水飞蓟素脂质体对小鼠CCl4急性肝损伤保护作用的实验研究

目的：研究水飞蓟素脂质体（L-SIL）对四氯化碳（CCl4）所致小鼠急性肝损伤的保护作用.方法：昆明种小鼠80只,随机分为空白组、CCl4急性肝损伤模型组及L-SIL低、中、高剂量组和益肝灵（T-SIL）低、中、高剂量组共8组,测量并比较各组小鼠血清丙氨酸氨基转移酶（ALT）、天门冬氨酸氨基转移酶（AST）水平及肝组织匀浆中超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GSH-PX）水平和丙二醛（MDA）含量,并观察各组小鼠肝组织病理学改变.结果：与CCl4急性肝损伤模型组相比,L-SIL各剂量组血清ALT、AST水平明显降低（P〈0.05～0.01）,L-SIL各剂量肝组织匀浆GSH-PX水平及L-SIL高剂量组SOD水平明显升高（P〈0.05～0.01）,并具有剂量依赖性,L-SIL高剂量组肝组织匀浆MDA含量明显降低（P〈0.01）.L-SIL对ALT、SOD、GSH-PX水平的作用优于相应剂量T-SIL组（P〈0.05～0.01）.组织学观察L-SIL中、大剂量组的肝损伤明显轻于模型组和相应剂量T-SIL组.结论：L-SIL对小鼠CCl4肝损伤有明显的保护作用,且其保护作用明显优于T-SIL.     

水飞蓟素脂质体对BCG+LPS引起的小鼠免疫性肝损伤的保护作用

目的研究水飞蓟素脂质体(L-SIL)对卡介苗(BCG) 脂多糖(LPS)引起的小鼠免疫性肝损伤的保护作用.方法昆明种小鼠80只随机分为空白组、模型组及L-SIL低、中、高剂量组和益肝灵(T-SIL)低、中、高剂量组共8组,测量并比较各组小鼠血清丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)水平和肝组织匀浆中超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-PX)水平以及丙二醛(MDA)、一氧化氮(NO)含量,观察肝脏、脾脏系数的变化,并观察各组小鼠肝组织病理学改变.结果与模型组相比,L-SIL各剂量组血清ALT、AST水平明显降低(P＜0.05～0.01),肝组织匀浆GSH-PX、SOD水平明显升高(P＜0.05～0.01),MDA和NO含量明显降低(P＜0.01),肝脏、脾脏系数明显降低(P＜0.05～0.01).L-SIL对ALT、AST、GSH-PX水平和NO含量的作用强于同剂量T-SIL组(P＜0.05～0.01).组织学观察L-SIL低、中剂量组的肝损伤明显轻于模型组和相应剂量T-SIL组.结论L-SIL对小鼠BCG LPS引起的免疫性肝损伤有明显的保护作用,且其保护作用明显优于T-SIL.     

胃癌特异性免疫脂质体导向治疗的实验研究

将胃癌单克隆抗体ＭＧｂ２与脂质体连接，分别包人１１种抗肿瘤药物和硼１０化合物?制成９种胃癌特异性免疫脂质体和２种硼化胃癌免疫脂质体，采用不同的人胃癌裸鼠模型和不同的给药途径，综合评价其体内外疗效和生物学分布。９种免疫脂质体较好地保持了抗体的免疫活性，对胃癌细胞具有高度选择性伤作用。疗效均显著优于非特异脂质体和游离药物。硼化免疫脂质体结合热中子照射后显示出导向硼中子俘获治疗效应。结果表明，胃癌免疫脂     

七叶皂苷钠对小鼠急性淋巴白血病 L1210 细胞的体内外作用研究

目的 探讨七叶皂苷钠对小鼠急性淋巴白血病 L1210 细胞的体外增殖抑制作用和体内抗白血病作用。 方法 采用 MTT 法观察七叶皂苷钠对小鼠急性淋巴白血病细胞 L1210 的体外增殖抑制作用;建立 L1210 小鼠白血病移植模型,观察七叶皂苷钠对荷瘤小鼠生命的延长率,以及对化疗药阿霉素的增敏作用,并绘制生存曲线。 结果七叶皂苷钠 (20～60 μg/mL) 对 L1210 细胞具有一定的增殖抑制作用,作用呈时间和剂量依赖性,体外作用 72 h 的 IC５０为 (24.86±2.23) μg/mL;体内实验结果显示,七叶皂苷钠中、高剂量组能明显延长荷瘤小鼠的生命期,生命延长率分别为 32.3% 和 25.8% ( P ＜0.01),低剂量组对常规化疗药阿霉素具有增敏作用。 结论 七叶皂苷钠能有效抑制 L1210 细胞体外增殖,对体内 L1210 白血病小鼠移植模型也具有一定的治疗效果。     

白藜芦醇对恶性黑色素瘤生长抑制作用的体外及体内研究

目的:研究白藜芦醇在体外和体内对恶性黑色素瘤的抗肿瘤效果.方法:采用MTT法测定白藜芦醇对小鼠B16及人A375细胞的增殖抑制率;用Western blot方法检测白藜芦醇对B16细胞中p-Akt蛋白表达的影响;建立B16皮下种植瘤小鼠模型,观察不同剂量的白藜芦醇对小鼠皮下瘤的生长抑制作用.结果:体外实验发现在白藜芦醇对B16及A375细胞均显示了良好的浓度依赖性;Western blot显示了不M浓度的白藜芦醇有效的抑制了p-Akt蛋白的表达,提示其抗肿瘤机制可能与此有关;体内实验证实不同浓度的白藜芦醇对恶性黑色素瘤的生长产生了明显的抑制作用.结论:体外和体内实验证实白藜芦醇有效的抑制了恶性黑色素瘤的生长,并且发现其抗肿瘤机制可能通过抑制p-Akt蛋白的表达来实现.这为白藜芦醇在恶性黑色素瘤的临床治疗的可能应用提供了实验基础,也为我国中草药的抗肿瘤效果的开发提供了具体思路.     

The anti-HSV-2 effect of alumen: In vitro and in vivo experimental studies

This study investigated the anti-HSV-2 effect of alumen through in vitro and in vivo expe-riments.Viable cell counting was employed to assess the toxicity of alumen on Vero cells.The inhibition rate of HSV-2 was defined as the cytopathic effect(CPE)of the cells infected with the virus.Alumen suppositories of different concentrations were vaginally applied to the guinea pigs which were then infected with HSV-2 via a vaginal route.The clinical symptoms were observed and the local virus titer calculated.The results showed that alumen had an in vitro anti-HSV-2 effect by means of antiviral dup-lication,direct killing of the virus,and antiviral adsorption.Alumen suppositories of different concen-trations could reduce or completely inhibit HSV-2 infection in guinea pigs.It was concluded that alumen had an in vitro anti-HSV-2 effect through multiple approaches and it could suppress in vivo vaginal HSV-2 infection of guinea pig to some extent.     

紫杉醇纳米复合物体内体外抗宫颈癌活性评价

 目的 利用二硫键共价链接紫杉醇（paclitaxel，PTX）与荧光介孔二氧化硅纳米粒（fluorescent mesoporoussilica nanoparticles，FMSN），形成一种可控药物投递系统（controlled drug delivery system，CDDS），并用体内体外实验评价其抗宫颈癌活性。方法 HeLa细胞分别与PTX、PTX-FMSN和空白对照共培养。细胞IQ工作站和流式细胞仪检测细胞的生长和凋亡情况。血检和组织病理学研究评价药物抗肿瘤作用和毒性作用及不良反应。结果 体外研究表明，PTX-FMSN与PTX溶液相比，对HeLa细胞具有相似的抑制细胞增殖和促凋亡作用。经PTX和PTX-FMSN治疗的荷瘤裸鼠，HeLa细胞坏死率并无明显差异，但在相同剂量下，PTX-FMSN的毒性作用及不良反应明显小于PTX。结论 由于毒性作用及不良反应的降低，这种氧化还原性CDDS具有被应用于宫颈癌临床治疗的美好前景。     

氟罗沙星注射液的体内外抗菌活性研究

采用体外抗菌活性和体内保护作用等试验确定氟罗沙星注射液对临床分离的革兰氏阳性和阴性菌的抗菌活性。氟罗沙星注射液对本试验中的大多数革兰氏阳性和阴性菌均具有良好的抗菌活性;对革兰氏阴性菌和葡萄球菌的抗菌活性强于链球菌。接种量从１０３增加到１０７ＣＦＵ／点时,该药物对试验菌的ＭＩＣ值变化不大;但培养基ｐＨ值为酸性时（ｐＨ＝５）,药物对试验菌的抗菌活性明显降低４～３２倍。体内保护试验表明：氟罗沙星注射液对大肠杆菌感染的小鼠有良好的保护作用;氟罗沙星注射液对四株试验菌的体内抗菌活性与对照药物乳酸环丙沙星注射液相近。     

In vitro and in vivo antibacterial activity of Pogostone

Background Our pervious antibacterial studies on several traditional Chinese medicines have found that Patchouli oil from Pogostemon cablin had significant antibacterial activity against methicillin-resistant Staphylococcus aureus （MRSA）,which has spread worldwide and infected innumerable people.In order to find the more active natural substances in Patchouli oil,one of the major components,Pogostone,was isolated and its antibacterial activity was evaluated in vitro and in vivo in this study.Methods In vitro test,Pogostone was screened for antimicrobial properties against 83 bacteria comprising 35 gram positive and 48 gram negative bacteria strains via the agar double dilution method.In vivo test,specific pathogen free （SPF） strain of both male and female white Kunming mice,weighing 18-22 g,were used to test the protective ability of Pogostone after being injected with the median lethal doses （MLDs） of the tested strains.Results In vitro test,Pogostone could inhibit both gram negative bacteria （0.098-1 600 μg/ml） and gram positive bacteria（0.098-800 μg/ml）.For Corynebacterium xerosis and some Chryseobacterium indologenes,the minimum inhibitory concentration （MIC） values of Pogostone were extremely low （＜0.098 μg/ml）.It was significant that Pogostone was also active against some drug-resistant bacteria like MRSA.Furthermore,Pogostone showed antibacterial activity in vivo against Escherichia coli （E.coli） and MRSA via intraperitoneal injection.Ninety percent of the mice infected with E.coil could be protected at the concentrations of 50 and 100 mg/kg,and 60％ of the mice at 25 mg/kg,while the rate of protection for the mice infected with MRSA was 60％ and 50％ at doses of 100 and 50 mg/kg,respectively.Conclusion Pogostone could be developed as a potential antibacterial agent for clinical therapy.     

In vitro and in vivo study of octacosanol metabolism

BACKGROUND: Policosanol is a mixture of very-long-chain aliphatic alcohols purified from sugar cane wax with cholesterol-lowering effects, whose main component is octacosanol. Scarce data about the metabolism of octacosanol and the other fatty alcohols composing policosanol have been published. METHODS: Human fibroblasts were cultured in presence of (3)H-octacosanol during 0.5, 2 and 4 h. Lipid extracts were analyzed by thin layer chromatography, and the spots corresponding to octacosanol and octacosanoic acid were identified comparing with authentic standards. Spots were scraped, transferred to vials and radioactivity was measured. For corroborating the presence of octacosanol and octacosanoic acid, samples were analyzed by gas chromatography-mass spectrometry (GC-MS). The in vivo study of octacosanol metabolism was conducted in rats and Macaca arctoides monkeys. Rats were orally administered with policosanol (60 mg/kg) and free octacosanol and octacosanoic acid were identified in liver and plasma by GC-MS at various time intervals. Monkeys were orally and endovenously treated with policosanol (10 mg/kg) and the presence of free octacosanol, octacosanoic acid and some chain-shortened FA was investigated. RESULTS: When fibroblasts were cultured in presence of (3)H-octacosanol, three spots were found: a first one corresponded to octacosanoic acid, a second to octacosanol and a third one remained unidentified. The radioactivity on the spot of octacosanoic acid slightly decreased throughout the incubation but increased in the third spot. Octacosanol and free octacosanoic acids were also identified in plasma of monkeys orally administered with policosanol. In addition, plasma samples showed free saturated acids, palmitic acid being the most abundant, followed by oleic and mystiric acids. Unsaturated acids (oleic and palmitoleic) were also observed. CONCLUSIONS: The present study demonstrates that octacosanoic acid is formed after incubation of fibroblast cultures with (3)H-octacosanol and after oral dosing with policosanol to rats. In addition, we demonstrated that shortened saturated (myristic, palmitic and stearic) and unsaturated (oleic, palmitoleic) FA are also formed after oral dosing with policosanol to monkeys. The present results are consistent with the fact that octacosanol metabolism is linked to FA metabolism via beta-oxidation, but further studies need to explore the occurrence of more metabolites proving such hypothesis.     

多西紫杉醇载药纳米微球的制备?表征以及对人卵巢癌SKOV3细胞的体内外抗肿瘤效果评价

目的: 制备可生物降解高分子材料作载体的多西紫杉醇载药纳米微球(Doc-np),检测其各项特征,并考察其体内外抗肿瘤效果?方法:采用溶剂分散法制备Doc-np?用原子力显微镜和透射电子显微镜观察Doc-np的形态,并用动态光散射仪测定纳米微球的粒径?通过紫外分光光度法测定并计算得出Doc-np的载药量和包封率,再测定该微球的体外释放曲线?在体外及体内考察Doc-np对人卵巢肿瘤SKOV3细胞系的杀伤效果?结果:Doc-np为不规则的圆形,表面光滑,其包封率在90%以上?体外释放曲线显示了Doc-np良好的缓释特性,由细胞实验结果看出Doc-np在体外的抑瘤效果更强?体内实验显示通过肿瘤局部的针对性给药,Doc-np与Doc相比明显延缓肿瘤的生长,显示出优于裸药的抗肿瘤效果?结论:Doc-np与Doc相比可以明显抑制人卵巢肿瘤的生长?肿瘤局部给药增强了Doc-np的抗肿瘤效果,作为一种更为有效的给药途径值得进一步研究?     

问号钩端螺旋体胶原酶体内和体外的活性研究

目的通过检测致病性钩端螺旋体（钩体）胶原酶在体内、外的活性,探讨问号钩体黄疸出血群赖型赖株（赖株）假定的胶原酶（LA0872）在钩体病出血中的可能作用。方法在大肠杆菌中克隆、表达重组赖株la0872,并行相关鉴定及蛋白酶学活性检测。比较不同毒力菌株赖株、赖株减毒株和双曲钩体三宝垄群montevalerio型Monte Valerio株钩体胶原酶的基因序列特异性及转录和酶活水平的差异。测定豚鼠和沙鼠赖株感染模型的血清胶原酶活性水平变化。结果成功构建的重组质粒经酶切和测序鉴定显示位点连接正确,插入序列与GenBank公布的la0872序列完全一致,并证实其具有胶原酶活性;不同毒力菌株中的钩体胶原酶基因序列一致,转录和酶活水平均无显著性差异;豚鼠和沙鼠感染赖株后,宿主体内血清胶原酶活性水平与未感染赖株动物比较差异无统计学意义（P〉0.05）。结论首次表达和鉴定了有活性的钩体胶原酶,但其在钩体病出血中的作用尚待证实和探讨。     

司丙红霉素的体内外抗菌活性研究

目的 评价司丙红霉素的体内外抗菌活性。方法 采用琼脂平皿二倍稀释法测定司丙红霉素对800株临床分离致病菌的体外抗菌作用，并与红霉素、罗红霉素、克拉霉素、阿奇霉素、哌拉西林、环丙沙星和头孢噻肟进行比较；采用小鼠腹腔感染模型，观察司丙红霉素口服对金黄色葡萄球菌、化脓链球菌、肺炎链球菌感染小鼠的体内疗效，并与红霉素进行比较。结果 司丙红霉素对112株甲氧西林敏感金黄色葡萄球菌（金葡菌）（MSSA）、36株不产酶金葡菌、111株化脓链球菌、32株肺炎链球菌、40株流感嗜血杆菌和30株副流感嗜血杆菌的抗菌活性较强，MIC50值分别为0．5、4、4、0．125、4、0．125mg／L；司丙红霉素对73株耐甲氧西林金葡菌（MRSA）、149株产酶金葡菌、53株甲氧西林敏感表皮葡萄球菌（表葡菌）（MSSE）、128株耐甲氧西林表葡菌（MRSE）、153株产酶表葡菌、28株不产酶表葡菌、107株粪肠球菌、37株大肠埃希菌、37株铜绿假单胞菌和37株肺炎克雷伯菌的抗菌活性均较弱，MIC50值除肺炎克雷伯菌为32mg／L外，其余均在128mg／L以上。司丙红霉素对金葡菌显示抑菌作用，对化脓链球菌在2～4倍MIC浓度时显示杀菌作用，司丙红霉素抗金葡菌和化脓链球菌的活性随着pH的增加而增强，血清和接种量对司丙红霉素抗金葡菌和化脓链球菌的活性无明显影响。司丙红霉素对金葡菌、化脓链球菌和肺炎链球菌感染小鼠具有很强的保护作用，对化脓链球菌感染小鼠的疗效优于红霉素。结论 司丙红霉素具有较强的体内外抗菌活性，有进一步开发研究价值。     

徐长卿水提物抗乙型肝炎病毒的体外实验研究

目的初步探讨徐长卿水提物的抗乙型肝炎病毒(HBV)作用.方法通过徐长卿水提物作用于体外培养的2.2.15细胞株,观察其对2.2.15细胞株分泌HBsAg和HBeAg的影响,以初步评价其抗HBV作用. 结果徐长卿水提物作用于2.2.15细胞株12d后,对2.2.15细胞株的半数毒性浓度为62.65g/L,对HBsAg的半数抑制浓度小于0.78 g/L、对HBeAg的半数抑制浓度为10.13 g/L;对HBsAg治疗指数大于80.32,对HBeAg治疗指数为6.18.结论徐长卿水提物在体外细胞培养中对两抗原的分泌有较好的抑制作用.     

Antimutagenicity of Propolis Against Some Mutagens in vivo and in vitro

OBJECTIVE: To evaluate the antimutagenicity of propolis in vivo and in vitro. METHODS: Salmonella typhimurium strains TA98 and TA100 were used as a test model in vitro against a direct mutagen DMC and an indirect mutagen 2AF with or without S9 mix, and MN formation of mice bone marrow cell and CAs induction of mice testicle cell were applied as a test model in vivo against two mutagens CP and MMC. RESULTS: The present study clearly demonstrated that propolis could inhibit mutagenicity of both DMC and 2AF directly in a dose-dependent manner, and significant antimutagenic effects (P < 0.05) were obtained in TA98 strain at 2000 and 3000 microg/plate. It also could inhibit mutagenicity of both DMC and 2AF to TA98 strain in a dose-dependent manner, with significant antimutagenic effects (P < 0.05) appeared at 1000, 2000, and 3000 microg/plate. The results of antimutagenicity test in vivo revealed that propolis could inhibit MN formation significantly (P < 0.05) at the doses of 45.0 and 135.0 mg/kg b. w., and decrease the frequency of chromosome aberrants and chromosome aberrant cells significantly (P < 0.05) only at the dose of 135.0 mg/kg b. w. CONCLUSION: The propolis is a good inhibitor for mutagencity of DMC and 2AF in vitro, as well as for CP and MMC in vivo.     

Activity of dihydroartemisinin against Leishmania donovani both in vitro and vivo

Visceral leishmaniasis (VL), also known as kala-azar, is a vector-borne disease caused by protozoa of the Leishmania donovani complex. It is usually fatal if untreated. Pentavalent antimonials have been the mainstay of treatment for more than 60 years, but thesedrugs have serious side effects and progressive antimonialresistance. Other proven therapeutics such as amphotericin B, pentamidine, and paromomycin arecostly without oral formulation and unpleasant side effects. Thus, new chemotherapeutic drugs are required to supplement or replace currently availabledrugs.     

Activity of dihydroartemisinin against Leishmania donovani both in vitro and vivo

Visceral leishmaniasis (VL), also known as kala-azar, is a vector-borne disease caused by protozoa of the Leishmania donovani complex. It is usually fatal if untreated. Pentavalent antimonials have been the mainstay of treatment for more than 60 years, but these drugs have serious side effects and progressive antimonial resistance.1 Other proven therapeutics such as amphotericin B, pentamidine, and paromomycin are costly without oral formulation and unpleasant side effects.2 Thus, new chemotherapeutic drugs are required to supplement or replace currently available drugs.     

散结抗瘤方体内和体外抑瘤作用的实验研究

目的 探讨散结抗瘤方体内和体外抗肿瘤的可能机制。方法体内实验：对荷瘤小鼠（SRS-82瘤株）用散结抗瘤方煎剂灌胃,计算抑瘤率,电镜下观察肿瘤细胞形态,并提取肿瘤细胞DNA做琼脂糖凝胶电泳。体外实验：传代培养SRS-82细胞,用散结抗瘤方含药血清干预细胞后,检测活细胞数、细胞凋亡率、Bcl-2蛋白MMP-2mRNA的表达强度。结果与空白对照组比较,散结抗瘤方组瘤重显著减轻,电镜下可见肿瘤细胞出现凋亡特征性改变;细胞活力明显降低．细胞凋亡率明显升高,细胞Bcl-2蛋白的表达强度降低。结论散结抗瘤方在体内有明显的抑瘤作用。在体外对肿瘤细胞也有明显的抑制增殖作用。该方抑瘤作用可能与其诱导肿瘤细胞凋亡密切相关,机制可能与下调抑制凋亡基因Bcl-2的表达有关。散结抗瘤方对肿瘤细胞MMP-2的表达没有影响。