Ameliorative effect of ferruginol on isoprenaline hydrochloride-induced myocardial infarction in rats.

Cardiovascular-related diseases continue to be a leading cause of death globally. Among ischemic-induced cardiac diseases, myocardial infarction (MI) is reported to be of an alarming value. Despite numerous improvements in the medical intrusions, still this armamentarium fails to be effective in managing the illness without setbacks. Ferruginol (FGL) is a major polyphenols and terpenoids with numerous pharmacological activities including antioxidant and anti-inflammatory. Following, this work was aimed to explore the cardio protective effect of FGL (50 mg/kg) in isoprenaline hydrochloride (ISO)-induced MI in experimental rats. After treatment with FGL in ISO-induced MI in rats, noticeable changes were observed in the experimental rats. Injection of ISO to rats resulted in the augmented cardiac weight, serum cardiac markers (creatine kinase, creatine kinase-MB, cardiac troponin T, and Cardiac troponin I), lipid peroxidation end products (thiobarbituric acid-reactive substance and lipid hydroperoxides), reduced endogenous antioxidants (superoxide dismutase, catalase, glutathione peroxidase, and glutathione), reduced ATPase activity, and escalated pro-inflammatory cytokines (interleukin-6, tumor necrosis factor-α, and nuclear factor-κB) levels. Interestingly, the FGL supplementation to the ISO-treated rats revealed the diminished heart weight, reduced cardiac markers, and lipid peroxidation. FGL also possessed the improved antioxidants status and diminished pro-inflammatory mediator levels. The outcomes of histological analysis also evidenced the cardio protective role of FGL. Treatment with FGL reduced the cardiac damage biomarkers maintained to near normal levels in ISO-induced rats. These study findings disclose the prospective capability of FGL in the treatment of MI in the future.

Transferrin improved the generation of cardiomyocyte from human pluripotent stem cells for myocardial infarction repair.

Human pluripotent stem cell (hPSC)-derived cardiomyocytes (CMs) hold great promise for the repair of the injured heart, but optimal cell production in a fully chemically defined and cost-effective system is essential for the efficacy and safety of cell transplantation therapies. In this study, we provided a simple and efficient strategy for cardiac differentiation from hPSCs and performed functional evaluation in a rat model of myocardial infarction. Using a chemically defined medium including four components, recombinant human albumin, ascorbic acid, human transferrin, and RPMI 1640, we developed a manageable and cost-effective protocol for robust generation of CMs from hPSCs. Interestingly, the addition of transferrin helped hPSCs to transit from TeSR-E8 medium to the simple cardiac differentiation medium and successfully initiated mesoderm differentiation without significant cell death. The CM generation efficiency was up to 85% based on cTnT expression. We performed transcriptome profiling from differentiation day 0 to 35, and characterized interesting dynamic change of cardiac genes. CMs derived from transferrin-supplemented simple medium have similar transcriptome and the maturation level compared to those generated in B27 minus insulin medium as well as their in vivo counterparts. Importantly, after transplantation, hPSC-derived CMs survived in the infarcted rat heart, significantly improved the physiological function and reduced fibrosis. Our study offers an easy-to-use and cost-effective method for cardiac differentiation and facilitates the translational application of hPSC-derived CMs for heart repair.

Anticancer Properties of Different Solvent Extracts of Cucumis melo L. Seeds and Whole Fruit and Their Metabolite Profiling Using HPLC and GC-MS.

Honeydew melon (Cucumis melo L.) is an oval-shaped delicious fruit of one cultivar group of the muskmelon with immense nutritional importance and is extensively consumed by many tropical countries. The effect of various organic solvents on the recovery of phytochemicals from honeydew melon plant fruits and seeds was assessed. Further, High-Performance Liquid Chromatography (HPLC) was used to examine and assess the contents of phenolic acid (gallic acid) and flavonoid (rutin) compounds. The use of gas chromatography-mass spectrometry (GC-MS) analysis explained the presence of volatile phytocompounds in the extracts. The use of organic solvents had a substantial impact on the total dry weight and extract yield. In general, the solvent-extracted constituents remained in the order of methanol>chloroform>distilled water for both honeydew melon seeds and whole fruit. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) was used to assess the cytotoxicity effect against PC3, HCT116, HeLa, and Jurkat cell lines. The chloroform extract exhibited a good cytotoxic activity against all cell lines as compared to other solvent extracts. HPLC analysis revealed the occurrence of gallic acid content of 0.102 ± 0.23 mg/10 mg of dry whole fruit extract, while 10 mg of dry seed extract contained only 0.022 ± 0.12 mg of gallic acid content. Likewise, rutin content was observed to be 0.224 ± 0.31 mg and 0.1916 ± 0.82 mg/10 mg of dry whole fruit and seed extract, respectively. Further, GC-MS analysis revealed the presence of a total of 37 compounds in chloroform extract of whole fruit, while only 14 compounds were found in seed extract. Nevertheless, more examinations are needed to identify and characterize other metabolites from honeydew melon and evaluate their pharmacological importance.

Anticancer and Antibacterial Activities of Silver Nanoparticles (AgNPs) Synthesized from Cucumis melo L.

The current investigation reports the structural and biological evaluation of silver nanoparticles (AgNPs) biosynthesized from the pericarp extract of Cucumis melo L. (muskmelon). The AgNPs were characterized by ultraviolet-visible (UV-Vis) spectrophotometry, XRD (X-ray diffraction), SEM (scanning electron microscopy) and EDAX (energy-dispersive X-ray spectroscopy). The XRD analysis showed that biosynthesized AgNPs were having FCC (face centered cubic) crystalline structures. Further, the SEM and EDAX showed spherically shaped AgNPs having an average size of 25 nm. The AgNPs effectively inhibited the growth of Bacillus subtilis and Escherichia coli. Moreover, the cytotoxic assay of AgNPs revealed effective cytotoxicity against different cancer cells, such as HeLa, HCT-116, PC-3 and Jurkat in a dose reliant way. The cell viability was noticed to range from 50% to 60% with IC50 values ranging from 150 µg/mL to 224 µg/mL. The lower cell viability indicates the toxic effects of biosynthesized AgNPs against these malignant cells. Thus, the current study shows that these biosynthesized AgNPs could be utilized in various medical applications in near future.

Eight new bibenzyl derivatives from Dendrobium candidum.

Eight bibenzyl derivatives, namely dendrocandins J-Q (1-8), were isolated from the stems of Dendrobium candidum. Their structures were elucidated by 1D and 2D NMR experiments and mass spectrometry. Compounds 1-8 were examined for antioxidant activity by 1,1-diphenyl-2-picrylhydrazyl free radical scavenging assay, and the IC50 values were 36.8, 70.2, 45.0, 60.5, 87.6, 50.4, 22.3, and 30.3 µM, respectively.

Stimulation of dragon's blood accumulation in Dracaena cambodiana via fungal inoculation.

Dragon's blood is a rare and precious traditional medicine used by different cultures since ancient times. However, studies on enhancing the rapid accumulation of dragon's blood in Dracaena cambodiana and determining its formation mechanism are unavailable. In this study, the activities of two fungi, namely, BJDC01 and BJDC05, and their effect on promoting the accumulation of five main compositions of dragon's blood in D. cambodiana were investigated for the first time. Results of field tests conducted for ten months indicated that the contents of Loureirin D, 4,4'-dihydroxy-2'-dimethoxychalcone, Loureirin A and Loureirin B in two fungal-inoculated materials were 1.67 to 2.85 times greater than those of natural samples, and thus were significantly higher than those of the control groups. The content of 4,4'-dihydroxy-2'6'-dimethoxydihydrochalcone in each fungal-inoculated sample was close to that of the natural sample, and was more than twice of each of the control group. By combining the results of morphological characterizations, both BJDC01 and BJDC05 can stimulate the accumulation of the compositions of dragon's blood. This stimulation may be considered as a defense response of D. cambodiana tree against the invasion of foreign fungi. Thus, this study provides a potential way of producing dragon's blood via the inoculation of two fungal elicitors.

Sclerotial formation of Polyporus umbellatus by low temperature treatment under artificial conditions.

BACKGROUND: Polyporus umbellatus sclerotia have been used as a diuretic agent in China for over two thousand years. A shortage of the natural P. umbellatus has prompted researchers to induce sclerotial formation in the laboratory. METHODOLOGY/PRINCIPAL FINDING: P. umbellatus cultivation in a sawdust-based substrate was investigated to evaluate the effect of low temperature conditions on sclerotial formation. A phenol-sulfuric acid method was employed to determine the polysaccharide content of wild P. umbellatus sclerotia and mycelia and sclerotia grown in low-temperature treatments. In addition, reactive oxygen species (ROS) content, expressed as the fluorescence intensity of mycelia during sclerotial differentiation was determined. Analysis of ROS generation and sclerotial formation in mycelia after treatment with the antioxidants such as diphenyleneiodonium chloride (DPI), apocynin (Apo), or vitamin C were studied. Furthermore, macroscopic and microscopic characteristics of sclerotial differentiation were observed. Sclerotia were not induced by continuous cultivation at 25°C. The polysaccharide content of the artificial sclerotia is 78% of that of wild sclerotia. In the low-temperature treatment group, the fluorescent intensity of ROS was higher than that of the room temperature (25°C) group which did not induce sclerotial formation all through the cultivation. The antioxidants DPI and Apo reduced ROS levels and did not induce sclerotial formation. Although the concentration-dependent effects of vitamin C (5-15 mg mL(-1)) also reduced ROS generation and inhibited sclerotial formation, using a low concentration of vitamin C (1 mg mL(-1)) successfully induced sclerotial differentiation and increased ROS production. CONCLUSIONS/SIGNIFICANCE: Exposure to low temperatures induced P. umbellatus sclerotial morphogenesis during cultivation. Low temperature treatment enhanced ROS in mycelia, which may be important in triggering sclerotial differentiation in P. umbellatus. Moreover, the application of antioxidants impaired ROS generation and inhibited sclerotial formation. Our findings may help to provide new insights into the biological mechanisms underlying sclerotial morphogenesis in P. umbellatus.

Discrimination of the rare medicinal plant Dendrobium officinale based on naringenin, bibenzyl, and polysaccharides.

The aim of this study was to establish a method for discriminating Dendrobium officinale from four of its close relatives Dendrobium chrysanthum, Dendrobium crystallinum, Dendrobium aphyllum and Dendrobium devonianum based on chemical composition analysis. We analyzed 62 samples of 24 Dendrobium species. High performance liquid chromatography analysis confirmed that the four low molecular weight compounds 4',5,7-trihydroxyflavanone (naringenin), 3,4-dihydroxy-4',5-dime-thoxybibenzyl (DDB-2), 3',4-dihydroxy-3,5'-dimethoxybibenzyl (gigantol), and 4,4'-dihydroxy-3,3',5-trimethoxybibenzy (moscatilin), were common in the genus. The phenol-sulfuric acid method was used to quantify polysaccharides, and the monosaccharide composition of the polysaccharides was determined by gas chromatography. Stepwise discriminant analysis was used to differentiate among the five closely related species based on the chemical composition analysis. This proved to be a simple and accurate approach for discriminating among these species. The results also showed that the polysaccharide content, the amounts of the four low molecular weight compounds, and the mannose to glucose ratio, were important factors for species discriminant. Therefore, we propose that a chemical analysis based on quantification of naringenin, bibenzyl, and polysaccharides is effective for identifying D. officinale.

Antimicrobial activities of endophytic fungi isolated from Ophiopogon japonicus (Liliaceae).

BACKGROUND: Drug resistance in bacteria has become a global concern and the search for new antibacterial agents is urgent and ongoing. Endophytes provide an abundant reservoir of bioactive metabolites for medicinal exploitation, and an increasing number of novel compounds are being isolated from endophytic fungi. Ophiopogon japonicus, containing compounds with antibacterial activity, is a traditional Chinese medicinal plant used for eliminating phlegm, relieving coughs, latent heat in the lungs, and alleviating diabetes mellitus. We investigated the antimicrobial activities of 30 strains of O. japonicus. METHODS: Fungal endophytes were isolated from roots and stems of O. japonicus collected from Chongqing City, southwestern China. Mycelial extracts (MC) and fermentation broth (FB) were tested for antimicrobial activity using peptide deformylase (PDF) inhibition fluorescence assays and MTT cell proliferation assays. RESULTS: A total of 30 endophytic strains were isolated from O. japonicus; 22 from roots and eight from stems. 53.33% of the mycelial extracts (MC) and 33.33% of the fermentation broths (FB) displayed potent inhibition of PDF. 80% of MC and 33.33% of FB significantly inhibited Staphylococcus aureus. 70% of MC and 36.67% of FB showed strong activities against Cryptococcus neoformans. None showed influence on Escherichia coli. CONCLUSION: The secondary metabolites of endophytic fungi from O. japonicus are potential antimicrobial agents.

[Fluid suspension co-culture of Dendrobium officinale protocorm and living fungus].

OBJECTIVE: To set up a system for fluid suspension co-culture of Dendrobium officinale protocorm and living fungus MF24, so as to provide certain scientific evidence for industrial production of protocorm. METHOD: Whether the protocorm culture system was suitable for the normal growth of MF24 fungus were studied, the growth of protocorm cultured alone and co-cultured with the fungus were researched under light and dark culture conditions, the biomass and proliferation times were determined, and HPLC method was used to analyze and compare the changes of 11 characteristic peak areas in D. officinale protocorm. RESULT: The MF24 fungus could grow normally in the 6,7-V liquid medium used to culture the protocorm, and when it was cultured by 8-10 hours per day under 1 500 lx, the growth rate of the fungus was slowed. Protocorm could grow normally in light and dark culture conditions, and add the MF24 fungus in the early cultivation stages of protocorm, both inhibit the growth of each other. In the protocorm for the growth stability to add 5 diameter 9 mm fungi block, the protocorm growth and chemical composition type had no significant effect. However, under illumination, co-cultured for 5 days protocorm of which 10 compounds content decreased 13.64% to 138.47%, in dark conditions, co-cultured for 5 days protocorm of which 7 compounds increased by 0.71% to 12.82%, and 4 compounds slightly reduced by 3.03% to 14. 14% compared with the control. CONCLUSION: Under the appropriate condition, living fungus MF24 could co-culture with the D. officinale protocorm, and affected the latter's secondary metabolite levels.

Simultaneous structural identification of natural products in fractions of crude extract of the rare endangered plant Anoectochilus roxburghii using H NMR/RRLC-MS parallel dynamic spectroscopy.

Nuclear magnetic resonance/liquid chromatography-mass spectroscopy parallel dynamic spectroscopy (NMR/LC-MS PDS) is a method aimed at the simultaneous structural identification of natural products in complex mixtures. In this study, the method is illustrated with respect to (1)H NMR and rapid resolution liquid chromatography-mass spectroscopy (RRLC-MS) data, acquired from the crude extract of Anoectochilus roxburghii, which was separated into a series of fractions with the concentration of constituent dynamic variation using reversed-phase preparative chromatography. Through fraction ranges and intensity changing profiles in (1)H NMR/RRLC-MS PDS spectrum, (1)H NMR and the extracted ion chromatogram (XIC) signals deriving from the same individual constituent, were correlated due to the signal amplitude co-variation resulting from the concentration variation of constituents in a series of incompletely separated fractions. 1H NMR/RRLC-MS PDS was then successfully used to identify three types of natural products, including eight flavonoids, four organic acids and p-hydroxybenzaldehyde, five of which have not previously been reported in Anoectochilus roxburghii. In addition, two groups of co-eluted compounds were successfully identified. The results prove that this approach should be of benefit in the unequivocal structural determination of a variety of classes of compounds from extremely complex mixtures, such as herbs and biological samples, which will lead to improved efficiency in the identification of new potential lead compounds.

[Chemical constituents of Dendrobium candidum].

The column chromatography on silica gel, sephadex LH-20 preparative HPLC were used to separate and purify the compounds from the stems of Dendrobium candidum. Twenty compounds were isolated and identified as 3,4'-dihydroxy-5-methoxybibenzyl(1), dihydroresveratrol(2), dendromoniliside E(3), denbinobin(4),2,4,7-trihydroxy-9, 10-dihydrophenanthrene(5), aduncin(6), (-)-loliolide(7), adenosine(8), uridine(9), guanosine(10), sucrose(11), 5-hydroxymethyl-furaldehyde(12), n-octacostyl ferulate(13), defuscin(14), n-triacontyl cis-p-coumarate(15), daucosterol(16), beta-sitosterol(17), hexadecanoic acid(18), hentriacontane(19), and heptadecanoic acid(20). Their structures were elucidated on the basis of spectroscopic data and physicochemical properties. All of the compounds were isolated from this plant for the first time.

Complete assignments of (1)H and (13)C NMR data of three new dihydrophenanthrofurans from Pleione yunnanensis.

Three new dihydrophenanthrofurans, pleionesins A-C (1-3), together with two known dihydrophenanthrenes (4-5) were isolated from the tubers of Pleione yunnanensis (Rolfe). The complete (1)H and (13)C NMR spectra assignments of these compounds were carried out using 1D and 2D NMR experiments ((1)H, (13)C, selective 1D NOE, HSQC and HMBC).

[Comparison of rutin and syringin content between tissue culturing seedlings and botanical drug of Saussurea involucrata].

OBJECTIVE: To compare the rutin and syringin content in tissue culturing seedlings and in botanical drug of Saussurea involucrata. METHOD: The HPLC with Hydro-RP C18 (4.6 mm x 250 mm, 5 microm) column was used, a mixture of acetonitrile-water (5:95) was used as a mobile phase, with flow rate of 1 mL x min(-1), column temperature at 25 degrees C and detection wavelength at 220 nm. RESULT: The effective constituents of tissue culturing seedlings were almost similar to the botanical drug. And syringin in tissue culturing seedlings was increased 4.35 times. CONCLUSION: It has a good prospect to acquire high-quality S. involucrata by tissue culturing seedlings.

Quantitative determination of five glucosyloxybenzyl 2-isobutylmalates in the tubers of Gymnadenia conopsea and Coeloglossum viride var. bracteatum by HPLC.

Glucosyloxybenzyl 2-isobutylmalates are one group of important active constituents in the tubers of Gymnadenia conopsea R. Br. and Coeloglossum viride (L.) Hartm. var. bracteatum (Willd.). For the purpose of quality evaluation of these two Chinese herbal medicines, it is necessary to use a rapid and reliable assay that is suitable for the determination of their active constituents. A high-performance liquid chromatography method is firstly developed for the simultaneous quantification of five glucosyloxybenzyl 2-isobutylmalates in the tubers. The analytes including dactylorhin B, dactylorhin E, loroglossin, dactylorhin A, and militarine are isolated from the tubers of G. conopsea. The compounds are separated on an Agilent Hydrosphere C(18) (150 x 4.6 mm i.d., 5 microm) column using a mobile phase of acetonitrile-water including 0.3% acetic acid (adjusted with 36% acetic acid) with gradient elution at a flow rate of 1.0 mL/min. Detection is set at a UV wavelength of 221.5 nm. The recovery of the method is 97.7-101.0%, and linearity (r > 0.9998) is obtained for all the analytes. The assay is successfully applied to determine the contents of the analytes in the tubers of G. conopsea and C. viride var. bracteatum collected from different regions of China.

New bibenzyl derivatives from the tubers of Pleione yunnanensis.

Four new bibenzyl derivatives, shancigusins A-D (1-4) and five known bibenzyls (5-9) were isolated from the tubers of Pleione yunnanensis (Orchidaceae). The structures of these compounds were determined by extensive analyses of their spectroscopic data.

Three guaianolides from Saussurea involucrata and their contents determination by HPLC.

Saussurea involucrata (Kar. et Kir.) Sch. Bip. (family Asteraceae) is a rare traditional Chinese medicinal herb. A new guaianolide, 11 beta,13-dihydrodehydrocostuslactone-8-O-[6'-O-acetyl-beta-D-glucoside] (1), together with two known guaianolides, namely 11 beta,13-dihydrodehydrocostuslactone-8-O-beta-D-glucoside (2) and 3 alpha-hydroxyl-11 beta,13-dihydrodehydrocostuslactone-8-O-beta-D-glucoside (3), was isolated from the 95% ethanol extract of the whole plants. Their structures were elucidated on the basis of spectral analysis. Their contents in the S. involucrata were determined by HPLC for the first time. Chromatographic separation was achieved on an Agilent Zorbax SB-C(18) column (250 mm x 4.6mm i.d., 5 microm) at 25 degrees C and the mobile phase was a mixture of acetonitrile and water in a mode of gradient elution detected at 209.8 nm at a flow rate of 1.0 ml/min. This method was validated in terms of selectivity, linearity, precision and accuracy. The linearity ranges were 0.066-0.44 microg for 1, 0.23-2.30 microg for 2, and 0.258-2.58 microg for 3, respectively.

[Advances in studies on chemical components and pharmacology of epiphytic type medicinal plants in the Orohid family].

Many of the epiphytic Orchids are used as traditional Chinese medicine. The chemical components and pharmacology have been studied in recent 15 years. This article reviewed the studies which will be beneficial to reveal the relatives among these medicinal plants in the Orchid Family and be helpful to develop new drugs.

[Studies on chemical components and pharmacological activities of geobiontic type medicinal plants in Orchidaceae family].

Geobiontic type medicinal plants subjected to Orchidaceae family are used as important traditional Chinese medicine. Scientists are paying more attentions to their chemical components and pharmacology recently. This paper reviewed their chemical components involved in phenanthrene, bibenzene, flavone, sterol, terpenes, alkoloids and pharmacology such as antibacterium, cytotoxic activity, antihyperliposis effects. All these information are beneficial to reveal the relative among these medicinal plants in Orchidaceae family and are helpful to develop new drugs.

[Study on chemical constituents in herbs of Anoectochilus roxburghii II].

OBJECTIVE: To study the chemical constituents of Anoectochilus roxburghii. METHOD: Silica gel and sephadex LH-20 column chromotography were used in the isolation from the ethanol extracts of the whole plant, the compounds were determined on the basis of various modern spectroscopic analysis and physical constants. RESULT: Five compounds were isolated from the CHCl3 soluble portion, identified as p-Hydroxybenzaldehyde (I), ferulic acid (II), quercetin (III), daucosterol (IV), cirsilineol (V). CONCLUSION: All these compounds were isolated from the plant for the first time, compound V was isolated from the Orchid Family for the first time, the other compounds were isolated from this genus for the first time.