Enzymatic synthesis of Vaccinium blue using vaccinoside as a bifunctional precursor.

Since Tang dynasty in China, the fresh leaves of Vaccinium bracteatum (VBL) have been applied as natural pigment to produce black rice. However, detailed information on its biosynthetic mechanism still remained unclear. Following rice dyeing capacity assay, vaccinoside, one of iridoid glycosides, was identified as the key active compound. Increased methodical research demonstrated vaccinoside as a distinct bifunctional precursor, which could be catalyzed by polyphenol oxidase or ß-glucosidase independently, followed by reaction with 15 amino acids to give blue pigments (VBPs; λmax 581-590 nm) of different hues. Two synthetic pathways of VBPs were proposed, using multiple techniques such as HPLC, HPSEC, UV-Vis spectrum and colorimeter as analysis tools. Black rice was interpreted to be prepared by cooking, using vaccinoside, intrinsic enzymes from fresh VBL and rice protein in combination. These findings promote the understanding of VBP formation mechanisms and provide an efficient method of producing novel Vaccinium blue pigments.

Microwave-assisted enzymatic extraction brings a notably high yield of polysaccharides from mountain Zizania latifolia.

Mountain Zizania latifolia is produced at scale in China, and the edible swollen culm is exported to many countries, but little attention has been paid to its functional components. In this work, microwave-assisted enzymatic extraction (MAEE) is used for the first time to extract polysaccharides from mountain Z. latifolia swollen culm (PMZL). MAEE conditions optimized by Box-Behnken design-response surface methodology were as follows: 2.4% cellulase, microwaving for 6.0 min at 607 W, with a liquid-to-solid ratio of 63:1 ml g-1 . Under these conditions, a notably high yield of 60.43% ± 1.12% for PMZL was achieved, which was significantly higher (p < 0.01) than from plain-grown varieties. PMZL are naturally occurring sulfated polysaccharide-protein complexes containing 8.46% ± 0.18% proteins and 7.86% ± 0.73% sulfates. PMZL comprises mannose, glucosamine, rhamnose, glucose, galactose, and arabinose at molar ratios of 3.80:2.68:1.00:17.41:5.12:2.91, with a weight-average molecular weight of 1569,219 Da and a number-average molecular weight of 364,088 Da. The surface morphology of PMZL is composed of tightly packed oval particles, and this kind of promising polysaccharides preferentially scavenges reactive nitrogen species. PRACTICAL APPLICATION: Due to global warming, the land available for planting vegetables is likely to expand to higher areas, so greater attention should now be paid to mountain-grown vegetables. This study provides an efficient way to obtain novel polysaccharides from mountain Zizania latifolia using microwave-assisted enzymatic extraction with a remarkably high yield of 60.4%. This promising source of natural carbohydrates has potential uses in pharmaceutical, nutraceutical, functional foods, cosmetics, and functional materials industries.

Isolation and characterization of secondary metabolites from the leaves of Sauropus spatulifolius Beille and their potential biological assays.

Eight new compounds (1-8), along with three known related compounds (9-11) were isolated from the leaves of Sauropus spatulifolius Beille. Their structures and configurations were elucidated by means of spectrometric and the modified Mosher's method. Among the new compounds, compounds 1 and 2 were identified as ethyl 3, 6-anhydro-2-deoxy-ß-D-arabino-hexofuranoside (1) and ethyl 3, 6-anhydro-2-deoxy- hexofuranoside (2). Compounds 3-5 were the 2-acetylpyrrole derivatives and identified as 2-(2-acetyl-1H-pyrrol-1-yl)-4-hydroxybutyric acid (3), methyl 4-(2-acetyl-lH-pyrrol- 1-yl) butanoate (4) and 1, 4-bis (2-acetyl-1H-pyrrol-1-yl) butane (5), respectively. Compound 6 was elucidated as 7-megastigmane-3, 8, 9-triol. Compounds 7, 8 were identified as kaempferol-3-O-2-deoxy-ß-D-glucoside (7) and kaempferol-3-O-ß-D- glucopyranosyl-(1-6)-2-deoxy-ß-D-glucoside (8). In addition, the cytotoxic activities of all the compounds were also evaluated, where compounds 3, 5, 7, 9\10 and 11 exhibited the magnificent inhibition activity on lung fibroblast differentiation induced by TGF-ß1with low toxicity against the RLE-6TN cell.

The interaction between gut microbiota and flavonoid extract from Smilax glabra Roxb. and its potent alleviation of fatty liver.

Fatty liver is associated with intestinal microbiota dysbiosis and low-grade chronic inflammation. Herein we report the interaction of the flavonoid extract from Smilax glabra Roxb. (FSGR) with gut microbiota. Then, FSGR's function of modulating microbiota in a rat model of high-fat diet (HFD) induced fatty liver has been explored. These investigations indicated that the main compound in FSGR, such as astilbin and its isomers, could be metabolized to aglycone, while further splitting resulted in some phenolic acid compounds through a redox reaction. The data obtained clearly showed that FSGR not only alleviated the steatosis degree of liver cells and modulated the contents of short chain fatty acids (SCFAs) in the intestinal tract, but also reversed gut dysbiosis induced by HFD as prognosticated by the decreased ratio of Firmicutes/Bacteroidetes (F/B) and altered gene expression. The results demonstrated that FSGR probably could be used as a prebiotic agent to impede gut dysbiosis and fatty liver-related metabolic disorders.

A 90-Day Oral Toxicity Study of the Ethanol Extract from Eupatorium japonicum Thunb and Foeniculum vulgare in Rats.

Eupatorium japonicum Thunb and Foeniculum vulgare are two of the most widely used folk herbs and constituents in many traditional Chinese herbal formulas. Nonetheless, little toxicological and safety information associated with following daily repeated exposure is obtained according to previous research. The present study was performed to assess the toxicity of ethanol extract from Eupatorium japonicum Thunb and Foeniculum vulgare (EFE) in male rats administered by dietary oral gavage at target doses of 0.39, 0.78, and 1.56 g/kg body weight/day for 90 days. There were no significant adverse effects on clinical signs, body weight, food conversion efficiency, and vital hematological indices. However, some hematology and biochemical indices such as WCV, MCH, MCHC, LY, MPV, T-CHO, as well as TG revealed significant changes in Sprague-Dawley rats and organ weights in lung and spleen showed diminished in male rats. Necropsy and histopathology findings suggested that no significant differences in absolute weights were found in all organs except lung and spleen, and no treatment-related alteration was identified in any organs. All results obtained in the present study indicated that the proper use of EFE in traditional medicine at oral dosages up to 1.56 g/kg/day body weight may harbor no prolonged toxicity to rats. However, further studies of EFE are still necessary to assess its oral safety in patients.

[Effect of ultrasound-guided acupoint electrical stimulation on diaphragmatic dysfunction associated with mechanical ventilation].

OBJECTIVE: To observe the effect of ultrasound-guided acupoint electrical stimulation on the patients with diaphragmatic dysfunction associated with mechanical ventilation in ICU. METHODS: Fifty-two patients were randomly divided into an observation group (26 cases, 3 cases dropping) and a control group (26 cases). Conventional treatment was given to all patients. On the basis of conventional treatment, acupoint electrical stimulation therapy was applied at Zhangmen (LR 3), Dabao (SP 21), Pishu (BL 20), Shenshu (BL 23), etc. In the observation group, the treatment was given for 30 min each time, 3 times a day for 7 days. Diaphragm thickening fraction (TFdi) was used as an index to guide the individualized setting of stimulation intensity and judge the effect, and the difference of mechanical ventilation time, ICU time, total hospitalization time, hospital mortality and reintubation rate between the two groups were observed. RESULTS: The mechanical ventilation time in the observation group was shorter than that in the control group (P<0.05). There was no significant difference in the ICU time and total hospitalization time between the two groups (P>0.05). During hospitalization, 2 patients died in the observation group and 3 patients died in the control group, there was no significant difference in hospital mortality (P>0.05). One patient in the observation group was reintubated and 8 patients in the control group (P<0.05). The use of acupoint electrical stimulation was a factor in shortening the mechanical ventilation time and reducing the reintubation events (P<0.05). CONCLUSION: Ultrasound-guided acupoint electrical stimulation can relieve ventilator-induced diaphragmatic dysfunction, reduce ventilator support time and reintubation events.

Coreopsis Tinctoria Modulates Lipid Metabolism by Decreasing Low-Density Lipoprotein and Improving Gut Microbiota.

BACKGROUND/AIMS: The prevalence of hyperlipidemia is increasing rapidly. The role of Coreopsis tinctoria (CT) in amending lipid metabolism in hyperlipidemia patients has not been reported. This study aims to evaluate the role of CT in altering lipid metabolism in hyperlipidemia patients and to explore the possible mechanisms mediated by gut microbiota in hyperlipidemia mice models. METHODS: A retrospective analysis in 40 hyperlipidemia patients was conducted, in which 20 patients took fenofibrate and another 20 patients normatively drank water with CT. Hyperlipidemia mice models were also established. Blood biochemical tests were performed using an automatic biochemical analyzer. Liver histopathology was observed by hematoxylin and eosin staining. Ileocecal samples were collected from mice, and bacterial DNA was extracted and sequenced by MiSeq sequencing. Bacterial composition and differences were analyzed. RESULTS: In hyperlipidemia patients, CT was associated with decreased triglyceride and low-density lipoprotein (LDL) levels without liver injury. The experimental hyperlipidemia model also verified a similar result. Gut microbial richness and diversity were significantly decreased in hyperlipidemic mice, but increased after CT treatment. Bacterial communities were significantly differentiated between normal controls and hyperlipidemic mice. CT administration improved gut microbiota composition to an approximately normal status. Meanwhile, CT administration attenuated bacterial alterations at the class, order, family, and genus levels in hyperlipidemic mice. Importantly, the genera Barnesiella, Lactobacillus, and Helicobacter achieved high discriminatory power in hyperlipidemic mice relative to normal controls. CONCLUSIONS: CT can modulate blood lipid metabolism with improvement of liver function by decreasing LDL and improving gut microbiota compositions. These findings may provide novel therapeutic strategies for patients with hyperlipidemia.

Chemical characterization, anti-benign prostatic hyperplasia effect and subchronic toxicity study of total flavonoid extract of Pteris multifida.

The decoction of Pteris multifida had been applied to attenuate symptoms of benign prostatic hyperplasia in Chinese folk medicine. In this study, the total flavonoid extract of Pteris multifida was processed at first. High performance liquor chromatography and tandem mass spectrometer assay revealed 10 flavonoids as key constituents of this extract. After 60-day administration, the total flavonoid extract (180 mg/kg, i. g.) decreased the prostate index in mice of benign prostatic hyperplasia apparently. Immunohistochemical assay revealed inhibition of vascular endothelial growth factor expression, together with activation of transforming growth factor-beta 1 expression in the prostatic samples after administration of the extract. A 90-day subchronic toxicity test was further undertaken in male Sprague-Dawley rats, and the no-observed-adverse-effect level for the extract was 200 mg/kg body weight/day. These results revealed that the total flavonoid extract of Pteris multifida exhibited positive effect with safety, which might be applied in treatment of benign prostatic hyperplasia.

Development and validation of an LC-MS/MS method for simultaneous quantification of levodopa and MD01 in rat plasma and its application to a pharmacokinetic study of mucuna pruriens extract.

Mucuna pruriens, an ancient Indian herbal medicine containing levodopa, is widely used for Parkinson's disease. In order to simultaneously determine levodopa and 1,1-dimethyl-3-carboxy-6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline (MD01) in rat plasma, an improved LC-MS/MS method was developed and validated for a pharmacokinetic study in rats orally administered levodopa or Mucuna pruriens extract (MPE). Elimination of matrix effect and improvement of extraction recovery were achieved through systematic optimization of reversed-phase and hydrophilic interaction chromatographic conditions together with sample clean-up procedures. A satisfactory chromatographic performance was obtained with a Thermo Aquasil C18 column (50 × 2.1 mm, 3 µm) using acetonitrile and water containing 0.2% formic acid as mobile phases. Futhermore, sodium metabisulfite and formic acid were used as stabilizers in neat solutions as well as rat plasma. The method was validated in a dynamic range of 20.0-10,000 ng/mL for levodopa and MD01; the intra- and inter-day precision and accuracy were acceptable. The method was successfully utilized to determine the levodopa level in plasma samples of rats administered levodopa or MPE. Pharmacokinetic results showed that an increase in the AUC of levodopa was observed in rats following oral administration of multiple doses of MPE. Copyright © 2016 John Wiley & Sons, Ltd.

Flavonoids furom Coreopsis tinctoria adjust lipid metabolism in hyperlipidemia animals by down-regulating adipose differentiation-related protein.

BACKGROUND: To identify the chemical structure of Coreopsis tinctoria extracts and their effect and mechanism on reducing blood lipid in hyperlipemia mice. METHODS: The flavonoids were extracted from Coreopsis tinctoria. The chemical structure was identified by HPLC. 59 mice were divided randomly into 5 groups. (group 1: normal diet control; group 2: hyperlipemia model; group 3: hyperlipemia mice treated with Coreopsis tinctoria, low dose 100 mg/kg; group 4: hyperlipemia mice treated with Coreopsis tinctoria high dose group 200 mg/kg; group 5 hyperlipemia mice treated with Fenofibrate. After 2 week of hyperlipid diet, the treatment of Coreopsis tinctoria and Fenofibrate were given for another 6 weeks with continuous hyperlipid diet. The TC, TG, HDL, histology, adipose differentiation-related protein (ADRP) expression in different groups were compared. RESULTS: Compared with normal diet group, TC, TG in hyperlipemia model group increased ( P<0.01). After treatment with Coreopsis tinctoria low dose group, high dose group, TC of the hyperlipemia mice decreased (P<0.05) without increasing AST, ALT and ALP. Fenofibrate can also decrease TC and TG but increase AST, ALT and ALP. Expression of hepatic ADRP increased in hyperlipemia mice. Coreopsis tinctoria high dose group 200 mg/kg can inhibit ADRP as Fenofibrate does. CONCLUSION: The flavonoids from Coreopsis tinctoria extracts can reduce blood lipid without liver function damage, showing better anti- hyperlipemia effect than Fenofibrate by down-regulating ADRP.

Chrysanthemum morifolium extract attenuates high-fat milk-induced fatty liver through peroxisome proliferator-activated receptor α-mediated mechanism in mice.

Some polyphenols derived from plants may ameliorate hyperlipidemic fatty livers; therefore, we hypothesized that polyphenol-rich Chrysanthemum morifolium extract (CME) may exert an inhibitory effect on the formation of hyperlipidemic fatty livers in mice. This study aimed to examine the effects of CME on lipids in blood and liver and on peroxisome proliferator-activated receptor (PPAR)α-mediated gene expression. Mice with hyperlipidemic fatty livers induced by orally administering high-fat milk via gavage and being simultaneously treated with 75 to 300 mg/kg CME for 6 weeks. After CME addition, the serum total cholesterol levels and hepatic weight coefficients decreased, but no significant reduction in the serum triacylglycerol levels were observed. It is important to note that CME might decrease hepatic lipid accumulation, sterol regulatory element binding protein-1c, and fatty acid synthase expression and increase hepatic PPARα, lipoprotein lipase, and cholesterol 7α-hydroxylase expression. However, the expected reduction in hepatic diacylglycerol acyltransferase mRNA expression was not observed. These findings demonstrate that polyphenol-rich CME may prevent hyperlipidemic fatty liver in mice, and its mechanisms may be related to the modulation of sterol regulatory element binding protein-1c, FAS, lipoprotein lipase, and cholesterol 7α-hydroxylase 1 expression through the PPARα-mediated pathway.

Preparative separation of crocins and geniposide simultaneously from gardenia fruits using macroporous resin and reversed-phase chromatography.

Gardenia fruits contain valuable natural food colorants including crocins (gardenia yellow) and geniposide. In this study, a process for the enrichment of crocins and geniposide simultaneously from gardenia fruits was developed using macroporous resin and RP chromatography. The performance of eight different types of macroporous resins was evaluated. Static absorption/desorption experiments revealed that LX60 possessed optimal separating capacity. Further dynamic absorption/desorption experiments on LX60 columns were conducted to obtain the optimal parameters. After one run treatment with LX60, the content of crocin-1 in gardenia yellow reached 29.6%, while geniposide in another fraction reached 83.4%. An extract of crocins was obtained from gardenia yellow in a second-stage separation using RP medium-pressure LC, with its color value to be 756 and the content of crocin-1 reaching 60.8%. The separation process was highly efficient, low cost, and compact, which may be informative for purifications of other natural products from complex plant extracts.

Enrichment and antioxidant properties of flavone C-glycosides from trollflowers using macroporous resin.

Orientin, vitexin and other flavone C-glycosides are functional ingredients abundant in trollflowers. In this study, an efficient method for enrichment of these ingredients from the flowers of Trollius chinensis Bunge was developed using macroporous resin. Separation characteristics of six typical macroporous resins were investigated by static adsorption/desorption and dynamic separation experiments, and HPD450 was selected as optimal one. Dynamic adsorption/desorption experiments on HPD450 columns were conducted to obtain the optimal parameters, followed by a scale-up experiment. Six fractions, TC-1-TC-3 together with their acid hydrolysates were further prepared to evaluate their antioxidant capacities by 2,2-diphenyl-1-picrylhydrazyl radical and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonate) radical cation scavenging assays. They all have notable concentration-dependent antioxidant activities, with TC-1 showing strong antioxidant capacity higher than orientin. The separation process was high-efficient, low-cost and environmental-friendly, which could afford a potential approach for industrial applications.

Flavone C-glycosides from the flowers of Trollius chinensis and their anti-complementary activity.

Phytochemical investigation of ethanol extract from the flowers of Trollius chinensis Bunge resulted in the isolation of two new flavone C-glycosides (1-2), along with 10 known compounds (3-12). The structures of the new compounds were established as 6‴-(3-hydroxy-3-methylglutaroyl)-2″-O-ß-d-galactopyranosyl orientin (1) and 6‴-(3-hydroxy-3-methylglutaroyl)-2″-O-ß-d-galactopyranosyl vitexin (2) on the basis of various spectroscopic analysis (including different 1D and 2D NMR spectroscopies, high-resolution electrospray ionization mass spectrometry) and chemical evidences. Bioassay showed that eight flavonoids inhibited complement activation on the classic pathway in vitro, with their IC50 values ranging from 0.88 to 4.02 mM, which may contribute to the applications of the herb in treatment of acute respiratory distress syndrome, etc.

Preparative purification of peoniflorin and albiflorin from peony rhizome using macroporous resin and medium-pressure liquid chromatography.

Peoniflorin (PF) and albiflorin (AF) are two principal components of Paeonia species, which exhibit various biological activities such as improvement of blood circulation and immunoregulating function. To further utilization of waste parts of peony plants, an efficient method for preparative purification of these two ingredients from white peony rhizome was developed based on macroporous resin (MAR) and medium-pressure liquid chromatography (MPLC). The separation characteristics of nine typical MARs were investigated by static adsorption/desorption experiments, and LX38 was revealed as optimal one. Further static experiments with LX38 resin indicated that the adsorbents fitted well to the pseudo-second-order kinetics model and both Langmuir and Freundlich isotherm models. Based on the optimal process parameters, a large-scale preparation was successfully applied. After one run treatment with LX38, the contents of PF and AF were increased 15-fold to 24.5 and 16.8% in the refined extract, respectively. Both purified compounds were obtained from refined extract by reversed-phase MPLC at second-stage separation. The process developed is better because of its low cost, high efficiency, and procedural simplicity making it a potential approach for large-scale production of PF and AF for their further applications in functional foods and pharmaceuticals.

New triterpenoid saponins from Patrinia scabiosifolia.

Phytochemical investigation of methanol extract from the whole plants of Patrinia scabiosifolia Fisch. resulted in the isolation of three new triterpenoid saponins (1-3) along with twelve known triterpenoids (4-15). The structures of the new compounds were established as 11α, 12α-epoxy-3-O-ß-D-xylopyranosyl-olean-28, 13ß-olide (1), 11α, 12α-epoxy-3-O-ß-D-xylopyranosyl-(1 → 3)-α-L-rhamnopyranosyl-(1 → 2)-ß-D-xylopyranosyl-olean-28, 13ß-olide (2), and 3-O-ß-D-xylopyranosyl-(1 → 3)-α-L-rhamnopyranosyl-(1 → 2)-ß-D-xylopyranosyl oleanolic acid 28-O-ß-D-glucopyranoside (3) on the basis of various spectroscopic analyses (including different 1D and 2D NMR spectroscopies and high-resolution electrospray ionization mass spectrometry) and chemical evidences.

New triterpenoid saponins from Patrinia scabiosaefolia.

Phytochemical investigation of the methanol extract from the whole plants of Patrinia scabiosaefolia Fisch. resulted in the isolation of four new triterpenoid saponins (1-4) along with six known compounds (5-10). On the basis of spectroscopic and chemical methods, the structures of the new compounds were established as 3-O-ß-D-xylopyranosyl-(1→3)-α-L-rhamnopyranosyl-(1→2)-ß-D-xylopyranosyl-12ß,30-dihydroxy-olean-28,13ß-olide (1), 3-O-α-L-rhamnopyranosyl-(1→2)-ß-D-xylopyranosyl-12ß,30-dihydroxy-olean-28,13ß-olide (2), 3-O-ß-D-xylopyranosyl-(1→2)-ß-D-glucopyranosyl-12ß, 30-dihydroxy-olean-28,13ß-olide (3), and 3-O-ß-D-glucopyranosyl-(1→4)-ß-D-xylopyranosyl-(1→3)-α-L-rhamnopyranosyl-(1→2)-ß-D-xylopyranosyl-oleanolic acid 28-O-ß-D-glucopyranoside (4), respectively. Compounds 1-3 possess a novel 12ß,30-dihydroxy-olean-28,13ß-lactone aglycone and a 12ß-hydroxy substituent that is rarely found in this kind of triterpenoid saponin.

A new minor triterpene saponin from kaixin-san prescription.

A new minor oleanane-type saponin, ginsenoside-R(OA) (1), has been isolated from the extract of Kaixin-San, a prescription composed of Panax ginseng and other medicinal herbs, together with 15 known ginsenosides. The structure of 1 was established as 3-O-beta-D-glucopyranosyl-(1-->2)-beta-D-glucuronopyranosyl oleanolic acid 28-O-beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranoside by chemical and spectroscopic evidence. In addition, the presence of ginsenoside-R(OA) in ginseng was confirmed by HPLC-ESI-MS.