Exposure to high concentrations of triphenyl phosphate altered functional performance, liver metabolism and intestinal bacterial composition of aquatic turtles.

Organophosphorus flame retardants, such as triphenyl phosphate (TPhP), exist ubiquitously in various environments owing to their widespread usage. Potential toxic effects of residual flame retardants on cultured non-fish species are not concerned commonly. TPhP-induced physiological and biochemical effects in an aquatic turtle were evaluated here by systematically investigating the changes in growth and locomotor performance, hepatic antioxidant ability and metabolite, and intestinal microbiota composition of turtle hatchlings after exposure to different TPhP concentrations. Reduced locomotor ability and antioxidant activity were only observed in the highest concentration group. Several metabolic perturbations that involved in amino acid, energy and nucleotide metabolism, in exposed turtles were revealed by metabolite profiles. No significant among-group difference in intestinal bacterial diversity was observed, but the composition was changed markedly in exposed turtles. Increased relative abundances of some bacterial genera (e.g., Staphylococcus, Vogesella and Lawsonella) probably indicated adverse outcomes of TPhP exposure. Despite having only limited impacts of exposure at environmentally relevant levels, our results revealed potential ecotoxicological risks of residual TPhP for aquatic turtles considering TPhP-induced metabolic perturbations and intestinal bacterial changes.

Environmentally Relevant Levels of Antiepileptic Carbamazepine Altered Intestinal Microbial Composition and Metabolites in Amphibian Larvae.

There is growing concern about the potential ecological risks posed by pharmaceutical residues in the aquatic environment. However, our understanding of the toxic effects of antiepileptic pharmaceuticals, such as carbamazepine (CBZ), on aquatic animal larvae is still limited. In this study, the tadpoles of the black-spotted pond frog (Pelophylax nigromaculatus) were exposed to environmentally relevant concentrations of CBZ (0.3 and 3.0 µg/L) for 30 days, and their growth, intestinal microbial composition, and metabolites were investigated to assess the potential toxic effects of CBZ in non-targeted aquatic organisms. Some tadpoles died during exposure, but there was no significant among-group difference in the survival and growth rates. CBZ exposure significantly altered the composition of tadpole intestinal microbiota. Relative abundances of some bacterial genera (e.g., Blautia, Prevotella, Bacillus, Microbacterium, etc.) decreased, while others (e.g., Paucibacter, etc.) increased in CBZ-exposed tadpoles. Interestingly, CBZ-induced alterations in some bacteria might not necessarily lead to adverse outcomes for animals. Meanwhile, small molecular intestinal metabolites related to energy metabolism, and antioxidant and anti-inflammatory activities were also altered after exposure. Taken together, environmentally relevant levels of CBZ might alter the metabolic and immune performances of amphibian larvae by modifying the abundance of some specific bacteria and the level of metabolites in their intestines, thereby potentially causing a long-term effect on their fitness.

Effects of Habitual Dietary Change on the Gut Microbiota and Health of Silkworms.

Diet plays a crucial role in shaping the gut microbiota and overall health of animals. Traditionally, silkworms are fed fresh mulberry leaves, and artificial diets do not support good health. The aim of this study was to explore the relationship between the dietary transition from artificial diets to mulberry leaves and the effects on the gut microbiota and physiological changes in silkworms as a model organism. With the transition from artificial diets to mulberry leaves, the diversity of the silkworm gut microbiota increased, and the proportion of Enterococcus and Weissella, the dominant gut bacterial species in silkworms reared on artificial diets, decreased, whereas the abundance of Achromobacter and Rhodococcus increased. Dietary transition at different times, including the third or fifth instar larval stages, resulted in significant differences in the growth and development, immune resistance, and silk production capacity of silkworms. These changes might have been associated with the rapid adaptation of the intestinal microbiota of silkworms to dietary transition. This study preliminarily established a dietary transition-gut microbial model in silkworms based on the conversion from artificial diets to mulberry leaves, thus providing an important reference for future studies on the mechanisms through which habitual dietary changes affect host physiology through the gut microbiome.

Embryonic development, hatchling performance and metabolic profile after egg exposure to environmentally relevant levels of chlorpyrifos in an aquatic turtle.

The extensive use of organophosphorus insecticides poses a threat to the survival of non-target organisms. Ecotoxicological outcomes of embryonic exposure to insecticides are rarely evaluated in various oviparous species. In this study, soft-shelled turtle (Pelodiscus sinensis) eggs were incubated in moist substrate containing different levels (0, 2, 20 and 200 µg/kg) of chlorpyrifos to investigate its toxic effects on embryonic development and survival, and hatchling physiological performance. Chlorpyrifos exposure had no significant impacts on embryonic development rate and egg survival in P. sinensis. Similarly, embryonic chlorpyrifos exposure neither obviously affected the size and locomotor performance of hatchlings, nor changed the activities of superoxide dismutase and catalase, and content of malondialdehyde in their erythrocytes. Based on liquid chromatography-mass spectrometry analysis, minor metabolic perturbations related to amino acid, lipid and energy metabolism in hatchlings after embryonic chlorpyrifos exposure were revealed by hepatic metabolite profiling. Overall, our results suggested that embryonic exposure to environmentally relevant levels of chlorpyrifos had only a limited impact on physiological performances of hatchlings, although it would result in a potential risk of hepatotoxicity in P. sinensis.

miR-148a-3p regulates proliferation and apoptosis of bovine muscle cells by targeting KLF6.

Skeletal muscle development is regulated by a series of regulatory factors, and also including noncoding RNA, especially microRNAs (miRNAs). Recently, miR-148a has been found to be involved in murine C2C12 differentiation by targeting ROCK1. However, the function of miR-148a-3p for the proliferation and apoptosis of bovine muscle cells has not been determined. In this study, we found that miR-148a-3p was highly expressed in fetal bovine skeletal muscle and exhibited a decreasing trend in muscle cells during its growth phase. Functional studies indicated that gain of miR-148a-3p inhibited the proliferation of bovine muscle cells and promoted apoptosis. Conversely, interference with miR-148a-3p inhibitor promoted muscle cell proliferation and inhibited its apoptosis. Mechanistically, KLF6 was confirmed as a new potential target gene of miR-148a-3p by TargetScan software prediction and the dual-luciferase assay verification. Additionally, after a gain or loss of KLF6, the function of KLF6 for muscle cell proliferation and apoptosis was opposite to that of miR-148a-3p. Collectively, these findings proposed a novel avenue whereby miR-148a-3p impeded bovine myoblast cell proliferation and promoted apoptosis through the posttranscriptional downregulation of KLF6.

ΔFosB regulates rosiglitazone-induced milk fat synthesis and cell survival.

Rosiglitazone induces adipogenesis in adipocyte and regulates cell survival and differentiation in number of cell types. However, whether PPARγ regulates the synthesis of milk fat and cell survival in goat mammary gland remains unknown. Rosiglitazone strongly enhanced cellular triacylglycerol content and accumulation of lipid droplet in goat mammary epithelial cells (GMEC). Furthermore, ΔFosB decreased the expression of PPARγ at both mRNA and protein levels, and rosiglitazone-induced milk fat synthesis was abolished by ΔFosB overexpression. ΔFosB reduced milk fat synthesis and enhanced saturated fatty acid concentration. Rosiglitazone increased the number of GMEC in G0/G1 phase and inhibited cell proliferation, and these effects were improved by overexpression of ΔFosB. ΔFosB was found to promote the expression of Bcl-2 and suppress the expression of Bax, and protected GMEC from apoptosis induced by rosiglitazone. Intracellular calcium trafficking assay revealed that rosiglitazone markedly increased intracellular calcium concentration. ΔFosB protected GMEC from apoptosis induced by intracellular Ca2+ overload. ΔFosB increased MMP-9 gelatinolytic activity. SB-3CT, an MMP-9 inhibitor, suppressed the expression of Bcl-2, and increased intracellular calcium levels, and this effect was abolished by ΔFosB overexpression. SB-3CT induced GMEC apoptosis and this effect was inhibited by ΔFosB overexpression. These findings suggest that ΔFosB regulates rosiglitazone-induced milk fat synthesis and cell survival. Therefore, ΔFosB may be an important checkpoint to control milk fat synthesis and cell apoptosis.

CircFUT10 reduces proliferation and facilitates differentiation of myoblasts by sponging miR-133a.

Circular RNAs (circRNAs) have been identified in various tissues and cell types from human, monkey, porcine, and mouse. However, knowledge on circRNAs in bovine muscle development is limited. We downloaded and analyzed the circRNAs sequencing data of bovine skeletal muscle tissue, and further characterized the role of a candidate circRNA (circFUT10) in muscle development. Quantitative real-time PCR (qPCR) and Western blot assays were used to confirm the expression of genes involved in myoblasts differentiation and proliferation. Flow cytometry was performed to assess cell cycle distribution and cell apoptosis. EdU incorporation and CCK-8 assay were performed to demonstrate cell proliferation. We demonstrated that circFUT10 was highly (but differentially) expressed in embryonic and adult skeletal muscle tissue. circFUT10 induced bovine primary myoblasts differentiation and increased the expression of MyoD, MyoG, and MyhC in mRNA and protein levels. circFUT10 increased the number of myoblasts in the G0/G1 phase of the cell cycle, and decreased the proportion of cells in the S-phase. circFUT10 inhibited the proliferation of myoblasts and promoted them apoptosis. Via a luciferase screening assay, circFUT10 is observed to sponge to miR-133a with three potential binding sites. Specifically, we show that circFUT10 regulated myoblasts differentiation and cell survival by directly binding to miR-133a and inhibiting miR-133a activity. Modulation of circFUT10 expression in muscle tissue may emerge as a potential target in breeding strategies attempting to control muscle development in cattle.

Understanding the spatial and temporal patterns of copper in-use stocks in China.

Two approaches are adopted to characterize the comprehensive pattern of the copper in-use stocks in China. The top-down results indicate that both the total amount and the per capita quantity of the stocks have exhibited a significant and increasing trend for the past 60 years, especially since 2000. The top-down results show that the copper stocks increased from a negligible level of less than 1 kg/capita in 1952 to 44 kg/capita in 2012. The total stocks in 2012 are estimated to be 60 Mt by a top-down approach or 48 Mt by a bottom-up calculation. The bottom-up method determines that the largest reservoir is the infrastructure sector, which accounts for approximately 58% of the total stocks. The spatial pattern indicates that the copper in-use stocks are predominately spatially distributed in the eastern regions of China, a feature that is obviously different from the geographical distribution of the primary resources. Analysis on the prospects of stocks shows both the total magnitude and per capita value will continuously increase in the following decade, and enter a relatively stable stage in around 2030, with a maximum value of 106 kg/capita. The results improve the knowledge about closing copper cycles.

Hepatic Metabolomic Responses to Low-Temperature Stress in the Invasive Turtle, Trachemys scripta elegans.

Investigating the physiological and biochemical changes of ectothermic species before entering hibernation would contribute to the understanding of how they adapt to low-temperature environments. Here, red-eared slider turtle (Trachemys scripta elegans) hatchlings were maintained under different thermal treatments (24 °C, slowly decreasing temperatures from 24 °C to 14 °C, and to 4 °C). Hepatic metabolite alterations were measured to assess the metabolic impacts of low-temperature stress in this species. Of these differentially changed metabolites, some (e.g., raffinose, spermidine, allocholic acid, taurohyocholate, 2-ketobutyric acid, acetylcysteine) were shown to decrease, while others (e.g., stearolic acid, D-mannose) increased in low-temperature treatments. Our results indicated that short-term low-temperature stress might have limited impacts on lipid and energy metabolism in this species. The changes in other metabolites (e.g., allocholic acid, taurohyocholate, spermine, acetylcysteine) might be associated with a low food intake (and thus reduced digestive performance) and weakened immune ability of low-temperature-exposed animals.

Nonsteaming method improves the nutritional value and utilization efficiency of silkworm artificial diets.

Artificial diets for silkworms overcome the seasonal limitations of traditional rearing methods with fresh mulberry leaves. However, the current wet artificial diets, steamed at high temperatures, are not favored by silkworms, and they are cumbersome and challenging to preserve. These conditions adversely affected the development of artificial diet-based sericulture production. In this study, we disinfected dry powder diets with radiation and added distilled water without steaming before use. Then, the nutritional value of finished diets and their impact on silkworm development was assessed. Compared with steamed diets, nonsteamed diets were more attractive to silkworms. Chemical assays showed significantly more essential nutrients for silkworms, including l-ascorbic acid, vitamin B1, vitamin B2, and urease in nonsteamed diets than in steamed diets. Feeding fifth-instar silkworm larvae with nonsteamed diets significantly improved the ammonia utilization efficiency of the diet and increased the cocoon shell rate and diet/silk protein conversion efficiency by 5.9% and 13.3%, respectively. When fed with nonsteamed diets, the abundance of aerobic microorganisms in silkworm intestines increased and the abundance of pathogenic bacteria decreased. Furthermore, the vitality of the silkworm, measured by the dead worm cocoon rate, significantly improved by 16.90%. In summary, preparing sterile wet diets without high-temperature steaming effectively improved the nutritional value of the diet and enhanced silkworm growth.

Environmental Pollution Liability Insurance Pricing and the Solvency of Insurance Companies in China: Based on the Black-Scholes Model.

Environmental pollution liability insurance is becoming increasingly important for China to achieve its emission reduction targets. Insurance pricing is a crucial factor restricting the market share of environment pollution liability insurance, from the perspective of the Black-Scholes pricing model, which in turn has influenced the solvency of insurance companies in China. Firstly, this study analyzes the problems existing in compulsory liability insurance for environmental pollution in China. It proceeds with analyzing the price of compulsory environmental pollution liability insurance using the Black-Scholes pricing model, and derives a high premium insurance rate of 2.44%. Moreover, it performs a multivariate regression analysis using the asset and liability data, taken from the annual report, to identify three key factors affecting the solvency adequacy ratio, namely, capital debt ratio, reflecting the company asset structure; net interest rate on assets, reflecting the asset scale with actual solvency; and claim ratio, reflecting the business quality. Based on the results of regression analysis and robustness test for the China Insurance Clauses (CIC) company, People's Insurance Company of China (PICC), and Asia-Pacific Property & Casualty Insurance (API) company, it is shown that the effect of total asset, total debt, capital debt ratio, claim ratio, and net interest rate on assets on the solvency adequacy ratio is significant, with respect to the size of the coefficients. Based on the Black-Scholes pricing model found in the previous cycle of liability insurance, and keeping in view the existing problems of environmental pollution liability insurance expenditure, this paper presents suggestions that are conducive to improving the solvency of insurance companies in China.

Risk Factors for Recurrence After Percutaneous Endoscopic Lumbar Discectomy: A Meta-Analysis.

OBJECTIVE: Postoperative recurrence of percutaneous endoscopic lumbar disc increases the physical damage and financial burden on patients and negatively affects physicians' treatment decisions. We conducted this meta-analysis to explore the risk factors for postoperative recurrence of percutaneous endoscopic lumbar disc for lumbar disc herniation. METHODS: We conducted article search in the PubMed, EMBASE and Cochrane Library databases. PRISMA guidelines were followed in this review. The data are statistically analyzed by the Roundup Manager (version 3.6.1). The results of the meta-analysis are presented in the form of forest-like plots. RESULTS: We included 13 articles and identified 7524 cases. Patients with older age (odds ratio [OR] = 1.28, 95% confidence interval [CI]: 1.18-1.40), higher body mass index (OR = 1.16, 95% CI: 1.05-1.28), smoker (OR = 1.73, 95% CI: 0.95-3.15), degenerative grades ≥3 (OR = 6.07, 95% CI: 2.81-13.11), and postoperative sagittal motion ≥10° (OR = 2.42, 95% CI: 1.63-3.58) have a higher recurrence rate. CONCLUSIONS: A thorough preoperative evaluation is essential to prevent postoperative recurrence of percutaneous endoscopic lumbar discectomy. The study addresses several factors of preoperative evaluation, which is hopeful to provide a reference for neurosurgeons.

Exposure to low levels of antidiabetic glibenclamide had no evident adverse effects on intestinal microbial composition and metabolic profiles in amphibian larvae.

Unmetabolized human pharmaceuticals may enter aquatic environments, and potentially exert adverse effects on the survival of non-target organisms. Here, Pelophylax nigromaculatus tadpoles were exposed to different concentrations of antidiabetic glibenclamide (GLB) for 30 days to evaluate its potential ecotoxicological effect in amphibians using intestinal microbiomic and metabolomic profiles. The mortality rate of GLB-exposed groups appeared to be lower than that of the control group. Despite not being statistically significant, there was a tendency for a decrease in intestinal microbial diversity after exposure. The relative abundance of bacteria phylum Firmicutes was shown to decrease, but those of other phyla did not in GLB-exposed tadpoles. Some potentially pathogenic bacteria (e.g., Clostridium, Bilophila, Hafnia) decrease unexpectedly, while some beneficial bacteria (e.g., Akkermansia, Faecalibacterium) increased in GLB-exposed tadpoles. Accordingly, GLB-induced changes in intestinal microbial compositions did not seem harmful to animal health. Moreover, minor changes in a few intestinal metabolites were observed after GLB exposure. Overall, our results suggested that exposure to low levels of GLB did not necessarily exert an adverse impact on amphibian larvae.

Sericin Ser3 Ectopic Expressed in Posterior Silk Gland Affects Hemolymph Immune Melanization Response via Reducing Melanin Synthesis in Silkworm.

The transgenesis of silkworms is an important way to innovate genetic resources and silk function. However, the silk-gland (SG) of transgenic silkworms, which is the most concerned target tissue of sericulture, often suffers from low vitality, stunting and other problems, and the reasons are still unknown. This study trans engineered recombinant Ser3, a middle silk gland (MSG) specific expression gene, in the posterior silk gland (PSG) of the silkworm, and studied hemolymph immune melanization response changes in mutant pure line SER (Ser3+/+). The results showed that although the mutant had normal vitality, the melanin content and phenoloxidase (PO) activity in hemolymph related to humoral immunity were significantly reduced, and caused significantly slower blood melanization and weaker sterilization ability. The mechanism investigation showed that the mRNA levels and enzymatic activities of phenylalanine hydroxylase (PAH), tyrosine hydroxylase (TH) and dopamine decarboxylase (DDC) in the melanin synthesis pathway in mutant hemolymph, as well as the transcription levels of the PPAE, SP21 and serpins genes in the serine protease cascade were significantly affected. Moreover, the total antioxidant capacity, superoxide anion inhibition capacity and catalase (CAT) level related to the redox metabolic capacity of hemolymph were significantly increased, while the activities of superoxide dismutase (SOD) and glutathione reductase (GR), as well as the levels of hydrogen peroxide (H2O2) and glutathione (GSH), were significantly decreased. In conclusion, the anabolism of melanin in the hemolymph of PSG transgenic silkworm SER was inhibited, while the basic response level of oxidative stress was increased, and the hemolymph immune melanization response was decreased. The results will significantly improve the safe assessment and development of genetically modified organisms.

Determinants of Proactive Low-Carbon Consumption Behaviors: Insights from Urban Residents in Eastern China.

Proactive low-carbon consumption behaviors (PLCBs) are crucial to achieving carbon neutrality and identifying motivations for PLCBs is indispensable to changing individual consumption patterns. This study establishes a model by incorporating individual-group-level factors with psychological empowerment perception. The ordinary least-squares regression model was applied to identify the influencing factors of PLCBs with data collected from 1732 urban residents in eastern China. Results show that PLCBs are positively influenced by normative internalization, learning capacity, symbol concern, expertise level, and an environmentalism culture. In particular, the effect of learning capacity is the largest, with an influence coefficient of 0.271. A negative impact is observed between the consumerism culture and PLCBs. Moreover, psychological empowerment perception partly medicated the association between individual-group-level factors and PLCBs, and the maximum ratio of mediating effect to the full impact is 62.64%. The study sheds light on low-carbon-related behavioral management, and recommendations to promote PLCBs are further proposed.

miR-183/96/182 Cluster Regulates the Development of Bovine Myoblasts through Targeting FoxO1.

Muscle development is an important factor affecting meat yield and quality and is coordinated by a variety of the myogenic genes and signaling pathways. Recent studies reported that miRNA, a class of highly conserved small noncoding RNA, is actively involved in regulating muscle development, but many miRNAs still need to be further explored. Here, we identified that the miR-183/96/182 cluster exhibited higher expression in bovine embryonic muscle; meanwhile, it widely existed in other organizations. Functionally, the results of the RT-qPCR, EdU, CCK8 and immunofluorescence assays demonstrated that the miR-183/96/182 cluster promoted proliferation and differentiation of bovine myoblast. Next, we found that the miR-183/96/182 cluster targeted FoxO1 and restrained its expression. Meanwhile, the expression of FoxO1 had a negative correlation with the expression of the miR-183/96/182 cluster during myoblast differentiation. In a word, our findings indicated that the miR-183/96/182 cluster serves as a positive regulator in the proliferation and differentiation of bovine myoblasts through suppressing the expression of FoxO1.

Effects of Dietary Lysine Levels on Growth Performance, Nutrient Digestibility, Serum Metabolites, and Meat Quality of Baqing Pigs.

This study was carried out to determine the Lys requirements of Baqing pigs and the effects of different dietary lysine levels on growth performance, apparent nutrient digestibility, serum metabolites, and carcass and meat traits. A total of 120 castrated Baqing pigs were selected by body weight and randomly assigned to five dietary treatments with six replicate pens (4 pigs per pen, castrated) per treatment in a randomized complete block design. Five diets in mash form were formulated to contain SID Lys at 0.56%, 0.68%, 0.80%, 0.92%, and 1.04% of diet in phase 1 (20−40 kg), at 0.45%, 0.54%, 0.63%, 0.72%, and 0.81% of diet in phase 2 (40−60 kg), and at 0.39%, 0.47%, 0.55%, 0.63%, and 0.71% of diet in phase 3 (60−90 kg), respectively. The results showed that the bodyweight of pigs was not affected by dietary SID Lys content during each period. However, the addition of dietary SID Lys linearly reduced F/G in the first period and quadratically increased ADG during the second period (p < 0.05). The digestible energy (DE) was increased linearly and quadratically in the first phases with the dietary increased SID Lys levels, while DE was reduced in the third and second phases (p < 0.05). Increasing SID Lys contents linearly increased the serum TG concentration and quadratically decreased the serum GLU concentration, while linearly reducing the serum HDLC concentration of first period pigs (p < 0.05). Serum concentrations of TP, TG, TC, and LDLC were increased linearly with the increasing dietary SID Lys levels in the second period (p < 0.05). The serum concentrations of Lys increased quadratically, and histidine increased linearly with the increased dietary SID Lys levels (p < 0.05). Compared with the treatment three group, dietary SID Lys addition content at treatment four increased the shear force of the longissimus dorsi muscle (p < 0.05), but it did not affect the other carcass and meat traits. The optimal SID Lys requirement of 20−40 kg, 40−60 kg, and 60−90 kg of Baqing pigs fed corn−soybean meal-based diets is estimated to be 0.92%, 0.66%, and 0.55% of the diets by the quadratic curve models, respectively.

[Establishment of mouse lipogenic inflammation-mediated hepatocarcinoma model by hydrodynamic injection system].

Objective To establish and evaluate a murine lipogenic inflammation-mediated hepatocarcinoma model with hydrodynamic transfection of protein kinase B (AKT), ß-catenin, and Sleeping Beauty transposon plasmids into hepatocytes through the tail vein. Methods Isotonic physiological solution (20 µg myrAKT, 20 µg δN90-ß-catenin and 4 µg Sleeping Beauty transposase were dissolved in 2 mL of normal saline) was injected into C57BL/6 male mice aged 6-8 weeks via the tail vein in a short period of time (5-9 s) by hydrodynamic transfection, while the control plasmid was injected with the same method. Mice were then sacrificed 8, 12, or 16 weeks after the injection of these plasmids, and their serum and liver tissues were collected. RNA samples were extracted for further analysis. HE staining was used to detect liver tissue structure and inflammatory cell infiltration; oil red O staining was used to examine lipid deposition in liver cells; sirius red and Masson staining were used to identify collagen deposition in liver tissues; immunohistochemistry was performed to evaluate the proliferation abilities of the cells and microvascular density and distribution; real-time quantitative PCR was used to reflect the expression of tumor-associated genes; flow cytometry was used for immune cell phenotype identification. Results Sixteen weeks after hydrodynamic transfection, 100% (20/20) mice developed liver tumors in the experimental group. Compared with the control group, the body mass of mice in the experimental group increased slowly and liver mass increased significantly. HE staining showed that a large number of hepatocytes appeared to be steatosis, inflammatory cell infiltration increased, liver tumor cells were round or irregular-shaped, the cytoplasmic ratio was imbalanced, tumor cell atypia was obvious and tumor cells were distributed in clustering; ki67 positive cells were enhanced. Serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), γ-glutamyl transpeptidase (γ-GT), and alkaline phosphatase (ALP) levels remarkably increased. AFP mRNA also significantly increased in liver tissue. CD31 staining showed increased vessel density and vessel structure disorganization. A large number of inflammatory cells, especially myeloid and lymphoid cells, were infiltrated into the liver tissue. Conclusion Murine lipogenic inflammation-mediated hepatocarcinoma model that is caused by non-resolving inflammation was successfully established by hydrodynamic transfection with AKT and ß-catenin plasmids. This method is easier performed and reproducible, the induction time is relatively short with a high rate of tumor formation.

Dietary Supplementation with Phytase and Protease Improves Growth Performance, Serum Metabolism Status, and Intestinal Digestive Enzyme Activities in Meat Ducks.

Two experiments were conducted to investigate the effects of dietary supplementation with protease and phytase on growth performance, serum physiochemical parameters, and activities of digestive enzymes in jejunal digesta of meat ducks. Experiment 1 was carried out to determine the effects of different protease or phytase on growth performance, serum physiochemical parameter, and activities of digestive enzymes in jejunal digesta of meat ducks to select the optimal phytase or protease. According to the hatching age and initial weight, a total of 5040 Cherry Valley ducks (15 days of age) were randomly assigned into six treatments. Treatments included a basal control diet (CON) and 5 basal diets supplemented with different enzyme preparations, which were phytase preparation A (PA, 160 g/t), phytase preparation B (PB, 800 g/t), protease preparation A (PTA, 80 g/t), protease preparation B (PTB, 300 g/t) and protease preparation C (PTC, 200 g/t). The enzyme activities were as follows: Phytase A and B as well as protease A, B, and C were 50,000, 10,000, 250,000, 50,000, and 60,000 U/g, respectively. Each treatment had 7 replicates with 120 meat ducks per replicate. Experiment 1 lasted for 28 days. The results showed that: compared with the CON group, the PA group significantly decreased contents of serum phosphorus and calcium (p < 0.05), and the PTA, PTB, and PTC groups had higher activities of trypsin in jejunal digesta (p < 0.05), and the activity of jejunal chymotrypsin in PTA group was greater (p < 0.05). Experiment 2 was carried out to determine the effects of dietary supplementation with protease and phytase in low-energy and low-protein diet on growth performance, serum physiochemical parameters, and activities of digestive enzymes in jejunal digesta of meat ducks. According to the hatching age and initial weight, a total of 5760 Cherry Valley ducks (15 days of age) were randomly assigned into four treatments on the basis of a trial of 2 × 2 factorial design. Treatments included a basal control diet (PC), basal diet supplemented with enzymes (PCE), low-energy and low-protein diet (LEP), and low-energy and low-protein diet supplemented with enzymes (LEPE), the nutrient levels of energy and CP of basal diet were 2747.2 cal·ME/kg and 16.80%, respectively, and the nutrient levels of energy and CP of low-energy and low-protein diet decreased 45.90 kcal·ME/kg and 0.52% on the basis of basal diet, respectively. According to the results of experiment 1, phytase A and protease A were determined as the optimal enzyme combination of Experiment 2, and additional dosage of which were identical with Experiment 1. Each treatment had 6 replicates with 240 meat ducks per replicate. Experiment 2 lasted for 28 days. The results showed that: compared with PC and LEP groups, PCE and LEPE groups had higher final weight and average daily gain (ADG) (p < 0.05), higher activities of trypsin and chymotrypsin in jejunal digesta (p < 0.05), lower contents of serum calcium and phosphorus as well as higher levels of high-density lipoprotein in the serum (p < 0.05). In conclusion, dietary supplementation with phytase and protease in different energy and protein diets could increase digestive enzymes in jejunal digesta, effect serum physiochemical parameters, improve metabolic status, and increase the growth performance of meat ducks. Meanwhile, with the dietary supplementation with phytase and protease in the lower energy and protein diet, the growth performance could reach to the degree of the higher energy and increased protein diet, but without the addition of phytase and protease.

circRNA Profiling Reveals an Abundant circFUT10 that Promotes Adipocyte Proliferation and Inhibits Adipocyte Differentiation via Sponging let-7.

Adipose development is regulated by a series of complex processes, and non-coding RNAs (ncRNAs), including circular RNAs (circRNAs), play important roles in regulating proliferation and differentiation of adipocytes. In this study, we profiled circRNA expression in cattle fat tissue during calf and adult developmental stages and detected 14,274 circRNA candidates. Some circRNAs are differentially expressed between two developmental stages. We characterized circFUT10, named for its host gene FUT10, a highly expressed and abundant circRNA. Luciferase screening, an RNA-binding protein immunoprecipitation (RIP) assay, quantitative real-time PCR, and western blotting assays indicated that circFUT10 directly binds let-7c/let-e, and PPARGC1B (peroxisome proliferator-activated receptor γ coactivator 1-ß) is identified as a target of let-7c. Flow cytometry, EdU (5-ethynyl-2'-deoxyuridine) incorporation, a CCK-8 (cell counting kit-8) assay, oil red O staining, and western blotting assays demonstrated that circFUT10 promotes adipocyte proliferation and inhibits cell differentiation by sponging let-7c. The results demonstrate that circFUT10 binding of let-7c promotes cell proliferation and inhibits cell differentiation by targeting PPARGC1B in cattle adipocytes.