TSA attenuates the progression of c-Myc-driven hepatocarcinogenesis by pAKT-ADH4 pathway.

Hepatocellular carcinoma (HCC) is the primary malignant tumor of the liver. c-Myc is one of the most common oncogenes in clinical settings, and amplified levels of c-Myc are frequently found in HCC. Histone deacetylase inhibitors (HDACi), such as Trichostatin A (TSA), hold enormous promise for the treatment of HCC. However, the potential and mechanism of TSA in the treatment of c-Myc-induced HCC are unclear. In this study, we investigated the effects of TSA treatment on a c-Myc-induced HCC model in mice. TSA treatment delayed the development of HCC, and liver function indicators such as ALT, AST, liver weight ratio, and spleen weight ratio demonstrated the effectiveness of TSA treatment. Oil red staining further demonstrated that TSA attenuated lipid accumulation in the HCC tissues of mice. Through mRNA sequencing, we identified that TSA mainly affected cell cycle and fatty acid degradation genes, with alcohol dehydrogenase 4 (ADH4) potentially being the core molecular downstream target. QPCR, immunohistochemistry, and western blot analysis revealed that ADH4 expression was repressed by c-Myc and restored after TSA treatment both in vitro and in vivo. Furthermore, we observed that the levels of total NAD+ and NADH, NAD+, NAD+/NADH, and ATP concentration increased after c-Myc transfection in liver cells but decreased after TSA intervention. The levels of phosphorylated protein kinase B (p-AKT) and p-mTOR were identified as targets regulated by TSA, and they governed the ADH4 expression and the downstream regulation of total NAD+ and NADH, NAD+, NAD+/NADH, and ATP concentration. Overall, our study suggests that TSA has a therapeutic effect on c-Myc-induced HCC through the AKT-mTOR-ADH4 pathway. These findings provide valuable insights into the potential treatment of HCC using TSA and shed light on the underlying molecular mechanisms involved.

Effect of Yucca schidigera extract on the growth performance, intestinal antioxidant status, immune response, and tight junctions of mirror carp (Cyprinus carpio).

The current study was designed to evaluate the effect of Yucca schidigera extract (YSE) on the growth performance, intestinal antioxidant status, immune response, and tight junctions of mirror carp (Cyprinus carpio). A total of 450 mirror carp (45.21 ± 0.43 g) were fed diets supplemented with 0, 200, or 400 mg/kg YSE for 8 weeks. Compared with the control (0 mg/kg), the final body weight and weight gain rate were significantly higher in the 400 mg/kg YSE group (P < 0.05), and the serum ammonia concentration was significantly lower in both YSE groups (P < 0.05). Additionally, the total antioxidant capacity was significantly higher in the 400 mg/kg YSE group (P < 0.05), and the malondialdehyde content was significantly lower in both YSE groups (P < 0.05). Complement 3 and 4 contents were significantly higher in the 400 mg/kg YSE group (P < 0.05), and lysozyme was significantly higher in both YSE groups compared to the control group (P < 0.05). The relative mRNA levels of copper zinc superoxide dismutase, catalase, glutathione peroxidase1a, and nuclear factor erythroid 2-related factor 2 as well as transforming growth factor ß were significantly higher in both YSE supplemented groups compared to the control (P < 0.05), whereas the relative mRNA level of Kelch-like ECH-associated protein 1 was significantly lower in both YSE supplemented groups (P < 0.05). The relative mRNA levels of interleukin 1ß and interleukin 6 were significantly lower in the 400 mg/kg YSE supplemented group compared to the control (P < 0.05). Additionally, both YSE levels decreased the relative mRNA expression of tumour necrosis factor-α (P < 0.05). The relative mRNA levels of ZO-1 and claudin 11 were significantly higher in both YSE supplemented groups (P < 0.05), and the relative mRNA level of occludin was significantly higher in the 200 mg/kg YSE group than the control and 400 mg/kg YSE groups (P < 0.05). In conclusion, dietary supplementation with 400 mg/kg YSE improved the growth, intestinal antioxidant status, immune response, and tight junctions of mirror carp.

Dietary guanidinoacetic acid supplementation improved carcass characteristics, meat quality and muscle fibre traits in growing-finishing gilts.

This study was designed to evaluate the effects of guanidinoacetic acid (GAA) on growth performance, carcass characteristics, meat and muscle fibre traits of growing-finishing gilts. 300 female PIC pigs were randomly divided (30.10 ± 2.94 kg) into 2 treatments with 6 replicates of 25 each for a 100-day trial. Two dietary treatments were comprised of a control diet and a control diet fortified with 450 mg/kg GAA. Growth performance was evaluated for each phase. Carcass characteristics and meat quality were determined at last phase. Gilts had free access to feed and water during the experiment. The result indicated that GAA did not affect growth performance (p > 0.05). GAA not only increased longissimus dorsi (LM) muscle weight but also decreased its shear force, b*value and drip loss (p < 0.05). Mandibular fat index was decreased by GAA (p < 0.05). GAA upregulated myosin heavy chain (MyHC) I mRNA expression with lower myofibre cross-sectional area and fibre diameter in LM muscle (p < .05). In conclusion, GAA can improve carcass characteristics and meat quality by changing muscle fibre characteristics and reducing mandibular fat index in finishing gilts.

Effects of the different levels of dietary trace elements from organic or inorganic sources on growth performance, carcass traits, meat quality, and faecal mineral excretion of broilers.

This experiment was conducted to evaluate the effects of different sources and levels of trace elements on growth performance, carcass composition and mineral excretion levels of broilers. In a completely randomised experimental design, 900 one-day-old male Ross-308 broilers were assigned to 5 treatments, with 6 replicates of 30 birds each. The control group (CITE) was fed with a basal diet containing regular inclusion levels of inorganic trace elements. Treatment groups were supplied with reduced levels (30% and 50% of the regular level) of inorganic (ITE) or organic trace elements (OTE), respectively. Groups 50% ITE, 30% OTE and 50% OTE diets had equivalent average daily gain (ADG), average daily feed intake (ADFI), feed to gain ratio (F/G ratio) and mortality rate compared with group CITE in any phase. However, compared with group CITE chicks in group 30% ITE have lower ADG and ADFI and higher F/G ratio. The carcass yields were not affected by dietary treatments. Compared with group CITE, in groups 30% ITE, 50% ITE, 30% OTE and 50% OTE the shear force values of the breast muscle were only 71.8%, 83.4%, 63.5% and 59.4% (p < 0.05), respectively. Birds received diets containing reduced levels of trace elements had diminished excretions of Mn and Zn throughout the entire period (p < 0.01). In conclusion, the reduced supplementation of trace elements had no or slightly negative impact on growth performance, carcass yield and meat quality, but decreased faecal mineral excretion. Moreover, the trace element supply as OTE played a limited role on performance and excretion and was only partly beneficial for animal performance in case the trace element supply was reduced to 30%.

Expression characteristics of the yes-associated protein in breast cancer: A meta-analysis.

BACKGROUND: The yes-associated protein (YAP) gene plays an important role in many malignant tumors, but its clinical significance in breast cancer remains unclear. This study aimed to explore the significance of YAP expression in breast cancer using meta-analysis. METHODS: Seven databases will be searched to collect the case-control studies published on the association between YAP expression and clinical pathogenic features in breast cancer until December 2021: PubMed, EMBASE, Web of Science, China National Knowledge Infrastructure, Chinese Scientific Journal Database, Wan Fang Database, and the Chinese Biomedical Literature Database. To perform meta-analysis, STATA 14.0 and RevMan5 software were used with odds ratio (OR) and 95% confidence interval (95% CI) as the effect index, and publication bias and sensitivity analysis were subsequently tested. RESULTS: Form a total of 10 articles used in this study, 8 studies consisted of nontriple negative breast cancer (non-TNBC) and the other 2 of TNBC. Meta-analysis indicated a positive expression rate of YAP in non-TNBC tissues that was lower than in normal breast tissue (OR = 0.15, 95% CI = 0.10-0.21, P < .001). In contrast, the positive rate of YAP expression in TNBC was significantly higher than that in normal breast tissue (OR = 18.23, 95% CI = 8.20-40.52, P < .001). Furthermore, the positive expression rate was higher in the patients with lymph node metastasis, higher tumor node metastasis stage and histologic grade, and larger diameter in TNBC. However, there was no statistical difference in the positive expression rate of YAP between non-TNBC patients and lymph node metastasis, tumor node metastasis stage, histologic grade, and tumor size. CONCLUSIONS: YAP may participate in the occurrence and development of non-TNBC as a tumor suppressor gene; however, it may also be a carcinogenic factor in TNBC and may be a potential therapeutic target for TNBC.

Xylan alleviates dietary fiber deprivation-induced dysbiosis by selectively promoting Bifidobacterium pseudocatenulatum in pigs.

BACKGROUND: Low dietary fiber intake has been shown to disturb the gut microbiome community, damage the mucus barrier, and promote pathogen susceptibility. However, little is known about the temporal response of the gut microbiome to dietary fiber deprivation and the recovery induced by dietary fiber inclusion in pigs. OBJECTIVE: In the present study, temporal responses of ileal and fecal microbiota to dietary fiber deprivation were profiled using an ileum cannulated growing pig model. In addition, the potential of dietary-resistant starch, ß-glucan, and xylan to alleviate gut dysbiosis throughout the gastrointestinal tract, as well as its possible mechanisms were investigated. METHODS: Six cannulated growing pigs were fed a fiber deprivation diet for 35 days. Ileal digesta and feces were collected at days 0, 7, 21, and 35 for 16S rRNA sequencing and short-chain fatty acid (SCFA) determination. Another twenty-four healthy growing pigs were assigned to one of four dietary treatments including (1) fiber-free diet, (2) resistant starch diet, (3) ß-glucan diet, and (4) xylan diet. These twenty-four pigs were fed a corresponding diet for 35 days and slaughtered. Gut microbiome and SCFA concentration were profiled along the gastrointestinal tract. RESULTS: Dietary fiber deprivation-induced consistent microbiota extinction, mainly Bifidobacterium and Lactobacillus, and decreased SCFA concentrations in both ileum and feces. The community structure partially recovered at day 35 compared with baseline while SCFA concentrations remained low. Xylan supplementation alleviated gut dysbiosis by selectively promoting Bifidobacterium pseudocatenulatum within the large intestine. SCFA concentration increased significantly after xylan supplementation and exhibited a positive association with B. pseudocatenulatum abundance. An elevated abundance of xylan degradation-related enzyme genes was also observed in the gut microbiome after xylan supplementation. In vitro growth assay further verified the xylan utilization capacity of B. pseudocatenulatum. CONCLUSIONS: Dietary fiber deprivation could induce probiotic extinction and loss of the SCFA production while potential pathogen was promoted. Xylan intervention could partially restore dietary fiber deprivation-induced gut dysbiosis through selectively promoting B. pseudocatenulatum and therefore normalizing the gut environment. These findings collectively provide evidence that dietary fiber-driven microbiota metabolism bridges the interplay between microbiome and gut health. Video abstract.

Effects of phytonutrients on growth performance, antioxidative status, and energy utilization of broilers fed low energy diets.

Two experiments were conducted to investigate the effects of phytonutrients (PN) on growth performance, antioxidant status, intestinal morphology, and nutrient utilization of birds fed low energy diets. In Exp. 1, a total of 1,440 one-day-old Ross 308 male broiler chickens were randomly divided into 3 treatment groups, with 16 replicates per treatment (48 pens; 30 birds per pen). Birds in treatment 1 were fed diets with normal energy content (NE). Birds in treatment 2 were fed NE diet but with 60 kcal removed (LE). Birds in treatment 3 were assigned to LE diet supplemented with PN (LE + PN). Results indicated that LE diet increased feed conversion ratio (FCR) compared with NE from d 1 to 38, while LE + PN diet prevented this response (P = 0.02). At d 26, birds in the LE + PN group had the highest ileal and jejunal villus height to crypt depth (VH:CD) ratio. At d 39, PN supplementation improved ileal and jejunal VH:CD ratio, compared with LE group. Moreover, birds fed PN diets received a better economic profit. In Exp. 2, 360 one-day-old Ross 308 male broiler chickens were used in a metabolism study. The treatments used in Exp. 2 were the same as those in Exp.1, with 4 replicates (pens) and 30 birds in each replicate. Dietary apparent metabolism energy (AME), energy and protein digestibility were determined between 21 and 28 d of age. Results showed that chickens fed LE + PN diet tended to have greater AME (P = 0.02) and nitrogen-corrected apparent metabolism energy (AMEn) (P = 0.03) than birds fed LE diets. It was concluded that LE + PN showed a potential advantage to improve feed conversion and gut health of broilers, as well as economic profits.

Gene expression alterations of human liver cancer cells following borax exposure.

Borax is a boron compound that is becoming widely recognized for its biological effects, including lipid peroxidation, cytotoxicity, genotoxicity, antioxidant activity and potential therapeutic benefits. However, it remains unknown whether exposure of human liver cancer (HepG2) cells to borax affects the gene expression of these cells. HepG2 cells were treated with 4 mM borax for either 2 or 24 h. Gene expression analysis was performed using Affymetrix GeneChip Human Gene 2.0 ST Arrays, which was followed by gene ontology analysis and pathway analysis. The clustering result was validated using reverse transcription­quantitative polymerase chain reaction. A cell proliferation assay was performed using Celigo Image Cytometer Instrumentation. Following this, 2­ or 24­h exposure to borax significantly altered the expression level of a number of genes in HepG2 cells, specifically 530 genes (384 upregulated and 146 downregulated) or 1,763 genes (1,044 upregulated and 719 downregulated) compared with the control group, respectively (≥2­fold; P<0.05). Twenty downregulated genes were abundantly expressed in HepG2 cells under normal conditions. Furthermore, the growth of HepG2 cells was inhibited through the downregulation of PRUNE1, NBPF1, PPcaspase­1, UPF2 and MBTPS1 (≥1.5­fold, P<0.05). The dysregulated genes potentially serve important roles in various biological processes, including the inflammation response, stress response, cellular growth, proliferation, apoptosis and tumorigenesis/oncolysis.

Surplus dietary isoleucine intake enhanced monounsaturated fatty acid synthesis and fat accumulation in skeletal muscle of finishing pigs.

BACKGROUND: Isoleucine (Ile) has been implicated in the regulation of energy homeostasis and adipogenesis. However, the impact of surplus dietary Ile intake on muscle lipogenesis remains unknown. The present study aimed to investigate the impact of dietary supplementation of extra-Ile on lipogenesis, fatty acid profile and lipid accumulation in skeletal muscle in finishing pigs. METHODS: Forty-eight barrows with initial body weight of 77.0 ± 0.1 kg were allotted to one of two groups and fed diets containing 0.39%, 0.53% standardized ileal digestible (SID) Ile with six replicates per treatment and four pigs per replicate for 30 d. RESULTS: Dietary Ile intake significantly improved the intramuscular fat (IMF) content and monounsaturated fatty acid (MUFA) concentration in the skeletal muscle (P < 0.05), and decreased the drip loss and shear force (P < 0.05) without influencing the growth performance of pigs (P > 0.05). Moreover, the phosphorylation of adenosine monophosphate activated protein kinase α (AMPKα) and acetyl coenzyme A carboxylase (ACC) proteins that monitor lipid metabolism were decreased in skeletal muscle of pigs offered extra-Ile diet (P < 0.05). The mRNA expression of adipose-specific genes adipocyte determination and differentiation factor 1 (ADD1), fatty acid synthase (FAS), and stearoyl-CoA desaturase (SCD) were upregulated and the activity of SCD was increased as well (P < 0.05). CONCLUSIONS: Surplus dietary Ile intake could increase IMF accumulation and MUFA synthesis in skeletal muscle through depressing the phosphorylation of AMPKα-ACC and stimulating the expression and activity of SCD, and increasing the capability of lipogenesis in skeletal muscle.

Enzyme-free ultrasensitive fluorescence detection of epithelial cell adhesion molecules based on a toehold-aided DNA recycling amplification strategy.

Epithelial cell adhesion molecules (EpCAMs) play a significant role in tumorigenesis and tumor development. EpCAMs are considered to be tumor signaling molecules for cancer diagnosis, prognosis and therapy. Herein, an enzyme-free and highly sensitive fluorescent biosensor, with a combined aptamer-based EpCAM recognition and toehold-aided DNA recycling amplification strategy, was developed for sensitive and specific fluorescence detection of EpCAMs. Due to highly specific binding between EpCAMs and corresponding aptamers, strand a, which is released from the complex of aptamer/strand a in the presence of EpCAMs which is bound to the corresponding aptamer, triggered the toehold-mediated strand displacement process. An amplified fluorescent signal was achieved by recycling strand a for ultrasensitive EpCAM detection with a detection limit as low as 0.1 ng mL-1, which was comparable or superior to that of reported immunoassays and biosensor strategies. In addition, high selectivity towards EpCAMs was exhibited when other proteins were selected as control proteins. Finally, this strategy was successfully used for the ultrasensitive fluorescence detection of EpCAMs in human serum samples with satisfactory results. Importantly, the present strategy may be also expanded for the detection of other targets using the corresponding aptamers.

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).

Impact of chemokine receptor CXCR3 on tumor-infiltrating lymphocyte recruitment associated with favorable prognosis in advanced gastric cancer.

Chemokine receptor CXCR3 has been proved to play an important role in tumorigenesis and tumor progression in many malignancies, but its precise efficacy on gastric cancer (GC) has not been evaluated yet. The present study was aimed to explore the correlation of chemokine receptor CXCR3 with tumor-infiltrating lymphocytes (TILs) and prognosis in advanced gastric cancer (GC). Expression of CXCR3 and CD4+, CD8+ TILs was conducted in 192 advanced GC specimens and 48 corresponding paracancerous tissues by immunohistochemical (IHC) analysis. CXCR3 expression in GC tissues was significantly higher than that in paracancerous tissues (P<0.001) and CD8+, CD4+ TILs infiltration increased with high CXCR3 expression (P=0.032 and P<0.001, respectively). Our study showed significantly lower CXCR3 expression in patients with greater tumor invasion depth and lymph node metastasis compared with patients with lesser tumor invasion depth and without lymph node metastasis (P=0.002 and P=0.001, respectively). Univariate analysis indicated that patients with high CXCR3 expression and high CD8+ TILs infiltration had longer overall survival (OS) (log-rank test, P<0.001 and P=0.002, respectively). Univariate and multivariate analyses indicated that CXCR3 expression was an independent prognostic factor for OS (P=0.002). The present study suggested that CXCR3 expression was upregulated in advanced GC and was associated with increased CD4+, CD8+ TILs infiltration and improved OS. Therefore, CXCR3 overexpression is implicated as a favorable prognostic biomarker in human advanced GC.

Determination of energy and amino acid digestibility in growing pigs fed corn distillers' dried grains with solubles containing different lipid levels.

Two experiments were conducted to estimate the digestibility of energy, nitrogen and amino acids (AA) in growing pigs fed diets containing one of five corn distillers' dried grains with solubles (DDGS), including three normal oil DDGS (NO-DDGS) and two low oil DDGS (LO-DDGS) samples. Exp. 1 was conducted to determine the digestible energy (DE) and metabolisable energy (ME) content. Six growing barrows (initial body weight [BW]: 35.1 +/- 2.2 kg) were allotted to a 6 x 6 Latin square design, with six periods and six diets. One diet was a corn soybean meal basal diet and the other five diets were based on corn, soybean meal and 28.8% DDGS. The average DE and ME values for the three NO-DDGS samples were 16.0 and 14.9 MJ/kg dry matter (DM). These values were 9 and 13% greater than the LO-DDGS values of 14.7 and 13.2 MJ/kg DM respectively. Exp. 2 was conducted to determine and compare apparent (AID) and standardised (SID) ileal digestibility for crude protein and AA in the five DDGS samples. Six growing barrows (initial BW, 32.2 +/- 1.9 kg) fitted with a simple T-cannula were allotted to a 6 x 6 Latin square design with six periods and six diets. Five of the diets were based on the five DDGS samples, and the remaining one diet was nitrogen-free diet based on cornstarch and sucrose. Titanium dioxide (0.1%) was used as inert marker. The results of the experiment showed the largest variation among the different samples in AID and SID for lysine (from 41.8 to 65.8% and 53.8 to 73.9% respectively) and threonine (from 54.3 to 73.8% and 65.2 to 79.5% respectively). Also, among the indispensable AA, the SID values for arginine, histidine, threonine and tryptophan observed in LO-DDGS were not different from the values derived from NO-DDGS. In conclusion, LO-DDGS may have decreased energy compared with NO-DDGS because of its lower fat content. However, oil removal during the production of DDGS may not affect amino acid digestibility.