Cinnamaldehyde affects lipid droplets metabolism after adipogenic differentiation of C2C12 cells.

BACKGROUND: Based on our previous research conducted on cinnamaldehyde (CA) exhibiting its ability to improve the growth performance of fattening pigs and the adipogenesis induction model of C2C12 cells constructed in our laboratory, we explored the effects of CA on the generation and development of lipid droplets (LDs) in adipogenic differentiated C2C12 cells. METHODS AND RESULTS: C2C12 cells were treated with either 0.4 mM or 0.8 mM CA. BODIPY staining and triglyceride measurements were conducted to observe the morphology of LDs, and Western blotting was used to measure the expression of their metabolism-related proteins. The results showed that the average number of LDs in the CA treatment groups was more than the control group (P < 0.05), whereas the average LD size and triglyceride content decreased (P < 0.05). Compared with the control group, the expression levels of fusion-related genes in the LDs of the CA treatment group significantly decreased, while decomposition-related genes and autophagy-related genes in the LDs in C2C12 cells significantly increased (P < 0.01). CONCLUSION: Cinnamaldehyde promoted the decomposition and autophagy of lipid droplets in C2C12 cells and inhibited the fusion of lipid droplets.

Confirmation and variability of the Allee effect inDictyostelium discoideumcell populations, possible role of chemical signaling within cell clusters.

In studies of the unicellular eukaryoteDictyostelium discoideum, many have anecdotally observed that cell dilution below a certain 'threshold density' causes cells to undergo a period of slow growth (lag). However, little is documented about the slow growth phase and the reason for different growth dynamics below and above this threshold density. In this paper, we extend and correct our earlier work to report an extensive set of experiments, including the use of new cell counting technology, that set this slow-to-fast growth transition on a much firmer biological basis. We show that dilution below a certain density (around 104cells ml-1) causes cells to grow slower on average and exhibit a large degree of variability: sometimes a sample does not lag at all, while sometimes it takes many moderate density cell cycle times to recover back to fast growth. We perform conditioned media experiments to demonstrate that a chemical signal mediates this endogenous phenomenon. Finally, we argue that while simple models involving fluid transport of signal molecules or cluster-based signaling explain typical behavior, they do not capture the high degree of variability between samples but nevertheless favor an intra-cluster mechanism.

Porcine interleukin-6 enhances the expression of CYP2C33 through a constitutive androstane receptor/retinoid X receptor-mediated pathway.

Cytochrome P450, which is expressed in humans and other animals, is a superfamily of drug-metabolizing enzymes that play important roles in the metabolism of endogenous and xenobiotic substrates via oxidation, peroxidation and reduction. Of endogenous substrates, interleukin (IL)-6 is a crucial cytokine involved in inflammation in the liver. The present study aims to elucidate the mechanisms through which IL-6 modulates cytochrome P450 expression. CYP2C33 expression was found to be increased in HepLi cells and primary porcine hepatocytes treated with IL-6 in a concentration-dependent manner. IL-6 treatment also increased the expression of the transcriptional regulators, constitutive androstane receptor (CAR) and pregnane X receptor. Overexpression of CAR promoted CYP2C33 expression at the mRNA and protein levels, whereas knockdown of CAR by small interfering RNA reduced CYP2C33 expression. Luciferase assays showed that IL-6 treatment of HepLi cells and primary porcine hepatocytes increased CYP2C33 promoter activity. Co-immunoprecipitation and western blotting demonstrated that CAR and RXR could form heterodimers. IL-6 affects CYP2C33 expression through CAR/retinoid X receptor (RXR) heterodimers.

IFN-γ regulates cytochrome 3A29 through pregnane X receptor in pigs.

1. The expression and the activity of cytochromes P450 (CYPs) can be elevated by the activation of nuclear receptors. The pregnane X receptor (PXR, or nuclear receptor NR1I2) is a ligand-activated transcription factor that mediates responses to diverse xenobiotics and endogenous chemicals. Here we investigated the regulatory role of PXR in IFN-γ-mediated CYP3A29 expression in pig liver microsomes, primary porcine hepatocytes, and a cultured hepatocyte cell line. 2. IFN-γ significantly up-regulated CYP3A29 and PXR expressions at mRNA and protein levels in a dose-dependent manner. IFN-γ treatment significantly increased the metabolism of nifedipine. PXR and IFN-γ treatments significantly enhanced the activity of CYP3A29 promoter and the upstream region from -1473 to -1021 of CYP3A29 might be PXR-binding site. Moreover, the IFN-γ-induced CYP3A29 expression was blocked by PXR knockdown, whereas CYP3A29 mRNA and protein expression levels were dramatically elevated by PXR overexpression. 3. The regulatory effect of IFN-γ on CYP3A29 expression is mediated via PXR.

Pregnane X receptor is required for IFN-α-mediated CYP3A29 expression in pigs.

Pregnane X receptor (PXR) has been identified as a central mediator for coordinate responses to xenobiotic and drug metabolism, and is the major transcriptional regulator of cytochrome P-450 (CYP). Interferon (IFN)-α is known to induce antiviral mechanisms and exert immune regulatory capacity in various cell types. Here, we used primary porcine hepatocytes and a cultured hepatocyte cell line to identify the metabolic role of PXR in IFN-α-mediated CYP3A29 expression. We found that IFN-α could activate PXR in both time- and dose-dependent manners in pigs. Activation of PXR significantly increased CYP3A29 mRNA and protein expression. Meanwhile, the expression of CYP3A29 induced by IFN-α occurred after the increase of PXR expression in porcine hepatocytes. In addition, the IFN-α-induced CYP3A29 expression was blocked by PXR knockdown. The PXR-overexpressed cells (transfected with porcine PXR) increased CYP3A29 mRNA and protein expression. Furthermore, in animal experiments, we found that IFN-α increased both CYP3A29 mRNA and protein levels. Collectively, our results suggest that PXR plays an important role in IFN-α-mediated CYP3A29 expression in porcine hepatocytes.

Processed food consumption and risk of esophageal squamous cell carcinoma: A case-control study in a high risk area.

This study was conducted to investigate the association between consumption of processed foods and esophageal cancer risk. A population-based case-control study was designed. For the present study, 254 patients with esophageal squamous cell carcinoma with pathological diagnoses were selected from Yanting during 2008 and 2010 and 254 community-based controls were selected from the same area, individually matched with cases by age and sex. Data on demographic, lifestyle and dietary factors were collected using food frequency questionnaires. A conditional logistic regression model was used to estimate the odds ratio (OR) with adjustments for potential confounders. Compared to the frequency of <1 time/week, the intake frequency of >3 times/week of preserved vegetables had a significant association with esophageal cancer (OR = 5.01, 95% confidence interval [CI] 2.07, 12.17). In stratified analyses, the OR of increasing intake of preserved vegetables for esophageal cancer were 2.02 in men (95% CI 1.18, 3.48), 3.15 in women (95% CI 1.28, 7.75), 2.41 (95% CI 1.45 4.01) in the persons <65 years old and 1.28 (95% CI 0.35, 4.65) in persons ≥65 years old. Consumption of pickled vegetables was not associated significantly with esophageal cancer risk. Intake of salted meat with a frequency of ≥1 time/week meant that the OR increased to 2.57 (95%CI 1.02, 6.43), but no significant trend or association in subgroup analysis was observed. Preserved vegetable consumption was associated with increased risk of esophageal cancer, while no association was found with pickled vegetables.

[Effects of selenium and zinc on the proliferation of human esophageal cancer cell line studied by serophysiology].

OBJECTIVE: The effect of serum from rats supplemented with selenium and zinc on the proliferation of human esophageal cancer cell line Eca109 was observed by serophysiology. METHODS: Rats were randomly divided into seven groups. Eight rats in each group were fed with basic feeds (deprived of both selenium and zinc). The experimental rat groups were supplemented with selenium or zinc at low or high dosage intragastrically for 30 days Serum selenium and zinc content of rats was measured by Graphite Furnace Atomic Absorption Spectrometry (GFAAS) and Flame Atomic Absorption Spectrophotometry (FAAS). MTT assay,3H-TDR incorporation and flow cytometry were used to explore the effect of serum from different rat groups on the growth and proliferation of cancer cell line Ecal09 cells. RESULTS: (1) The content of serum zinc in the high zinc group was the highest and the content of serum zinc was the lowest in basic diet group. The content of serum selenium in high selenium and high zinc group was the highest and the content of serum selenium was the lowest in the basic diet group. (2) In comparing the growth of control cancer cell group cultured with calf serum, the growth of cancer cells cultured with the serum from high selenium and high zinc rats was inhibited in culturing for 72 h, but the growth of normal liver cells were also inhibited. The growth of cancer cells were promoted by serum from other groups. (3) Both MTT assay and 3H-TDR incorporation test showed that the DNA synthesis in cancer cells was inhibited by the serum from high selenium and high zinc group, but the DNA synthesis of normal liver cells was also inhibited by this type of serum. The result of DNA synthesis in other cell groups was closed to the control group. CONCLUSIONS: Low serum selenium and zinc might promote the growth of EC cell. Elevating the content of serum selenium and zinc by increasing selenium and zinc intake might inhibit EC cell proliferation.

Effects of imbalance of mineral elements on peripheral neutrophil metabolism in sheep.

The imbalance of mineral element homeostasis in animals is common, causing animal immune dysfunction. Ten female sheep were randomly selected and injected with 4% (w/v) Na2EDTA through a central venous catheter to establish the mineral element imbalance model, then divided into control group (before injection) and Ethylene Diamine Tetraacetic Acid (EDTA) group (after injection). Isolation of peripheral blood neutrophils for mineral elements content determination was done using Inductively coupled plasma-mass spectrometry (ICP-MS) and nontargeted metabolomics analysis using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The ICP-MS results showed that Hg and Cs levels in neutrophils were significantly lower after injection than before (P < 0.05), but had no significant effects on other elements. Our previous study showed that serum Zn, P, K, and other 11 elements were significantly lower after the injection of Na2EDTA than before. LC-MS/MS results showed that differential metabolites are mainly involved in amino acid metabolism, lipid metabolism, and nucleotide metabolism; monoamine metabolism was weakened; and polyamine metabolism was enhanced. Under positive and negative ion modes, the mineral elements P, K, Ca, Mn, Cu, and Zn had the highest correlation with the differential metabolites of neutrophils, followed by Se, and the correlation between each mineral element and different differential metabolites was also different. The results indicated that the imbalance in mineral elements affected the metabolism of sheep neutrophils, these may affect neutrophil function, and Na2EDTA could help to reduce the level of heavy metals in the body of sheep. Our data may provide a theoretical basis for the precise regulation of animal immune metabolism by modern animal husbandry nutrition.

Effects of Fermented Bamboo Powder Supplementation on Serum Biochemical Parameters, Immune Indices, and Fecal Microbial Composition in Growing-Finishing Pigs.

This experiment aimed to investigate the effects of fermented bamboo powder (FBP) on the growth performance, serum biochemical parameters, immunoglobulins and inflammatory cytokines, and fecal microbial composition of growing−finishing pigs. A total of 108 barrows (initial body weight, 56.30 ± 0.55 kg) were randomly allocated to three dietary treatments in a 75 d trial, including a control (CON) diet and two FBP supplementation diets. The CON diet was formulated to three-phase diets according to the body weight of pigs, and the FBP diets were formulated used 5.00% (FBP1) or 10.00% (FBP2) FBP to replace the wheat bran in the CON diet, respectively. The results showed that there were no influences on growth performances between the CON diet and FBP addition diets, whereas the 5% FBP addition decreased the feed:gain of pigs compared to the pigs fed the FBP2 diet from d 0−75 (p < 0.05). Meanwhile, the FBP addition increased the high-density lipoprotein cholesterol (HDLC) and immunoglobulin A (IgA) content in serum (linear, p < 0.05), and pigs fed the FBP1 diet had greater HDLC and IgA contents in serum than those in the pigs fed the CON diet (p < 0.05). Microbial analysis showed that the FBP addition diets decreased the abundance of Spirochaetes, and the FBP2 diet increased the abundance of Firmicutes more than the CON diet (p < 0.05). In addition, the pigs fed the FBP2 diet increased the abundance of uncultured_bacterium_f_Lachnospiraceae, Ruminococcaceae_UCG-005, Prevotellaceae_UCG-003, Lachnospiraceae_XPB1014_group, and Lactobacillus more than the CON group (p < 0.05). In conclusion, the FBP supplementation to the diet had no negative effects on the growth performance and exerted beneficial effects on promoting serum biochemical and immune indices, as well as modulating the fecal microbiota of pigs. Therefore, these results showed that the fermented bamboo powder could be one potential fiber-rich ingredient for growing−finishing pigs, and that the recommended addition proportion in the growing−finishing pigs' diet is 5%.

Dietary energy sources and levels shift the multi-kingdom microbiota and functions in the rumen of lactating dairy cows.

BACKGROUND: Dietary energy source and level in lactation diets can profoundly affect milk yield and composition. Such dietary effects on lactation performance are underpinned by alteration of the rumen microbiota, of which bacteria, archaea, fungi, and protozoa may vary differently. However, few studies have examined all the four groups of rumen microbes. This study investigated the effect of both the level and source of dietary energy on rumen bacteria, archaea, fungi, and protozoa in the rumen of lactating dairy cows. A 2 × 2 factorial design resulted in four dietary treatments: low and high dietary energy levels (LE: 1.52-1.53; and HE: 1.71-1.72 Mcal/kg dry matter) and two dietary energy sources (GC: finely ground corn; and SFC: steam-flaked corn). We used a replicated 4 × 4 Latin square design using eight primiparous Chinese Holstein cows with each period lasting for 21 d. The rumen microbiota was analyzed using metataxonomics based on kingdom-specific phylogenetic markers [16S rRNA gene for bacteria and archaea, 18S rRNA gene for protozoa, and internally transcribed spacer 1 (ITS1) for fungi] followed with subsequent functional prediction using PICRUSt2. RESULTS: The GC resulted in a higher prokaryotic (bacterial and archaeal) species richness and Faith's phylogenetic diversity than SFC. For the eukaryotic (fungi and protozoa) microbiota, the LE diets led to significantly higher values of the above measurements than the HE diets. Among the major classified taxa, 23 genera across all the kingdoms differed in relative abundance between the two dietary energy levels, while only six genera (none being protozoal) were differentially abundant between the two energy sources. Based on prokaryotic amplicon sequence variants (ASVs) from all the samples, overall functional profiles predicted using PICRUSt2 differed significantly between LE and HE but not between the two energy sources. FishTaco analysis identified Ruminococcus and Coprococcus as the taxa potentially contributing to the enriched KEGG pathways for biosynthesis of amino acids and to the metabolisms of pyruvate, glycerophospholipid, and nicotinate and nicotinamide in the rumen of HE-fed cows. The co-occurrence networks were also affected by the dietary treatments, especially the LE and GC diets, resulting in distinct co-occurrence networks. Several microbial genera appeared to be strongly correlated with one or more lactation traits. CONCLUSIONS: Dietary energy level affected the overall rumen multi-kingdom microbiota while little difference was noted between ground corn and steam-flaked corn. Some genera were also affected differently by the four dietary treatments, including genera that had been shown to be correlated with lactation performance or feed efficiency. The co-occurrence patterns among the genera exclusively found for each dietary treatment may suggest possible metabolic interactions specifically affected by the dietary treatment. Some of the major taxa were positively correlated to milk properties and may potentially serve as biomarkers of one or more lactation traits.

Relationship between CYP3A29 and pregnane X receptor in landrace pigs: Pig CYP3A29 has a similar mechanism of regulation to human CYP3A4.

The objective of this study was to provide evidence of the validity of utilizing pigs as a model to study the regulation of human CYP3A4, with special emphasis on drug-drug interactions. We determined the mRNA expression and distribution of CYP3A and metabolic nuclear receptors in different tissues isolated from landrace pigs. Our results showed that CYP3A and metabolic nuclear receptor mRNAs were most highly expressed in liver tissues. The expression of the metabolic nuclear receptor pregnane X receptor (PXR) had a significant correlation with expression of CYP3A29, an analog of human CYP3A4. The correlation between their transcriptional levels was further demonstrated using LPS and TNF-α. The mRNA and protein expression of CYP3A29 and PXR in HepLi cells was significantly reduced by LPS and TNF-α treatment. CYP3A29 promoter activity was dramatically elevated by PXR over expression, whereas LPS and TNF-α treatment inhibited the enhanced CYP3A29 promoter activity that was induced by PXR; presumably through inhibition of PXR promoter activity. Furthermore, the inhibition of CYP3A29 promoter activity by LPS and TNF-α treatment was blocked by knockdown of PXR or retinoid X receptor (RXR). These data suggest high similarity in the regulation mechanism of pig CYP3A29 and human CYP3A4. Our research provided a significant evaluation to determine whether pigs are suitable as an experimental animal model.

Cecropin B Represses CYP3A29 Expression through Activation of the TLR2/4-NF-κB/PXR Signaling Pathway.

Cecropins are peptide antibiotics used as drugs and feed additives. Cecropin B can inhibit the expression of CYP3A29, but the underlying mechanisms remain unclear. The present study was designed to determine the mechanisms responsible for the effects of cecropin B on CYP3A29 expression, focusing on the Toll-like receptors (TLRs) and NF-κB pathways. Our results indicated that the CYP3A29 expression was inhibited by cecropin B, which was regulated by pregnane X receptor (PXR) in a time- and dose-dependent manner. Cecropin B-induced NF-κB activation played a pivotal role in the suppression of CYP3A29 through disrupting the association of the PXR/retinoid X receptor alpha (RXR-α) complex with DNA sequences. NF-κB p65 directly interacted with the DNA-binding domain of PXR, suppressed its expression, and inhibited its transactivation, leading to the downregulation of the PXR-regulated CYP3A29 expression. Furthermore, cecropin B activated pig liver cells by interacting with TLRs 2 and 4, which modulated NF-κB-mediated signaling pathways. In conclusion, cecropin B inhibited the expression of CYP3A29 in a TLR/NF-κB/PXR-dependent manner, which should be considered in future development of cecropins and other antimicrobial peptides.

Milk production and composition responds to dietary neutral detergent fiber and starch ratio in dairy cows.

This study was designed to investigate whether dietary neutral detergent fiber (NDF) : starch ratio could be considered as a nutritional indicator to evaluate carbohydrate composition and manipulate milk production and composition synthesis. Eight primiparous dairy cows were assigned to four total mixed rations with NDF : starch ratios of 0.86, 1.18, 1.63 and 2.34 from T1 to T4 in a replicated 4 × 4 Latin square design. Dry matter intake and milk production were decreased from T1 to T4. Digestibility of dry matter, organic matter, NDF and crude protein were linearly decreased from T1 to T4. As NDF : starch ratio increased, milk protein content and production, and milk lactose content and production were linearly reduced. However, milk fat content was linearly increased from T1 to T4. Quadratic effect was observed on milk fat production with the highest level in T3. Averaged rumen pH was linearly increased from T1 to T4, and subacute rumen acidosis occurred in T1. Ruminal propionate and butyrate concentration were linearly decreased, and microbial crude protein and metabolizable protein decreased from T1 to T4. It is concluded that NDF : starch ratio can be considered as a potential indicator to evaluate dietary carbohydrate composition and manipulate milk production and composition synthesis.

Esophageal cancer mortality during 2004-2009 in Yanting County, China.

OBJECTIVE: Yanting County is a high risk area for esophageal cancer (EC) in China. The purpose of this study was to describe the mortality and mortality change of EC from 2004 to 2009 in Yanting County. METHODS: EC mortality data from 2004 to 2009 obtained from the Cancer Registry in Yanting were analyzed. Annual percentage changes (APC) were calculated to assess the trends in EC mortality. Age-standardized mortality was calculated based on world standard population of 2000. RESULTS: The average EC mortality was 54.7/105 in males and 31.6/105 in females over the 6 years. A decline in EC mortality with time was observed in both genders, with a rate of -8.70% per year (95% CI: -13.23%~-3.93%) in females and -4.11% per year (95%CI: -11.16%~3.50%) in males. CONCLUSION: EC mortality decreased over the six years in both genders, although it remained high in the Yanting area. There is still a need to carry out studies of risk factors for improved cancer prevention and further reduction in the disease burden.

Major cancer mortality and changes in Yanting, 2004-2009: introduction to cancer challenges in a high risk area.

Yanting County is one of high risk areas of esophageal cancer in China. Recently, the economic level has improved to a new standard, but cancer issues have not been updated. This study aimed to describe the main cancer mortalities and change from 2004 to 2009 and provide an evidence base for future active strategies. Yanting Cancer Research Institute provided all cancer mortality data and age-standardized rates were calculated based on the world standard population 2000. Annual percentage change was used to estimate the time trend for each cancer. Mortality from upper gastrointestinal cancers, but not other cancers, was much higher than worldwide average figures. Rates for esophageal cancer declined over the 6 years, but lung cancer mortality showed an upward trend. For gastric and liver cancer, no obvious change was observed. Considering the high mortality from upper gastrointestinal cancers, it is necessary to take actions investigating the risk factors and addressing the issues of prevalent cancer challenges.

Relationship between genetic polymorphisms of ALDH2 and ADH1B and esophageal cancer risk: a meta-analysis.

AIM: To evaluate the contribution of alcohol dehydrogenase-1B (ADH1B) and aldehyde dehydrogenase-2 (ALDH2) polymorphisms to the risk of esophageal cancer. METHODS: Nineteen articles were included by searching MEDLINE, EMBASE and the Chinese Biomedical Database, 13 on ADH1B and 18 on ALDH2. We performed a meta-analysis of case-control studies including 13 studies on ADH1B (cases/controls: 2390/7100) and 18 studies on ALDH2 (2631/6030). RESULTS: The crude odds ratio [OR (95% confidence interval)] was 2.91 (2.04-4.14) for ADH1B*1/*1 (vs ADH1B*2/*2) and 1.32 (1.17-1.49) for ADH1B*1/*2. The crude OR for ALDH2*1/*2 (vs ALDH2*1/*1) was 2.52 (1.76-3.61). ADH1B*1/*1 increased the risk of esophageal cancer among never/rare [1.56 (0.93-2.61)], moderate [2.71 (1.37-5.35)], and heavy drinkers [3.22 (2.27-4.57)]. ADH1B*1/*2 was associated with a modest risk among moderate drinkers [1.43 (1.09-1.87)]. ALDH2*1/*2 increased the risk among never/rare [1.28 (0.91-1.80)], moderate [3.12 (1.95-5.01)], and heavy [7.12 (4.67-10.86)] drinkers, and among ex-drinkers [5.64 (1.57-20.25)]. ALDH2*2/*2 increased the risk among drinkers [4.42 (1.72-11.36)]. ADH1B*1/*1 plus ALDH2*1/*2 was associated with the highest risk for heavy drinkers [12.45 (2.9-53.46)]. The results of the meta-regression analysis showed that the effects of ADH1B*1/*1 and ALDH2*1/*2 increased with the level of alcohol consumption. ALDH2*1/*2 was associated with a high risk among Taiwan Chinese and Japanese drinkers, as opposed to a moderate risk among drinkers in high-incidence regions of Mainland China. ADH1B*1/*1 in heavy drinkers and ALDH2*1/*2 in moderate-to-heavy drinkers was associated with similarly high risk among both men and women. CONCLUSION: ADH1B/ALDH2 genotypes affect the risk of esophageal cancer, and the risk is modified by alcohol consumption, ethnicity, and gender.