Characteristics of Metabolites in the Development of Atherosclerosis in Tibetan Minipigs Determined Using Untargeted Metabolomics.

BACKGROUND: Atherosclerosis (AS) is a chronic progressive disease caused by lipometabolic disorder. However, the pathological characteristics and mechanism of AS have not been fully clarified. Through high-fat and high-cholesterol diet induction, Tibetan minipigs can be used as the AS model animals, as they have a very similar AS pathogenesis to humans. METHODS: In this study, we built an AS model of Tibetan minipigs and identified the differential abundance metabolites in the development of AS based on untargeted metabolomics. RESULTS: We found that sphingolipid metabolism and glucose oxidation were obviously higher in the AS group and phenylalanine metabolism was reduced in the AS group. Moreover, in the development of AS, gluconolactone was enriched in the late stage of AS whereas biopterin was enriched in the early stage of AS. CONCLUSIONS: Our research provides novel clues to investigate the metabolic mechanism of AS from the perspective of metabolomics.

Guanxinning tablets improve myocardial hypertrophy by inhibiting the activation of MEK-ERK1/2 signaling pathway.

Myocardial hypertrophy may lead to heart failure and sudden death. As traditional Chinese medicine, Guanxinning tablets (GXN) have significant pharmacological effects in the prevention and treatment of cardiovascular diseases. However, the anti-cardiac hypertrophy efficacy of GXN and its mechanism of action are still unclear. Therefore, we established a heart failure rat model and isolated primary cardiomyocytes of neonatal rat to observe the protective effect of GXN on heart failure rat model and the intervention effect on myocardial cell hypertrophy, and to explore the possible mechanism of GXN preventing and treating myocardial hypertrophy. The results of in vivo experiments showed that GXN could significantly reduce the degree of cardiac hypertrophy, reduce the size of cardiomyocytes, inhibit the degree of myocardial remodeling and fibrosis, and improve cardiac function in rats with early heart failure. The results of in vitro experiments showed that GXN was safe for primary cardiomyocytes and could improve cardiomyocyte hypertrophy and reduce the apoptosis of cardiomyocytes in pathological state, which may be related to the inhibition of the over-activation of MEK-ERK1/2 signaling pathway. In conclusion, GXN may inhibit cardiac hypertrophy and improve early heart failure by inhibiting the over-activation of MEK-ERK1/2 signaling pathway.

Guanxinning Tablet Attenuates Coronary Atherosclerosis via Regulating the Gut Microbiota and Their Metabolites in Tibetan Minipigs Induced by a High-Fat Diet.

Coronary atherosclerosis (CA) is a chronic and evolving inflammatory disease characterized by the build-up of atherosclerotic plaque in the wall of coronary arteries. Guanxinning tablet (GXNT) is a novel Chinese medicine formula, which has been clinically used to treat coronary heart disease for many years. However, the potential mechanism for treating CA remains unclear. Thus, the study was aimed at investigating the therapeutic effect of GXNT on CA and further explore the underlying mechanisms from the perspective of gut microbiota. Following the establishment of a CA model in Tibetan minipigs, GXNT was orally administrated. We simultaneously detected blood lipid levels, observed ventricular function using ultrasound examination, measured platelet aggregation, and checked changes in inflammatory factors, oxidative stress factors, and vascular endothelial injury-related indexes applying ELISA assays. Histopathological changes of coronary artery tissue were subsequently evaluated using Sudan IV staining, HE staining, Oil red "O" staining, and immunohistochemistry assays. Finally, alterations of the gut microbiota and microbial metabolites were detected using metagenomic sequencing and targeted metabolomics, respectively. The results have suggested that GXNT could regulate dyslipidemia, improve heart function, and inhibit the levels of ox-LDL, CRP, TNF-α, IL-1ß, SOD, MDA, vWF, and ET-1, as well as platelet aggregation. Additionally, histopathological findings revealed that GXNT could reduce lipid deposition, alleviate AS lesions, and restrain the expressions of NF-κB, TNF-α, and MMP-9. Furthermore, the composition of the gut microbiota was altered. Specifically, GXNT could upregulate the relative abundance of Prevotellaceae and Prevotella and downregulate the abundance of Proteobacteria, Enterobacteriaceae, and Escherichia. As for microbial metabolites, GXNT could increase fecal propionic acid, butyric acid, and LCA-3S and decrease fecal TMA-related metabolites, CDCA, and serum TMAO. In sum, the results showed that GXNT had a satisfactory anti-CA effect, and the mechanism was closely associated with modulating gut microbiota and related metabolites.

Guanxinning tablet inhibits the interaction between leukocyte integrin Mac-1 and platelet GPIbα for antithrombosis without increased bleeding risk.

Recent studies have showed that thrombosis is closely related to leucocytes involved in immunity. Interfering with the binding of leukocyte integrin Mac-1 and platelet GPIbα can inhibit thrombosis without affecting physiological coagulation. Mac-1-GPIbα is proposed as a potential safety target for antithrombotic agents. Guanxinning tablet (GXNT) is an oral Chinese patent medicine used for the treatment of angina pectoris, which contains phenolic acid active ingredients, such as salvianolic acids, ferulic acid, chlorogenic acid, caffeic acid, rosmarinic acid, tanshinol, and protocatechualdehyde. Our previous studies demonstrated that GXN exhibited significant antithrombotic effects, and clinical studies suggested that it did not increase bleeding risk. In addition, GXN exerted a significantly regulatory effect on immune inflammation. In the current study, we intended to evaluate the effects of GXN on bleeding events and explore the safety antithrombotic mechanism of GXN based on leukocyte-platelet interaction. First, we established a gastric ulcer model induced by acetic acid in rats and found that GXN not only did not increase the degree of gastrointestinal bleeding when gastric ulcer occurred, but also had a certain promoting effect on the healing of gastric ulcer. Second, in vitroexperiments showed that after pretreatment with GXN and activation by phorbol 12-myristate-13-acetate (PMA), the adhesion and aggregation of leukocytes with human platelets were reduced. It was also found that GXN reduced the expression and activation of Mac-1 in leucocytes, and inhibited platelet activation due to leukocyte engagement via Mac-1. Overall, the results suggest that GXN may be a safe antithrombotic agent, and its low bleeding risk mechanism is probably related to inhibited leukocyte-platelet aggregation and its interaction target Mac-1-GPIbα.

GuanXinNing Tablet Attenuates Alzheimer's Disease via Improving Gut Microbiota, Host Metabolites, and Neuronal Apoptosis in Rabbits.

Based on accumulating evidence, Alzheimer's disease (AD) is related to hypercholesterolemia, gut microbiota, and host metabolites. GuanXinNing Tablet (GXN) is an oral compound preparation composed of two Chinese herbs, Salvia miltiorrhiza Bge. and Ligusticum chuanxiong Hort., both of which exert neuroprotective effects. Nevertheless, the effect of GXN on AD is unknown. In the present study, we investigated whether GXN alters cholesterol, amyloid-beta (Aß), gut microbiota, serum metabolites, oxidative stress, neuronal metabolism activities, and apoptosis in an AD model rabbit fed a 2% cholesterol diet. Our results suggested that the GXN treatment significantly reduced cholesterol levels and Aß deposition and improved memory and behaviors in AD rabbits. The 16S rRNA analysis showed that GXN ameliorated the changes in the gut microbiota, decreased the Firmicutes/Bacteroidetes ratio, and improved the abundances of Akkermansia and dgA-11_gut_group. 1H-NMR metabolomics found that GXN regulated 12 different serum metabolites, such as low-density lipoprotein (LDL), trimethylamine N-oxide (TMAO), and glutamate (Glu). In addition, the 1H-MRS examination showed that GXN remarkably increased N-acetyl aspartate (NAA) and Glu levels while reducing myo-inositol (mI) and choline (Cho) levels in AD rabbits, consequently enhancing neuronal metabolism activities. Furthermore, GXN significantly inhibited oxidative stress and neuronal apoptosis. Taken together, these results indicate that GXN attenuates AD via improving gut microbiota, host metabolites, and neuronal apoptosis.

Microbial composition and dynamic succession during the Daqu production process of Northern Jiang-flavored liquor in China.

The microbial community structure and succession regularity of six key periods during high-temperature Daqu production were revealed using high-throughput sequencing to explore the factors affecting the flavor formation of Northern Jiang-flavored Baijiu technology. The results showed that among the six Daqu samples, the bacteria mainly included Firmicutes, Actinobacteriota, and Proteobacteria, of which Proteobacteria was the most dominant. The primary fungus was Ascomycota. At the genus level, the primary bacterial groups were Lactobacillus, Weissella, Bacillus, Delftia, Achromobacter, Saccharopolyspora, Thermoactinomyces, Scopulibacillus, Pseudomonas, and Stenotrophomonas. The main fungal groups in the Daqu were Wickerhamomyces, Saccharomycopsis, Thermoascus, and Thermomyces. During the initial stage of Daqu production, the dominant bacteria were Lactobacillus (20.07%) and Weissella (48.30%). As the fermentation temperature of the Daqu increased, Achromobacter, Stenotrophomonas, and Delftia became the dominant bacteria during the first Daqu flipping period, the second Daqu flipping period, and the dry-fire period. During these three periods, many bacteria were eliminated, decreasing the bacterial diversity, while a decline in temperature was evident during the Daqu exit period. After adapting to the high-temperature environment, the accumulation of Saccharopolyspora (22.07%), Thermoactinomyces (16.73%), Scopulibacillus (27.13%), Kroppenstedtia (9.03%), and Bacillus (6.97%) increased the bacterial diversity during the Daqu exit period. Wickerhamomyces (83.47%) represented the main dominant fungus during the initial production stage but were eliminated with increased temperature. Furthermore, a higher temperature increased the abundance of Saccharomycopsis and Thermoascus, while Thermomyces gradually accumulated in the D, E, and F samples. Thermomyces (79.90%) and Thermoascus (13.83%) became the dominant fungi during the Daqu exit period. In this study, high-throughput sequencing technology was used to reveal the microbial diversity during the high-temperature Daqu production process of Northern Jiang-flavored Baijiu. This provided a scientific basis for improving the production process of this product in the future. Therefore, understanding the formation of the flavor substances and the related microorganisms in Northern Jiang-flavored Baijiu can provide guidance for using them to manipulate the preparation process while implementing microbial control and improving the production procedures. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13205-021-02779-8.

Anodic concentration loss and impedance characteristics in rotating disk electrode microbial fuel cells.

A rotating disk electrode (RDE) was used to investigate the concentration loss and impedance characteristics of anodic biofilms in microbial fuel cells (MFCs). Amperometric time-current analysis revealed that at the rotation rate of 480 rpm, a maximum current density of 168 µA cm(-2) can be achieved, which was 22.2 % higher than when there was no rotation. Linear sweep voltammetry and electrochemical impedance spectroscopy tests showed that when the anodic potential was set to -300 mV vs. Ag/AgCl reference, the power densities could increase by 59.0  %, reaching 1385 mW m(-2), the anodic resistance could reduce by 19  %, and the anodic capacitance could increase by 36 %. These results concur with a more than 85 % decrease of the diffusion layer thickness. Data indicated that concentration loss, diffusion layer thickness, and the mixing velocity play important roles in anodic resistance reduction and power output of MFCs. These findings could be helpful to the design of future industrial-scale MFCs with mixed bacteria biofilms.

Association Between Thyroid Hormones, Lipids and Oxidative Stress Markers in Subclinical Hypothyroidism.

BACKGROUND: Oxidative stress plays a role in the pathogenesis of many chronic diseases. It is recognized in overt hypothyroidism while its existence in subclinical hypothyroidism (SCH) is not well established. The aim of this study was to determine whether there was increased oxidation of lipids and proteins in SCH, and examine their association with lipids and thyroid hormones. METHODS: Male adults (35-59 years) with SCH (n=467) and euthyroid controls (n=190) were studied. Anthropometric measurements, plasma lipids, thyroid stimulating hormone (TSH), free thyroxine (FT4), free triiodothyronine (FT3), total antioxidant capacity (T-AOC), lipid peroxidation products, malondialdehyde (MDA), advanced oxidation protein products (AOPP) and dityrosine concentrations were measured. RESULTS: Plasma concentrations of MDA were significantly higher (p<0.05) in SCH (8.11±1.39 nmol/mL) compared with euthyroid controls (7.34±1.31 nmol/mL) while AOPP, dityrosine and T-AOC levels were not different. MDA was not associated with TSH (ß=-0.019, P=0.759), FT4 (ß=-0.062, P=0.323) and FT3 (ß=-0.018, P=0.780) in SCH while levels increased with elevated total cholesterol (ß=0.229, P=0.001), LDL (ß=0.203, P=0.009) and triglycerides (ß=0.159, P=0.036) after adjustment for age and body mass index. T-AOC reduced (ß=-0.327, P=0.030) with increased MDA in euthyroid controls and not in SCH (ß=-0.068, P=0.349), while levels increased with elevated triglycerides in both groups. CONCLUSIONS: Oxidative stress was increased in subclinical hypothyroidism as evidenced by the elevated lipid peroxidation product, malondialdehyde, while protein oxidation was absent. Thus, reduction of oxidative stress may be beneficial in patients with subclinical hypothyroidism.

Elevated plasma dityrosine in patients with hyperlipidemia compared to healthy individuals.

BACKGROUND: Dityrosine, the modification of tyrosine residues, may contribute to metabolic disorders. This study was undertaken to investigate plasma dityrosine concentrations in patients with hyperlipidemia and to examine the correlation between dityrosine and lipid profiles. METHODS: Fluorescence spectrophotometry was used to measure dityrosine in the plasma of healthy subjects (n = 203) and dyslipidemic subjects, which included patients with mild hyperlipidemia (n = 246) and hyperlipidemia (n = 179). Advanced oxidation protein products (AOPP) and malondialdehyde (MDA) were also assayed in all subjects. RESULTS: Dityrosine levels were higher by 9.3 and 22.9% in mildly hyperlipidemic and hyperlipidemic patients, respectively, compared to controls after adjustment for age, gender, and BMI. AOPP and MDA levels showed similar trends. The levels of dityrosine related positively (p < 0.05) to total cholesterol (r = 0.362), triglycerides (r = 0.449), and low-density lipoprotein cholesterol (r = 0.359). Moreover, plasma dityrosine (r = 0.408), AOPP (r = 0.488), and MDA (r = 0.181) levels were elevated with an increase in the atherosclerosis index in the subjects. CONCLUSIONS: These findings suggest that dityrosine formation may be an early event in the pathological process of hyperlipidemia. Dityrosine as a biomarker detected by fluorescence spectrophotometry might be a useful tool to evaluate the plasma redox state in hyperlipidemia.

Disparities in the Prevalence of Metabolic Syndrome (MS) and its Components Among University Employees by Age, Gender and Occupation.

BACKGROUND: Metabolic Syndrome (MS), a known risk factor for Cardiovascular Disease (CVD) and type II diabetes is an emerging epidemic in China. Studies carried out on the general population indicate a varied clustering of cardiovascular risks in many parts of the country. However, there is limited data on its prevalence in the working population. Workplace can serve as an important place for prevention, control and management of CVD risks. The aim of this study was to investigate the prevalence of MS and its components among University workers, and determine how the prevalence varied according to sex and occupation. METHODS: This was a cross-sectional study of 2,428 University employees (22-60 years) who received an annual clinical examination at the University hospital. Anthropometric measurements, blood pressure, Fasting Plasma Glucose (FPG), and lipid profiles were measured. MS was defined according to the National Cholesterol Education Program Adult Treatment panel III modified criteria. RESULTS: Overall prevalence of MS was 6.1%, higher in males (5.1%) than females (1.1%), and increased with age. The most prevalent MS components in all workers were hypertension (37.9%) and hypertryglyceridemia (20.8%), corresponding rates in males were 28.3% and 16.1% while females had a prevalence of 9.6% and 4.7%. After adjustment for age, administrative work was associated (p<0.05) with increased hypertension (odds ratio (OR) =1.474; 95% confidence interval (CI), 1.146-1.896) and hyperglycemia (OR=1.469; 95% CI, 1.082-1.993) in male workers, and with hypertension (OR=1.492; 95% CI, 1.071-2.080) in females. However, prevalence of hypertryglyceridemia was lower (OR=0.390; 95% CI, 0.204-0.746) in female administrators compared to those in academics. Obesity, MS and reduced High Density Lipoprotein (HDL) cholesterol prevalence was not different (p>0.05) between the two occupations in both sexes. CONCLUSIONS: Prevalence of MS and its components was higher in male workers than in females, increased with age, and were more common in administrative workers. The findings support the need for gender and occupation specific health interventions to prevent CVDs and type II diabetes in the workplace.

Evaluation of the relationship between subclinical hypothyroidism and metabolic syndrome components among workers.

OBJECTIVES: Both hyperthyroidism and overt hypothyroidism are associated with increased prevalence of metabolic syndrome and its components, while data on subclinical hypothyroidism is currently limited especially in working populations. The aim of this study was to examine the association between subclinical hypothyroidism and metabolic syndrome components in workers; and to evaluate whether there are differences by sex and occupation. MATERIAL AND METHODS: A total of 1150 university employees (male - 792, female - 358) aged 30-60 years who came for an annual medical check-up were studied. Anthropometric measurements were taken, and blood pressure, fasting plasma glucose (FPG), lipid profiles, thyroid stimulating hormone (TSH), free thyroxin (FT4) and free triiodothyronine (FT3) levels were measured. RESULTS: After adjustment for age and body mass index (BMI), TSH was positively associated with increased triglyceride (TG) levels (ß = 0.108, p = 0.020) and FPG (ß = 0.130, p = 0.006) in subclinical hypothyroid male workers. However, TSH was not associated (p > 0.05) with any component of metabolic syndrome (MS) in the euthyroid group. In females, TSH was not correlated with MS components in both euthyroid and subclinical hypothyroid groups. Furthermore, comparison by occupation showed higher TSH in subclinical hypothyroid male workers employed in administration (5.23 ± 0.52 mU/l) than those working as academics (5.12 ± 0.52 mU/l), which resulted in elevated systolic and diastolic blood pressure, FPG, total cholesterol, TG and high density lipoprotein cholesterol. In females, BMI, systolic and diastolic blood pressure, TG and FPG were significantly (p < 0.05) higher in subclinical hypothyroid administrators than those in academics. CONCLUSIONS: Subclinical hypothyroidism was associated with metabolic syndrome components in male workers and not in females. Administration workers showed increased metabolic risks compared to academics. The findings suggest that the assessment of thyroid function in individuals with metabolic syndrome in the workplace may be favorable especially among men.