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??Chrysanthemum morifolium has a long history of culture and use in China. Due to different germplasm resources, producing areas, and processing methods, many cultivated varieties have formed now. The varieties and processing methods of C. morifolium are affected by economic interests and processing cost, which change gradually. On the basis of spot investigation and related literature study, the changes of the varieties and processing methods of C. morifolium were summarized in this paper. It will provide theoretical evidence for the culture, processing, quality evaluation, and clinical application of C. morifolium.     

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??OBJECTIVE To study the monthly dynamics of physical and chemical indexes in Achyranthis Bidentatae Radix(ABR, the dried root of Achyranthes bidentata Bl.) stored in simple and cool warehouses. METHODS ABR was stored in simple and cool warehouses for 27 months. The color was observed. The water content was determined based on the drying method. The contents of ??-ecdysone and 5-hydroxymethylfurfural (5-HMF) were determined by HPLC method. The accumulation of temperature difference between the simple and cool warehouses was evaluated with a relative temperature cumulation (RTC) method. The monthly dynamics of physical and chemical indexes of ABR was analyzed with RTC. RESULTS As the extension of storage time, the ABR stored in the simple warehouse showed deeper color and harder texture, but the ABR stored in the cool warehouse still had soft texture without significant color change. The contents of ??-ecdysone in ABR stored in the two warehouses both gradually decreased and dropped to lower than the limit of 0.030% ruled by China Pharmacopoeia when being stored for up to 27 months. The contents of 5-HMF of ABR stored in the two warehouses both increased and were higher for the sample in the simple warehouse than that in the cool warehouse. CONCLUSION The concept of RTC is put forward and used to study the monthly dynamics of chemical constituents in traditional Chinese medicine during storage for the first time. The physical and chemical indexes of ABR varies during storage. Two years of storage time of ABR is suggested.     

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??OBJECTIVE To study the coumarins from Mallotus conspurcatus. METHODS The ethyl acetate part of 75% ethanol extracts from Mallotus conspurcatus was isolated. The compounds were isolated and purified with column chromatography, and their structures were elucidated on the basis of ¹H-NMR, ¹³C-NMR and ESI-MS data. RESULTS Twelve coumarins were determined to be malloapelin A (1), 5??-demethyl aquillochin (2), aquillochin (3), cleomiscosin B (4), 6-hydroxy-7-methoxy-coumarin (5), braylin (6), aegelinol (7), bergapten (8), isofraxidin (9), fraxidin (10), grevillone (11), and fraxetin (12). CONCLUSION Compounds 6-12 are isolated from this genus for the first time.     

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??OBJECTIVE To study the effect and action mechanism of the large plant Rhodiola Tablet on myocardial ischemia of rats. METHODS Rat myocardial ischemia models were induced by coronary artery ligation. The levels of AST, ALT, CK and CK-MB in serum were observed, at the same time the expressions of B-raf, Ras and p-ERK1/2 in myocardial tissue were determined. RESULTS The levels of AST, ALT, CK and CK-MB in rat serum, as well as the B-raf, Ras and p-ERK1/2 expression in rats myocardium in the large plant Rhodiola group were significantly lower than that in the model group. CONCLUSION Large plant Rhodiola has a certain antagonism on myocardial ischemia injury in rats caused by coronary artery ligation. The mechanism of action may be related to the decreases in serum AST, ALT, CK and CK-MB,as well as myocardial high expression of the B-raf, Ras and p-ERK1/2.     

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??OBJECTIVE To provide the scientific basis for the application of mycorrhizal fungi in Dendrobium officinale artificial cultivation. METHODS The dominant strain was screened in a planting base by using biomass as the index. The effects of initial inoculums and supplementary inoculation on the growth and the chemical composition of D. officinale were further investigated. RESULTS Mycorrhizal fungi M2(Mycena sp.) was found to be the advantageous strain. After 16 months of growth, the stem dry weight(SDW) and plant dry weight(PDW) of the three M2-treatment groups were significantly higher than those of CK, and the SDW of group M(M2 initial inoculums of 1.0 g??clump^-1) was the highest, which increased by 39.2% and 11.1% compared with CK and group L(M2 initial inoculums of 1.0 g??clump^-1)(P<0.05), respectively. Furthermore, M2 had no effect on the chemical properties of stem polysaccharide, and could improve the contents of naringenin and bibenzyls in the stems. The supplementary inoculation of M2 enhanced the plant-growth-promoting effects of M2. The SDW of the supplementary group increased by 18.0%(P<0.05) compared with the non-supplementary group, while supplementary inoculation had a certain effect on the chemical properties of polysaccharide. CONCLUSION Mycorrhizal fungus M2 has good application prospects for D. officinale artificial cultivation.     

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??OBJECTIVE To establish an HPLC-UV-ESI-MSⁿ method for the study of impurity profile of amoxicillin and clavulanate potassium tablets. METHODS Agilent 1100 LC/MSD Trap liquid chromatography-mass spectrometry was used, and the column was Shim pack CLC-ODS RP18(4.6 mm??250 mm, 5 ??m). The mobile phase A was 20 mmol??L^-1 ammonium acetate (pH adjusted to 6.0), and the mobile phase B was 20 mmol??L^-1 ammonium acetate-acetonitrile (20??80) (pH adjusted to 6.0). Gradient elution was performed at a flow rate of 1.0 mL??min^-1. ESI source was used. Positive and negative ion scan was conducted with a scanning range of m/z 100-1 500. The nebulizing pressure was 275.8 kPa, dry gas flow was 9 L??min^-1, and post-column diversion ratio was 1??5. Some related substances were identified by comparing the retention time in the chromatography, [M+H]⁺ spectrum and MS² spectrum with those of the reference substances, while the others which do not have reference substances were deduced or speculated by analyzing the MS² or MSⁿ fragmentation with the help of a rule summarized from the MS² fragmentation of amoxicillin, clavulanic acid and system suitability impurity reference substances. RESULTS A total of 15 related substances were separated and characterized including nine known impurities like amoxicilloic acid, amoxicillin dimer, etc. and six unknown impurities. CONCLUSION The method can be applied in the identification and qualitative analysis of the related substances in amoxicillin and clavulanate potassium tablets and is helpful for the quality control and optimization of the synthetic process.     

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??OBJECTIVE To study the effects of 5-Fu combined with resveratrol (Res) on the growth and apoptosis of A431 and TE-1 cell lines and the underlying mechanism.To study the therapeutic effects of 5-Fu/Res combination on mouse skin papilloma chemically induced by DMBA/TPA. METHODS The effects of 5-Fu/Res combination on the viability of cancer cells were evaluated by MTT assay, growth curves assay and LDH releasing assay. The inhibitory effect of combination of the two drugs was analyzed by the method of Chou and Talalay. Apoptoses of A431 and TE-1 were determined by inverted microscope and gel electrophoresis of DNA fragment analysis. Intracellular Ca²⁺ concentration was determined to study the underlying mechanisms of 5-Fu and Res on the apoptosis of cancer cells. The mouse skin papilloma model was established by DMBA and TPA,the expression of actived-caspase-3 in mouse skin was examined by immunohistochemistry. RESULTS Combination of 5-Fu and Res decreased the median inhibitory concentration (IC₅₀) to cancer cells remarkably. 5-Fu/Res combination showed a synergistic effect on apoptosis of A431 and TE-1 cells. Much more typical morphological changes of apoptosis and amount of fragmented DNA were observed in the cells treated with 5-Fu and resveratrol in combination than that in the cells treated with the agents alone. The increase of [Ca²⁺]_i induced by 5-Fu/Res combination might be involved in the apoptotic induction. There was a significant decrease of the number of tumors after treatment with 5-Fu and Res in the tumor-bearing mice model. Active caspase-3 in the cells of mice skin was generated by 5-Fu in combination resveratrol was more effective than either alone. CONCLUSION The 5-Fu/Res combination shows synergistic anti-tumor effects both in vitro and in vivo.     

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??OBJECTIVE To investigate the liposoluble constituents from the herb of Pothos chinensis(Raf.)Merr. of Araceae. METHODS The compounds were isolated and purified by silica gel, Sephadex LH-20, semi-preparative HPLC and recrystallization methods. Their structures were identified on the basis of spectral analysis, physicochemical characteristics and comparison of their spectral data with those in the literature. RESULTS Seventeen compounds were isolated and identified respectively as honokiol(1), hinokinin(2), racemosol(3), 24-propylcholest-7-en-3-ol(4), ??-sitosterol(5), daucosterol(6), hentriacontane(7), palmitic acid(8), oleic acid(9), linoleic acid(10), 9, 12, 15-ocatadecatrienoic acid(11), 13-phenyltridecanoic acid(12), 13-(3??, 4??-methylenedioxyphenyl)-tridecanoic acid(13),1- monostearin(14), 1-monopalmitin(15), benzoic acid(16), and vanillic acid(17). CONCLUSION Compounds 1-16 are isolated for the first time from this plant.
     

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??OBJECTIVE To investigate the antioxidant activities and chemical constituents of Polygonum chinese L. var hispidum Hook.f.METHODS The compounds were isolated by chromatographic techniques with silica gel, Sephadex LH-20 and semi-preparative HPLC. Their structures were identified by analysis of physical and spectral data, and confirmed by comparison of their spectral data with the reported values in the literature or those of authentic samples. The antioxidant activities of the extractions of Polygonum chinese L. var hispidum Hook.f. were evaluated by oxygen radical absorption capacity(ORAC) assay. RESULTS Fifteen compounds were isolated and identified as isopsoralen(1), 2,3,4,6,7-pentamethoxy-9,10-dihydrophenanthrene(2), 1,5,7-tri hydroxy-3-methyl-anthraquinone(3), quercetin(4), gallincin(5), gallic acid(6), heneicosanoic acid(7), nonadecanoic acid(8), arachidic acid(9), triacontanoic acid(10), tetracosanoic acid(11), palmitic acid(12), lacceroic acid(13), ??-sitosterol(14), and daucosterol(15), respectively. Ethyl acetate extracts had stronger antioxidant activities. CONCLUSION All compounds are isolated from the species for the first time, and compound 2 is obtained from this genus for the first time     

铁皮石斛化学成分研究

目的研究铁皮石斛Dendrobium officinale的化学成分。方法使用硅胶色谱法、反相硅胶色谱法及半制备HPLC等技术进行分离纯化,采用现代波谱技术鉴定化合物结构。结果从铁皮石斛70%乙醇提取物中分离得到14个化合物,分别鉴定为苯甲醇-O-β-D-吡喃葡萄糖苷(1)、异鼠李素-3-O-α-L-鼠李糖基(1→2)-β-D-吡喃葡萄糖苷(2)、conicaoside(3)、3,4,5-三甲氧基苯-1-O-β-D-吡喃葡萄糖苷(4)、2,6-二甲氧基-4-羟基苯酚-1-O-β-葡萄糖苷(5)、4-(3'-羟丙基)-2,6-二甲氧基苯酚-3'-O-β-D-葡萄糖苷(6)、胸腺嘧啶脱氧核苷(7)、紫丁香苷(8)、4-烯丙基-2,6-二甲氧基苯基葡萄糖苷(9)、(+)-丁香脂素(10)、丁香脂素-4'-O-β-D-单葡糖苷(11)、柚皮素(12)、对羟基苯甲酸(13)、4,4'-二羟基-3,5-二甲氧基联苄(14)。结论化合物1~6为首次从石斛属植物中发现,7~8为首次从本植物中发现。     

铁皮石斛与霍山石斛中甘露糖、葡萄糖及柚皮素的含量比较

目的:优化铁皮石斛中甘露糖与葡萄糖的柱前衍生HPLC含量测定方法以及柚皮素HPLC含量测定方法,比较铁皮石斛与霍山石斛中这3种成分的含量差异。方法:在2015年版《中国药典》铁皮石斛甘露糖柱前衍生HPLC含量测定项下色谱条件基础上,选择乙腈-0. 02 mol·L~(-1)乙酸铵溶液为流动相系统梯度洗脱,同时测定甘露糖与葡萄糖的含量,并分析甘露糖与葡萄糖的峰面积比值;采用Kromasil 100-5 C18色谱柱(4. 6 mm×250 mm,5μm);检测波长250 nm;流速1. 0 mL·min~(-1);柱温30℃。柚皮素HPLC含量测定采用Kromasil 100-5 C18色谱柱(4. 6 mm×250 mm,5μm);流动相乙腈-甲醇-0. 4%磷酸溶液,梯度洗脱;检测波长290 nm;流速0. 8 mL·min~(-1);柱温40℃。结果:甘露糖与葡萄糖在0. 15~3. 0,0. 075~2. 25μg线性关系良好(r=0. 999 9),平均加样回收率分别为99. 01%(RSD 2. 1%),101. 69%(RSD 2. 0%),重复性、耐用性等其他方法学研究符合要求。43批不同产区铁皮石斛中甘露糖、葡萄糖以及两者的含量之和分别在12. 75%~36. 40%,2. 93%~18. 39%,19. 23%~54. 58%,除极少数样品外,基本符合2015年版《中国药典》甘露糖含量限度要求,甘露糖与葡萄糖含量之和也接近总多糖含量限度要求;含量与产区相关性不显著。12批霍山石斛的总糖则分别在14. 33%~29. 47%,6. 64%~15. 20%,25. 73%~44. 37%,其含量以及峰面积比值基本落在铁皮石斛范围期间,多批次的平均含量也基本与铁皮石斛一致(约33%左右)。柚皮素在0. 020 8~0. 832 0μg线性关系良好(r=0. 999 9),平均加样回收率为101. 96%(RSD 1. 8%)。11批铁皮石斛与7批霍山石斛的柚皮素含量分别为0. 053 2~0. 122 4 mg·g~(-1)(均值为0. 081 0 mg·g~(-1)),0. 040 3~0. 090 0 mg·g~(-1)(均值为0. 068 3 mg·g~(-1)),铁皮石斛含量稍高于霍山石斛,但含量均亦未达到0. 02%的质量标准下限的常规要求。结论:铁皮石斛中甘露糖与葡萄糖HPLC含量测定方法重复性较好,用两者含量之和替代具有较大误差的总多糖含量作为测定指标具有可行性;单糖含量测定可应用于霍山石斛的定量质控指标;但依据2种石斛的总多糖含量、水解后的单糖含量与峰面积比值以及柚皮素含量,无法区分铁皮石斛与霍山石斛,需结合其他专属性方法方能对两种石斛进行区别。     

合江金钗石斛不同品系农艺性状比较与品质分析

目的:对合江金钗石斛3个品系(JC-1,JC-2,JC-3)主要农艺性状与品质进行比较,为合江金钗石斛生产提供指导,为金钗石斛新品种选育提供参考依据。方法:用直尺、游标卡尺和分析天平等测定了合江金钗石斛的主要农艺性状与产量性状;采用苯酚-硫酸比色法和酸性染料比色法分别测定茎中多糖与总生物碱含量。结果:2年生的JC-1在茎节数、节间长、茎鲜重与茎干重方面表现最佳。从茎长来看,无论是1年生,2年生还是3年生,均表现为JC-1JC-2JC-3。茎粗则随着年限的增长而增粗,JC-2粗于JC-1和JC-3,且JC-2从2年生到3年生这个阶段增长迅速。折干率均表现为2年生大于1年生与3年生,JC-1的折干率远大于JC-2与JC-3。JC-3株型最小,折干率最低。在有效成分含量方面,多糖质量分数为JC-1(4.64%)JC-2(3.02%)JC-3(2.20%);总生物碱质量分数为JC-3(0.59%)JC-2(0.48%)JC-1(0.45%),但折算为茎鲜品中多糖与总生物碱含量均为JC-1最高。结论:综合生产及品质等角度,笔者认为合江金钗石斛品系JC-1最适合推广种植。     

铁皮石斛叶的化学成分研究

目的研究兰科植物铁皮石斛Dendrobiumofficinale叶的化学成分。方法运用Sephadex LH-20、MCI CHP-20P、ODS等柱色谱和高效液相色谱技术进行分离纯化,并根据理化性质和NMR、MS波谱数据鉴定化合物结构。结果从铁皮石斛叶中分离得到24个化合物,分别鉴定为3,4-二羟基-5,4′-二甲氧基联苄(1)、杓唇石斛素(2)、4,4′-二羟基-3,5-二甲氧基联苄(3)、densiflorol A(4)、(S)-3,4,α-三羟基-5,4′-二甲氧基联苄(5)、石斛酚(6)、dendrocandin U(7)、dendrocandin B(8)、黑麦草内酯(9)、(6R,9S)-6,9-二羟基-4,7-巨豆二烯-3-酮-9-氧-β-D-吡喃葡萄糖苷(10)、(6R,9S)-9-羟基-4,7-巨豆二烯-3-酮-9-氧-β-D-吡喃葡萄糖苷(11)、(+)-丁香脂素(12)、芦丁(13)、2-羟基苯并噻唑(14)、对羟基苯乙酮(15)、对羟基苯甲酸(16)、原儿茶酸(17)、儿茶酚(18)、对羟基苯丙酸乙酯(19)、甘油亚麻酸酯(20)、亚麻酸2-丁氧基乙酯(21)、棕榈酸(22)、十八碳癸二烯酸-2,3-二羟基丙酯(23)和urticifolene(24)。结论化合物10、11、14～23为首次从兰科植物中分离得到,化合物1、2、4和6为首次从该植物中分离得到。     

铁皮石斛原球茎高效再生体系的研究

目的以铁皮石斛Dendrobium officinale实生种子为外植体,研究铁皮石斛原球茎的增殖、分化、诱导生根等条件,筛选出适宜铁皮石斛原球茎高效再生的诱导条件,建立高效的再生体系。方法以铁皮石斛种子为外植体,在培养基上诱导产生原球茎,经过增殖培养后,转移到分化培养基中诱导芽苗分化,待分化的芽苗长到1~2 cm时,转移到生根培养基诱导生根,最终长成完整的再生植株。结果适宜铁皮石斛生长的基本培养基为1/2 MS;原球茎诱导的培养基为1/2 MS+1.0 mg/L6-BA+0.2 mg/L NAA+50 g/L土豆泥;原球茎增殖的最适培养基为1/2 MS+1.0 mg/L 6-BA+0.5 mg/L NAA,最高增殖系数达23;最佳分化培养基为1/2 MS+2.0 mg/L 6-BA+0.2 mg/L NAA,平均分化率最高达95%;最佳生根培养基为1/2 MS+0.3mg/L NAA+50 g/L土豆泥,生根率达100%。结论铁皮石斛实生种子是铁皮石斛离体繁殖的优良外植体来源。以铁皮石斛种子诱导获得的高效再生技术体系可以为铁皮石斛快速繁殖和工厂化生产提供理论基础。     

铁皮石斛快繁技术体系研究

目的以铁皮石斛Dendrobium officinale带芽茎段为材料,对铁皮石斛的组织培养进行系统研究,以期建立铁皮石斛的快繁技术体系。方法采用植物组织培养的方法对铁皮石斛外植体的消毒方法、原球茎的诱导、增殖、分化、幼苗生根及瓶苗移栽等进行研究。结果铁皮石斛外植体最佳的消毒方式为75%乙醇消毒30 s,再用0.1%HgCl2消毒10 min;铁皮石斛带芽茎段原球茎诱导的最佳培养基为MS+1.0 mg/L 6-BA+0.5 mg/L NAA+2 g/LAC,诱导率达到34.98%;原球茎增殖的最佳培养基为MS+0.5 mg/L 6-BA+0.8 mg/L 2,4-D+2 g/LAC,增殖率达到89.6%;原球茎分化的最佳培养基为MS+1.2mg/L 6-BA+0.2 mg/L IBA+2 g/L AC,分化率达到89.6%;幼苗生根的最佳培养基配方为1/2 MS+2.5 mg/L NAA+15%土豆汁+2 g/LAC,生根率达到90.4%～92.8%;瓶苗移栽最佳方式为大苗、树皮块苔藓草做基质、基质中添加0.03 mg/L赤霉素,并接种适量菌根真菌Epulorhiza sp.。结论建立了铁皮石斛的快繁技术体系,为铁皮石斛的工业化生产奠定技术基础。     

铁皮石斛化学成分、药理作用及其质量标志物(Q-marker)的预测分析

铁皮石斛是中国传统名贵中药,属补益药中的补阴药,具益胃生津、滋阴清热等功效,其质量标准的建立一直是热点研究问题。通过化学成分和药理活性这2个方面对铁皮石斛研究现状进行综述,并在此基础上基于中药质量标志物(Q-marker)的概念,分析其生源途径、传统功效、现代药理作用与化学成分之间的关系,对铁皮石斛的Q-marker进行预测分析,为建立和完善其质量标准提供依据。     

铁皮石斛及金钗石斛菌根真菌的分离及其生物活性测定

目的 :对我国云南、四川等地采集的野生铁皮石斛Dendrobiumcandidum和金钗石斛D .nobile根中的内生真菌进行分离并测定其生物活性。方法 :采用平板分离法分离内生真菌 ;所获得真菌分别与铁皮石斛种子、铁皮石斛及金钗石斛无菌苗共培养测定其生物活性。结果 :分离获得内生真菌 2 5种 ,主要属于担子菌Basidiomy cotina和半知菌Deuteromycotina;生物活性测定结果表明 ,其中 5种真菌可促进铁皮石斛种子萌发 ;7种真菌可与铁皮石斛和金钗石斛幼苗形成共生关系 ,但仅有 3种真菌对幼菌有促生作用。结论 :分离和筛选促进石斛生长发育的适宜菌根真菌 ,是菌根技术应用于石斛生产的关键问题之一。     

铁皮石斛内生真菌Verticillium sp. KY-18的化学成分研究

目的研究铁皮石斛Dendrobium officinale内生真菌Verticillium sp.KY-18中氧杂环类化合物。方法采用色谱分离技术进行分离和纯化,并根据谱学数据鉴定化合物的结构。结果从Verticillium sp.KY-18的发酵产物中分离得到13个氧杂环类化合物,分别鉴定为2,6-dihydroxy-2-methyl-7-(prop-1E-enyl)-1-benzofuran-3(2H)-one(1)、nigrosporapyrone D(2)、oosponol(3)、2-甲基-4-吡喃酮(4)、6-甲基-5-[(1E)-丁烯基]-2-吡喃酮(5)、penicisochroman D(6)、verrucosapyrone B(7)、(1S,3S)-1,8-dimethoxy-3,5-dimethyl-6-hydroxyisochroman(8)、phomopsinone A(9)、pseudohalonectrin A(10)、dictafolin-A(11)、2,3-dihydro-5,7-dihydroxy-2,6,8-trimethyl-4H-1-benzopyran-4-one(12)、2-pyrone-4,6-dicarboxylic acid(13)。结论化合物2~10为首次从该属真菌中分离得到;化合物11~13为首次从该真菌中分离得到。