙星耐药的菌株,并对氧氟沙星耐药菌株的gyrA基因的耐药决定区域进行序列测定。结果:175株结核分枝杆菌中,耐氧氟沙星菌株为30.9%(54/175),其中81.5%(44/54)的菌株gyrA基因的喹诺酮耐药决定区发生突变。结论:在结核病人中存在着较严重的耐喹诺酮药物情况,因此对结核病人选择喹诺酮药物治疗时,最好先进行喹诺酮药物的耐药检测,gyrA基因突变的检测可预测菌株对喹诺酮类药物的耐药表型。     

从胡椒属植物diospyrifolium中分离得到1种新的苯甲酸衍生物及其抗结核分枝杆菌活性检测

作者采用超临界流体法(SFE-CO2)和色谱法，从胡椒属植物 diospyrifolium 叶中分离鉴定出1种新的苯甲酸衍生物4-甲氧基-3-[(E)-3-甲基-1,3-丁二烯-1-yl]-5-(3-甲基-2-丁烯-1-yl)-苯甲酸(1)。在巨噬细胞J774G8中，对SFE-CO2提取物和化合物1的抗结核分枝杆菌活性和细胞毒性进行了检测。化合物1和SFE-CO2提取物显示了中度抗结核分枝杆菌 H37Rv 的活性，其最小抑菌浓度(MIC)值为125μg/ml。结核分枝杆菌临床分离菌株的MIC值范围125～250μg/ml。细胞毒性检测结果显示的选择性指数范围为0.6～1.0。对此化合物应进一步研究其结构与活性的关系，以及与抗结核药物的协同作用，以便更好的评估此化合物的抗结核分枝杆菌活性。     

广东顺德区结核分枝杆菌临床耐药性分析

目的:了解本地区结核分枝杆菌临床耐药性的特点以及对7种抗结核药物的药敏检测情况,为临床制定合理的治疗方案提供更多的参考依据。方法:对收集的广东顺德区555例结核患者的结核分枝杆菌用比例法对所用的7种药:异烟肼(INH,H)、乙胺丁醇(EMB,E)、链霉素(SM,S)、利福平(RFP,R)、阿米卡星(AKW,A)、左氧氟沙星(LOF,L)、对氨基水杨酸(PAS,P)进行敏感性试验,分析它们的耐药性。结果:总耐药率为33%,耐药频度由高到低依次为SM19.8%、INH12.8%、LOF8.6%、RFP4.5%、EMB3.6%、PAS3.1%、AKW1.4%。结论:本地结核菌的耐药性相比其他地区略偏高,应进一步加强对结核杆菌的耐药性监测以便为临床制定更合理的治疗方案提供参考依据。     

银环蛇蛇毒中具有镇痛活性的多肽成分的生化分离和活性鉴定

目的 研究银环蛇（Bungarus multicinctus）毒的镇痛活性，并对其中的活性单体进行分离、纯化与鉴定。方法 采用热痛、醋酸扭体动物模型检测银环蛇（Bungarus multicinctus）蛇毒毒的镇痛作用，通过分子筛、高效液相（HPLC）等纯化手段从银环蛇毒中分离出镇痛单体，然后通过肽谱鉴定，氨基酸测序及序列比对鉴定其结构特征。结果 本研究发现银环蛇粗毒具有显著的镇热痛活性，与生理盐水组相比，热痛阈提高近2℃。与吗啡相比，其镇痛作用持续时间更长，在给药7 h后仍有显著镇痛效果。通过分子筛及高效液相分离，我们发现其镇痛活性成分主要为小分子多肽，其中活性最强的成分是α-bungarotoxin。α-bungarotoxin有与银环蛇粗毒类似的热镇痛活性，并且能够显著抑制醋酸导致的急性炎性痛。序列鉴定及结构分析表明α-bungarotoxin是一种含有74个氨基酸，5对二硫键的三指肽，能够与乙酰胆碱受体结合。 结论 银环蛇（Bungarus multicinctus）蛇毒具有显著的镇痛作用。其中作用于乙酰胆碱受体的三指肽α-bungarotoxin是其最主要的镇痛活性成分。     

应用体外抑菌法筛选中华大蟾蜍抗结核有效成分的研究

目的验证中华大蟾蜍具有抗结核活性，筛选抗结核活性成分。方法对中华大蟾蜍水提物进行葡聚糖凝胶分子筛分离和反相高效液相层析法（RPLC）分离，并采用微孔板Alamarblue法将分离所得12组分进行体外抑菌实验研究。结果第1、6、10组分表现出对供试结核分枝杆菌的明显抑制作用，其最低抑菌浓度（minimal inhibitory concentration，MIC）分别为339．98mg／ml、20．00mg／ml、2．95mg／ml。结论中华大蟾蜍含有抑制结核分枝杆菌的活性成分，该有效成分分子量接近14KD，并表现出弱极性的特征。     

牛贝消核提取物对耐多药结核分枝杆菌和非结核分枝杆菌的体外抑菌作用的研究

目的研究不同工艺制备的中药复方牛贝消核提取物的体外抑菌活性,为制备抗结核中成药制剂提供实验基础。方法制备分别含50,100,200 g·L-1牛贝消核水提取物、醇提取物的改良罗氏培养基,并分别制备含1,10μg/ml异烟肼(Isonicotinic acid hydrazide,INH)和50,250μg·ml-1利福平(Rifampicin,RFP)的改良罗氏培养基为对照,将3株耐多药结核分枝杆菌分离株、牛结核分枝杆菌和4株非结核分枝杆菌分别接种于含药的改良罗氏培养基中,置37℃培养4周,测定其最低抑菌浓度(MICs)。结果牛贝消核水提取物对这8株供试菌的MICs均为200 g·L-1;醇提取物对牛结核分枝杆菌和母牛分枝杆菌的MICs均为100 g·L-1,对3株耐多药结核分枝杆菌分离株、胞内、瘰疬、偶发分枝杆菌的MICs均为200 g·L-1。结论牛贝消核水提取物和醇提取物对敏感或耐药的结核分枝杆菌和非结核分枝杆菌均有不同程度的抑制作用,醇提取物的抑菌活性比水提取物略强。     

结核分枝杆菌抗体在肺结核患者中的临床研究

目的:应用痰涂片查抗酸杆菌、胶体金法检测结核分枝杆菌抗体、X线胸片三种方法,对诊断肺结核进行对比评估。方法:对我院确诊的108例肺结核患者的血清进行结核分枝杆菌抗体检测,同时做痰涂片查抗酸杆菌及X胸片检查。结果:采用血清进行结核分枝杆菌抗体检测有78例阳性,X胸片检查有84例阳性,痰涂片查抗酸杆菌有46例阳性。以胶体金法测结核分枝杆菌抗体与X线胸片两法具有较好的敏感性,差异无统计学意义(P>0.05)。在108例认定肺结核的标本进行检测结果分析,痰涂片阳性率为42.6%与结核分枝杆菌抗体的阳性率为72.2%和X线胸片阳性率为77.8%差异均具有统计学意义(P<0.05)。结论:血清结核分枝杆菌抗体检测在肺结核诊断中具有方便、快速、敏感、特异的特点,结合X胸片检查具有更高的临床应用价值。     

蝮蛇蛇毒中小分子肽的纯化及其抑制血小板聚集的作用

本文首次报道从中国蝮蛇蛇毒中分离出一种分子量为1234.616Da的具有抑制血小板聚集作用的小肽SV-PP-1.SV-PP-1能明显抑制ADP诱导的兔血小板聚集,并有剂量依赖性.这是目前从蛇毒中发现的分子量最小的能抑制血小板聚集的蛇毒小肽.     

耐异烟肼结核杆菌感染小鼠动物模型的建立与评价

目的:建立耐异烟肼结核分枝杆菌尾静脉注射感染小鼠模型。方法:将30只昆明种SPF级小鼠随机分为空白组、耐异烟肼组、标准菌株组,每组10只,通过尾静脉注射,分别接种同剂量生理盐水、耐异烟肼菌株混悬液、标准菌株混悬液,观察一般情况,1周后处死小鼠,进行细菌培养,观察菌落生长情况。结果:空白组和标准菌株组造模后体质量增加,耐异烟肼菌株组造模后体质量无显著增加。造模后的耐异烟肼菌株组、标准菌株组小鼠肺组织均出现小结节。细菌培养可见空白对照组未培养出结核菌,耐异烟肼菌株组、标准菌株组中小鼠肺组织均培养出结核分枝杆菌。结论:成功建立了耐异烟肼结核分枝杆菌静脉注射感染小鼠模型,为进一步研究耐异烟肼结核杆菌奠定基础。     

172株结核分枝杆菌耐药性分析

目的,了解我县肺结核病人对常用抗结核药物的敏感情况。方法,利用高低两个药物浓度分别检测172株结核分枝杆菌对异烟肼（INH）、链霉素（SM）、乙胺丁醇（EMB）、利福平（RFP）的耐药性。结果,172株结核分枝杆菌全敏感81株占47．1％,耐药91株占52．9％。结论单一耐药高于同时耐两种药,同时耐两种药高于耐三种药,低浓度耐药高于高浓度耐药。     

In vitro antimicrobial and antioxidant activities of anthrone and chromone from the latex of Aloe harlana Reynolds

In the search for new antimicrobial and antioxidant compounds from plants, the latex of the medicinal plant Aloe harlana Reynolds from Ethiopia was subjected to bioassay‐guided fractionation, which led to the isolation of two known compounds, anthrone (aloin) and chromone (7‐O‐methylaloeresin A). The latex and its two constituents were assessed for their possible antimicrobial activities against 23 bacterial and four fungal strains using the disc diffusion method and their antioxidant activity by two complementary test systems, namely 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH) and 2‐deoxyribose degradation assay methods. The isolated compounds showed promising results against various pathogenic bacterial and fungal strains in comparison with standard drugs. Moreover, 7‐O‐methylaloeresin A exhibited good activity against multiple drug resistant Staphylococcus aureus (NCTC 11994) and Salmonella typhimurium (ATCC 1255) with MIC values of 0.72 and 0.18 mm , respectively. Among the fungal strains tested, Candida albicans (ATCC 10231) was the most susceptible organism to the latex and the two isolated compounds. The latex and isolated compounds also showed significant activities on both antioxidant assays with the highest activity being observed for 7‐O‐methylaloeresin A, which gave IC₅₀ values of 0.026 mm and 0.021 mm for DPPH and 2‐deoxyribose degradation assay, respectively. These findings support the traditional uses of the plant for the treatment of various infectious and inflammatory diseases. Copyright © 2011 John Wiley & Sons, Ltd.     

药用植物三尖杉内生真菌的分离鉴定及抗菌活性

目的：研究药用植物三尖杉Cephalotaxus fortunei内生真菌的分离鉴定及抗菌活性。方法：用组织培养方法从野生三尖杉的根、茎、叶中分离内生真菌,采用滤纸片扩散法对分离得到的内生真菌的代谢产物做抑菌活性研究。结果：分别从三尖杉根、茎、叶中共分离得到30株内生真菌;抑菌活性研究显示,有20株内生真菌的代谢产物至少对1种靶标菌有抑制活性,占总分离菌株的66.7%,其中菌株CFEF013对所有靶标菌均有抑制作用;经形态学和分子生物学鉴定,对7株具有较强抑菌活性的菌株分属镰孢菌属Fusarium 4株,壳囊孢属Cytospora 1株,拟盘多毛孢属Pestalotiopsis 1株,毛霉菌属Mucor 1株。结论：药用植物三尖杉内生真菌种类具有一定的多样性,内生真菌代谢产物抑菌活性良好。     

Antiplasmodial and cytotoxic activity of phloroglucinol derivatives from Hypericum erectum thunb

The chloroform extracts of whole plants of Hypericum erectum were investigated for antiplasmodial activity against chloroquine‐sensitive strains of Plasmodium falciparum. Five phloroglucinol derivatives, otogirin (1), otogirone (2), erectquione A (3), erectquione B (4), and erectquione C (5) were isolated from the whole plants of H. erectum. Also, five compounds were evaluated for in vitro antiplasmodial activities as well as their cytotoxic potential on SK‐OV‐3 cancer cell line cells. Compounds 2, 4 showed notable growth inhibitory activity against chloroquine‐sensitive strains of Plasmodium falciparum with IC₅₀ values from 5.6 and 7.2 μM. This compound showed no significant cytotoxicity (IC₅₀ > 150 μM) evaluated using SK‐OV‐3 cancer cell line cells. Copyright © 2010 John Wiley & Sons, Ltd.     

Antiprotozoal activity of drimane and coloratane sesquiterpenes towards Trypanosoma brucei rhodesiense and Plasmodium falciparum in vitro

The extracts and 12 sesquiterpenes obtained from the East African medicinal plant Warburgia ugandensis Sprague (Canellaceae) were assessed for their antiplasmodial activity against the chloroquine‐sensitive (3D7) and chloroquine‐resistant (K1) strains of Plasmodium falciparum and antitrypanosomal activity against Trypanosoma brucei rhodesiense. The dichloromethane extract displayed strong antiplasmodial and antitrypanosomal activities with IC₅₀ values of 8.10 and 1.10 µg/mL against K1 strain of the malaria parasite and STlB900 strain of T. b. rhodesiense, respectively. Among the compounds evaluated for inhibition of trypomastigotes, both drimane and coloratane sesquiterpenes possessing aldehyde groups at positions 8 and 9 were found to show most antitrypanosomal activity with IC₅₀ values in the range 0.56–6.4 µM. The antiplasmodial assays also revealed that the six coloratane and six drimane sesquiterpenes isolated from this extract exhibited significant antitrypanosomal activity with IC₅₀ values ranged from 0.45 to ?114 µM. Among the compounds tested against the malarial parasite P. falciparum 11?‐hydroxymuzigadiolide (3) was most active with an IC₅₀ value of 6.40 µM. Copyright © 2010 John Wiley & Sons, Ltd.     

益母草的化学成分及其抗人白血病K562细胞活性研究

目的:研究益母草的化学成分,评价益母草化学成分体外抗白血病K562细胞活性.方法:运用硅胶柱色谱、葡聚糖凝胶sephadex LH-20、聚酰胺柱色谱和大孔树脂等色谱方法对益母草化学成分进行分离纯化,利用核磁共振技术鉴定了它们的化学结构.通过测试体外最低抑制浓度评价有关物质的抗肿瘤活性.结果:分离、纯化并鉴定了8个化合物,分别为槲皮素-3-O-洋槐双糖苷(quercetin 3-O-robinobioside,1),芦丁(rutin,2),异槲皮苷(isoquercitrin,3),金丝桃苷(hyperoside,4),槲皮素(quercetin,5),芹菜素(apigenin,6),芫花素(genkwanin,7)和苯甲酸(benzoic.acid,8).结论:化合物1,3,4,8为首次从该属植物中分离得到,化合物2,5-7首次从该种植物中分离得到.化合物1-6,8显示出不同程度的抑制人白血病K562细胞活性.     

广藿香内生真菌类群分析及其抗菌活性研究

从广藿香的根、茎、叶中共分离得到72株内生真菌,根据ITS序列分析鉴定为25属40个种,其中拟茎点霉属Phomopsis、刺盘孢属Colletotrichum、镰刀菌属Fusarium为优势种群;广藿香内生真菌的分布存在明显的组织特异性,以茎中内生真菌的分布最多,占分离菌株总数的78%;活性测试结果显示共有15个属34株内生真菌至少对1种供试菌具有抗菌活性。结果表明广藿香内生真菌多样性丰富,部分菌株具有明显的抑菌活性,值得进一步深入研究。     

In vivo and in vitro antimalarial activity of 4-nerolidylcatechol

4‐Nerolidylcatechol (4‐NC) isolated from Piper peltatum L. (Piperaceae) was evaluated for in vitro antiplasmodial activity against Plasmodium falciparum (cultures of both standard CQR (K1) and CQS (3D7) strains and two Amazonian field isolates) and for in vivo antimalarial activity using the Plasmodium berghei‐murine model. 4‐NC exhibits significant in vitro and moderate in vivo antiplasmodial activity. 4‐NC administered orally and subcutaneously at doses of 200, 400 and 600 mg/kg/day suppressed the growth of P. berghei by up to 63% after four daily treatments (days 1–4). Also, 4‐NC exhibited important in vitro antiplasmodial activity against both standard and field P. falciparum strains in which 50% inhibition of parasite growth (IC₅₀) was produced at concentrations of 0.05–2.11 μg/mL and depended upon the parasite strain. Interestingly, healthy (non‐infected) mice that received 4‐NC orally presented (denatured) blood plasma which exhibited significant in vitro activity against P. falciparum. This is evidence that mouse metabolism allows 4‐NC or active metabolites to enter the blood. Further chemical and pharmacological studies are necessary to confirm the potential of 4‐NC as a new antimalarial prototype. Copyright © 2011 John Wiley & Sons, Ltd.     

温莪术内生真菌的分离及其抗菌活性的初步研究

 目的研究温莪术内生真菌的种群组成及其代谢产物抗菌活性。方法采用内生菌常规分离法对健康温莪术植株体内的内生真菌进行了分离鉴定。以大肠杆菌、金黄色葡萄球菌、链球菌、沙门氏菌、巴氏杆菌为测试菌种,对温莪术根、茎、叶中分离的56株内生真菌进行抗菌活性筛选。结果经初步鉴定,这些内生真菌为5目、6科、24属;抑菌实验表明,56株内生真菌中有39株对5种测试菌均有不同程度的抑制作用。结论温莪术植物含有丰富多样的内生真菌,且多数内生真菌都表现为较高的抑菌活性。     

Antiplasmodial and cytotoxic activity of khellactone derivatives from Angelica purpuraefolia chung

The methanolic extract of the rhizomes parts of Agelica purpuraefolia was investigated for its activity against chloroquine‐sensitive strains of Plasmodium falciparum using the parasite lactate dehydrogenase assay method. Two natural khellactone, (+)‐4′‐Decanoyl‐cis‐khellactone (1) and (+)‐3′‐Decanoyl‐cis‐khellactone (2) were isolated from the rhizomes parts of A. purpuraefolia. Two compounds were evaluated for in vitro antiplasmodial activities as well as their cytotoxic potential on SK‐OV‐3 cancer cell line cells. Compounds 1, 2 showed notable growth inhibitory activity against chloroquine‐sensitive strains of Plasmodium falciparum with IC₅₀ values from 1.5 and 2.4 μM. This compound showed no significant cytotoxicity (IC₅₀ > 100 μM) evaluated using SK‐OV‐3 cancer cell line cells. This is the first report on the antiplasmodial activity of the compounds from A. purpuraefolia. Copyright © 2009 John Wiley & Sons, Ltd.     

桃儿七内生真菌分离及其抑菌活性初步研究

目的 探明桃儿七内生真菌资源多样性及其抑菌活性特征.方法 通过组织块法对内生真菌进行分离,并选择5种植物病原真菌和4种细菌作为指示菌,采用对峙法和改进的菌块法测定抑菌活性.结果 从桃儿七根、茎、叶中分离出49株内生真菌,根部最多,22株,其次为茎部,18株,叶部最少,9株;经形态学分类鉴定归于2个目,3个科,9个属,梭孢霉属为优势菌属;有22株菌对1种或多种供试植物病原真菌有不同程度的抑制作用,占分离菌株数的44.8％,有8株菌对2种或多种供试植物病原真菌有明显的抑制作用,其占所分离菌株总数的16.3％,有3株菌对4种或多种供试植物病原真菌有明显抑制作用,2株内生真菌分别对2种供试指示细菌具有明显的抑制作用.结论 桃儿七内生真菌具有多样性和明显的抗外源真菌活性,抑菌活性菌株主要分布在曲霉属、单孢枝霉属、梭孢霉属,其内生真菌的研究和开发具有重要的生态和经济意义.