Sinomenine ameliorates fibroblast-like synoviocytes dysfunction by promoting phosphorylation and nuclear translocation of CRMP2.

ETHNOPHARMACOLOGICAL RELEVANCE: Rheumatoid arthritis (RA) is a chronic autoimmune disease characterized by synovial inflammation and arthritic pain. Sinomenine (SIN), derived from the rhizome of Chinese medical herb Qing Teng (scientific name: Sinomenium acutum (Thunb.) Rehd. Et Wils), has a longstanding use in Chinese traditional medicine for treating rheumatoid arthritis. It has been shown to possess anti-inflammatory, analgesic, and immunosuppressive effects with minimal side-effects clinically. However, the mechanisms governing its effects in treatment of joint pathology, especially on fibroblast-like synoviocytes (FLSs) dysfunction, and arthritic pain remains unclear. AIM: This study aimed to investigate the effect and underlying mechanism of SIN on arthritic joint inflammation and joint FLSs dysfunctions. MATERIALS AND METHODS: Collagen-induced arthritis (CIA) was induced in rats and the therapeutic effects of SIN on joint pathology were evaluated histopathologically. Next, we conducted a series of experiments using LPS-induced FLSs, which were divided into five groups (Naïve, LPS, SIN 10, 20, 50 µg/ml). The expression of inflammatory factors was measured by qPCR and ELISA. The invasive ability of cells was detected by modified Transwell assay and qPCR. Transwell migration and cell scratch assays were used to assess the migration ability of cells. The distribution and content of relevant proteins were observed by immunofluorescence and laser confocal microscopy, as well as Western Blot and qPCR. FLSs were transfected with plasmids (CRMP2 T514A/D) to directly modulate the post-translational modification of CRMP2 protein and downstream effects on FLSs function was monitored. RESULTS: SIN alleviated joint inflammation in rats with CIA, as evidenced by improvement of synovial hyperplasia, inflammatory cell infiltration and cartilage damage, as well as inhibition of pro-inflammatory cytokines release from FLSs induced by LPS. In vitro studies revealed a concentration-dependent suppression of SIN on the invasion and migration of FLSs induced by LPS. In addition, SIN downregulated the expression of cellular CRMP2 that was induced by LPS in FLSs, but increased its phosphorylation at residue T514. Moreover, regulation of pCRMP2 T514 by plasmids transfection (CRMP2 T514A/D) significantly influenced the migration and invasion of FLSs. Finally, SIN promoted nuclear translocation of pCRMP2 T514 in FLSs. CONCLUSIONS: SIN may exert its anti-inflammatory and analgesic effects by modulating CRMP2 T514 phosphorylation and its nuclear translocation of FLSs, inhibiting pro-inflammatory cytokine release, and suppressing abnormal invasion and migration. Phosphorylation of CRMP2 at the T514 site in FLSs may present a new therapeutic target for treating inflammatory joint's destruction and arthritic pain in RA.

[Chemical constituents from Pulsatillae Radix].

Chemical constituents of ethanol extract of Pulsatillae Radix were investigated. The n-butanol fraction of ethanol extract of Pulsatillae Radix was isolated and purified by macroporous resin and silica gel column chromatography and semi-preparative high performance liquid chromatography. The triterpenoid glycosides were identified by multiple spectral methods. Six compounds were obtained from the n-butanol fraction of ethanol extract of Pulsatillae Radix and identified as 23-aldehyde-cussosaponin C(1), cussosaponin C(2), anemoside B4(3), akebia saponin D(4), pulchinenoside E3(5), and hederacoside C(6). Among them, compound 1 was a new compound.

Ultrasonic-assisted extraction, fatty acids identification of the seeds oil and isolation of chemical constituent from oil residue of Belamcanda chinensis.

Belamcanda chinensis is a common garden herb. The extraction technology of B. chinensis seed oil (BSO) was optimized by ultrasonic-assisted extraction (UAE) method, the composition, relative content of main fatty acids and physicochemical properties of BSO were determined, and the isolation, identification and determination of chemical constituent in BSO residue (BSOR) were also investigated. The optimum process conditions of BSO by UAE were optimized as ultrasound time 14 min, extraction temperature 42℃, the ultrasound power 413 W and the liquid-solid ratio 27:1 mL/g. Under this condition, the extraction yield was 22.32 % with the high contents of linoleic acid and oleic acid in BSO. Ten compounds were isolated and identified from BSOR, and belamcandaoid P (9) was a new compound. The contents of the determined compounds were all at high level in B. chinensis seed. The study provided a certain scientific reference for the comprehensive development and utilization of B. chinensis seeds.

Human supplementation with Pediococcus acidilactici GR-1 decreases heavy metals levels through modifying the gut microbiota and metabolome.

Exposure to heavy metals (HMs) is a threat to human health. Although probiotics can detoxify HMs in animals, their effectiveness and mechanism of action in humans have not been studied well. Therefore, we conducted this randomized, double-blind, controlled trial on 152 occupational workers from the metal industry, an at-risk human population, to explore the effectiveness of probiotic yogurt in reducing HM levels. Participants were randomly assigned to two groups: one consumed probiotic yogurt containing the HM-resistant strain Pediococcus acidilactici GR-1 and the other consumed conventional yogurt for 12 weeks. Analysis of metal contents in the blood revealed that the consumption of probiotic yogurt resulted in a higher and faster decrease in copper (34.45%) and nickel (38.34%) levels in the blood than the consumption of conventional yogurt (16.41% and 27.57%, respectively). Metagenomic and metabolomic studies identified a close correlation between gut microbiota (GM) and host metabolism. Significantly enriched members of Blautia and Bifidobacterium correlated positively with the antioxidant capacities of GM and host. Further murine experiments confirmed the essential role of GM and protective effect of GR-1 on the antioxidative role of the intestine against copper. Thus, the use of probiotic yogurt may be an effective and affordable approach for combating toxic metal exposure through the protection of indigenous GM in humans.ClinicalTrials.gov identifier: ChiCTR2100053222.

Could peony seeds oil become a high-quality edible vegetable oil? The nutritional and phytochemistry profiles, extraction, health benefits, safety and value-added-products.

Edible oil provides essential nutrients for human body and plays a very important role in human health. Looking for edible oil resource plants with high content and high-quality oil plays an important role in ensuring the adequate supply of edible oil. Peony seeds oil (PSO) was approved as a new resource food and became an edible vegetable oil for its rich content of fatty acids, total unsaturated fatty acids, linoleic acid and linolenic acid. In addition, PSO also contains high contents of squalene, tocopherol, phytosterol and plant polyphenol. PSO has been reported to present various health benefits including antioxidation, blood lipid reduction, hepatoprotection, immunity regulation, blood sugar control, neuroprotection, anti-inflammatory, cytotoxic activity, therapeutic effects on scald, uvioresistant effects and so on. Toxicity studies showed PSO had great safety without any toxic and side effects, and could be processed to produce PSO microcapsule, microemulsion, nanoemulsion gel, biscuit, daily chemical products, soft capsule and other high value-added-products. Therefore, the present article aims to summarize the research findings regarding to the nutritional and phytochemistry profiles, the extraction methods, health benefits, safety and the high value-added-products of PSO. These informations laid a good foundation for the in-depth research and development of PSO, and also proved that PSO was a high-quality edible vegetable oil with health value. With the deepening of research, the studies on PSO will be more and more comprehensive. It believes that PSO will become an important functional edible oil, and will be more widely used as an important functional food and linolenic acid supplement.

Improving selenium accumulation in broilers using Escherichia coli Nissle 1917 with surface-displayed selenite reductase SerV01.

Selenium levels have a critical impact on livestock and poultry, and selenium nanoparticles (SeNPs) have shown significant efficiency in supplementation. This study identified a high-efficiency selenite reductase, SerV01, in Staphylococcus aureus LZ-01, which can convert Se2O32- to SeNPs. Subsequently, SerV01 was introduced into the intestines of the broilers using the surface display-engineered E. coli Nissle 1917 (EcN). The results showed that the engineered bacteria (EcN-IS) significantly increased the selenium content by 0.87 mg kg-1, 0.52 mg kg-1, and 6.10 mg L-1 in the liver, breast muscle, and serum, respectively. With SeNPs + EcN-IS treatment, glutathione peroxidase and thioredoxin reductase levels reached 0.7536 ± 0.03176 U µL-1 protein and 2.463 ± 0.1685 U µL-1 protein, respectively. With the modified probiotics, the proportion of beneficial intestinal flora increased, with Lactobacillus and Propionibacterium accounting for 75.85% and 0.19%. This technology provides a novel idea to facilitate the exploitation of selenium in broiler diets and improve antioxidant capability.

Comprehensive metabolite profile of multi-bioactive extract from tree peony (Paeonia ostii and Paeonia rockii) fruits based on MS/MS molecular networking.

Tree peony seed, traditionally used for edible oil production, is rich in α-linolenic acid. However, little attention is given to the fruit by-products during seed oil production. The present work aimed to comprehensively investigate the phytochemical constituents and multiple biological activities of different parts of tree peony fruits harvested from Paeonia ostii and Paeonia rockii. 130 metabolites were rapidly identified through UPLC-Triple-TOF-MS on the basis of MS/MS molecular networking. Metabolite quantification was performed through the targeted approach of HPLC-ESI-QQQ-MS. Eight chemical markers were screened via principal component analysis (PCA) for distinguishing species and tissues. Interestingly, two dominant compounds, paeoniflorin and trans-resveratrol, are specially localized in seed kernel and seed coat, respectively. Unexpectedly, the extracts of fruit pod and seed coat showed significantly stronger antioxidant, antibacterial, and anti-neuroinflammatory activities than seed kernel from both P. ostii and P. rockii. Our work demonstrated that tree peony fruit is promising natural source of bioactive components and provided its potential utilization in food and pharmaceutical industries.

[Adjuvant treatment of penetrating moxibustion at governor vessel for persistent allergic rhinitis of deficiency-cold syndrome].

OBJECTIVE: To explore the efficacy and action mechanism of penetrating moxibustion at governor vessel for persistent allergic rhinitis of deficiency-cold syndrome. METHODS: Ninety patients with persistent allergic rhinitis of deficiency-cold syndrome were randomly divided into an observation group (n = 45) and a control group (n = 45). The patients in the control group were treated with momethasone furoate nasal spray, 2 sprays per side per time, once a day. On the basis of treatment in the control group, the patients in the observation group were additionally treated with penetrating moxibustion at governor vessel, 2 h per treatment, once a week. Both groups were treated for 4 weeks. The TCM symptom score, visual analogue scale (VAS) score and rhinoconjunctivitis quality of life questionnaire (RQLQ) score were observed in the two groups before and after treatment. The serum level of immunoglobulin E (IgE) and complete blood count of eosinophil (EOS) were measured before and after treatment, and the clinical effects were compared. RESULTS: Compared before treatment, the TCM symptom scores, VAS scores, RQLQ scores, serum levels of IgE and complete blood count of EOS in the two groups were all reduced after treatment (P<0.05), and those in the observation group were lower than those in the control group (P<0.05). The total effective rate in the observation group was 95.6% (43/45), which was higher than 82.2% (37/45) in the control group (P<0.05). CONCLUSION: Based on the momethasone furoate nasal spray, the adjuvant treatment of penetrating moxibustion at governor vessel could significantly improve the clinical symptoms in patients with persistent allergic rhinitis of deficiency-cold syndrome, and its mechanism may be related to the regulation of immune disorder.

Network pharmacology study on the mechanism of the herb pair of prepared Rehmannia root-Chinese arborvitae kernel for anxiety disorders.

BACKGROUND: Although anxiety disorders are one of the most common mental illness in population, antianxiety drugs often only have single action targets, require long-term use, and are associated with many adverse reactions and dependencies. Professor Yan Zhaojun from Shandong Provincial Hospital of Traditional Chinese Medicine (TCM) has applied the modified Renshu Powder, a TCM formula, to treat anxiety disorders, with satisfactory outcomes. Here, we investigated the mechanism of action of two core herbs (prepared Rehmannia root and Chinese arborvitae kernel) in the Renshu Powder in the treatment of anxiety disorders by using network pharmacology approaches. METHODS: Candidate compounds of the herb pair of prepared Rehmannia root-Chinese arborvitae kernel were extracted via the Traditional Chinese Medicine Systems Pharmacology (TCMSP) platform. The targets of action of the main compounds were collected using the SwissTargetPrediction database. Targets associated with anxiety disorders were retrieved from DisGeNET, Online Mendelian Inheritance in Man (OMIM), DrugBank, GeneCards, and Comparative Toxicogenomics Database (CTD) databases. The compound-target interaction network was constructed by Cytoscape 3.7.2 software, and the protein-protein interaction (PPI) network was constructed using the Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) platform. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses the data by using Metascape. RESULTS: The main active compounds of the herb pair included arachidonic acid, stigmasterol, and beta-sitosterol. The key targets included Nitric Oxide Synthase 3 (NOS3), Epidermal growth factor (EGF), Prostaglandin-Endoperoxide Synthase 2 (PTGS2), Caspase 3 (CASP3), Mitogen-Activated Protein Kinase 1 (MAPK1), Peroxisome proliferator-activated receptor gamma (PPARG), RELA Proto-Oncogene, NF-KB Subunit (RELA), Estrogen Receptor 1 (ESR1), Solute Carrier Family 6 Member 4 (SLC6A4), and Phosphatase and Tensin homolog deleted on chromosome 10 (PTEN). Anxiety disorder-related GO analysis mainly involved synaptic signaling, neurotransmitter receptor activity, and G protein-coupled neurotransmitter receptor activity. The KEGG pathways involved neuroactive ligand-receptor interaction, serotonergic synapse, PI3K/AKT/mTOR signaling pathway, and MAPK signaling pathway. CONCLUSIONS: The mechanism of action of the prepared Rehmannia root-Chinese arborvitae kernel in treating anxiety disorders involves multiple ingredients, multiple targets, and pathways.

Feed-additive of bioengineering strain with surface-displayed laccase degrades sulfadiazine in broiler manure and maintains intestinal flora structure.

Sulfonamide antibiotics (SAs) are excreted into the ecosystem unchanged through feces and urine because of their low adsorption and degradation in the guts of humans and animals. In this study, a novel whole-cell biocatalyst with fungal laccase on the cell surface of Escherichia coli Nissle 1917 was developed to degrade sulfadiazine (SDZ). Engineered strain EcN-IL showed laccase enzyme activity of 2 ± 1 U/mg dry weight of cell and degraded 37 ± 1% of SDZ at temperature 40 °C and pH 5 within 3 h in vitro. Strain EcN-IL with 500 mg/kg of SDZ was employed as a food supplement to feed chicken broilers, which can reduce the residue of SDZ in broiler manure by 58 ± 2% and also reduced dysbiosis of the gut microbiota due to overuse of antibiotics. The genetically engineered EcN-IL has laid a foundation for degrading SDZ in broilers and their manure.

Disturbed mitochondrial dynamics in CD8+ TILs reinforce T cell exhaustion.

The metabolic challenges present in tumors attenuate the metabolic fitness and antitumor activity of tumor-infiltrating T lymphocytes (TILs). However, it remains unclear whether persistent metabolic insufficiency can imprint permanent T cell dysfunction. We found that TILs accumulated depolarized mitochondria as a result of decreased mitophagy activity and displayed functional, transcriptomic and epigenetic characteristics of terminally exhausted T cells. Mechanistically, reduced mitochondrial fitness in TILs was induced by the coordination of T cell receptor stimulation, microenvironmental stressors and PD-1 signaling. Enforced accumulation of depolarized mitochondria with pharmacological inhibitors induced epigenetic reprogramming toward terminal exhaustion, indicating that mitochondrial deregulation caused T cell exhaustion. Furthermore, supplementation with nicotinamide riboside enhanced T cell mitochondrial fitness and improved responsiveness to anti-PD-1 treatment. Together, our results reveal insights into how mitochondrial dynamics and quality orchestrate T cell antitumor responses and commitment to the exhaustion program.

Jiawei Yanghe decoction ameliorates cartilage degradation in vitro and vivo via Wnt/ß-catenin signaling pathway.

Jiawei Yanghe decoction (JWYHD) is a Traditional Chinese Medicine (TCM) formula for the treatment of osteoarthritis (OA), however the underlying mechanisms of action of JWYHD in OA are not fully explored. This study investigates how JWYHD protects cartilage from degradation via Wnt/ß-catenin signaling pathway. The chondroprotective and anti-inflammatory effect of JWYHD on chondrocytes in vitro and on MIA-induced OA rat model in vivo were investigated. In vitro, JWYHD increased the chondrocyte viability against interleukin (IL)-1ß-induced chondrocytes apoptosis and preserved glycosaminoglycans in the extracellular matrix. JWYHD promoted chondrocyte viability against apoptosis, decreased MMP-3, MMP-13, Caspase-3, Caspase-9 via Wnt/ß-catenin signaling pathway in both IL-1ß-induced and Licl-induced chondrocytes. The qRT-PCR and western blot results showed that mRNA and protein expressions of Wnt signaling pathway related genes ß-catenin and CyclinD1, apoptosis related genes Casapase-3 and Caspase-9, collagen degradation related genes Metalloproteinase (MMP)-3 and MMP-13 were up-regulated, and Col2a1 was down-regulated on IL-1ß-induced chondrocytes. Treatment with JWYHD reversed these effects in a dose-dependent manner. Licl was used as Wnt/ß-catenin signaling pathway activator in chondrocytes to determine the molecular mechanisms. Activation of Wnt signaling pathway by Licl up-regulated ß-catenin, CyclinD1, Axin2, Caspase-3, Caspase-9, MMP-3, MMP-13 and IL-1ß. These effects were blocked by JWYHD treatment. Furthermore, 75 Sprawl-Dawley rats were used to verify the results obtained in vitro. A total of 75 rats were randomly divided into the control group (no MIA-induced OA, received intragastric administration of an equivalent amount of saline), the OA group (MIA-induced OA, received intragastric administration of an equivalent amount of saline), and the JWYHD treatment group (MIA-induced OA, received intragastric administration of an equivalent amount of various concentrations of JWYHD at 1.4/2.7/5.5 g/kg). After 8 weeks of administration, all rats were sacrificed. JWYHD decreased the MIA-induced up-regulation of ß-catenin, CyclinD1, Caspase-3, Caspase-9, MMP-3 and MMP-13 protein expressions in cartilage. It was also demonstrated that JWYHD decreased serum and synovium pro-inflammatory cytokines, IL-1ß, IL-6 and TNF-α in MIA-induced OA rats and ameliorated the cartilage degradation. Histopathological staining, macroscopic observation and micro-CT scan with 3-dimension remodeling showed a cartilage protective effect of JWYHD. In conclusion, JWYHD possess multiple capabilities including preventing chondrocyte apoptosis, preserving integrity of extracellular matrix and anti-inflammatory effect in the treatment of OA both in vitro and in vivo.

Profiling and simultaneous quantitative determination of oligostilbenes in Paeonia ostii seed shell from different geographical areas in China and their comparative evaluation.

INTRODUCTION: The Paeonia ostii T. Hong & J. X. Zhang seed shell, characterised by a high content of oligostilbenes, is one of the two most important by-products in the preparation of seed oil. Oligostilbenes are considered characteristic constituents of the genus Paeonia, and can be used in fingerprinting to determine the geographical origin and the quality of raw materials. OBJECTIVE: To develop and optimise a simple and reproducible high-performance liquid chromatography diode array detection (HPLC-DAD) method for the simultaneous determination of seven oligostilbenes in P. ostii seed shell from different geographical areas, and to associate the cultivation area. METHODOLOGY: A validated HPLC method coupled with a DAD detector was performed for the detection and determination of target compounds in the samples. Optimal chromatographic conditions were achieved using an Agilent Zorbax Eclipse SB-AQ-C18 column and a gradient elution with acetonitrile and potassium dihydrogen phosphate solution. RESULTS: The proposed quantitative method showed appropriate accuracy and precision, and was successfully applied to the routine analysis of seven oligostilbenes and the quality evaluation of 50 P. ostii seed shell samples. There were significant differences between the contents of the seven oligostilbenes in different samples (P < 0.01). CONCLUSION: The results demonstrated that the oligostilbenes were main secondary metabolites in the P. ostii seed shells, and the content of seven components in P. ostii seed shells sourced from different cultivation areas in China was different.

Using nano-attapulgite clay compounded hydrophilic urethane foams (AT/HUFs) as biofilm support enhances oil-refinery wastewater treatment in a biofilm membrane bioreactor.

Petroleum refinery wastewater (PRW) treatments based on biofilm membrane bioreactor (BF-MBR) technology is an ideal approach and biofilm supporting material is a critical factor. In this study, BF-MBR with nano-attapulgite clay compounded hydrophilic urethane foams (AT/HUFs) as a biofilm support was used to treat PRW with a hydraulic retention time of 5 h. The removal rate of 500 mg/L chemical oxygen demand (COD), 15 mg/L NH4+ and 180 NTU of turbidity were 99.73%, 97.48% and 99.99%, which were 23%, 20%, and 6% higher than in the control bioreactor, respectively. These results were comparatively higher than that observed for the sequencing batch reactor (SBR). The death rate of the Spirodela polyrrhiza (L.) irrigated with BF-MBR-treated water was 4.44%, which was similar to that of the plants irrigated with tap water (3.33%) and SBR-treated water (5.56%), but significantly lower than that irrigated with raw water (84.44%). The counts demonstrated by qPCR for total bacteria, denitrifiers, nitrite oxidizing bacteria, ammonia oxidizing bacteria, and ammonia-oxidizing archaea were also higher in BF-MBR than those obtained by SBR. Moreover, the results of 16 s rRNA sequencing have demonstrated that the wastewater remediation microbes were enriched in AT/HUFs, e.g., Acidovorax can degrade polycyclic aromatic hydrocarbons, and Sulfuritalea is an efficient nitrite degrader. In summary, BF-MBR using AT/HUF as a biofilm support improves microbiome of the actived sludge and is reliable for oil-refinery wastewater treatment.

Optimization of Ultrasound-Assisted Extraction of Monoterpene Glycoside from Oil Peony Seed Cake.

Ultrasound-assisted extraction (UAE) of total monoterpene glycosides extract (TMGE) from oil peony seed cakes was investigated. The extraction yield was optimized using response surface methodology (RSM). The chemical constituents of the monoterpene glycosides extract were isolated by repeated column chromatography, and the contents of the main isolated monoterpene glycosides in the oil peony seed cakes were determined by HPLC. The optimum conditions were as follows: a liquid-to-solid ratio of 27 mL/g, ultrasonic extraction time of 16 min, ultrasonic extraction temperature of 26 °C, and ethanol concentration of 67%. Under these conditions, the extraction yield of TMGE was 10.24%. Twenty monoterpene glycosides were isolated from the oil peony seed cakes, and compounds 11-12, 16 and 20 showed strong inhibitory activities on NO production. TMGE from oil peony seed cakes can also to be used as promising immunosuppressive drug due to its high content of monoterpene glycosides and immune-inhibitory activity. PRACTICAL AAPPLICATION: The peony seed oil was authorized as a new food by the Ministry of Health of the People's Republic of China. Peony seed cake is one of the most important by-products in the preparation of peony seed oil, and accounts for approximately 40% of the total mass of the peony seed. Total monoterpene glycosides are the main active ingredient of oil peony seed cake. This research has optimized the extraction conditions of total monoterpene glycoside from seeds cake of Paeonia ostii, which will provide useful reference information for further studies, and offer related industries with helpful guidance in practice.

Co-expression of YieF and PhoN in Deinococcus radiodurans R1 improves uranium bioprecipitation by reducing chromium interference.

Overexpression of the enzyme phosphatase (PhoN/PhoK) in the radiation-resistant bacterium Deinococcus radiodurans could be an efficient strategy for uranium remediation. However, the presence of other metals in nuclear wastes often interferes with uranium bioprecipitation. In our study, the uranium-precipitating ability of the PhoN-expressing D. radiodurans strain (Deino-phoN) significantly decreased by 45.4% in 13 h in the presence of chromium (VI); however, it was partially recovered after supplementation with chromium (III). Therefore, the reduction of chromium (VI) to chromium (III) was obtained by the co-expression of the YieF protein and PhoN in D. radiodurans (Deino-phoN-yieF). As a result, an increase in the chromium (VI) reduction (25.1%) rate was observed in 24 h. Furthermore, uranium precipitation also increased by 28.0%. For the decontamination of groundwater, we immobilized Deino-phoN-yieF cells using Polyvinyl alcohol (PVA)-sodium alginate (SA) beads, followed by incubation in a bioreactor. Approximately 99% of chromium (VI) and uranium (VI) was removed after 4 continuous cycles operated for a period of over 20 days at room temperature (25 °C). Therefore, Deino-phoN-yieF could be used as a potential biological agent for mixed radioactive nuclear waste remediation.

Copper (II) binding of NAD(P)H- flavin oxidoreductase (NfoR) enhances its Cr (VI)-reducing ability.

Microbes can reduce hexavalent chromium Cr (VI) to the less toxic and soluble trivalent Cr (III). Copper stimulates microbial reduction of Cr (VI) by the Bacillus, Ochrobactrum, and Gluconobacter species; however, the mechanism remains unclear. In our study, the rate of Cr (VI) reduction by Staphylococcus aureus LZ-01 was increased by 210 % when supplemented with 60 µM Cu (II). A putative NAD(P)H-flavin oxidoreductase gene (nfoR) was upregulated under Cr (VI) stress. NfoR-knockout mutant displayed impaired reduction of Cr (VI) and Cu (II)-enhanced Cr (VI) reduction by nfoR isogenic mutant was attenuated in the presence of Cu (II). In vitro tests showed an increased V max value of 25.22 µM min-1 mg-1 NfoR in the presence of Cu (II). Together, these results indicate that NfoR is responsible for Cu (II) enhancement. Isothermal titration calorimetry (ITC) assays confirmed the interaction of NfoR with Cu (II) at the dissociation constant of 85.5 µM. Site-directed mutagenesis indicates that His100, His128, and Met165 residues may be important for Cu (II) binding, while Cys163 is necessary for the FMN binding of NfoR. These findings show that Cu (II)-enhanced NfoR belongs to a new branch of Cr (VI) reductases and profoundly influences Cr (VI) reduction.

Quantitative Determination of Anthraquinones and Resveratrol in Polygonum Cillinerve (Nakai) Ohwi by HPLC-PAD.

A simple, sensitive, and validated high-performance liquid chromatography-photodiode array method has been established for the simultaneous determination of anthraquinones and resveratrol in Polygonum Cillinerve (Nakai) Ohwi (Zhushaqi in Chinese). The evaluation was performed using a Sunfire C18 reversed-phase column with 30°C column temperature. The mobile phase was composed of a gradient elution of 0.5% acetic acid (solvent A) and methanol (solvent B) with flow rate of 1.0 mL/min. The detection wavelength was at 254 nm. The method developed and validated is simple, shows good linearity, sensitivity, precision, and recovery, and is applied to analyze anthraquinones and resveratrol in 13 batches of Zhushaqi. The results show that the target compounds of Zhushaqi are significantly different among these samples. Based on results of the statistical analysis, the samples collected from Funiu Mountain were clustered together, and the samples obtained from Bozhou Market were close together. The developed method can be a useful tool in quality control and used to evaluate difference and to identify the geographical area of Zhushaqi, and also to provide technical support for the pharmacological and clinical research of related drugs.

Antioxidant Activities and Chemical Constituents of Flavonoids from the Flower of Paeonia ostii.

Paeonia ostii is a traditional medicinal plant popularly used in China. This study intended to evaluate the antioxidant properties and the chemical components of the flavonoid-rich extracts from the flowers of P. ostii. The results showed that the flavonoid-rich extracts from the flowers of P. ostii had strong scavenging capacities on 2,2'-Azinobis-(3-ethylbenzthiazoline-6-sulphonate) (ABTS), hydroxyls, superoxide anions, and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals in a dose-dependent manner. Five flavonoids, dihydrokaempferol (1), apigenin-7-O-ß-d-glucoside (2), apigenin-7-O-ß-d-neohesperidoside (3), kaempferol-7-O-ß-d-glucopyranoside (4), and kaempferol-3-O-ß-d-glucopyranosyl-7-O-ß-d-glucopyranoside (5), were isolated from the flavonoid-rich extracts of the flowers of P. ostii. High-performance liquid chromatography (HPLC) analysis revealed that compounds 3 and 4 were abundant in the P. ostii flower and in flavonoid-rich extracts. The main components of the flower of P. ostii are flavonoids. The high antioxidant activity of the flavonoid-rich extracts may be attributed to the high content of flavonoids. The five isolated flavonoids were the primary antioxidant ingredients, and may play important roles in the strong antioxidant activities of this flower. Based on the obtained results, the flower of P. ostii could be a potential source of natural antioxidants in food and pharmaceutical applications.