车前(lantago asiatica L.)毛状根诱导技术研究

为建立发根农杆菌Ri质粒介导的车前(lantago asiatica L.)转化体系,通过发根农杆菌A4和R1601侵染车前无菌苗的外植体诱导毛状根.实验结果表明,车前外植体最佳侵染时间为8 min;两种菌液均可以诱导车前叶片、叶柄和根产生毛状根,车前叶片和叶柄毛状根的诱导率低于40%,车前根部毛状根的诱导率高于80%;经过分子鉴定,Ri质粒T-DNA的rolB基因已成功整合到毛状根中,表明车前的根是适宜毛状根诱导的最佳外植体,可为今后车前毛状根的扩大培养奠定基础.

Anti-fatigue Effect of Water Extract from Centella asiatica(L.)Urban on Mice

[Objectives]To study the anti-fatigue effect of water extract from Centella asiatica(L.)Urban on mice.[Methods]After intragastric administration of low,medium and high concentration of water extract from Centella asiatica(L.)Urban for 14 d,the rotarod time and the content of serum lactic acid in mice were determined,respectively.[Results]Compared with the control group,the rotarod time of mice in the low and medium concentration groups was significantly prolonged(P<0.05),but there was no significant difference between the control group and the high concentration group;the content of serum lactic acid in the medium concentration group was significantly lower than that in the control group(P<0.01),but there was no significant difference between low concentration group and high concentration group and the control group.[Conclusions]A certain concentration of water extract from Centella asiatica(L.)Urban had a good anti-fatigue effect.

Isolation and purification of phenylethanoid glycosides from plant extract of Plantago asiatica by high performance centrifugal partition chromatography

Two phenylethanoid glycosides （PhGs） were isolated and purified from the aerial parts of Plantago asiatica for the first time by high performance centrifugal partition chromatography （HPCPC） using ethyl acetate-n-butanol-ethanol-water （0.5：0.5：0.1：1, v/v/v/v）. A total of 45.6 mg of compound 1 and 293.8 mg of compound 2 were purified from 1341 mg of the n-butanol extract of P. asiatica. The structures of the two PhGs were tentatively identified as plantamajoside and aeteoside or isoacteoside by eleetrospray ionization multi stage tandem mass spectrometry （ESI-MS^n） in the negative ion mode.

Isolation and Purification of Plantamajoside and Acteoside from Plant Extract of Plantago asiatica L. by High Performance Centrifugal Partition Chromatography

Two phenylethanoid glycosides（PhGs）, plantamajoside and acteoside, were isolated and purified from the aerial parts ofPlantago asiatica for the first time by high performance centrifugal partition chromatography（HPCPC） with ethyl acetate, n-butanol, ethanol and water（0.5：0.5：0.1：1, volume ratio） as solvent system. A total of 45.6 mg of plantamajoside and 293.8 mg of acteoside were purified from 1341 mg of the n-butanol extract of P. asiatica, with a purity of 〉93.3% as determined by HPLC. The HPCPC fractions were analyzed by HPLC-DAD and the structures were identified by their retention time, UV, electrospray ionization multi stage tandem mass spectrometry（ESI-MSn） in the negative ion mode, and confirmed by NMR experiments. The characteristic fragment ions of ESI-MS of the two PhGs isolated from Plantago asiatica were discussed, which are specific and useful for the identification of the structures of PhGs.

The Stimulatory Effects of the Antimicrobial Agents Bavistin, Cefotaxime and Kanamycin on <i>In Vitro</i>Plant Regeneration of <i>Centella asiatica</i>(L.)—An Important Antijaundice Medicinal Plant

Antimicrobial agents such as bavistin, cefotaxime and kanamycin were evaluated for their effects on the rapid shoot regeneration from nodal explants of Centella asiatica (L.). Filter sterilized bavistin (250 mg/L) was augmented alone and in combination with cytokinins such as BAP and TDZ into the media to trace the effect on regeneration. On this basis, the potential use of bavistin (150 mg/L) along with BAP (2.0 mg/L) was evaluated which showed the maximum shoot number (6.6) and shoot length (4.4 cm) respectively. Cefotaxime at the concentration of 100 μM/L was found to be effective to obtain the maximum shoot number formation (5.8) with the regeneration frequency (90%). Kanamycin at the concentration of 80 μM/L induced maximum shoot regeneration (5.12). Kanamycin at 100 μM/L or at higher concentrations reduced the shoot regeneration. The best rooting response was noticed when in vitro regenerated microshoots were transferred to the rooting medim which was supplemented with IBA (2.0 mg/L). This combination generates 90% of regeneration frequency and maximum number of roots per shoot (14.2) and high root length (4.2 cm). The rooted plants were acclimatized and transferred to field for survivalance. The addition of antibiotics was found to be more effective and safer for using since their effects on regeneration were found to be negligible.

Biosynthesis of copper nanoparticles supported on manganese dioxide nanoparticles using Centella asiatica L. leaf extract for the efficient catalytic reduction of organic dyes and nitroarenes

In this study we designed a novel,cost‐efficient and green method for the synthesis of copper nanoparticles(Cu NPs)supported on manganese dioxide(MnO2)NPs,using Centella asiatica L.leaf extract as a naturally‐sourced reducing agent,without stabilizers or surfactants.This synthetic process is environmentally‐friendly and avoids the use of toxic reducing agents.Phenolic hydroxyl groups in the leaf extract are believed to reduce Cu2+in solution to generate Cu NPs that are subsequently stabilized on the MnO2NP surfaces.The resulting Cu/MnO2nanocomposite was fully characterized using X‐ray diffraction,transmission electron microscopy,field emission scanning electron microscopy,energy‐dispersive X‐ray spectroscopy and Fourier transform infrared spectroscopy.This material was found to function as a highly active,efficient and recyclable heterogeneous catalyst for the reduction of Congo red,rhodamine B and methylene blue as well as nitro compounds such as2,4‐dinitrophenylhydrazine and4‐nitrophenol in the presence of NaBH4in aqueous media at ambient temperature.The high stability of the Cu/MnO2nanocomposite also allows the catalyst to be separated and reused several times without any significant loss of activity.?2018,Dalian Institute of Chemical Physics,Chinese Academy of Sciences.Published by Elsevier B.V.All rights reserved.

Demographic and behavioral responses of the White-winged Dove to human disturbances during winter in northwestern Mexico

The simultaneous study of demography and behavior might provide a more comprehensive understanding about animal responses to anthropogenic disturbances.The White-winged Dove(Zenaida asiatica)is a frequent dweller of urban and agricultural habitats in North America,Central America,and the Caribbean.Still,research on the species is heavily biased to the USA.We analyzed the response of the species to human disturbances during winter across an urban-agricultural landscape in northwestern Mexico through a demographic and behavioral approach.We modeled the effect of disturbance types(pedestrians,cars,pavement and building cover)on the density of the species.Also,we investigated if surveillance behavior varied across the landscape by estimating and comparing perching proportion among land uses(urban grey areas,urban green areas,cropfields,shrublands).We did not detect the species within cropfields,possibly because food resources were not available for the dove during winter in this land use.Pavement cover limited species’density,as it might reduce the surface of the preferred feeding substrates of the dove.Pedestrians and building cover increased the density of the species,as they might provide food and shelter for the dove,respectively.Surveillance behavior peaked in urban grey areas,whereas it dropped in urban green areas.Differences on threat levels,threat type,and protective cover among land uses might in-fluence such behavioral pattern.Our approach contributes to the knowledge of the species and demonstrates that both demographic and behavioral cues provide complementary evidence for analyzing the impact of human disturbances on animals.

Antifertility properties ofCentella asiatica ethanolic extract as a contraceptive agent: Preliminary study of sperm proteomic

Objective:To investigate the antifertility properties ofCentella asiatica L. ethanolic extract and identify the sperm proteomic changes in treated male rats.Methods: A total of 32 male Sprague-Dawley rats were divided into two groups: control group (n=16) received distilled water and treatment group (n=16) which received 300 mg/kg ofCentella asiatica L. extract for 42 d. After the treatment period, the number of implantation sites was recorded and the sperm proteomic changes were analysed by 2D gel electrophoresis. In addition, the expression of protein spots was quantified by MALDI-TOF analysis.Results:Centella asiatica L. extract resulted in low number of implantation sites in the treatment group (100.00±2.82) compared to the control group (183.00±2.14). The percentage of infertile male rats in the treatment group was higher (43.75%) compared to the control group (18.75%). Proteomic analysis showed the expression of protein spots identified in the treatment group decreased with 234 spots compared to the control group with 282 spots.Conclusions:The results revealed that there was antifertility activity in the male rats with the administration ofCentella asiatica L. ethanol extract and the identified proteins could provide understanding on the adverse effect ofCentella asiaticain male reproductive system.

Vegetative Growth and Molecular Identification of Fusarium equiseti Isolated from Wilt Disease of Centella asiatica L. in Bangladesh

Centella asiatica (L.) is commonly known as Thankuni plant and has ethnobotanical importance in Bangladesh. Present experiment was conceded to investigate the wilt disease of C. asiatica, vegetative growth and molecular characterization of pathogenic fungi. Pathogenic fungus, Fusarium equiseti was identified as a causal agent of wilt disease in C. asiatica. The effect of culture media on the mycelial growth of F. equiseti showed the highest (89.25 mm) on potato dextrose agar (PDA) medium followed by carrot agar (CA) medium and the lowest growth (40.25 mm) was measured in HA medium. The optimal temperature and pH for mycelial growth of F. equiseti were 30&deg;C and 7, respectively. The genetic variation of the selected species of fungi, the internal transcribed spacer (ITS) region was amplified using ITS4 and ITS5 primers and sequenced. The PCR product of the ITS region of F. equiseti was 535 bp. Phylogenetic tree of thirty-seven strains of Fusarium sp. based on the nucleotide sequences of the ITS region using the neighbor-joining method with 1000 bootstrapping indicated that 98% - 100% identity with MN886590.1 JUF0046 (F. equiseti). ITS sequences are generally constant, or show little variation within species, but vary between species in a genus. The ITS region is relatively short and can be easily amplified by PCR using universal single primer pairs. Genetic distance exhibited high level of similarity with identical ITS sequences. To date, no published research articles are found on the molecular identification of F. equiseti, the causal agent of fusarium wilt disease of C. asiatica in Bangladesh.

Induced Mutation for Developing Mutant Rice Lines Tolerant to the Parasitic Weed Striga asiatica (L.) Kuntze

This work aims to screen mutant rice lines tolerant to Striga asiatica.Two rainfed sensitive rice varieties B22 and F154 were used.Plants survival rates of the two parents were significantly lower respectively(9.74a and 11.83a)than those of mutant lines(55.36c to 74.36b);Striga plants emergence/pot were significantly higher for the parents(13.96c and14.89c)compared to the mutants(0.12a to 1.5b);the infection rate of parents(7.37b;7.86b)was higher compared to the mutants(2.27a to 2.74a);fertility rate/plant of parents was lower(20.98%b;22.29%b)but much higher than mutants(72.19%b to 78.35%b);the average panicle number/plant of parents was significantly lower(0.5a;1a)than those of mutants(1.5b to 2.4bc)and the 100 g grain weight of parents are lower(2.35a;2.56a)than those of mutants(3.19b to 3.23b).The culture of those mutant lines may increase rice production and contribute to enhancing food security in Madagascar.

A new triterpene and a saponin from Centella asiatica

A new triterpene and a saponin, named 2α,3β23-trihydroxyurs-20-en-28-oic acid （1） and 2α,3β,23-trihydroxyurs-20-en-28-oic acid O-α-L-rhanmopyranosyl-（1→4）-O-β-D-glucopyranosyl-（1→6）-O-β-D-glucopyranosyl ester （2）, have been isolated from the aerial part of CenteUa asiatica. Their structures were elucidated by spectral methods, including 2D-NMR spectra.

NaCl和NaHCO_3胁迫对车前种子萌发的影响

以车前种子为试材,采用单盐胁迫的方法,研究不同浓度的NaCl和NaHCO3溶液胁迫处理对车前种子萌发及解除胁迫后萌发的影响,探讨车前种子耐盐碱的能力。结果表明：浓度为4‰的NaCl和NaHCO3的溶液均对车前种子的萌发起促进作用,随着浓度的升高,盐胁迫对种子萌发的抑制作用逐渐增强,呈现显著地负相关关系;影响车前种子相对发芽率的NaCl浓度的适宜值和临界值分别是4.69‰和7.42‰;影响车前种子相对发芽率的NaHCO3浓度的适宜值和临界值分别是12.44‰和18.35‰;解除胁迫后,NaCl胁迫的种子复萌率在13.10%~56.33%,NaHCO3胁迫的种子复萌率在12.31%~36.52%。综上所述,车前种子具有较强的耐盐碱能力。

大粒车前子多糖组分PLP-1的结构表征

研究了由大粒车前子多糖分离纯化所得组分PLP-1的一级精细结构特征.通过单糖组成分析、部分酸水解及甲基化分析,并结合NMR,GC和GC-MS等技术测定了PLP-1的一级结构.结果表明,PLP-1由Ara(33.98%),Xyl(58.23%),Rha(2.36%),Glc(2.23%),Gal(2.36%)和Man(0.83%)组成,糖醛酸以GlcA形式存在.PLP-1中主要糖残基有α-T-linked Araf(9.58%),β-T-linked Xylp(8.71%),α-1,3-linked Araf(18.22%),β-1,3-linked Xylp(8.05%),β-1,4-linked Xylp(5.85%),β-1,2,4-linked Xylp(16.98%)和β-1,3,4-linked Xylp(27.52%),GlcA以α-T-linked GlcAp形式存在;此外还有1,2-linked Rhap,T-linked Glcp,1,6-linked Glcp,T-linked Galp,1,3-linked Galp和1,3,6-linked Galp等少量其它残基.根据所得结果推断出PLP-1可能的一级结构.

中药车前子对小鼠气囊滑膜炎细胞因子TNF-α及IL-12影响的实验研究

目的:研究中药车前子对小鼠滑膜炎的抗炎机制。方法:使用T.Mikami's方法用角叉菜胶给小鼠造模,应用双抗体夹心酶联免疫吸附试验法检测灌洗液中TNF-α、IL-12的含量变化。结果:中药车前子组TNF-α、IL-12含量低于对照组,有显著性差异(P<0.01)。结论:车前子能通过抑制滑膜炎症中TNF-α,IL-12的含量进行抗炎。

透骨草提取物诱导人宫颈癌Hela细胞凋亡及其相关机制研究

目的:探讨透骨草提取物诱导人宫颈癌Hela细胞凋亡及其相关机制。方法:荧光显微镜观察Hela细胞形态,透射电镜观察Hela细胞超微结构,流式细胞术检测Hela细胞凋亡率,Western blot检测Bcl-2蛋白和Caspase-3蛋白表达水平。结果:100μg/ml透骨草提取物处理的Hela细胞后,荧光显微镜显示Hela细胞皱缩,细胞核浓缩呈新月状边集。透射电镜可见Hella细胞表面突起和微绒毛减少,核断裂、染色质凝聚且边缘化,有"出芽"现象。流式细胞术显示透骨草提取物处理的Hela细胞凋亡率为(25.90±1.13)%,明显高于对照组(P<0.01)。Western blot结果显示透骨草提取物处理的Hela细胞Bcl-2蛋白和Caspase-3蛋白表达达明显低于与对照组(P<0.05)。结论:透骨草提取物可诱导Hela细胞凋亡,其机制可能与下调Bcl-2蛋白和Caspase-3蛋白表达有关。

车前属(Plantago L.)3种车前种子的生物学比较研究

本研究对黑龙江省车前科(Plantaignaceae)车前属(Plantago L.)的3种野生车前种子的生物学特性进行比较,为黑龙江地产药材车前属植物的引种驯化和开发利用提供依据。采用显微观察、生物学对比等方法,对种子形态和种子萌发的影响因素进行比较研究。结果表明:3种车前的种子形态存在差异,平车前的种子不存在休眠现象,通过低温和赤霉素协同处理有利于大车前和车前种子的萌发。因此,大车前和车前的种子在引种驯化前需要经过低温处理。

合肥城市群:“双C+l线+三角形”的城市体系发展对策分析

合肥在发展城市群的过程中具有优越条件,可以突破行政区划构建跨区域城市组团,成为"上海—南京"城市链的延伸部分,构成"上海—南京—合肥"城市链。合肥城市群在构建过程中需要依托现有基础,形成"双C+l"的城市体系,"双C"即由桐城、六安构成的C西环和由滁州、马鞍山、芜湖构成的C东环;"l"即"淮南—合肥—铜陵"构成的南北方向的城市链,将凤台、长丰、肥东、肥西、巢湖、庐江、铜陵县等整合在一起,以合肥为中心向南北两个方向发展。同时,还要强化构建"合肥—南京—芜湖"三角形,从根本上解决次级核心城市不足、聚集程度差异大以及城市走廊发展不充分等问题。

积雪草药材HPLC指纹图谱的研究

目的:研究积雪草药材的高效液相色谱指纹图谱,为科学评价及有效控制药材质量提供可靠的方法。方法:利用高效液相色谱法,梯度洗脱,测定了16个不同产地的积雪草药材样品。色谱柱为Alltech C18柱(250 mm×4.6 mm5,μm),流动相为乙腈-水,流速为1.0 mL/min,检测波长为205 nm。结果:建立了积雪草药材的HPLC指纹图谱,并检测了16批积雪草药材样品,标定了15个共有峰,通过与对照品的保留时间比对及LC-ESI-MSn进行结构解析,其中5～10号峰分别鉴定为羟基积雪草苷、积雪草苷、槲皮素、山萘酚、羟基积雪草酸、积雪草酸。经相似度计算,各产地药材之间相似性较差。结论:该方法准确可靠,重现性好,可作为控制积雪草药材内在质量的标准。

HPLC-ELSD法测定积雪草中积雪草苷及羟基积雪草苷的含量

积雪草提取物通常被用作化妆品原料,有美白、抗氧化等功效,发挥此功效的主要物质有积雪草苷、羟基积雪草苷。采用高效液相色谱-蒸发光散射检测(HPLC-ELSD)技术,建立了测定积雪草中积雪草苷和羟基积雪草苷含量的方法。选用Unitary C18色谱柱(4.6×250 mm,5μm),以乙腈-水为流动相,梯度洗脱,流速1 mL/min,柱温为30℃,采用Agilent 1260 InfinityⅡ蒸发光散射检测器(漂移管温度70℃,载气流速1.6 L/min)。研究表明,积雪草苷和羟基积雪草苷质量浓度分别在100~702 mg/L,105~738 mg/L范围内,峰面积与质量浓度线性关系良好,相关系数分别为0.9996和0.9997。积雪草苷和羟基积雪草苷平均加样回收率分别为101.20%,98.30%。相对标准偏差(RSD)均在3%以内。方法简单,准确度高,分离效果好,干扰少,可用于积雪草中积雪草苷和羟基积雪草苷的含量测定。

基于UPLC-Q-TOF/MS法分析车前子生品和盐炙品化学成分研究

目的:利用超高效液相串联四级杆飞行时间质谱(UPLC-Q-TOF/MS)法分析车前子及其盐炙品的成分,比较盐炙前后药物化学成分种类和含量的变化,探究车前子炮制前后药效变化的物质基础。方法:ACQUITY UPLC BEH C_(18)色谱柱;流动相:0.1%乙酸水-甲醇体系梯度洗脱;ESI离子源;负离子模式;基于Markerlynx分析软件,利用主成分分析法(PCA)和正交偏最小二乘判别法(OPLS-DA)分析车前子盐炙前后指纹图谱的差异。结果:盐炙品中京尼平苷酸、毛蕊花糖苷、异毛蕊花糖苷的含量相对生品高。京尼平苷酸和毛蕊花糖苷可作为化学标记物区分生品与盐炙品。结论:基于UPLC-Q-TOF/MS法建立了车前子炮制前后的化学成分分析方法。车前子盐炙后有效成分含量增加。本实验的研究成果对于研究车前子盐炙的炮制原理具有重要意义,同时也为车前子盐炙品药效物质基础的阐明提供了重要依据。