Penthorum chinense Pursh leaf tea debittering mechanisms via green tea manufacturing process and its influence on NAFLD-alleviation activities.

The green-tea manufacturing process showed good effect of flavor improving, debittering and shaping in making Penthorum chinensePursh leaf (PL) tea (PLT), which serves as a polyphenol dietary supplement and beverage raw material. GC-MS results showed that its unpleasant grassy odor decreased by 42.8% due to dodecanal, geranylacetone, and (E)-2-nonenal reduction, coupled with 1-hexadecanol increasing. UPLC-ESI-TOF-MS identified 95 compounds and showed that the debittering effect of green-tea manufacturing process was attributed to decreasing of flavonols and lignans, especially quercetins, kaempferols and luteolins, and increasing of dihydrochalcones which act as sweeteners bitterness-masking agents, while astringency was weakened by reducing delphinidin-3,5-O-diglucoside chloride, kaempferol-7-O-ß-d-glucopyranoside, and tannins. The increase of pinocembrins and catechins in aqueous extracts of PLT, maintained its hepatoprotective, NAFLD-alleviation, and hepatofibrosis-prevention activities similar to PL in high fat-diet C57BL/6 mice, with flavonoids, tannins, tannic acids, and some newfound chemicals, including norbergenin, gomisin K2, pseudolaric acid B, tanshinol B, as functional ingredients.

Comparative Analysis of Volatile Flavor Compounds in Strongly Flavored Baijiu under Two Different Pit Cap Sealing Processes.

The solid-state fermentation process of strongly flavored Baijiu is complicated by the co-fermentation of many different microorganisms in the fermentation pools. The traditional fermentation pools of strong flavor Baijiu are sealed with mud, and this sealed-pit mud is not easy to maintain; therefore, the pit cap is prone to cracks and to caving in. The destruction of the sealed-pit mud may lead to instability in the composition and an abundance of microorganisms in the fermentation process that results in fluctuations of product quality. Thus, the production method of replacing the mud cap with a new steel cap is gradually attracting the attention of scientific and technical workers in the industry. However, so far, there have been relatively few reports on the use of steel lids for sealing pits for fermentation and brewing. In this study, the volatile flavor components of 270 Baijiu samples from mud-sealing and steel-sealing pits of a Chinese Baijiu distillery were studied qualitatively and quantitatively using Gas Chromatography-Mass Spectrometry (Abbreviated as GC-MS). Our statistical methods included Hierarchical Cluster Analysis (Abbreviated as HCA), Principal Component Analysis (Abbreviated as PCA), and Discriminant Analysis (Abbreviated as DA). A statistical analysis was carried out on the yield of strongly flavored Baijiu, and we made a comprehensive evaluation of the Baijiu produced under the two pit-sealing modes with regard to flavor and economic efficiency. The yield of strong flavored Baijiu was 6.7% higher with steel-sealing pits compared with mud-sealing pits. Cluster analysis categorized the strongly flavored Baijiu samples into two categories initially: (1) samples produced using mud-sealing pits and (2) samples using steel-sealing pits. Our analysis also indicated that the 28 compounds used for quantification were selected correctly. Surprising to the experimental staff, the overall score for the steel-sealing pits was greater than that of the mud-sealing pits based on PCA. Using DA, the prediction results were 100% accurate. In summary, through a comparative analysis of the flavor and yield, which are the two main factors that affect the quality of Baijiu in a distillery, and systematic combination at both experimental and theoretical levels, the differences between the Baijiu production by steel-sealing and the traditional mud-sealing were clear. Regardless of the impact of age, the detectable flavor components of Baijiu from the mud-steeling pits were very consistent with those of the steel-sealing pits in terms of richness or concentration. However, steel-sealing pits were significantly superior to mud-sealing pits with respect to output, consistency in quality, and cost (human and economic) savings.

Preventive effect and mechanism of Tibetan tea extract on thrombosis in arachidonic acid-induced zebrafish determined via RNA-seq transcriptome profiles.

Thrombosis is a key pathological event in cardiovascular diseases and is also the most important targeting process for their clinical management. In this study, arachidonic acid (AA) was used to induce thrombus formation in zebrafish larvae. Blood flow, red blood cell (RBCs) aggregation and cellular oxidative stress were measured to evaluate the antithrombotic effect of Tibetan tea (TT). Meanwhile, the potential molecular mechanism was further explored by transcriptome sequencing (RNA-seq). The results indicated that TT could significantly restore heart RBCs intensity of thrombotic zebrafish, whilst decreasing RBCs accumulation in the caudal vein. The transcriptome analysis revealed that the preventive effect of TT on thrombosis could be mostly attributed to changes in lipid metabolism related signaling pathways, such as fatty acid metabolism, glycerollipid metabolism, ECM-receptor interaction and steroid biosynthesis signaling pathway. This study demonstrated that Tibetan tea could alleviate thrombosis by reducing oxidative stress levels and regulating lipid metabolism.

Metabolomic Analysis of Fermented Tibetan Tea Using Bacillus circulans and Their Biological Activity on Mice via the Intestine-Hepatic Axis.

The use of Bacillus circulans as the sole starter provides better process control compared to natural fermentation. However, the chemical composition of fermented Tibetan tea by B. circulans and its regulatory effects on the intestine-liver axis has not been reported. For this purpose, a high-resolution liquid chromatography tandem mass spectrometry metabolomics approach was performed. The effects of fermented Tibetan tea on the intestine-liver axis of mice were also evaluated. Untargeted metabolomics analysis showed that the contents of catechin derivatives, flavonoids, phenolic acids, and terpenoids increased by 0.3, 2.38, 2.65, and 3.36%, respectively, compared with those before fermentation. Furthermore, 16S ribosomal RNA sequence analysis revealed that the relative abundance of Lactobacillus spp. in the intestine increased after consumption of fermented tea. Additionally, based on histological and quantitative PCR analyses, fermented Tibetan tea also improved intestinal development and intestinal barrier function in mouse, while increasing the antioxidant capacity of mouse liver. Thus, fermented Tibetan tea could provide beneficial health effects through the intestine-liver axis. These findings have facilitated the study of the chemical composition of Tibetan tea and provided theoretical support for its use as a natural beverage with intestinal probiotic functions.

Major Histocompatibility Complex (MHC) Diversity of the Reintroduction Populations of Endangered Przewalski's Horse.

Major histocompatibility complex (MHC) genes are the most polymorphic in vertebrates and the high variability in many MHC genes is thought to play a crucial role in pathogen recognition. The MHC class II locus DQA polymorphism was analyzed in the endangered Przewalski's horse, Equus przewalskii, a species that has been extinct in the wild and all the current living individuals descend from 12 founders. We used the polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) to detect the polymorphism within the MHC DQA in 31 Przewalski's horses from two reintroduced populations. Consequently, only seven alleles were identified, with only four presenting in each population. In comparison with other mammals, the Przewalski's horse demonstrated less MHC variation. The nucleotide genetic distance of the seven ELA-DQA alleles was between 0.012 and 0.161. The Poisson corrected amino acid genetic distance of the founded alleles was 0.01-0.334. The allele and genotype frequencies of both reintroduced populations of Przewalski's horse deviated from the Hardy-Weinberg equilibrium. Specific MHC DQA alleles may have been lost during the extreme bottleneck event that this species underwent throughout history. We suggest the necessity to detect the genetic background of individuals prior to performing the reintroduction project.

Differentiation and classification of Chinese Luzhou-flavor liquors with different geographical origins based on fingerprint and chemometric analysis.

In order to differentiate and characterize Chinese Luzhou-flavor liquor according to geographical origins, the volatile flavor compounds were analyzed for forty commercial Luzhou-flavor liquor samples from Sichuan, Jiangsu, and Hubei provinces. A total of 113 volatile flavor compounds were quantified; among them, 29 flavor compounds were quantified according to the internal standard method. The differences in flavor composition among different brands of Luzhou-flavor liquor were compared. A data matrix of 64 (flavor components) × 40 (samples) was studied and interpreted using chemometric analysis. The research object could be naturally clustered according to geographical origin (brand) based on the hierarchical cluster analysis (HCA), principal component analysis (PCA) and multivariate analysis of variance (MANOVA) methods. A 100% of predication ability was obtained by the application of K-nearest neighbor model (KNN) for study sample classification. The results demonstrate that the abundance of volatile flavor components in liquors combined with appropriate multivariate statistical methods could be used for the division and traceability of liquors from different geographic origins. PRACTICAL APPLICATION: This study can provide the basis for the identification of liquor authenticity and the traceability of liquor.

Relationship between Brain Natriuretic Peptide and Recurrence of Atrial Fibrillation after Successful Electrical Cardioversion: an Updated Meta-Analysis.

OBJECTIVE: To investigate the relationship between brain natriuretic peptide and recurrence of atrial fibrillation after successful electrical cardioversion. METHODS: Medline and Embase databases were used to identify publications evaluating BNP/N-Terminal (NT)-proBNP levels in association with atrial fibrillation recurrence after successful electrical cardioversion. Nineteen studies that fulfilled the specified criteria of our analysis were found. RESULTS: Baseline BNP/NT-proBNP levels of the atrial fibrillation recurrence group were significantly higher than those of the sinus rhythm maintaining group (SMD -0.70, CI [-0.82, -0.58]). CONCLUSION: Our analysis suggests that low BNP/NT-proBNP levels are associated with sinus rhythm maintenance, and baseline BNP/NT-proBNP concentrations may be a predictor of atrial fibrillation recurrence after successful electrical cardioversion.

Relationship between brain natriuretic peptide and recurrence of atrial fibrillation after successful electrical cardioversion: an updated meta-analys

Abstract Objective: To investigate the relationship between brain natriuretic peptide and recurrence of atrial fibrillation after successful electrical cardioversion. Methods: Medline and Embase databases were used to identify publications evaluating BNP/N-Terminal (NT)-proBNP levels in association with atrial fibrillation recurrence after successful electrical cardioversion. Nineteen studies that fulfilled the specified criteria of our analysis were found. Results: Baseline BNP/NT-proBNP levels of the atrial fibrillation recurrence group were significantly higher than those of the sinus rhythm maintaining group (SMD -0.70, CI [-0.82, -0.58]). Conclusion: Our analysis suggests that low BNP/NT-proBNP levels are associated with sinus rhythm maintenance, and baseline BNP/NT-proBNP concentrations may be a predictor of atrial fibrillation recurrence after successful electrical cardioversion.

Association between a vascular endothelial growth factor gene polymorphism (rs2146323) and diabetic retinopathy: a meta-analysis.

BACKGROUND: Vascular endothelial growth factor (VEGF) is thought to play an important role in the pathogenesis of diabetic retinopathy (DR). Previous studies have associated the VEGF rs2146323 polymorphism with the risk of DR. However, the results of these studies are inconsistent. A meta-analysis was performed to evaluate the association between the VEGF rs2146323 polymorphism and the risk of DR. METHODS: The PubMed, EMBASE, Web of Science and Google Scholar literature databases until March 2015 were searched. The differences in the studies were expressed in the form of an odds ratio (OR) and the corresponding 95% confidence interval (CI). Heterogeneity among the studies was tested using the I(2) statistic based on the Q test. RESULTS: A total of four studies (598 cases and 709 controls) were included in the meta-analysis. A significant association was found involving the rs2146323 polymorphism in the dominant model (CA + AA VS. CC) (OR = 1.38, CI = 1.10-1.72, P = 0.005) and the co-dominant model (CA VS. CC) (OR = 1.37, CI = 1.08-1.74, P = 0.008). CONCLUSIONS: Our meta-analysis confirmed the association between the VEGF rs2146323 polymorphism and the risk of DR.

Bisoprolol reverses epinephrine-mediated inhibition of cell emigration through increases in the expression of ß-arrestin 2 and CCR7 and PI3K phosphorylation, in dendritic cells loaded with cholesterol.

The effect of bisoprolol on dendritic cell (DC) migration was investigated, including the analysis of protein expression, cytokine secretion and activation of the PI3K-pathway. The chemotactic cell numbers in cholesterol-loaded DCs treated with epinephrine were significantly decreased by 26.66±6.29% (6h), 35.67±2.91% (12h) and 29.33±1.09% (24h). This effect was significantly reversed by 46.00±10.65% (6h), 64.25±6.77% (12h) and 55.74±5.51% (24h) when bisoprolol and epinephrine were both present. In cholesterol-loaded DCs, treatment with epinephrine significantly increased AR-ß1 protein expression by 56.99±4.87%, but inhibited ß-arrestin 2 and CCR7 protein expression by 30.51±4.22% and 25.31±0.04%, respectively. These effects were reversed by bisoprolol by 36.87±4.40%, 41.47±3.95% and 30.14±0.54%, respectively. TNF-α and MMP9 levels were decreased by 68.33±4.00% and 39.57±9.21% in cholesterol-loaded DCs treated with epinephrine. In contrast, when bisoprolol and epinephrine were administered together, the secretion of these proteins was significantly increased by 233.81±37.06 % and 76.66±14.21%, respectively. Treatment with epinephrine decreased PI3K-phosphorylation by 31.88±2.79%, 40.24±5.69% and 30.93±4.66% at 15, 30 and 60min, respectively, whereas the effect of epinephrine on the expression of phosphorylated PI3K was reversed by 49.49±12.12%, 70.93±16.14% and 47.62±6.00%, respectively, when cells were treated with both bisoprolol and epinephrine. Wortmannin inhibited the effects of bisoprolol on PI3K-phosphorylation (38.63±6.12%), the expression of CCR7 (23.4±2.72%), the secretion of TNF-α (69.46±4.48%) and MMP9 (43.15±4.63%), and the number of chemotactic cells (36.84±5.22%). This is the first study to establish a signaling pathway, epinephrine-AR-ß1-ß-arrestin2-PI3K-MMP9/CCR7, which plays a critical role in the migration of DCs.

Effect of overexpression of human SR-AI on oxLDL uptake and apoptosis in 293T cells.

Type I class A macrophage scavenger receptor (SR)-AI plays an important role in foam cell formation and in apoptosis in atherosclerosis, however the mechanism remains unclear. Therefore, we generated a pEGFP-C1-SR-AI plasmid construct for transient transfection of 293T human embryonic kidney cells and observed if SR-AI expression led: (i) to foam cell formation or apoptosis; and (ii) to expression of apoptosis-related genes Bcl-2 and Bak-1 in cells treated with oxidized low-density lipoprotein (oxLDL). The pEGFP-C1 (empty vector) transfected cell line was used as a control. Transfection efficiency of each group was >90% and transfected cells expressed functional SR-AI protein. Binding and uptake of 3,3'-dioctadecylindocarbocyanine-labeled oxLDL (DiI-oxLDL) were verified by flow cytometry; increases in the rate of oxLDL binding and uptake were observed in pEGFP-C1-SR-AI transfected 293T cells and incubation with oxLDL also led to increased apoptosis (≈50%) compared with controls. A decrease in Bcl-2 and an increase in Bak-1 mRNA and protein expression were observed in pEGFP-C1-SR-AI transfected cells compared with controls. We conclude that transient over-expression of SR-AI leads to an increase in oxLDL uptake and binding in a non-macrophage cell line. In addition, over-expression of SR-AI induced non-macrophage cell apoptosis via downregulation of Bcl-2 and upregulation of Bak-1 expression. We conclude that the 293T cell expression described here is a model for foam cell formation. These results may form the basis of further research into SR-AI structure and function (including lipoprotein uptake, apoptosis modulation and adhesion), which may give an insight into the progression of atherosclerosis in vivo.

Interleukin-10 down-regulates oxLDL induced expression of scavenger receptor A and Bak-1 in macrophages derived from THP-1 cells.

Here, we investigated the therapeutic potential of IL-10 by testing its effects on oxLDL-induced lipoprotein uptake and apoptosis by flow cytometry in THP-1-derived macrophages. The mRNA and protein expressions of lipid scavenger receptors (SR-A, CD36) and apoptosis-related proteins (Bcl-2, Bak-1) were also detected. Co-incubation of oxLDL with IL-10 reduced DiI-oxLDL uptake by 16.1±3.8%, 35.2±3.8% and 28.9±1.8% at 6, 12 and 24h of treatment, respectively. Furthermore, treatment with oxLDL for 24h enhanced the SR-A mRNA and protein expressions by 89.3±17.1% and 70.1±17.6%, respectively. IL-10 abrogated the oxLDL-induced SR-A mRNA expression by 50.2±3.9% and its protein by 45.6±1.9%. Meanwhile IL-10 had no effect on the oxLDL-induced increase of CD36 expression. IL-10 inhibited the oxLDL-induced cell apoptosis in a time-dependent manner by 17.3±3.3%, 36.4±2.8% and 31.0±4.3% at 6, 12 and 24h, respectively. OxLDL increased Bak-1 mRNA and protein expressions by 38.4±13.3% and 36.9±12.1%, respectively. However co-stimulation of oxLDL with IL-10 for 24h inhibited Bak-1 expression to 28.4±7.2% (mRNA) and 25.7±6.3% (protein). Meanwhile, IL-10 had no effect on the oxLDL-induced decrease of Bcl-2 expression. Our findings suggested that IL-10 reduced the oxLDL-induced lipoprotein uptake and apoptosis partly via down-regulating the oxLDL-induced expression of SR-A and Bak-1 in THP-1-derived macrophages.

[A novel GATA4 mutation leading to congenital ventricular septal defect].

OBJECTIVE: To identify the GATA4 gene mutation of congenital ventricular septal defect (VSD) and study the molecular mechanism of a novel mutation. METHODS: The clinical data and blood samples from 185 unrelated subjects with congenital VSD were collected and evaluated together with 200 healthy individuals. The coding exons and the flanking intron regions of the GATA4 gene were amplified by PCR and sequenced using the di-deoxynucleotide chain termination approach. The GATA4 gene was cloned and the corresponding mutant was acquired by site directed mutagenesis. The recombinant plasmid expressing GATA4 and the reporter vector expressing enhanced green fluorescence protein (EGFP) driven by the promoter of atrial natrium peptide (ANP) gene were transfected into HeLa cells with Lipofectamine. The effect of mutated GATA4 gene on the transcriptional activity of encoded transcriptional factor was analyzed by reverse transcription (RT)-PCR. RESULTS: A novel heterozygous missense GATA4 mutation, c.191G>A was identified in 1 VSD patient. The mutation leads to glycine to glutamic acid change at amino acid residue 64 (G64E) in the GATA4 protein. Functional analysis showed that GATA4 G64E mutation decreased the transcriptional activity of GATA4 transcriptional factor. CONCLUSION: A novel heterozygous missense GATA4 mutation, G64E, was identified in 1 VSD patient. The mutation might cause VSD by impairing the transcriptional activity of GATA4 transcriptional factor.

KCNQ1 loss-of-function mutation impairs gastric acid secretion in mice.

The KCNQ1 channel is abundantly expressed in the gastric parietal cells. Although the functional coupling of KCNQ1 with the H(+)/K(+)-ATPase has already been confirmed on the basis of pharmacological kinetics, the effect of a KCNQ1 loss-of-function mutation on gastric acidification remains unclear. In this study, parietal cells and gastric glands from both C57BL/6 J mice (normal control) and J343 mice (mice with a KCNQ1 loss-of-function mutation) were isolated to study the effects of KCNQ1 on gastric acidification. We found that the mutation limited intracellular acidification of parietal cells and H(+) secretion of the stomach in response to histamine. Thus, a KCNQ1 loss-of-function mutation may impair gastric acid secretion.