Ameliorative Effect of Areca Nut Polyphenols on Adverse Effects Induced by Lipopolysaccharides in RAW264.7 Cells.

In Asian regions, areca nuts are tropical fruits that are extensively consumed. The areca nut contains a lot of polyphenols and its safety is unknown. In this research, we investigated the effects of lipopolysaccharides (LPS) and areca nut polyphenols (ANP) on normal RAW264.7 cells. The results showed that LPS stimulated adverse effects in normal cells by affecting cytokine production. The GO analysis results mainly affected DNA repair, cell division, and enzyme activities. In the KEGG analysis results, the NOD-like receptor signaling pathway, which is related to NF-κB, MAPK, and the pro-inflammatory cytokines, is the most significant. In the protein-protein interaction network (PPI) results, significant sub-networks in all three groups were shown to be related to cytokine-cytokine receptor interaction. Collectively, our findings showed a comprehensive understanding of LPS-induced toxicity and the protective effects of ANP by RNA sequencing.

Assessment of Areca Nut Bioactivities in Western Diet-Induced Mice NAFLD Model.

The areca nut is often consumed as a chewing food in the Asian region. Our previous study revealed that the areca nut is rich in polyphenols with high antioxidant activity. In this study, we further assessed the effects and molecular mechanisms of the areca nut and its major ingredients on a Western diet-induced mice dyslipidemia model. Male C57BL/6N mice were divided into five groups and fed with a normal diet (ND), Western diet (WD), WD with areca nut extracts (ANE), areca nut polyphenols (ANP), and arecoline (ARE) for 12 weeks. The results revealed that ANP significantly reduced WD-induced body weight, liver weight, epididymal fat, and liver total lipid. Serum biomarkers showed that ANP ameliorated WD-enhanced total cholesterol and non-high-density lipoprotein (non-HDL). Moreover, analysis of cellular signaling pathways revealed that sterol regulatory element-binding protein 2 (SREBP2) and enzyme 3-hydroxy-3-methylglutaryld coenzyme A reductase (HMGCR) were significantly downregulated by ANP. The results of gut microbiota analysis revealed that ANP increased the abundance of beneficial bacterium Akkermansias and decreased the abundance of the pathogenic bacterium Ruminococcus while ARE shown the opposite result to ANP. In summary, our data indicated that areca nut polyphenol ameliorated WD-induced dyslipidemia by increasing the abundance of beneficial bacteria in the gut microbiota and reducing the expressions of SREBP2 and HMGCR while areca nut ARE inhibited this improvement potential.

UPLC-QQQ-MS/MS-based widely targeted metabolomic analysis reveals the effect of solid-state fermentation with Eurotium cristatum on the dynamic changes in the metabolite profile of dark tea.

Eurotium cristatum is the predominant fungus and key contributor to the characteristics of post-fermented Fu brick tea (FBT) during manufacturing. In this study, the influence of solid-state fermentation (SSF) with E. cristatum on the chemical profile dynamic changes of dark tea was investigated. Results indicated that total phenolics, flavonoids, theaflavins, thearubigins, and galloyl catechins consistently decreased, degalloyl catechins and gallic acid increased in the initial stage of fermentation and decreased after long-term fermentation, and theabrownins continually increased. UPLC-QQQ-MS/MS-based widely targeted metabolomic analysis revealed that the metabolites of dark tea processed by SSF with E. cristatum were drastically different from the raw material. A total of 574 differential metabolites covering 11 subclasses were detected in the whole SSF of dark tea, and the most drastic changes occurred in the middle stage. Phenolic acids and flavonoids were the two major classes of differential metabolites. A series of reactions such as degradation, glycosylation, deglycosylation, methylation, and oxidative polymerization occurred during SSF. Overall, SSF with E. cristatum greatly influenced the metabolites of dark tea, which provided valuable insights that E. cristatum is critical in forming the chemical constituents of FBT.

Effects of post-treatment electroacupuncture on ventricular monophasic action potential and cardiac function in a rat model of ischemia/reperfusion injury.

BACKGROUND: To determine the effects of post-treatment electroacupuncture (EA) on the electrophysiological properties of ventricular muscle in rats with ischemia/reperfusion (IR) injury. METHODS: Male Sprague-Dawley (SD) rats were randomly assigned into sham-operated (SH), IR and IR + EA groups (n = 8 each). The IR model was generated by ligation of the left anterior descending (LAD) coronary artery for 30 min. After establishing the IR model, EA was administered at PC6 for 30 min while opening the coronary artery and allowing reperfusion for 30 min. Heart rate (HR), mean arterial pressure and monophasic action potential (MAP) of cardiac muscle in the outer membrane of the antetheca of the left ventricle before coronary artery ligation (T0), after coronary artery ligation for 30 min (T1) and after reperfusion for 30 min (T2) were recorded. At the same time, ventricular electrophysiological parameters including ventricular effective refractory period (ERP), conduction velocity (CV) and ventricular fibrillation threshold (VFT) were measured. Then, the cardiac function and the levels of creatine kinase-muscle/brain (CK-MB) and cardiac troponin I (cTnI) were monitored. Based on these data, monophasic action potential amplitude (MAPA), the maximum depolarization velocity (Vmax) and the MAP durations at 50% and 90% repolarization (MAPD50 and MAPD90) were calculated to determine the incidence of arrhythmia during reperfusion. RESULTS: Compared with the SH group, the IR group showed an obviously decreased HR as well as reduced mean arterial pressure, Vmax, CV, ERP and MAPA. All indices of cardiac function except left ventricular end-diastolic pressure (LVEDP) decreased (i.e. ventricular systolic pressure (LVSP), left ventricular ejection fraction (LVEF), fractional shortening (FS) and rate of the ventricular pressure rise/drop (±dp/dt)). Furthermore, the MAPD50 and MAPD90 were prolonged, and the levels of CK-MB and cTnI increased (p < 0.05). In comparison to the IR group, HR and the mean arterial pressure were increased. All indices of cardiac function except LVEDP increased (LVSP, LVEF, FS and ±dp/dt). Vmax, CV, ERP and MAPA were also increased in the IR + EA group. However, MAPD50 and MAPD90 were distinctly shortened, and the levels of CK-MB and cTnI decreased (p < 0.05). There were no statistically significant differences in VFT between the three groups (p > 0.05). CONCLUSION: EA post-treatment can relieve prolongation of repolarization and slowed depolarization of ventricular muscle during IR, thus decreasing the rate of incidence of reperfusion arrhythmia.

Metabolite profiling, antioxidant and α-glucosidase inhibitory activities of buckwheat processed by solid-state fermentation with Eurotium cristatum YL-1.

Buckwheat was processed by solid-state fermentation (SSF) with the probiotic fungal strain Eurotium cristatum YL-1. The effects of SSF on the phytochemical content, as well as the antioxidant and α-glucosidase inhibitory activities, on buckwheat were revealed. Metabolite differences between non-fermented buckwheat (BW) and E. cristatum fermented buckwheat (FBW) were investigated by LC-MS/MS-based untargeted metabolomics. Results showed that 103 and 68 metabolites remarkably differed between BW and FBW in positive and negative ionization modes, respectively. Most phenolic compounds and alkaloids were significantly up-regulated during SSF. Hydrolytic enzymes (i.e., ß-glucosidase, α-amylase, protease, and cellulase) were produced by the filamentous fungus E. cristatum during SSF. In vitro spectrophotometric assays demonstrated that the total phenolics content, ferric reducing antioxidant power, reducing power, scavenging activities of DPPH radical and ABTS+, and α-glucosidase inhibitory activity of buckwheat were considerably enhanced after processing by SSF with E. cristatum. Additionally, solvents with different polarities significantly influenced the antioxidant and α-glucosidase inhibitory activities of buckwheat extracts. Our study indicated that processing by SSF with E. cristatum can greatly improve the phytochemical components of buckwheat and consequently contribute to its antioxidant and α-glucosidase inhibitory activities. SSF with E. cristatum is an innovative method for enhancing the health-promoting components and bioactivities of buckwheat.

Effect of Hyperbaric Oxygen on Tissue Damage and Expression of Adhesion Molecules and C3 in a Rat Model of Renal Ischemia-Reperfusion Injury After Kidney Transplantation.

BACKGROUND The aim of this study was to investigate the protective effect and mechanism of hyperbaric oxygen (HBO) in a rat model of renal ischemia-reperfusion injury following kidney transplantation. MATERIAL AND METHODS Sprague Dawley rats were randomly divided into 3 groups (n=18): sham group, kidney transplantation group, and HBO treatment group. Six rats in each group were sacrificed at 1, 3, and 5 hours after reperfusion, and serum and renal tissue were then collected. The serum creatinine levels and histopathological changes of the renal tissue were detected. ICAM-1, VCAM-1, and C3 expression levels were also detected by immunohistochemical staining or real-time polymerase chain reaction. RESULTS Renal function was damaged in the kidney transplantation group and the HBO treatment group compared with sham group (P<0.05). Renal histopathological changes, including tubular cell swelling, tubular dilatation, and hyaline casts, were remarkably reduced in the HBO treatment group compared to the kidney transplantation group. In the immunohistochemical examination, the expression levels of ICAM-1, VCAM-1, and C3 were obviously increased in the kidney transplantation group and the HBO treatment group; moreover, the levels in the HBO treatment group were significantly lower than in the kidney transplantation group (P<0.05). In addition, the ICAM-1 and C3 mRNA levels were increased in the kidney transplantation group and HBO treatment group, but the levels of in the HBO treatment group them were significantly decreased compared to the kidney transplantation group that at 3 and 5 hours after reperfusion (P<0.05). CONCLUSIONS HBO treatment exerted a protective effect on renal function through inhibition of adhesion molecule overexpression and complement system activation in a rat model of renal ischemia-reperfusion injury after kidney transplantation.

A comparison for the effects of raw, smoked, and smoked and brined areca nut extracts on the immune and inflammatory responses in the Kunming mice.

Chewing of areca nuts is quite popular in various regions worldwide. Previous studies have demonstrated the pharmacological and toxicological effects of fresh areca nuts. However, processed areca nuts, which are popular in the Hunan province of China, have not been extensively studied for its biological effect. This study aimed at investigating the impact of the acrea nut extracts (ANE) prepared from the raw material, the semi-product, and the final product on the immune system and inflammation-related markers in the Kunming mice. The mice were assigned to seven different groups and administered different ANE at two concentrations (1X and 5X) for four weeks. Total body weight gain and organ coefficient of the liver, spleen, and kidney, as well as the immune system and inflammation-related markers were evaluated. The results revealed that processed areca nuts have a much milder effect on the mice immune system and some inflammatory markers than fresh areca nut in the Kunming mice. PRACTICAL APPLICATIONS: Chewing various forms of areca nuts is popular in China, Southeast Asia, and other regions. People from Hunan, China prefer to chew a processed areca nut, which has rarely been studied. This manuscript explores the effects of three kinds of areca nut extracts on the immune system- and inflammation-related indicators in Kunming mice. The obtained results revealed that processed areca nuts had significantly milder effects than the raw nut/nut extract, particularly on the body weight, immune responses, and inflammatory markers. The results of the present study provide some new directions for the areca nut industry and raise public awareness for the undesirable effects of areca nuts.

Semi-continuous production of high-activity pectinases by immobilized Rhizopus oryzae using tobacco wastewater as substrate and their utilization in the hydrolysis of pectin-containing lignocellulosic biomass at high solid content.

In this study, highly reactive endo- and exo-polygalacturonases (PGs) were produced from the tobacco industry wastewater using immobilized Rhizopus oryzae. Compared with free cells, immobilized cells increased enzyme activity 2.8-fold and reduced production time to 24h by shake-flask production. Moreover, the immobilized cells enabled the semi-continuous production of enzymes through repeated-batch mode for seven consecutive cycles in a scale-up bioreactor. During the first five cycles, the average endo-PG and exo-PG activities reached 307.5 and 242.6U/ml, respectively. The addition of crude enzyme for the hydrolysis of pectin-containing lignocellulosic biomass under high-gravity conditions increased glucose release 4.2-fold (115.4 vs. 29.0g/L), compared with hydrolysis using cellulase alone. This process achieves the efficient production of pectin-degrading enzymes, provides a cost-effective method for tobacco wastewater treatment, and offers the possibility to obtain fermentable sugars with high-titer from pectin-containing lignocellulosic biomass, which has important potential for the commercial production of bio-fuels.

Effects of Fuzhuan Brick-Tea Water Extract on Mice Infected with E. coli O157:H7.

Fuzhuan brick-tea extract (FBTE) affects the physiology of mice infected with Escherichia coli O157:H7. For 10 consecutive days, 0.05, 0.5, and 1.0 g/mL FBTE was administered intragastrically to three groups of infected Kunming mice, and changes in immunological function, hematology, and histopathology were examined. The results revealed upregulation of platelets, total protein, and albumin along with downregulation of serum triglycerides, aspartate aminotransferase, creatinine, and urea nitrogen in FBTE-treated mice. Histological sections of stomach, kidney, duodenum, ileum, and colon suggested that infected mucous membranes could be rehabilitated by low- and high-dose FBTE and that inflammation was alleviated. Similarly, increased thymic function in mice treated with middle- and high-dose FBTE led to elevated serum hemolysin antibody titer and increased CD4+ and CD8+ T cells, as indicated by CD4+ and CD8+ expression on intestinal mucosa. Monocyte and macrophage function was improved by three FBTE dosages tested. Colonic microbiota analysis by denaturing gradient gel electrophoresis (DGGE) revealed characteristic bands in infected mice treated with middle- and high-dose FBTE and increased species diversity in Lactobacillus, Bacteroides, and Clostridium cluster IV. These results suggest that FBTE may protect kidney and liver of mice infected with E. coli O157:H7, improve immune function, and regulate the colonic microbiota.

Fungal community associated with fermentation and storage of Fuzhuan brick-tea.

Chinese Fuzhuan brick-tea is a unique microbial fermented tea characterized by a period of fungal growth during its manufacturing process. The aim of the present study was to characterize, both physicochemically and microbiologically, traditional industrial production processes of Fuzhuan brick-tea. Fermenting tea samples were collected from the largest manufacturer. Physicochemical analyses showed that the low water content in the tea substrates provided optimal growth conditions for xerophilic fungi. The fungal communities existing in tea materials, fermenting tea, and stored teas were monitored by polymerase chain reaction (PCR)-denaturing gradient gel electrophoresis (DGGE) targeting the D1 region of the 26S rRNA genes, followed by sequencing of the amplicons. Results revealed that the microorganisms were from, or closely related to, the genera Eurotium, Debaryomyces, Aspergillus, Verticillium, Pichia, Pestalotiopsis, Rhizomucor and Beauveria. This is the first report of Debaryomyces participating in the processing of Fuzhuan brick-tea. We concluded that the dominant genera Eurotium, Debaryomyces and Aspergillus are beneficial fungi associated with the fermentation of Fuzhuan brick-tea. The genus Beauveria was present in the stored Fuzhuan brick-tea, which may help protect tea products from insect spoilage. The remaining four genera were of minor importance in the manufacturing of Fuzhuan brick-tea. The predominant Eurotium species, a strain named Eurotium sp. FZ, was phenotypically and genotypically identified as Eurotium cristatum. High performance thin layer chromatography analysis of anthraquinones showed that emodin existed in all the dark tea samples, but physcion was only detectable in the tea fermented by E. cristatum. The PCR-DGGE approach was an effective and convenient means for profiling the fungal communities in Fuzhuan brick-tea. These results may help promote the use of microbial consortia as starter cultures to stabilize and improve the quality of Fuzhuan brick-tea products.

Study on degradation performance of star-shaped polymer of ESO and lactide / 生物医学工程学杂志

Star-shaped polymer of epoxidized soybean oil (ESO) and lactide is a new material. Its degradation function has an importance to its applications. There are many factors affecting its degradation. This paper is mainly aimed at investigation in which the surface morphology, pH, molecular weight and weight loss rate, branching ESO-lactide star polymers the degree of change were studied. It was found through the experiments that, the changing rates of the branching ESO-lactide star polymers were slower than the degree of PDLLA change compared to the surface morphology, pH, molecular weight and weight loss rate. This has a guiding significance to the further application of ESO-lactide.