Fabrication of nano-hammer shaped CuO@HApNWs for catalytic degradation of tetracycline.

With widespread and excessive use of antibiotics in medicine, poultry farming, and aquaculture, antibiotic residues have become a significant threat to both eco-environment and human health. In this paper, using hydroxyapatite nanowires (HApNWs) as an ecologically compatible carrier, a novel nano-hammer shaped conjunction with HApNW conjugating CuO microspheres (CuO@HApNWs) was successfully synthesized by in-situ agglomeration method. The catalytic degradation performance of the nano-hammer shaped CuO@HApNWs with Fenton-like activation was investigated by using tetracycline (TC) as a representative antibiotic pollutant. Remarkably, it exhibited excellent catalytic activity, which the removal rate of TC got to 92.0% within 40 min, and the pseudo-second-order reaction kinetic constant was 18.33 × 10-3 L mg-1·min-1, which was 26 times and 5 times than that of HApNWs and CuO, respectively. Furthermore, after recycling 6 times, the degradation efficiency of TC still remained above 85 %. Based on radical scavenger tests and electron paramagnetic resonance (EPR) spectroscopy, it demonstrated that the excellent activity of CuO@HApNWs was mainly attributed to the fact that Fenton-like activation promotes the circulation of Cu2+ and Cu+, the generated main active oxygen species (•OH and O2-•) achieve efficient degradation of TC. In summary, the nano-hammer shaped CuO@HApNWs could be in-situ synthesed, and used as an eco-friendly Fenton-like catalyst for effectively catalytic degradation of organic pollutants, which has great potential for wastewater treatment.

Myeloid-derived suppressor cells ameliorate liver mitochondrial damage to protect against autoimmune hepatitis by releasing small extracellular vesicles.

BACKGROUND: Autoimmune hepatitis (AIH) is an inflammatory liver disease that is associated with impaired self-tolerance. Myeloid-derived supprfessor cells (MDSCs) have been considered to exert counterregulatory effects on AIH. However, the specific mechanism underlying these effects is unclear. Herein, we investigated the efficacy and safety of MDSCs in protecting against AIH and explored the underlying mechanism. METHODS: Circulating and liver MDSC expression levels in 71 AIH patients and 47 healthy control (HC) individuals were detected by flow cytometry and immunohistochemistry. The adoptive transfer of induced bone marrow-derived MDSCs (BM MDSCs) to AIH mice was used to explore the function of MDSCs. Hepatic injury and mitochondrial damage were evaluated by transaminase levels, histopathology, immunohistochemistry, transmission electron microscopy and western blotting. MDSCs were pretreated with the small extracellular vesicle (sEV) generation inhibitor GW4869 to explore the mechanism. Importantly, sEVs derived from MDSCs and MDSCs-GW4869 were injected into model mice to monitor mitochondrial function and biogenesis. RESULTS: Circulating and liver MDSCs were expanded in AIH patients and mouse model. Furthermore, the follow-up data of AIH patients showed that immunosuppressive therapy further promoted the expansion of MDSCs. More importantly, the adoptive transfer of BM MDSCs to AIH mice effectively ameliorated liver injury and regulated the imbalance of the immune microenvironment. Additionally, BM MDSCs reduced liver mitochondrial damage and improved mitochondrial biogenesis. Mechanistically, sEVs derived from BM MDSCs showed the same biological effects as cells, and blocking sEV production weakened the function of BM MDSCs. Finally, multiple long-term administrations of BM MDSCs were proven to be safe in general. CONCLUSION: In conclusion, MDSCs ameliorate liver mitochondrial damage to protect against autoimmune hepatitis by releasing small extracellular vesicles.

Sea cucumber (Acaudina leucoprocta) peptides extended the lifespan and enhanced antioxidant capacity via DAF-16/DAF-2/SOD-3/OLD-1/PEPT-1 in Caenorhabditis elegans.

The sea cucumber peptides (SCPs) from Acaudina leucoprocta were derived from the patented bio-enzyme digestion technology and the molecular weight of obtained SCPs was < 10 kDa. In this study, we investigated the possible anti-aging effects of SCPs on the model of Caenorhabditis elegans and the underlying mechanisms. SCPs extend the average lifespan of nematodes by 31.46%. SCPs enhance the anti-stress capacity of C. elegans by improving heat resistance and mobility, Also, the accumulated potential oxidative stress inducers like lipofuscin and reactive oxygen species (ROS) were reduced to 40.84 and 71.43%. In addition, SCPs can increase the antioxidant capacity in nematodes by enhancing the activity of SOD and CAT and reducing MDA accumulation in nematodes to 32.44%. Mechanistically, SCPs could mediate DAF-16/DAF-2/SOD-3/OLD-1/PEPT-1 axis to improve antioxidant capacity and extend lifespan in nematodes. Taken together, these findings provide a direction for the anti-aging effects of sea cucumber peptides and new insights into the further purifications of SCPs and future research on aging.

Metabolic heterogeneity caused by HLA-DRB1*04:05 and protective effect of inosine on autoimmune hepatitis.

Autoimmune hepatitis (AIH) is an autoimmune disease caused by disruption of liver immune homeostasis. Genetic studies have revealed the predisposition of AIH with the human leukocyte antigen (HLA) region. Recently, metabolomics integrated with genomics has identified many genetic loci of biomedical interest. However, there is no related report in AIH. In the present study, we found that HLA-DRB1*04:05 was linked to the clinical features and prognosis of AIH in Chinese patients. Furthermore, our patients were divided into DRB1*04:05 positive and DRB1*04:05 negative groups and the metabolic profiling was done by HPLC/MS. We chose inosine, one of the highly altered metabolites, to explore the effect on an acute severe hepatitis murine model. The results showed that inosine treatment attenuated hepatocyte apoptosis, enhanced antioxidant ability and inhibited the activation and glycolysis of CD4+ T cell. We propose that inosine participates in the regulation of AIH through its protective effect on hepatocytes and inhibition of overactivated immune cells, which might provide a potential novel approach in treating acute form of AIH.

Walnut Meal Extracts Rich In Polyphenols Mitigate Insulin Resistance and Modulate Gut Microbiota in High Fat Diet-Fed Rats.

Walnut kernel is a traditional Chinese herb recorded in the Chinese Pharmacopoeia with the efficacies of invigorating kidney, tonifying lung, and relaxing bowel. However, the potential mechanisms were unclear. This article aims to uncover the interdict mechanisms of walnut meal extracts (WMP) on high-fat diet (HFD) induced metabolic disorders in rats and reveal how the WMP benefits are associated with changes in the intestinal flora. Sprague-Dawley (SD) rats were fed a standard chow diet or an HFD for 18 weeks. After 6 weeks, the HFD rats were supplemented with 750 mg WMP/kg body weight or the vehicle for 12 weeks. The structure of gut microbiota was assessed by analyzing 16S rDNA sequences. WMP suppressed the weight gain and visceral obesity. WMP treatment also improved lipid profiles and increased antioxidative activities. WMP fully reversed hepatic steatosis with the upregulation of adipocytokines involved in lipid catabolism (e.g., adiponectin, PPAR-γ, visfatin, CEBPα) and the increased activities of lipoprotein lipase and hormone-sensitive lipase, which were associated with glucose tolerance improvement and insulin resistance (IR) mitigation. As revealed by 16S rDNA sequencing, WMP restored the diversity of intestinal flora reduced by HFD. WMP dramatically reduced the abundance of Gram-negative bacteria, especially Fusobacterium varium and Bacteroides vulgatus, and sharply increased the abundance of Lactobacillus animalis decreased by HFD. Our findings demonstrated that WMP suppressed the weight gain and adiposity in HFD-fed rats and fully reversed HFD induced IR and hepatic steatosis while dramatically reducing the abundance of Fusobacteriaceae and Enterobacteriaceae, underscoring the gut-liver axis as a primary target of walnut polyphenols.

Allergenicity of alcohol-soluble wasp pupal proteins and its impact on the gut microbiota.

Wasp pupa protein has triggered allergies in certain consumers. In this study, we investigated the allergenicity of alcohol-soluble wasp pupa protein (ACWP) and its effect on the gut microbiota of mice in vivo. It was found that ACWP caused skin erythema and diarrhea in mice, the up-regulation of HIS, IgE, IL-4, IL-22, and IL-17A, and down-regulation of IgG2a, IgA, and IL-2 in serum. The results also revealed that the thymus, spleen, and small intestine structures in mice also altered significantly, and the intestinal wall structure was disrupted. Fluorescein isothiocyanate (FITC) labeled dextran intestinal permeability test depicted that the intestinal permeability of mice in the ACWP group increased significantly. The gut microbiota analysis in mice depicted that five bacterial strains in the ACWP group, including s_Candidatus_Arthromitus_sp._SFB_mice_Japa, were up-regulated, and nine low-abundant strains, including s_unclassified_g_norank_f_Ruminococcaceae, were down-regulated. The association analysis of gut microbiota and serum factors showed that eight serum biochemical factors were significantly correlated with 11 strains. The results revealed that ACWP could cause disturbance of gut microbiota and its metabolism, change the integrity and permeability of the intestinal tract, destroy the intestinal mucosal immune system, and then cause skin erythema and diarrhea as the primary manifestations.

Walnut green husk ethanol extract improves gut microbiota and their metabolites associated with NLRP3 in non-alcoholic steatohepatitis.

Increasing studies have shown that walnut green husk (WGH) has obvious effects on reducing lipid, resisting oxidation, and protecting the liver. However, the mechanism by which WGH can prevent high-fat diet (HFD)-induced non-alcoholic steatohepatitis (NASH) remains unclear. This study is aimed at investigating the effects of WGH ethanol extract (WGHE) on NLRP3-related biochemical indicators and the diversity and metabolism of gut microbiota in HFD-induced NASH rats. WGHE was administered to HFD-induced NASH rats for 6 weeks. The results showed that WGHE could decrease the levels of blood and liver TC, TG, LDL-C, AST, and ALT and the levels of liver indices, including IL-1ß, IL-6, TNF-α, TGF-ß, FFA, VLDL, caspase-1, ASC, and NLRP3, while it could increase the levels of HDL-C. The pathological damage to liver tissues was significantly reduced. Moreover, WGHE could reduce the Firmicutes/Bacteroidetes (F/B) ratio and the relative abundances of potentially harmful bacteria, such as Lachnospiraceae and Christensenellaceae, and increase that of potentially beneficial bacteria, such as norank_f__Muribaculaceae. These bacteria were associated with NASH and most of them were significantly associated. A total of 23 gut bacteria and 31 metabolites were significantly altered by HFD, which was reversed by WGHE. The common functional pathways, including lipid metabolism and steroid biosynthesis, were identified through the analysis of KEGG metabolic pathways. In addition, the changes in gut microbiota, such as unclassified_f__Lachnospiraceae, unclassified_g__Blautia, and unclassified_g__Desulfovibrio, were associated with the changes in key intestinal metabolites, such as arachidonoyl amine, xanthine, and 25,26-epoxy-1α-hydroxyvitamin D3. In conclusion, WGHE could mitigate HFD-induced NASH in rats by interfering with the NLRP3-related gut microbiota and their metabolites.

Mesenchymal Stem Cell-Derived Extracellular Vesicles in Liver Immunity and Therapy.

Mesenchymal stem cells (MSCs), as the most common cell source for stem cell therapy, play an important role in the modulation of innate and adaptive immune responses and have been widely used in clinical trials to treat autoimmune and inflammatory diseases. Recent experimental and clinical studies have shown that MSC-derived extracellular vesicles (MSC-EVs) can inhibit the activation and proliferation of a variety of proinflammatory cells, such as Th1, Th17 and M1 macrophages, reducing the secretion of proinflammatory cytokines, while promoting the proliferation of anti-inflammatory cells, such as M2 macrophages and Tregs, and increasing the secretion of anti-inflammatory cytokines, thus playing a role in immune regulation and exhibiting immunomodulatory functions. Besides MSC-EVs are more convenient and less immunogenic than MSCs. There is growing interest in the role of MSC-EVs in liver diseases owing to the intrinsic liver tropism of MSC-EVs. In this review, we focus on the immunomodulatory effects of MSC-EVs and summarize the pivotal roles of MSC-EVs as a cell-free therapy in liver diseases, including NAFLD, AIH, acute liver failure, liver fibrosis and hepatic ischemia-reperfusion injury. Moreover, we provide a concise overview of the potential use and limits of MSC-EVs in clinical application.

Diaphragma juglandis extracts modifies the gut microbiota during prevention of type 2 diabetes in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: The diaphragma juglandis (DJ) comes from the wooden septum in the core of Juglans regia L, also known as the walnut septum. In Iranian traditional medicine, walnut distraction wood was widely used in the treatment of diabetes. However, there is a lack of research data on the mechanism of DJ against diabetes. AIM OF THE STUDY: To explore the protective effect of diaphragma juglandis extract (DJE) on type 2 diabetic rats and the hypoglycemic mechanism of DJE. MATERIAL AND METHODS: Supplemented DJE and fed a high-fat diet for five weeks, and then injected low-dose STZ, successfully induced type 2 diabetic rats. Collected rat serum, liver, pancreas and feces to determine the biochemical parameters of serum and liver, analyze the pathological damages of pancreas and liver, and measure the changes of gut microbes in feces. RESULTS: DJE could inhibit the metabolic abnormalities of T2DM by improving insulin resistance, abnormal lipid metabolism, liver damage, oxidative stress, and reducing inflammation. DJE significantly held fasting blood glucose, glycosylated serum protein, serum low density lipoprotein, high density lipoprotein, oral glucose tolerance test, nitric oxide, superoxide dismutase and catalase, serum and liver triglycerides, total cholesterol, aspartate aminotransferase, alanine aminotransferase, malondialdehyde, lipopolysaccharide, fasting insulin and tumor necrosis factor-α and prevented the pathological damage of pancreas and liver. The 16SrRNA gene sequencing results showed that DJE intercepted the disorders of the fecal gut microbes, mainly including Lactobacillaceae, Rikenella, Pygmaiobacter, Oscillospiraceae and Klebsiella. Spearman correlation analysis showed that the changes of gut microbes were closely relative with biochemical parameters. CONCLUSION: DJE might prevent type 2 diabetes and its complications and hold up the disorders of gut microbes.

Long Noncoding RNA and Circular RNA Expression Profiles of Monocyte-Derived Dendritic Cells in Autoimmune Hepatitis.

Autoimmune hepatitis (AIH) is a chronic liver disease caused by disruption of liver immune homeostasis. The effect of dendritic cells (DCs) on the pathogenesis of AIH is not fully understood. Long noncoding RNAs (lncRNAs), circular RNAs (circRNAs), and microRNAs (miRNAs) have been shown to play critical roles in the regulation of cell function. In this study, we analyzed the immunophenotypic characteristics of DCs in the peripheral blood. The percentage of mature DCs was higher in AIH patients than in healthy controls (HCs), and the proportion of mature DCs decreased after treatment. We isolated monocyte-derived DCs (moDCs) from the peripheral blood, obtained whole RNA-sequencing (RNA-seq) data for the moDCs from the two groups, and identified differentially expressed (DE) lncRNAs, circRNAs, miRNAs and mRNAs. In addition, we performed Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses for the DE mRNAs and constructed competing endogenous RNA (ceRNA) networks. ENST00000543334, hsa_circ_0000279, and hsa_circ_0005076 were selected and validated by RT-qPCR. These results provide a possible molecular mechanism of DCs in the pathogenesis of AIH and identify some potential therapeutic targets.

Glycolipid metabolism and liver transcriptomic analysis of the therapeutic effects of pressed degreased walnut meal extracts on type 2 diabetes mellitus rats.

Walnut meal (WM) is rich in polyphenols which exhibit multiple therapeutic effects. The purpose of this study was to investigate the therapeutic effects of walnut meal extracts (WMP) on glycolipid metabolism and liver transcriptomics in T2DM rats. A T2DM rat model was established by using a high-fat diet combined with streptozotocin. A 5-week WMP therapy showed the effects of decreasing water intake, excretion, fasting blood glucose, fasting insulin, and insulin resistance, increasing ß-cell function and insulin sensitivity index; meanwhile regulating dysfunctional lipid metabolism and reducing inflammation; improving body weight, oral glucose tolerance test and insulin sensitivity; and increasing the activities of SOD and CAT while decreasing the MDA levels in the liver and serum of T2DM rats. Moreover, 10 key differentially expressed genes were identified by RNA-seq, including Gck, RT1-Ba, Fasn, Slc13a3, Cd74, Jun, Cyp4a1, Myh7b, Plin3, and Got1, and they were highly potentially related to glycolipid metabolism. Our results suggested that WMP exhibited the anti-diabetic effect and could regulate glycolipid metabolism in T2DM rats. This finding might assist in identifying potential therapeutic targets for T2DM prevention and intervention.

Protective effect of walnut on d-galactose-induced aging mouse model.

OBJECTIVE S: Accumulating evidence has suggested that oxidative stress and apoptosis are involved in the aging process. d-galactose (gal) has been reported to cause symptoms of aging in mice, accompanied by liver and brain injuries. Our present work was to study the potential antioxidative and anti-apoptotic effects of walnut and to explore how these effects act on mice in a d-gal-induced aging model. MATERIALS AND METHODS: Aging mice were induced by subcutaneous injection of d-gal (200 mg kg-1 d-1 for 8 weeks). Walnut samples were simultaneously administered to the d-gal-induced aging mice once daily by intragastric gavage. Finally, body weight, organ index, cognitive function, levels of antioxidative enzymes, and liver function were monitored. RESULTS: The kernel pellicles of walnut could not only improve the learning and memory ability, and the organ index, but also significantly decrease body weight and normalize the levels of activity of antioxidative enzymes in aging mice. Further, the walnut seed coat would protect damages of hippocampus and liver in aging mice. HIGHLIGHTS: In the current study, we investigated the effects of walnut kernels and walnut seed coats (WSCs) on d-galactose-induced aging mice. WSC was firstly found to have beneficial effects on d-gal-treated mouse's brain with learning and memory impairment, which probably through the underlying mechanism reduces oxidative damage and limits neuroinflammation. In addition, WSC had a protective effect on liver damage in d-galactose sensing mice.

Optimization of simultaneous microwave/ultrasonic-assisted extraction of phenolic compounds from walnut flour using response surface methodology.

CONTEXT: Walnut is a traditional food as well as a traditional medicine recorded in the Chinese Pharmacopoeia; however, the large amounts of walnut flour (WF) generated in walnut oil production have not been well utilized. OBJECTIVE: This study maximized the total polyphenolic yield (TPY) from the walnut flour (WF) by optimizing simultaneous ultrasound/microwave-assisted hydroalcoholic extraction (SUMAE). MATERIALS AND METHODS: Response surface methodology was used to optimize the processing parameters for the TPY, including microwave power (20-140 W), ultrasonic power (75-525 W), extraction temperature (25-55 °C), and time (0.5-9.5 min). The polyphenol components were analysed by LC-MS. RESULTS: A second-order polynomial model satisfactorily fit the experimental TPY data (R2 = 0.9932, P < 0.0001 and Radj2 = 0.9868). The optimized quick extraction conditions were microwave power 294.38 W, ultrasonic power 93.5 W, temperature 43.38 °C and time 4.33 min, with a maximum TPY of 34.91 mg GAE/g, which was a rapid extraction. The major phenolic components in the WF extracts were glansreginin A, ellagic acid, and gallic acid with peak areas of 22.15%, 14.99% and 10.96%, respectively, which might be used as functional components for health food, cosmetics and medicines. DISCUSSION AND CONCLUSION: The results indicated that walnut flour, a waste product from the oil industry, was a rich source of polyphenolic compounds and thus could be used as a high-value functional food ingredient.

Crystal structure of (2E,3E)-N (2),N (3)-bis-(3-ethyl-[1,1'-biphen-yl]-4-yl)butane-2,3-di-imine.

In the title compound, C32H32N2, synthesized by the con-densation reaction of 2-ethyl-4-phenyl-aniline and 2,3-butane-dione, the conformation about the C=N bonds is E and the substituted biphenyl units are trans to one another. In the two biphenyl ring systems, the planes of the two rings are inclined to one another by 25.25 (19) and 28.01 (19)°. The planes of the ethyl-substituted benzene rings are inclined to one another by 20.23 (19)° and to the mean plane of the butane-2,3-di-imine unit [maximum deviation = 0.014 (4) Å] by 83.19 (19) and 63.38 (19)°. In the crystal, mol-ecules are linked by C-H⋯π inter-actions, forming sheets lying parallel to (101).

Crystal structure of 2,3-bis-[(4-tert-butyl-2,6-di-methyl-phen-yl)imino]-butane.

The title compound, C28H40N2, was obtained from the condensation reaction of 4-tert-butyl-2,6-di-methyl-aniline and butane-2,3-dione. The mol-ecule lies on an inversion centre. The C=N bond has an E conformation. The plane of the benzene ring is almost perpendicular to the 1,4-di-aza-butadiene mean plane [dihedral angle = 89.8 (9)°].

Interaction of gallic acid with trypsin analyzed by spectroscopy.

The interactions between trypsin and gallic acid (GA) were investigated by means of fluorescence spectroscopy, UV-vis absorption spectroscopy, resonance light scattering (RLS) spectroscopy, synchronous fluorescence spectroscopy, and enzymatic inhibition assay. It was found that GA can cause the fluorescence quenching of trypsin during the process of formation of GA-trypsin complex, resulting in inhibition of trypsin activity (IC50 = 3.9 × 10-6 mol/L). The fluorescence spectroscopic data showed that the quenching efficiency can reach about 80%. The binding constants were 1.9371 × 104 L/mol, 1.8192 × 104 L/mol, and 1.7465 × 104 L/mol at three temperatures, respectively. The thermodynamic parameters revealed that hydrogen bonds, van der Waals, hydrophobic, and electrostatic interactions were involved in the binding process of GA to trypsin. Molecular modeling studies illustrated a specific display of binding information and explained most of the experiment phenomena. The microenvironments of tryptophan and tyrosine residue in trypsin were changed by the GA. Results indicated that GA was a strong quencher and inhibitor of trypsin.

Discrimination of Rhizoma Gastrodiae (Tianma) using 3D synchronous fluorescence spectroscopy coupled with principal component analysis.

Rhizoma Gastrodiae (Tianma) of different variants and different geographical origins has vital difference in quality and physiological efficacy. This paper focused on the classification and identification of Tianma of six types (two variants from three different geographical origins) using three dimensional synchronous fluorescence spectroscopy (3D-SFS) coupled with principal component analysis (PCA). 3D-SF spectra of aqueous extracts, which were obtained from Tianma of the six types, were measured by a LS-50B luminescence spectrofluorometer. The experimental results showed that the characteristic fluorescent spectral regions of the 3D-SF spectra were similar, while the intensities of characteristic regions are different significantly. Coupled these differences in peak intensities with PCA, Tianma of six types could be discriminated successfully. In conclusion, 3D-SFS coupled with PCA, which has such advantages as effective, specific, rapid, non-polluting, has an edge for discrimination of the similar Chinese herbal medicine. And the proposed methodology is a useful tool to classify and identify Tianma of different variants and different geographical origins.

A preliminary metagenomic study of puer tea during pile fermentation.

BACKGROUND: Up to now, there has been no report on the taxonomic and functional analysis of the microbial community in fermenting puer tea by pyrosequencing. In this study, metagenomic pyrosequencing was first used in fermenting puer tea to delineate a relatively comprehensive overview of the microbial taxonomy while also preliminarily characterising the functional ontologies of microbial genes present in puer tea pile fermentation. RESULTS: A total of 251 738 pyrosequencing reads (9197 contigs and 145 402 singletons) were generated by pyrosequencing. Taxonomic analysis revealed three dominant bacterial phyla, Actinobacteria (30.08%), Proteobacteria (24.47%) and Firmicutes (20.23%), and one dominant eukaryotic phylum, Ascomycota (15.21%) [corrected]. A total of 58 664 hits were categorised into 28 functional subsystems based on the SEED database. Moreover, two categories, 'metabolism of terpenoids and polyketides' and 'biosynthesis of other secondary metabolites', were selectively analysed and 69 enzyme genes were presented in 16 pathways. CONCLUSION: The dominant microbes of puer tea fermentation were bacteria in the present study, and yeasts rather than moulds accounted for the overwhelming majority of Eukaryota. The analysis of functional genes and metabolic pathways will be helpful for further study of the mechanism of puer tea fermentation at molecular level.

Comparison of laxative and antioxidant activities of raw, processed and fermented Polygoni Multiflori radix.

AIM: To observe the anti-oxidative activity and adverse laxative effect of raw, traditional processed and fermented products of Polygoni Multiflori Radix (PMR), and furthermore, to evaluate the fermentation method used in the processing procedure of PMR. METHODS: In vitro ferric reducing antioxidant power (FRAP) assay was carried out to evaluate the anti-oxidative activity. Modulation of normal defecation and effect on gastrointestinal motility in mice were carried out to investigate their adverse laxative effect. RESULTS: Fermented PMR induced less severe laxative adverse effect than Polygoni Multiflori Radix Praeparata (PMRP). PMR fermented with Rhizopus sp. (FB) could modulate the defecation significantly. The gastrointestinal motility was inhibited by PMRP and PMR fermented with Rhizopus oryzae (FA). FA and FB showed better antioxidant activity than PMRP in 50% and 95% ethanol group. Contents of 2, 3, 5, 4'-tetrahydroxy-stilbene-2-O-ß-D-glucoside (TSG) were reduced significantly after traditional processing but maintained after fermentation. Emodin and physcion were increased after traditional processing and fermented with Rhizopus oryzae. CONCLUSION: All processing procedure, including fermentation, might reduce its anti-oxidative activity. However, most of the processed products could lessen the adverse effect on gastrointestinal tract compared to PMR. Fermentation with Rhizopus oryzae was considered as a promising processing method of PMR.

Evaluation of Cynanchum otophyllum glucan sulfate against human immunodeficiency virus and herpes simplex virus as a microbicide agent.

OBJECTIVE: The root of Cynanchum otophyllum-also known as Qing Yang Sheng-is a traditional ethnical Chinese medicine. The objective of this study was to evaluate in vitro activities and safety of C. otophyllum glucan sulfate (PS20) against Human Immunodeficiency Virus (HIV) and Herpes Simplex Virus (HSV). MATERIALS AND METHODS: Anti-HIV activity was detected with syncytial formation assay and quantitative P24 Enzyme-Linked Immunosorbent Assay (ELISA). Anti-HSV activity was detected with plaque reduction assay; cytotoxicity was tested with MTT colorimetric assay; and anti-bacterial activity was tested with microdilution method. Anti-HIV mechanism was investigated with fusion inhibition, time of addition, and pretreatment. RESULTS: The 50% Inhibition Concentration (IC(50)) of PS20 for HIV-1(IIIB), HIV-(Ada-M), HIV-1(Bal), HSV-I, and -II were 0.26 ± 0.02 mM, 0.46 ± 0.02 mM, 0.90 ± 0.04 mM, 3.45 ± 0.85 µM, and 0.70 ± 0.22 mM, respectively. Selectivity Indices (SI) were 653, 50, 39, 85, and 362, respectively. Studies on anti-HIV mechanism of PS20 showed that the target molecule should be the envelope protein. The 50% Cytotoxicity Concentrations (CC(50)) of PS20 for HeLa and ME-180 cell lines and human foreskin fibroblast cells was more than 70 µM. The Minimum Inhibitory Concentration (MIC) for vaginal lactobacilli was more than 1000 µM. CONCLUSION: PS20 possesses anti-HIV and HSV effect and low cytotoxicity to epithelium cells and vaginal lactobacilli. It may be considered as a potential microbicide agent for further investigation.