花菊盆炎颗粒提取工艺研究

目的:确定花菊盆炎颗粒最佳提取工艺。方法:应用HPLC测定槲皮素含量,并与正丁醇浸出物总量的综合评分为考察指标,采用正交试验法L 9(34)优选提取工艺条件。结果:最佳提取工艺为处方中头花蓼、醋延胡索、三七加7倍量60%乙醇提取3次,每次1.5 h,过滤得醇提液;野菊花、夏枯草、茜草炭、粉萆薢加10倍量的水煎煮3次、每次1 h,滤过,滤液浓缩至相对密度为1.10~1.15(60℃)的浸膏,放冷,加乙醇使药液含醇量达70%,静置后过滤,得水提醇沉液;合并上述滤液,回收乙醇并浓缩至相对密度为1.20~1.25(60℃)。结论:该工艺简单、稳定可行,对花菊盆炎颗粒的工业化生产具有一定的参考价值。

菊花菊语

菊韵千古,淡雅的菊花在《诗经》中盛开,在屈原的《离骚》中盛开,在汉《乐府民歌》中盛开,在唐诗宋词元小曲、在泱泱中华五千年文明中粲然盛开。

Effect of Different Arbuscular Mycorrhizal Fungi Strains on Mineral Nutrition and Antioxidant Enzymes of Chrysanthemum morifolium

[Objective] The purpose was to study the effects of different arbuscular mycorrhizal fungi strains（AMF）on the mineral nutrition and antioxidant enzymes of Chrysanthemum morifolium.[Method] The pot experiment was conducted in the greenhouse,C.morifolium ＇Jinba＇ was inoculated with five kinds of AMF,N,P,K,malondial dehyde（MDA）content,as well as the superoxide dismutase（SOD）,peroxidase（POD） and catalase（CAT） activity in roots,leaves and petals of C.morifolium were measured at seedling and flowering stages.[Result] The G.i,G.e and G.m treatments could promote mineral nutrient absorption,increase N,P,K content in roots,leaves and petals of C.morifolium compared with the control without inoculation.The G.d、G.e and G.m treatments could significantly reduce MDA content in roots and petals,thus alleviating membrane permeability and lipid peroxidation.The G.i treatments could also improve the SOD,POD and CAT activities of C.morifolium,thereby increasing the capability of scavenging oxygen free radicals.[Conclusion] According to the comprehensive analysis,G.i was screened out as the best strain to improve mineral nutrition and antioxidant enzyme activities of C.morifolium.

Chemical constituents of the flower buds of Tussilago farfara

Aim To study the chemical constituents of the flower buds of Tussilago farfara L. in the China National GAP Base of Traditional Chinese Materia Medica and provide scientific basis for quality control. Methods The constituents were separated and purified by different chromatographic methods, and their structures were elucidated by IR, MS and NMR techniques. Results Twenty eight compounds were isolated from the flower buds of T. farfara. Their structures were identified as n- heptacosane （1）, bis（2-ethylhexyl）phthalate （2）, 7β-[3＇-ethylcrotonoyloxy]-1α-[2＇-methylbutyryloxy]-3,14-dehydro-Z-notonipetranone （3）, 7β-[3＇-ethylcrotonoyloxy]-1α-[2＇-methylbutyryloxy]-3,14-dehydro-E-notonipetranone （4）, tussilagone （5）, dibutyl phthalate （6）, bauer-7-ene-3β,16α-diol （7）, isobauerenol （8）, stigmasterol （9）, β-sitosterol （10）, 2,2-dimethyl-6-acetylchromanone （11）, n- hexadecanoic acid （12）, 7β-hydroxysitosterol （13）, 7α-hydroxysitosterol （14）, 7,14-bisdesacylnotonipetrone （15）, 2,3- dihydroxypropylpalmitate （16）, daucosterol （17）, 6-hydroxy-2,6-dimethylhept-2-en-4-one （18）, ferulic acid （19）, isoferulic acid （20）, caffeic acid （21）, α-D-glucose （22）, sucrose （23）, phthalic acid （24）, p-hydroxybenzoic acid （25）, gallic acid （26）, uridine （27）, and adenosine （28）. Conclusion Compounds 1, 12-16, 18 and 20 were obtained from the genus Tussilago for the first time.

Antioxidase Activities of Tetraploid Lines and Diploid Control Line of Chrysanthemum cinerariifolium

Objective The SOD, POD and APX enzyme activities of tetraploid lines and diploid control line of Chrysanthemum cinerariifolium in test-tube and field were determined in order to provide a reference for the selective breeding of high resistance good tetraploid varieties of C. cinerariifolium. Method The SOD, POD and APX activities of each sample were determined. Every material should have 3 replicates which been determined and calculated mean value to analyze. Result The various kinds of antioxidase activities in protective enzyme system of tetraploid lines were increased generally in comparison with that of diploid control line. And there is certain relativity between the samples in field and in test-tube of these lines. Conclusion The antioxidase activities of each line in test-tube could be considered as reference index to screen high resistance good tetraploid varieties of C. cinerariifolium.

菊花决明散加减防治角膜移植排斥反应作用观察

目的观察中药防治高危角膜移植免疫排斥反应的作用。方法总结了菊花决明散加减治疗角膜移植术后排斥反应24眼的临床情况，并进行了小样本的实验研究。结果临床患者用药7～14天结膜混合充血、角膜混浊和水肿得到不同程度改善；预实验结果证实中药组大鼠角膜移植后存活时间较对照组明显延长。结论复方菊花决明散具有缓解对角膜移植后免疫排斥反应的作用。

Effect of Sound Stimulation on the Lipid Physical States and Metabolism of Plasma Membrane from Chrysanthemum Callus

Plasma membrane vesicles of chrysanthemum ( Dendranthema morifolium (Ramat.) Tzvel.) callus was purified by sucrose gradient centrifugation to investigate the influence of sound stimulation on the lipid physical states and metabolism of plasma membrane. The results showed that sound stimulation decreased the content of phosphodiesters and the fluorescent intensity of DPH, but increased the light scattering value of the membrane, the fluorescent intensity of MC540 and the content of phosphomonoesters, indicating that the vesicles got looser, the charge density and hydrophobicity of membrane surface decreased under sound stimulation of some strength and frequency. However, the membrane fluidity increased under the condition. Meanwhile, the anabolism of membrane lipid increased and the catabolism decreased. It can be seen that the physical state and metabolism of membrane lipid is sensitive to sound stimulation.

Glycosides from Erigeron breviscapus

A new glycoside, gamma-pyranone-3-O-beta-D-[6'-(4'-hydroxy-3',5'-dimethyoxybenzoyl)]-glucopyranoside (erigeside D, 2), together with nine known compounds, viz., erigeside I (1), erigeside A (3), erigeside B (4), erigeside II (5), scopolin (6), icariside B-2 (7), blumenol C glucoside (8), (+)-syringaresinol O-beta-D-glucopyranoside (9) and scutellarein-7-O-beta-D-glucuronide methyl ester (10), was isolated from Erigeron breviscapus (Vant.) Hand.-Mazz. Their structures were established on the basis of spectral evidence. According to the beta-glucopyranosyl moiety in 7, the absolute configuration of 7 was determined by X-ray crystallographic analysis. Compounds 7, 8 and 9 were isolated for the first time from this genus, an compound 6 was first obtained from this plant.

GC-MS Analysis of Essential Oil Constituents from Buds of Tussilago farfara L.

Aim To analyse the constituents of the essential oils extracted from the buds of Tussilago farfara L. in the GAP Bases of Traditional Chinese Medical Materials and provide scientific basis for quality control. Methods The essential oils were extracted by water-steam distillation and separated by GC capillary column chromatography. The components were quantitatively determined by normalization, and identified by GC-MS. Results GC-MS exhibited 259 peaks and 65 compounds were identified, accounting for 84.62% of the total essential oil. Conclusion In the total essential oil contained in the buds of Tussilago farfara L., copaene （2.36%）, （ ＋ ） -Epi-bicyclosesquiphellandrene （ 3.91% ）, γ- elemene （2.18%）, fl-bisabolene （ 13.93% ）, spathulenol （ 3.44% ） as the sesquiterpenes and its derivatives, and 1-undecene （4.83%）, （ E）-cycloundecene （8.49%）, bicycle [ 10. 1.0] tridec-l-ene （ 1. 45% ）, 1-tridecene （3.44%）, （Z）-7,11-dimethyl-3-methylene-1,6,10-dodecatriene （2.66%）, 1- pentadecene （4.57%）, [ 1R-（ 1R^＊, 4Z, 9S^＊ ） ]-4,11,11-trimethyl-8-methylene-bicyclo [ 7.2.0] undec-4-ene （ 1.03% ）, 6,6-dimethyl-2-methylene-7-（ 3-oxobutylidene ）-oxepan-3-ylmethyl acetic acid ester （2.02%）, 1, E-11, Z-13-heptadecatriene （ 3.72% ）, （ Z, Z, Z） -9,12,15-octadecatrien-l-ol （ 1.85% ）, 3,7,11-trimethyl-dodeca-2,4,6,10-tetraenal （ 1.31% ）, n-hexadecanoic acid （ 3.12% ） , （Z, Z） -9,12-octadecadienoic acid （2.26%）, （ Z, Z, Z） -9,12,15-octadecatrienoic acid methyl ester （ 1.12% ） , heneicosane （ 1.82% ）, and pentacosane （ 1.03% ） are the main components.

Cloning and Analysis of the cDNA Sequence of XTH Gene from Chrysanthemum morifolium Petal

[Objective] This study aimed to investigate the structural character of xy- Ioglucan endotransglycosylase/hydrolase （XTH） gene in ethylene-insensitive feverfew. [Method] The total RNA was extracted from Chrysanthemum rnorifolium petal using Trizol reagent, and the cDNA fragment of XTH gene was cloned by RT-PCR and T/A cloning. [Result] The sequencing result showed that the cloned cDNA sequence was 911 bp. It was predicted to encode a polypeptide of 293 amino acids and had seven active sites of XTH family, and then named as CmXTH （gene accession number HM752243）. In addition, The BLAST analysis showed that the deduced amino acid sequence of CmXTH showed high homology with other 19 chosen plant XTHs. Among of these, CmXTH shared closer genetic relationship with Gerbera jamesonii, Solanum lycopersicum, whereas had relatively distant relationship with Populus euparatica, Fragaria ananassa, Actinidia deliciosa, etc. [Conclusion] The cloned fragment was certainly the cDNA of XTH gene, which was associated with the petal growth and senescence in Chrysanthemum morifolium.

Optimization on Identification Standards and Artificial Inoculation Methods In Vitro on Resistance to Chrysanthemum White Rust

[Objective] This research aimed at exploring an effective way for inoculation and identification of chrysanthemum white rust under controlled conditions. [Method] By combining the observation methods with the naked eye and under optical microscope, we had established the identification standards for chrysanthemum white rust with six classifications and optimized artificial inoculation methods in vitro. [Result] The results showed that bottled cuttings identification method and petri dished leaves identification method both can be used for identification in vitro of chrysanthemum white rust, bottled cuttings identification method had shown better effects than petri dished leaves identification method, and was supposed to be best artificial inoculation and identification method in vitro. [Conclusion] This research had provided a scientific method for safe and effective researches on chrysanthemum white rust, in order to control the occurrence and diffusion of this quarantine disease.

CHANGES OF ENDOGENOUS HORMONE CONTENTS DURING FLORAL BUD AND VEGETATIVE BUD DIF FERENTIATION IN THIN CELL LAYER CULTURE OF CICHORIUM INTYBUS L. EXPLANT

The sectioned thin cell layers (TCL) of flower stalk of Cichorium intybus L. were cultured in MS medium supplemented with NAA and BA or IAA and BA where floral and vegetative buds were developed from the explant. Endogenous IAA, DHZ+DHZR, iPA increased significantly during the floral bud formation, while Z+ZR remained changed. The levels of cytokinins, DHZ+DHZR, iPA, and Z+ZR all increased significantly during the vegetative bud formation, however IAA level was reduced during the first 7 days of culture and increased to two thirds of initial values on the day when the bud primordia were formed. The results suggested that the initiation of floral buds was associated with a high IAA/CTK ratio, whereas the induction of vegetative bud differentiation was related to a low IAA/CTK ratio.

Construction of Highly-efficient Direct Regeneration System of Different Chrysanthemum Species

Leaves and stems of 9 chrysanthemum species were taken as explants and inoculated to 6 differential media and root media to research effects of hor- mone combination and explants on chrysanthemum species. The results indicated that stem of 9 chrysanthemum species and leaf of 6 chrysanthemum species all showed higher regeneration capacity, with regeneration rate in 84%-100% and re- generation coefficient at 2.3-9.4. However, none adventitious buds were found differ- entiated from leaves of ＂28＂ and ＂5-17＂, and leaf regeneration capacity of ＂11＂ was lower, either. Most chrysanthemum species proved higher in stem regeneration ca- pacity, instead of leaf. Therefore, chrysanthemum species can be applied for rapid reproduction and genetic engineering.

江西出土的影青瓷选介

江西是产瓷名省,唐宋以来名窑迭出,尤以宋代景德镇窑的影青瓷开创了我国瓷坛的新篇章,产品质优工巧,釉色素雅,饮誉中外,这里择要介绍部分影青瓷珍品。 1、双系执壶 宋代。1989年南昌铁路公安分局公安处拨交。肩部有双系,可作提携。高12、口5.3、底径5.8厘米(封三,1)。 2、葫芦执壶 1985年乐安县宋嘉泰四年(1204)墓出土。釉色莹润,通体开片。壶把上与盖边有小圆钮可供系连。高15.6、口径2.2、底径5.4匣米(封三,2)。

Lamp Distribution Design for Adjustment of Florescence in Floral Production

[Objective] The aim was to introduce characters of light sources and select the optimal lamp scheme for floral production in greenhouses. [Method] With chrysanthemum cuttings as materials, HPSL, energy-saving fluorescent lamp, and LED agricultural lamp were made use of to carry out field tests in order to compare practical effects according to light characters of different lamps and plant growth de-mands. [Result] The results show that LED lamp performed the best of the three in practical use. LED square lamp designed in bat-wing shape would be the most ben-eficial for lamp distribution in a greenhouse. [Conclusion] LED agricultural lamp is the most popular currently, for it is energy saving and easy for operation.

Research on Pollen Grains of Tetraploid Lines and Diploid Control Line of Chrysanthemum cinerariifolium (Trev.) Vis.

[Objective] The aim was to study characters of pollen grains of tetraploid lines and diploid control line of Chrysanthemum cinerariifolium (Trev.) Vis.,morphological characters,fertility of pollen grain and germination percentage of seeds. [Method] Pollen grains were prepared by sulphuric acid-acetyl oxide decomposition method. The lengths of polar axis and equatorial axis of pollen grains were determined with general optical microscope. The morphology of pollen grains was observed with SEM (scanning electron microscope) and the typical visual fields of 2 500× (or 2 000×),7 000× were taken pictures. [Result] Comparing with the diploid control line,the pollen grains of five tetraploid lines which were tested were different from the diploid line in morphology,sculpture,etc.. 4 of the 5 tested samples were significant larger than the diploid line in size and one was similar to the diploid line. [Conclusion] This research provided references for breeding tetraploid improved varieties of Chrysanthemum cinerariifolium (Trev.) Vis. with good fertility and high germination percentage.

高台:“三变”带来“三美”

在湖北农垦草埠湖农场高台队,绿树掩映、鸟语花香,一排排、一幢幢龙脊凤尾、青瓦白墙、装饰崭新的房屋映入眼帘,这里绿树成荫、风景如画,置身其中,不禁让人想起“春游芳草地、夏赏绿荷池、秋饮桂花菊、冬吟白雪诗”的高远意境;它所散发的独特魅力,为我们勾勒出一幅幅恬静清馨的田园诗画。高台队地处长江支流沮漳河中游东岸,位于草埠湖农场镇头山北郊,全队面积7.7平方公里,耕地面积6763亩、人口1163人。

Induction of apoptosis and cell cycle arrest in human HCC MHCC97H cells with Chrysanthemum indicum extract

AIM： To investigate the effects of Chrysanthemum indicum extract （CIE） on inhibition of proliferation and on apoptosis, and the underlying mechanisms, in a human hepatocellular carcinoma （HCC） MHCC97H cell line. METHODS： Viable rat hepatocytes and human endothelial ECV304 cells were examined by trypan blue exclusion and MTT assay, respectively, as normal controls. The proliferation of MHCC97H cells was determined by MTT assay. The cellular morphology of MHCC97H cells was observed by phase contrast microscopy. Flow cytometry was performed to analyze cell apoptosis with annexin V/propidium iodide （PI）, mitochondrial membrane potential with rhodamine 123 and cell cycle with PI in MHCC97H cells. Apoptotic proteins such as cytochrome C, caspase-9, caspase-3 and cell cycle proteins, including P21 and CDK4, were measured by Western blotting. RESULTS： CIE inhibited proliferation of MHCC97H cells in a timeand dose-dependent manner without cytotoxicity in rat hepatocytes and human endothelial ceils. CIE induced apoptosis of MHCC97H cells in a concentration-dependent manner, as determined by flow cytometry. The apoptosis was accompanied by a decrease in mitochondrial membrane potential, release of cytochrome C and activation of caspase-9 and caspase-3. CIE arrested the cell cycle in the S phase by increasing P21 and decreasing CDK4 protein expression. CONCLUSION： CIE exerted a significant apoptotic effect through a mitochondrial pathway and arrested the cell cycle by regulation of cell cycle-related proteins in MHCC97H cells without an effect on normal cells. The cancer-specific selectivity shown in this study suggests that the plant extract could be a promising novel treatment for human cancer.

Inflammatory Mechanism of Total Flavonoids of Chrysanthemum and Medicated Serum on Castrated Dry Eye Animal and Cell Models

Objective To observe the effects of total flavonoids of chrysanthemum and medicated serum on the expression of related proteins in the lacrimal tissue and dry-eye cell models of male rabbits with dry eye caused by castration.Methods(1)150 male Japanese rabbits were randomly divided into five groups,with 30 rabbits in each group:normal control group(group A),sham group(group B),model group(group C),androgen control group(group D)and total flavonoids of chrysanthemum treatment group(group E).The androgen deficiency dry-eye model was established by bilateral castration in groups C,D and E.Normal saline was administered to groups A,B and C by gavage;androgen(testosterone propionate)was injected into muscle in group D;and group E was given total flavonoids of chrysanthemum by gavage.All white rabbits were tested the Schirmer I test(SIT)and tear break-up time(BUT).After euthanasia,tear gland tissue was harvested so that we could observe pathological changes in the expression of related inflammatory factors in the lacrimal gland tissue.The expression of interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α)and transforming growth factor-β1(TGF-β1)was detected in the lacrimal gland tissue by immunohistochemistry.Reverse transcription PCR was used to quantitatively detect expression of TGF-β1 mRNA.(2)Male Wistar rat lacrimal epithelial cells were used to establish a model of eye stem cell apoptosis caused by androgen levels.The blank control group was set up without androgen culture,the control group with androgen culture,and the total flavonoids of chrysanthemum group without androgen.The MTT method was used to determine the optimal intervention dosage of drug-containing plasma.Western blot and QPCR were used to detect the expression of AR mRNA,NF-κB phosphorylated protein and TGF-β1 in lacrimal epithelial cells,and the androgen-like effect of total flavonoids of chrysanthemum was observed.Results(1)Immunohistochemistry showed that groups A,B,D and E had significantly lower expression of IL-1βand TNF-αthan group C(P<0.05);among these,group E had slightly higher expression than group D(P>0.05).RT-PCR results showed that the relative expression of TGF-β1 mRNA in groups A,B,D and E was significantly higher than in group C(P<0.05),and the relative expression of TGF-β1 mRNA in groups D and E was higher than that in groups A and B(P<0.05).(2)Using the MTT method,the final concentration of interfering cells was calculated to be 13.2%.The expression of AR protein,NF-κB and TGF-β1 in the chrysanthemum flavonoid plasma intervention and testosterone propionate intervention groups was enhanced,and there were significant differences relative to the blank group(P<0.01).The expression level of NF-κB in the total flavonoid containing plasma intervention group was lower than that in the testosterone propionate intervention group(P<0.01).Conclusions The total flavonoids of chrysanthemum can inhibit IL-1βand TNF-αexpression in the lacrimal gland tissue of castrated male rabbits with dry eye to increase synthesis of TGF-β1 mRNA and TGF-β1,thereby inhibiting the inflammatory response.The medicated plasma with total flavonoids of chrysanthemum promotes expression of AR mRNA,upregulating expression of NF-κB,further promoting upregulation of TGF-β1 protein expression in lacrimal epithelial cells,inhibiting inflammation by regulating related proteins,and ultimately alleviating the symptoms of dry eye.