A comparison of in vitro anticancerous activity and mechanism of ethanolic extracts from different Ganoderma genus

Five ethanolic extracts from the mycelia of Ganoderma lucidum,G.tsugae,G.oerstedii,G.subamboinense,and G.resinaceum were respectively studied on their anticancerous activities against leukemic HL-60 cell line in vitro.Results showed that all five extracts potently inhibited HL-60 proliferation.The extract from G.lucidum mycelia exerted the highest activity.Annexin V/PI bivariate flow cytometric analysis further revealed that the five extracts significantly induced early apoptosis in HL-60 cells.The results illustrate that not only G.lucidum but also other Ganoderma species can inhibit cancer cells,and their mechanisms are related to induction of apoptosis.     

23种中草药的体外抗菌活性筛选研究

为考察中草药抗菌物质基础筛选出活性提取物,该研究通过80%乙醇冷浸和95%乙醇回流提取制备23种中草药的提取物,采用琼脂扩散法测量抑菌圈直径,用微量液体培养基倍比稀释法测定最低抑菌浓度(minimum inhibitory concentration,MIC)和最低杀菌浓度(minimum bactericidal/fungicidal concentration,MBC/MFC),并测定了提取物对临床4种常见病原菌体外抗菌活性。结果表明:紫珠草、千斤拔、黄龙尾等9种中草药对金黄色葡萄球菌有较强的抑菌活性,其MIC/MBC值除个别菌是12.5 mg·mL~(-1)外,其他都在0.09～3.12 mg·mL~(-1)之间;千斤拔、大红袍、过江龙等5种中草药对铜绿假单胞菌有较强抑菌活性,其MIC/MBC值在3.12～12.5 mg·mL~(-1)之间;紫珠草、千里光、石楠等13种中草药对大肠埃希菌有较强的抑菌活性,其MIC/MBC值在0.09～6.25 mg·mL~(-1)之间;八角对白色念珠菌有较强抑菌活性,其MIC/MFC值在0.78～12.5 mg·mL~(-1)之间。23种中草药的抗细菌活性较好,尤其是千斤拔、大红袍、过江龙、八角、黄药子对金黄色葡萄球菌、大肠埃希菌、铜绿假单胞菌都具有较好的抑菌活性,具有广谱抗菌活性;但对真菌抑菌效果不明显,仅有八角对白色念珠菌有抑菌活性。此外,提取溶剂浓度、提取温度和提取时间对中草药的提取率和活性均有影响,冷提稍优于热提。     

灵芝真菌液体发酵及其产物应用的研究进展

灵芝作为一种白腐真菌,同时也是珍稀的食药用真菌,富含多种生物活性成分。液体发酵技术生产周期短、效率高、产量大、品质稳定,是开发利用灵芝资源的重要途径。近年来,灵芝属真菌菌丝体液体发酵技术的开发与应用取得了较大进展。本文对灵芝属真菌液体发酵产物的主要活性成分及其药用效果、液体发酵工艺优化和发酵产物的应用进行综述,并对本领域的未来进行展望。     

广西五种绞股蓝属植物离体快繁与种质保存研究

以绞股蓝属植物的带芽茎段为材料,研究不同6-BA浓度与NAA 0.02mg·L-1组合对其诱导、分化和增殖的影响,并建立离体快繁体系。结果表明:MS+6-BA 2.0mg·L-1+NAA 0.02mg·L-1最适宜初代诱导,MS+6-BA 2.0mg·L-1+NAA 0.02mg·L-1最适合扁果绞股蓝的增殖培养,而MS+6-BA 1.5mg·L-1+NAA0.02mg·L-1是其它四种植物增殖的最佳培养基,在1/2MS+NAA 1.0mg·L-1上的生根率均达100%。1/2MS与蔗糖40g·L-1对五种植物的保存效果均最好;添加生长抑制剂能有效减缓生长速度,最佳生长抑制剂为ABA和CCC,浓度均为1.0mg·L-1,其中CCC能适合多个物种,连续保存360d的存活率均在94.5%以上;PP333不适合五种植物的保存。活力检测表明,各种质经保存后增殖、生根能力均未下降。     

The effect of blueberry extracts on Giardia duodenalis viability and spontaneous excystation of Cryptosporidium parvum oocysts, in vitro

The protozoan parasites Giardia duodenalis and Cryptosporidium parvum are common causes of diarrhoea, worldwide. Effective drug treatment is available for G. duodenalis, but with anecdotal evidence of resistance or reduced compliance. There is no effective specific chemotherapeutic intervention for Cryptosporidium. Recently, there has been renewed interest in the antimicrobial properties of berries and their phenolic compounds but little work has been done on their antiparasitic actions. The effect of various preparations of blueberry (Vaccinium myrtillus) extract on G. duodenalis trophozoites and C. parvum oocysts were investigated. Pressed blueberry extract, a polyphenolic-rich blueberry extract, and a commercially produced blueberry drink (Bouvrage) all demonstrated antigiardial activity. The polyphenol-rich blueberry extract reduced trophozoite viability in a dose dependent manner. At 167 μg ml⁻¹, this extract performed as well as all dilutions of pressed blueberry extract and the Bouvrage beverage (9.6 ± 2.8% live trophozoites remaining after 24 h incubation). The lowest dilution of blueberry extract tested (12.5% v/v) contained >167 μg ml⁻¹ of polyphenolic compounds suggesting that polyphenols are responsible for the reduced survival of G. duodenalis trophozoites. The pressed blueberry extract, Bouvrage beverage and the polyphenolic-rich blueberry extract increased the spontaneous excystation of C. parvum oocysts at 37 °C, compared to controls, but only at a dilution of 50% Bouvrage beverage, equivalent to 213 μg ml⁻¹ gallic acid equivalents in the polyphenolic-rich blueberry extract. Above this level, spontaneous excystation is decreased. We conclude that water soluble extracts of blueberries can kill G. duodenalis trophozoites and modify the morphology of G. duodenalis and C. parvum.     

狭叶薰衣草的离体培养及挥发性成分的分析

为建立新疆狭叶薰衣草(Lavandula angustifolia)的快速繁殖体系,以种子、茎、叶为外植体,对种子萌发、愈伤组织诱导、丛芽分化和生根的最适培养条件进行了研究;用水蒸气蒸馏法提取狭叶薰衣草挥发油,采用气相色谱-质谱法测定挥发油成分。结果表明,种子浸泡的适宜时间为6 h,切开种皮培养,出芽时间最少为6 d;诱导种子出芽的适宜培养基为MS+6-BA2 mg/L;以茎为外植体诱导愈伤组织效果较好,适宜培养基为MS+6-BA 2 mg/L+2,4-D 1 mg/L;诱导分化丛芽的适宜培养基为MS+6-BA 1 mg/L+NAA 0.5 mg/L;生根的适宜培养基为1/2MS+NAA 1 mg/L+6-BA 0.5 mg/L;盆栽薰衣草和无菌苗薰衣草的挥发油主要成分相差较大,离体培养的薰衣草的主要挥发性成分有叶绿醇、丁香油烃、氧化石竹烯等。     

冷杉侧耳多糖提取工艺优化及体外抗氧化活性

【背景】冷杉侧耳（Pleurotus abieticola）是一种新型食药用真菌,具有潜在的经济价值。【目的】更好地开发利用冷杉侧耳中的多糖成分,挖掘潜在的功能性食品。【方法】采用响应面分析法对冷杉侧耳多糖的提取工艺进行优化,并通过设置不同浓度梯度的多糖和Vc （阳性对照）考察该多糖的1,1-二苯基-2-苦基肼自由基（1,1-diphenyl-2-picrylhydrazyl free radical,DPPH）、2,2''-联氮-双-3-乙基苯并噻唑林-6-磺硫（2,2''-azinobis-3-ethylbenzthiazoline-6-sulphonate,ABTS）和羟基自由基（hydroxyl radical,·OH）的清除能力及铁离子还原能力来评价冷杉侧耳多糖的体外抗氧化活性。【结果】最佳提取工艺：提取时间122.62 min,提取温度60.27℃,液料比28.87：1（体积质量比）,在该条件下,冷杉侧耳纯多糖提取率达到极大值2.39%。该多糖在浓度为1 mg/mL时,DPPH自由基清除率为49.42%,ABTS自由基清除率为88.37%,羟基自由基清除率为19.87%,铁离子还原力为0.273。【结论】本研究为更高效地开发与利用冷杉侧耳多糖提供了理论依据,为进一步挖掘其生物活性及多糖产品打下了基础。     

灵芝液态发酵胞内外多糖结构特征及其活性研究进展

灵芝是一味传统的中药材,具有很高的药用价值,多糖是其主要的活性成分之一,可从其子实体、孢子粉、发酵菌丝体和胞外液中获得。近年来,从灵芝菌丝体和胞外液中发现的多糖越来越得到研究者们的关注。本文从发酵培养基成分及发酵条件对灵芝胞内外多糖的影响、灵芝胞内外多糖的结构、灵芝胞内外多糖生物活性3个方面进行综述,为发酵来源灵芝多糖的开发利用提供科学理论依据。     

蒲公英糖蛋白体内外抗氧化作用研究

为了探讨蒲公英糖蛋白(TMGP)体内外抗氧化活性,采用H2O2、Fe2+、DPPH.和总抗氧化能力反应体系,检测蒲公英糖蛋白体外抗氧化活性,并与VC进行比较;同时建立D-半乳糖所致衰老小鼠模型,测定并比较TMGP组与模型组的血清、肝脏和脑内MDA含量和SOD、CAT、GSH-Px活性。结果显示:(1)随着TMGP浓度增加,其Fe2+、H2O2和DPPH.清除率均有所增加,当TMGP浓度为1.72mg/mL时,对H2O2、Fe2+清除率分别达到76.8%和55%,接近VC水平;当TMGP浓度为1.5mg/mL时,对DPPH.的清除率达到77%,也与VC水平相当,说明TMGP具有一定的体外总体抗氧化能力。(2)将TMGP按低、中、高剂量分别给小白鼠灌胃,可显著增强小鼠体内血清、肝脏、脑组织中SOD、CAT、GSH-Px活性,降低MDA含量,从而提高衰老小白鼠体内的抗氧化能力。研究表明,TMGP具有明显的体内外抗氧化作用。     

Schistosoma mansoni: Antischistosomal activity of the four optical isomers and the two racemates of mefloquine on schistosomula and adult worms in vitro and in vivo

Recent studies have shown that mefloquine (MQ) reveals interesting antischistosomal properties. We examined the antischistosomal activities of the erythro and threo isomers and racemates of MQ on newly transformed schistosomula (NTS) and adult Schistosoma mansoni in vitro and in mice harbouring adult S. mansoni. The in vitro effects in the presence and absence of haemin were monitored by means of microcalorimetry, scanning electron microscopy and phenotypic evaluation. Incubation of NTS with the erythro derivatives at concentrations of 3 μg/ml and above resulted in convulsions, granularity, decrease in heat flow, and death while NTS incubated with the threo derivatives were only affected at high concentrations (100 μg/ml). Extensive tegumental alterations, decrease in metabolic activity, viability, and death were observed when adult schistosomes had been exposed to 10 μg/ml of the erythro compounds. Moderate tegumental and viability changes but reduced heat production rates were observed with the threo derivatives at 10 μg/ml. In the presence of haemin, all MQ derivatives showed pronounced antischistosomal properties against adult S. mansoni in vitro. In vivo, MQ derivatives achieved statistically significant total and female worm burden reductions ranging between 65.4% and 100%. The highest total worm burden reductions of 93.4% and 90.2% were observed following treatment with the erythro and threo racemates, respectively. In conclusion, the optical isomers and racemates of MQ show only moderate stereoselectivity, in particular in vivo. Our results may enhance our understanding of the mechanism of action and therapeutic profile of MQ derivates on schistosomes.     

不同光质对白及组培苗生长及光合特性的影响

白及的自然繁殖率极低,组培育苗是其种苗繁殖的主要方式之一.为探索提高白及组培育苗质量及缩短育苗周期的高效人工光环境,该文以紫花白及(Bletilla striata)为试验材料,研究LED光质对白及组培苗的生长和光合特性的影响.结果表明:提高红蓝光组合中的蓝光占比,有利于促进白及组培苗的生长和生物量的积累,而白及的球茎...     

六角果鸢尾离体快繁技术体系的构建

为构建六角果鸢尾(Iris hexagona)的离体快繁体系,以其幼嫩根状茎为外植体,研究了培养基和植物生长调节剂对不定芽诱导、增殖和植株生根的影响。结果表明,根状茎用0.1%Hg Cl2消毒13 min的效果较佳;不定芽诱导最适培养基为MS+6-BA 1.5 mg L–1+NAA 0.5 mg L–1+蔗糖30 g L–1+琼脂7.5 g L–1,不定芽增殖的适宜培养基为MS+6-BA 0.5 mg L–1+NAA0.2 mg L–1+KT 0.3 mg L–1+蔗糖30 g L–1+琼脂7.5 g L–1;在MS+IBA 1.5 mg L–1+蔗糖30 g L–1+琼脂7.5 g L–1培养基上不定芽生根率可达100%;腐殖土和珍珠岩+泥炭土+蛭石(1∶2∶1)均可作为组培苗移栽的适宜基质,移栽成活率可达100%。     

猴面包树的离体快繁条件筛选

该文以猴面包树(Adansonia digitata)种子为外植体,首先筛选合适的种子预处理及消毒方法,然后经过启动培养获得无菌外植体后在增殖培养基中进行丛生芽诱导,将丛生芽切成单株进行生根壮苗培养,最终建立猴面包树离体快繁技术体系.结果表明:75％酒精浸泡3 min+0.1％升汞消毒15 min消毒效果较佳,污染率为...     

单增李斯特菌胎盘感染的体内外模型研究进展

单核细胞增生李斯特氏菌（Listeria monocytogenes）是重要的食源性致病菌,能引发人类的李斯特菌病,是全球公共卫生问题之一。该菌易感染孕妇,引起胎儿和新生儿的侵袭性李斯特菌病,严重威胁母婴健康。因此,建立有效的单增李斯特菌感染胎盘体内外模型,解析和探究单增李斯特菌经胎盘感染机制,是预防和控制单增李斯特菌感染母婴的关键所在。本文综述了可用于研究单增李斯特菌母婴感染的体内外胎盘模型,总结和讨论了各类模型的优势和局限性;并着重分析了体外三维胎盘屏障模型在单增李斯特菌感染方面的研究进展和未来研究方向。以期为深入解析该菌经胎盘感染的途径、发病机制提供支持,并为预防和控制母婴李斯特菌病提供科学参考。     

Schistosoma mansoni: Schistosomicidal effect of mefloquine and primaquine in vitro

We investigated the effects of the anti-malarials mefloquine and primaquine against the juvenile and adult life stages of Schistosoma mansoniin vitro. Cercariae were incubated with 0.5 μg/ml, 1 μg/ml and 2 μg/ml mefloquine or primaquine and with 1 μg/ml praziquantel for 12 h. Schistosomula, pre-adults and adults were incubated with 0.5 μg/ml, 1 μg/ml and 2 μg/ml mefloquine or primaquine and with 1 μg/ml praziquantel for 7 days. The viability status was classified as viable, damaged or dead and was checked every 3 h for cercariae and every 12 h for schistosomula, pre-adults and adults. Both, mefloquine and primaquine show time and dose-dependent schistosomicidal effects on the four life stages of S. mansoni. The promising in vitro effects on all stages of the blood fluke S. mansoni warrants further evaluation of both anti-malarials and their derivatives for their prophylactic and therapeutic values in early and late schistosomiasis in field trials.     

Effect of enzyme extracts isolated from white-rot fungi on chemical composition and in vitro digestibility of wheat straw

A series of in vitro experiments were completed to evaluate the potential of enzyme extracts, obtained from the white-rot fungi Trametes versicolor (TV1, TV2), Bjerkandera adusta (BA) and Fomes fomentarius (FF), to increase degradation of cell wall components of wheat straw. The studies were conducted as a completely randomized design and analysed using one-way ANOVA. Enzyme activities of the extracts, previously obtained from a liquid culture medium, were characterized in terms of laccase and peroxidase for ligninolytic activity. Carboxymethyl cellulase (CMCase) and avicell digesting cellulase (Avicelase) were used for cellulolytic enzyme assays. Wheat straw samples were incubated with enzyme extracts in a citrate buffer (pH 5.0) in a forced air oven at 25 °C for 6 days. In vitro NDF digestibility (IVNDFD), and the rate and extent of NDF fermentation, without and after incubation with the white-rot enzyme extracts, were determined using a gravimetric microbiological method and a gas production technique, respectively. Results from cell wall chemical composition showed that TV2 and BA enzyme extracts decreased NDF concentration (P<0.05) and that TV1 had higher activity (P<0.05) towards cellulose. There was an increase in IVNDFD (P<0.05), resulting from treatment of wheat straw with enzyme extracts from BA, TV1 and TV2, reaching a difference of 13% for TV2 (P<0.05), versus the non-treated straw control. Treatment with enzyme extract from TV2 caused increased gas production (P<0.05) after the first 20 h of incubation, and also increased the maximum rate of gas production, thus enhancing fermentation kinetics. This study indicates that enzyme extracts from white-rot fungi can be used to develop new approaches to overcome low digestibility of some plant cell walls. Utilization of different substrates to produce enzyme extracts can lead to production of viable ligninolytic complexes which could improve the nutritive value of fibrous feeds.     

CaXMT和TCS1突变体基因串联共表达及体外酶活检测分析

咖啡碱和可可碱是茶叶生物碱的主要组分,且咖啡碱是茶叶重要的滋味物质,随着咖啡碱在食品和药物领域的应用愈发广泛,咖啡碱的生物合成成为新的研究热点.目前市场上的咖啡碱主要靠化学合成,为了探索其生物合成途径,该研究将咖啡黄嘌呤核苷甲基转移酶(coffee xanthosine methyltransferase,CaXMT)基因和茶树咖啡碱合成酶(tea caffeine synthase,TCS1)基因的4个突变体分别串联至同一大肠杆菌表达载体pMAL-c5X,诱导融合蛋白共表达,并进行SDS-PAGE凝胶电泳分析.结果表明:目的蛋白成功表达后,应用超声破碎法制备含有目的蛋白的粗酶液,添加底物黄嘌呤核苷(xanthosine,XR)和甲基供体S-腺苷甲硫氨酸(S-adenosyl-L-methionine,SAM)进行体外酶促反应,将反应产物进行高效液相色谱检测.检测结果显示,pMAL-CaXMT-TM2/3/4的体外酶促反应产物仅有可可碱生成,均未见咖啡碱生成.该研究结果为构建生物合成咖啡碱和可可碱的串联共表达载体奠定了基础,也为进一步研究生物合成咖啡碱和可可碱提供了新思路.     

黄芩苷对嗜水气单胞菌生物膜形成的影响及体内外的抑菌作用

[目的]探讨中药单体黄芩苷对嗜水气单胞菌在体内外生长及生物膜形成的影响.[方法]体外实验中,利用牛津杯法检测抑菌圈直径,结晶紫法检测生物膜的形成,通过泳动实验检测黄芩苷对嗜水气单胞菌运动性的影响,紫外吸收法检测细胞膜完整性,用透射电镜技术观察黄芩苷对细菌形态的影响.体内实验利用草鱼为对象检测黄芩苷对嗜水气单胞菌增殖的影...     

多糖类益生元的筛选及凝结芽孢杆菌体外发酵特性北大核心

【目的】探究9种多糖对凝结芽孢杆菌(Bacillus coagulans)的增殖、产酶特性的影响。【方法】将凝结芽孢杆菌分别添加至菊粉多糖(inulin polysaccharide)、刺五加多糖(Eleutherococcus senticosus polysaccharide)、壳寡糖(chitosan oligosaccharide)、防风多糖(Saposhnikovia divaricata polysaccharide)、低聚木糖(xylo-oligosaccharide)、黄芪多糖(Astragalus polysaccharide)、甘露糖(D-mannose)、白术多糖(Atractylodes macrocephala Koidz polysaccharide)和玉屏风多糖(Yu Ping Feng polysaccharide)为唯一碳源的培养基中,通过菌株生长、酶活性及其体外厌氧发酵等作为指标,筛选出最优益生元。【结果】凝结芽孢杆菌能很好地利用防风多糖、黄芪多糖、白术多糖和玉屏风多糖;添加量为4%的防风多糖和白术多糖,pH差值差异最大,蛋白酶活性差异显著(P<0.05)。体外发酵乳酸活性和总蛋白酶活性均提高,4%白术多糖的乳酸和总蛋白酶活性差异显著(P<0.05);肠道内容物发酵液16S rRNA基因高通量测序结果表明,与对照组比较,添加黄芪多糖、防风多糖、甘露糖3种益生元发酵凝结芽孢杆菌显著降低了气单胞菌(Aeromonas)、α-变形菌(α-Proteobacteria)、链球菌(Streptococcus)、志贺氏杆菌属(Shigella)等致病菌的相对丰度,提高了乳杆菌(Lactobacillus)、厚壁菌门(Firmicutes)、乳酸乳球菌(Lactococcus lactis)、产酸杆菌(Acidobacteria)的相对丰度。【结论】凝结芽孢杆菌发酵4%白术多糖具有较好的产酶性能与益生特性,二者协同发酵添加至饲料中具有较好的发展潜力。