RNA-seq analysis of small intestine transcriptional changes induced by starvation stress in piglets.

Piglets may experience a variety of stress injuries, but the molecular regulatory mechanisms underlying these injuries are not well understood. In this study, we analysed the ileum of Large White (LW) and Mashen (MS) piglets at different times of starvation using chemical staining and transcriptome analysis. The intestinal barrier of piglets was damaged after starvation stress, but the intestinal antistress ability of MS piglets was stronger than LW piglets. A total of 8021 differentially expressed genes (DEGs) were identified in two breeds. Interestingly, the immune capacity (CHUK, TLR3) of MS piglets increased significantly after short-term starvation stress, while energy metabolism (NAGS, PLA2G12B, AGCG8) was predominant in LW piglets. After long-term starvation stress, the level of energy metabolism (PLIN5, PLA2G12B) was significantly increased in MS piglets. The expression of immune (HLA-DQB1, IGHG4, COL3A1, CD28, LAT) and disease (HSPA1B, MINPPI, ADH1C, GAL3ST1) related genes were significantly increased in two breeds of piglets. These results suggest that short-term stress mainly enhances immunity and energy metabolism in piglets, while long-term starvation produces greater stress on piglets, making it difficult for them to compensate for the damage to their bodies through self-regulation. This information can help improve the stress resistance of piglets through molecular breeding.

Network Analysis of Depression Using Magnetoencephalogram Based on Polynomial Kernel Granger Causality.

Depression is a psychiatric disorder characterized by anxiety, pessimism, and suicidal tendencies, which has serious impact on human's life. In this paper, we use Granger causality index based on polynomial kernel as network node connectivity coefficient to construct brain networks from the magnetoencephalogram (MEG) of 5 depressed patients and 11 healthy individuals under positive, neutral, and negative emotional stimuli, respectively. We found that depressed patients had more information exchange between the frontal and occipital regions compared to healthy individuals and less causal connections in the parietal and central regions. We further analyzed the topological properties of the network revealed and found that depressed patients had higher average degrees under negative stimuli (p = 0.008) and lower average clustering coefficients than healthy individuals (p = 0.034). When comparing the average degree and average clustering coefficient of the same sample under different emotional stimuli, we found that depressed patients had a higher average degree and average clustering coefficient under negative stimuli than neutral and positive stimuli. We also found that the characteristic path lengths of patients under negative and neutral stimuli significantly deviated from small-world network. Our results suggest that the analysis of polynomial kernel Granger causality brain networks can effectively characterize the pathology of depression.

Molecular Mechanism of miR160d in Regulating Kiwifruit Resistance to Botrytis cinerea.

Gray mold caused by Botrytis cinerea leads to huge economic losses to the kiwifruit (Actinidia chinensis) industry. Elucidating the molecular mechanism responding to B. cinerea is the theoretical basis for the resistance to molecular breeding of kiwifruit. Previous studies have shown that miR160 regulates plant disease resistance through the indole-3-acetic acid (IAA) signaling pathway. In this study, kiwifruit "Hongyang" was used as the material, and Ac-miR160d and its target genes were identified and cloned. Overexpression and virus-induced gene silencing (VIGS) technology combined with RNA-seq were adopted to analyze the regulatory role of Ac-miR160d in kiwifruit resistance to B. cinerea. Silencing Ac-miR160d (AcMIR160d-KN) increased kiwifruit sensitivity to B. cinerea, whereas overexpression of Ac-miR160d (AcMIR160d-OE) increased kiwifruit resistance to B. cinerea, suggesting that Ac-miR160d positively regulates kiwifruit resistance to B. cinerea. In addition, overexpression of Ac-miR160d in kiwifruit increased antioxidant enzyme activities, such as catalase (CAT) and superoxide dismutase (SOD), and endogenous phytohormone IAA and salicylic acid (SA) content, in response to B. cinerea-induced stress. RNA-seq identified 480 and 858 unique differentially expressed genes in the AcMIR160d-KN vs CK and AcMIR160d-OE vs CK groups, respectively, with fold change ≥2 and false discovery rate <0.01. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed that families associated with "biosynthesis of secondary metabolites" are possibly regulated by Ac-miR160d. "Phenylpropanoid biosynthesis", "flavonoid biosynthesis", and "terpenoid backbone biosynthesis" were further activated in the two comparison groups upon B. cinerea infection. Our results may reveal the molecular mechanism by which miR160d regulates kiwifruit resistance to B. cinerea and may provide gene resources for molecular breeding in kiwifruit resistance.

Diversity and potential function of pig gut DNA viruses.

Viruses are ubiquitous in the gut of animals and play an important role in the ecology of the gut microbiome. The potential effects of these substances on the growth and development of the body are not fully known. Little is known about the effects of breeding environment on pig gut virome. Here, there are 3584 viral operational taxonomic units (vOTUs) longer than 5 kb identified by virus-enriched metagenome sequencing from 25 pig fecal samples. Only a small minority of vOTUs (11.16%) can be classified at the family level, and ∼50% of the genes could be annotated, supporting the concept of pig gut as reservoirs of substantial undescribed viral genetic diversity. The composition of pig gut virome in the six regions may be related to geography. There are only 20 viral clusters (VCs) shared among pig gut virome in six regions of Shanxi Province. These viruses rarely carry antibiotic resistance genes (ARGs). At the same time, they possess abundant auxiliary metabolic genes (AMGs) potentially involved in carbon, sulfur metabolism and cofactor biosynthesis, etc. This study has revealed the unique characteristics and potential function of pig gut DNA virome and established a foundation for the recognition of the viral roles in gut environment.

Effect of Reticulate Unit Spacing on Microstructure and Properties of Biomimetic 7075 Aluminum Alloy by Laser Cladding.

In the context of energy conservation and emission reduction, more and more attention has been paid to the development of lightweight metal materials with both high strength and high toughness. Inspired by the non-smooth surface of natural organisms, a biomimetic surface with various spacing reticulate units of 7075 aluminum alloys was modified by laser cladding. The microstructure, microhardness and tensile properties of the various spacing units with CeO2-SiC-Ni60 were studied. The finer microstructure and the higher microhardness of various spacing units in comparison with that of 7075 aluminum alloys were obtained, no matter the strip-like treated region or the cross-junction region. Moreover, the best combination of strength and toughness of the biomimetic sample with 2.5 mm spacing reticulate unit was discussed. Finally, by combining the microstructure, XRD phase change, thermal gradient effect, thermal expansion coefficient difference and hard phase strengthening mechanism, it was concluded that the 2.5 mm spacing reticulate unit had the best ability to inhibit crack propagation, and the dispersive hard phases of Al3Ni2 and SiC played a major role in stress release of the matrix.

Metagenomic analysis reveals patterns and hosts of antibiotic resistance in different pig farms.

In actual production environments, antibiotic-resistant genes (ARGs) are abundant in pig manure, which can form transmission chains through animals, the environment, and humans, thereby threatening human health. Therefore, based on metagenomic analysis methods, ARGs and mobile genetic elements (MGEs) were annotated in pig manure samples from 6 pig farms in 3 regions of Shanxi Province, and the potential hosts of ARGs were analyzed. The results showed that a total of 14 ARG types were detected, including 182 ARG subtypes, among which tetracycline, phenol, aminoglycoside, and macrolide resistance genes were the main ones. ARG profiles, MGE composition, and microbial communities were significantly different in different regions as well as between different pig farms. In addition, Anaerobutyricum, Butyrivibrio, and Turicibacter were significantly associated with multiple ARGs, and bacteria such as Prevotella, Bacteroides, and the family Oscillospiraceae carried multiple ARGs, suggesting that these bacteria are potential ARG hosts in pig manure. Procrustes analysis showed that bacterial communities and MGEs were significantly correlated with ARG profiles. Variation partitioning analysis results indicated that the combined effect of MGEs and bacterial communities accounted for 64.08% of resistance variation and played an important role in ARG profiles. These findings contribute to our understanding of the dissemination and persistence of ARGs in actual production settings, and offer some guidance for the prevention and control of ARGs contamination.

Integrated Omics Analysis Reveals Alterations in the Intestinal Microbiota and Metabolites of Piglets After Starvation.

Obesity is a serious public health problem. Short-term starvation is an effective way to lose weight but can also cause harm to the body. However, a systematic assessment of the relationship between the intestinal microbiota and metabolites after complete fasting is lacking. Pigs are the best animal models for exploring the mechanisms of human nutrition digestion and absorption, metabolism, and disease treatment. In this study, 16S rRNA sequencing and liquid chromatography-mass spectrometry were used to analyze the changes in the intestinal microbiota and metabolite profiles in piglets under starvation stress. The results show that the microbial composition was changed significantly in the starvation groups compared with the control group (P < 0.05), suggesting that shifts in the microbial composition were induced by starvation stress. Furthermore, differences in the correlation of the intestinal microbiota and metabolites were observed in the different experimental groups. Starvation may disrupt the homeostasis of the intestinal microbiota and metabolite profile and affect the health of piglets. However, piglets can regulate metabolite production to compensate for the effects of short-term starvation. Our results provide a background to explore the mechanism of diet and short-term hunger for intestinal homeostasis.

Effects of Maternal Factors and Postpartum Environment on Early Colonization of Intestinal Microbiota in Piglets.

Intestinal microbiota significantly influences the intake, storage, and utilization of body nutrients, as well as animal growth and development. The establishment of microbiota is affected by many factors, such as delivery and feeding modes, antibiotics, disease, and the surrounding environment. In this study, we selected Chinese indigenous Mashen and Jinfen White pigs as the study subjects. To explore the source and factors affecting the piglet intestinal microbiota, 16S rRNA gene sequencing was performed to analyze the microbial composition of the feces, saliva, vaginal secretions, and colostrum of parturient sows, feces and saliva of newborn piglets, and surrounding environment samples. The results showed that the microbiota of the saliva of sows and piglets is structurally similar to that of the environment and is dominated by the phylum Proteobacteria, including Acinetobacter, Actinomyces, and Pseudomonas. The core genus in the vaginal secretions and colostrum of sows was Pseudomonas. Among the fecal samples, the core bacterial genera in sows before and after delivery were Clostridium sensu_stricto_1 and Christensenellaceae_R-7_group, while in piglets at 1 d of age, Pseudomonas and Escherichia-Shigella were most abundant. These results indicate that microbiota in feces, colostrum, and vaginal secretions of sows more easily colonized piglet intestines through a symbiotic effect. The environmental and salivary microbiota could also affect the early colonization and succession of the intestinal microbiota of piglets to some extent. This study provides a theoretical basis for sow delivery protection and early nursing of piglets and background for the research and development of microbial agents to improve piglet intestinal health.

Psoralen Suppresses Lipid Deposition by Alleviating Insulin Resistance and Promoting Autophagy in Oleate-Induced L02 Cells.

Non-alcoholic fatty liver disease (NAFLD) held a high global prevalence in recent decades. Hepatic lipid deposition is the major characteristic of NAFLD. We aim to explore the mechanisms of psoralen on lipid deposition in NAFLD. The effects of psoralen on insulin resistance, lipid deposition, the expression and membrane translocation of glucose transporter type 4 (GLUT4), autophagy, and lipogenesis enzymes were determined on sodium oleate-induced L02 cells. Chloroquine and 3-MA were employed. The AMP-activated protein kinase alpha (AMPKα) was knocked down by siRNA. Psoralen alleviated insulin resistance in sodium oleate-induced L02 hepatocytes by upregulating the expression and membrane translocation of GLUT4. Psoralen inhibited lipid accumulation by decreasing the expression of key lipogenesis enzymes. Psoralen promotes autophagy and the autophagic flux to enhance lipolysis. Psoralen promoted the fusion of the autophagosome with the lysosome. Both chloroquine and 3-MA blocked the effects of psoralen on autophagy and lipid accumulation. The AMPKα deficiency attenuated the effects of psoralen on autophagy and lipid accumulation. Our study demonstrated that as an antioxidant, psoralen attenuates NAFLD by alleviating insulin resistance and promoting autophagy via AMPK, suggesting psoralen to be a promising candidate for NAFLD.

Icaritin alleviates docetaxel-induced skin injury by suppressing reactive oxygen species via estrogen receptors.

BACKGROUND: Docetaxel (DTX) exhibits antitumor effects against breast cancer by stabilizing microtubules and increasing the accumulation of reactive oxygen species (ROS). DTX extravasation during infusion often causes skin injury. The present study aimed to investigate the effects and mechanisms of icaritin (ICT) on DTX-induced skin injury. METHODS: The effects of ICT on the viability and apoptosis of HaCaT cells were measured by SRB assay and flow cytometry, respectively. Endogenous LC3 puncta and microtubules were determined by immunofluorescence. The number of mitochondria was measured by MitoTracker orange staining. ROS were determined by dihydroethidium staining. The expression of markers of ROS and autophagy were measured by western blotting. Chloroquine, compound D, and tamoxifen were employed as the inhibitor for autophagy and AMPK, estrogen receptors (ERs) modulator, respectively. RESULTS: DTX inhibited the viability and decreased apoptosis of HaCaT cells, which can be rescued by ICT. ICT decreased microtubule bundles, increased the number of mitochondria, and attenuated ROS of HaCaT cells induced by DTX. ICT blocks autophagy and the autophagic flux. Compound C or tamoxifen diminished the protection effects of ICT on DTX-treated HaCaT cells. CONCLUSION: ICT alleviates DTX-induced skin injury by suppressing ROS, reducing microtubule bundles, and blocking autophagy via ERs. Our study indicated that ICT may be a potential candidate for DTX-induced skin injury.

Comparative Evaluation of the Ileum Microbiota Composition in Piglets at Different Growth Stages.

Intestinal microbiota can affect the intake, storage, and absorption of nutrients in the body, thereby greatly impacting the growth and development of animals. In addition to diet, the breed and growth stages of pigs could also affect changes in the intestinal microbiota. However, research on the developmental changes in the ileum microbiota of piglets remains unclear. In this study, the ileum microbiota of Jinfen White and Mashen piglets at different developmental stages were investigated using 16S rRNA sequencing. Physiologically, the villus height of the ileum decreased, and the crypt depth increased during the development of the two pig breeds. Additionally, the serum antioxidant factors in the Jinfen White piglets were significantly higher than in the Mashen piglets at the end of the nursing stage. A total of 690 operational taxonomic units (OTUs) belonging to 21 phyla and 286 genera were identified, of which Firmicutes and Proteobacteria were the dominant phyla during the development of both the Jinfen White and Mashen piglets, accounting for ∼90% of all OTUs. Further research revealed differences in dominant bacteria between the two breeds. With increasing age, the ileum microbial diversity increased, and in both the pig breeds, the proportion of Firmicutes increased, whereas the proportion of Proteobacteria decreased. Additionally, different samples were characterized by specific genera, and different Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways were predicted at certain developmental stages. Finally, the correlation between the ileum microbiota and physiological features was analyzed, and it was suggested that the host and environmental factors play important roles in the formation of the microbial community structure in piglets. In summary, we delineated the structure, function, and differences in ileum microbiota between Jinfen White and Mashen piglets during different growth stages. This study helps to understand the development of the intestinal microbiota in local and hybrid pig breeds.

A double-edged sword: The Kelch-like ECH-associated protein 1-nuclear factor erythroid-derived 2-related factor 2-antioxidant response element pathway targeted pharmacological modulation in nonalcoholic fatty liver disease.

Nutraceuticals activating the Kelch-like epichlorohydrin (ECH)-associated protein 1 (Keap1)-nuclear factor erythroid-derived 2-related factor 2 (Nrf2)-antioxidant response element (ARE) pathway are widely used for nonalcoholic fatty liver disease (NAFLD) because no specific drugs are approved yet. The pathology of NAFLD is summarized as the 'two-hit' hypothesis. The 'first hit' includes insulin resistance and lipid accumulation. Oxidative stress, lipid peroxidation, and inflammation are regarded as the 'second hit'. Now there is controversial evidence about the roles of the Keap1-Nrf2-ARE pathway and its activators in NAFLD. When the 'first hit' occurs, the hepatocyte-specific Nrf2 deficiency reduces insulin resistance and significantly attenuates lipid accumulation. However, when the 'second hit' occurs, Nrf2 activation reduces oxidative stress and combats inflammation. We reviewed the roles of the Keap1-Nrf2-ARE pathway as a double-edged sword in the development of NAFLD, its inhibitors as a novel therapeutic approach for early NAFLD, and the nutraceutical character of its activators.

Mechanism and Catalysis of Oxidative Degradation of Fiber-Reinforced Epoxy Composites.

Carbon fiber-reinforced polymer (CFRP) materials are widely used in aerospace and recreational equipment, but there is no efficient procedure for their end-of-life recycling. Ongoing work in the chemistry and engineering communities emphasizes recovering carbon fibers from such waste streams by dissolving or destroying the polymer binding. By contrast, our goal is to depolymerize amine-cured epoxy CFRP composites catalytically, thus enabling not only isolation of high-value carbon fibers, but simultaneously opening an approach to recovery of small molecule monomers that can be used to regenerate precursors to new composite resin. To do so will require understanding of the molecular mechanism(s) of such degradation sequences. Prior work has shown the utility of hydrogen peroxide as a reagent to affect epoxy matrix decomposition [1]. Herein we describe the chemical transformations involved in that sequence: the reaction proceeds by oxygen atom transfer to the polymer's linking aniline group, forming an N-oxide intermediate. The polymer is then cleaved by an elimination and hydrolysis sequence. We find that elimination is the slower step. Scandium trichloride is an efficient catalyst for this step, reducing reaction time in homogeneous model systems and neat cured matrix blocks. The conditions can be applied to composed composite materials, from which pristine carbon fibers can be recovered.

Sulforaphane Prevents Neuronal Apoptosis and Memory Impairment in Diabetic Rats.

BACKGROUND/AIMS: To explore the effects of sulforaphane (SFN) on neuronal apoptosis in hippocampus and memory impairment in diabetic rats. METHODS: Thirty male rats were randomly divided into normal control, diabetic model and SFN treatment groups (N = 10 in each group). Streptozotocin (STZ) was applied to establish diabetic model. Water Morris maze task was applied to test learning and memory. Tunel assaying was used to detect apoptosis in hippocampus. The expressions of Caspase-3 and myeloid cell leukemia 1(MCL-1) were detected by western blotting. Neurotrophic factor levels and AKT/GSK3ß pathway were also detected. RESULTS: Compared with normal control, learning and memory were apparently impaired, with up-regulation of Caspase-3 and down-regulation of MCL-1 in diabetic rats. Apoptotic neurons were also found in CA1 region after diabetic modeling. By contrast, SFN treatment prevented the memory impairment, decreased the apoptosis of hippocampal neurons. SFN also attenuated the abnormal expression of Caspase-3 and MCL-1 in diabetic model. Mechanically, SFN treatment reversed diabetic modeling-induced decrease of p-Akt, p-GSK3ß, NGF and BDNF expressions. CONCLUSION: SFN could prevent the memory impairment and apoptosis of hippocampal neurons in diabetic rat. The possible mechanism was related to the regulation of neurotropic factors and Akt/GSK3ß pathway.