[Association between HBV viral load and severity of liver inflammation in patients with chronic hepatitis B virus infection].

Objective: To explore the relationship and dynamic changes between virological markers and hepatic pathological damage due to host anti-hepatitis B virus (HBV) immunity in the natural course of disease in chronic HBV infected patients. Methods: Two hundred and thirty-eight adult chronic HBV-infected patients who underwent liver biopsy from January 2016 to June 2022 in Taizhou Hospital, Zhejiang Province, were retrospectively selected. General clinical data such as age, gender, platelets, ALT, AST, albumin, HBV DNA, qHBsAg, HBeAg, and liver pathology diagnostic indexes such as the grade of liver necroinflammation and liver fibrotic stages of the patients were collected. The patients were grouped according to HBeAg status, and subgrouped according to different grades of liver necroinflammation and different HBV DNA loads. Statistical analyses were performed to compare the differences in HBV virologic marker levels between the groups, and the correlation between them and the indicators of hepatic inflammatory injury, such as ALT,AST, and the grade of liver necroinflammation in the patients. Results: The levels of HBV virological markers in HBeAg-positive patients with moderate or higher liver necroinflammatory grade (G≥2) were significantly lower than those with mild (no) liver necroinflammatory grade (G < 2) (P < 0.01); whereas the opposite trend was observed in HBeAg-negative patients, with the levels of HBV DNA, and qHBsAg in the G≥2 subgroup being significantly higher than those in the G < 2 subgroup (P < 0.01). Correspondingly, HBV DNA level and qHBsAg showed weak to moderately strong negative correlation with liver necroinflammatory grade and AST which was an indicator of hepatic inflammatory injury in HBeAg-positive patients (P < 0.05); whereas in HBeAg-negative patients, they showed weak to moderately strong positive correlation with hepatic inflammatory activity and ALT, AST (P < 0.001), in which qHBsAg showed only a weak positive correlation with patients' liver necroinflammatory grade (P = 0.003). Further subgroup analyses of HBeAg-positive patients according to whether the HBV DNA level was > 2×10(6) IU/ml showed weak to moderate negative correlations between HBV virological markers and liver necroinflammatory grade as well as ALT and AST in the subgroup of patients with HBV DNA > 2×10(6) IU/ml (P < 0.05); however, the negative correlation disappeared in patients who were still HBeAg positive and had HBV DNA ≤ 2×10(6) IU/ml. Moreover, HBV DNA and ALT, HBeAg and AST showed moderate positive correlation (P < 0.05). Conclusion: We speculate that the activation of host anti-HBV immunity can efficiently inhibit HBV replication by targeting the infected hepatocytes, but only in the early phase of disease progression in HBeAg positive patients with HBV DNA high (> 2×10(6) IU/ml).

Dynamics and bifurcations in multistable 3-cell neural networks.

We disclose the generality of the intrinsic mechanisms underlying multistability in reciprocally inhibitory 3-cell circuits composed of simplified, low-dimensional models of oscillatory neurons, as opposed to those of a detailed Hodgkin-Huxley type [Wojcik et al., PLoS One 9, e92918 (2014)]. The computational reduction to return maps for the phase-lags between neurons reveals a rich multiplicity of rhythmic patterns in such circuits. We perform a detailed bifurcation analysis to show how such rhythms can emerge, disappear, and gain or lose stability, as the parameters of the individual cells and the synapses are varied.

Glucose and lipid metabolism of broiler chickens fed diets with graded levels of corn resistant starch.

1. The aim of this study was to investigate the effects of graded levels of dietary corn resistant starch (RS) on glucose and lipid metabolism of broilers. 2. A total of 320 male broiler chicks (Arbor Acres, one-day-old) were randomly allocated to five dietary treatments, including a corn-soybean control diet, a corn-soybean based diet containing 20% corn starch, and three diets containing 4%, 8% and 12% RS by replacing corn starch with 6.67%, 13.33% and 20% Hi-Maize® 260 (identified as control, RS1, RS2, RS3 and RS4, respectively). Each treatment contained eight replicates with eight birds, and the experiment lasted 42 days. 3. Birds fed RS diets showed lower (P < 0.05) concentrations of serum low-density lipoprotein cholesterol and non-esterified fatty acid (NEFA) at d 21 and 42 of age, compared to the control. Lower (P < 0.05) hepatic apolipoprotein B concentration and citrate synthase (CS) activity, as well as a higher (P < 0.05) glycogen synthase (GS) concentration were observed in birds fed RS diets than those in the control group at d 21 of age. Consuming RS diets linearly increased (P < 0.01) serum glucose concentration, and linearly decreased (P < 0.01) NEFA concentrations in broilers at d 21 and 42 of age. Liver GS concentration and activities of hexokinase, pyruvate and CS were linearly increased (P < 0.01) in broilers at d 21 of age, but were linearly decreased (P < 0.05) in birds at d 42 of age in response to the increase of dietary RS levels. Feeding RS diets linearly decreased (P < 0.05) mRNA expressions of PC, PPARα and CPT-1 at d 21 of age and the mRNA expressions of SREBP-1 c, ChREBP, ACC and FAS at d 42 of age, and linearly increased (P < 0.05) the mRNA expressions of PEPCK, PC, LKB1, AMPKα1, PPARα, CPT-1 and L-FABP at d 42 of age. 4. Feeding broilers with diets containing higher concentration of RS promoted hepatic lipolysis and gluconeogenesis through activated AMPK signalling pathway and accelerated whole-body energy expenditures in the grower phase.

Growth performance, carcass traits and digestive function of broiler chickens fed diets with graded levels of corn resistant starch.

1. This study was conducted to assess the effects of graded levels of dietary corn resistant starch (RS) on growth performance, carcass traits, nutrient retention, digestive organ index, intestinal morphology, digestive enzyme activities, and mRNA expression of certain nutrient transporters in broiler chickens.2. A total of 320, 1-d-old Arbor Acres broiler chickens were randomly assigned to five dietary treatments, with eight replicates of eight birds in each. These treatments included one corn-soybean control diet, a corn-soybean based diet containing 20% corn starch, and three diets supplemented with 4%, 8% and 12% RS by replacing corn starch with 6.67%, 13.33% and 20% of Hi-Maize 260® (identified as control, RS1, RS2, RS3 and RS4, respectively). The feeding period lasted 42 days.3. Performance parameters including feed consumption, feed conversion, body weight gain and percentage of abdominal fat at d 42 of age, nutrient retention (including dry matter, fat, total starch and nitrogen free extract), and apparent metabolisable energy was measured from d 18 to 20 and d 39 to 41 and showed negative linear responses to increasing dietary RS level (P < 0.05). Birds fed the RS3 and RS4 diets showed higher relative weight of duodenum, jejunum and ileum, as well as lower villus height and villus height/crypt depth compared to the control (P < 0.05). The activity of pancreatic trypsin of birds at d 21 and 42 of age decreased linearly in response to the increase of dietary RS level (P < 0.01). There were linear changes in up-regulated mRNA expression of SGLT-1 and down-regulated mRNA expression of GLUT-2 with increasing proportion of RS at d 21 and 42 of age (P < 0.05), respectively.4. It was concluded that feeding broilers with diets containing higher concentrations of RS impaired the development of small intestine, which resulted in lower apparent total tract retention of nutrients and poorer body weight gain, feed efficiency and carcass traits of broiler chickens.

Dietary taurine supplementation enhances antioxidative capacity and improves breast meat quality of broiler chickens.

1. The aim of this study was to determine the effect of dietary taurine supplementation on growth performance, meat quality and antioxidant responses in breast muscle of broiler chickens.2. A total of 72 Arbor Acres broiler chickens (28-day-old) with similar body weight were randomly allocated into two groups, and fed either 0 g/kg (control; C) or 5 g/kg taurine-supplemented diets (TS) for 14 days.3. The results showed that TS had no effect on growth performance or chemical composition of breast muscle in broilers. The drip and cooking losses were significantly decreased (P < 0.05), and the pH24h of breast muscle were increased (P < 0.05) in the TS group. Meanwhile, broilers in the TS group exhibited significantly higher (P < 0.05) scavenging activities of superoxide and 2,2-diphenyl-1-picrylhydrazyl radicals, and lower (P < 0.05) contents of carbonyl, malondialdehyde, and 4-hydroxynonenal. TS increased (P < 0.05) total antioxidant capacity and superoxide dismutase activities. Moreover, TS significantly upregulated (P < 0.05) the mRNA expressions of nuclear factor erythroid 2-related factor 2 (Nrf2), heme oxygenase 1, NAD(P)H quinone dehydrogenase 1, superoxide dismutase, catalase, and glutathione peroxidase.4. These findings suggested that TS enhanced antioxidative capacity and improved breast meat quality of broilers via activating the Nrf2 pathway.

Spatial association between outdoor air pollution and lung cancer incidence in China.

BACKGROUND: Lung cancer is the most common cancer in China. Previous studies have indicated that lung cancer incidence exhibits remarkable spatial heterogeneity, and lung cancer is related to outdoor air pollution. However, the non-linear spatial association between outdoor air pollution and lung cancer incidence in China remains unclear. METHODS: In this study, the relationships between the lung cancer incidence of males and females from 207 counties in China in 2013 with annual concentrations of PM2.5, PM10, SO2, NO2, CO and O3 were analysed. GeoDetector q statistic was used for examining the non-linear spatial association between outdoor air pollution and incidence of lung cancer. RESULTS: An apparent spatial and population gender heterogeneity was found in the spatial association between outdoor air pollution and lung cancer incidence. Among the six selected pollutants, SO2 has the greatest influence on lung cancer (q = 0.154 in females) in north China. In the south, each selected pollutant has a significant impact on males or females, and the mean q value in the south is 0.181, which is bigger than that in the north (q = 0.154). In addition, the pollutants have evident non-linear interaction effects on lung cancer. In north China, the interaction between SO2 and PM2.5 is the dominant interaction, with q values of 0.207 in males and 0.334 in females. In the south, the dominant interactive factors are between SO2 and O3 in males and between SO2 and CO in females, with q values of 0.45, 0.232 respectively. Smoking is a substantial contributor to lung cancer among men, either in South or North China, with q value of 0.143 and 0.129 respectively, and the interaction between smoking and air pollutants increases this risk. CONCLUSIONS: This study implies that the influence of SO2 and PM2.5 on lung cancer should be focused on in north China, and in the south, the impact of O3 and CO as well as their interaction with SO2 need to be paid more attention. Smoking, particularly in men, remains a significant risk factor for lung cancer in both North and South China.

Identifying the location of epidermal growth factor-responsive element involved in the regulation of type IIb sodium-phosphate cotransporter expression in porcine intestinal epithelial cells.

Phosphate is an important mineral nutrient for both human and animals in growth and physiological functions; thus, much effort in the past has been made to clarify the mechanisms governing its absorption. Previous studies have found that epidermal growth factor (EGF) inhibits phosphate absorption in human intestinal cells via modulating the interaction of transcriptional factor c-myb with sodium-phosphate cotransporter (NaPi-IIb) gene promoter. This finding provoked our interest in determining the effect of EGF on NaPi-IIb gene expression in intestinal cells of pigs and the location of EGF-responsive element in the gene promoter. Using quantitative PCR, it was observed that EGF significantly reduced NaPi-IIb gene expression in porcine intestinal epithelial IPEC-J2 cells. Transfection with a series of constructs that contain different lengths of the 5'-flanking promoter region of the NaPi-IIb gene manifested that EGF-responsive element is located in the -1200 to -800 region. Further, c-myb was extracted from the cell nucleus of IPEC cells that were exposed to EGF or not via immunoprecipitation. The electrophoretic mobility shift assay showed a specific binding of transcription factor c-myb to labelled probes encompassing DNA sequence from -1092 to -1085 (-TCCAGTTG-). This protein-DNA complex was decreased with cells exposed to EGF and abrogated when c-myb was pre-incubated with excessive unlabelled competitive probes. Results from mutagenesis studies demonstrated that the c-myb-binding site is the EGF-responsive element involved in the regulation of NaPi-IIb expression. Identifying the location of EGF-responsive element contributes to understanding mechanisms underlying EGF down-regulated NaPi-IIb gene expression and provides a foundation for further investigating EGF-regulatory functions in phosphate absorption in pig intestine.

Effects of Water-misting Sprays with Forced Ventilation after Transport during Summer on Meat Quality, Stress Parameters, Glycolytic Potential and Microstructures of Muscle in Broilers.

Effects of water-misting sprays with forced ventilation after transport during summer on meat quality, stress parameters, glycolytic potential and microstructures of muscle in broilers were investigated. A total of 105 mixed-sex Arbor Acres broilers were divided into three treatment groups: i) 45-min transport without rest (T group), ii) 45-min transport with 1-h rest (TR group), iii) 45-min transport with 15-min water-misting sprays with forced ventilation and 45-min rest (TWFR group). The results showed the TWFR group significantly increased (p<0.05) initial muscle pH (pHi) and ultimate pH (pHu) and significantly reduced L* (p<0.05), drip loss, cook loss, creatine kinase, lactate dehydrogenase activity, plasma glucose content, lactate and glycolytic potential when compared with other groups. Microstructure of the muscle from TWFR group broilers under light microscopy showed smaller intercellular spaces among muscle fibers and bundles compared with T group. In conclusion this study indicated water-misting sprays with forced ventilation after transport could relieve the stress caused by transport under high temperature, which was favorable for the broilers' welfare. Furthermore, water-misting sprays with forced ventilation after transport slowed down the postmortem glycolysis rate and inhibited the occurrence of PSE-like meat in broilers. Although rest after transport could also improve the meat quality, the effect was not as significant as water-misting sprays with forced ventilation after transport.

Evolution of CD4⁺CD25(hi) T cell subsets in Aspergillus-infected liver transplantation recipients reduces the incidence of transplantation rejection via upregulating the production of anti-inflammatory cytokines.

Infection is the leading risk factor of liver transplantation-related death. Aspergillosis is a life-threatening complication in immune-compromised patients, and is the cause of approximately 2/3 of deaths in liver transplant recipients. In our previous studies, we found a regulatory T cell (Treg) population that showed significantly increased immune tolerance in Aspergillus-infected liver transplant recipients. Furthermore, interleukin (IL)-17 production was also increased, and an IL-17-producing Treg cell subset was identified in these patients. Functional studies of the role of these IL-17-producing Treg cells in the induction of immune tolerance are needed to help reduce the death rate of liver transplantation recipients. This study included 75 liver transplant recipients with and without histologically confirmed aspergillosis after liver transplantation. The percentage of T cell population subsets producing cytokines was detected by fluorescence-activated cell sorting and enzyme-linked immunosorbent assay in peripheral blood. Complements in blood serum were also examined. The risk of acute rejection was lower in Aspergillus-infected liver transplant recipients compared to the non-Aspergillus-infected group; the CD4(+)CD25(hi) T cell population in peripheral blood was higher and the CD4(+)CD45RA-CD45RO(+) T cell population was lower. There was no significant difference between the CD4(+)CD25(lo)CD45RA(+) and CD4(+)CD25(lo)CD45RA- T cell populations. Moreover, IL-6 decreased and IL-4 increased in the blood serum of Aspergillus-infected liver transplant recipients. Together, these results indicate that the incidence of graft rejection in liver transplantation recipients with Aspergillus infections was lower than that of the non-infected group, and suggests a mechanism for this effect.

Expression and clinical significance of miR-122 and miR-29 in hepatitis B virus-related liver disease.

MicroRNA molecules have been increasingly regarded as a diagnostic and prognostic marker of certain diseases. The aim of this study was to investigate the expression and clinical significance of miR-122 and miR-29 in liver disease related to hepatitis B virus infection. The serum levels of miR-122 and miR-29 in 20 patients with hepatocellular carcinoma (HCC), 20 patients with liver cirrhosis (LC), 29 patients with chronic hepatitis B (CHB), 20 cases of hepatitis B virus carriers (ASC), and 20 healthy controls (HC) were determined by a fluorescence real-time quantitative PCR method and then evaluated by clinical correlation analysis. Compared with the serum levels of miR-122 in the HC, LC, and ASC groups, those in patients with HCC and CHB were significantly increased. The serum levels of miR-29 in LC patients were lower than those in the healthy controls (P < 0.01). A positive correlation was observed between the expression of miR-122 and miR-29, and HBV DNA in patients with CHB. A negative correlation was found between miR-29 and α-fetoprotein in patients with HCC. The elevation in miR-122 was correlated with liver damage in CHB patients and with the pathogenesis of liver cancer in HCC patients. The decrease in miR-29 expression was related to the incidence of liver fibrosis. The detection of miR-122 and miR-29 may be useful in evaluating the inflammatory liver injury and fibrosis associated with chronic HBV infection.

DNA methylation promotes the expression of PPARγ transcript 1 at least in part by preventing NRF1 binding to the promoter P1 of chicken PPARγ gene.

Peroxisome proliferator-activated receptor gamma (PPARγ) is a master regulator of adipogenesis. Our previous study revealed that chicken PPARγ has 3 alternative promoters named as P1, P2, and P3, and the DNA methylation of promoter P3 was negatively associated with PPARγ mRNA expression in abdominal adipose tissue (AAT). However, the methylation status of promoters P1 and P2 is unclear. Here we assessed promoter P1 methylation status in AAT of Northeast Agricultural University broiler lines divergently selected for abdominal fat content (NEAUHLF). The results showed that promoter P1 methylation differed in AAT between the lean and fat lines of NEAUHLF at 7 wk of age (p < 0.05), and AAT expression of PPARγ transcript 1 (PPARγ1), which was derived from the promoter P1, was greatly higher in fat line than in lean line at 2 and 7 wk of age. The results of the correlation analysis showed that P1 methylation was positively correlated with PPARγ1 expression at 7 wk of age (Pearson's r = 0.356, p = 0.0242), suggesting P1 methylation promotes PPARγ1 expression. To explore the underlying molecular mechanism of P1 methylation on PPARγ1 expression, bioinformatics analysis, dual-luciferase reporter assay, pyrosequencing, and electrophoresis mobility shift assay (EMSA) were performed. The results showed that transcription factor NRF1 repressed the promoter activity of the unmethylated P1, but not the methylated P1. Of all the 4 CpGs (CpG48, CpG49, CpG50, and CpG51), which reside within or nearby the NRF1 binding sites of the P1, only CpG49 methylation in AAT was remarkably higher in the fat line than in lean line at 7 wk of age (3.18 to 0.57, p < 0.05), and CpG49 methylation was positively correlated with PPARγ1 expression (Pearson's r = 0.3716, p = 0.0432). Furthermore, EMSA showed that CpG49 methylation reduced the binding of NRF1 to the P1. Taken together, our findings illustrate that P1 methylation promotes PPARγ1 expression at least in part by preventing NRF1 from binding to the promoter P1.

[Efficacy and safety analysis of P-GemDOx regimen and stratified prognosis in patients with early extranodal NK/T cell lymphoma].

Objective: To assess the efficacy, safety, and related prognostic factors associated with the P-GemDOx regimen as a first-line treatment for patients with early-stage extranodal natural killer (NK) /T cell lymphoma (ENKTL) . Methods: A retrospective analysis was performed on sixty early-stage ENKTL patients treated with the P-GemDOx regimen who were admitted to the First Affiliated Hospital of Nanjing Medical University between August 2015 and May 2021. The Chi-square test or Fisher's exact test was used to compare group differences, and the Log-rank test was used to compare the differences in survival. Survival outcomes and prognostic factors were examined. Results: After completing 4 to 6 cycles of P-GemDOx chemotherapy, the overall response rate (ORR) was 88.3%, with forty-six patients (76.7% ) achieving complete response (CR). The 4-year progression-free survival (PFS) and overall survival (OS) rates were (66.3±7.1) % and (79.5±6.0) %, respectively. According to the PINK/PINK-E model, there was no significant difference in survival outcomes among risk groups. 23.3% of patients experienced progression of disease within 24 months (POD<24). OS estimates differed significantly (P<0.001) between the POD<24 group (n=14) and the POD≥24 group (n=46). Analysis showed that SUVmax > 12.8 at diagnosis, non-single nasal cavity infiltration, and response less than CR after 4-6 cycles all had a significant association with POD24. We used these data as the basis for predicting POD<24 international prognostic index (POD24-IPI). Patients were stratified into low-risk (no risk factors), intermediate-risk (one risk factor), or high risk (two or three risk factors). These groups were associated with 4-year OS rate of 100%, (85.6±9.7) %, and (65.0±10.2) %, respectively (P=0.014). The P-GemDOx regimen was well tolerated, with hematological toxicity being the main side effect. Conclusion: This study demonstrated that the P-GemDOx regimen is effective and safe in the first-line treatment of early-stage ENKTL, and POD24-IPI is a promising prognostic model.

Prevalence of multidrug-resistant gram-negative bacilli producing extended-spectrum ß-lactamases (ESBLs) and ESBL genes in solid organ transplant recipients.

BACKGROUND: Multidrug-resistant (MDR) gram-negative bacilli pose a serious and rapidly emerging threat to recipients of solid organ transplants (SOTs). However, extended-spectrum ß-lactamases (ESBLs), as one of the cardinal mechanisms of resistance to antimicrobial agents in SOT recipients, remain obscure. The aim of this study was to investigate the prevalence of strains producing ESBLs in SOT patients with MDR gram-negative bacilli infections and to identify the ESBL genes carried by them. MATERIALS AND METHODS: Isolates from various clinical specimens of SOT recipients were identified using the VITEK 2 system, and their antibiotic sensitivity was determined by the 2-fold agar dilution method. Isolates suspected of producing ESBL enzymes were tested by an ESBL phenotypic confirmatory method, and the ESBL genotypes were determined by the polymerase chain reaction and sequencing. RESULTS: A total of 80 MDR gram-negative strains were isolated from 350 SOT recipients. Among these strains, 42 (52.5%) had an ESBL-positive phenotype; 33 (42.3%) patients with ESBL-positive infection were found. Molecular analysis showed that most of the isolates harbored blaCTX-M-9 (78.6%), blaTEM (81.0%), and blaSHV (69.0%) genes. All the ESBL-producing strains were susceptible to carbapenems. CONCLUSIONS: MDR gram-negative isolates from SOT recipients are frequently ESBL producers. TEM and CTX-M9 were the predominant ESBL types.

Methylation regulation of liver-specific microRNA-122 expression and its effects on the proliferation and apoptosis of hepatocellular carcinoma cells.

The regulation mechanism and significance of microRNA-122 (miRNA-122) expression are unclear. The aim of this study was to investigate the effects of DNA methylation on liver-specific miRNA-122 expression, cell proliferation, and apoptosis in hepatocellular carcinoma. Methylation of the miRNA-122 promoter region was detected through methylation sequencing. The level of miRNA-122 expression was measured using real-time quantitative polymerase chain reaction. The proliferation and apoptosis of hepatocellular cell lines were detected using flow cytometry and Cell Counting Kit-8 assays. Compared with those in human primary hepatocytes, methylation levels of the miRNA-122 promoter in the Huh7, HepG2, and QSG-7701 cell lines were significantly increased (P = 0.000). Similarly, levels of miRNA-122 expression in these cell lines significantly decreased (P = 0.007). After treatment with 5-aza-2-deoxycytidine, the Huh7 and HepG2 cell lines displayed a significantly lower degree of methylation (P = 0.038 and 0.025), and the levels of miRNA-122 expression were significantly higher (P = 0.008 and 0.003) than those in the blank group. Compared with the blank group, apoptosis of Huh7 and HepG2 cells was significantly increased (P = 0.001 and 0.027). We concluded that the expression of miRNA-122 is regulated by DNA methylation and correlated with apoptosis of liver cancer cells. Methylation regulation of miRNA-122 expression might be involved in the development of hepatocellular carcinoma.

Cadmium toxicity is alleviated by AtLCD and AtDCD in Escherichia coli.

AIMS: Arabidopsis thaliana l- and d-cysteine desulfhydrases (AtLCD and AtDCD) are two important H(2) S-generating enzymes. This study determined the effects of H(2) S derived from AtLCD and AtDCD on cadmium (Cd) toxicity in Escherichia coli. METHODS AND RESULTS: AtLCD and AtDCD were cloned into pET28a vectors and transformed into wild-type E. coli strain BL21(DE3), named BL21(LCD) and BL21(DCD). In the induced BL21(LCD) and BL21(DCD) compared with wild type, significantly higher H(2) S generation rates were observed. Additionally, higher survival rates, reduced contents of malondialdehyde (MDA) and hydrogen peroxide (H(2) O(2)), decreased activities of superoxide dismutase and catalase under 220 µmol l(-1) Cd stress were noted. We obtained similar results in the wild type treated with NaHS, a H(2) S donor. The above changes were substantially counteracted by the mixture of ammonia and pyruvic acid potassium (NH(3) + C(3) H(3) KO(3)), a synthetic inhibitor of H(2) S. CONCLUSIONS: AtLCD and AtDCD catalyse the H(2) S production, generating an ameliorating effect against Cd-induced oxidative stress and resulting in E. coli resistance to Cd toxicity. SIGNIFICANCE AND IMPACT OF THE STUDY: H(2) S as a gasotransmitter is certified to have an ameliorating effect against Cd toxicity, thus providing information for further research regarding the role of H(2) S in regulating resistance to the heavy metal stress in organisms.

Creatine nitrate supplementation strengthens energy status and delays glycolysis of broiler muscle via inhibition of LKB1/AMPK pathway.

This study aimed to evaluate the effects of dietary creatine nitrate (CrN) on growth performance, meat quality, energy status, glycolysis, and related gene expression of liver kinase B1/AMP-activated protein kinase (LKB1/AMPK) pathway in Pectoralis major (PM) muscle of broilers. A total of 240 male Arbor Acres broilers (28-day-old) were randomly allocated to one of 5 dietary treatments: the basal diet (control group), and the basal diets supplemented with 600 mg/kg guanidinoacetic acid (GAA), 300, 600, or 900 mg/kg CrN (identified as GAA600, CrN300, CrN600, or CrN900, respectively). We found that dietary GAA and CrN supplementation for 14 d from d 28 to 42 did not affect broiler growth performance, carcass traits, and textural characteristics of breast muscle. GAA600, CrN600, and CrN900 treatments increased pH24h and decreased drip loss of PM muscle compared with the control (P < 0.05). The PM muscles of CrN600 and CrN900 groups showed higher glycogen concentration and lower lactic acid concentration accompanied by lower activities of phosphofructokinase (PFK), pyruvate kinase (PK), and lactate dehydrogenase (LDH) (P < 0.05). Simultaneously, GAA600 and all CrN treatments increased concentration of muscle creatine, phosphocreatine (PCr) and ATP, and decreased AMP concentration and AMP/ATP ratio (P < 0.05). Meanwhile, the concentrations of muscle creatine, PCr, and ATP were increased linearly, while muscle AMP concentration and AMP/ATP ratio were decreased linearly and quadratic as the dose of CrN increased (P < 0.05). GAA600, CrN600, and CrN900 treatments upregulated mRNA expression of CreaT in PM muscle, and CrN600 and CrN900 treatments downregulated GAMT expression in liver and PM muscle compared with the control or GAA600 groups (P < 0.05). The mRNA expression of muscle LKB1, AMPKα1, and AMPKα2 was downregulated linearly in response to the increasing CrN level (P < 0.05). Overall, CrN showed better efficacy on strengthening muscle energy status and improve meat quality than GAA at the some dose. These results indicate that CrN may be a potential replacement for GAA as a new creatine supplement.

The combined impact of xylo-oligosaccharides and gamma-irradiated astragalus polysaccharides on the immune response, antioxidant capacity, and intestinal microbiota composition of broilers.

The present study investigated the individual and combined effects of xylo-oligosaccharides (XOS) and gamma-irradiated astragalus polysaccharides (IAPS) on the immune response, antioxidant capacity and intestinal microbiota composition of broiler chickens. A total of 240 newly hatched Ross 308 chicks were randomly allocated into 5 dietary treatments including the basal diet (control), or the basal diet supplemented with 50 mg/kg chlortetracycline (CTC), 100 mg/kg XOS (XOS), 600 mg/kg IAPS (IAPS), and 100 mg/kg XOS + 600 mg/kg IAPS (XOS + IAPS) respectively. The results showed that birds in the control group had lower the thymus index and serum lysozyme activity than those in the other 4 groups (P < 0.05). Moreover, there was an interaction between XOS and IAPS treatments on increasing the serum lysozyme activity (P < 0.05). Birds in the CTC and XOS + IAPS groups had lower serum malondialdehyde concentration and higher serum total antioxidant capacity activity and mucosal interleukin 2 mRNA expression of jejunum than those in the control group (P < 0.05). In addition, birds in the control groups had lower duodenal and jejunal IgA-producing cells number than these in other 4 groups (P < 0.05). As compared with the CTC group, dietary individual XOS or IAPS administration increased duodenal IgA-producing cells number (P < 0.05). Meanwhile, there was an interaction between XOS and IAPS treatments on increasing duodenal and jejunal IgA-Producing cells numbers (P < 0.05). Dietary CTC administration increased the proportion of Bacteroides, and decreased the proportion of Negativibacillus (P < 0.05). However, dietary XOS + IAPS administration increased Firmicutes to Bacteroidetes ratio, the proportion of Ruminococcaceae, as well as decreased the proportion of Barnesiella and Negativibacillus (P < 0.05). In conclusion, the XOS and IAPS combination could improve intestinal mucosal immunity and barrier function of broilers by enhancing cytokine gene expression, IgA-producing cell production and modulates cecal microbiota, and the combination effect of XOS and IAPS is better than that of individual effect of CTC, XOS, or IAPS in the current study.

Enhanced transforming growth factor-beta signaling and fibrosis in the pectoralis major muscle of broiler chickens affected by wooden breast myopathy.

Fibrosis has also been recorded as a prominent pathological feature within wooden breast (WB) myopathy of broiler chickens. This study was conducted to evaluate the accumulation of fibril collagen, deposition of the extracellular matrix (ECM) components, and the underlying mechanism mediating the pathogenic fibrotic process in the pectoralis major (PM) muscle of WB-affected birds. Broiler chickens were categorized into the control and WB groups based on the evaluation of myopathic lesions. Results indicated that the total content and area of collagen in cross-sections of the PM muscle, as well as the augmented expression of collagen-I and fibronectin in the ECM, were greatly increased in birds with WB. Wooden breast myopathy upregulated expressions of transforming growth factor-beta (TGF-ß) and the phosphorylation of Smad 2 and 3, thereby activating TGF-ß-mediated Smad signaling pathway, which further enhanced the transcription of profibrotic mediators. In addition, regulators involved in collagen biosynthesis and cross-linking including prolyl 4-hydroxylase, lysyl oxidase, lysyl hydroxylase, and decorin were increased in the WB muscle. Finally, the expressions of both matrix metalloproteinases (MMP) and tissue inhibitor of metalloproteinases (TIMP) were increased in the WB muscle, which might be related with reduced ECM remodeling. Overall, WB myopathy induces severe fibrosis by enhancing ECM deposition and collagen cross-linking in the PM muscle of broiler chickens, possibly via the activation of TGF-ß signaling and the dysregulation of the MMP and TIMP system.

Microbiota populations and short-chain fatty acids production in cecum of immunosuppressed broilers consuming diets containing γ-irradiated Astragalus polysaccharides.

This study was designed to evaluate the effects of γ-irradiated Astragalus polysaccharides (IAPS) on growth performance, cecal microbiota populations, and concentrations of cecal short-chain fatty acids of immunosuppressed broilers. A total of 144 one-day-old broiler chicks were randomly assigned into 3 groups: nontreated group (control), cyclophosphamide (CPM)-treated groups fed either a basal diet or the diets containing 900 mg/kg IAPS, respectively. On day 16, 18, and 20, broilers in the control group were intramuscularly injected with 0.5 mL sterilized saline (0.75%, wt/vol), and those in the CPM and IAPS groups were intramuscularly injected with 0.5 mL CPM (40 mg/kg of BW). The trial lasted 21 d. Compared with the control group, CPM treatment decreased the broiler average daily gain (ADG) and feed intake (P < 0.05) but did not affect the overall microbial diversity and compositions, as well as the concentrations of cecal acetate, propionate, and butyrate in cecum of broilers (P > 0.05). Dietary IAPS supplementation increased broiler ADG, Shannon index, and decreased Simpson index (P < 0.05). Specifically, broilers fed diets containing IAPS showed lower abundances of Faecalibacterium, Bacteroides, and Butyricicoccus and higher proportions of Ruminococcaceae UCG-014, Negativibacillus, Shuttleworthia, Sellimonas, and Mollicutes RF39_norank, respectively (P < 0.05). The IAPS treatment also increased butyrate concentration (P < 0.05) and tended to elevate acetate concentration (P = 0.052) in cecal digesta. The results indicated that IAPS are effective in increasing the cecal beneficial bacteria and short-chain fatty acids production, contributing to improvement in the growth performance of immunosuppressive broilers. These findings may expand our knowledge about the function of modified Astragalus polysaccharides in broiler chickens.