Influence of Centrifugation and Transmembrane Treatment on Determination of Polyphenols and Antioxidant Ability for Sea Buckthorn Juice.

When the total phenolic content (TPC) and antioxidant activity of sea buckthorn juice were assayed by spectrophotometry, the reaction solutions were not clarified, so centrifugation or membrane treatment was needed before determination. In order to find a suitable method for determining TPC and antioxidant activity, the effects of centrifugation and nylon membrane treatment on the determination of TPC and antioxidant activity in sea buckthorn juice were studied. TPC was determined by the Folin-Ciocalteau method, and antioxidant activity was determined by DPPH, ABTS, and FRAP assays. For Treatment Method (C): the sample was centrifuged for 10 min at 10,000 rpm and the supernatant was taken for analysis. Method (CF): The sample was centrifuged for 10 min at 4000 rpm, filtered by Nylon 66 filtration membranes with pore size of 0.22 µm, and taken for analysis. Method (F): the sample was filtered by Nylon 66 filtration membranes with pore size of 0.22 µm and taken for analysis. Method (N): after the sample of ultrasonic extract solution reacted completely with the assay system, the reaction solution was filtered by Nylon 66 filtration membranes with pore size of 0.22 µm and colorimetric determination was performed. The results showed that centrifugation or transmembrane treatment could affect the determination of TPC and antioxidant activity of sea buckthorn juice. There was no significant difference (p > 0.05) between methods (CF) and (F), while there was a significant difference (p < 0.05) between methods (C) (F) (N) or (C) (CF) (N). The TPC and antioxidant activity of sea buckthorn juice determined by the four treatment methods showed the same trend with fermentation time, and the TPC and antioxidant activity showed a significant positive correlation (p < 0.05). The highest TPC or antioxidant activity measured by method (N) indicates that method (N) has the least loss of TPC or antioxidant activity, and it is recommended for sample assays.

Autophagy Regulates Whitefly-Symbiont Metabolic Interactions.

Nutritional symbionts are restricted to specialized host cells called bacteriocytes in various insect orders. These symbionts can provide essential nutrients to the host. However, the cellular mechanisms underlying the regulation of these insect-symbiont metabolic associations remain largely unclear. The whitefly Bemisia tabaci MEAM1 hosts "Candidatus Portiera aleyrodidarum" (here, "Ca. Portiera") and "Candidatus Hamiltonella defensa" (here, "Ca. Hamiltonella") bacteria in the same bacteriocyte. In this study, the induction of autophagy by chemical treatment and gene silencing decreased symbiont titers and essential amino acid (EAA) and B vitamin contents. In contrast, the repression of autophagy in bacteriocytes via Atg8 silencing increased symbiont titers, and amino acid and B vitamin contents. Furthermore, dietary supplementation with non-EAAs or B vitamins alleviated autophagy in whitefly bacteriocytes, elevated TOR (target of rapamycin) expression, and increased symbiont titers. TOR silencing restored symbiont titers in whiteflies after dietary supplementation with B vitamins. These data suggest that "Ca. Portiera" and "Ca. Hamiltonella" evade autophagy of the whitefly bacteriocytes by activating the TOR pathway via providing essential nutrients. Taken together, we demonstrate that autophagy plays a critical role in regulating the metabolic interactions between the whitefly and two intracellular symbionts. Therefore, this study reveals that autophagy is an important cellular basis for bacteriocyte evolution and symbiosis persistence in whiteflies. The whitefly symbiosis unravels the interactions between cellular and metabolic functions of bacteriocytes. IMPORTANCE Nutritional symbionts, which are restricted to specialized host cells called bacteriocytes, can provide essential nutrients for many hosts. However, the cellular mechanisms of regulation of animal-symbiont metabolic associations have been largely unexplored. Here, using the whitefly-"Ca. Portiera"/"Ca. Hamiltonella" endosymbiosis, we demonstrate autophagy regulates the symbiont titers and thereby alters the essential amino acid and B vitamin contents. For persistence in the whitefly bacteriocytes, "Ca. Portiera" and "Ca. Hamiltonella" alleviate autophagy by activating the TOR (target of rapamycin) pathway through providing essential nutrients. Therefore, we demonstrate that autophagy plays a critical role in regulating the metabolic interactions between the whitefly and two intracellular symbionts. This study also provides insight into the cellular basis of bacteriocyte evolution and symbiosis persistence in the whitefly. The mechanisms underlying the role of autophagy in whitefly symbiosis could be widespread in many insect nutritional symbioses. These findings provide a new avenue for whitefly control via regulating autophagy in the future.

Dietary supplementation with Clostridium butyricum improves growth performance of broilers by regulating intestinal microbiota and mucosal epithelial cells.

Clostridium butyricum has been widely considered an antibiotic substitute in recent years. It can promote growth performance, improve the immune response and enhance the intestinal barrier function of the host. In the present study, 1-d-old Arbor Acres (AA) broilers were fed C. butyricum (1 × 109 cfu/kg) for 28 d. The transcriptomic characteristics of epithelial cells of the cecal mucosa were determined by RNA-sequence, and the cecal microbiota composition was explored by 16S ribosomal RNA gene sequencing. The changes in the intestinal mucosa of broilers were then analyzed by tissue staining. Gene Ontology (GO) annotations identified substance transport and processes and pathways that might participate in intestinal development and cell viability. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis revealed that the differentially expressed genes are involved in numerous pathways related to amino acid and vitamin metabolism and antioxidant and defensive functions, among others. The relative expression of some genes associated with intestinal barrier function (claudins 2, 15, 19, and 23, tight junction proteins 1, 2, and 3 and mucin 1) was significantly increased in the treatment group (P < 0.05 or P < 0.01). Moreover, the proportion of Firmicutes was higher in the C. butyricum-treated group, whereas the proportion of Proteobacteria was higher in the control group. At the genus level, the relative abundances of Butyricicoccus and Lactobacillus, among other bacteria, were increased after C. butyricum supplementation. The tissue staining analysis showed that the cecal mucosa of broilers was significantly ameliorated after the addition of C. butyricum (P < 0.05 or P < 0.01). These results showed that dietary supplementation with C. butyricum can enhance the antioxidant capacity, mucosal barrier function, and stabilize the cecal microbiota, resulting in improving the growth performance.

Intracellular symbionts drive sex ratio in the whitefly by facilitating fertilization and provisioning of B vitamins.

Symbionts can regulate animal reproduction in multiple ways, but the underlying physiological and biochemical mechanisms remain largely unknown. The presence of multiple lineages of maternally inherited, intracellular symbionts (the primary and secondary symbionts) in terrestrial arthropods is widespread in nature. However, the biological, metabolic, and evolutionary role of co-resident secondary symbionts for hosts is poorly understood. The bacterial symbionts Hamiltonella and Arsenophonus have very high prevalence in two globally important pests, the whiteflies Bemisia tabaci and Trialeurodes vaporariorum, respectively. Both symbionts coexist with the primary symbiont Portiera in the same host cell (bacteriocyte) and are maternally transmitted. We found that elimination of both Hamiltonella and Arsenophonous by antibiotic treatment reduced the percentage of female offspring in whiteflies. Microsatellite genotyping and cytogenetic analysis revealed that symbiont deficiency inhibited fertilization in whiteflies, leading to more haploid males with one maternal allele, which is consistent with distorted sex ratio in whiteflies. Quantification of essential amino acids and B vitamins in whiteflies indicated that symbiont deficiency reduced B vitamin levels, and dietary B vitamin supplementation rescued fitness of whiteflies. This study, for the first time, conclusively demonstrates that these two intracellular symbionts affect sex ratios in their whitefly hosts by regulating fertilization and supplying B vitamins. Our results reveal that both symbionts have the convergent function of regulating reproduction in phylogenetically-distant whitefly species. The 100% frequency, the inability of whiteflies to develop normally without their symbiont, and rescue with B vitamins suggests that both symbionts may be better considered co-primary symbionts.

Alterations of static functional connectivity and dynamic functional connectivity in motor execution regions after stroke.

The aims of this study were to examine both static functional connectivity (FC) and dynamic FC alterations in motor execution regions after stroke and to investigate whether the altered static or dynamic FC was associated with the clinical behaviors in stroke patients. Seventy-six stroke patients and 55 healthy controls (HC) were recruited. Static FC and dynamic FC maps were computed based on the seeds of six core regions in motor execution network. Correlation analyses were performed between static or dynamic FC and clinical behavioral scores in stroke patients. Compared with the HC, the stroke patients had significantly higher static FC between the seeds and the precentral or postcentral gyrus, frontal gyrus, inferior parietal lobule, thalamus and insula, and lower static FC between the seeds and the cerebellum and middle temporal gyrus. There were significant differences in dynamic FC between the seeds and precuneus, calcarine gyrus, insula, inferior parietal lobule, precentral gyrus, and middle temporal, frontal or occipital gyrus between the stroke patients and HC. Furthermore, a significant negative correlation was found between the Fugl-Meyer assessment scores and dynamic FC of the ipsilesional primary motor cortex and contralesional precentral gyrus in patients. The current study shows that the patterns of both static FC and dynamic FC changed after stroke, and correlation between motor function and temporal variability in the FC of the precentral gyrus was significant in stroke patients. Our findings indicate that dynamic FC might be a potential indicator for evaluating motor function after stroke.

Comparative transcriptome analysis of hypothalamus-regulated feed intake induced by exogenous visfatin in chicks.

BACKGROUND: The intracerebroventricular injection of visfatin increases feed intake. However, little is known about the molecular mechanism in chicks. This study was conducted to assess the effect of visfatin on the feeding behavior of chicks and the associated molecular mechanism. RESULTS: In response to the intraventricular injection of 40 ng and 400 ng visfatin, feed intake in chicks was significantly increased, and the concentrations of glucose, insulin, TG, HDL and LDL were significantly altered. Using RNA-seq, we identified DEGs in the chick hypothalamus at 60 min after injection with various doses of visfatin. In total, 325, 85 and 519 DEGs were identified in the treated chick hypothalamus in the LT vs C, HT vs C and LT vs HT comparisons, respectively. The changes in the expression profiles of DEGs, GO functional categories, KEGG pathways, and PPI networks by visfatin-mediated regulation of feed intake were analyzed. The DEGs were grouped into 8 clusters based on their expression patterns via K-mean clustering; there were 14 appetite-related DEGs enriched in the hormone activity GO term. The neuroactive ligand-receptor interaction pathway was the key pathway affected by visfatin. The PPI analysis of DEGs showed that POMC was a hub gene that interacted with the maximum number of nodes and ingestion-related pathways, including POMC, CRH, AgRP, NPY, TRH, VIP, NPYL, CGA and TSHB. CONCLUSION: These common DEGs were enriched in the hormone activity GO term and the neuroactive ligand-receptor interaction pathway. Therefore, visfatin causes hyperphagia via the POMC/CRH and NPY/AgRP signaling pathways. These results provide valuable information about the molecular mechanisms of the regulation of food intake by visfatin.

Structure characterization and biological activities of a pectic polysaccharide from cupule of Castanea henryi.

A pectic polysaccharide (CHIP3) was fractionated from the natural cupule of Castanea henryi. It contained mannose (10.70%), rhamnose (8.70%), galacturonic acid (38.21%), galactose (13.75%) and arabinose (28.63%) with a molecular weight of 2.44 × 104 g/mol by multi-laser light scattering. The structure was elucidated by using FT-IR spectroscopy, methylation analysis and NMR analysis. Results showed that the backbone of CHIP3 consisted of 1, 4-α-linked d-GalpA residues containing the non-methyl-esterified carboxyl groups, interspersed with a few 1,2-α-l-Rhap units. Its side chains were attached by two branches to O-4 of Rhap with 1,4-ß-linked d-Galp units and 1,5-α-l-linked Araf units bearing 3,5-substituted α-l-linked Araf residues as branching points. AFM data revealed it existed as a flexible chain in 0.1 M NaNO3 aqueous solution. Furthermore, CHIP3 was demonstrated to have notable antioxidant activity of FRAP, ABTS+ radical scavenging and reducing power. Cytotoxicity assay showed it displayed inhibitory activity against HepG2 cells with IC50 values of 242.6 µg/mL.

One-pot synthesis of multicolor MnSe:ZnSe nanocrystals for optical coding.

Though the investigation on controlling the fluorescence properties of nanocrystals (NCs) with single emission has been widely reported, few efforts were spent on adjusting the fluorescence properties of NCs with multiple emission peaks. In this work, we successfully synthesized multicolor MnSe:ZnSe NCs with multiple emission peaks and developed a simple and accurate method to realize photoluminescence (PL) spectra (or color) adjustment. The PL of MnSe:ZnSe NCs has two distinct emission peaks, the trap emission of ZnSe at 475nm and Mn(2+)-induced emission at 585nm. By adjusting the nucleation temperature, the emission color of the NCs can be encoded according to the ratio of the emission intensities at 475 and 585nm. With the nucleation temperature rising from 0 to 70°C, the PL ratio between trap emission and Mn(2+)-induced emission can be consecutively changed from (1, 3) to (1, 0.5). In addition, the trap state is deeply inside the NCs rather than on NCs surface so that the trap emission is stable during environment change. Thus, these MnSe:ZnSe NCs hold great promise as novel single-particle coding labels for biomedical imaging.

[Observation and multifactor analysis of refractory medium-severe heart failure by micro-inflammation modification in uremic patients].

OBJECTIVE: To investigate the therapeutic strategies and prognostic factors of refractory medium-severe heart failure in uremic patients. METHODS: A single center, self control clinical research was conducted, and the data consisted of 30 uremic patients with refractory medium-severe heart failure undergoing maintenance hemodialysis (MHD), who received routine combined modality therapy and Xuebijing injection (to modify micro-inflammation). The systolic function of the left ventricle was compared before and after therapy. Multiple linear regression models were established to predict the improvements of systolic function of ventricle. Relationship between the accumulated dose of Xuebijing injection and changes of C-reactive protein (ΔCRP) was observed. RESULTS: The values of left ventricle ejection fraction (LVEF), fractional shortening (FS), and stroke volume (SV) after therapy were improved compared with those before therapy [LVEF: 0.42±0.07 vs. 0.34±0.04, FS: (21.07±3.83)% vs. (16.33±2.43)%, SV: 66.83±7.00 ml vs. 52.20±7.62 ml, all P<0.01]. In terms of cardiac output (CO), there was no statistical difference before and after therapy (4.77±0.65 L/min vs. 4.49±0.68 L/min, P>0.05). In the multiple linear regression models of ΔLVEF, ΔFS and ΔSV, the independent variables that affect dependent variables included age, ΔCRP, changes of hemoglobin (ΔHb), accumulated dose of Xuebijing injection, changes of HCO(3)(-) (Δ HCO(3)(-)), changes of serum creatinine (ΔSCr), Hb and CRP after therapy, the factors and weights of which had slight variation on accordance with different dependent variables. There was significant positive correlation between accumulated dose of Xuebijing injection and ΔCRP (r=0.561, P=0.001). CONCLUSIONS: Xuebijing injection can improve heart function in uremic patients by modifying micro-inflammation, whose accumulated dose and therapeutic effect show positive correlation. In addition the improvement of heart failure has something to do with age, ΔHb, Hb after therapy, the correction of acidosis and dialysis sufficiency.

Polyprenols from Taxus chinensis var. mairei prevent the development of CCl4-induced liver fibrosis in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: The aim of this study was to investigate the anti-fibrotic effects and the possible underlying mechanisms of taxus polyprenols (TPs) isolated from the needles of Taxus chinensis var. mairei. MATERIALS AND METHODS: The animals were randomly divided into normal control with vehicles only (olive oil), rat model given CCl4 only, CCl4+low TPs (48 mg/kg), CCl4+medium TPs (120 mg/kg), CCl4+high TPs (300 mg/kg), and CCl4+Polyene phosphatidylcholine (PP, 120 mg/kg). The rat model of liver fibrosis was induced by subcutaneous injection of 40% (v/v) of CCl4 diluted in olive oil (3 mL/kg body weight) twice per week for 8 weeks. Liver histopathological study was performed. Aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP) and albumin (ALB) of the serum were determined for evaluating the liver function. In order to reveal the possible mechanisms of the anti-fibrotic effects, oxidative stress level, hepatic collagen metabolism, and hepatic stellate cells (HSCs) activation were investigated. Furthermore, the mRNA expression of the fibrotic-related factors was measured by the quantitative real-time RT-PCR. RESULTS: TPs successfully attenuated liver injury induced by CCl4 shown by histopathological sections of livers and improved liver function as indicated by decreased ALT, AST and ALP levels and increased ALB levels in serum of the rats. TPs significantly increased the hepatic Cu/Zn SOD and GSH-Px activities along with GSH content while a remarkable decrease in MDA content. Both immunohistochemical staining and mRNA expression levels of α-SMA indicated a profound suppression of HSCs activation. Furthermore, it significantly inhibited the mRNA expression of the pro-fibrotic cytokines Col α1(I), Col α1(Ш), MMP-2, TIMP-1, TIMP-2, PDGF-ß, TGF-ß1, CTGF and TNF-α and restored the hepatoprotective factor HGF. CONCLUSION: These results suggest that the protective effects of TPs in chronic CCl4-induced liver fibrosis might be related with the reduction of oxidative damage, the inhibition of HSCs activation, the down-regulation of pro-fibrogenic stimuli and the protection of hepatocytes.

Polyprenols from the needles of Taxus chinensis var. mairei.

Polyprenols with various pharmacological activities were first isolated from Taxus chinensis var. mairei, which is native to China, and were identified by a high performance liquid chromatography/mass spectrometry (HPLC/MS), (1)H nuclear magnetic resonance ((1)H NMR), (13)C NMR and infrared (IR) spectroscopy. The taxus polyprenols (TPs) had two maxima of polyprenol-17 and polyprenol-21 and different from polyprenols distributed in other plant species. A reversed-phase HPLC method with a simple gradient elution was established, and the proposed combined peak area method for the quantification of polyprenols was verified. The analysis indicated that TPs were present in the old needles at levels as high as 3% and that T. chinensis var. mairei could be an alternative botanical source for the extraction of polyprenols.