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A pot experiment was conducted to investigate the growth response, concentration, and transportation characteristics of Cd and Pb in Sedum aizoon L. in four typical Cd/Pb-contaminated soils from northern and southern China with the addition of different levels of water-soluble chitosan （WSC）. The effect of WSC on physicochemical properties, Cd and Pb forms, and functional groups in contaminated soils was determined to clarify the internal mechanism of WSC promoting Cd and Pb accumulation by S. aizoon . The results showed that WSC improved the physicochemical properties of rhizosphere soil by increasing alkaline nitrogen and organic matter contents and significantly reducing pH values. In addition, WSC significantly increased the acid-extractable Cd and Pb in contaminated soils by 16.3%-34.5% and 88.9%-272.4%, respectively. Fourier transform infrared spectra analysis showed that amino （-NH 2 ） and hydroxyl （-OH） groups in soils were significantly increased by adding WSC. Therefore, the coordination of Cd/Pb with -NH 2 and -OH may play important roles in forming complexes and mobilizing Cd/Pb in soils. Moreover, the improvement in the rhizosphere environment of contaminated soils by WSC contributed to promoting Cd and Pb accumulation by S. aizoon . WSC significantly increased the total Cd and Pb accumulation by 14.5%-72.1% and 55.0%-128.5%, respectively, without obvious growth inhibition. Therefore, it is feasible to apply WSC to promote Cd/Pb accumulation using S. aizoon , while the matching of WSC addition amount, soil pollution condition, and plant materials should be further considered in practical application.

Background: Transient receptor potential melastatin 4 (TRPM4) affects immune responses by regulating calcium homeostasis, but its role in calcific aortic valve inflammation remains unclear. This study aimed to assess the expression and function of TRPM4 in patients with or without calcific aortic valve disease (CAVD)., Methods: The mRNA and protein expression levels of TRPM4 and related factors in calcified and noncalcified tissues were measured using qRT-PCR and Western blot. The proteins interacting with TRPM4 were confirmed by RNA pull-down and RNA immunoprecipitation assays. Dual-Luciferase Reporter Assay was performed to confirm the m6A site of TRPM4., Results: The mRNA expression levels of TRPM4, TLR4, IL-6, MCP-1, TNF-α, and NF-κB p65 were significantly higher in calcified aortic valve tissues than in noncalcified tissues, and TRPM4 was significantly positively correlated with inflammation-related factors. The protein expression level of TRPM4, TLR4 and NF-κB p65 were significantly higher in calcified aortic valve tissues than in noncalcified tissues. N6-methyladenosine (m6A) modification of TRPM4 mRNA by METTL3-YTHDF1 up-regulated its expression in CAVD. And TRPM4 promoted the level of inflammation via activation of the JNK-MAPK signaling pathway, after knockdown TRPM4, the production of proinflammatory cytokines was significantly suppressed., Conclusion: The results indicate the pivotal role of TRPM4 in CAVD and highlight METTL3-mediated m6A modification of TRPM4 in promoting inflammation through JNK-MAPK signaling pathway. This work provides potential therapeutic strategy to impede inflammation in CAVD., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors. Published by Elsevier Ltd.)

A 20-day sludge biodrying process was coupled with photocatalysis to improve biodrying efficiency and investigate the effect of photocatalysis on biodegradation. After biodrying, the moisture content in the coupled photocatalytic group (T CA ) and the control group (T UCA ) decreased from 63.61% to 50.82% and 52.94%, respectively, and the volatile solids content decreased from 73.18% to 63.42% and 64.39%, respectively. Neutral proteinase activity decreased by 9.38% and 28.69%, and lipase activity decreased by 6.12% and 26.17%, respectively, indicating that photocatalysis helped maintain neutral proteinase and lipase activities. The Chao1 and Shannon indices showed that photocatalysis increased fungal diversity and reduced bacterial richness and diversity. The β diversity clustering analysis indicated that the bacterial community structure during the thermophilic phase in T CA differed from that in T UCA . The Kyoto Encyclopedia of Genes and Genomes annotation showed that photocatalysis has the potential to promote the synthesis and degradation of ketone bodies. Biodrying coupled with photocatalysis can improve the dewatering of sludge without negatively affecting biodegradation., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Context: Kazinol B (KB), an isoprenylated flavan derived from Broussonetia kazinoki Sieb. (Moraceae) root, has long been used in folk medicine., Objective: This study examines the protective effects of KB and its underlying mechanisms in hypoxia and reoxygenation (H/R)-induced cardiac injury in H9c2 rat cardiac myoblasts., Materials and Methods: H9c2 cells were incubated with various concentrations of KB (0, 0.3, 1, 3, 10 and 30 μM) for 2 h and then subjected to H/R insults. The protective effects of KB and its underlying mechanisms were explored., Results: KB significantly elevated cell viability (1 μM, 1.21-fold; 3 μM, 1.36-fold, and 10 μM, 1.47-fold) and suppressed LDH release (1 μM, 0.77-fold; 3 μM, 0.68-fold, and 10 μM, 0.59-fold) in H/R-induced H9c2 cells. Further, 10 μM KB blocked apoptotic cascades, as shown by the Annexin-V/PI (0.41-fold), DNA fragmentation (0.51-fold), caspase-3 (0.52-fold), PARP activation (0.27-fold) and Bax/Bcl-2 expression (0.28-fold) assays. KB (10 μM) downregulated reactive oxygen species production (0.51-fold) and lipid peroxidation (0.48-fold); it upregulated the activities of GSH-Px (2.08-fold) and SOD (1.72-fold). KB (10 μM) induced Nrf2 nuclear accumulation (1.94-fold) and increased ARE promoter activity (2.15-fold), HO-1 expression (3.07-fold), AKT (3.07-fold) and AMPK (3.07-fold) phosphorylation. Nrf2 knockdown via using Nrf2 siRNA abrogated KB-mediated protective effects against H/R insults. Moreover, pharmacological inhibitors of AKT and AMPK also abrogated KB-induced Nrf2 activation and its protective function., Discussion and Conclusions: KB prevented H/R-induced cardiomyocyte injury via modulating the AKT and AMPK-mediated Nrf2 induction. KB might be a promising drug candidate for managing ischemic cardiac disorders.

The efficiency of sludge dewatering is limited by extracellular polymeric substances (EPS) during biodrying. This study investigated the effect of photocatalysis-mediated EPS degradation on sludge dewatering performance during the sludge biodrying process. The photocatalysis of municipal sludge was first carried out to choose a cost-efficient catalyst. Then sludge biodrying tests were performed using TiO 2 -coated amendment (T CA ) and uncoated amendment (T UCA ) as the control. Municipal sludge photocatalysis results showed that using TiO 2 could efficiently degrade carbohydrates and proteins in the EPS within 60 min. After 20-day biodrying, photocatalysis significantly promoted a reduction in the moisture content and EPS by 17.64% and 6.88%, respectively. The surface-enhanced Raman scattering (SERS) intensities of the C-C-O symmetric stretching vibration peak of D-lactose and the C-S stretching vibration peak of cysteine were significantly decreased by approximately 33.19% and 44.76%, respectively, indicating that photocatalysis indeed promoted the reduction of polysaccharides and cysteine in the EPS, especially after the thermophilic phase. The hydrophilic amino acid content decreased by 23.02%, verifying that photocatalysis could improve EPS hydrophobicity. Consequently, municipal sludge biodrying coupled with photocatalysis promotes sludge EPS degradation and enhances sludge dewaterability, improving the efficiency of sludge biodrying., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

The blunt snout bream (Megalobrama amblycephala) is a typical hypoxia-sensitive fish, and hypoxia stress leads to reduced vitality and yield during aquaculture. To explore the specific adaptation mechanism under hypoxia, the blunt snout bream was treated with hypoxia (DO = 2.0 ± 0.1 mg/L) for 24 h, followed by 3 h of recovery. Our results depicted that the gill filament structure of blunt snout bream changed after hypoxia. During hypoxia for 24 h, the gill filament structure was altered, including a more than 80% expansion of the lamellar respiratory surface area and a proportionate apoptosis decrease in interlamellar cell mass (ILCM) volume. Thus, the water-blood diffusion distance was shortened to less than 46%. During hypoxia for 24 h, the activity of ROS in gill tissue increased significantly (p < 0.05), while the mitochondrial membrane potential decreased significantly (p < 0.05). During hypoxia, mRNA expression level of anti-apoptotic gene Bcl-2 in the gills of blunt snout bream decreased significantly (p < 0.05), while the expression of pro-apoptotic gene Bax mRNA increased significantly (p < 0.05). Thus, the ratio of Bax/Bcl-2 mRNA increased in the gills of blunt snout bream to promote the activity of Caspase-3. Together, our results indicated hypoxia-induced apoptosis in the gills of blunt snout bream through the mitochondrial pathway. In addition, a decreased expression of Phd1 and an increased expression of Hif-1α in gills under hypoxia stress indicates that blunt snout bream may cope with hypoxia-induced apoptosis by enhancing the HIF pathway. These results provide new insights into fish's adaptation strategies and mechanisms of hypoxia., (© 2023. The Author(s), under exclusive licence to Springer Nature B.V.)

With the gradual decrease in freshwater resources, the available space for freshwater aquaculture is diminishing. As a result, saline-alkaline water aquaculture has emerged as a crucial method to fulfill the increasing demand. This study investigates the impact of alkaline water on the growth performance, tissues (gill, liver, and kidney), digestive enzyme activity, and intestinal microbiology in grass carp (Ctenopharyngodon idella). The aquarium conditions were set with sodium bicarbonate (18 mmol/L (LAW), 32 mmol/L (HAW)) to simulate the alkaline water environment. A freshwater group was the control (FW). The experimental fish were cultured for 60 days. The findings revealed that NaHCO 3 alkaline stress significantly reduced growth performance, caused alterations in the structural morphology of gill lamellae, liver, and kidney tissues, and led to decreased activity of intestinal trypsin and lipase amylase (P < 0.05). Analysis of 16S rRNA sequences demonstrated that alkalinity influenced the abundance of dominant bacterial phyla and genera. Proteobacteria showed a significant decrease under alkaline conditions, while Firmicutes exhibited a significant increase (P < 0.05). Furthermore, alkalinity conditions significantly reduced the abundance of bacteria involved in protein, amino acid, and carbohydrate metabolism, cell transport, cell decomposition, and environmental information processing. Conversely, the abundance of bacteria associated with lipid metabolism, energy metabolism, organic systems, and disease functional flora increased significantly under alkalinity conditions (P < 0.05). In conclusion, this comprehensive study indicates that alkalinity stress adversely affected the growth performance of juvenile grass carp, likely due to tissue damage, reduced activity of intestinal digestive enzymes, and alterations in intestinal microorganisms., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

A new hypoxia-tolerant variety of blunt snout bream was obtained by successive breeding of the wild population, which markedly improved hypoxia tolerance. In this study, the hypoxia-tolerant variety was exposed to hypoxia (2.0 mg O 2 ·L -1 ) for 4, 7 days. The contents of blood biochemical indicators including the number of red blood cells (RBC), total cholesterol (T-CHO), total protein (TP), triglyceride (TG), glucose (GLU), and lactic acid (LD) increased significantly (P < 0.05) under hypoxia. The glycogen content in the liver and muscle decreased significantly (P < 0.05) and the LD content in the brain, muscle and liver increased significantly (P < 0.05) under hypoxia. The levels of oxidative stress-related indicators i.e., superoxide dismutase (SOD), malondialdehyde (MDA), glutathione (GSH), catalase (CAT), and total antioxidant capacity (T-AOC) also changed significantly (P < 0.05) in the heart, liver, and intestine of the new variety under hypoxia. Additionally, hypoxia has caused injuries to the heart, liver, and intestine, but it shows amazing repair ability during reoxygenation. The apoptotic cells and apoptosis rate in the heart, liver, and intestine increased under hypoxia. Under hypoxia, the expression of the B-cell lymphomas 2 (Bcl-2) gene in the heart, liver, and intestine was significantly (P < 0.05) down-regulated, while the expression of the BCL2-associated agonist of cell death (Bad) gene was significantly (P < 0.05) up-regulated. These results are of great significance for enriching the basic data of blunt snout bream new variety in response to hypoxia and promoting the healthy development of its culture industry., Competing Interests: Declaration of competing interest The authors declare that they have no competing interests., (Copyright © 2022. Published by Elsevier Inc.)

Blunt snout bream (Megalobrama amblycephala) is sensitive to hypoxia environment. Hypoxia-inducible factor (HIF) is the most critical factor in the HIF pathway, which strictly regulates the hypoxia stress process of fish. In this study, we found six hifα genes in blunt snout bream that demonstrated different expressions under hypoxia conditions. In HEK293T cells, all six hifαs were detected to activate the HRE region by luciferase reporter assay. More importantly, we identified two linkage-disequilibrium SNP sites at exon 203 and 752 of the hif2αb gene in blunt snout bream. Haplotype II (A 203 A 752 ) and its homozygous diplotype II (A 203 A 203 A 752 A 752 ) appeared frequently in a selected strain of blunt snout bream with hypoxia tolerance. Diplotype II has a lower oxygen tension threshold for loss of equilibrium (LOE crit ) over a similar range of temperatures. Moreover, its erythrocyte number increased significantly (p < 0.05) than those in diplotype I and diplotype III strains at 48 h of hypoxia. The enzymes related with hypoxia tolerant traits, i.e., reduced glutathione, superoxide dismutase, and catalase, were also significantly (p < 0.05) induced in diplotype II than in diplotype I or III. In addition, the expression of epo in the liver of diplotype II was significantly (p < 0.01) higher than that in the diplotype I or III strains at 48 h of hypoxia. Taken together, our results found that the hypoxia-tolerant-related diplotype II of hif2αb has the potential to be used as a molecular marker in future genetic breeding of hypoxia-tolerant strain., (© 2023. The Author(s), under exclusive licence to Springer Nature B.V.)

Intercropping a Cd-accumulator with economically valuable crops is common in slightly or moderately Cd-polluted farmland soils. A field experiment was conducted to evaluate the effects of water-soluble chitosan (WSC) on the growth and Cd uptake of the Cd-accumulator Hylotelephium spectabile and soybean ( Glycine max ) during a co-cultivation in Cd-contaminated agricultural soil (WSC, 0 and 10 g·m -2 ). The results indicated that soybean yields were highest in response to the intercropping and WSC treatment. The results from the field trials generally showed that intercropping and WSC treatments significantly decreased Cd concentrations in inedible parts of soybean by 42.9-72.1% (except for stems), in the meantime, increased 95.8%-334.6% in shoot and root tissues of H. spectabile compared with the control ( p  < 0.05). The data revealed that Cd uptake was highest for H. spectabile during the intercropping and WSC treatment. The application of WSC in the intercropping system significantly increased the uptake of Cd by H. spectabile , but not by soybean. The findings of this study suggest that combining an intercropping system with a WSC treatment may be better for remediating Cd-contaminated soils than other methods involving the growth of a single hyperaccumulator.

Biodrying is a treatment to remove moisture using bio-heat generated during organic degradation. Organic matter degradation and microbial metabolism were studied during the whole kitchen waste biodrying, using metagenomic analysis. After the 25-day biodrying process, carbohydrate, protein and lipid contents decreased by 83.7%, 27.8% and 79.3%, respectively, and their degradation efficiencies increased after the thermophilic phase. Lipase activity exceeded 10 mmol d -1 g -1 throughout biodrying. Cellulase and lipase activities recovered by 2.21% and 5.77%, respectively, after the thermophilic phase, while the protease activity had a maximum increment of 347%. Metabolic analysis revealed that carbohydrate, amino acid and lipid metabolism was possibly inhibited by the high temperature, but the relative abundances of related predicted functions recovered by more than 0.9%, 7% and 11%, respectively, by the end of biodrying. Protein function prediction suggests that β-oxidation, fatty acid biosynthesis, and the degradation of cellulose and chitin were possibly enhanced during the thermophilic phase. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis revealed that leucine, isoleucine and lysine could ultimately degraded to acetyl-CoA. Weissella, Aeribacillus and Bacillus were the genera with the most enriched functional genes during the whole biodrying process. These findings help elucidate the microbial degradation processes during biodrying, which provides further scientific support for improving the application of biodrying products., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

In order to explore the distribution characteristics of heavy metal contamination of farmland soil surrounding Luancheng town, Shijiazhuang City, Henan province, the concentrations of As, Cd, Cr, Cu, Hg, Ni, Pb, and Zn in the surface soil and crops were determined and assessed. The principal components were also analyzed for source apportionment. The heavy metal concentrations in crops were further detected, and the non-carcinogenic health risks in the study area were evaluated using the probabilistic risk assessment method, as to provide a theoretical basis for the treatment, prevention, and control of heavy metal pollution in farmland soil in Luancheng. According to the results, ω (Cd), ω (Cr), ω (Cu), ω (Pb), and ω (Zn) in the soils were 0.06-1.08, 22.14-473.47, 12.83-150.74, 10.75-577.72, and 62.23-652.78 mg·kg -1 , which exceeded the standard with over-standard rates reaching 1.83%, 1.22%, 0.61%, 0.61%, and 1.22%, respectively. Further, Cd and Pb were transported into crops, in which Cd concentrations exceeded the standard in some corn samples, and Cd and Pb concentrations exceeded the standard in some wheat samples. The total non-carcinogenic health risks (TTHQ) to the human body caused by the consumption of heavy metals in corn grown in the study area were all less than 1, with no obvious negative effects, and TTHQ was higher than 1 in wheat, increasing the likelihood of negative impacts on the human body. With the influence of the distribution of pollution-related enterprises in the industrial zone, heavy metal concentrations were higher in the south, west, and middle directions of the study area. Among them, the study area soil was slightly contaminated by Cd (Level 1). Cd and Hg had a slight potential ecological risk (Level 2), whereas other heavy metals had low potential ecological risk (Level 1). In general, most of the surface cultivated soil was not obviously polluted by heavy metals in the study area. According to the PMF results and survey, we speculated that soil heavy metals mainly came from soil parent material (52.05%), artificial pollution sources (historical sewage irrigation and industrial manufacture) (32.98%), and atmospheric deposition (14.97%). To summarize, the study area should be divided into a priority protection category and safe utilization category. The input of pollution sources should be strictly controlled for the priority protection category, and alternative planting, rotating, and fallow should be implemented for the safe utilization category to reduce the risk of standard-exceeding agricultural products.

Background: The grass carp has great economic value and occupies an important evolutionary position. Genomic information regarding this species could help better understand its rapid growth rate as well as its unique body plan and environmental adaptation., Results: We assembled the chromosome-level grass carp genome using the PacBio sequencing and chromosome structure capture technique. The final genome assembly has a total length of 893.2 Mb with a contig N50 of 19.3 Mb and a scaffold N50 of 35.7 Mb. About 99.85% of the assembled contigs were anchored into 24 chromosomes. Based on the prediction, this genome contained 30,342 protein-coding genes and 43.26% repetitive sequences. Furthermore, we determined that the large genome size can be attributed to the DNA-mediated transposable elements which accounted for 58.9% of the repetitive sequences in grass carp. We identified that the grass carp has only 24 pairs of chromosomes due to the fusion of two ancestral chromosomes. Enrichment analyses of significantly expanded and positively selected genes reflected evolutionary adaptation of grass carp to the feeding habits. We also detected the loss of conserved non-coding regulatory elements associated with the development of the immune system, nervous system, and digestive system, which may be critical for grass carp herbivorous traits., Conclusions: The high-quality reference genome reported here provides a valuable resource for the genetic improvement and molecular-guided breeding of the grass carp., (© 2022. The Author(s).)

Background: Taxifolin is a natural dihydroflavonol found in many plants and health products. In the present study, its anti-obesity and gut microbiota modulating effects were studied. C57BL/6J mice were fed with a high-fat diet (HFD) supplemented with taxifolin (0, 0.5 and 1 mg mL -1 , respectively) in drinking water for 15 weeks., Results: Taxifolin supplementation showed no influence on food and water intake. However, it decreased body weight gain, inhibited fat accumulation, and decreased total cholesterol and triacylglycerol level in mice liver. Taxifolin enhanced superoxide dismutase (SOD) activity in mice liver, which in turn protected the liver from lipid peroxidation damage. It also improved insulin resistance in obese mice. Metagenomic analysis of bacterial 16S rRNA demonstrated that HFD decreased gut microbiota diversity and caused dysbiosis. However, taxifolin improved the gut microbiota diversity and decreased the Firmicutes/Bacteroidetes ratio. In particular, it inhibited Proteobacteria from blooming, this being a signature of dysbiosis in gut microbiota., Conclusion: Taxifolin ameliorated the symptoms of obesity, hepatic steatosis, lipid peroxidation, insulin resistance, and gut microbiota dysbiosis in HFD fed C57BL/6J mice. © 2021 Society of Chemical Industry., (© 2021 Society of Chemical Industry.)

Background: Blunt snout bream (Megalobrama amblycephala) is sensitive to hypoxia. A new blunt snout bream strain, "Pujiang No.2", was developed to overcome this shortcoming. As a proteasome inhibitor, bortezomib (PS-341) has been shown to affect the adaptation of cells to a hypoxic environment. In the present study, bortezomib was used to explore the hypoxia adaptation mechanism of "Pujiang No.2". We examined how acute hypoxia alone (hypoxia-treated, HN: 1.0 mg·L - 1 ), and in combination with bortezomib (hypoxia-bortezomib-treated, HB: Use 1 mg bortezomib for 1 kg fish), impacted the hepatic ultrastructure and transcriptome expression compared to control fish (normoxia-treated, NN)., Results: Hypoxia tolerance was significantly decreased in the bortezomib-treated group (LOE crit , loss of equilibrium, 1.11 mg·L - 1 and 1.32 mg·L - 1 ) compared to the control group (LOE crit , 0.73 mg·L - 1 and 0.85 mg·L - 1 ). The HB group had more severe liver injury than the HN group. Specifically, the activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in the HB group (52.16 U/gprot, 32 U/gprot) were significantly (p < 0.01) higher than those in the HN group (32.85 U/gprot, 21. 68 U/gprot). In addition, more severe liver damage such as vacuoles, nuclear atrophy, and nuclear lysis were observed in the HB group. RNA-seq was performed on livers from the HN, HB and NN groups. KEGG pathway analysis disclosed that many DEGs (differently expressed genes) were enriched in the HIF-1, FOXO, MAPK, PI3K-Akt and AMPK signaling pathway and their downstream., Conclusion: We explored the adaptation mechanism of "Pujiang No.2" to hypoxia stress by using bortezomib, and combined with transcriptome analysis, accurately captured the genes related to hypoxia tolerance advantage., (© 2022. The Author(s).)

Scope: Alzheimer's disease (AD) is a neurodegenerative disease with phenomena of cognitive impairments. Oxidative stress and cholinergic system dysfunction are two widely studied pathogenesis of AD. Dihydromyricetin (DMY) is a natural dihydroflavonol with many bioactivities. In this study, it is aimed to investigate the effects of DMY on cognitive impairment in d-galactose (d-gal) induced aging mice., Methods and Results: Mice are intraperitoneally injected with d-gal for 16 weeks, and DMY is supplemented in drinking water. The results show that DMY significantly improves d-gal-induced cognitive impairments in novel object recognition and Y-maze studies. H&E and TUNEL staining show that DMY could improve histopathological changes and cell apoptosis in mice brain. DMY effectively induces the activities of catalase, superoxide dismutase and glutathione peroxidase, and reduces malondialdehyde level in mice brain and liver. Furthermore, DMY reduces cholinergic injury by inhibiting the activity of Acetylcholinesterase (AChE) in mice brain. In vitro studies show that DMY is a non-competitive inhibitor of AChE with IC50 value of 161.2 µg mL -1 ., Conclusion: DMY alleviates the cognitive impairments in d-gal-induced aging mice partly through regulating oxidative stress and inhibition of acetylcholinesterase., (© 2021 Wiley-VCH GmbH.)

Blunt snout bream plays an important role in freshwater aquaculture in China, but the development of its culture industry has been restricted by increasing hypoxia problem. Through the breeding of wild blunt snout bream populations (F0), a hypoxia-tolerant new variety (F6) was obtained. In this study, the new variety was stressed under low oxygen concentration (2.0 mg·L -1 ) for 4 and 7 days, the morphological structure of the gill tissue showed a striking change, the interlamellar cell mass (ILCM) volume reduced significantly (P < 0.05), and the lamellar respiratory surface area enlarged significantly (P < 0.05), compared to normoxic controls. After 7 days of oxygen recovery, gill remodeling was completely reversed. Additionally, the TUNEL-positive apoptotic fluorescence signals increased in the gills exposed to hypoxia up to 4 and 7 days; the apoptosis rate also increased significantly (P < 0.05). Under 4 and 7 days of hypoxia stress, the expression of anti-apoptotic gene Bcl-2 in the gills downregulated significantly (P < 0.05), with the significantly (P < 0.05) upregulated expression of pro-apoptotic gene Bad. Furthermore, under hypoxia stress, the activity or content of oxidative stress-related enzymes (superoxide dismutase (SOD), catalase (CAT), malondialdehyde (MDA), and glutathione (GSH)) in gill tissue increased to varying degrees compared to normoxic controls. These results offer a new perspective into the cellular and molecular mechanism of hypoxia-induced gill remodeling in blunt snout bream and a theoretical basis for its hypoxia adaptation mechanism., (© 2022. The Author(s), under exclusive licence to Springer Nature B.V.)

N-ethyl-N-nitrosourea (ENU) selection is a useful technique to generate new mutations that may cause some functional changes in the gene. Through our previous genomic bulked segregant analysis (BSA), one single nucleotide polymorphism (SNP) at the 3' UTR of Toll interacting protein gene (TOLLIP 982T>C ) was identified in grass carp (Ctenopharyngodon idella) subjected to ENU-induced mutagenesis. We found that the overexpression of cid-miR-nov-1043 mimics significantly suppressed the luciferase activity of the TOLLIP 3' UTR, but TOLLIP 982T>C mutation at the target site can decrease the binding affinity between the miRNA cid-miR-nov-1043 and TOLLIP 3' UTR, reducing the inhibition of TOLLIP mRNA transcription in grass carp subjected to ENU-induced mutagenesis. More importantly, we demonstrated that TOLLIP mRNA transcription levels in the gills, liver, kidney and the isolate white cells of the mutant grass carp were significantly (p < 0.01) higher than those in the corresponding tissues from the wild-type grass carp following infection with Grass Carp Reovirus (GCRV) for seven days, while the downstream gene of TOLLIP transforming growth factor β-activated kinase 1 (TAK1) and TAK1-binding protein 1 (TAB1), were higher expressed in wild-type grass carp. As a negative regulator in the pro-inflammatory pathway of NF-κB, TOLLIP inhibits the excessive inflammation in ENU grass carp after GCRV infection. Consistent with the TOLLIP expression, histopathological results demonstrated more severe inflammation in wild-type grass carp, compared to the TOLLIP 982T>C mutant grass carp on the seventh day. Severe inflammation will lead to thoroughly infiltration of chloride and inflammatory cells in the gill filaments. This seriously hindered the exchange of oxygen, which ultimately disrupted blood circulation. Meanwhile, the survival rate of the mutant grass carp was significantly (p < 0.01) higher than that of the wild-type grass carp, indicating that the TOLLIP 982T>C mutants showed strong anti-viral abilities. Our results revealed that an SNP in the TOLLIP 3' UTR may contribute to the suppression of serve inflammation subjected to ENU-induced mutagenesis following GCRV infection, which may be helpful for future resistant breeding development of grass carp., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

MiR-6870-3p acts as a crucial regulator of gene expression at the posttranscriptional level and participates in immune responses. However, the roles of miR-6870-3p and its target genes and their underlying mechanisms in the inflammatory responses of epicardial adipose tissues (EATs) are unknown.MiRNA microarray was used to collect the miRNA expression profiles of EATs from five patients with coronary artery disease (CAD) and four individuals without CAD (n-CAD). Quantitative real-time polymerase chain reaction (qRT-PCR) was applied to examine the expression of miR-6870-3p in the EATs. The mRNA and protein expression levels of Tollip and the key genes of the Toll-like receptor 4 (TLR4) signaling pathway were examined by qRT-PCR and Western blot analysis. The levels of inflammatory factors in the cell supernatant were measured by enzyme-linked immunosorbent assay (ELISA). We used a dual-luciferase reporter assay to validate the target gene of miR-6870-3p. The protein expression levels of c-Jun N-terminal kinase (JNK) and nuclear factor kappa B (NF-κB) were measured by Western blot analysis.Our results showed that miR-6870-3p was higher in the CAD EATs than in the n-CAD EATs. MiR-6870-3p was positively correlated with TLR4, interleukin (IL)-6, JNK, NF-κB (p65), and tumor necrosis factor (TNF)-α in the CAD EAT samples. Lipopolysaccharide (LPS) treatment upregulated the miR-6870-3p expression and downregulated the Tollip expression in the macrophages. When the macrophages were stimulated with LPS, MiR-6870-3p upregulation also aggravated the production of proinflammatory cytokines. The result of the luciferase reporter assays confirmed that miR-6870-3p directly targets Tollip. Moreover, miR-6870-3p upregulation in the macrophages resulted in the activation of the JNK/NF-κB pathway.Our study showed that miR-6870-3p regulates human EAT inflammation by targeting the Tollip-mediated JNK and NF-κB signaling pathways.

Three shell materials, lecithin (ZNP-L), chitosan (ZNP-CH) and sodium caseinate (ZNP-SC), were used to prepare core-shell zein nanoparticles. Astilbin was encapsulated as a model flavonoid to compare the influence of the shell materials on zein nanoparticles both in vitro and in vivo . The particle size was moderately increased by lecithin and sodium caseinate, but notably increased by chitosan. All the shell materials provided good redispersibility for the nanoparticles and significantly improved the colloidal stability. Chitosan and sodium caseinate significantly delayed and decreased the feces excretion of astilbin in rats, while lecithin exhibited a very weak effect. The results may be attributed to the difference in mucoadhesive properties between the shell materials. As a consequence, the bioavailability values of astilbin in rats were 18.2, 9.3 and 1.89 times increased through ZNP-CH, ZNP-SC and ZNP-L compared with that of free astilbin, respectively.

Background: N-ethyl-N-nitrosourea (ENU) mutagenesis is a useful method for the genetic engineering of plants, and the production of functional mutants in animal models including mice and zebrafish. Grass carp reovirus (GCRV) is a haemorrhagic disease of grass carp which has caused noteworthy losses in fingerlings over the last few years. To overcome this problem, we used ENU mutant grass carp in an attempt to identify functional resistance genes for future hereditary rearing projects in grass carp., Results: This study used ENU-mutated grass carp to identify genetic markers associated with resistance to the haemorrhagic disease caused by GCRV. Bulked segregant analysis (BSA) was performed on two homozygous gynogenetic ENU grass carp groups who were susceptible or resistant to GCRV. This analysis identified 466,162 SNPs and 197,644 InDels within the genomes of these mixed pools with a total of 170 genes annotated in the associated region, including 49 genes with non-synonymous mutations at SNP sites and 25 genes with frame shift mutations at InDel sites. Of these 170 mutated genes, 5 randomly selected immune-related genes were shown to be more strongly expressed in the resistant group as compared to the susceptible animals. In addition, we found that one immune-related gene, EPHB2, presented with two heterozygous SNP mutations which altered the animal's responded to GCRV disease. These SNPs were found in the intron region of EPHB2 at positions 5859 (5859 G > A ) and 5968 (5968 G > A ) and were significantly (p = 0.002, 0.003) associated with resistance to GCRV. These SNP sites were also shown to correlate with the GCRV-resistant phenotype in these ENU grass carp. We also evaluated the mortality of the different ENU fish genotypes in response to GCRV and the SNPs in EPHB2. The outcomes of these evaluations will be useful in future selections of GCRV-resistant genes for genetic breeding in grass carp., Conclusion: Our results provide a proof of concept for the application of BSA-sequence analysis in detecting genes responsible for specific functional genotypes and may help to develop better methods for marker-assisted selection, especially for disease resistance in response to GCRV.

This research investigated a synthetic amendment to improve composting and resource recycling of pig manure and biogas residue. We further examined whether adding a synthetic amendment impacts the microbial ecosystem in the composted materials. Three mixing ratios were used to investigate composting performance: no synthetic amendment (T0), 5% synthetic amendment (T1), and 10% synthetic amendment (T2) (T1 and T2 were measured as a wet weight ratio). There were no significant differences in the fundamental characteristics between composting products in T0 and T1. The moisture content of composting material in T0, T1, and T2 significantly decreased from a baseline of approximately 65% to 35.5%, 37.3%, and 55.9%, respectively. Meanwhile, the germination index significantly increased to 111.6%, 155.6%, and 62.3%, respectively. When an optimal proportion of synthetic amendment was added, T1 showed high degree of humification, lignocellulase activities, and effective biodegradation. Firmicutes, Actinobacteria, Proteobacteria, and Bacteroidetes were the dominant bacteria, while Ascomycota and Basidiomycota were the dominant fungi in all treatment groups. Amino sugar and nucleotide sugar metabolism, glycolysis, starch, and sucrose metabolism were among the primary pathways in predicted functions. The synthetic amendment can generate a mature composting product and can be reused or recycled to conserve resources., Competing Interests: Declaration of Competing Interest The author declare that there is no conflict of interest., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Aim: Epigallocatechin-3-gallate (EGCG), the major polyphenol found in green tea, exerts multiple protective effects against cardiovascular diseases, including cardiac hypertrophy. However, the molecular mechanism underlying its anti-hypertrophic effect has not been clarified. This study revealed that EGCG could inhibit pressure overload-induced cardiac hypertrophy by regulating the PSMB5/Nmnat2/SIRT6-dependent signalling pathway., Methods: Quantitative real-time polymerase chain reaction and western blotting were used to determine the expression of mRNA and protein respectively. A fluorometric assay kit was used to determine the activity of SIRT6, a histone deacetylase. Luciferase reporter gene assay and electrophoretic mobility shift assay were employed to measure transcriptional activity and DNA binding activity respectively., Results: EGCG could significantly increase Nmnat2 protein expression and enzyme activity in cultured neonatal rat cardiomyocytes stimulated with angiotensin II (Ang II) and heart tissues from rats subjected to abdominal aortic constriction. Nmnat2 knockdown by RNA interference attenuated the inhibitory effect of EGCG on cardiac hypertrophy. EGCG blocked NF-κB DNA binding activity induced by Ang II, which was dependent on Nmnat2 and the subsequent SIRT6 activation. Moreover the activation of PSMB5 (20S proteasome subunit β-5, chymotrypsin-like) was required for EGCG-induced Nmnat2 protein expression. Additionally, we demonstrated that EGCG might interact with PSMB5 and inhibit the activation of the proteasome., Conclusions: These findings serve as the first evidence that the effect of EGCG against cardiac hypertrophy may be, at least partially, attributed to the modulation of the PSMB5/Nmnat2-dependent signalling pathway, suggesting the therapeutic potential of EGCG in the prevention and treatment of cardiac hypertrophy., (© 2020 Scandinavian Physiological Society. Published by John Wiley & Sons Ltd.)

Biodrying in conjunction with compound stone amendment was used to treat kitchen waste, which improved biodrying. After 16 days, the pile moisture content decreased from 68.8% to 23.0%. Lignin, cellulose and hemicellulose concentrations decreased from 104.6 mg g -1 d.b., 322.9 mg g -1 d.b. and 155.9 mg g -1 d.b., respectively, to 74.0 mg g -1 d.b., 224.8 mg g -1 d.b. and 134.5 mg g -1 d.b., respectively. The Shannon index for bacteria increased from 2.5 to 3.1, while for fungi, it decreased from 4.6 to 0.6. The relative abundances of Amino Acid Metabolism and Carbohydrate Metabolism exceeded 7%. The thermophilic phase during the process inactivated the pathogenic microorganisms, increased the bacterial diversity, decreased the fungal diversity, and potentially improved the metabolism of nutrients, including amino acids, carbohydrates, lipids and vitamins. The biomarker analysis and predicated protein sequences provide genetic evidence to elucidate why the thermophilic phase is the peak time for nutrient metabolism., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

The purpose of this study is to explore the expression of miRNA-590-5p, an exosome of gastric cancer (GC), and to evaluate the suitability of miR-590-5p, an exosome with its own clinical characteristics. Serum samples from 168 gastric cancer patients and 50 matched controls were collected and exosomal RNAs were extracted. After that, miR-590-5p is analyzed by quantitative polymerase chain reaction (qRT-PCR), which is more related to clinical and pathological parameters and patient monitoring data. MGC-803 and HGC-27 cells were treated by miR-590-5p mimics, and then the changes of cell fluidity and invasiveness were monitored. The results showed that the expression level of miR-590-5p in exosomes of healthy observation group, early (I and II) stage group, and late stage (III) group was 30.34 ± 6.35, 6.19 ± 0.81, and 2.9 ± 0.19, respectively (all p < 0.05). ROC (receiver-operating characteristic curve) showed that the AUC (area under the curve) of exosomal miR-590-5p was 0.810 with 63.7% sensitivity and 86% specificity. The expression of exosomal miR-590-5p in serum was related to clinical stage ( p = 0.008), infiltration depth, and the expression level of ki-67 ( p < 0.001). In addition, Kaplan-Meier analysis showed that the decrease of explicit level of exosomal miR-590-5p was related to the decrease of overall survival rate ( p < 0.001). Cox regression analysis showed that miR-590-5p can be used as an independent predictor. Furthermore, upregulation of miR-590-5p inhibited cell migration and invasion in MGC-803 cells and HGC-27 cells. The serum expression level of exosomal miR-590-5p may be a biomarker, which is potentially useful and noninvasive for early detection and prediction of GC. In addition, miR-590-5p can play a role in eliminating carcinogens by actively regulating the malignant potential of gastric cancer., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Zheng, Xu, Hu, Lv, Xie, Huang, Zhang, Chen, Fu and Cheng.)

In order to fully explore the spatial distribution of soil heavy metal contaminants in farmland soil surrounding a typical industrial area in Dakuai town, Xinxiang city, Henan Province, the concentrations of As, Cd, Cr, Cu, Ni, Pb, and Zn in the surface soil and within the soil profile were determined and assessed. The principal components were also analyzed for source apportionment to provide a theoretical basis for the control and prevention of heavy metal pollution. According to the results, the soils in the study area are severely contaminated by Cd and Cu and moderated contaminated by As due to the battery manufacture and Cu (e.g., pipe and wire) processing. The concentrations of Cd, Cu, and As in soils were (2.56±1.23) mg·kg -1 , (205.58±157.49) mg·kg -1 , and (15.27±4.14) mg·kg -1 , respectively, which exceeds standards by 100%, 89.44%, and 3.40%, respectively. Accounting for the influence of pollution sources, terrain, runoff erosion, and prevailing wind direction, all heavy metal concentrations were higher in the south direction, lower in the north direction. The concentrations of Cd and Cu in soil profile samples decreased with depth, with highest concentrations at the surface, indicating the strong effect of industrial activities. Arsenic concentrations varied little with soil depth due to its strong migration ability in alkaline soil, again suggesting an industrial source. Among them, the soil of study area was severely contaminated by Cd and Cu (Level 5). Cd poses a severe potential ecological risk (Level 5) and Cu poses a medium risk (Level 2) in the study soils, while other heavy metals show low potential ecological risk (Level 1). The order of the risk identified was Cd > Cu > Ni > As > Pb > Cr > Zn. In addition, the combined potential ecological risk of heavy metals in the target area is severe. The principal component analysis showed that the high As, Cd, and Cu contents are mainly derived from industrial areas. Therefore, it is urgent to remediate Cd and Cu soil contamination in this area and implement precautions to limit As contamination.

Dihydromyricetin (DMY) encapsulated zein-caseinate nanoparticles (DZP) were fabricated by antisolvent method. The encapsulation and loading efficiency of DMY in DZP were 90.2% and 22.6% as determined by HPLC. DZP is spherical with particle size and ζ potential of 206.4 nm and -29.6 mV, respectively. Physicochemical characterization showed that DMY existed in amorphous form in DZP and its interaction with proteins was found. The fabrication of DZP significantly improved the stability of DMY. Besides, the diffusion rate of DMY in DZP was faster than its suspensions in both simulated gastric and intestinal fluid. The adhesion of DMY in mice gastrointestinal tract was also improved. Besides DMY itself, its methylated metabolites with further sulfation and glucuronide were identified in rat plasma by UPLC-QTOF-MS. UPLC-QqQ-MS/MS quantitative analysis showed that the oral bioavailability of DMY was 1.95 times enhanced. Besides, the concentration of DMY metabolites in plasma were all increased., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Finding an economical amendment, available in a steady supply, is needed to support the biodrying industrialization. This research developed a recyclable biodrying amendment (RBA) to condition the biodrying of sewage sludge. The pilot-scale treatment (T R ), which included the addition of equivalent weights of RBA and sawdust as amendments, resulted in a higher pile temperature and longer thermophilic phase compared to the control (T C ), which used only sawdust as an amendment. The final moisture content levels were below 50% with both T R and T C . The heat use efficiency for water evaporation was 72.2% and 73.0% in T R and T C , respectively. The activity of α-amylase and cellulose 1,4-β-cellobiosidase increased during the thermophilic phase, while the activity of endo-1,4-β-glucanase and endo-1,4-β-xylanase decreased during the thermophilic phase with both T R and T C . The fourier-transform infrared spectra indicated that adding the RBA resulted in good biodegradability of the lipids, proteins, and polysaccharides. The humic acid to fulvic acid ratio in T R and T C increased from 0.33 (T R ) and 0.35 (T C ) on day 0-0.46 (T R ) and 0.45 (T C ) on day 21, indicating the humification process. The RBA recovery rate was 95.6% and can be reused. These findings highlight that adding RBA showed satisfactory biodrying performance, reduced the amendment cost, and the biodrying product could be incinerated without energy deficit., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

A field experiment with an orthogonally designed experiment L9(3 4 ) was designed to investigate the effect of different N, P, and K levels on plant growth and Cd uptake by Hylotelephium spectabile . The results showed that the biomass of H. spectabile significantly increased with the N application rate. The highest dry weight in the shoot occurred in the treatments with a high level of N (337.5 kg ·hm -2 ), which was 0.86-2.00 times higher than the value with no fertilizer treatment. The addition of K contributed to promoting the Cd absorption of H. spectabile , while no effect was observed when N and P were added. Consequently, NPK fertilizers contribute to increasing the Cd uptake of H. spectabile , and the N and K fertilizer play important role in plant growth and Cd absorption respectively. Moreover, the effect of fertilizers on Cd uptake of H. spectabile was in the order of N > K > P, which indicated that N fertilizer was the main factor for promoting the Cd phytoextraction efficiency of H. spectabile by increasing the biomass. Therefore, the application of high levels of N combined with moderate levels of P and K will be an effective approach to improve the Cd phytoremediation efficiency of H. spectabile by promoting its growth, and the Cd uptake can be increased by a factor of 0.9-2.2 compared to no fertilizer treatment condition.

A Gram-stain-negative, rod-shaped bacterium, designated XJ4 T , was isolated from oil-contaminated water, collected from Xinjiang Province, north-west PR China (45° 1' 27″ N, 85° 6' 14″ E). Growth occurred at 20-45 °C (optimum, 30 °C) and pH 6.0-10.0 (optimum, pH 6.0-7.0). Strain XJ4 T could tolerate up to 7 % (w/v) NaCl and grow optimally in the absence of NaCl. Phylogenetic analysis based on comparative sequence analysis of 16S rRNA gene sequences indicated that strain XJ4 T belonged to the genus Frigidibacter , and that was closely related to Frigidibacter mobilis cai42 T (97.2 %), Frigidibacter albus SP32 T (97.0 %) and Rhodobacter aestuarii JA296 T (97.0 %). The average nucleotide identity values between XJ4 T and three type strains were 77.9, 77.6 and 71.9 %, respectively. The DNA G+C content of strain XJ4 T was 69.5 mol%. The sole respiratory quinone was Q-10. The major cellular fatty acid was summed feature 8 (C 18 : 1   ω7 c and/or C 18 : 1   ω6 c ), C 18 : 0 and 11-methyl C 18 : 1   ω7 c . The polar lipids were diphosphatidylglycerol, phosphatidylglycerol, unidentified phospholipids, an unidentified aminolipid and unidentified lipids. On the basis of phenotypic, chemotaxonomic and phylogenetic analyses, strain XJ4 T represents a novel species of the genus Frigidibacter , for which the name Frigidibacter oleivorans sp. nov. is proposed. The type strain is XJ4 T (=CGMCC 1.13778 T =LMG 30952 T ).

Accumulating clinical and epidemiological evidence indicates a close relationship between diabetes mellitus and dementia. The ginsenoside compound K (CK) has been reported to ameliorate diabetes mellitus and confer protection to the central nervous system. In this study, we investigated whether CK could improve memory impairment and cognitive dysfunction in diabetic db/db mice. Firstly, we found that CK treatments significantly improved behavioral impairment and cognitive dysfunction based on Morris water maze, Y-maze, and fear conditioning tests. Besides, CK decreased the fasting glucose level, increased lipid metabolism, and ameliorated glucose tolerance, insulin sensitivity, and dyslipidemia in diabetic db/db mice. In addition, CK treatments alleviated oxidative stress and inhibited the inflammatory response in hippocampal tissue. Further investigations showed that CK treatments inhibited the NLRP3 inflammasome pathway, as evidenced by the declined expression of TXNIP, NLRP3 inflammasomes, ASC, cleaved caspase-1, and mature IL-1β in hippocampal tissues. Moreover, CK treatments alleviated ER stress via down-regulating the level of BiP, CHOP, p-PERK, p-IRE1α and ATF6 in the hippocampus of db/db mice. These results suggest that CK improves memory and cognitive dysfunction, possibly by ameliorating glucose tolerance, insulin sensitivity, and dyslipidemia, suppressing oxidative stress and inflammatory response and modulating the NLRP3 inflammasome pathway and ER stress.

Objective: Glioblastoma (GBM) is a common type of cancer with high mortality. Epithelial to mesenchymal transition (EMT) plays a vital role in the development of glioblastoma. The aim of this study is to evaluate the role of miR-125a-5p in glioblastoma and in the tumorigenesis of chemotherapeutic drug-resistant cancer stem-like cells in brain glioma., Methods: The role of miR-125a-5p in the regulation of CSCs, EMT, migration, and invasion in glioblastoma was measured in this study., Results: We showed the roles of miR-125a-5p in the regulation of CSCs, EMT, migration, and invasion in glioblastoma. miR-125a-5p can inhibit the CSCs phenotype and EMT in glioblastoma cells. In addition, its over-expression can significantly regulate CSCs-associated genes and EMT-associated gene expression in glioblastoma cells., Conclusions: We concluded that miR-125a-5p is one of the key microRNAs regulating CSCs and EMT programs in glioblastoma. The results suggested that miR-125a-5p might be a novel therapy target for glioblastoma.

A Gram-stain-negative, non-motile and ovoid bacterial strain, designated 4-2 T , was isolated from oil-contaminated water which was collected from Xinjiang Province, north-west PR China. The 16S rRNA gene sequence analysis showed that strain 4-2 T belonged to the genus Paracoccus . The species with highest similarity to strain 4-2 T was Paracoccus saliphilus YIM 90738 T (97.83 %), followed by ' Paracoccus siganidrum ' M26 (97.83 %) and Paracoccus endophyticus SYSUP0003 T (97.25 %). The average nucleotide identity values between 4-2 T and three type strains were 84.69, 77.88 and 74.07 %, respectively. The genomic DNA G+C content of strain 4-2 T was 61.4 mol%. Chemotaxonomical characteristic results showed that the respiratory quinone was ubiquinone Q-10 and the major fatty acids were summed feature 8 (C 18 : 1 ω 7 c or C 18 : 1 ω 6 c ) and C 19 : 0 cyclo ω 8 c . The polar lipid profile consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylcholine, unidentified phospholipids, an unidentified aminolipid and an unidentified polar lipid. The predominant polyamines were putrescine, cadaverine and spermidine. On the basis of phenotypic, chemotaxonomic and phylogenetic inferences, strain 4-2 T represents a novel species of the genus Paracoccus , for which the name Paracoccus alkanivorans sp. nov. is proposed. The type strain is 4-2 T (=CGMCC 1.13669 T =LMG 30882 T ).

The suppressor of cytokine signaling 3 (SOCS3) negatively regulates the responses of various immune cytokines. In this study, we identified socs3s genes of blunt snout bream. 209- and 216-aa long peptides are encoded by socs3a and socs3b genes, respectively. The socs3s mRNAs are expressed consistently during the entire process of embryonic development. Whole-mount in situ hybridization detected socs3a in the eyes and posterior somites at 12 h post fertilization (hpf), transcribed at the otic vesicle at 24 hpf, and transcribed at the eyes, brain, and otic vesicle at 36 hpf; while the socs3b mRNA was transcribed at the notochord at 12 hpf, expressed in the brain, eyes, and tailbud at 24 hpf, and detected in the brain at 36 hpf. The expression of socs3a is slightly different from that of socs3b in tissues of juvenile and adult blunt snout bream. After recombinant human growth hormone (hGH) treatment, the transcript levels of socs3s of blunt snout bream were increased in gills, spleen, kidney, and gonads. After Aerononas hydrophila infection, the mRNA levels of socs3s of blunt snout bream were significantly increased in the liver, spleen, intestine, and kidney tissues. Blunt snout bream were susceptible to various pathogenic microorganisms, we intraperitoneally injected blunt snout bream with A. hydrophila to explore the immune mechanism of socs3s. These results suggested that socs3s of blunt snout bream plays important roles in the regulation of embryonic development and tissue growth, and that socs3s may also play key roles in regulating the bacterial-induced congenital immune response. Socs3s genes has the potential to be used as targeted genes to improve the immunity against bacteria, which is conducive to the improvement of production and breeding., (Copyright © 2019 Elsevier Inc. All rights reserved.)

The suppressor of cytokine signaling 1 (SOCS1) is an essential feedback regulator extensively involved in many different cytokine signaling pathways, such as regulation of the immune system and growth of organism. However, the molecular and functional information on socs1 genes in freshwater fish is unclear. In the present paper, we identified and characterized the full-length closely related but distinct socs1 genes (socs 1a and -1b) in blunt snout bream (Megalobrama amblycephala). The bioinformatic analysis results showed that duplicated socs1s shared majority conserved motifs with other vertebrates. Both socs1a and -1b mRNAs were detected throughout embryogenesis, and gradually increase and then constantly expressed after 16 hpf. Whole-mount in situ hybridization demonstrated that socs1a and socs1b mRNAs were detected in the brain at 12hpf and 24hpf, and in the notochord and brain at 36hpf. In adult fish, the socs1a mRNA were strongly expressed in the heart, eye, kidney, spleen and gonad, but were found to be relatively low in the intestine and liver. On the other hand, the expression of socs1b mRNA was significantly high in the muscle, eye and spleen, and relatively low in the intestine, liver, skin and heart. The results of hGH treatment experiment showed that socs1a and 1b mRNAs were upregulated markedly in the kidney, muscle and liver. Overexpression of socs1s significantly inhibit the GH and JAK/STAT factor stat3 and the inhibitory effect of SOCS1s on GH may be involved in JAK-STAT signaling pathway. These results indicate that SOCS1 plays an important role in regulating growth and development., (Copyright © 2019 Elsevier Inc. All rights reserved.)

In the present study, a simple and efficient method based on orthogonal experimental design and macroporous resin chromatography was established for the first time for enrichment of polymethoxyflavones (PMFs) from the peels of Citrus reticulata 'Chachi' (CRC). The optimum conditions of extraction and enrichment process were as follows: use of a liquid to solid ratio of 1 to 14, two-hour extraction time repeated twice with 70% ethanol; use of HPD-450 macroporous resin, wash solvent of purified water and 25% aqueous ethanol, and 70% aqueous ethanol as desorption solvent. The purity of PMFs in the resulting extract reached 62.26%. Our data indicate that the PMFs purified could potently alleviate high-fat diet-induced hyperlipidaemia. The PMFs were nontoxic as determined by acute toxicity test. The method established was suitable for large-scale separation of PMFs from CRC peels and the PMFs might be developed as an anti-hyperlipidaemia agent or dietary supplement., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The traditional Chinese medicine Citri Reticulatae Pericarpium (CRP) was mainly originated from the dried pericarp of Citrus reticulata 'Chachi' (Crc), Citrus reticulata 'Dahongpao' (Crd), Citrus reticulata 'Unshiu' (Cru) and Citrus reticulata 'Tangerina' (Crt) in China. Since these four cultivars have great similarities in morphology, reliable methods to differentiate CRP cultivars have rarely been reported. To discriminate the differences of these CRP cultivars, herein an efficient and reliable method by combining metabolomics, DNA barcoding and electronic nose was first established. The hierarchical three-step filtering metabolomics analysis based on liquid chromatography-quadrupole time-of-flight mass spectrometry (LC-QTOF-MS) indicated that 9 species-specific chemical markers including 6 flavanone glycosides and 3 polymethoxyflavones could be considered as marker metabolites for discrimination of the geoherb Crc from other cultivars. A total of 19 single nucleotide polymorphism (SNP) sites were found in nuclear internal transcribed spacer 2 (ITS2) of CRP, and three stable SNP sites (33, 128 and 174) in the ITS2 region can distinguish the four CRP cultivars. The electronic nose coupled with chemometrics could also be used to effectively distinguish Crc from other CRP cultivars. Therefore, our results indicated that the integrated method will be an effective strategy for discrimination of similar herbal medicines., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Two-dimensional gel electrophoresis (2-DE) was combined with liquid chromatography-mass spectrometry (LC-MS/MS) to identify the differential proteomics of grass carp gills after hypoxic stress to better understand the roles of proteins in the hypoxic response and to explore the possible molecular mechanisms. Protein spots were obtained from a hypoxia-stressed group (372 ± 11 individuals) and a control group (406 ± 14 individuals) using the lmage Master 2D Platinum 7.0 analysis software. Fifteen protein spots were expressed differentially in the hypoxia-stressed group and varied significantly after exposure to the hypoxic conditions. In addition, these differential proteins were identified by mass spectrometry and then searched in a database. We found the expression and upregulation of the toll-like receptor 4, ephx1 protein, isocitrate dehydrogenase, L-lactate dehydrogenase, GTP-binding nuclear protein Ran, and glyceraldehyde-3-phosphate dehydrogenase; however, the expression of the keratin type II cytoskeletal 8, type I cytokeratin, ARP3 actin-related protein 3 homolog, thyroid hormone receptor alpha-A, ATP synthase subunit beta, citrate synthase, tropomyosin 2, and tropomyosin 3 were downregulated. Six proteins were found in the hypoxia-inducible factor-1 (HIF-1) signaling pathway. We concluded that the grass carp gill is involved in response processes, including energy generation, metabolic processes, cellular structure, antioxidation, immunity, and signal transduction, to hypoxic stress. To our knowledge, this is the first study to conduct a proteomics analysis of expressed proteins in the gills of grass carp, and this study will help increase the understanding of the molecular mechanisms involved in hypoxic stress responses in fish at the protein level.

Background/aim: Nobiletin, a major polymethoxyflavones (PMFs) from citri reticulatae pericarpium (CRP), can inhibit several forms of cancer proliferation. However, the effects of nobiletin on nasopharyngeal carcinoma (NPC) C666-1 cells remain largely unknown., Materials and Methods: Cell counting kit 8 (CCK8) assay was used to measure cell vitality. Flow cytometry was performed to measure the apoptosis rate. Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot analysis were applied to determine the expression of mRNA and protein, respectively., Results: We showed that the proliferation rate of C666-1 cells was inhibited and the apoptosis rate was raised after treating with nobiletin. Moreover, nobiletin inhibited the expression of poly(ADP-ribose)polymerase-2 (PARP-2), and the tumor suppression effect of nobiletin on C666-1 is associated with PARP-2-dependent pathway., Conclusion: We demonstrated for the first time that nobiletin inhibited the growth of C666-1 cells, which may be relative to its regulation on PARP-2/SIRT1/AMPK signaling pathway. Our result implied that nobiletin may serve as a strategy to treat nasopharyngeal carcinoma., Competing Interests: The authors declare no conflict of interest.

A pot experiment was conducted to assess the phytoextraction potential for cadmium (Cd) of three types of sunflowers (edible, ornamental, and oil sunflowers) and the effects of the application of N, NP, and NPK fertilizers on Cd uptake of the three plants using Cd-contaminated soils collected from northern China. The results showed that fertilization could significantly increase the biomass, aboveground Cd concentrations and accumulation of ornamental and oil sunflowers, and the effect of NPK fertilizer was significantly better than those of other treatments. Cd concentrations of the aboveground parts of edible, ornamental, and oil sunflowers were 6.89, 8.92, 6.97 mg·kg -1 , respectively. Fertilization treatment significantly improved the transport ability of Cd of the three types of sunflowers, and bioconcentration factors of edible, ornamental, and oil sunflowers were 2.63 (control) to 3.10 (NPK fertilizer), 2.80 (control) to 4.02 (NPK fertilizer), and 2.11 (control) to 3.14 (NPK fertilizer), respectively. The results further showed that the metal-enriched granules and cellular debris were the main enrichment sources in the subcellular fraction of the three types of sunflowers (more than 55%). In summary, sunflowers can be considered as plant material for remediation of Cd-contaminated soil. In addition, NPK fertilizers can effectively improve the efficiency of sunflowers.

Air in concentrated animal feeding operations contains antibiotic resistance genes and airborne pathogens, with potential threat to human and animal health. In this study, air was sampled in the living area, outside, and inside of a fattening pig house in a pig farm for 24 and 48 hours. Feedstuffs, drinking water additives, and feces in the pig house were collected. Three kinds of antibiotic resistance genes (macrolide, β-lactam, and tetracycline) and seven pathogenic microorganisms (Campylobacter, Clostridium perfringens, Enterococcus, Escherichia coli, Yersinia enterocolitica, Staphylococcus spp., and Streptococcus suis) were detected by polymerase chain reaction (PCR). Six genes with high detection rates were selected, with their concentrations being determined by real-time quantitative PCR (qPCR). Results showed that three macrolide and two tetracycline resistance genes were detected in all air samples. Enterococcus, Escherichia coli, Yersinia enterocolitica, and Staphylococcus spp. were detected in air samples and drinking water additive. The concentrations of most target genes were above 10 4 copies·m -3 . The gene concentrations near the pig house were much higher than those in the living area. Main sources of antibiotic resistance genes and pathogens in the air were pig manure and drinking water additive. Sampling time of 24 h in the pig farm met the requirements for PCR detection. Sampling time of 48 h had a higher sampling efficiency than that of 24 h in the living area of the pig farm, whereas sampling time of 24 h was more appropriate than that of 48 h in high bioaerosol concentration area such as the pig house.

To enable the development of microbial agents and identify suitable candidate used for biodrying, the existence and function of Bacillus thermoamylovorans during sewage sludge biodrying merits investigation. This study isolated a strain of B. thermoamylovorans during sludge biodrying, submitted it for complete genome sequencing and analyzed its potential microbial functions. After biodrying, the moisture content of the biodrying material decreased from 66.33% to 50.18%, and B. thermoamylovorans was the ecologically dominant Bacillus, with the primary annotations associated with amino acid transport and metabolism (9.53%) and carbohydrate transport and metabolism (8.14%). It contains 96 carbohydrate-active- enzyme-encoding gene counts, mainly distributed in glycoside hydrolases (33.3%) and glycosyl transferases (27.1%). The virulence factors are mainly associated with biosynthesis of capsule and polysaccharide capsule. This work indicates that among the biodrying microorganisms, B. thermoamylovorans has good potential for degrading recalcitrant and readily degradable components, thus being a potential microbial agent used to improve biodrying., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Sewage sludge biodrying is a waste treatment method that uses bio-heat generated from organic degradation to remove moisture from sewage sludge. Lignocellulose and carbohydrate decomposition is important when assessing biodrying performance. This study investigated lignocellulose and carbohydrate decomposition, and the potential microbial functions during biodrying. We determined the lignocellulose and carbohydrate contents, assayed related enzyme activity, performed a complete metagenomic study on sewage sludge biodrying material during the thermophilic phase, annotated potential genetic function involved in the decomposition, and summarized the key metabolic pathways. The results indicated that lignocellulose, readily degradable carbohydrates, and starch, significantly decomposed after biodrying. During the thermophilic phase, the majority of lignocellulose and carbohydrate-related enzymes showed significantly higher activity, and glycoside hydrolases and glycosyl transferases showed higher gene counts and reads. Moreover, the top five microorganisms enriched with carbohydrate-active enzyme genes, i.e., Bacillus, Intrasporangium, Tetrasphaera, Rhodobacter, and Streptomyces, were also among the top ten ecologically dominant genera. These findings highlight the crucial phases for biodrying process, reveal the ecologically functional diversity of biodrying-originated microbial consortia, and suggest potential candidates for optimizing biodrying decomposition., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Midkine (Mdk) is a heparin-binding growth factor that is involved in regulating cell growth, differentiation and migration. Here, we report the isolation and characterization of duplicated mdk genes in blunt snout bream (Megalobrama amblycephala). The mdka and -b genes encode 146 aa and 147 aa peptides, respectively, sharing a sequence identity of 64%. During embryogenesis, mdka mRNA is detectable after 12 h post-fertilization (hpf) and mdkb mRNA can be detected after 8 hpf, about 4 h prior to mdka mRNA. Whole-mount in situ hybridization demonstrated that two paralogs of mdk mRNA were detected in the brain and dorsal neural tube at 16 hpf. At 22 hpf, mdka mRNA was abundant in the brain and dorsal neural tube, whereas mdkb mRNA were transcribed in the brain and tailbud. Later, at 55 hpf, both paralogs were mainly expressed in the brain. Furthermore, both the mdk genes were highly expressed in multiple adult tissues except in the skin and a low expression of mdka in the muscle. In addition, they were differentially inhibited in the liver and intestine with exogenous recombinant human growth hormone, while their mRNA levels were up-regulated in the brain. During starvation, both the mdk genes were significantly up-regulated in the intestine, brain and liver and returned to the control levels following 6 days of refeeding. Our results suggest that duplicated mdk genes may play conserved and divergent roles in embryonic development and tissue growth regulation in blunt snout bream., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Background: Inhibition of pancreatic lipase is an attractive approach to the treatment of obesity and other metabolic disorders. Naturally occurring phytochemicals are promising sources of lipase inhibitors., Purpose: In the present study, the anti-lipase activity of Citri Reticulatae Pericarpium (CRP) extracts was firstly evaluated in vitro. Moreover, the dynamic alteration of bioactive flavonoids in CRP collected at different time and its correlation with anti-lipase activities was investigated., Study Design/methods: Quantitative analysis of multi-components by a single-marker (QAMS) method was developed and validated for simultaneous determination of six flavonoids including narirutin, hesperidin, didymin, nobiletin, 3,5,6,7,8,3',4'-heptamethoxyflavone and tangeretin. Anti-lipase activity evaluation and docking studies of the flavonoids was also carried out to screen out the candidate lipase inhibitors., Results: The QAMS method validation results exhibited that the developed method had desirable specificity, linearity, precision and accuracy. CRP collected in early months contained higher concentrations of bioactive flavonoids, and exhibited more potent anti-lipase activity., Conclusion: Harvest timing had a significant impact on the amounts of bioactive flavonoids and the anti-lipase activities of CRP extracts. The contents of total flavonoids were positively correlated with the anti-lipase activities of CRP, and polymethoxyflavones played a significant role in the hypolipidemic effect of CRP. Nobiletin might be the most potential lipase inhibitor in CRP., (Copyright © 2018 Elsevier GmbH. All rights reserved.)

In Beijing, the chemical composition and component concentrations of volatile organic compounds (VOCs) were investigated during the municipal solid waste composting process using a portable gas chromatograph coupled with a mass spectrometer. The contributions of VOCs to the ozone formation potential were computed using the maximum incremental reactivity (MIR) scale and the propylene-equivalent concentration scale. The results showed that the concentrations of waste discharge in the sorting room, the first fermentation workshop, the second fermentation workshop, the compost product workshop, and the plant boundary were 10302.1, 15484.1, 929.9, 4693.6 and 370.4 μg·m -3 , respectively. The main VOCs of the municipal solid waste composting plant were ethanol, limonene, and acetone. The propylene-equivalent concentrations of waste discharge in the sorting room, the first fermentation workshop, the second fermentation workshop, the compost product workshop, and the plant boundary were 25875.7, 4087.9, 378.0, 747.7 and 296.8 μg·m -3 , whereas the O 3 formation potentials computed using the MIR scale were 26979.3, 21168.3, 1469.3, 6439.6 and 455.8 μg·m -3 . Reducing pollution by controlling the VOCs emission of waste discharge in the sorting room and the first fermentation workshop is important and can decrease the ozone formation potential. Given the accuracy and accessibility of the method, the MIR scale is more suitable for calculating the ozone formation potential of VOCs emitted from the municipal solid waste composting plant.

Purpose: This study explored the performance characteristics of a cuff-leak test (CLT) combined with interventional fiberoptic bronchoscopy (FBS) for evaluating whether early nasoendotracheal extubation was possible for patients who had received transoral atlantoaxial reduction plate (TARP) internal fixation surgery., Methods: 318 patients who underwent surgery were retrospectively analyzed (between January 2006 and December 2012). Extubation was performed by conventional approach (CA group, until December 2008) and improved approach (IA group, from January 2009) including CLT and an interventional FBS procedure. The extubation success within 1-3 days after surgery, incidence of postextubation stridor and tracheal reintubation were examined., Results: More IA-treated patients experienced extubation during the first 2 days than those CA-treated, median extubation time was 3 (2, 3) days in the CA group and 2 (1, 2) days in the IA group (all P < 0.01). The incidence of stridor and reintubation was 5.69 and 0.57 % in IA and 11.98 and 4.93 % in CA, respectively (both P < 0.05). For the CLT-positive patients in the IA group that remained intubated until day 3-4, interventional FBS was applied for safe extubation and achieved 100 % success., Conclusion: Early extubation through IA is safe and interventional FBS assists successful extubation for CLT-positive patients who underwent TARP surgery.

Four new polysaccharides (GPP-20, GPP-40, GPP-60 and GPP-80) were fractionated from Gynura procumbens leaves by 20%, 40%, 60% and 80% (v/v) ethanol, successively. Their physicochemical properties including the contents of neutral sugar, uronic acid and protein, as well as the monosaccharide composition were determined. In addition, the antioxidant activities of them were investigated via the reducing power assay and scavenging capacities of 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radicals and hydroxyl free radicals, respectively. The results indicated that apart from neutral sugar, they all contained uronic acids and proteins in their structures, which were further proved by the UV-vis and FT-IR spectra. Monosaccharide composition analysis implied that they all belonged to heteropolysaccharides consisted of arabinose, galactose, glucose, xylose and galacturonic acid with different types and ratios. What's more, GPP-20, GPP-40 and GPP-80 always exhibited better antioxidant activities than GPP-60 among these three antioxidant assays in vitro., (Copyright © 2016 Elsevier B.V. All rights reserved.)