黄连内生真菌抑乙酰胆碱酯酶活性菌株的筛选及鉴定

目的 从黄连(Coptis chinensis)植株内分离筛选具有较高乙酰胆碱酯酶抑制活性的内生真菌，并对筛选出的强抑制活性菌株进行菌种分类及鉴定，为阿尔茨海默病新药的研发提供潜在的菌种资源。方法 采用改良Ellman法对黄连植株内分离获得的内生真菌菌株进行乙酰胆碱酯酶抑制活性测定，对筛选的活性较强菌株，根据菌株的形态学特征结合分子生物学方法进行菌种鉴定。结果 在黄连根、茎和叶部共分离获得24株内生真菌，有9株对乙酰胆碱酯酶有抑制活性。活性菌株中SCLH12抑制活性较强，经初步鉴定该菌株为枝孢菌属的Cladosporium tenuissimum。结论 从黄连根部筛选分离出乙酰胆碱酯酶抑制活性菌株，其中菌株SCLH12(Cladosporium tenuissimum)具有较强乙酰胆碱酯酶抑制活性，可为开发新的乙酰胆碱酯酶抑制剂(AChEI)奠定基础。     

水仙内生真菌乙酰胆碱酯酶抑制活性菌株的筛选及鉴定

目的 从药用植物水仙中分离、筛选对乙酰胆碱酯酶(AChE)具较高抑制活性的菌株并确定其分类归属.方法 采用组织切块法对水仙内生真菌进行分离纯化,纯化后的菌株发酵培养,用乙酸乙酯萃取发酵液,以改良的Ellman方法进行AChE抑制活性的筛选,对获得的阳性菌株SX-S101进行形态学观察,用分子生物学方法鉴定其分类地位.结果 从水仙的根、鳞茎中共分离到18株内生真菌,活性检测筛选出3株对AChE具较强抑制作用的菌株,其IC50分别为0.4562、0.3897、0.4122 g·L-1;对SX-S101进行分类鉴定,确定其为子囊菌纲(Ascomycetes) 喙枝孢属(Rhinocladiella)菌株Rhinocladiella sp.LA60.结论 首次从水仙内生真菌中筛选出了1株AChE抑制活性较高的菌株Rhinocladiella sp.LA60,为后续开发阿尔采末症治疗药物提供依据.     

具有抗氧化活性的绞股蓝内生真菌的分离及研究

从药用植物绞股蓝根部分离内生真菌,筛选出抗氧化活性菌株并对其成分进行初步检测,鉴定目的菌株。采用常规方法分离绞股蓝根部内生真菌;DPPH法和Fe3+还原力测定抗氧化能力;TLC和HPLC检测内生真菌代谢产物中绞股蓝皂苷;分子生物学方法进行种属鉴定。分离得到的10株内生真菌中G4菌株具有一定的抗氧化活性,经检测发现G4的胞内产物中含有绞股蓝皂苷,鉴定其为柔膜菌目(Helotiales)。     

金荞麦中具抗菌活性内生真菌的筛选和鉴定

目的 研究药用植物金荞麦的抗菌活性内生真菌.方法 采用PDA培养基,从金荞麦的块根、茎、叶中分离到20株内生真菌,鉴定其抗菌活性,并对抗菌活性菌株进行形态学特征和分子鉴定.结果 发现其中1株内生真菌FD12对金黄色葡萄球菌有抗菌活性,但对大肠杆菌、黑曲霉无抑制作用;经形态学特征和分子鉴定,确定菌株FD12为黑肉座菌Hypocrea nigricans.结论 文中结果可为保护金荞麦野生资源,寻找和开发新型抑菌生物制剂开辟了一条新途径.     

红海榄内生真菌BGD22抑菌活性成分的初步研究

 目的：鉴定一株来自于海南文昌红海榄茎的内生真菌菌株BGD22,分离其抑菌活性代谢产物。方法：根据形态学特征和ITS序列分析结果对菌株BGD22进行鉴定;采用各种色谱方法分离乙酸乙酯提取物中的化学成分,并根据化合物的波谱数据确定结构。结果：菌株BGD22被鉴定为黑曲霉的一种,从该菌株中分离到3个化合物：草酸二甲酯（1）、枸橼酸二甲酯（2）和麦角甾醇（3）,其中枸橼酸二甲酯具有较弱的抑制藤黄八叠球菌和枯草芽孢杆菌生长的活性,最低抑菌浓度（MIC）分别为0.5和0.9 mg?mL-1。结论：从红海榄茎中分离到的内生真菌黑曲霉可分泌抑菌活性代谢产物枸橼酸二甲酯。     

具有抗氧化活性的北五味子内生真菌的筛选及鉴定

目的:对具有抗氧化活性的北五味子内生真菌进行筛选及鉴定。方法:利用组织分离法,从野生北五味子的根、茎、叶及果实中分离内生真菌,分别采用1,1-二苯基-2-三硝基苯肼(DPPH)自由基清除法及羟自由基清除法筛选具有抗氧化活性的内生真菌;并对菌株的总DNA进行提取,利用通用引物ITS1和ITS4对菌株18S r DNA ITS序列进行扩增和测序,将测序结果进行同源性比对分析,确定活性菌株的分类地位。结果:从北五味子中共分离得到23株内生真菌。其中,从根部分离出的编号为GSR-12的菌株表现出较强的抗氧化活性,其DPPH自由基清除率和羟自由基清除率分别为87.96%和82.31%;经比对分析后鉴定GSR-12为粉红粘帚霉(Clonostachys rosea)。结论:分离自野生北五味子根部的1株粉红粘帚霉具有较强的抗氧化活性。     

红海榄内生真菌AGR12及其抑菌活性代谢产物

目的 鉴定一株分离自海南文昌红海榄茎的内生真菌菌株AGRl2,分离和鉴定其抑菌活性代谢产物.方法 根据菌株的形态学特征和ITS序列鉴定菌株AGR12.通过硅胶柱层析、Sephadex LH-20凝胶柱层析、制备薄层层析及重结晶等方法分离纯化代谢产物,并根据它们的紫外、质谱、氢谱和碳谱等光谱数据确定化合物结构.结果 菌株AGRl2被鉴定为镰孢霉属木贼镰刀菌(Fusarium equiseti),其在优化后的GMPY培养基中28℃、160r/min振荡培养7d后获得的发酵液具有明显的抑菌活性.从发酵液及菌体中共分离到3个化合物,鉴定为equisetin、epi-equisetin和麦角甾醇(ergosterol),其中equisetin对枯草芽孢杆菌和金黄色葡萄球菌的生长具有明显的抑制活性,最低抑菌浓度(MIC)均为32Bg/mL;epi-equisetin对枯草芽孢杆菌的生长也具有明显的抑制作用,MIC为32μg/mL.结论 木贼镰刀菌菌株AGRl2为首次从红树植物中分离到的内生真菌,其分泌的代谢产物equisetin和epi-equisetin具有明显的抑菌活性.     

大狼毒内生真菌的分离、鉴定与抑菌活性研究

目的 分离和鉴定大狼毒根中的内生真菌,并检测其抑菌活性.方法 用组织块法分离,点植法对分离菌株进行分类鉴定;选择3种人类病原细菌作为指示菌进行体外抑菌试验.结果 分离获得内生真菌41株,鉴定为1纲、4目、6科、15属;其中9株有稳定的抑菌活性,尤其对大肠杆菌有显著抑制作用.结论 大狼毒根中存在丰富的内生真菌,其代谢产物具有一定的抑菌作用,可作为筛选抑菌活性物质的新资源.     

金银花内生真菌的分离鉴定及抑菌活性研究

目的 从药用植物金银花中筛选出对指示菌有抑制作用的内生真菌,并对其抑菌特性进行研究.方法 采用组织块法对金银花内生菌进行分离;用形态学方法对内生真菌进行鉴定;以大肠埃希菌、金黄色葡萄球菌为指示菌,对内生真菌发酵产物进行抑菌试验,并对活性物质的热稳定性进行了研究.结果 分离获得38株内生真菌,鉴定为5目、5科、8个属;其中2株有抑菌活性,且发酵液具有良好的热稳定性.结论 金银花组织中存在丰富的内生真菌,其代谢产物具有一定的抑菌作用.     

丹参内生真菌的分离鉴定及抗血小板凝集活性菌株的筛选

目的:分离鉴定丹参内生真菌,考察其代谢产物粗提物的体外抗二磷酸腺苷(ADP)诱导的血小板凝集活性。方法:对丹参的叶、茎和根中的内生真菌进行分离,采用ADP诱导血小板凝集实验测试丹参内生真菌粗提物的抗血小板凝集活性,并用分子生物学方法对筛选出的活性内生真菌进行鉴定。结果:共分离得到80株内生真菌,发现5株内生真菌的粗提物具有良好的抗血小板凝集的作用。结论:丹参中分离的多种活性内生真菌具有潜在的抗血小板凝集活性,值得进一步研究和开发。     

Safety assessment of poloxamers 101, 105, 108, 122, 123, 124, 181, 182, 183, 184, 185, 188, 212, 215, 217, 231, 234, 235, 237, 238, 282, 284, 288, 331, 333, 334, 335, 338, 401, 402, 403, and 407, poloxamer 105 benzoate, and poloxamer 182 dibenzoate as used in cosmetics

Poloxamers are polyoxyethlyene, polyoxypropylene block polymers. The impurities of commercial grade Poloxamer 188, as an example, include low-molecular-weight substances (aldehydes and both formic and acetic acids), as well as 1,4-dioxane and residual ethylene oxide and propylene oxide. Most Poloxamers function in cosmetics as surfactants, emulsifying agents, cleansing agents, and/or solubilizing agents, and are used in 141 cosmetic products at concentrations from 0.005% to 20%. Poloxamers injected intravenously in animals are rapidly excreted in the urine, with some accumulation in lung, liver, brain, and kidney tissue. In humans, the plasma concentration of Poloxamer 188 (given intravenously) reached a maximum at 1 h, then reached a steady state. Poloxamers generally were ineffective in wound healing, but were effective in reducing postsurgical adhesions in several test systems. Poloxamers can cause hypercholesterolemia and hypertriglyceridemia in animals, but overall, they are relatively nontoxic to animals, with LD(50) values reported from 5 to 34.6 g/kg. Short-term intravenous doses up to 4 g/kg of Poloxamer 108 produced no change in body weights, but did result in diffuse hepatocellular vacuolization, renal tubular dilation in kidneys, and dose-dependent vacuolization of epithelial cells in the proximal convoluted tubules. A short-term inhalation toxicity study of Poloxamer 101 at 97 mg/m(3) identified slight alveolitis after 2 weeks of exposure, which subsided in the 2-week postexposure observation period. A short-term dermal toxicity study of Poloxamer 184 in rabbits at doses up to 1000 mg/kg produced slight erythema and slight intradermal inflammatory response on histological examination, but no dose-dependent body weight, hematology, blood chemistry, or organ weight changes. A 6-month feeding study in rats and dogs of Poloxamer 188 at exposures up to 5% in the diet produced no adverse effects. Likewise, Poloxamer 331 (tested up to 0.5 g/kg day(-1)), Poloxamer 235 (tested up to 1.0 g/kg day(-1)), and Poloxamer 338 (at 0.2 or 1.0 g/kg day(-1)) produced no adverse effects in dogs. Poloxamer 338 (at 5.0 g/kg day(-1)) produced slight transient diarrhea in dogs. Poloxamer 188 at levels up to 7.5% in diet given to rats in a 2-year feeding study produced diarrhea at 5% and 7.5% levels, a small decrease in growth at the 7.5% level, but no change in survival. Doses up to 0.5 mg/kg day(-1) for 2 years using rats produced yellow discoloration of the serum, high serum alkaline phosphatase activity, and elevated serum glutamicpyruvic transaminase and glutamic-oxalacetic transaminase activities. Poloxamers are minimal ocular irritants, but are not dermal irritants or sensitizers in animals. Data on reproductive and developmental toxicity of Poloxamers were not found. An Ames test did not identify any mutagenic activity of Poloxamer 407, with or without metabolic activation. Several studies have suggested anticarcinogenic effects of Poloxamers. Poloxamers appear to increase the sensitivity to anticancer drugs of multidrug-resistant cancer cells. In clinical testing, Poloxamer 188 increased the hydration of feces when used in combination with a bulk laxative treatment. Compared to controls, one study of angioplasty patients receiving Poloxamer 188 found a reduced myocardial infarct size and a reduced incidence of reinfarction, with no evidence of toxicity, but two other studies found no effect. Poloxamer 188 given to patients suffering from sickle cell disease had decreased pain and decreased hospitilization, compared to controls. Clinical tests of dermal irritation and sensitization were uniformly negative. The Cosmetic Ingredient Review (CIR) Expert Panel stressed that the cosmetic industry should continue to use the necessary purification procedures to keep the levels below established limits for ethylene oxide, propylene oxide, and 1,4-dioxane. The Panel did note the absence of reproductive and developmental toxicity data, but, based on molecular weight and solubility, there should be little skin penetration and any penetration of the skin should be slow. Also, the available data demonstrate that Poloxamers that are introduced into the body via routes other than dermal exposure have a rapid clearance from the body, suggesting that there would be no risk of reproductive and/or developmental toxicity. Overall, the available data do not suggest any concern about carcinogenesis. Although there are gaps in knowledge about product use, the overall information available on the types of products in which these ingredients are used, and at what concentration, indicates a pattern of use. Based on these safety test data and the information that the manufacturing process can be controlled to limit unwanted impurities, the Panel concluded that these Poloxamers are safe as used.     

HPLC法测定抗妇炎胶囊中木兰花碱、黄柏碱、药根碱、巴马汀、小檗碱、槐果碱、苦参碱、氧化槐果碱、槐定碱和氧化苦参碱

目的 采用高效液相色谱（HPLC）法同时测定抗妇炎胶囊中木兰花碱、黄柏碱、药根碱、巴马汀、小檗碱、槐果碱、苦参碱、氧化槐果碱、槐定碱和氧化苦参碱10种活性成分。方法 采用InerSustain AQ-C₁₈色谱柱（250 mm×4.6 mm,5 μm）,流动相A：乙腈–无水乙醇（80∶20）,流动相B：0.1%磷酸溶液,梯度洗脱,检测波长220 nm,体积流量1.0 mL/min,柱温30℃,进样量10 μL。结果 木兰花碱、黄柏碱、药根碱、巴马汀、小檗碱、槐果碱、苦参碱、氧化槐果碱、槐定碱和氧化苦参碱分别在2.69～134.50、1.95～97.50、0.63～31.50、0.86～43.00、11.95～597.50、0.59～29.50、6.08～304.00、4.85～242.50、1.66～83.00、19.79～989.50 μg/mL线性关系良好（r≥0.999 3）;平均回收率分别为99.11%、98.23%、96.95%、97.78%、100.02%、97.21%、99.66%、99.52%、98.81%、100.08%,RSD值分别为1.04%、1.23%、1.37%、1.65%、0.70%、1.28%、0.65%、0.81%、1.11%、0.63%。结论 建立的HPLC法可用于抗妇炎胶囊中10种活性成分的测定,作为抗妇炎胶囊质量控制方法。     

Arformoterol: (R,R)-eformoterol, (R,R)-formoterol, arformoterol tartrate, eformoterol-sepracor, formoterol-sepracor, R,R-eformoterol, R,R-formoterol

Sepracor in the US is developing arformoterol [R,R-formoterol], a single isomer form of the beta(2)-adrenoceptor agonist formoterol [eformoterol]. This isomer contains two chiral centres and is being developed as an inhaled preparation for the treatment of respiratory disorders. Sepracor believes that arformoterol has the potential to be a once-daily therapy with a rapid onset of action and a duration of effect exceeding 12 hours. In 1995, Sepracor acquired New England Pharmaceuticals, a manufacturer of metered-dose and dry powder inhalers, for the purpose of preparing formulations of levosalbutamol and arformoterol. Phase II dose-ranging clinical studies of arformoterol as a longer-acting, complementary bronchodilator were completed successfully in the fourth quarter of 2000. Phase III trials of arformoterol began in September 2001. The indications for the drug appeared to be asthma and chronic obstructive pulmonary disease (COPD). However, an update of the pharmaceutical product information on the Sepracor website in September 2003 listed COPD maintenance therapy as the only indication for arformoterol. In October 2002, Sepracor stated that two pivotal phase III studies were ongoing in 1600 patients. Sepracor estimates that its NDA submission for arformoterol, which is projected for the first half of 2004, will include approximately 3000 adult subjects. Sepracor stated in July 2003 that it had completed more than 100 preclinical studies and initiated or completed 15 clinical studies for arformoterol inhalation solution for the treatment of bronchospasm in patients with COPD. In addition, Sepracor stated that the two pivotal phase III studies in 1600 patients were still progressing. In 1995, European patents were granted to Sepracor for the use of arformoterol in the treatment of asthma, and the US patent application was pending.     

欧洲刺柏（Juniper）

活性成分与药理作用欧洲刺柏药用部位是其浆果，具有促水排泄、防腐、抗胃肠胀气和抗风湿作用，还可改善胃功能。用作促水排泄药可增加尿量（水丢失），但不增加钠排泄。成分萜品烯-4-醇可增加肾小球滤过率，但刺激肾。欧洲刺柏浆果对单纯疱疹病毒体外显示抗病毒活性，并具抗真菌活性。动物实验显示，欧洲刺柏浆果提取物具有堕胎、抗生育、抗炎、抗胚胎植入、降血压、升血压和降血糖作用。欧洲刺柏浆果油具有兴奋子宫的活性，以及利尿、胃肠道抗菌和刺激作用，该油对平滑肌有阻止解痉作用。     

Oral Presentations (LDD, LPES, LNM, LPAT, LNT, LPPT, LPM)

Probiotic microorganisms have been shown to provide specific health benefits when consumed as food supplements or as food components. The main problem of such products is the poor survival of the probiotic bacteria in the low pH of gastric fluid. However the use of synthetic excipients for enteric coating to prevent the exposure of microorganisms to gastric fluid is limited in food supplementary industry. Therefore the aim of this study was to develop an enteric coating formulation containing shellac as a natural polymer. Shellac possesses good resistance to gastric juice; the major disadvantage of this polymer is its low solubility in the intestinal fluid [1, 2]. Thus films containing different ratios of shellac and water-soluble polymers (sodium alginate, hydroxypropyl methylcellulose (HPMC) and polyvinylpyrrolidon (PVP)) or plasticizers (glycerol and glyceryl triacetate (GTA)) were prepared in order to analyse the films’ melting temperatures (T_m), the changes in enthalpy (ΔH), their capability of taking up water, and their solubility in different media. The release characteristics of the films were studied by loading pellets with Enterococcus faecium M74 and coating them with formulations containing different amounts of shellac and polymer or plasticized shellac. Using dissolution tests, performed according to USP XXXI paddle method, the resistance of the coatings to simulated gastric fluid (SGF, pH 1.2) and the release of cells in simulated intestinal fluid (SIF, pH 6.8) was investigated.The trials showed that an increasing amount of plasticizer results in a decrease of T_m and ΔH of the films whereat glycerol had a superior plasticization effect to GTA. The compatibility of films made of water-soluble polymers and shellac was also concentration dependent. HPMC and PVP showed superior compatibility with shellac compared to sodium alginate, since films containing shellac and more than 10% [w/w] sodium alginate tended to separate into two phases. In the end five formulations containing shellac and either 5% [w/w] glycerol, 10% [w/w] PVP, 20% [w/w] PVP, 10% [w/w] HPMC, or 5% [w/w] sodium alginate emerged as feasible for enteric coating purposes.