Metagenomic analysis revealed the evolution of microbial communities, metabolic pathways, and functional genes in the heterotrophic nitrification-aerobic denitrification process under La3+ stress.

Trivalent lanthanum (La3+) exists widely in ammonia nitrogen (NH4+-N) tailing water from ionic rare earth mines; however, its effect on heterotrophic nitrification-aerobic denitrification (HN-AD) is unknown, thereby limiting the application of the HN-AD process in this field. In this study, we conducted an HN-AD process using a sequencing batch reactor (5 L) that was continuously operated to directly treat acidic (NH4)2SO4 wastewater (influent NH4+-N concentration of approximately 110 mg/L and influent pH of 5) containing different La3+ concentrations (0-100 mg/L). The NH4+-N removal efficiency of the reactor reached 98.25 % at a La3+ concentration of 100 mg/L. The reactor was in a neutral-to-alkaline environment, which favored La3+ precipitation and complexation. Metagenomic analysis revealed that the relative abundance of Thauera in the reactor remained high (88.62-92.27 %) under La3+ stress. The relative abundances of Pannonobacter and Hyphomonas significantly increased, whereas that of Azoarcus significantly decreased. Metabolic functions in the reactor were mainly contributed by Thauera, and the abundance of metabolic functions under low La3+ stress (≤5 mg/L) significantly differed from that under high La3+ stress (≥10 mg/L). The relative abundance of ammonia assimilation-related genes in the reactor was high and significantly correlated with ammonia removal. However, traditional ammonia oxidation genes were not annotated, and unknown ammonia oxidation pathways may have been present in the reactor. Moreover, La3+ stimulated amino acid biosynthesis and translocation, the citrate cycle, sulfur metabolism, and oxidative phosphorylation and promoted the overproduction of extracellular polymeric substances, which underwent complexation and adsorbed La3+ to reduce its toxicity. Our results showed that the HN-AD process had a strong tolerance to La3+, stable NH4+-N removal efficiency, the potential to recover La3+, and considerable application prospects in treating NH4+-N tailing water from ionic rare earth mines.

Performance changes in the anammox process under the stress of rare-earth element Ce(III) and the evolution of microbial community and functional genes.

The high Ce(III) content in ionic rare-earth tailings wastewater has hindered the application of anammox process in this field. Here, the effect of Ce(III) on the performance of anammox processes was investigated, and the evolution of microbial communities and functional genes was explored using metagenomic sequencing. The results showed that the reactor nitrogen removal rate decreased when the Ce(III) concentration reached 25 mg/L, although ammonia nitrogen removal (92.31%) and nitrogen removal efficiency (81.33%) remained at a high level; however, both showed a significant decreasing trend. The relative abundance of anammox bacteria increased continuously from P1-P5, reaching 48.81%, whereas the relative abundance of Candidatus jettenia reached 33.71% at P5, which surpassed that of Candidatus brocadia as the most abundant anammox bacteria, and further analysis of functional genes and metabolic pathways revealed that Candidatus brocadia was richer in biochemical metabolic genes, whereas Candidatus jettenia had richer efflux genes.

MOFs-derived CuO-Fe3O4@C with abundant oxygen vacancies and strong Cu-Fe interaction for deep mineralization of bisphenol A.

A novel CuO-Fe3O4 encapsulated in the carbon framework with abundant oxygen vacancies (CuO-Fe3O4@C) was successfully prepared by thermal conversion of Cu(OAc)2/Fe-metal organic framework. The as-prepared catalyst exhibited excellent peroxymonosulfate (PMS) activation performance, good recyclability and fast magnetic separation. Under optimal conditions, the added BPA (60 mg/L) could be completely removed by CuO-Fe3O4@C/PMS system within 15 min with the degradation rate constant (k) of 0.32 min-1, being 10.3 and 246.2 times that in CuO/PMS (0.031min-1) and Fe3O4/PMS (0.0013 min-1) system. A deep mineralization rate of BPA (＞80%) was achieved within 60 min. The results demonstrated the synergistic effect of bimetallic clusters, oxygen vacancies and carbon framework was a key benefit for the exposure of more active sites, the electron donor capacity and the mass transfer of substrates, thereby promoting the decomposition of BPA. Capture experiments and EPR indicated that 1O2 was the predominant reactive oxygen species (ROSs). The degradation routes of BPA and the activation mechanism of PMS were proposed. This study offers an opportunity to develop promising MOFs-derived hybrid catalysts with tailored structures and properties for the practical application of SR-AOPs.

The opening of mitochondrial permeability transition pore (mPTP) and the inhibition of electron transfer chain (ETC) induce mitophagy in wheat roots under waterlogging stress.

Mitochondria are crucial for the regulation of intracellular energy metabolism, biosynthesis, and cell survival. And studies have demonstrated the role of mitochondria in oxidative stress-induced autophagy in plants. Previous studies found that waterlogging stress can induce the opening of mitochondrial permeability transition pore (mPTP) and the release of cytochrome c in endosperm cells, which proved that mPTP plays an important role in the programmed cell death of endosperm cells under waterlogging stress. This study investigated the effects of the opening of mPTP and the inhibition of ETC on mitophagy in wheat roots under waterlogging stress. The results showed that autophagy related genes in the mitochondria of wheat root cells could respond to waterlogging stress; waterlogging stress led to the degradation of the characteristic proteins cytochrome c and COXII in the mitochondria of root cells. With the prolongation of waterlogging time, the protein degradation degree and the occurrence of mitophagy gradually increased. Under waterlogging stress, exogenous mPTP opening inhibitor CsA inhibited mitophagy in root cells and alleviated mitophagy induced by flooding stress, while exogenous mPTP opening inducer CCCP induced mitophagy in root cells; exogenous mPTP opening inducer CCCP induced mitophagy in root cells. The electron transfer chain inhibitor antimycin A induces mitophagy in wheat root cells and exacerbates mitochondrial degradation. In conclusion, waterlogging stress led to the degradation of mitochondrial characteristic proteins and the occurrence of mitophagy in wheat root cells, and the opening of mPTP and the inhibition of ETC induced the occurrence of mitophagy.

Removal of ammonia nitrogen from residual ammonium leaching solution by heterotrophic nitrification-aerobic denitrification process.

The aftermath of mining weathered crust elution-deposited rare earth ore produces a large amount of residual ammonium leaching solution, which causes ammonia and nitrogen pollution to the mine site. Recently, denitrification by heterotrophic nitrification-aerobic denitrification (HN-AD) bacteria has attracted much attention. However, limited studies exist regarding the denitrification process of HN-AD bacteria. In this study, we combined four strains of HN-AD bacteria, Pseudomonas fulva K3, Pseudomonas mosselii K17, Klebsiella oxytoca A12, and Enterobacter hormaechei A16, obtained from rare earth element leaching sites, to select the best microbial consortium for ammonia nitrogen removal. We designed an ammonia removal process applicable to HN-AD bacteria to directly remove ammonia nitrogen from acidic leaching solutions. The experimental results demonstrated that the most efficient microbial consortium for ammonia nitrogen removal to be K3 + K17 + A16, with a removal efficiency of 89.68% for 8 h. In this process, considering the influencing factors of the ammonia removal process, the larger the influent flow rate and influent ammonia nitrogen concentration, the greater the ammonia nitrogen accumulation and pH decrease in the reactor. In consecutive multi-batch experiments, the ammonia removal process was used to remove ammonia nitrogen, at concentrations of 100-600 mg/L, from the simulated leaching solution at pH 4-7, whereby the effluent ammonia nitrogen concentration was lower than 15 mg/L. The results demonstrate that the ammonia removal process is highly feasible and stable. These findings will provide new ideas for the application of HN-AD bacteria and new methods for the removal of ammonia nitrogen from acidic leaching solutions.

Dietary yeast culture alleviates intestinal-hepatic damage related to TLR2-MyD88-NF-κB signaling pathway and antioxidant capability in Pseudobagrus ussuriensis.

Yeast culture (YC), as a member of probiotics family is a natural product produced from yeast fermentation, affects of improving immunity. However, the intestine and liver injury and immunosuppression mechanism caused by SBA in fish are unclear and more functions of YC supplement in the diet need to be developed. Soybean agglutinin (SBA) is an anti-nutritional factor in soybean and leads to growth-inhibitory effect in feeding of the high proportion of soybean meal replacing fish meal (FM). Therefore, one hundred and thirty-five Pseudobagrus ussuriensis (6.5 ± 0.27 g) were randomly selected and divided into three groups (Control, SBA and YC+SBA groups). For the model, fish were fed with 2% YC for 8 weeks and then given intragastric administration of 0.2-mL SBA solution for 20 days. The results showed that SBA damaged the immune and antioxidant capacity, causing an inflammatory reaction, leading to abnormal expression of cytokines in the intestine and liver of Pseudobagrus ussuriensis. YC could effectively attenuate intestinal and liver damage and downregulate the TLR2/MyD88/NF-κB signaling pathway and suppress oxidative stress in Pseudobagrus ussuriensis. Besides, YC had obvious immune advantage, which could improve the immune ability. In summary, these results showed that YC could reduce immunosuppression and intestinal-liver injury by inhibiting the TLR2/MyD88/NF-κB signal pathway and oxidative stress induced by SBA. This study provided some explanations for the problems of fish diet caused by anti-nutritional factors from soybean meal and provided a theoretical basis for the function development of YC in aquaculture.

Isolation and Nitrogen Removal Efficiency of the Heterotrophic Nitrifying-Aerobic Denitrifying Strain K17 From a Rare Earth Element Leaching Site.

K17, an indigenous and heterotrophic nitrifying-aerobic denitrifying bacterium, was isolated from the soil of a weathered crust elution-deposited rare earth ore leaching site in Longnan County, China. Strain K17 was identified as Pseudomonas mosselii. In this study, the morphological characteristics of strain K17 were observed and the optimal ammonia nitrogen removal conditions for the strain were studied using a single-factor experiment. Key enzyme activities were determined, and we also explored the ammonia nitrogen removal process of strain K17 on simulated leaching liquor of the rare earth element leaching site. Based on the determination of ammonia nitrogen removal and enzyme activity, it was found that strain K17 has both heterotrophic nitrifying and aerobic denitrifying activities. In addition, single-factor experiments revealed that the most appropriate carbon source for strain K17 was sodium citrate with a C/N ratio of 10 and an initial NH4 +-N concentration of 100 mg/l. Furthermore, the optimal initial pH and rotation speed were 7 and 165 r/min, respectively. Under optimal conditions, the ammonia nitrogen removal efficiency of strain K17 was greater than 95%. As an indigenous bacterium, strain K17 has great potential for treating residual ammonium leaching solutions from rare earth element leaching sites.

Recovery of anammox process performance after substrate inhibition: Reactor performance, sludge morphology, and microbial community.

Most of the current studies have focused on the inhibition of anaerobic ammonium oxidation (anammox) by substrates, however, little attention has been paid to the recovery process of the reactor after inhibition. Therefore, we investigated the changes in reactor performance, granular sludge structure, and microbial community during the recovery phase after being inhibited by a high nitrogen load for 15 d. The nitrogen removal rate of the reactorwasrestored to pre-inhibition levels after 75 d of recovery, and the stoichiometric ratio converged to the theoretical value. The surface of the granular sludge developed into a broccoli-like structure, and the Ca and P contents of the granules increased from 6.88% and 4.39% to 24.42% and 13.88%, respectively. The abundance of the anammox bacterium Candidatus brocadia increased from 5.86% to 12.10%, and network analysis indicated that SMA102 and SBR1031 were positively correlated with the occurrence of Candidatus brocadia.

ECO: An Integrated Gene Expression Omnibus for Mouse Endothelial Cells In Vivo.

Endothelial cell (EC) plays critical roles in vascular physiological and pathological processes. With the development of high-throughput technologies, transcriptomics analysis of EC has increased dramatically and a large amount of informative data have been generated. The dynamic patterns of gene expression in ECs under various conditions were revealed. Unfortunately, due to the lack of bioinformatics infrastructures, reuse of these large-scale datasets is challenging for many scientists. Here, by systematic re-analyzing, integrating, and standardizing of 203 RNA sequencing samples from freshly isolated mouse ECs under 71 conditions, we constructed an integrated mouse EC gene expression omnibus (ECO). The ECO database enables one-click retrieval of endothelial expression profiles from different organs under different conditions including disease models, genetic modifications, and clinically relevant treatments in vivo. The EC expression profiles are visualized with user-friendly bar-plots. It also provides a convenient search tool for co-expressed genes. ECO facilitates endothelial research with an integrated tool and resource for transcriptome analysis. The ECO database is freely available at https://heomics.shinyapps.io/ecodb/.

Targeting Lipid Rafts as a Rapid Screening Strategy for Potential Antiadipogenic Polyphenols along with the Structure-Activity Relationship and Mechanism Elucidation.

Obesity is a global public health problem that endangers human health, and a rapid search for compounds with antiadipogenic activity could provide solutions to overcome this problem. Polyphenols are potential antiadipogenic compounds, but the screening strategy, structure-activity relationship (SAR), and elucidation of their mechanisms of action remain poorly understood because of the high diversity of polyphenols. Lipid rafts, enriched with sphingolipids and cholesterol, are considered a potential target of polyphenols for the regulation of cellular processes and diseases. Here, a novel rapid screening active polyphenol strategy that targets the lipid rafts using molecular dynamic simulation was developed and validated by 3T3-L1 preadipocyte assay. The screening strategy is high-throughput, inexpensive, reagent-free, and effort saving. In addition, the SAR and mechanisms of action mediating the differentiation-inhibition of the preadipocyte by polyphenols were well elucidated by utilizing multiple technologies, such as "raft-like liposomes" systems, giant plasma membrane vesicles, noninvasive lipid raft probes, and ultrahigh-resolution microscopy. High inhibitory-activity polyphenols could penetrate deeper into the hydrophobic lipid center, in an inverted V-shaped manner or by insertion of galloyl groups into rafts, thus disrupting the ordered domain of lipid rafts. In contrast, the medium and low inhibitory-activity polyphenols could only localize on the surface of lipid rafts, exerting slight and the weakest interference with a lipid raft structure, respectively. The combined use of reliable technologies could yield new knowledge on the SAR and the molecular mechanisms of polyphenols.

Short-term waterlogging-induced autophagy in root cells of wheat can inhibit programmed cell death.

Autophagy is a pathway for the degradation of cytoplasmic components in eukaryotes. In wheat, the mechanism by which autophagy regulates programmed cell death (PCD) is unknown. Here, we demonstrated that short-term waterlogging-induced autophagy inhibited PCD in root cells of wheat. The waterlogging-tolerant wheat cultivar Huamai 8 and the waterlogging-sensitive wheat cultivar Huamai 9 were used as experimental materials, and their roots were waterlogged for 0-48 h. Waterlogging stress increased the number of autophagic structures, the expression levels of autophagy-related genes (TaATG), and the occurrence of PCD in root cells. PCD manifested as morphological changes in the cell nucleus, significant enhancement of DNA laddering bands, and increases in caspase-like protease activity and the expression levels of metacaspase genes. The autophagy promoter rapamycin (RAPA) reduced PCD levels, whereas the autophagy inhibitor 3-methyladenine (3-MA) enhanced them. The expression levels of TaATG genes and the number of autophagic structures were lower in cortex cells than in stele cells, but the levels of PCD were higher in cortex cells. The number of autophagic structures was greater in Huamai 8 than in Huamai 9, but the levels of PCD were lower. In summary, our results showed that short-term waterlogging induced autophagy which could inhibit PCD. Mechanisms of response to waterlogging stress differed between cortex and stele cells and between two wheat cultivars of contrasting waterlogging tolerance.

Mutual regulation of ROS accumulation and cell autophagy in wheat roots under hypoxia stress.

Here, we explored the mutual regulation of radical oxygen species (ROS) and autophagy in wheat (Triticum aestivum L.) roots under hypoxia stress. We also analyzed differences between the responses of the stele and the cortex in the two wheat cultivars Huamai 8 (waterlogging-tolerant) and Huamai 9 (waterlogging-sensitive) to hypoxia stress. In situ detection and ultracytochemical localization analysis in wheat roots showed that hypoxia stress caused greater increases in ROS levels and the expression levels of alternative oxidase (AOX) and antioxidant enzymes in the stele than in the cortex. The analysis of exogenous ROS addition and the inhibition of its production revealed the pivotal roles played by ROS in autophagy. Moreover, transmission electron microscopy and qRT-PCR analysis indicated that the stele had a higher level of autophagy than the cortex and that the two wheat cultivars primarily differed in the type and number of autophagosomes. Additional research revealed that autophagy could remove excess ROS, as pre-treatment with the autophagy inhibitor 3-methyladenine increased ROS levels in roots and the addition of the autophagy inducer rapamycin reduced root ROS levels. In conclusion, hypoxia stress induced ROS accumulation in wheat roots where ROS acted as an autophagy signal. Furthermore, higher levels of autophagy and antioxidant enzyme expression in the stele facilitated the elimination of oxidative damage caused by excessive ROS and thereby increased cell survival; in the cortex, a large number of cells died and formed aerenchyma.

Use of a locking intramedullary nail for the treatment of initial varus proximal humeral fracture: a prospective pilot study.

OBJECTIVE: To evaluate the feasibility of locked intramedullary nailing, rather than locking plate fixation combined with fibular allograft augmentation, for initial varus proximal humeral fractures. METHODS: This prospective pilot study enrolled patients with initial varus proximal humeral fractures that were treated with a locking intramedullary nail. Radiography was performed to evaluate fracture healing. Data about the visual analogue scale (VAS) pain score, Constant Shoulder Score (CSS), Disabilities of the Arm, Shoulder and Hand (DASH) score, American Shoulder and Elbow Surgeons (ASES) score and shoulder range of motion (ROM) were recorded. RESULTS: Twenty patients, including eight with Neer two-part and 12 with three-part fractures, were followed-up, with a mean time of 12.3 months. All patients sustained fractures that healed without re-varus. During the last follow-up, the shoulder function of the patients had recovered well, with a mean VAS pain score of 1.4, a mean CSS of 83.1, a mean DASH score of 80.8, a mean ASES score of 84.0 and a satisfactory ROM. In one patient, the proximal locking screw came out and was removed via a second surgery. CONCLUSIONS: The use of a locking intramedullary nail alone for initial varus proximal humeral two-/three-part fractures was feasible. This treatment has advantages, such as preventing re-varus and causing milder surgical trauma, than that seen with a locking plate.

An Integrated Transcriptome Analysis Reveals IGFBP7 Upregulation in Vasculature in Traumatic Brain Injury.

Vasculature plays critical roles in the pathogenesis and neurological repair of traumatic brain injury (TBI). However, how vascular endothelial cells respond to TBI at the molecular level has not been systematically reviewed. Here, by integrating three transcriptome datasets including whole cortex of mouse brain, FACS-sorted mouse brain endothelial cells, and single cell sequencing of mouse brain hippocampus, we revealed the key molecular alteration of endothelial cells characterized by increased Myc targets and Epithelial-Mesenchymal Transition signatures. In addition, immunofluorescence staining of patients' samples confirmed that IGFBP7 was up-regulated in vasculature in response to TBI. TGFß1, mainly derived from microglia and endothelial cells, sufficiently induces IGFBP7 expression in cultured endothelial cells, and is significantly upregulated in response to TBI. Our results identified IGFBP7 as a potential biomarker of vasculature in response to TBI, and indicate that TGFß signaling may contribute to the upregulation of IGFBP7 in the vasculature.

Effect of Waterlogging-Induced Autophagy on Programmed Cell Death in Arabidopsis Roots.

Autophagy, a highly conserved process in eukaryotes that involves vacuolar degradation of intracellular components and decomposition of damaged or toxic constituents, is induced by endogenous reactive oxygen species (ROS) accumulation, endoplasmic reticulum stress, and other factors. In plants, the role of autophagy in the induction of programmed cell death (PCD) is still unclear. Here, we show that ROS contribute to the regulation of PCD during waterlogging (which results in oxygen depletion) via autophagy. In wild-type roots, waterlogging induces the transcription of hypoxia-responsive genes and respiratory burst oxidase homolog (RBOH)-mediated ROS production. It also altered the transcription level of alternative oxidase1a and the activity level of antioxidant enzymes. Moreover, waterlogging increased the transcription levels of autophagy-related (ATG) genes and the number of autophagosomes. Autophagy first occurred in the root stele, and then autophagosomes appeared at other locations in the root. In rboh mutants, upregulation of autophagosomes was less pronounced than in the wild type upon waterlogging. However, the accumulation of ROS and the level of cell death in the roots of atg mutants were higher than those in the wild type after waterlogging. In conclusion, our results suggest that autophagy induced in Arabidopsis roots during waterlogging has an attenuating effect on PCD in the roots.

The release of cytochrome c and the regulation of the programmed cell death progress in the endosperm of winter wheat (Triticum aestivum L.) under waterlogging.

It has been shown in mammalian systems that the mitochondria can play a key role in the regulation of apoptosis by releasing intermembrane proteins (such as cytochrome c) into the cytosol. Cytochrome c released from the mitochondria to the cytoplasm activates proteolytic enzyme cascades, leading to specific nuclear DNA degradation and cell death. This pathway is considered to be one of the important regulatory mechanisms of apoptosis. Previous studies have shown that endosperm cell development in wheat undergoes specialized programmed cell death (PCD) and that waterlogging stress accelerates the PCD process; however, little is known regarding the associated molecular mechanism. In this study, changes in mitochondrial structure, the release of cytochrome c, and gene expression were studied in the endosperm cells of the wheat (Triticum aestivum L.) cultivar "huamai 8" during PCD under different waterlogging durations. The results showed that waterlogging aggravated the degradation of mitochondrial structure, increased the mitochondrial permeability transition (MPT), and decreased mitochondrial transmembrane potential (ΔΨm), resulting in the advancement of the endosperm PCD process. In situ localization and western blotting of cytochrome c indicated that with the development of the endosperm cell, cytochrome c was gradually released from the mitochondria to the cytoplasm, and waterlogging stress led to an advancement and increase in the release of cytochrome c. In addition, waterlogging stress resulted in the increased expression of the voltage-dependent anion channel (VDAC) and adenine nucleotide translocator (ANT), suggesting that the mitochondrial permeability transition pore (MPTP) may be involved in endosperm PCD under waterlogging stress. The MPTP inhibitor cyclosporine A effectively suppressed cell death and cytochrome c release during wheat endosperm PCD. Our results indicate that the mitochondria play important roles in the PCD of endosperm cells and that the increase in mitochondrial damage and corresponding release of cytochrome c may be one of the major causes of endosperm PCD advancement under waterlogging.

Persimmon Tannin Decreased the Glycemic Response through Decreasing the Digestibility of Starch and Inhibiting α-Amylase, α-Glucosidase, and Intestinal Glucose Uptake.

Regulation of postprandial blood glucose levels is an effective therapeutic proposal for type 2 diabetes treatment. In this study, the effect of persimmon tannin on starch digestion with different amylose levels was investigated both in vitro and in vivo. Oral administration of persimmon tannin-starch complexes significantly suppressed the increase of blood glucose levels and the area under the curve (AUC) in a dose-dependent manner compared with starch treatment alone in an in vivo rat model. Further study proved that persimmon tannin could not only interact with starch directly but also inhibit α-amylase and α-glucosidase strongly, with IC50 values of 0.35 and 0.24 mg/mL, separately. In addition, 20 µg/mL of persimmon tannin significantly decreased glucose uptake and transport in Caco-2 cells model. Overall, our data suggested that persimmon tannin may alleviate postprandial hyperglycemia through limiting the digestion of starch as well as inhibiting the uptake and transport of glucose.

Dihydrofisetin exerts its anti-inflammatory effects associated with suppressing ERK/p38 MAPK and Heme Oxygenase-1 activation in lipopolysaccharide-stimulated RAW 264.7 macrophages and carrageenan-induced mice paw edema.

Dihydrofisetin is a flavanonol derived from some edible wild herbs and traditional Chinese medicines. It has been found to possess many biological activities. However, the anti-inflammatory potential of Dihydrofisetin remains uncharacterized. The aim of the present study was to investigate the anti-inflammatory activity of Dihydrofisetin and its underlying mechanisms. We found that Dihydrofisetin dose-dependently inhibited lipopolysaccharide-induced productions of nitric oxide (NO) and prostaglandin E2 (PGE2) in RAW 264.7 macrophages, probably through suppressing the protein expressions of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). The expressions of pro-inflammatory cytokines, such as tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß), interleukin-6 (IL-6) and monocyte chemotactic protein (MCP-1) were also suppressed. We further demonstrated that Dihydrofisetin inhibited the activation of mitogen-activated protein kinases (MAPKs) pathway and phosphorylation of IκB-α whereas upregulated the expression of heme oxygenase-1 (HO-1). The in vivo carrageenan-induced mice paw edema study also indicated that treatment with 100 mg/kg of Dihydrofisetin could significantly inhibit carrageenan induced paw edema, decrease the levels of TNF-α, IL-6 and MDA, and increase the activity of GSH-Px in paw tissues. Taken together, Dihydrofisetin may act as a natural agent for treating inflammatory diseases by targeting MAPK, NF-κB and HO-1 pathways.

Reshaped fecal gut microbiota composition by the intake of high molecular weight persimmon tannin in normal and high-cholesterol diet-fed rats.

It has been proposed that the gut microbiome may be related to obesity, and diet-induced obesity may induce changes in the gut microbiota composition. Our previous studies suggested that persimmon tannin (PT), which is highly polymerized and non-absorbable in the intestine, showed anti-hyperlipidemic and cholesterol-lowering effects in animal models. Considering that the possible composition modification effects of PT on intestinal bacteria might contribute to its anti-hyperlipidemic and cholesterol-lowering effects in vivo, in this study, we determined whether the PT administration could modify the gut microbiota in both normal diet-fed and high-cholesterol (HC) diet-fed rats, and how PT altered the bacterial composition in both normal and HC fed rats. Sprague-Dawley rats were randomly divided into eight groups, and fed with either a normal or an HC diet supplemented with or without a low/medium/high dose of PT (50 (LPT), 100 (MPT), 200 (HPT) mg per kg body weight (BW) per day, respectively) for 4 weeks. On days 0, 7, 14, 21 and 28, feces were collected and prepared for the microbiota and physicochemical analysis. The results showed that LPT and MPT supplementation significantly altered the gut microbiota composition by increasing the Bacteroidetes/Proteobacteria ratio in both normal diet-fed and HC diet-fed rats. LPT also decreased the Firmicutes/Bacteroidetes ratio in normal diet-fed rats and MPT decreased the Firmicutes/Bacteroidetes ratio in HC diet-fed rats. Both LPT and MPT supplementation induced a significant alteration in specific bacterial species after 14 days of treatment. The relative abundance of Bifidobacterium sp. and Lactobacillus sp. was increased by both LPT and MPT treatment, and that of Escherichia coli and Enterococcus was reduced. Our data also indicate that there is a correlation between the changes in bacterial composition and the changes in short-chain fatty acid (SCFA) metabolism. However, HPT supplementation altered the gut microbiota at the phylum and species levels in an adverse way.

Separation and purification of four phenolic compounds from persimmon by high-speed counter-current chromatography.

An efficient method was established by high-speed counter-current chromatography (HSCCC) for preparation of four phenolic compounds from the depolymerization products of persimmon tannin. Using the two solvent systems of n-hexane/ethyl acetate/water (3:17:20, v/v/v) and ethyl acetate/methanol/water (50:1:50, v/v/v), the preparative isolation was successfully performed by a two-step separation. The yields of one run (150mg crude sample) for gallic acid, methyl gallate, and epigallocatechin-3-gallate-(4ß→8, 2ß→O→7)-epigallocatechin-3-gallate dimer (A-type EGCG dimer) were 4.7, 44.2 and 5.9mg, respectively. In addition, 4.6mg epicatechin-3-gallate-(4ß→8, 2ß→O→7)-epicatechin-3-gallate dimer (A-type ECG dimer) was obtained by further preparative high-performance liquid chromatography (prep-HPLC). The purities of these compounds were all above 95.0% and their structures were identified by HPLC/ESI-MS. We found that HSCCC had definite advantages for the preparation of dimeric procyanidins compared with previous methods. Furthermore, it was shown that the four phenolic compounds possessed greater antioxidant activities than Trolox.