Evaluation of Anti-anemic Effect of Anthocyanin Rich Oryza sativa L. indica (Black Rice) Seeds Extract

Oryza sativa L. indica (black rice) has a rich cultural history and used as a tribute food. As the outer layer of seeds contains anthocyanin, O. sativa L. indica is black. Traditionally the plant used to enhance heart health, reduces the risk of atherosclerosis, diabetes as well as allergy, control hypertension, and improves the digestive system and lipid profile. It has anti-inflammatory action, detoxifies the body, decreases cancer growth, supports in weight management, and improve memory and quality of life. Here, the methanolic extract of black rice (BRME) was evaluated for its anti-anemic effect. In study anemia was induced by administering 40 mg/kg phenylhydrazine (PHZ) for two consecutive days in rats. From day 3, animals were treated with standard drug and BRME extracts (200 and 400 mg/kg) till 15 days. The anti-anemic activity was examined by estimating different parameters like hemoglobin, red blood cell, hematocrit, reticulocytes, aspartate amino transaminase activity, alanine amino transaminase and alkaline phosphatase. The results of the phytochemical test revealed the presence of carbohydrates, alkaloids, glycosides, saponins, flavonoids, tannins, amino acid, and triterpenoids. Methanolic extract of black rice also restored the hematological parameter and biochemical parameters values near to normal levels after 13 days of treatment at 200 and 400 mg/kg. The 400 mg/kg dose shows higher anti-anemic effect compared to the lower 200 mg/kg dose. From the result, it is concluded that O. sativa L. indica is a valuable source of active compounds with anti-anemic properties and can be used to treat anemia.

Mechanism of Berberis atrocarpa anthocyanin against Alzheimer's disease based on network pharmacology and experimental verification / 中国中药杂志

This study aimed to explore the potential mechanism of Berberis atrocarpa Schneid. anthocyanin against Alzheimer's disease(AD) based on network pharmacology, molecular docking technology, and in vitro experiments. Databases were used to screen out the potential targets of the active components of B. atrocarpa and the targets related to AD. STRING database and Cytoscape 3.9.0 were adopted to construct a protein-protein interaction(PPI) network and carry out topological analysis of the common targets. Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment analyses were performed on the target using the DAVID 6.8 database. Molecular docking was conducted to the active components and targets related to the nuclear factor kappa B(NF-κB)/Toll-like receptor 4(TLR4) pathway. Finally, lipopolysaccharide(LPS) was used to induce BV2 cells to establish the model of AD neuroinflammation for in vitro experimental validation. In this study, 426 potential targets of active components of B. atrocarpa and 329 drug-disease common targets were obtained, and 14 key targets were screened out by PPI network. A total of 623 items and 112 items were obtained by GO functional enrichment analysis and KEGG pathway enrichment analysis, respectively. Molecular docking results showed that NF-κB, NF-κB inhibitor(IκB), TLR4, and myeloid differentiation primary response 88(MyD88) had good binding abilities to the active components, and malvidin-3-O-glucoside had the strongest binding ability. Compared with the model group, the concentration of nitric oxide(NO) decreased at different doses of malvidin-3-O-glucoside without affecting the cell survival rate. Meanwhile, malvidin-3-O-glucoside down-regulated the protein expressions of NF-κB, IκB, TLR4, and MyD88. This study uses network pharmacology and experimental verification to preliminarily reveal that B. atrocarpa anthocyanin can inhibit LPS-induced neuroinflammation by regulating the NF-κB/TLR4 signaling pathway, thereby achieving the effect against AD, which provides a theoretical basis for the study of its pharmacodynamic material basis and mechanism.

In vivo antioxidant activity of rabbiteye blueberry (Vaccinium ashei cv. 'Brightwell') anthocyanin extracts / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Blueberries are rich in phenolic compounds including anthocyanins which are closely related to biological health functions. The purpose of this study was to investigate the antioxidant activity of blueberry anthocyanins extracted from 'Brightwell' rabbiteye blueberries in mice. After one week of adaptation, C57BL/6J healthy male mice were divided into different groups that were administered with 100, 400, or 800 mg/kg blueberry anthocyanin extract (BAE), and sacrificed at different time points (0.1, 0.5, 1, 2, 4, 8, or 12 h). The plasma, eyeball, intestine, liver, and adipose tissues were collected to compare their antioxidant activity, including total antioxidant capacity (T-AOC), superoxide dismutase (SOD) activity and glutathione-peroxidase (GSH-PX/GPX) content, and the oxidative stress marker malondialdehyde (MDA) level. The results showed that blueberry anthocyanins had positive concentration-dependent antioxidant activity in vivo. The greater the concentration of BAE, the higher the T-AOC value, but the lower the MDA level. The enzyme activity of SOD, the content of GSH-PX, and messenger RNA (mRNA) levels of Cu,Zn-SOD, Mn-SOD, and GPX all confirmed that BAE played an antioxidant role after digestion in mice by improving their antioxidant defense. The in vivo antioxidant activity of BAE indicated that blueberry anthocyanins could be developed into functional foods or nutraceuticals with the aim of preventing or treating oxidative stress-related diseases.

Comparative metabolomics provides novel insights into the basis of petiole color differences in celery (Apiumgraveolens L.) / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Plant metabolites are important for plant development and human health. Plants of celery (Apiumgraveolens L.) with different-colored petioles have been formed in the course of long-term evolution. However, the composition, content distribution, and mechanisms of accumulation of metabolites in different-colored petioles remain elusive. Using ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS), 1159 metabolites, including 100 lipids, 72 organic acids and derivatives, 83 phenylpropanoids and polyketides, and several alkaloids and terpenoids, were quantified in four celery cultivars, each with a different petiole color. There were significant differences in the types and contents of metabolites in celery with different-colored petioles, with the most striking difference between green celery and purple celery, followed by white celery and green celery. Annotated analysis of metabolic pathways showed that the metabolites of the different-colored petioles were significantly enriched in biosynthetic pathways such as anthocyanin, flavonoid, and chlorophyll pathways, suggesting that these metabolic pathways may play a key role in determining petiole color in celery. The content of chlorophyll in green celery was significantly higher than that in other celery cultivars, yellow celery was rich in carotenoids, and the content of anthocyanin in purple celery was significantly higher than that in the other celery cultivars. The color of the celery petioles was significantly correlated with the content of related metabolites. Among the four celery cultivars, the metabolites of the anthocyanin biosynthesis pathway were enriched in purple celery. The results of quantitative real-time polymerase chain reaction (qRT-PCR) suggested that the differential expression of the chalcone synthase (CHS) gene in the anthocyanin biosynthesis pathway might affect the biosynthesis of anthocyanin in celery. In addition, HPLC analysis revealed that cyanidin is the main pigment in purple celery. This study explored the differences in the types and contents of metabolites in celery cultivars with different-colored petioles and identified key substances for color formation. The results provide a theoretical basis and technical support for genetic improvement of celery petiole color.

Promoting effects of Difrarel ? on retinal function following panretinal photocoagulation in diabetic retinopathy / 中华实验眼科杂志

Objective:To investigate the promoting effect of Difrarel ? on retinal function following panretinal photocoagulation (PRP) in the eyes with diabetic retinopathy. Methods:A non-randomized controlled study was performed.A total of 108 eyes of 108 patients with non-proliferative diabetic retinopathy (NPDR) were enrolled in Tongren Ophthalmology Center and Beijing Daxing District People's Hospital from December 2014 to February 2020.The patients were divided into PRP group and PRP+ Difrarel ? group according to different therapies under patients' selection.Difrarel ? was orally administered after PRP in 56 patients of PRP+ Difrarel ? group, and only PRP was given in 52 patients of PRP group.The visual acuity, 30°~60° circular visual field and multifocal electroretinogram were examined before and 1 day, 1 month, 3 months, 6 months, 12 months after PRP.The central macular thickness (CMT) was measured by optical coherence tomography, and fundus neovascularization was observed by fluorescein fundus angiography at 6 and 12 months after PRP.The study protocol was approved by an Ethics Committee of Beijing Daxing District People's Hospital (No.2021-F4). Results:Visual improvement rate was 57.14% (32/56) and 32.69% (17/52) in PRP+ Difrarel ? group and PRP group at the end of following-up, respectively, showing a significant difference between two groups ( χ2=3.56, P<0.05). The visual field mean sensitivity was significantly different at different time points in two groups ( Fgroup=4.77, P<0.05; Ftime=6.51, P<0.05), and was lower after PRP than those before treatment in both groups (both at P<0.05), and was significantly higher in PRP+ Difrarel ? group than PRP group at 3, 6, 12 months after PRP (all at P<0.05). The P1 amplitude density in 3 to 5 rings in PRP+ Difrarel ? group were higher than those in PRP group, and the differences were statistically significant (all at P<0.05). There was no significant difference in CMT between the two groups at different time points ( Fgroup=3.57, P>0.05; Ftime=1.23, P>0.05). No new blood vessels and non-perfusion area were found in both groups. Conclusions:Oral Difrarel ? can improve retinal function after PRP in the eyes with NPDR.

Research Progress on Active Ingredients of Flavonoids in Traditional Chinese Medicine Dendrobium / 中国实验方剂学杂志

Dendrobium（Orchidaceae） is traditional Chinese medicine with a high healthcare value，which can nourish Yin and tonify deficiency. Eighty-nine flavonoids were isolated from Dendrobium，mainly including flavones，flavanoes and flavonols. Among them，there were 40 flavonoids，the main aglycones were apigenin and chrysoeriol；20 flavanones；and 15 flavonols，and the main aglycones were kaempferol and quercetin. D. officinale and several other species also have flavanonols，anthocyanidins，chalcone and isoflavones. There were 34 species of Dendrobium containing flavonoids，including 38 flavonoids in D. officinale，28 flavonoids in D. huoshanense，19 flavonoids in D. devonianum，12 flavonoids in D. wardianum，5 flavonoids in D. thyrsiflorum，4 flavonoids in D. denneanum and D. findlayanum. Common flavonoids included naringenin，quercetin，rutin，which had pharmacological effects of resisting oxidation，lowering blood sugar，improving blood circulation，lowing cholesterol，and protecting the cardiovascular system. The existing studies of Dendrobium-related species，flavonoids and their physiological functions were reviewed in the expectation to promote the attention of the industry to Dendrobium flavonoids and explore values of Dendrobium plants in efficacy and food fields.

Gall traits and galling insect survival in a multi-enemy context / Características de las agallas, y supervivencia de insectos que producen agallas, en el contexto de enemigos múltiples

Introduction: The enemy hypothesis postulates that gall traits protect galling insects against natural enemies. Galls show a huge range of sizes, colors and ornaments, which vary even intraspecifically. However, galling insects are targets of various organisms that attack them directly or indirectly. In this context, to consider only one gall trait to investigate gall structure acting against only one guild of enemies can conceal an understanding of the community-level interactions. Objective: Herein, we take these ideas into consideration to investigate the conspicuous galls induced by Palaeomystella oligophaga Becker and Adamski 2008 (Lepidoptera) on Macairea radula (Bonpl.) (Melastomataceae) as a model system. We characterize this system through categorization of the different enemy guilds present in the community. We identified them to the lowest taxonomic level possible and determined the kind of interaction responsible for galling insects' deaths. Considering the enemy hypothesis and the selection of secondary characteristics, we also aimed to determine which of the multiple gall traits influence the survival success of galling insects in a multi-enemy context. Methods: We inspected galls and characterized the enemy guilds affecting the galling insect and the mortality rates produced by each one of them. Next, we tested whether the distinct gall traits measured (parenchyma thickness, color, projections) promote galling insect survival with respect to each enemy. Results: The mortality induced by indirect enemies (organisms that interact with gall tissues and can interact secondarily with galling insect) was 47.3 %, being higher than that caused by parasitoids and predators (31.5 %). Despite the gall's structural complexity, live galling insects showed the smallest occurrence (21.2 %). Parenchyma thickness was negatively related to Calliephialtes parasitoids, Gelechiidae cecidophages and predation signals. Conclusions: We demonstrated that the attacks to gall tissues by the cecidophages represented the highest threat to P. oligophaga survival, being higher than the mortality caused by direct enemies. That is, the gall traits were not as efficient as supposed to protect the galling insect from the attack of natural enemies. Nevertheless, we also demonstrated that parenchyma thickness can be negatively related to some organisms, especially direct enemies. Other traits hypothesized as defensive (e.g. projections, coloration) may simply play no role.

Introducción: La hipótesis del enemigo postula que las características de la agalla protegen al agallero contra los enemigos naturales. Las agallas muestran una gran variedad de tamaños, colores y adornos, que varían incluso de forma intraespecífica. Sin embargo, los agalleros son objetivos de varios organismos que los atacan directa o indirectamente. En este contexto, considerar solo una característica para investigar la estructura de la agalla actúando contra un solo gremio de enemigos puede ocultar una comprensión de las interacciones a nivel comunitario. Objetivos: Para investigar las ideas presentadas usamos las agallas conspicuas inducidas por Palaeomystella oligophaga Becker y Adamski 2008 (Lepidoptera) en Macairea radula (Bonpl.) (Meslastomataceae) como sistema modelo. Describimos este sistema a través de la categorización de los diferentes gremios enemigos presentes en la comunidad. Los identificamos al nivel taxonómico más bajo posible y determinamos el tipo de interacción responsable de la muerte de los agalleros. Teniendo en cuenta la hipótesis del enemigo y la selección de características secundarias, también buscamos determinar cuáles de las múltiples caracteristicas de la agalla influyen en el éxito de supervivencia de los agalleros en un contexto de enemigos múltiples. Métodos: Inspeccionamos las agallas y caracterizamos los gremios enemigos que afectan al agallero y las tasas de mortalidad producidas por cada uno de ellos. Luego, probamos si las distintas caracteristicas de las agallas medidas (grosor del parénquima, color, proyecciones) promueven la supervivencia de los agalleros con respecto a cada enemigo. Resultados: La mortalidad indirecta inducida por los cecidofagos fue del 47.3 %, superior a la causada por los parasitoides y los depredadores (31.5 %). Apesar de la complejidad estructural de la agalla, los agalleros vivos mostraron la menor presencia (21.2 %). El grosor del parénquima se relacionó negativamente con los parasitoides de Calliephialtes, los cecidófagos de Gelechiidae y la depredación. Conclusiones: Demostramos que los ataques a los tejidos biliares por cecidófagos representaron la mayor amenaza para la supervivencia de P. oligophaga. Esto es interesante porque, de acuerdo con la hipótesis del enemigo, la estructura de las agallas debería proporcionar protección para los insectos agalleros en lugar de atraer a los insectos que se alimentan de la agalla misma. Sin embargo, también demostramos que el grosor del parénquima puede estar relacionado negativamente con algunos organismos, especialmente los enemigos directos. Otras caracteristicas hipotéticas como defensivas (por ejemplo, proyecciones, coloración) pueden simplemente no desempeñar ningún papel.

Morphoagronomic and molecular characterization of Euterpe oleraceaaccessions from eastern Brazilian Amazon

Açaí (Euterpe oleraceaMart.) -a common tropical palm has high social, economic, and environmental importance in the Amazon region. In the light of increasing exploration to obtain the fruit and heart of this palms, comprehensive studies are warranted for conservation and genetic improvement. Here, we characterized açaí accessions using phenological, morphological, and agronomic descriptors and random amplified polymorphic DNA (RAPD)molecular markers for joint selection of accessions with greater productivity. Hundred accessions were analyzed using 18 morphoagronomic descriptors and 13 RAPD markers. The spathe and inflorescence emission phases during flowering and fruiting showed seasonality. Based on the coefficient of variation and mean squared error, the accessions exhibited high variability in the tested morphoagronomic descriptors and were distributed into seven groups. Fruit, seed, and pulp weights were important descriptors for the distinction of accessions and identification of those with greater productivity. The accessions presented >85% similarity, and 85 accessions, distributed in nine subgroups, could not be differentiated using RAPD markers. There was no correlation between grouping based on morphometric descriptors and RAPD markers. Panicle weight was 3.9-9.0 kg in 15 accessions and 100-fruit pulp weight was 35-50 g in six accessions. Therefore, accessions with high productivity could be selected.

Genetic analysis of some morphological traits in synthetic x naturally polyploid cotton derivatives

Cotton has received attention of geneticists since more than a century. Gossypium hirsutum, the predominantly cultivated cotton species worldwide, has a narrow genetic base. It is important to broaden its genetic base through introgression of novel alleles from related species. Here, we report the development and characterization of a backcross population derived from the hybridization of a ‘synthetic’ (derived by crossing and chromosomal doubling of nonprogenitor Gossypium species) and natural tetraploid upland cotton. ‘Synthetic’ was observed to be male-sterile and thus, was used as the female parent. A total of 7434 flowers were pollinated to obtain 1868 BC1F1 seeds by direct and reciprocal crosses. Characterization of the experimental plant material was conducted in the field for several morphological traits such as pubescence on the stem, leaf, petiole and bract, presence/absence of petal spot, petal margin colouration and stamen filament colouration. Genetic analysis revealed that petal margin colouration phenotype was governed by a single dominant gene, whereas the petal spot and filament colouration phenotypes manifested segregation distortion. None of the BC1F1 plants was devoid of trichomes thus demonstrating that presence of trichomes is dominant over their absence. Modern upland cotton cultivars are usually devoid of petal spot, petal margin colouration and stamen filament colouration. These floral anthocyanin pigmentation characteristics, if fixed in the cotton germplasm, may serve as diagnostic features for the identification of cultivars during DUS testing as well as in the maintenance breeding programmes

In Vitro Antioxidant Potentiality Of Purified Anthocyanin From Floral Petals Of Wild Balsam Species

Phenolics are the largest group of phytochemicals ubiquitous in plant species with considerable interest economically. Recently, search of novel polyphenols increasingly becomes an area of intensive pharmacological research due to their multiple bioactive features such as antioxidant, antimicrobial, anti-carcinogenic, anti-viral and anti-inflammatory potentialities. Anthocyanins are flavonoid group of polyphenols, a group predominant in flowers, fruits and vegetables. The flavonoids, perhaps the unique single group of phenolics in foods, comprise a group of over 4200 C15 aromatic molecules with multiple structural patterns. The functions of anthocyanins as medicinal have been well-accepted in folk medicine throughout the world. In fact, these molecules are connected to an amazingly broad-based range of health benefits. In this juncture, the aim of this work was to evaluate the antioxidant activities of purified anthocyanin from wild balsam species. Initially, anthocyanin was extracted from floral leaves of wild balsam species and purified by chromatographic techniques. Subsequently, it was subjected to NMR and LC MS analysis. The major fractions identified were hesperidin, dimethoxy antirrhinin and trimethoxy antirrhinin. Further, the anthocyanin extracts were subjected to in vitro protocols like 2,2’-azino-bis-3-ethylbenzothiazoline-6-sulphonic acid (ABTS) radical cation, DPPH scavenging assay, ferric reducing antioxidant power (FRAP), beta carotene bleaching assay, metal chelating and H2O2 scavenging power. Interestingly, ABTS, FRAP analyzes yielded significant results as compared to others. The data were comparable with that of synthetic antioxidants like ascorbate and catechin. Meanwhile, beta carotene and H2O2 scavenging assay showed moderate results. DPPH and metal chelating protocols displayed the values 71% and 64% respectively at 25 µg/ml concentration. This study provides model systems for the evaluation of natural antioxidants like anthocyanin. Future in vivo clinical studies are warranted to confirm the obtained data.

Mircowave-Assisted Extraction of Phytochemical Constituents in Roselle (Hibiscus sabdariffa L.)

Roselle (Hibiscus sabdariffaL.) is natural source of valuable components such as phenolics, flavonoids and anthocyanins. Anthocyaninis normally used in the food technology to replace synthetic pigments as well as medicinal properties due to antioxidants. Microwave irradiation is a feasible green solvent extraction method receiving great attention as it utilizes solvent at elevated temperature and controlled pressure conditions. Microwave extraction emerges as a promising inexpensive, simple and efficient technique. In our research, various variable like microwave power (20, 40, 60, 80, 100 W), microwave frequency (10, 20, 30, 40, 50 GHz), the micrwave irradiation duration (5, 10, 15, 20, 25 min), liquid to solid ratio (water: material, 2:1, 3:1, 4:1, 5:1, 6:1), extraction temperature (50, 55, 60, 65, 70C), number of extraction cycles (1, 2, 3, 4, 5) influencing to the microwave-assisted extraction of phytochemicals in roselle (Hibiscus sabdariffa L.)calyx were thoroughly examined. Total total phenolic (mg GAE/ 100g), total flavonoid (mg GE/ 100g), anthocyanin (mg/100g) were key indicators to define the optimal variable. Our resultsrevealed that microwave power at 80 W, frequency 40 GHz, duration 15 min, liquid to solid ratio 4:1, temperature 55C, 4 cycles of extraction were appropriate for extraction of phytochemical components inside roselle (Hibiscus sabdariffa L.)calyx

Progress in the genetic modification of blue flowers based on anthocyanin metabolism / 生物工程学报

As water-soluble, natural pigments, anthocyanins are responsible for the red, purple and blue colors of many flowers, which attract pollinators to spread pollen. The colors of flowers are also essential for plants to survive in the nature and become one of the most significant characteristics of ornamental plants. In the booming floriculture industry, to produce various flower colors could increase the richness of natural colors, but it is still difficult to breed flowers with coveted blue color. The diversity of flower color is mainly determined by the types and contents of anthocyanins and their derivatives. The synthesis of delphinidin pigments is the key factor for breeding blue flowers. However, there are no structural genes in many plants to biosynthesize delphinidin pigments. Blue flowers are successfully created by genetic engineering in recent years. In this paper, using common ornamental plants as examples, we review the mechanism of plant flower coloration from the aspects of the key factors affecting the synthesis of delphinidin pigment and the production strategies of blue flowers based on the regulation of anthocyanin metabolism. Different strategies of molecular breeding could provide opportunities to improve colors of other floriculture plants and to develop anthocyanin-rich economic crops, such as colored cotton with blue fibers.

Research progress on separation of natural anthocyanin by liquid-liquid partition chromatography and combination with liquid chromatography / 中草药

Liquid-liquid partition chromatography (LLC) is a continuous and efficient chromatography technique now widely used in the field of natural products. Natural anthocyanin is a flavonoid natural pigment with many physiological activities. Anthocyanin is very unstable and LLC has been widely used in the separation of natural anthocyanin because of its mild operating conditions and the avoidance of irreversible adsorption of samples. The research progress on separation of natural anthocyanin by LLC and combination with liquid chromatography from 2010 to 2019 is reviewed in this paper.

Research progress on chemical constituents and pharmacological activities of Lycium ruthenicum / 中草药

Lycium ruthenicum is a kind of medicinal and edible plant with excellent health-care effect, which is a unique medicinal plant in the desert region of northwest China. Phytochemical investigations have identified that the fruit of this herb contains a variety of bioactive ingredients, including anthocyanins, flavonoids, alkaloids, and polysaccharides, as well as of fatty acids, amino acids, and some trace elements (such as manganese, selenium, and zinc, etc). Modern pharmacological researches have demonstrated that both the extract of L. ruthenicum and its constituents exhibit a wide range of pharmacological activities, such as anti-oxidation, anti-fatigue, hypoglycemic and hypolipidemic activity, cardiovascular and liver protection, as well as immune-regulatory activity. The chemical constituents of L. ruthenicum and their pharmacological effects are systematically summarized in this paper, and all information presented here may strongly facilitate further investigations on the pharmacological activities of this herb and the development and applications of the related products of this herb.

Study on Protection Effect of Mulberry Anthocyanin- 3-glucoside on Epilepsy Model Mice and the Effect of Hippocampal BDNF/TrkB Pathway / 中国药房

OBJECTIVE：To study the protection ef fects of mulberry anthocyanin- 3-glucoside on epilepsy model mice and the effect of hippocampal brain derived neurotrophic factor （BDNF）/tyrosine kinase B （TrkB）pathway. METHODS ：Totally 120 C57BL/6 mice were randomly divided into normal group ，model group ，single medication group （mulberry anthocyanin- 3- glucoside），agonist combination group（mulberry anthocyanin- 3-glucoside+TrkB agonist LM 22B-10），with 30 mice in each group. single medication group and agonist combination group were given mulberry anthocyanin- 3-glucoside 600 μg/kg intragastrically once a day ，for consecutive 6 weeks. The agonist combination group was given LM22B-10（5 mg/kg）via the lateral ventricle once a day at 6th week. Normal group and model group were given constant volume of normal saline intragastrically. After last medication，except for normal group ，other groups were given lithium chloride-pilocarpine to establish epilepsy model. After modeling，10 mice in each group were taken to record the latency ，frequency and duration of spontaneous recurrent epilepsy ， observed for 6 hours a day for 4 weeks；EEG was recorded on the 14th，28th and 36th day after modeling ，and the abnormal frequency of EEG in 1 h was counted . On the 6th day of modeling ，other 10 mice in each group were taken to detect the serum calcium level ，and the remaining 10 mice in each group were taken to detect the expressions of BDNF mRNA and protein in the hippocampus. RESULTS ：Compared with normal group ，latency，frequency and duration of spontaneous recurrent epilepsy and the times of abnormal brain wave on the 14th，28th and 36th day after modeling were increased significantly in model group （P＜ 0.05）. The serum calcium level ， mRNA and proteinexpression of BDNF in hippocampus were increased E-mail：wangfang7699@126.com significantly （P＜0.05）. Compared with model group ，the latency，frequency，duration of spontaneous recurrent epilepsy and the times of abnormal brain wave on the 28th and 36th day after modeling were decreased significantly in single medication group（P＜0.05），while serum calcium level ，mRNA and protein expression of BDNF in hippocampus were decreased significantly （P＜0.05）. Compared with single medication group ，the latency，frequency and duration of spontaneous recurrent epilepsy and the times of abnormal brain wave on the 28th and 36th day after modeling were increased significantly in agonist combination group （P＜0.05），while serum calcium level ，mRNA and protein expressions of BDNF in hippocampus were increased significantly （P＜0.05）. CONCLUSIONS ：Mulberry anthocyanin- 3- glucoside has a good protection effect on epilepsy model mice ，the mechanism of which may be associated with inhibiting the activation of hippocampal BDNF/TrkB pathway.

Exploring the binding affinity and non-covalent interactions of anthocyanins with aging-related enzymes through molecular docking

Background and Objective@#Anthocyanins are associated with aging and longevity. However, the mechanism involving the pure anthocyanin compounds in aging remains elusive. To investigate the possible mechanism of action of the different anthocyanin compounds towards aging-associated enzymes, the lead-likeness, binding affinity, and binding interactions were evaluated.@*Methodology@#The different anthocyanin compounds such as cyanidin, delphinidin, malvidin, pelargonidin, peonidin, and petunidin were assessed for lead-likeness following the criteria of Lipinski's rule of five (Ro5). These same compounds were virtually docked to different aging-related enzymes involved in MAPK, AMPK, and insulin signaling pathways. The top binding anthocyanins for each enzyme were visualized and compared to the enzyme inhibitors. @*Results@#The different anthocyanin compounds abide with Ro5 denoting its potential as a lead compound. For each enzyme, there were different top-binding anthocyanins. The crystal structures of the docked anthocyanins reveal that there were different substructures involved during the non-covalent interaction. Some substructures, particularly the hydroxy groups, have different roles during the H-bond formation. These findings suggest that the various anthocyanin compounds may have a distinct mechanism of action towards a specific enzyme. @*Conclusion@#Taken together, these results suggest that the anthocyanin compounds may have varying effects in aging enzymes, which may be due to the differences in their substructures. Nonetheless, further investigations are needed to understand these findings using cells and animal models.

Changes of pigment components in Dendrobium catenatum flower under different storage conditions / 中国中药杂志

The flower color of Dendrobium catenatum(D. officinale) tends to fade during storage. In order to clarify the influence of storage conditions on the pigment components in flowers, two conditions were applied:temperature and illumination. The contents of pigments in the D. catenatum flower were determined by UV-Vis spectrophotometry and HPLC, and the changes of them during storage were analyzed. The results showed that illumination and temperature had an effect on the pigments of D. catenatum flower during sto-rage. Illumination significantly promoted the degradation of pigments. The contents of total chlorophyll, carotenoids and anthocyanins in the light samples were significantly lower than those in the dark. The total chlorophyll, carotenoids and anthocyanins in the light samples were decreased by 46.5%, 63.4%, and 69.2% respectively. Illumination had a greater effect on fat-soluble pigments than water-soluble pigments. Among the three temperature treatments, the contents of chlorophyll, carotenoid and anthocyanin were as follows:-20 ℃>4 ℃>room temperature, it is indicated that-20 ℃ was the best temperature to maintain the stability of pigment composition. The contents of chlorophyll a, chlorophyll b, β-carotene, lutein and zeaxanthin in the light samples decreased by 34.8%, 69.0%, 72.5%, 61.6%, 36.1%, respectively. After storage for 5 months, the contents of chlorophyll, carotenoid and anthocyanin constituent at-20 ℃ was significantly higher than those at 4 ℃ and room temperature. The results show that light avoiding and low-temperature can effectively slow down the degradation of pigment components. Therefore, it is suggested that D. catenatum flower should be stored in light avoiding and low-temperature conditions in actual production and processing, which can prolong the usable time.

Molecular characterization of PgUFGT gene and R2R3-PgMYB transcription factor involved in flavonoid biosynthesis in four tissues of wild pomegranate (Punica granatum L.)

The diversity on fruit colouration in plants directly depends on the flavonoids that explain the development of different pigmentation patterns. Anthocyanins are the major class of flavonoid pigments that are synthesized through flavonoid biosynthetic pathway. In the present study, two genes: PgUFGT gene and R2R3-PgMYB gene, involved in anthocyanin biosynthesis were analysed in four tissues of wild pomegranate. The structural genes, UDP-glucose: flavonoid-3-O-glucosyl transferase (PgUFGT; GenBank accession number: MK058491) and its myeloblastosis transcription factor (R2R3-PgMYB; GenBank accession number: MK092063) were isolated and their expression pattern were studied. Molecular modelling indicated that the main secondary structures of PgUFGT and R2R3-PgMYB genes are α-helix and random coil. In addition, expression profiling of PgUFGTand R2R3-PgMYB by quantitative-real time PCR indicated a positive correlation between anthocyanin content and their expression in leaves, flowers, green and red fruits of wild pomegranate. Among all the tissues, the red fruit exhibited high transcripts levels of PgUFGT as well as R2R3-PgMYB transcription factor. An extensive homology with UFGTs from other plants was revealed on comparative and bioinformatic analyses. Present study reveals that PgUFGT plays a predominant role in anthocyanin content in wild pomegranate fruits. Further, it is strongly suggested that R2R3-PgMYB transcription factor regulates the anthocyanin biosynthesis in wild pomegranate via expression of PgUFGT gene. This is the first study which provides an insight on expression profile of PgUFGT and R2R3-PgMYB that are involved in colour development and fruit ripening in wild pomegranate.

Effect of intravitreal and intraperitoneal cyanidin-3-glucoside injection in oxygen-induced retinopathy mouse model

Purpose: To evaluate the effect of cyanidin-3-glucoside (C3G) in oxygen-induced retinopathy (OIR) mouse model. Methods: In this experimental study, 10 C57BL / 6J type mice exposed to room air comprised two control groups (n = 5 each; a negative control and a group receiving intravitreal sterile dimethyl sulfoxide [IVS DMSO]). Thirty C57BL / 6J type mice exposed to 75% ± 2% oxygen from postnatal day 7 to postnatal day 12 comprised the OIR groups. On postnatal day 12, these mice were randomized into six groups (n = 5 each): two OIR control groups (negative control and IVS DMSO), two intravitreal C3G groups (300 and 600 ng/?L), and two intraperitoneal C3G groups (0.05 and 0.1 mg/kg). We quantified neovascularization by counting endothelial cell proliferation on the vitreal side of the inner limiting membrane of the retina and examined histological and ultrastructural changes via light and electron microscopy and apoptosis by terminal deoxynucleotidyl transferase deoxy-UTP-nick end labeling. Results: The intravitreal C3G groups yielded lower endothelial cell counts compared with the intravitreal DMSO group. The intraperitoneal high-dose group had lower cell counts compared with the OIR control groups. Electron microscopy revealed significantly less mitochondrial dysmorphology in intravitreal groups and the high-dose intraperitoneal mice. We noted no difference in apoptotic cell count between the controls, low-dose intravitreal, and both intraperitoneal groups. However, apoptotic cell count was significantly higher in the high-dose intravitreal group. Conclusion: C3G suppresses endothelial cell proliferation in an OIR mouse model, leads to a reduced hyperoxia-induced mitochondrial dysmorphology, but increases apoptotic cell death in high concentrations.

Effects of black chokeberry extracts on metastasis and cell-cycle arrest in SK-Hep1 human liver cancer cell line

Objective: To determine the anti-cancer properties of black chokeberry extract on the SK- Hep1 human liver cancer cell line. Methods: MTT cell proliferation assay, wound migration, invasion, zymography and cell cycle were determined after black chokeberry fruit extract treatment. We also measured MMP-2/-9 and MT-1 MMP expression with protein and gene expression levels. Results: We detected four anthocyanins and three phenolic compounds in the black chokeberry by HPLC analysis. Cancer cell growth was inhibited in proportion to the concentration of black chokeberry extracts. In the adhesion test, 100 and 200 μg/mL of black chokeberry extracts decreased the adhesion rate of cancer cells to 87.6% and 75.3%, respectively, when the control group was 100.0%. The 200 μg/mL of black chokeberry extract reduced the MMP-2 and MMP-9 expressions up to 96.8%and 11.3%, respectively. Conclusions: Based on our results, in the SK-Hep1 liver cancer cells, the black chokeberry extract inhibits cancer cell proliferation, adhesion, and migration, ultimately inhibiting cancer metastasis.