完达山地区槲寄生(Viscum coloratum)种群特征研究

于2012年4-11月对老平岗林场6个群落中7种寄主上的槲寄生(Viscum coloratum)种群进行了研究,结果表明:以寄主为单位,槲寄生的最终分布格局取决于寄主的格局;而在每个寄主上则呈聚集性分布。从数量上看,寄生强度为:山杨"白桦"辽东桤木"黑桦"紫椴=青楷槭"色木槭;各寄主上单株槲寄生平均结果量为:山杨"白桦"辽东桤木"青楷槭"紫椴"色木槭"黑桦;红黄两种果实型植株的比例近于1∶1。从遗传上看,槲寄生表现出过量繁殖特征;其种子传播与太平鸟存在密切联系,经鸟类粪便排出的种子82%落在地面,15%黏附在下层灌木上,3%附着在寄主树干上。本研究认为:寄主的分布格局,寄主所在群落的组成及发育,传播者的生活习性和槲寄生自身的繁殖策略是影响其种群的主要因素。

Development of the fingerprints for the quality evaluation of Viscum coloratum by high performance liquid chromatography

A high-performance liquid chromatography coupled ultraviolet （HPLC-UV） method was developed for a chemical fingerprint analysis of Viscum coloratura. Eighteen peaks were selected as the common peaks and Homoeriodictyol-7-O-β-D-apiosiyl-（1→2）-β-D-glucoside was used as a reference. The relative areas of common peaks were used for hierarchical clustering analysis and similarity calculation. Thirty-seven samples collected from different sources were classified into five groups. The similarities of 21 batches Viscum coloratura samples were beyond 0.90. The results obtained suggest that the chromatographic fingerprint can efficiently identify Viscum coloratum. Additionally, the fingerprints can then be used to evaluate the correlation between Viscum coloratura and hosts.

Purification and Primary Structure Determination of a Novel Polypeptide Isolated from Mistletoe Viscum coloratum

A novel polypeptide was isolated from mistletoe Viscum coloratum. The primary structure of the polypeptide named viscotoxin B2 was determined to be KSCCKNTTGRNIYNT CRFAGGSRERCAKLSGCKIISASTCPSDYPK by Edman degradation. Viscotoxin B2 shared high sequence homology with viscotoxins isolated from Viscum album. Pharmacological experiments showed that viscotoxin B2 had distinct cytotoxic activity on tumor cells. Viscotoxin B2 could be used as a leading compound in cancer therapy.

Metabolomics-based comparative analysis of the effects of host and environment on Viscum coloratum metabolites and antioxidative activities

Viscum coloratum(Kom.)Nakai is a well-known medicinal hemiparasite widely distributed in Asia.The synthesis and accumulation of its metabolites are affected by both environmental factors and the host plants,while the latter of which is usually overlooked.The purpose of this study was to comprehensively evaluate the effects of host and habitat on the metabolites in V.coloratum through multiple chemical and biological approaches.The metabolite profile of V.coloratum harvested from three different host plants in two habitats were determined by multiple chemical methods including high-performance liquid chromatography-ultraviolet(HPLC-UV),gas chromatography-flame ionization detector(GC-FID)and ultra-performance liquid chromatography quadrupole time of flight mass spectrometry(UPLC-QTOF/MS).The differences in antioxidant efficacy of V.coloratum were determined based on multiple in vitro models.The multivariate statistical analysis and data fusion strategy were applied to analyze the differences in metabolite profile and antioxidant activity of V.coloratum.Results indicated that the metabolite profile obtained by various chemical approaches was simultaneously affected by host and environment factors,and the environment plays a key role.Meanwhile,three main differential metabolites between two environment groups were identified.The results of antioxidant assay indicated that the environment has greater effects on the biological activity of V.coloratum than the host.Therefore,we conclude that the integration of various chemical and biological approaches combined with multivariate statistical and data fusion analysis,which can determine the influences of host plant and habitat on the metabolites,is a powerful strategy to control the quality of semi-parasitic herbal medicine.

First report of leaf-spot disease caused by Sphaeropsis visci on Asian mistletoe [Viscum coloratum (Kom.) Nakai] in China

In January 2017, symptoms of a leaf-spot disease were observed on Viscum coloratum plants in Yichun,China. The infected leaves were chlorotic, while sunken lesions formed on diseased branches, which were initially light brown and later turned dark brown. Moreover, diseased branches and leaves formed semi-buried, small, and spherical pycnidia. In total, 20 leaves and 19 branches diseased samples from Yichun, China were collected and examined. The fungus was isolated and identified using Koch’s postulates, morphological characteristics and DNA sequence data. The data presented herein confirmed that the pathogen responsible for the disease was Sphaeropsis visci. To the best of our knowledge, this is the first report of it in China.

A novel plant defensin from Chinese mistletoe,Viscum coloratum (Kom.) Nakai

A novel polypeptide named as defensin CM was isolated from Chinese mistletoe, Viscum coloratura （Kom.） Nakai. The amino acid sequence was determined by the combination of Edman degradation, endoproteinase Lys-C digestion, and MALDI-TOF mass spectrometry. The primary structure was ATCSAPSGRF KGACFSSNTC SNICKTLEGL KDGHCTGLAC YCSRNC.

Long-term survival of a patient with advanced pancreatic cancer under adjunct treatment with Viscum album extracts: A case report

BACKGROUND Advanced pancreatic cancer(aPC) has a poor prognosis with limited survival benefit from current standard treatment. Viscum album extracts(VAE) are used by many cancer patients, showing immune-stimulating effects, improved quality of life, and a survival benefit in patients with aPC.CASE SUMMARY A 59-year-old architect developed epigastric pain. A cystic lesion of the pancreas of 45-mm diameter was detected. In a follow-up magnetic resonance imaging,about one year later, multiple lesions were seen in the corpus and the tail of the pancreas; CA-19-9 was elevated to 58.5 U/mL. A distal pancreatectomy with splenectomy was performed, and a tumor of 7 cm × 5 cm × 3.5 cm was excised.Histologic investigation showed an intraductal papillary mucinous neoplasmassociated invasive adenocarcinoma with invasion of the lymph vessels,perineural invasion, and positive nodes(2/27); surgical margins showed tumor cells, and the tumor was classified as pT3 N1 M0 R1. The patient was treated with radiation of the tumor bed and capecitabine/oxaliplatin followed by gemcitabine and FOLFIRINOX. Seven months after surgery, a liver metastasis was detected and treatment with FOLFIRINOX was started. Four months after detection of the metastasis, the patient opted for additional treatment with VAE. Another month later, the metastasis was treated with radiofrequency ablation(RFA). Eight months later, the hepatic lesion recurred and was again treated with RFA. Thecontinuous VAE treatment was increased in dose, and the patient stayed recurrence-free for the next 39 mo in good health and working full-time(as of the time this case report was written).CONCLUSION We present the case of a patient with aPC with R1-resection with development of liver metastasis during the course of treatment who showed an overall survival of 63 mo and a relapse-free survival of 39 mo under increasing VAE therapy. The possible synergistic effect on tumor control of RFA treatment and immunestimulatory effects of VAE should be further investigated.

Study on chemical composition of Viscum coloratum and its HPLC fingerprint in different harvest periods

The purpose of this study was to evaluate the similarity of chemical profile of Viscum loloratum harvested in different months using high performance liquid chromatography(HPLC)fingerprint and multivariate statistical analysis,and to determine the optimal harvest time of Viscum coloratum.8 compounds were isolated from Viscum coloratum by a variety of isolation techniques,3 of which were isolated from the genus Viscum L.for the first time.The fingerprint of Viscum coloratum was established by HPLC,and 16 common peaks were obtained,4 of which were identified by comparison with the reference substances.Results showed that the Viscum coloratum harvested in 6 months could be roughly divided into three groups based on the chemical profiles,and the Viscum coloratum harvested in January had higher levels of the marked compounds.The HPLC fingerprint objectively reveals the characteristics of Viscum coloratum harvested in different months,which can be used to evaluate and control the quality of Viscum coloratum harvested in different months.

Proximate and mineral components of Viscum album var. coloratum grown on eight different host tree species

Mistletoes are parasite plants growing on various tree species,taking nutrients from the host.Beneficial compounds of mistletoe were used in medicine and these mistletoes were commonly collected from oak species that were not well identified.We analyzed the nutritional value by measuring proximate and mineral components of Korean mistletoe(Viscum album var.coloratum)from various host trees.Carbohydrate was the most proximate component in both leaves and twigs followed crude protein,crude fiber,crude ash and crude fat.While the contents of the proximate components and minerals varied among hosts and between plant parts,leaves contained higher crude protein and ash,and more minerals except for magnesium than twigs,although no significant difference was found inmistletoe grown on Prunus mandshurica.There were significant differences of proximate and mineral contents depending on the host species.Mistletoe grown on Chaenomeles sinensis contained higher levels of carbohydrates,crude fat,crude ash,magnesium,calcium,sodium,iron,and zinc than mistletoe grown on other species.Mistletoe grown on P.mandshurica contained high mineral levels of potassium,copper and phosphorous.The nutritional value of mistletoes were greatly influenced by the host species but among the hosts,specific nutrients accumulated in leaves more than in twigs,and vice versa.Based on the analysis,Korean mistletoe contains relatively good quality of nutrients and has high potential for dietary supplements as well as for medicinal compounds.

New Polypeptides from Chinese Mistletoe, Viscum coloratum (Kom.) Nakai

Three viscotoxins have been isolated from Chinese mistletoe, Viscum coloratura （Kom.） Nakai. The primary structures were determined unambiguously by the combination of Edman degradation, endoproteinase Elu-c digestion, and Q-TOF mass spectrometry. One was identified as viscotoxin C1, which was for the first time isolated from Chinese mistletoe. The other two were new polypeptides, named as viscotoxin B5 and viscotoxin B8. Pharmacological studies showed that the polypeptides exhibit distinct cytotoxicity to human cancer cells.

Efficacy and safety of viscum fraxini-2 in advanced hepatocellular carcinoma:a phase Ⅱ study

Objective： New systemic therapies are needed to improve the prognosis of patients with advanced-stage hepatocellular carcinoma. The study was conducted to determine the efficacy and safety of viscum fraxini-2 in advanced Hepatocellular carcinoma. Methods： A phase II study with a two-stage design that enrolled a total of 120 patients with chemotherapy nai＇ve advanced hepatocellular carcinoma. The mistletoe preparation for the study is an aqueous injectable solution that contains one milliliter of viscum fraxini. Two ampoules of viscum fraxini were administered subcutaneously once weekly. Re- suits： Chronic hepatitis C virus infection was the predominant cause of liver disease （60%） in studied cases. According to the response evaluation criteria in solid tumors 24 patients （20%） achieved objective response （including 2 complete responses） and 40 patients （33.3%） achieved stable disease. The median progression free survival for all patients was 4 months （range 1-28 months; 95% CI 3.3：4.7 months）. The median overall survival for all patients was 8 months （range 1-28 months; 95% CI 6.3：9.7 months）. The median survival for patients who achieved responsive or stable disease was 16 months. The toxicity was generally mild and well tolerated, mainly in the form of local reaction and fever. There were no drug related discontinu- ation or toxic deaths. Conclusion： Viscum fraxini-2 is an effective, safe treatment for patients with advanced hepatocellular carcinoma. Further randomized controlled trials are recommended.

Renoprotective and Blood Pressure Lowering Impact of Equisetum arvense and Viscum album Therapy in Experimental Model of Chronic Kidney Disease

Background: Chronic kidney disease (CKD) is an irreversible decline in the glomerular filtration due to nephrosclerosis and glomerular loss. Rat remnant kidney model enables to assess the benefits of different treatment possibilities. Aim: The aim of our study was to investigate renal morphology and functioning after 12 weeks of treatment with Equisetum arvense and Viscum album. Methods: Male Wistar rats with 5/6 nephrectomy received herbal drug preparation Equisetum/Viscum in a dosage of 0.007 g/kg/die (herbal group) or losartan (180 mg/l, ARB group) or remained untreated. Systolic blood pressure (SBP) and proteinuria were measured. Renal cortex tissue samples examined for focal-segmental glomerulosclerosis (FSGS) and interstitial fibrosis (IF). mRNA was isolated to estimate CCL2/MCP-1 gene transcription. Results: SBP was significantly lower both in herbal group and ARB group compared with untreated animals (p between ARB group and untreated NPX (p = 0.001). Quantitative RT-PCR analysis revealed statistically significant differences of mRNA transcription for MCP-1 between herbal group and untreated group (p Conclusions: We report about beneficial impact of horsetail Equisetum arvense and mistletoe Viscum album treatment on kidney functioning and morphology.

Mild Steel Corrosion Mitigation in Sulphuric Acid via Benign Isolated Phytochemicals from <I>Viscum album</I>

We present the evaluation of four Eco-friendly corrosion inhibitors for the corrosion mitigation of mild steel in acid media. The corrosion inhibition of mild steel by ethanol extracts from Viscum album (EEVA) and some of its isolated phytochemical components;phytates extract (PEVA), saponins extract (SEVA), and flavonoids extracts (FEVA) have been studied using gravi-metric and gasometric techniques. The results of the study reveal that these ecofriendly and benign extracts function as good inhibitors for mild steel corrosion in sulphuric acid. Inhibition efficiency of the extracts increases with inhibitor concentration and temperature rise. The trend of inhibition efficiency in lower inhibitor concentration is EEVA > PEVA > SEVA > FEVA and at higher concentration, the order was SEVA > PEVA > EEVA > FEVA. The presence of the plant extracts decreases the corrosion activation energy in the solution which indicates chemical adsorption mechanism. The adsorption of the components of the extracts is consistent with Temkin isotherm. The interaction between the isolated extracts is synergistic at lower inhibitor concentration and antagonistic at higher concentrations. The Kads values for PEVA and SEVA are higher than those of EEVA and FEVA. This implies that PEVA and SEVA are more efficiently adsorbed on the mild steel surface.

Study on Protection Mechanism of Viscum coloratum (Kom.) Nakai f. lutescens kitag Fruit Polysaccharides on Mice with Acute Alcoholic Liver Injury

This study was conducted to investigate the protective mechanism of V. coloratum fruit polysaccharides（ VCFP） on mice with acute alcoholic liver injury. Acute liver injury model was built by one-time alcohol gavage. The mice were randomly divided into VCFP-treated groups（ low-,medium-,and highdose）,alcohol model group and normal control group at the same time. VCFP-treated groups were administrated polysaccharide intragastrically continuously for4 weeks; and the alcohol model group and normal control group were administrated intragastrically the same amount of normal saline. The general situation and liver tissue morphological structure changes were observed. The results showed that the levels of ALT,AST,TC,TG,MDA contents,and CAT and GSH-Px activity of mice in the model group increased significantly（ P 〈 0. 01）,while the activity of SOD and weight and liver index decreased significantly（ P 〈 0. 01） compared with the normal control group. After 4 weeks of gavage,VCFP could significantly improve weight,liver index and SOD activity,and significantly reduce ALT,AST,TC and TG levels,MDA content and CAT and GSH-Px activity. These results suggest that VCFP can improve antioxidant enzyme activity,remove oxygen free radical and reduce membrane lipid peroxidation reaction,thereby protecting liver cells.

Staging of Adrenal Gland and Pancreas Neoplastic Cystwith Ultradiluted Viscum album:Case Report

Many animal adrenal lesions are frequently observed at necropsy and most often are asymptomatic with clinical syndromes that cause hypofunction or hyperfunction of the gland.The objective of this report is to demonstrate the staging of cysts and to report the clinical evolution of the dog that presented pathognomonic lesions of Cushing's Syndrome and developed Adrenal Cyst and a mass in the Pancreas.We concluded that the use of ultradiluted Viscum album in this patient promoted the control and staging of cystic tumor disease,pruritus and improved immunity to date,totaling 22 months of injectable therapy.

Viscum Album Modulates Apoptotic Related Genes in Melanoma Tumor of Mice

Cancer is a major public health problem throughout the world. It is estimated that one third of the American population will develop the disease at some time during their lifetimes. Among these, melanoma will account for 7% of the cases. In Brazil, in 2012, it is estimated that over six thousand new melanoma cases occurred. During recent years, the incidence of melanoma has increased, mainly due to a more constant exposure of the skin to sunlight. In this work, our aim is to assess the expression of apoptotis-related genes melanoma tumors in mice treated with Viscum album (VA). This will allow us to better understand the molecular mechanisms underlying tumor cell death activation caused by this compound. Our results clearly demonstrate upregulation of pro apoptotic genes (Trp53bp2, Nol3, Fadd, Tnfsf10, Traf1, Traf2, Cflar, Card10, Nod1, Casp 2, Casp7, Xiap, Dad1, and Dffb). Further bioinformatics-based tool analysis allowed us to assess which specific cell death-related intracellular pathways were activated by VA treatment. Two major effects of VA in melanoma cells could be observed: generation of an immunomudulatory Th-1 like action, recruiting several interleukines, and cell death activation through Caspase7, associated uspstream with Card10 and downstream with CAD. In summary, VA modulates apoptosis related genes in cancer melanoma cells. Although a deeper study should be conducted, VA seems to interfere with important signaling pathways within melanoma cells that control the cellular mechanisms of apoptosis activation. Therapeutic approaches using VA as an antineoplastic and adjuvant medication compounding should be considered.

Three new bioactive flavonoid glycosides from Viscum album

Two new flavonoid glycosides, named viscumneoside XⅡ(1), and viscumneoside XⅢ(2);a new dihydrogen flavonoid glycoside product named viscumneoside XⅣ(3), were isolated from the aerial part of Viscum album, along with seven known compounds(4-10). Their structures were identified by analysis of spectroscopic data. In addition, cytotoxicity assay showed that 1, 2 and 3 possessed significant inhibitory activities against C6, A549 and MDA-MB-231(the inhibition rate arrived about 50%, 70% and74% respectively with IC50 ≤ 60.00 μmol·L^-1), while the inhibition of TF-1 and Hela was not significant.

Factors shaping community structure of endophytic fungi–evidence from the Pinus-Viscum-system

Endophytic fungal communities have been shown to be highly diverse in almost every host plant species analyzed so far.However,the factors shaping their compositions are largely unknown.To elucidate the impact of various factors,10 independent replicates of DNA extracts from each of 17 surface-sterilized leaf and stem samples were analyzed by pyrosequencing of fungal ITS1 rRNA gene amplicons.Altogether,154 fungal OTUs(operational taxonomic units),represented by 953,385 sequences,were found in at least 2 samples from Viscum album ssp.austriacum and/or its host Pinus sylvestris.Deviating from earlier,cultivation-based assessments,the communities were dominated by OTUs related to the genus Mortierella and OTUs not assignable to a certain fungal phylum.However,Ascomycota were still the most diverse group in terms of OTU richness and already hypothesized organ and host preferences of certain endophytic Xylariaceae isolated from the Pinus-Viscum-system could be confirmed.Host species and organ type were also the major factors shaping the detected fungal communities.The two plant species clearly differed according to the endophytic fungal communities,but only stems and needles of Pinus were inhabited by significantly different fungal assemblages.Interestingly,only the 1 and 3 year old stem sections differed according to the endophytic fungal community,while differently aged leaves of both plants were indistinguishable in this regard.Size of the organs had no impact on fungal communities in Pinus,but shorter internodes and smaller leaves showed at least a tendency to differ from the corresponding larger organs in Viscum.Fungal communities also differed slightly between the two sampling sites,lying 200 km apart,and between the three sampling campaigns.Because the samples were drawn within 15 days,this finding indicates that seasonal shifts clearly outweigh aging effects in host plant with perennial leaves.The results therefore provide strong evidence against a linear development of the endophytic fungal communities in Pinus sylvestris and Viscum album over the years.The communities seem to establish themselves already in the year the respective organs emerge.Further study is required to clarify whether they predominantly establish anew each year,or if the core community persists throughout subsequent years.The most abundant endophytic OTUs are known from soil and/or dead plant material and are supposed to represent latent decomposers.The study reveals for the first time that host and/or organ preferences of putatively saprotrophic fungi are predominantly responsible for compositional differences in the endophytic fungal communities between host plants and organs.While the analyses are shown to provide rather robust results,the significance of genetic abundance,as revealed by high-throughput sequencing analyses,remains an unsettled issue.This is discussed in detail,as well as the challenges in assigning taxonomic names to OTUs.

Two symmetrical unsaturated acids isolated from Viscum album

In the present study,two new acetylene conjugate compounds,dibutyl(2Z,6Z)-octa-2,6-dien-4-yne dioate(1),and dibutyl(2E,6E)-octa-2,6-dien-4-yne dioate(2),were isolated from the dry stem leaves of Viscum album,along with nine known compounds(3–11).Their structures were confirmed on the basis of spectroscopic data.Compounds 1 and 8 showed antioxidant activity against xanthine oxidase(XOD)and 1,1-diphenyl-2-picrylhydrazyl radical 2,2-diphenyl-1-(2,4,6-trinitrophenyl)hydroxyl(DPPH),with the IC_(50) of 1.22 and 1.33μmol·L^(–1),and the SC_(50 )of 4.34 and 8.22μmol·L^(–1),respectively.

Study on the dynamic variation of the secondary metabolites in Viscum coloratum using targeted metabolomics

Viscum coloratum(Kom.)Nakai is a well-known medicinal plant.However,the optimal harvest time for V.coloratum is unknown.Few studies were performed to analyze compound variation during storage and to improve post-harvest quality control.Our study aimed to comprehensively evaluate the quality of V.coloratum in different growth stages,and determine the dynamic variation of metabolites.Ultra-performance liquid chromatography tandem mass spectrometry was used to quantify 29 compounds in V.coloratum harvested in six growth periods,and the associated biosynthetic pathways were explored.The accumulation of different types of compounds were analyzed based on their synthesis pathways.Grey relational analysis was used to evaluate the quality of v.coloratum across different months.The compound variation during storage was analyzed by a high-temperature high-humidity accelerated test.The results showed that the quality of V.coloratum was the hightest in March,followed by November,and became the lowest in July.During storage,compounds in downstream steps of the biosynthesis pathway were first degraded to produce the upstream compounds and some low-molecular-weight organic acids,leading to an increase followed by a decrease in the content of some compounds,and resulted in a large gap during the degradation time course among different compounds.Due to the rapid rate and large degree of degradation,five compounds were tentatively designated as“early warning components”for quality control.This report provides reference for better understanding the biosynthesis and degradation of metabolites in V.coloratum and lays a theoretical foundation for rational application of V.coloratum and better quality control of V.coloratum during storage.