Elevated extracellular calcium ions accelerate the proliferation and migration of HepG2 cells and decrease cisplatin sensitivity.

Hepatoblastoma is the most frequent liver malignancy in children. HepG2 has been discovered as a hepatoblastoma-derived cell line and tends to form clumps in culture. Intriguingly, we observed that the addition of calcium ions reduced cell clumping and disassociated HepG2 cells. The calcium signal is in connection with a series of processes critical in the tumorigenesis. Here, we demonstrated that extracellular calcium ions induced morphological changes and enhanced the epithelial-mesenchymal transition in HepG2 cells. Mechanistically, calcium ions promoted HepG2 proliferation and migration by up-regulating the phosphorylation levels of focal adhesion kinase (FAK), protein kinase B, and p38 mitogen-activated protein kinase. The inhibitor of FAK or Ca 2+/calmodulin-dependent kinase Ⅱ (CaMKⅡ) reversed the Ca 2+-induced effects on HepG2 cells, including cell proliferation and migration, epithelial-mesenchymal transition protein expression levels, and phosphorylation levels of FAK and protein kinase B. Moreover, calcium ions decreased HepG2 cells' sensitivity to cisplatin. Furthermore, we found that the expression levels of FAK and CaMKⅡ were increased in hepatoblastoma. The group with high expression levels of FAK and CaMKⅡ exhibited significantly lower ImmunoScore as well as CD8 + T and NK cells. The expression of CaMKⅡ was positively correlated with that of PDCD1 and LAG3. Correspondingly, the expression of FAK was negatively correlated with that of TNFSF9, TNFRSF4, and TNFRSF18. Collectively, extracellular calcium accelerates HepG2 cell proliferation and migration via FAK and CaMKⅡ and enhances cisplatin resistance. FAK and CaMKⅡ shape immune cell infiltration and responses in tumor microenvironments, thereby serving as potential targets for hepatoblastoma.

Deletion of Brucella transcriptional regulator GntR10 regulated the expression of quorum sensing system and type IV secretion system effectors, which affected the activation of NF-κB.

GntR10 is a transcriptional regulator in Brucella. Nuclear factor-kappa B (NF-κB) is involved in many cellular activities, playing major roles in orchestrating the expression of inflammatory genes and regulating protein function that is essential for pathogenic bacteria during infection. GntR10 deletion was previously found to affect the growth and the virulence of Brucella and expression levels of target genes of GntR10 in mice. However, the mechanisms of affection of NF-κB regulated by Brucella GntR10 are still unclear. Here, GntR10 deletion could regulate the expression of LuxR-type transcriptional activators (VjbR and BlxR) of the quorum sensing system (QSS) and type IV secretion system (T4SS) effectors (BspE and BspF) of Brucella. It could further inhibit the activation of the regulator NF-κB and affect the virulence of Brucella. This research provides new insights into the designing of Brucella vaccines and the screening of drug targets. SIGNIFICANCE: Transcriptional regulators are predominant bacterial signal transduction factors. The pathogenicity of Brucella is due to its ability to regulate the expression of virulence related genes including quorum sensing system (QSS) and type IV secretion system (T4SS). Transcriptional regulators are designed to regulate gene expression and enact an appropriate adaptive physiological response. Here, we show that Brucella transcriptional regulator GntR10 regulated the expression of QSS and T4SS effectors, which affected the activation of NF-κB.

Intestinal segment and vitamin D3 concentration affect gene expression levels of calcium and phosphorus transporters in broiler chickens.

Two experiments were conducted in this research. Experiment 1 investigated the spatial expression characteristics of calcium (Ca) and phosphorus (P) transporters in the duodenum, jejunum, and ileum of 21-day-old broilers provided with adequate nutrient feed. Experiment 2 evaluated the effects of dietary vitamin D3 (VD3) concentration (0, 125, 250, 500, 1,000, and 2,000 IU/kg) on growth performance, bone development, and gene expression levels of intestinal Ca and P transporters in 1-21-day-old broilers provided with the negative control diet without supplemental VD3. Results in experiment 1 showed that the mRNA levels of calcium-binding protein 28-kDa (CaBP-D28k), sodium-calcium exchanger 1 (NCX1), plasma membrane calcium ATPase 1b (PMCA1b), and IIb sodium-phosphate cotransporter (NaPi-IIb) were the highest in the broiler duodenum. By contrast, the mRNA levels of inorganic phosphate transporter 1 (PiT-1) and 2 (PiT-2) were the highest in the ileum. Results in experiment 2 showed that adding 125 IU/kg VD3 increased body weight gain (BWG), feed intake (FI), bone weight, and percentage and weight of Ca and P in the tibia and femur of 1-21-day-old broilers compared with the negative control diet (p < 0.05). The rise in dietary VD3 levels from 125 to 1,000 IU/kg further increased the BWG, FI, and weights of the bone, ash, Ca, and P (p < 0.05). No difference in growth rate and leg bone quality was noted in the broilers provided with 1,000 and 2,000 IU/kg VD3 (p > 0.05). Supplementation with 125-2,000 IU/kg VD3 increased the mRNA abundances of intestinal Ca and P transporters to varying degrees. The mRNA level of CaBP-D28k increased by 536, 1,161, and 28 folds in the duodenum, jejunum, and ileum, respectively, after adding 1,000 IU/kg VD3. The mRNA levels of other Ca and P transporters (PMCA1b, NCX1, NaPi-IIb, PiT-1, and PiT-2) increased by 0.57-1.74 folds by adding 1,000-2,000 IU/kg VD3. These data suggest that intestinal Ca and P transporters are mainly expressed in the duodenum of broilers. Moreover, the addition of VD3 stimulates the two mineral transporter transcription in broiler intestines.

Correlation between stem cell molecular phenotype and atherosclerotic plaque neointima formation and analysis of stem cell signal pathways.

Vascular stem cells exist in the three-layer structure of blood vessel walls and play an indispensable role in angiogenesis under physiological conditions and vascular remodeling under pathological conditions. Vascular stem cells are mostly quiescent, but can be activated in response to injury and participate in endothelial repair and neointima formation. Extensive studies have demonstrated the differentiation potential of stem/progenitor cells to repair endothelium and participate in neointima formation during vascular remodeling. The stem cell population has markers on the surface of the cells that can be used to identify this cell population. The main positive markers include Stem cell antigen-1 (Sca1), Sry-box transcription factor 10 (SOX10). Stromal cell antigen 1 (Stro-1) and Stem cell growth factor receptor kit (c-kit) are still controversial. Different parts of the vessel have different stem cell populations and multiple markers. In this review, we trace the role of vascular stem/progenitor cells in the progression of atherosclerosis and neointima formation, focusing on the expression of stem cell molecular markers that occur during neointima formation and vascular repair, as well as the molecular phenotypic changes that occur during differentiation of different stem cell types. To explore the correlation between stem cell molecular markers and atherosclerotic diseases and neointima formation, summarize the differential changes of molecular phenotype during the differentiation of stem cells into smooth muscle cells and endothelial cells, and further analyze the signaling pathways and molecular mechanisms of stem cells expressing different positive markers participating in intima formation and vascular repair. Summarizing the limitations of stem cells in the prevention and treatment of atherosclerotic diseases and the pressing issues that need to be addressed, we provide a feasible scheme for studying the signaling pathways of vascular stem cells involved in vascular diseases.

Progress in Multidisciplinary Treatment of Fournier's Gangrene.

Fournier's gangrene (FG) is a life-threatening and special form of necrotizing fasciitis, characterized by occult onset, rapid progress and high mortality, occurring mainly in men over 50 years of age. Risk factors of FG include diabetes, HIV infection, chronic alcoholism and other immunosuppressive state. FG was previously considered as an idiopathic disease, but in fact, three quarters of the infections originated from the skin, urethra and gastrointestinal tract. Initial symptoms of FG are often inconsistent with severity and can progress promptly to fatal infection. Although the treatment measures of FG have been improved in recent years, the mortality does not seem to have decreased significantly and remains at 20% - 30%. The time to identify FG and the waiting period before surgical debridement are directly related to the prognosis. Therefore, in addition to the combination of intensive fluid resuscitation and broad-spectrum antibiotics, treatment of FG should particularly emphasize the importance of early surgical debridement assisted with fecal diversion and skin reconstruction when necessary. This paper is to briefly summarize the progress in the definition, epidemiology, clinical manifestations, diagnosis, treatment and prognosis of Fournier's gangrene in recent years, more importantly, illustrates the importance of multidisciplinary cooperation in the management of FG.

1,25-Dihydroxycholecalciferol Improved the Growth Performance and Upregulated the Calcium Transporter Gene Expression Levels in the Small Intestine of Broiler Chickens.

1,25-Dihydroxycholecalciferol (1,25-(OH)2-D3) is the final active product of vitamin D. This study aimed to investigate the effects of 1,25-(OH)2-D3 on growth performance, bone development, and calcium (Ca) transporter gene expression levels in the small intestine of broiler chickens. On the day of hatching, 140 female Ross 308 broilers were randomly allotted into two treatments with five replicates (14 birds per replicate). Two levels of 1,25-(OH)2-D3 (0 and 1.25 µg/kg) were added to the basal diet without vitamin D. Results showed that the addition of 1.25 µg/kg 1,25-(OH)2-D3 increased the average daily feed intake and the average daily gain and decreased the feed conversion ratio and mortality in 1- to 19-day-old broiler chickens compared with the basal diet without vitamin D (P<0.05). 1,25-(OH)2-D3 also enhanced the length, weight, ash weight, and the percentage contents of ash, Ca, and P in the tibia and femur of broilers (P<0.05). The mRNA expression levels of the Ca-binding protein (CaBP-D28k) in the duodenum, jejunum, and ileum of 19-day-old broilers increased to 88.1-, 109.1-, and 2.7-fold, respectively, after adding 1,25-(OH)2-D3 (P<0.05). The mRNA expression levels of the plasma membrane Ca ATPase 1b (PMCAlb) in the duodenum and the sodium (Na)/ Ca exchanger 1 (NCX1) in the duodenum and the jejunum were also enhanced to 1.57-2.86 times with the addition of 1,25-(OH)2-D3 (P<0.05). In contrast, the mRNA expression levels of PMCA1b and NCX1 in the ileum and that of vitamin D receptor (VDR) in the small intestine were not affected by 1,25-(OH)2-D3 (P>0.05). These data indicate that 1,25-(OH)2-D3 upregulated Ca transporter gene transcription and promoted Ca2+ absorption in the small intestine, especially in the proximal intestine (duodenum and jejunum), thereby improving growth performance and bone mineralization in broiler chickens.

Functional insights into Brucella transcriptional regulator ArsR.

ArsR-family transcriptional factors regulates diverse physiological functions necessary for Brucella adaptation to environmental changes. However, whether the ArsR-family transcriptional regulator are related to virulence, and the precise determination of ArsR direct targets in Brucella are still unknown. Therefore, we created a 2308ΔArsR6 mutant of B. abortus 2308 (S2308). Virulence assay was performed using a murine macrophage cell line (RAW 264.7). We performed chromatin immunoprecipitation of ArsR6 followed by next-generation sequencing (ChIP-seq). We also selected the target gene pobA (BAB2_0600), and created the mutant (2308ΔpobA). The survival capability of 2308ΔpobA strain in RAW 264.7 was detected and the levels of tumor necrosis factor-α (TNF-α), interferon-gamma (IFN-γ), interleukin-12 (IL-12) and interleukin-18 (IL-18) were also measured. The results showed that 2308ΔArsR6 reduced survival capability in RAW 264.7. We detected 40 intergenic ChIP-seq peaks of ArsR6 binding distributed across the Brucella genome. 2308ΔpobA was significantly reduced survival capability in RAW 264.7. After the macrophages were infected with 2308ΔpobA, the levels of TNF-α, IFN-γ, IL-12 and IL-18 were decreased and were significantly lower than that for the S2308-infected group, indicating that the 2308ΔpobA could reduce the secretion of inflammatory cytokines. Taken together, the research provided new insights into the functionality of ArsR6 and great significance to clarify the function of ArsR6.

Encapsulated ruthenium nanoparticles activated few-layer carbon frameworks as high robust oxygen evolution electrocatalysts in acidic media.

Noble metals have been extensively employed as high active catalysts for oxygen evolution reaction (OER), are usually subjected to serious surface transformation and poor structural stability, especially in acid media, which need imperatively remedied. Herein, the interfacial engineering of Ru via few-layer carbon (Ru@FLC) was carried out, in which FLC can significantly suppress the corrosion of Ru in acid media, ensuring the efficient interfacial charge transport between Ru and FLC. As a result, a low overpotentials@10 mA cm-2 of 258 mV and small Tafel slopes of 53.1 mV dec-1 for oxygen evolution OER were achieved in acid media. DFT calculations disclose that outer FLC could induce charge redistribution and effectively optimize intermediates free energy adsorption, resulting in greatly reduce the energy barrier for OER. Our work may offer a new avenue to produce progressive OER electrocatalysts for energy-related applications in acid solution.

Characterization of the bacterial microbiota composition and evolution at different intestinal tract in wild pigs (Sus scrofa ussuricus).

Commensal microorganisms are essential to the normal development and function of many aspects of animal biology, including digestion, nutrient absorption, immunological development, behaviors, and evolution. The specific microbial composition and evolution of the intestinal tracts of wild pigs remain poorly characterized. This study therefore sought to assess the composition, distribution, and evolution of the intestinal microbiome of wild pigs. For these analyses, 16S rRNA V3-V4 regions from five gut sections prepared from each of three wild sows were sequenced to detect the microbiome composition. These analyses revealed the presence of 6,513 operational taxonomic units (OTUs) mostly distributed across 17 phyla and 163 genera in these samples, with Firmicutes and Actinobacteria being the most prevalent phyla of microbes present in cecum and jejunum samples, respectively. Moreover, the abundance of Actinobacteria in wild pigs was higher than that in domestic pigs. At the genus level the Bifidobacterium and Allobaculum species of microbes were most abundant in all tested gut sections, with higher relative abundance in wild pigs relative to domestic pigs, indicating that in the process of pig evolution, the intestinal microbes also evolved, and changes in the intestinal microbial diversity could have been one of the evolutionary forces of pigs. Intestinal microbial functional analyses also revealed the microbes present in the small intestine (duodenum, jejunum, and ileum) and large intestine (cecum and colon) of wild pigs to engage distinct metabolic spatial structures and pathways relative to one another. Overall, these results offer unique insights that would help to advance the current understanding of how the intestinal microbes interact with the host and affect the evolution of pigs.

Broadening AIEgen application: rapid and portable sensing of foodstuff hazards in deep-frying oil.

Development of fluorescent probes that can monitor foodstuff hazards is highly desirable. Herein we report the first example of an AIEgen probe (QM-TPA), conjugated by a quinoline-malononitrile (QM) scaffold and triphenylamine unit, for direct sensing of triacylglycerol polymers in frying oil, enabling a rapid probing, on-site analysis, and portable operation in food inspection applications.

Determination and prediction of the digestible and metabolizable energy contents of corn germ meal in growing pigs.

OBJECTIVE: This experiment was conducted to determine the chemical composition, digestible energy (DE) and metabolizable energy (ME) contents of corn germ meals (CGM) and to develop equations to predict the corresponding energy contents based on the chemical characteristics of individual CGM. METHODS: Sixty-six barrows (initial body weight = 51.3±4.6 kg) were allotted to 11 diets including a basal diet and 10 CGM test diets in a completely randomized design. In the test diets, CGM was included in replacement of 30% of the energy-providing ingredients in the basal diet, resulting in a final inclusion rate of 29.1%. Each diet was fed to 6 barrows housed in individual metabolism crates for a 7-d acclimation period followed by a 5-d total but separate collection of feces and urine. RESULTS: Considerable variation was observed in acid-hydrolyzed ether extract, ether extract, ash, calcium (Ca) and total phosphorus contents among the CGM samples. On dry matter (DM) basis, the DE and ME contents of the CGM ranged from 10.22 to 15.83 MJ/kg and from 9.94 to 15.43 MJ/kg, respectively. The acid detergent fiber (ADF) contents were negatively correlated with the DE and ME contents of CGM samples. The best-fit prediction equations for the DE and ME values (MJ/kg DM) of the 10 CGM were: DE = 26.85-0.28 insoluble dietary fiber (%)-17.79 Ca (%); ME = 21.05-0.43 ADF (%)-11.40 Ca (%). CONCLUSION: The chemical compositions of CGM vary depending on sources, particularly in ether extract and Ca. The DE and ME values of CGM can be predicted based on their chemical composition in growing pigs.

Effects of supplementary amino acids on available energy of soybean meal determined by difference and regression methods fed to growing pigs.

This study was conducted to evaluate effects of inclusion level and supplementary amino acids (AAs) on digestible energy (DE) and metabolizable energy (ME) values of soybean meal (SBM) fed to growing pigs, determined by difference and regression methods. Sixty pigs were fed 10 diets according to a 5 × 2 factorial arrangement. Two control diets contained 97.34% corn without supplementary AAs or 95.61% corn with supplementary AAs. Eight diets were formulated by replacing corn and AAs in control diets with 8%, 15%, 25% and 31% SBM. There was no difference in DE and ME values of SBM determined by difference method as inclusion level of SBM increased or crystalline AAs were added. No difference was observed in DE and ME values of SBM determined by the two methods in diets without supplementary AAs, but the values determined by the difference method were greater (P < 0.05) than those determined by regression method when crystalline AAs were added in diets. In conclusion, inclusion level and supplementary AAs did not affect DE and ME values of SBM calculated by difference or regression methods. There were differences in DE and ME values of SBM determined by the difference method and the regression method when crystalline AAs were added in diets.

The Effect of Inclusion Level of Soybean Oil and Palm Oil on Their Digestible and Metabolizable Energy Content Determined with the Difference and Regression Method When Fed to Growing Pigs.

This experiment was conducted to determine the effects of inclusion level of soybean oil (SO) and palm oil (PO) on their digestible and metabolism energy (DE and ME) contents when fed to growing pigs by difference and regression method. Sixty-six crossbred growing barrows (Duroc×Landrace×Yorkshire and weighing 38.1±2.4 kg) were randomly allotted to a 2×5 factorial arrangement involving 2 lipid sources (SO and PO), and 5 levels of lipid (2%, 4%, 6%, 8%, and 10%) as well as a basal diet composed of corn and soybean meal. The barrows were housed in individual metabolism crates to facilitate separate collection of feces and urine, and were fed the assigned test diets at 4% of initial body weight per day. A 5-d total collection of feces and urine followed a 7-d diet adaptation period. The results showed that the DE and ME contents of SO and PO determined by the difference method were not affected by inclusion level. The DE and ME determined by the regression method for SO were greater compared with the corresponding respective values for PO (DE: 37.07, ME: 36.79 MJ/kg for SO; DE: 34.11, ME: 33.84 MJ/kg for PO, respectively). These values were close to the DE and ME values determined by the difference method at the 10% inclusion level (DE: 37.31, ME: 36.83 MJ/kg for SO; DE: 34.62, ME: 33.47 MJ/kg for PO, respectively). A similar response for the apparent total tract digestibility of acid-hydrolyzed ether extract (AEE) in lipids was observed. The true total tract digestibility of AEE in SO was significantly (p<0.05) greater than that for PO (97.5% and 91.1%, respectively). In conclusion, the DE and ME contents of lipid was not affected by its inclusion level. The difference method can substitute the regression method to determine the DE and ME contents in lipids when the inclusion level is 10%.

Estimation of the optimal standardized ileal digestible lysine requirement for primiparous lactating sows fed diets supplemented with crystalline amino acids.

This experiment was conducted to determine the optimal standardized ileal digestible lysine (SID Lys) level in diets fed to primiparous sows during lactation. A total of 150 (Landrace × Large White) crossbred gilts (weighing 211.1 ± 3.5 kg with a litter size of 11.1 ± 0.2) were fed lactation diets (3325 kcal metabolizable energy (ME)/kg) containing SID Lys levels of 0.76, 0.84, 0.94, 1.04 or 1.14%, through 28 days lactation. Gilts were allocated to treatments based on their body weight and backfat thickness 48 h after farrowing. Gilt body weight loss was significantly (P < 0.05) decreased by increasing dietary SID Lys levels. Fitted broken-line (P < 0.05) and quadratic plot (P < 0.05) analysis of body weight loss indicated that the optimal SID Lys for primiparous sows was 0.85 and 1.01%, respectively. Average daily feed intake (ADFI), weaning-to-estrus interval and subsequent conception rate were not affected by dietary SID Lys levels. Increasing dietary lysine had no effect on litter performances. Protein content in milk was increased by dietary SID Lys (P < 0.05). Dietary SID Lys tended to increase concentrations of serum insulin-like growth factor I (P = 0.066). These results of this experiment indicate that the optimal dietary SID Lys for lactating gilts was at least 0.85%, which approaches the recommendation of 0.84% that is estimated by the National Research Council (2012).

Effect of variety and drying method on the nutritive value of corn for growing pigs.

BACKGROUND: This experiment was conducted to determine the nutritive value of corn from the north of China for growing pigs. The experiment examined corn variety (LS1, LS2, LS3 and LS4) grown in one location, drying method (sun dried and artificially dried) and different drying temperatures. Corn harvested at 20-25% moisture was dried to about 12% moisture by sun drying and artificially drying at 80, 100, or 120°C in a fluidized bed dryer. Ninety-six barrows (average BW of 33.4 ± 2.7 kg) were housed in individual metabolism crates to facilitate separate collection of feces and urine. A five-day collection period followed a seven-day diet acclimation period. RESULTS: The results indicated that variety significantly influenced (P < 0.01) the 1,000 kernel weight of corn but not the bulk weight. Variety also influenced the available energy content (digestible energy of dry matter, P < 0.01; metabolisable energy of dry matter, P < 0.01) and digestibility of organic matter (P < 0.01), as well as dry matter (P < 0.01) and gross energy (GE) content (P < 0.02). The drying method of corn significantly influenced the 1,000 kernel weight (P < 0.01), bulk weight (P < 0.01) and digestibility of ether extract (EE) (P < 0.01). No effect of drying temperature on the digestibility of organic matter, dry matter (DM), crude protein (CP), neutral detergent fiber (NDF), acid detergent fiber (ADF) and gross energy was observed, but gelatinization (P < 0.05) and test weight (P < 0.01) decreased with an increase in temperature. CONCLUSIONS: Variety has a significant impact on the nutritive value of corn for growing pigs, and greater attention needs to be paid to these influences in the assignment of the nutritive value of corn given to growing pigs.