[Effect of Antibiotic-Degrading Bacteria on Maturity and Bacterial Community Succession During Pig Manure Composting].

The inoculation of antibiotic-degrading bacteria into manure could promote the removal of antibiotics during composting. However, knowledge on the impact of inoculating these antibiotic-degrading bacteria on the composting process and indigenous microbial community succession is still limited. This study assessed the antibiotic removal efficiency in pig manure after inoculating a microbial inoculum with antibiotic-degrading bacteria as the key component. The effect of inoculating this microbial inoculum on the physicochemical dynamics and the succession of the manure bacterial community during composting was also analyzed. The results showed that the antibiotic degradation in pig manure reached 81.95% after inoculating the microbial inoculum. When compared with that in the control, the total concentration of antibiotic residues in manure with the microbial agent inoculated was decreased by 42.18%. During composting, inoculating the microbial inoculum accelerated the temperature rise of compost, favored water loss, and alleviated the release of NH3 and H2S. Moreover, the total nutrient content (nitrogen, phosphorus, and potassium) in the final compost and the germination index of radish seeds increased by 6.80% and 68.33%, respectively, after inoculating this microbial inoculum. Furthermore, inoculating the microbial inoculum increased the content of stable organic carbon in the final compost and decreased the content of recalcitrant substances such as cellulose and hemicellulose. The analysis of the manure bacterial community showed that inoculating the microbial inoculum increased the relative abundances of Actinomycetes and Firmicutes in the compost. In particular, the thermophilic bacteria that was positively related to the compost temperature was increased significantly (P<0.01) after inoculating the microbial inoculum, whereas the relative abundance of pathogenic bacteria was correspondingly decreased. Network analysis of the bacterial coexistence pattern showed that inoculating this microbial inoculum also changed the interaction pattern of indigenous manure bacterial communities, which greatly reduced the complexity and connectivity of the bacterial interaction and improved the ecological relationship between beneficial bacteria and other bacterial communities. The effect of this microbial inoculum on the interaction with manure bacterial community laid a foundation for the establishment of a new and healthier composting bacterial community. This study provides a scientific basis for the application and development of multifunctional antibiotic-degrading microbial agents in manure treatments.

Trapa natans pericarp extract ameliorates hyperglycemia and hyperlipidemia in type 2 diabetic mice

Abstract The pericarp of Trapa natans L., an annual aquatic floating herb belonging to Lythraceae family, is used as a folk medicine in China. In this study, extracts of Trapa natans pericarp were tested both in vitro and in vivo through a high-fat diet with a single medium dosage streptozotocin injection induced type 2 diabetic mice. Different solvent extracts of Trapa natans pericarp showed α-amylase and α-glucosidase inhibitory activity. After four weeks administration, the ethyl acetate extract of Trapa natans pericarp (50 and 100 mg/kg b.w.) reduced fasting blood glucose level, ameliorated oral glucose tolerance and insulin resistance, improved serum lipids alterations in type 2 diabetic mice as well. Additionally, ethyl acetate extract significantly elevated the insulin receptor substrate 1 and Akt serine/threonine kinase phosphorylation compared to diabetic group. HPLC-MS and HPLC-DAD analysis showed that the ethyl acetate extract was rich in hydrolysable tannins. Results support the notion that Trapa natans pericarp extract has a potential hypoglycemic activity.

Antitussive and expectorant properties of growing and fallen leaves of loquat (Eriobotrya japonica)

ABSTRACT Folium Eriobotryae, the dried leaves of loquat (Eriobotrya japonica, (Thunb.) Lindl., Rosaceae), is a traditional Chinese medicine used to treat cough with phlegm in China. Fallen and growing loquat leaves were tested for their effect on coughing and expectoration in mice. HPLC-ELSD and HPLC-MS analyses of aqueous and ethanol extracts of fallen or growing leaves were used to identify the chemical components responsible for this effect. Both the aqueous and ethanol extracts of growing and fallen leaves of loquat contained antitussive and expectorant activities. Moreover, an aqueous extract of growing loquat leaves with a higher flavonoid content displayed a stronger expectorant activity while the ethanol extract of fallen loquat leaves that contained a higher content of triterpenoid acids induced a stronger antitussive activity.

Detection and Toxicity Evaluation of Pyrrolizidine Alkaloids in Medicinal Plants Gynura bicolor and Gynura divaricata Collected from Different Chinese Locations.

Two edible plants in Southeast Asia, Gynura bicolor and G. divaricata, are not only known to be nutritive but also useful as medicinal herbs. Previous phytochemical investigation of Gynura species showed the presence of hepatotoxic pyrrolizidine alkaloids (PAs), indicating the toxic risk of using these two plants. The present study was designed to analyze the distribution of PA components and tried to evaluate the preliminary toxicity of these two Gynura species. Eight samples of G. bicolor and G. divaricata from five different Chinese locations were collected and their specific PAs were qualitatively characterized by applying an UPLC/MS/MS spectrometry method. Using a pre-column derivatization HPLC method, the total retronecine ester-type PAs in their alkaloids extracts were quantitatively estimated as well. Finally, their genotoxicity was investigated with an effective high-throughput screening method referred to as Vitotox™ test and their potential cytotoxicity was tested on HepG2 cells. It was found that different types of PAs were widely present in Gynura species collected from south of China. Among them, no significant genotoxic effects were detected with serial concentrations through the present in vitro assay. However, the cytotoxicity assay of Gynura plants collected from Jiangsu displayed weak activity at the concentration of 100 mg/ml. It is important to note that this research validates in part the indication that the use of Gynura species requires caution.

Natural Triterpenoids for the Treatment of Diabetes Mellitus: A Review.

Triterpenoids, an important group of secondary metabolites, are widely distributed in nature. Many triterpenoids have been found with potential therapeutic effect against diabetes mellitus. However, the use of triterpenoids for the treatment of diabetes has not been systematically discussed previously. This review summarized the anti-diabetic activity of natural triterpenoids reported since the late 1980s with the emphasis on the molecular mechanisms.

Simultaneous determination of five triterpene acids in rat plasma by liquid chromatography-mass spectrometry and its application in pharmacokinetic study after oral administration of Folium Eriobotryae effective fraction.

Folium Eriobotryae effective fraction (FEA), the extract of Folium Eriobotryae, had been used as anti-hyperglycemia and anti-hyperlipemia medicine in China. A previous study indicated that euscaphic acid, maslinic acid, corosolic acid, oleanolic acid and ursolic acid, the five structurally similar triterpene acids (containing two groups of structural isomers), are the major components of FEA. In the present study, we developed a specific and reliable LC-MS method for simultaneous determination of the five triterpene acids in rat plasma, and further investigated their pharmacokinetic properties after oral administration of FEA. Following a simple sample preparation, chromatographic separation was achieved on a C18 column with a mobile phase composed of methanol-0.1% ammonium acetate (80:20, v/v). Quantification was achieved by monitoring the selected ions at m/z 487.6 for euscaphic acid, m/z 471.5 for maslinic acid and corosolic acid, m/z 455.5 for oleanolic acid and ursolic acid and m/z 469.5 for internal standard. The method was validated to be specific, accurate and precise over the concentration ranges of 10-3000 ng/mL with limits of detections of 5 ng/mL for the five triterpene acids. Finally, the method was successfully applied to the pharmacokinetic study of the five structurally similar triterpene acids in rats after oral administration of FEA.

Natural flavonoids as potential herbal medication for the treatment of diabetes mellitus and its complications.

Diabetes mellitus, together with its various complications, is becoming a serious threat to human health. Natural products are secondary metabolites widely distributed in plants, having a broad range of biological activities. The development of antidiabetic medication from natural products, especially those originating from plants with a traceable folk-usage history in treating diabetes, is receiving more attention. Many studies highlighted not only the benefits of natural flavonoids with hypoglycemic effects, but also their importance in the management of diabetic complications. This review describes selected natural flavonoids that have been validated for their hypoglycemic properties, together with their mechanisms of action. Also discussed are their activities in the treatment of diabetic complications demonstrated via laboratory diabetic animal models, in vitro and clinical trials using human subjects. Published papers from 2000 to date on flavonoids and diabetes were covered through accessing Web of Science and multiple databases for biomedical sciences. The major potential benefits of natural flavonoids discussed in this review clearly suggest that these substances are lead compounds with sufficient structural diversity of great importance in the antidiabetic drug developing process.

Profiling and elucidation of the phenolic compounds in the aerial parts of Gynura bicolor and G. divaricata collected from different Chinese origins.

Gynura bicolor and G. divaricata are not only known to be nutritive as cultured vegetables, but also beneficial as folk medicines in East Asia. As demonstrated by the current phytochemical knowledge, the genus Gynura is a promising source of phenolics with multiple medicinal activities. To expand this phytochemical knowledge, the phenolic secondary metabolites of G. bicolor and G. divaricata were studied. From the aerial parts of these two species, collected in five different Chinese locations, two fractions of phenolic compounds with different polarity were obtained by extraction and chromatographic separation. Using UPLC/MS/MS analysis, a total of 53 phenolics were either identified by comparison with respective reference compounds or tentatively characterized by their chromatographic behavior, UV-absorption patterns, and MS fragmentations. Some naturally existing positional isomers of O-caffeoylquinic acid, O-p-coumaroylquinic acid, O-feruloylquinic acid, and dicaffeoylquinic acid as well as their methyl esters were qualitatively characterized by their specific fragmentation patterns in targeted MS/MS. In addition, the aerial parts of the two Gynura species contained kaempferol, quercetin oligoglycosides, and a variety of derivatives of benzoic acid, hydroxycinnamic acid, and caffeic acid. Furthermore, the distribution of phenolic compounds in the two species from different Chinese origins was discussed. Finally, an investigation of the total phenolic content and in vitro antioxidant activity of the various phenolic fractions was completed, to evaluate the potential of the extracts of these species for medicinal development. The free-radical-scavenging activities of the extracts derived from plants originating from Nanjing were proven to be higher than those of the other extracts, which correlated well with their total phenolic content.

[Response surface optimization of ultrasonic-assisted pigment extraction from Coreopsis tinctoria].

Response surface methodology (RSM) was used to optimizing the ultrasonic-assisted extraction technology of pigment from Coreopsis tinctoria. The results showed that the flavonoids were the main constituents of the pigment Based on single factor experiments, a four-factor-level experiment design were developed by box-benhnhen central composite design method with causal factors of ultrasonic temperature, ultrasonic time, ratio of liquid to raw material, the concentrations of ethanol in solvent and the extract absorbance value for the response. The interactive effects of four crucial technological parameters were assessed by response surface methodology (RSM). The optimum ultrasonic-assisted extraction conditions were as follow: ultrasonic temperature was 70 °C, ultrasonic time was 60 min, the concentrations of ethanol in solvent was 72.25% and the ratio of liquid to raw material was 32.05:1 mL . g-1. Under the optimum extraction technology conditions, the absorbance value was 0. 936. The conditidns are suitable for the extraction process regression analysis and parameter optimization.

Sulfonic acid functionalized nano-γ-Al2O3: a new, efficient, and reusable catalyst for synthesis of 3-substituted-2H-1,4-benzothiazines.

A simple and efficient synthetic protocol has been developed for the synthesis of 3-substituted-2H-1,4-benzothiazines by using a novel sulfonic acid functionalized nano-γ-Al2O3 catalyst, devoid of corrosive acidic, and basic reagents. The developed method has the advantages of good to excellent yields, short reaction times, operational simplicity, and a recyclable catalyst. The catalyst can be prepared by a simple procedure from inexpensive and readily available nano-γ-Al2O3 and has been shown to be recoverable and reusable up to six cycles without any loss of activity.

Investigation of the volatile constituents of different Gynura species from two Chinese origins by SPME/GC-MS.

GC-MS analyses of the volatile constituents obtained by solid phase microextraction (SPME) of two Gynura species, i.e., G. divaricata and G. bicolor, collected from Nanjing and Nanping areas in the east of China, enabled the identification of more than 50 different components. Generally, a higher contents of sesquiterpene compounds such as beta-caryophyllene, alpha-caryophyllene and alpha-copaene were found in G. bicolor than in G. divaricata, regardless of their origin. Qualitatively, the detected major volatiles of G. bicolor and G. divaricata originating from Nanjing were the same, i.e., beta-caryophyllene, alpha-caryophyllene, alpha-copaene, alpha-pinene and beta-pinene. The volatile profile of a third species, G. medica, also originating from Nanjing was different with sesquiterpenes gamma-cadinene, beta-caryophyllene, elixene and monoterpene limonene as the major components. The volatile profiles of G. bicolor and G. divaricata originating from Nanping were more diverse. For both species beta-caryophyllene, alpha-copaene and limonene were the major volatiles, but G. bicolor also produced gamma-caryophyllene and carvone as major constituents, whereas for G. divariata o-cymene was detected. Overall, these results indicate that the biosynthesis of volatiles by Gynura species is not only species related, but is also possibly influenced by the local environmental conditions of growth.

Chemical constituents from the aerial parts of Gynura bicolor.

Gynura bicolor (Willd.) DC., is used in folk recipes for the treatment of diabetes mellitus in Jiangsu, Zhejiang and Sichuan province in the south of China. A previous pharmacological study proved that the plant showed significant hypoglycemic activity on normal and alloxan-diabetic mice. In this study, two terpenes, four megastigmane-type norisoprenoids and two glycosides were isolated from the aqueous ethanolic extract of the aerial parts of Gynura bicolor and characterized mainly by NMR spectroscopy and mass spectrometry. Thes e compounds were isolated for the first time from this plant, and no evidence could be found for the previous reported presence of megastigmane-type norisoprenoids in the genus Gynura.

Two new prenylated isoflavones from Sedum aizoon L.

Investigation of a methanol extract from the Chinese endemic species Sedum aizoon L. led to the isolation of two new isoflavone derivatives that were named sedacin A and B (1 and 2). Their structures were elucidated by the basis of spectroscopic analysis. Two new isoflavone derivatives were isolated compounds were assayed for antioxidant activity.

Studies on the chemical constituents of Anabasis aphylla L.

The investigation on the chemical constituents of Anabasis aphylla L. was carried out by using various chromatographies, such as silica gel, Sephadex LH-20 and RP-C18 column chromatography. Further detailed investigation on the fraction of the ethanol extract of Anabasis aphylla L. yielded one new compound p-acetyl-phenol 1-O-beta-D-xylopyranosyl-(1-->2)-beta-D-glucopyranoside (1), together with five known compounds: piceine (2), isorhamnetin (3), quercetin (4), rutin (5) and isorhamnetin-3-rutinoside (6). Their structures were elucidated by spectral analysis such as NMR and MS. Among these compounds, compounds 2-6 were isolated from this plant for the first time.

The anti-hyperglycemic effect of plants in genus Gynura Cass.

Gynura divaricata (L.) DC and G. bicolor DC are used as secret recipes to treat diabetes mellitus in some parts of China. Pharmacological tests were performed to prove the anti-hyperglycemic effect of these two plants of genus Gynura Cass. in this study. Both water and 95% ethanol extracts of fresh G. divaricata had significant effects on lowering blood glucose level in normal mice, in which the dose of 0.4 g (crude drug)/kg of 95% ethanol extract was more effective than 50 mg/kg glyburide. The ethyl acetate and n-butanol extracts of dried G. divaricata had significant effects on lowering blood glucose level in normal and alloxan diabetic mice too. Both ethyl acetate and n-butanol extracts of dried G. bicolor showed very significant effect on lowering blood glucose level to normal and alloxan-diabetic mice, and the dose 4.0 g (crude drug)/kg had a more hypoglycemic effect than 50 mg/kg glyburide in normal mice.

Euscaphic acid, a new hypoglycemic natural product from Folium Eriobotryae.

Folium Eriobotryae has been used as a medicinal plant for a long time, and it is known to have many physiological actions such as anti-inflammatory, anti-tussive, expectorant and anti-diabetic. We have reported that the 70% ethanol extract of Folium Eriobotryae exerted a significant hypoglycemic effect to alloxan-diabetic mice. In this study, we isolated euscaphic acid, a natural product from Folium Eriobotryae, and investigated its hypoglycemic effect in normoglycemic and alloxan-diabetic mice. All effects had been compared with those of gliclazide. The plasma glucose levels were significantly lowered in normoglycemic mice treated with euscaphic acid compared to mice treated with 0.5% CMC-Na solution only. Moreover, the dosage of 50 mg/kg exerted a significant (P < 0.05) hypoglycemic effect in alloxan-diabetic mice after orally administration. The research proved that euscaphic acid is one of the active hypoglycemic constituents in Folium Eriobotryae, but the details of the mechanism need to be investigated further.

[Studies on the triterpenes from loquat leaf (Eriobotrya japonica)].

OBJECTIVE: To investigate the chemical constituents from Loquat leaf (Eriobotrya japonica). METHODS: Compounds were isolated by a combination of silica gel and ODS column chromatographies and preparative TLC and HPLC, their structures were identified by chemical and spectral methods. RESULTS: Nine triterpenes were isolated and identified as methyl betuliate (I), oleanolic acid (II), ursolic acid (III), methyl maslinate (IV), methyl corosolate, (methyl 2alpha-3alpha-dihydroxyurs-12-en-28-oate, V), maslinic acid (VI), corosolic acid (VII), tormentic acid (VIII), euscaphic acid (IX) from loquat leaf. CONCLUSION: Compounds I and V are obtained from this plant and Eriobotrya genus for the first time.

Pharmacological studies on anti-hyperglycemic effect of folium eriobotryae.

Folium eriobotryae, dried leaves of Eriobotrya japonica (Thunb.) Lindl. is a traditional Chinese medicine with rich resources in China. This research investigated the anti-hyperglycemic effect of folium eriobotryae on normal and alloxan-diabetic mice. The 70% ethanol extract of folium eriobotryae (EJA-0) in doses of 15, 30 and 60 g (crude drug)/kg exerted a significant hypoglycemic effect on alloxan-diabetic mice, among which 30 g/kg of EJA-0 was more effective than 100 mg/kg of phenformin. The total sesquiterpenes (EJA-1) 30 g (crude drug)/kg had significant effect on lowering blood glucose level in normal or/and alloxan-diabetic mice. The tests of maximum dosage and acute toxicity showed that EJA-1 was safe (MD = 360 g/kg, LD(50) = 400.1 g/kg). The pharmacological tests on anti-hyperglycemic effects of EJA-0 and EJA-1 prove that folium eriobotryae is an outstanding material to develop medicine for treatment of diabetes mellitus.