Liraglutide attenuates type 2 diabetes mellitus-associated non-alcoholic fatty liver disease by activating AMPK/ACC signaling and inhibiting ferroptosis.

BACKGROUND: Non-alcoholic fatty liver disease (NAFLD) is one of the most common complications of type 2 diabetes mellitus (T2DM). The pathogenesis of NAFLD involves multiple biological changes, including insulin resistance, oxidative stress, inflammation, as well as genetic and environmental factors. Liraglutide has been used to control blood sugar. But the impact of liraglutide on T2DM-associated NAFLD remains unclear. In this study, we investigated the impact and potential molecular mechanisms of inhibiting ferroptosis for liraglutide improves T2DM-associated NAFLD. METHODS: Mice were fed on high-fat-diet and injected with streptozotocin to mimic T2DM-associated NAFLD and gene expression in liver was analysed by RNA-seq. The fast blood glucose was measured during the period of liraglutide and ferrostatin-1 administration. Hematoxylin and eosin staining was used to evaluate the pathological changes in the liver. The occurrence of hepatic ferroptosis was measured by lipid peroxidation in vivo. The mechanism of liraglutide inhibition ferroptosis was investigated by in vitro cell culture. RESULTS: Liraglutide not only improved glucose metabolism, but also ameliorated tissue damage in the livers. Transcriptomic analysis indicated that liraglutide regulates lipid metabolism related signaling including AMPK and ACC. Furthermore, ferroptosis inhibitor rather than other cell death inhibitors rescued liver cell viability in the presence of high glucose. Mechanistically, liraglutide-induced activation of AMPK phosphorylated ACC, while AMPK inhibitor compound C blocked the liraglutide-mediated suppression of ferroptosis. Moreover, ferroptosis inhibitor restored liver function in T2DM mice in vivo. CONCLUSIONS: These findings indicate that liraglutide ameliorates the T2DM-associated NAFLD, which possibly through the activation of AMPK/ACC pathway and inhibition of ferroptosis.

A conversion CRT strategy combined with AVJA may be a perspective alternative for heart failure patients with persistent atrial fibrillation.

Heart failure (HF) combined with persistent atrial fibrillation (AF) often coexist and may promote the pathological conditions of cardiac dysfunction, leading to poor prognosis. Cardiac resynchronization therapy (CRT) combined with atrioventricular junction ablation (AVJA) is a highly effective treatment for HF patients with underlying AF who either have failed or are not suitable for catheter ablation. The CRT-AVJA combination therapy can improve clinical outcomes in HF patients. Currently, clinical CRT methods are categorized into biventricular pacing (BVP) - based and conduction system pacing (CSP) - based methods. These procedures have inherent advantages and disadvantages, in addition to their considerable differences in clinical applications. This article aims to review the clinical progress of AVJA combined with different CRT strategies for treating HF patients with persistent AF and propose that conversion CRT strategy (BVP/CSP-CRT) combined with AVJA may be a perspective alternative. Meanwhile, we generalize that 7 categories of HF patients with persistent AF may need to consider the CRT-AVJA combination therapy.

Identification and evaluation of reference genes for quantitative real-time PCR analysis in Passiflora edulis under stem rot condition.

Passion fruit (Passiflora edulis), an important tropical and subtropical fruit, has a high edible and medicinal value. Stem rot disease is one of the most important diseases of passion fruit. An effective way for control and prevention of this disease is to identify the genes associated with resistance to this disease. Quantitative real-time PCR (RT-qPCR) has mainly been widely applied to detect gene expression because of its simplicity, fastness, low cost and high sensitivity. One of the requirements for RT-qPCR is the availability of suitable reference genes for normalization of gene expression. However, currently, no Passiflora edulis reference genes have been identified andthus it has hindered the gene expression studies in this plant. The present study aimed to address this issue. We analyzed sixteen candidate reference genes, including nine common (GAPDH, UBQ, ACT1, ACT2, EF-1α-1, EF-1α-2, TUA, NADP, and GBP) and seven novel genes (C13615, C24590, C27182, C10445, C21209, C22199, and C22526), in different tissues (stem, leaf, flower and fruit) of two accessions under stem rot condition. We calculated the expression stability in twenty-four samples using the ΔCt, GeNorm, NormFinder, BestKeeper and RefFinder. The results showed that both C21209 and EF-1α-2 were sufficient to normalize gene expression under stem rot, whereas the commonly used reference genes, GAPDH and UBQ, were the least stable ones. The expression patterns of PeUFC under stem rot condition normalized by stable and unstable reference genes indicated the suitability of using the optimal reference genes. To our knowledge, this is the first systematic study of reference genes in Passiflora edulis, which identified a number of reliable reference genes suitable for gene expression studies in Passiflora edulis by RT-qPCR.

Effect of biodegradable polymer drug-eluting stents versus biocompatible polymer everolimus-eluting stents: a meta-analysis.

OBJECTIVE: Biocompatible polymer everolimus-eluting stents (EES) are associated with risk of stent thrombosis (ST); biodegradable polymer drug-eluting stents (BP-DES) were designed to reduce these risks. However, the long-term benefits are not completely clear. METHOD: We undertook a meta-analysis of randomized studies identified in systematic searches of MEDLINE, EMBASE, and the Cochrane Database. Primary outcome was the risk of ST. RESULTS: Twelve studies (11,692 patients) were included. Overall, compared with EES, BP-DES were associated with a broadly equivalent risk of definite and probable ST (OR, 0.91; 95% CI, 0.55 to 1.50; P = 0.71; I2 = 0.0%), early ST (OR, 2.25; 95% CI, 0.78 to 6.47; P = 0.13; I2 = 0.0%), late ST (OR, 3.57; 95% CI, 0.42 to 30.58; P = 0.25; I2 = 0.0%) and very late ST (OR, 0.50; 95% CI, 0.05 to 5.52; P = 0.57). Meanwhile, there was no significant difference in all-cause mortality (OR, 1.07; 95% CI, 0.86 to 1.32; P = 0.54; I2 = 0.0%), myocardial infarction (OR, 1.07; 95% CI, 0.88 to 1.30; P = 0.47; I2 = 0.0%), target vessel revascularization (OR, 1.02; 95% CI, 0.86 to 1.21; P = 0.80; I2 = 12.0%), and major adverse cardiac events (OR, 1.04; 95% CI, 0.93 to 1.16; P = 0.53; I2 = 0.0%). Furthermore, angiographic data showed that in-stent and in-segment late luminal loss were similar between the two groups. CONCLUSIONS: Compared with biocompatible polymer EES, biodegradable polymer stents appear to have equivalent clinical benefits.

Contribution of atmospheric nitrogen deposition to diffuse pollution in a typical hilly red soil catchment in southern China.

Atmospheric nitrogen (N) deposition is currently high and meanwhile diffuse N pollution is also serious in China. The correlation between N deposition and riverine N export and the contribution of N deposition to riverine N export were investigated in a typical hilly red soil catchment in southern China over a two-year period. N deposition was as high as 26.1 to 55.8kgN/(ha·yr) across different land uses in the studied catchment, while the riverine N exports ranged from 7.2 to 9.6kgN/(ha·yr) in the forest sub-catchment and 27.4 to 30.3kgN/(ha·yr) in the agricultural sub-catchment. The correlations between both wet N deposition and riverine N export and precipitation were highly positive, and so were the correlations between NH4(+)-N or NO3(-)-N wet deposition and riverine NH4(+)-N or NO3(-)-N exports except for NH4(+)-N in the agricultural sub-catchment, indicating that N deposition contributed to riverine N export. The monthly export coefficients of atmospheric deposited N from land to river in the forest sub-catchment (with a mean of 14%) presented a significant positive correlation with precipitation, while the monthly contributions of atmospheric deposition to riverine N export (with a mean of 18.7% in the agricultural sub-catchment and a mean of 21.0% in the whole catchment) were significantly and negatively correlated with precipitation. The relatively high contribution of N deposition to diffuse N pollution in the catchment suggests that efforts should be done to control anthropogenic reactive N emissions to the atmosphere in hilly red soil regions in southern China.

Interaction between the hydrochemical environment, dissolved organic matter, and microbial communities in groundwater: A case study of a vegetable cultivation area in Huaibei Plain, China.

Intensive vegetable planting has a profound impact on the surrounding aquatic environment. The self-purification ability of groundwater is poor, and it is difficult to return groundwater to its original state once polluted. Therefore, it is necessary to clarify the impact of intensive vegetable planting on groundwater. This study selected the groundwater of a typical intensive vegetable planting base in the Huaibei Plain of China as the research object. This work analyzed the content of major ions, the dissolved organic matter (DOM) composition, and the bacterial community structure in groundwater. Redundancy analysis was used to explore the interactions between the major ions, the DOM composition, and the microbial community. The results showed that under the influence of intensive vegetable planting, the F- and NO3--N contents in groundwater were significantly increased; the excitation-emission matrix combined with parallel factor analysis identified four fluorescent components (C1 and C2 were humus-like components, while C3 and C4 were protein-like components), which mainly consisted of protein-like components. Proteobacteria was the dominant phylum (mean = 69.27 %), followed by Actinobacteriota (mean = 7.25 %) and Firmicutes (mean = 4.02 %), which together explained over 80 % of the total abundance; and TDS, pH, K+, and C3 were the main influencing factors affecting the microbial community structure. This study provides a better understanding of the impact of intensive vegetable cultivation on groundwater.

Effect of biochar application rate on changes in soil labile organic carbon fractions and the association between bacterial community assembly and carbon metabolism with time.

Biochar aging affects the stability of soil carbon. Analyzing the effect of biochar on soil organic carbon (SOC) forms and their relations with microbial community assembly and carbon metabolism with time is helpful for soil carbon sequestration (by adapting the farm management approach). Four treatments with no, low, medium, and high biochar application rates (0 %, 1 %, 2 %, and 4 % of the total dry weight of topsoil before winter wheat planting, abbreviated as control, LB, MB, and HB, respectively) were conducted in the field. The SOC and particulate organic carbon positively correlated with the biochar application rate. Biochar decreased readily oxidizable carbon (P < 0.05) after 8 months of application compared to the control; however, the difference disappeared with time. Biochar increased dissolved organic carbon (DOC) but had no effect on water- soluble organic carbon (WSOC); DOC and WSOC decreased with time. Furthermore, LB and HB stabilized the bacterial alpha diversities with time. Based on high-throughput sequencing, HB reduced the relative abundance of Actinobacteriota but increased that of Acidobacteria (P < 0.05) after 12 months of biochar application. Time-wise, the bacterial community assembly was determined by deterministic processes that were significantly affected by the available nitrogen, DOC, or WSOC. Compared with the control, biochar decreased bacterial links and improved bacterial metabolism of phenolic acids and polymers with time, as evidenced by Biolog EcoPlates. Structural equation modeling revealed that the contribution of bacterial assembly processes to carbon metabolism changed with time. Microbial carbon metabolism was most positively influenced by differences in the composition of bacterial specialists. These findings reinforced that changes in soil labile organic carbon were time-dependent but not necessarilty affected by the biochar application rate.

Comparative transcriptomic analysis reveals the cold acclimation during chilling stress in sensitive and resistant passion fruit (Passiflora edulis) cultivars.

Chilling stress (CS) is an important limiting factor for the growth and development of passion fruit (Passiflora edulis) in winter in South China. However, little is known about how the passion fruit responds and adapts to CS. In this study, we performed transcriptome sequencing of cold-susceptible cultivar Huangjinguo (HJG) and cold-tolerant cultivar Tainong 1 (TN1) under normal temperature (NT) and CS conditions, and a total of 47,353 unigenes were obtained by seven databases. Using differentially expressed unigenes (DEGs) analysis, 3,248 and 4,340 DEGs were identified at two stages, respectively. The Gene Ontology (GO) enrichment analysis showed that the DEGs were mainly related to phosphorylation, membrane protein, and catalytic activity. In Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway, the unigenes of plant-pathogen interaction, plant hormone signal transduction and fatty acid metabolism were enriched. Then, the 12,471 filtered unigenes were clustered into eight co-expression modules, and two modules were correlated with CS. In this two modules, 32 hub unigenes were obtained. Furthermore, the unigenes related to CS were validated using quantitative real-time PCR (RT-qPCR). This work showed that the expression levels of CS-related unigenes were very different in two passion fruit cultivars. The results provide information for the development of passion fruit with increased chilling tolerance.

Complete chloroplast genome sequence of Passiflora serrulata Jacq. (Passifloraceae).

This study was the first report for the complete chloroplast genome of Passiflora serrulata Jacq. (Passifloraceae). The cp genome was 149,683 bp in length contained two inverted repeats (IRs) of 25,470 bp, which were separated by large single-copy (LSC) and small single-copy (SSC) of 86,252 bp and 13,491 bp, respectively. A total of 110 functional genes were encoded, comprised 76 protein-coding genes, 30 tRNA genes, and four rRNA genes. The GC content was 37.0%. The maximum likelihood phylogenetic tree indicated that P. serrulata was recovered as the member of subg. Passiflora and most closely related to the clade formed by P. serratodigitata and P. ligularis.

[Effects of Drip Irrigation Patterns and Biochar Addition on Soil Mineral Nitrogen and Microbial Regulation of Greenhouse].

Drip irrigation and biochar amendment could affect the nitrogen form and transformation. Creating a deep understanding of the interacting effects of drip irrigation patterns and biochar on soil mineral nitrogen, as well as the key functional genes and microbial community involved in nitrogen transformation is helpful for improving facility agricultural management, increasing water and nitrogen use efficiency, and reducing the nitrate accumulation and groundwater pollution caused by nitrogen leaching. Four treatments [surface drip irrigation (D), insert drip irrigation (ID, insert depth 15 cm), surface drip irrigation +10 t·hm-2 of biochar (DB), and insert drip irrigation +10 t·hm-2 of biochar (IDB)] were conducted in a solar greenhouse, and non-rhizospheric and rhizospheric soils of pepper plants were studied. There was no effect of drip irrigation patterns and biochar on ammonium-nitrogen in the non-rhizospheric and rhizospheric soils. Compared with surface drip irrigation, insert drip irrigation decreased the nitrate-nitrogen concentration in the non-rhizosphere soil (P<0.05), but biochar addition weakened the difference. Biochar addition decreased the nitrate-nitrogen concentration in the rhizosphere soil under the same drip irrigation patterns. In the D treatment, biochar significantly decreased the number of copies of AOA, AOB, and nirK genes in the non-rhizospheric soil, and AOA gene copies in the rhizospheric soil (P<0.05); however, there was an increase in the number of copies of AOB and nirK genes in the rhizospheric soil of the D and ID treatments (P<0.05). Based on the structural equation model (SEM), in the non-rhizospheric and rhizospheric soils, pH and electrical conductivity were the environmental factors with the greatest influence on the ammonium-nitrogen and nitrate concentrations, respectively, and the gene copy number of AOB was the biotic factor with the greatest influence on the nitrate-nitrogen concentration. Based on PICRUSt, the γ-Proteobacteria contributed mostly to ammonia monooxygenase gene (K10945) expression, whereas the α-Proteobacteria, especially the rhizobia members, contributed mostly to nitrite reductase gene (K00368) expression. Biochar addition regulated the bacterial community structure that participated in K10945 gene expression in the non-rhizospheric soil and K00368 gene expression in the rhizospheric soil (P<0.05). Overall, biochar addition contributed more to nitrate-nitrogen and microbial mineral nitrogen-transformation processes in the agricultural soil than did the drip irrigation patterns.

[Influence of Biochar Amendment on Soil Denitrifying Microorganisms in Double Rice Cropping System].

At present, it is not explicit how biochar regulates the microbial process of denitrification in paddy fields. Therefore, a field experiment was carried out in a double rice cropping system with three wheat straw biochar treatments:no biochar treatment (CK), added 24 t·hm-2 biochar (LC), and added 48 t·hm-2 biochar (HC). Real time PCR (qPCR) and terminal-restricted fragment length polymorphism (T-RFLP) technology were adopted to analyze the abundances and microbial community structures of denitrification functional genes (narG, nirK and nosZ) in the fallow season and rice season. Due to its alkalinity, biochar amendment increased soil pH by 0.2-0.8. Biochar amendment also increased soil NH4+-N and NO3--N contents by 21.1%-32.5% and 63.0%-176.0% in the fallow season due to the presence of soluble N. Nevertheless, it reduced NH4+-N content by 48.8%-60.1% in the rice season due to the adsorption of biochar. The amendment increased soil MBN content in the fallow season, which may be a result of the large surface of biochar supplying nutrients and a suitable survival environment for the microorganisms. In the fallow season, compared to CK treatment, the increased soil NH4+-N and NO3--N with biochar amendment promoted the conversion of NH4+-N to NO3--N, and thus increased the abundances of narG and nosZ (P<0.05). The higher soil pH with biochar addition increased the abundances of nosZ and altered the community structures of narG and nosZ in the fallow season. Biochar amendment altered the denitrification process, but it did not change N2O emissions in the fallow season, which might reduce NO3--N leaching losses. In the rice season, biochar amendment increased nosZ abundance (P<0.05). HC increased the nirK gene abundance, which contributed to increased N2O emission in the rice season (P<0.05). Biochar converted the community structures of nirK and nosZ by decreasing the NH4+-N content in the rice season. The changes of the narG community structure with HC treatment contributed to the increased N2O emission. In conclusion, biochar amendment can influence the microbes involved in soil denitrification by changing the soil properties, and thus impact the N2O emissions and NO3--N leaching.

[Effects of Biochar Amendment on Soil Microbial Biomass Carbon, Nitrogen and Dissolved Organic Carbon, Nitrogen in Paddy Soils].

Biochar can influence soil microbial biomass. It is not clear how biochar amendment affects soil microbial biomass carbon and nitrogen (MBC and MBN) and dissolved organic carbon and nitrogen (DOC and DON) in double-cropping rice soils. To address this problem, two subtropical double-cropping rice soils (S1 and S2) were selected for an incubation experiment. S1 is developed from granite-weathered red soil and S2 is developed from Quaternary red clay. The following three wheat straw-derived biochar application rates were used, without N fertilizer, in each paddy soil:0%, 1%, and 2% of soil weight, represented by CK, LB, and HB, respectively. After a 70 d incubation, soil mean MBC was 877.03 mg·kg-1, 832.11 mg·kg-1, and 849.30 mg·kg-1 in S1 for the three application rates, and 902.94 mg·kg-1, 874.19 mg·kg-1, and 883.22 mg·kg-1, respectively, in S2. S1+LB, S1+HB, and S2+LB treatments reduced soil mean MBC compared to the CK treatment (P<0.05). This may be attributed to biochar inhibiting microbial growth by adsorbing soil organic carbon and other low-molecular-weight organic matter. Low biochar application rates decreased mean soil MBN by 9.45% compared to the CK treatment in S1 (P<0.05). No significant differences in mean MBC/MBN were observed among the S1 treatments, but LB reduced MBC/MBN in S2 (P<0.05). Due to the soluble organic carbon content and strong alkalinity of biochar, biochar amendment increased mean soil DOC by 4.42%-22.20% and 10.57%-35.47% in S1 and S2, respectively (P<0.05). However, biochar amendment (except for the S2+HB treatment) decreased mean soil DON in both paddy soils. This may have resulted from the adsorption of soil organic nitrogen by biochar and N consumption during the decomposition of the organic carbon within biochar. Biochar amendment increased mean soil DOC/DON in both paddy soils (P<0.05) and mean DOC/DON increased with an increase in the biochar application rate. Based on these results, biochar amendment increased soil dissolved organic carbon, decreased soil microbial biomass, and enhanced the nitrogen deficit in double-cropping paddy soils. Therefore, biochar should be combined with the application with fertilizer in double-cropping rice systems in subtropical central China.

MicroRNA-497 promotes proliferation and inhibits apoptosis of cardiomyocytes through the downregulation of Mfn2 in a mouse model of myocardial ischemia-reperfusion injury.

INTRODUCTION: Myocardial ischemia-reperfusion (I/R) injury affects millions of people worldwide and has a very high mortality rate. Since microRNA-497 (miR-497) has been found to be related with cardiomyocyte apoptosis, this study aimed to explore the effect of miR-497 by targeting Mfn2 in a mouse model of myocardial ischemia-reperfusion (I/R) injury. MATERIALS: BALB/c mice were modeled with I/R and some were injected with miR-497 agomir before I/R to observe whether miR-497 alleviates the injury that occurs as a result of I/R. Bioinformatics website and dual-luciferase reporter gene assay were employed in order to detect the relations between miR-497 and Mfn2 gene. Next, cells were extracted to be transfected with different mimic, inhibitor and siRNAs to further explore how miR-497 acts to I/R. Western blot analysis and reverse transcription quantitative polymerase chain reaction (RT-qPCR) were conducted to measure expressions of miR-497, Mfn2, Fas, Bcl-2, Bax and Caspase-3 in myocardial tissues and cardiomyocytes after transfection. CCK-8 assay and flow cytometry were used to determine proliferation, cell cycle distribution and apoptosis of cardiomyocytes in each group after transfection. RESULTS: Mice with I/R had myocardial dysfunction but before the injection with miR-497 agomir, the impairment was alleviated. Mfn2 was verified as the target gene of miR-497. The inhibition of miR-497 in turn inhibits Mfn2 expressione and cardiomyocyte apoptosis. The overexpression of miR-497 and Mfn2 gene silencing can lead to the promotion of proliferation capability of mice cardiomyocytes in vitro. Overexpressed miR-497 and Mfn2 gene silencing can also facilitate cell cycle entry and inhibit the apoptosis cardiomyocytes of mice in vitro. CONCLUSION: The present study provided strong evidence that miR-497 promotes proliferation and inhibits apoptosis of cardiomyocytes by downregulating the expression of Mfn2 in a mouse model of myocardial I/R injury.

[Effects of combined applications of pig manure and chemical fertilizers on CH4 and N2O emissions and their global warming potentials in paddy fields with double-rice cropping].

A field experiment was carried out to study the effects of combined applications of pig manure and chemical fertilizers on CH4 and N2O emissions, which were measured using the static chamber/gas chromatography method, and their global warming potentials in typical paddy fields with double-rice cropping in Hunan province. The results showed that the combined applications of pig manure and chemical fertilizers did not change the seasonal patterns of CH4 and N2O emissions from paddy soils, but significantly changed the magnitudes of CH4 and N2O fluxes in rice growing seasons as compared with sole application of chemical fertilizers. During the two rice growing seasons, the cumulative CH4 emissions for the pig manure and chemical nitrogen (N) fertilizer each contributing to 50% of the total applied N (1/2N + PM) treatment were higher than those for the treatments of no N fertilizer (ON), half amount of chemical N fertilizer (1/2N) and 100% chemical N fertilizer (N) by 54.83%, 33.85% and 43.30%, respectively (P < 0.05), whilst the cumulative N2O emissions for the 1/2N + PM treatment were decreased by 67.50% compared with N treatment, but increased by 129.43% and 119.23% compared with ON and 1/2N treatments, respectively (P < 0.05). CH4 was the dominant contributor to the global warming potential (GWP) in both rice growing seasons, which contributed more than 99% to the integrated GWP of CH4 and N2O emissions for all the four treatments. Both GWP and yield-scaled GWP for the treatment of 1/2N + PM were significantly higher than the other three treatments. The yield-scaled GWP for the treatment of 1/2N + PM was higher than those for the N, 1/2N and ON treatments by 58.21%, 26.82% and 20. 63%, respectively. Therefore, combined applications of pig manure and chemical fertilizers in paddy fields would increase the GWP of CH4 and N2O emissions during rice growing seasons and this effect should be considered in regional greenhouse gases emissions inventory.