[Effect of Gegen Qinlian Decoction on methylation and expression of Scd1gene in adipose tissue of insulin resistant rats and correlations between methylation and physiological and biochemical parameters].

This study aimed to explore the effect of Gegen Qinlian Decoction(GQD) on the methylation and mRNA expression level of stearoyl CoA desaturase(SCD) gene in the adipose tissue of rats with insulin resistance(IR) induced by high-fat diet as well as the correlations between methylation and physiological and biochemical indicators. The animals were divided into seven groups, namely, blank control(C) group, IR model group, low-(1.65 g·kg~(-1)), medium-(4.95 g·kg~(-1)), and high(14.85 g·kg~(-1))-dose GQD(GQDL, GQDM, and GQDH) groups, rosiglitazone(RGN, 5 mg·kg~(-1)) group, and simvastatin(SVT, 10 mg·kg~(-1)) group. The rat epididymal adipose tissue was collected for detecting all the cytosine methylation levels in two fragments of Scd1 gene by bisulfite sequencing PCR(BSP). Scd1-1 was located in CG shores and Scd1-2 in CG islands, including the transcriptional start site(TSS). The Scd1 mRNA level was determined by quantitative real-time PCR(q-PCR). Spearman correlation coefficient was used to analyze the correlations between amplified fragment C methylation and physiological and biochemical indicators. The results showed that GQDM remarkably reversed the elevated CG7 methylation in the TSS upstream region of Scd1-2 triggered by high-fat diet. GQDL significantly reversed the lowered total CG methylation in the downstream region of Scd1-2 induced by the high-fat diet. GQD did not significantly improve the decreased Scd1 mRNA expression caused by high-fat diet. Changes in methylation of the total CG, CG5 and CT11 of Scd1-1 in CG shores exhibited significant negative correlations with the serum triglyceride(TG) but positive correlation with the Scd1 mRNA level. The methylation of several C sites in the TSS upstream region of Scd1-2 was positively correlated with physiological and biochemical parameters. The methylation of several CG sites in the TSS downstream region of Scd1-2 was negatively associated with physiological and biochemical parameters. Besides, the methylation of several CH sites in the downstream fragment was positively correlated with physiological and biochemical parameters. All these have demonstrated that GQD may exert the therapeutic effect by regulating the methylation of CG7 in the TSS upstream region and total CG site in the TSS downstream region of Scd1 gene. The methylation of total CG, CG5 and CT11 sites in CG shores of Scd1 gene may be important targets for regulating Scd1 mRNA level and affecting serum TG.

[Research on XOD captured components in Lagotis brevituba based on UPLC-Q-TOF-MS and molecular docking technology].

Potential xanthine oxidase (XOD) inhibitors in Lagotis brevituba were captured by using affinity and ultrafiltration. The structures of the captured components were identified by ultra-performance liquid chromatography coupled with Q-TOF mass spectrometry (UPLC-Q-TOF-MS). The binding intensity and binding mechanism between the captured components and XOD were analyzed by using molecular docking software Autodock 4.2. A total of 17 compounds were identified, including 9 flavonoids, 5 phenolic acids and 3 triterpenes. Molecular docking results showed that all the captured components could be spontaneously bound with XOD mainly via hydrogen bond, Van der Waals' force and hydrophobic interaction. From the perspective of binding energy and scoring function, the collected fractions all had potential prospects for XOD inhibitors, and the flavonoid luteolin-3',7 glucuronide had the best effect. The results also showed that affinity and ultrafiltration, ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) and molecular docking technology can provide a powerful tool for the analysis of XOD inhibitor components in natural products.

Incidence, impact and predictors of residual device patency after left atrial appendage closure with the LACbes device.

BACKGROUND: The residual device patency (RDP) after left atrial appendage closure (LAAC) with the LACbes device has not been specifically explored in atrial fibrillation (AF) patients. This study aims to explore the incidence, impact and predictors of RDP detected by cardiac computed tomography angiography (CCTA) post LAAC. METHODS: AF patients implanted with the LACbes device were prospectively enrolled. CCTA device surveillance was performed at 3 months post-procedure. Major adverse events (MAEs), including stroke/transient ischemic attack, major bleeding and all-cause death, were evaluated. RESULTS: Among 141 patients with CCTA surveillance, 56 (39.7%) showed no visible leak and 85 (60.3%) showed RDP. During the median follow-up of 443 [232, 706] days, the presence of RDP was not associated with an increased risk of MAEs (adjusted hazard ratio [HR]: 4.07, 95% confidence interval [CI]: 0.49-34.24, p = 0.196), while peri-device leak (PDL) at the lobe was associated with heightened risks of MAEs (adjusted HR: 6.85, 95% CI: 1.62-28.89, p = 0.009). In patients with PDL at the lobe, antiplatelet after 6 months (HR: 0.20, 95% CI: 0.05-0.91, p = 0.038) was independent protective predictor of MAEs. Besides, current smoking (odds ratio [OR]: 7.52, 95% CI: 2.68-21.08, p < 0.001) and maximum diameter of LAA orifice (OR: 1.16, 95% CI: 1.00-1.34, p = 0.048) were independent predictors of PDL at the lobe. CONCLUSIONS: Presence of PDL at the device lobe detected by CCTA at 3-month post LAAC with LACbes is associated with unfavorable prognosis in AF patients. CLINICAL TRIAL REGISTRATION: ClinicalTrials.gov Identifier: NCT03788941.

Prognostic Impact of Left Atrial Appendage Patency After Device Closure.

BACKGROUND: The prognostic impact of left atrial appendage (LAA) patency, including those with and without visible peri-device leak (PDL), post-LAA closure in patients with atrial fibrillation, remains elusive. METHODS: Patients with atrial fibrillation implanted with the WATCHMAN 2.5 device were prospectively enrolled. The device surveillance by cardiac computed tomography angiography was performed at 3 months post-procedure. Adverse events, including stroke/transient ischemic attack (TIA), major bleeding, cardiovascular death, all-cause death, and the combined major adverse events (MAEs), were compared between patients with complete closure and LAA patency. RESULTS: Among 519 patients with cardiac computed tomography angiography surveillance at 3 months post-LAA closure, 271 (52.2%) showed complete closure, and LAA patency was detected in 248 (47.8%) patients, including 196 (37.8%) with visible PDL and 52 (10.0%) without visible PDL. During a median of 1193 (787-1543) days follow-up, the presence of LAA patency was associated with increased risks of stroke/TIA (adjusted hazard ratio for baseline differences, 3.22 [95% CI, 1.17-8.83]; P=0.023) and MAEs (adjusted hazard ratio, 1.12 [95% CI, 1.06-1.17]; P=0.003). Specifically, LAA patency with visible PDL was associated with increased risks of stroke/TIA (hazard ratio, 3.66 [95% CI, 1.29-10.42]; P=0.015) and MAEs (hazard ratio, 3.71 [95% CI, 1.71-8.07]; P=0.001), although LAA patency without visible PDL showed higher risks of MAEs (hazard ratio, 3.59 [95% CI, 1.28-10.09]; P=0.015). Incidences of stroke/TIA (2.8% versus 3.0% versus 6.7% versus 22.2%; P=0.010), cardiovascular death (0.9% versus 0% versus 1.7% versus 11.1%; P=0.005), and MAEs (4.6% versus 9.0% versus 11.7% versus 22.2%; P=0.017) increased with larger PDL (0, >0 to ≤3, >3 to ≤5, or >5 mm). Older age and discontinuing antiplatelet therapy at 6 months were independent predictors of stroke/TIA and MAEs in patients with LAA patency. CONCLUSIONS: LAA patency detected by cardiac computed tomography angiography at 3 months post-LAA closure is associated with unfavorable prognosis in patients with atrial fibrillation implanted with WATCHMAN 2.5 device. REGISTRATION: URL: https://www.clinicaltrials.gov; Unique identifier: NCT03788941.

Efficacy of Left Atrial Appendage Closure and Oral Anticoagulation After Atrial Fibrillation Catheter Ablation.

Left atrial appendage closure (LAAC) proved to be noninferior to oral anticoagulation (OAC) in nonablated patients with atrial fibrillation (AF). This study aimed to compare the efficacy and safety of LAAC with those of OAC therapy in patients after AF ablation. This study included patients who underwent catheter ablation (CA) of AF between January 2016 and December 2020. The cohort was divided into CA + LAAC and CA + OAC, where propensity score matching was used to select controls, and each group contained 682 subjects. The enrolled patients' mean age was 70.34 ± 8.32 years, and 47.3% were female; their CHA2DS2-VASc score was 3.48 ± 1.17. Baseline characteristics were similar between groups. After a 3-year mean follow-up, the incidence of thromboembolic events was 1.25 and 1.10 and that of major bleeding events was 0.65 and 1.72 per 100 patient-years in the CA + LAAC, and CA + OAC groups, respectively. The rate of thromboembolisms and major adverse cardiovascular events was similar between the 2 groups (hazard ratio [HR] 1.162, 95% confidence interval [CI] 0.665 to 2.030, p = 0.598, HR 0.711, 95% CI 0.502 to 1.005, p = 0.053); however, that of major bleeding and all-cause death was significantly reduced with LAAC (HR 0.401, 95% CI 0.216 to 0.746, p = 0.004, HR 0.528, 95% CI 0.281 to 0.989, p = 0.046). There was no significant difference in periprocedural complications (p >0.05) and the rate of AF recurrence (OAC vs LAAC: 39.44% vs 40.62%, p = 0.658). LAAC is a reasonable and safer alternative to OAC therapy in high-risk patients after AF ablation.

[Effects of Wastewater Discharge on Antibiotic Resistance Genes and Microbial Community in a Coastal Area].

Wastewater treatment plants (WWTPs) are important sources of antibiotic resistance genes (ARGs) in aquatic environments. Mobile genetic elements (MGEs) and microbial communities are key factors that affect the proliferation of ARGs. To reveal the effects of WWTPs effluent discharge on the ARGs and microbial community in a coastal area, the structure and distribution of ARGs, MGEs, and microbial community in Shangyu (SY) and Jiaxing (JX) effluent receiving areas (ERAs) and the offshore area of Hangzhou Bay (HB) were investigated via high-throughput quantitative PCR and 16S rRNA high-throughput sequencing. The results showed that multidrug resistance genes were the most abundant ARGs across all the sampling sites. The diversity and abundance of ARGs and MGEs in the ERAs were much higher than those in the HB. Additionally, the diversities of the microbial community in the JX-ERA were higher than those in the SY-ERA and HB. PCoA showed that the distribution of ARGs, MGEs, and microbial communities in the ERAs and HB were significantly different, indicating that the long-term wastewater discharge could alter the distribution of ARGs, MGEs, and microbial communities in the coastal area. The co-occurrence pattern among ARGs, MGEs, and microbial communities revealed that 12 bacterial genera, such as Psychrobacter, Pseudomonas, Sulfitobacter, Pseudoalteromonas, and Bacillus, showed strong positive correlations with ARGs and MGEs. Most potential hosts carried multidrug and ß-lactamase resistance genes.

[Distribution Characteristics of Antibiotics and Antibiotic Resistance Genes in Wastewater Treatment Plants].

Antibiotics and antibiotic resistance genes (ARGs) are highly concerning emerging polltants. Wastewater treatment plants (WWTPs) are considered to be one of the most important anthropogenic sources of antibiotics and ARGs in the environment. Six WWTPs of a city were selected to investigate the antibiotics and ARGs in the influents and effluents, respectively. In total, 73 kinds antibiotics belonging to 7 classes were detected in WWTPs. The total concentration of those antibiotics ranged between 490.2-2288.6 ng·L-1, and ofloxacin, maprofloxacin, and roxithromycin were the most frequently detected antibiotics in the influents. A dramtic difference was observed in the antibiotic removal efficiency among the WWTPs, and the total antibiotic concentration in the effluents ranged between 260.2-1464.3 ng·L-1. Macrolides and quinolones are the main classes in the effluents, and the antibiotic concentration in the effluents was higher than that in the influents. Ten kinds of ARGs were detected in the WWTPs. Among these, sul Ⅰ was the most widely prevalent with an absolute abundance of 2.4×105-5.4×106 copies·mL-1. No significant difference in ARG abundance was observed between industrial wastewater and domesic wastewater. Furthermore, the correlation between ARG abundance and antibiotic concentrations weakened after treatment in the WWTPs.

[Distribution Characteristics of Antibiotic Resistance Genes in Wastewater Treatment Plants].

Wastewater treatment plants (WWTPs) are regarded as the main source for antibiotic resistant genes (ARGs). To explore the features regarding the distribution of ARGs in wastewater with complicated composition in treatment plants, wastewater samples from a chemical industry park that produced antibiotics were selected. qPCR was applied to detect the type and abundance of ARGs in the wastewater flows from the WWTPs. The results indicated that 16 types of ARGs were detected from the wastewater from the WWTPs, among which sulfonamide resistance genes and tetracycline resistance genes were the dominant ARGs that appeared in the wastewater. Additionally, intI 1 was detected and its abundance was correlated with that of sulfonamide resistance genes. This indicated that intI 1 may promote the migration and transformation of sulfonamide resistance genes. The pharmaceutical factories in the park mainly synthesize macrolide antibiotics. Because of the selective pressure, the absolute abundance of ermB in the wastewater was much higher than that in the other industrial wastewater. The total ARGs decreased by 1.16 log via traditional biological treatment process, and the total ARGs decreased by 2.46 log via the Fenton process. The results showed that the removal effects of deep treatment processes on ARGs were better than that of biological treatment in this wastewater treatment process. Highly abundant and movable ARGs already exist in the water body, and their release from WWTPs without effective treatment poses high risks to the environment.

Evaluating sampling strategy for DNA barcoding study of coastal and inland halo-tolerant Poaceae and Chenopodiaceae: A case study for increased sample size.

Environmental conditions in coastal salt marsh habitats have led to the development of specialist genetic adaptations. We evaluated six DNA barcode loci of the 53 species of Poaceae and 15 species of Chenopodiaceae from China's coastal salt marsh area and inland area. Our results indicate that the optimum DNA barcode was ITS for coastal salt-tolerant Poaceae and matK for the Chenopodiaceae. Sampling strategies for ten common species of Poaceae and Chenopodiaceae were analyzed according to optimum barcode. We found that by increasing the number of samples collected from the coastal salt marsh area on the basis of inland samples, the number of haplotypes of Arundinella hirta, Digitaria ciliaris, Eleusine indica, Imperata cylindrica, Setaria viridis, and Chenopodium glaucum increased, with a principal coordinate plot clearly showing increased distribution points. The results of a Mann-Whitney test showed that for Digitaria ciliaris, Eleusine indica, Imperata cylindrica, and Setaria viridis, the distribution of intraspecific genetic distances was significantly different when samples from the coastal salt marsh area were included (P < 0.01). These results suggest that increasing the sample size in specialist habitats can improve measurements of intraspecific genetic diversity, and will have a positive effect on the application of the DNA barcodes in widely distributed species. The results of random sampling showed that when sample size reached 11 for Chloris virgata, Chenopodium glaucum, and Dysphania ambrosioides, 13 for Setaria viridis, and 15 for Eleusine indica, Imperata cylindrica and Chenopodium album, average intraspecific distance tended to reach stability. These results indicate that the sample size for DNA barcode of globally distributed species should be increased to 11-15.

[Conversion Pathways of Substrates in Sulfate-Reducing Ammonia Oxidation System].

The phenomenon of simultaneous transformation of ammonium and sulfate under the conditions of inoculating ANAMMOX culture has gotten the attention of researchers. However, there are some problems and doubts reported in the related literature. In this study, the characteristics of ammonium and sulfate synchronous transformation were investigated in a CFSTR via inoculation with ANAMMOX culture. Under the condition of oxygen removal and non-filling, in the unfilled sealed fermentation tank, the average conversion of NH4+-N was 50.8 mg·L-1 while that of sulfate-sulfur was 4.5 mg·L-1. Elemental analysis results showed that the observed yellow solid was not elemental sulfur but rather iron-containing compounds. However, no obvious change of ammonium was observed when using a filled sealed batch reactor. Only sulfate transformed significantly, and the transformation rate was affected by the inoculation biomass. Under these two conditions, the ORP in the reactor was completely different. This is an indication that the synchronous transformation of ammonium and sulfate observed in both our study and other related studies is probably not a process mediated by ANAMMOX organisms, in which sulfate acted as the electron acceptor to oxidize ammonium. Actually, ammonium and sulfate transformation were completely independent: ammonium oxidation is due to the micro oxygen environment created by the reactor operation form, whereas sulfate conversion is attributed to the sulfate heterotrophic reduction that results from the organic matter release via microbial decay. This transformation can clarify and explain the problems and doubts reported in the related research.

[Biotransformation of Nitrate to Nitrogen Gas Driven by ANAMMOX Microbes via Zero-valent Iron Under Anaerobic Conditions].

A novel type of nitrate removal process was investigated in this study, which coupled nitrate reduction by zero-valent iron(ZVI) with anaerobic ammonia oxidation(AMAMMOX). The zero-valent iron dosage was 71 g·L-1. The inoculation volume of ANAMMOX granular sludge was 200 mg·L-1. The agitation rate and environment reaction temperature of the CSTR was set up to 150 r·min-1 and 33℃±0.5℃, respectively. The pH of influent was maintained at 7.0-8.0.The result indicated that ANAMMOX bacteria could survive in nitrate and ZVI supplied system under neutral condition. The reactor was set up quickly with the nitrogen removal rate of 0.12 kg·(m3·d)-1 on the first 5 days, and maintained over 0.1 kg·(m3·d)-1 nitrogen removal ability with the effluent nitrite and ammonia concentrations below 2.0 mg·L-1 and pH value higher than 8.0. The nitrogen loss was up to 89% under the condition of 30-40℃, pH 4-6.Too extreme pH and temperature conditions were not favorable for the coupling reactions.