Antifungal activities and action mechanisms of compounds from Tribulus terrestris L.

Antifungal activity of natural products is being studied widely. Saponins are known to be antifungal and antibacterial. We used bioassay-guided fractionation to have isolated eight steroid saponins from Tribulus terrestris L. , which were identified as hecogenin-3-O-beta-D-glucopyranosyl （ 1→4 ）-beta-D-galactopyranoside （ TTS-8）, tigogenin-3-O-beta-D-glucopyranosyl （ 1 →4 ）-beta-D-galactopyranoside （ TTS-9 ）, hecogenin-3-O-beta-D-glucopyranosyl （ 1 → 2 ）-beta-D-glucopyranosyl （ 1 4 ）-beta-D-galactopyranoside （ TTS-10 ）, hecogenin-3-O-beta-D-xylopyranosyl （ 1→3 ）-beta-D-glucopyranosyl （ 1→ 4 ）-beta-D-galactopyranoside （ TTS-11 ）, tigogenin-3-O-beta-D-xylopyranosyl （ 1→ 2 ）-[- beta-D-xylopyranosyl （ 1 → 3 ） ]-beta-D-glucopyranosyl （1→4）-[alpha-L-rhamnopyranosyl （1→2）]-beta-D-galactopyranoside （TTS-12）, 3-O-Ebeta-D-xylopyranosyl （1→2）-[beta-D- xylopyranosyl （1→3）]-beta-D-glucopyranosyl （ 1→4）-[alpha-L-rhamnopyranosyl （ 1→2 ）]-beta-D-galactopyranosy13-26-O-beta- D-glucopyranosyl-22-methoxy-（3beta, 5alpha, 25R）-furostan-3,26-diol （TTS-13）, hecogenin-3-O-beta-D-glucopyranosyl （ 1→ 2）-[beta-D-xylopyranosyl （ 1→ 3 ）]-beta-D-glucopyranosyl （ 1 → 4 ）-beta-D-galactopyranoside （ TTS-14 ）, tigogenin-3-O-beta-D- glucopyranosyl （ 1→ 2 ）-[ beta-D-xylopyranosyl （ 1 → 3 ）]-beta-D-glucopyranosyl （ 1 → 4 ）-beta-D-galaetopyranoside （ TTS-15 ）. The in vitro antifungal activities of the eight saponins against five yeasts, Candida albicans, Candida glabrata, Candida parapsilosis, Candida tropicalis and Cryptococcus neoformans were studied using microbroth dilution assay. In vivo activity of TTS-12 in a Candida albicans vaginal infection model was studied in particular. The results showed that TTS-12 and TTS-15 were very effective against several pathogenic candidal species and Cryptococcus neoformans in vitro. It is noteworthy that TTS-12 and TTS-15 were very active against Candida albicans （MIC50 = 10 and 2.3 microg/ml） and Cryptococcus neoformans （MIC50 = 1.7 and 6. 7 microg/ml）.     

蒺藜总皂苷口腔崩解片的制备

目的筛选蒺藜总皂苷口腔崩解片的处方和制备工艺。方法采用L9（34）正交试验法,以崩解时限为指标,以微晶纤维素、低取代羟丙基纤维素、交联乙烯吡咯烷酮、甘露醇等为辅料,筛选最佳处方和制备工艺。结果优化后蒺藜总皂苷口腔崩解片崩解时间短,崩解时间为25 s左右,口感良好。结论蒺藜总皂苷口腔崩解片达到设计要求,制法简便,崩解时间短,口感良好,服用方便。     

蒺藜总皂甙抑制培养的牛脑微血管内皮细胞表达E选择素

目的：观察蒺藜总皂甙（ＧＳＴＴ）对牛脑微血管内皮细胞（ＢＣＭＥＣ）表达Ｅ选择素的影响。方法：流式细胞仪测定白细胞介素１β（ＩＬ－１β）诱导ＢＣＭＥＣ粘附大鼠血单核细胞（ＭＮＣ）和中性粒细胞（ＮＴＰ）的数目；应用ＥＬＩＳＡ法检测ＢＣＭＥＣ表达Ｅ选择素的量。结果：ＢＣＭＥＣ经ＩＬ－１β诱导后，明显增加细胞表面Ｅ选择素的表达；ＢＣＭＥＣ与白细胞的粘附率亦明显增加，对ＭＮＣ的粘附率从（１２．９±０．４）％增至（４５．７±０．６）％，对ＮＴＰ的粘附率从（１４．７±０．３）％增至（４４．９±０．６）％。抗Ｅ选择素单克隆抗体（ＡＥｍＡｂ）能显著降低ＩＬ－１β的诱导作用。在ＩＬ－１β处理前，用ＧＳＴＴ（０．０１～１．００ｇ／Ｌ）与ＢＣＭＥＣ共孵育，则ＧＳＴＴ可浓度依赖性地抑制ＩＬ－１β的诱导作用，降低Ｅ选择素的表达量。结论：ＧＳＴＴ能抑制ＢＣＭＥＣ表达Ｅ选择素     

一种三唑醇类化合物的体内外抗真菌活性研究

目的:观察三唑醇类化合物SL-130的体内外抗真菌活性. 方法:采用美国临床实验室标准化委员会(NCCLS)1997年公布的M27-A方案,实验用菌株包括白念珠菌及非念珠菌致病真菌,用最低抑菌浓度MIC80表示抗真菌活性.此外利用纸片扩散法、时间-杀菌曲线及小鼠的存活率等方法考察SL-130的抗真菌活性.结果:SL-130具有一定的抗真菌活性,体外明显强于氟康唑,体内与氟康唑基本相当.结论:此三唑醇类化合物结构新颖,体内外的抗真菌活性明显,值得进一步研究.     

不同浓度的中药刺蒺藜对黑素细胞和酪氨酸酶的作用

目的观察中药刺蒺藜对酪氨酸酶和黑素细胞的影响。方法体外培养人正常黑素细胞,以甲基偶氮唑蓝比色法(MTT法)测定不同浓度刺蒺藜作用下黑素细胞的增殖情况,以蘑菇酪氨酸酶多巴速率氧化法测定不同浓度刺蒺藜对酪氨酸酶活性的影响。结果(1)刺蒺藜对黑素细胞具有高浓度激活、低浓度抑制作用,在刺蒺藜浓度为0.5~2.0mg/ml时,其最佳激活浓度为1.5mg/ml,最佳抑制浓度为0.5mg/ml(P<0.05);(2)刺蒺藜对酪氨酸酶亦具有高浓度激活、低浓度抑制作用,在刺蒺藜浓度为1~100mg/ml时,其最佳激活浓度为100mg/ml(P<0.05),最佳抑制浓度为10mg/ml(P<0.05)。结论刺蒺藜对酪氨酸酶和黑素细胞均具有双向调节作用。     

不同浓度的中药刺蒺藜对黑素细胞和酪氨酸酶的作用

OBJECTIVE: To study the effects of the decoction of Tribulus terrestris L on tyrosinase activity and melanogenesis. METHODS: MTT was employed to study the proliferation of human melanocytes cultured in vitro, and tyrosinase activity was estimated by measuring the rate of oxidation of DL-dopa, after the cells were treated with different concentrations of decoction of Tribulus terrestris L. RESULTS: The treatment with the decoction increased the amount of melanin at higher concentrations but act to the reverse effect at lower concentrations, with the best concentrations for promoting and inhibiting the cell growth being 1.5 mg/ml (P<0.05) and 0.5 mg/ml (P<0.05) respectively. At high concentrations the decoction enhanced the tyrosinase activity that was inhibited at low concentrations. The best concentrations for enhancing and inhibiting tyrosinase activity were 100 mg/ml (P<0.05) and 10 mg/ml (P<0.05) respectively. CONCLUSION: Tribulus terrestris L decoction exercises a two-way regulation on the activity of tyrosinase and the proliferation of melanocytes.     

The influence of carbohydrases on the growth of fungal pathogens in vitro and in vivo

Mixtures of mycolytic enzymes from various sources release protoplasts from living fungal tissue under suitable conditions. Such enzyme mixtures obtained from Coprinus comatus (mycolase I), Physarum polycephalum (mycolase II) and Lycoperdon pyriforme (mycolase III) are of low toxicity in mammals when given parenterally and are able to cure experimental systemic fungal infections in mice when administered alone or in conjunction with normally ineffective levels of conventional antimycotic drugs such as amphotericin B. The effect is believed to be due to enzymic degradation of the fungal cell wall either killing the fungus directly or enhancing activity of existing antifungal agents by increasing access to the cell interior.     

In vivo and in vitro Studies on the Antitumor Activities of MCP（Malva crispa L.Powder）^1

Four short-term in vivo and in vitro tests were used to further confirm the antitumor activities of MCP,a vegetable powder,prepared from Malva crispa L.(i)In the H22 heption;5-Fu appeared to have more potent antitumor activities and more harmful effects than MCP.(ii) In the micronucleus(MN)test,MCP significantly decreased MN frequency,(iii) In the cancer cell culture systems.the MCP fat-soluble extract revealed inhibitory effects on the growth and proliferation of the human hepatoma and the gastric cancer cells in a dose-response manner.(iv) In the colony formation test,MCP also altered the morphology of human gastric cancer cells.It was suggested that mcp could be consumed not only by healthy subjects for cancer prevention but also by patients with cancer as supplementary treatment in combination with anticarcionogenic drug such as 5-FU,cyclophosphamide(CP).     

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。     

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。     

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.     

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.     

凤仙花水提物体外和体内的抑菌活性

目的：研究凤仙花水提物的体外、体内抑菌活性,为寻找新的抗菌药物提供药理学参考依据。方法：体外实验分为阴性对照组（A组）、青霉素对照组（B组）、链霉素对照组（C组）和凤仙花水提物的不同浓度(0.01、0.10、1.00和10.00 g/mL)组(D~G组）,浸渍法制备凤仙花水提物,沟槽法和纸片法检测凤仙花水提物对金黄色葡萄球菌、大肠杆菌、白色念珠菌、变形杆菌、铜绿假单胞菌、鲍曼不动杆菌及肺炎克雷伯菌的体外抑菌活性。体内实验分为阴性对照组（A组）和凤仙花不同浓度（0.5、5.0及50.0 g/kg)组(B~D组）,以金黄色葡萄球菌和大肠杆菌作为指示菌种,小鼠灌胃给予凤仙花水提物7 d后,腹腔注射金黄色葡萄球菌和大肠杆菌,通过观察小鼠死亡率衡量凤仙花水提物的体内抑菌活性。结果：体外抑菌实验,当凤仙花浓度高于1.00 g?mL-1时,凤仙花水提物对上述7种致病菌均具有抑制作用,抑菌环直径均明显大于阴性对照组（P<0.05）。体内抑菌实验,小鼠灌胃给予5.0　g?kg-1的凤仙花水提物后,具有抑制体内金黄色葡萄球菌和大肠杆菌的作用,与阴性对照组小鼠死亡率（88.9%）比较,5.0 g?kg-1凤仙花水提物组小鼠死亡率（55.6%）明显下降。结论：凤仙花水提物中含有抑菌活性成分。     

远志属植物的皂苷类成分及其药理作用研究进展

远志属植物在我国多数被药用,具有益智安神、镇咳祛痰之功效,现代药理研究还发现其具有降糖和免疫增强等活性。该属植物主要含有皂苷、酮及寡糖多酯等成分,其中皂苷是该属植物的主要活性成分之一,苷元均为齐墩果烷型三萜,糖的种类有葡萄糖、鼠李糖、木糖、芹糖、半乳糖等。现对该类成分的化学结构、波谱学特征及现代药理研究进行总结,以利于这类植物的进一步研究、开发与利用。     

人参内生真菌的分离及其体外抗真菌、抗肿瘤活性

目的:分离人参内生真菌,并探讨其代谢产物粗提物的体外抗真菌、抗肿瘤活性。方法：首次对5年生园参和15年生移山参内生真菌进行了分离,以稻瘟霉（Pyricularia oryzae P-2b）为筛选模型,采用微量稀释法筛选人参内生真菌体外抗病原真菌活性,MTT法检测其体外抗肿瘤活性。结果：从人参中共计分离得到内生真菌48株,其中16株（33.3%）能完全抑制稻瘟霉活性。16株具有活性的内生真菌中11株（22.9%,11/48）抗白念珠菌（Candida albicans）、新生隐球菌（Cryptococcus neoformans）、红色毛癣菌（Trichophyton rubrum）、薰烟曲霉菌（Aspergillus fumigatus）活性较好;5株（10.4%,5/48）抗胃癌细胞（MKN45）、肠癌细胞（LOVO）、肝癌细胞（HepG2）、白血病细胞（HL-60）活性较好。其中,以园参-25（Yuan-25）抗真菌活性最佳,其抗红色毛癣菌的MIC80为4 mg/L,与阳性对照氟康唑的抗菌活性相当;园参-27（Yuan-27）抗肿瘤活性最优,其抗肿瘤细胞的IC50均与阳性对照阿霉素较接近。结论：分离得到的人参内生真菌具有潜在的抗肿瘤、抗真菌活性,值得进一步研究和开发。     

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     

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.     

383例阴道念珠菌感染菌群鉴定及药敏分析

目的 分析念珠菌性阴道炎的临床分离菌特点及抗念珠菌药物的耐药情况.方法 按NCCLS推荐的M27-A方案(肉汤微量稀释法)检测氟康唑等6种抗真菌药对6种共52株深部致病念珠菌临床分离株在体外的抗真菌活性.结果 383例标本中以白色念珠菌居多(96.09%),其次为光滑念珠菌(2.61%),较少的有热带念珠菌(1.04%)与克柔念珠菌(0.26%).药物敏感试验显示:制霉菌素、两性霉素B的敏感性均在90%以上,耐药率以伊曲康唑为最高(35.8%),其次为氟康唑(17.5%).结论 念珠菌性阴道炎的临床分离菌除了白色念珠菌外,尚有光滑念珠菌、热带念珠菌及克柔念珠菌等菌种,制霉菌素与两性霉素B对念珠菌的敏感性较好,而伊曲康唑与氟康唑的耐药率比较高,应引起临床医师的重视.     

In vitro activity of clindamycin and other antimicrobials against gram-positive bacteria and Hemophilus influenzae.

Summary: Seven antimicrobials--clindamycin, penicillin, ampicillin, cloxacillin, erythromycin, lincomycin and cephalexin--were found to have a high degree of activity in vitro against 256 isolates of gram-positive bacteria and Hemophilus influenzae. Clindamycin was clearly superior against staphylococci and 3.12 mug/ml or less of clindamycin inhibited all 35 isolates of H. influenzae. Synergism was not demonstrated when clindamycin was tested in combination with sulfisoxazole or sulfamethoxazole by either the agar dilution or 24-hour growth curve method. This was true for penicillin as well, and the effect was independent of sulfonamide sensitivity. The erythromycin-sulfonamide combination was synergistic against 6 of 10 strains studied by the growth curve method; this effect was not demonstrable by the agar dilution method.