The current study aimed to assess the effects of the addition of dietary fermented feed (FF) on the growth performance, blood parameters, intestinal morphology, and number of cecal microorganisms of Sichuan white geese from 1 to 28 d of age. A total of 240, 1-day-old female geese were randomly divided into five groups with six replicates (eight birds per replicate). Geese in the control group were fed the basal diet (0.0% FF) and the experimental groups were fed the basal diet supplemented with 2.5%, 5.0%, 7.5%, and 10.0% FF, respectively. The 5.0% FF and 7.5% FF groups had greater body weights (BW), average daily feed intakes (ADFI), and average daily gains (ADG) than the control group. The feed/gain ratio (F/G) in the 7.5% FF group was higher than the control group. The concentrations of immunoglobulin A (IGA), superoxide dismutase (SOD), catalase, total antioxidant capacity (T-AOC), and glutathione peroxidase (GPx) in the 5.0% FF group were higher than those in the control group, whereas malondialdehyde (MDA) content in the 5.0% FF group was lower. Moreover, the FF did not affect the plasma biochemical indicators, number of cecal microorganisms, and intestinal morphology of geese in the groups. These findings indicate that 5.0% FF may improve the growth performance (BW, ADG, and ADFI), immunological function, and antioxidant capacity of Sichuan white geese from 1 to 28 days of age.

ABSTRACT Geese (Anser cygnoides) possess stronger ability of roughage digestion and utilization than other poultries, hence, it has become the focus of attention of scientists. Duodenal, jejunum and ileum were mainly participated in food digestion and nutrient absorption, while the cecum was responsible for biological fermentation. Effects on the geese’s cecal microbiota community by feeding with the all-grass diet have been investigated, however, whether it had an influence on the geese’s duodenal microbiota community remains unexplored. To address this problem, geese feeding with the basal diet for 28 days (G1), the basal diet for 28 days and the all-grass diet for the following 14 days (G2), the basal diet for 42 days (G3) were selected, respectively. The duodenal segments of geese were collected and the hypervariable V3-V4 region of the bacterial 16S rRNA gene was sequencing. A total of 4 main phyla and 16 main genera were identified. Moreover, we also successfully identified that two taxa including the Helcococcus and Clostridium could be used as distinguishing biomarkers specific to G2. The functional profiles of the duodenum microbiota were mainly involved in the membrane transport (e.g. ABC transporters), amino acid metabolism, energy metabolism, metabolism of cofactors and vitamins, and cellular processes and signaling pathways in geese feeding with the all-grass diet. In conclusion, the all-grass diet could impact the composition of duodenal microbiota. However, to resolve the underlying mechanism of the fiber digesting and utilization in geese’s gut microbiota, the whole intestinal system needs to be assessed by further studies.

To explore the effects of supplementing a basal diet with green sweet sorghum stalks on the gut microbiota in geese, one hundred and twenty 28-day-old geese were divided into two groups. Group 1 was fed a basal diet and group 2 was fed 92% of the same basal diet with green sweet sorghum stalks ad libitum for 42 days. Three male geese from each group were killed at 70 days old, four gastrointestinal tract segments (duodenum, jejunum, ileum, and caecum) were collected, and the hypervariable V4 region of the bacterial 16S rRNA gene was sequenced. There were obvious changes of microbiome in the caecum compared with the other three intestinal segments after green sweet sorghum stalks were added to the diet. Group 2 had significantly higher alpha diversity in the caecum compared with group 1 (P <0.05). The phyla of bacterial communities in the caecum differed. Group 2 had more abundant Bacteroidetes and Firmicutes, but Proteobacteria were more abundant in group 1. At genus level, Bacteroides was more prevalent in group 2, as were Prevotella and YRC22 (P <0.05). Functional analysis revealed that the carbohydrate metabolism, membrane transport, endocrine system and digestive system metabolic pathways were overrepresented in group 2. In conclusion, caecal microbes might play an important role in digesting green sweet sorghum stalks in geese.

Naringin is a major flavanone derivate that has many important biological functions in animals. However, its effect on pigs is unknown. The aim of this study was to investigate the effect of naringin supplementation on performance, carcass traits, meat quality and oxidative stability in finishing pigs. Ninety-six pigs, with an average initial body weight of 66.2 ± 0.63 kg, were randomly divided into four groups. One group was fed a basal diet without supplementation (control), and the three others were fed diets supplemented with 0.5, 1.0 or 1.5 g naringin /kg DM of feed for 50 days. Each treatment was replicated six times with four pigs per replicate. Feed and water were available ad libitum. The 0.5 g/kg naringin treatment group had an improved loin eye muscle area, reduced serum triglycerides and were leaner compared with the other groups. Pigs in the 1.5 g/kg naringin treatment had higher pH45min values and inosine monophosphate concentrations, and lower MyHC IIb mRNA expression in muscle than the other groups. MyHC IIa mRNA expression was significantly up-regulated in all naringin-supplemented diet groups. Naringin significantly increased superoxide dismutase (SOD) activity and total anti-oxidative capacity in meat, as well as SOD and glutathione peroxidase activity in the liver. These results indicate that the dietary addition of naringin at 0.5 g/kg improved carcass characteristics, while 1.5 g/kg improved the oxidative stability and pork quality in finishing pigs.

ABSTRACT The objective of the study was to investigate the mechanism by which dietary energy concentration regulates laying performance in geese. Eighty 558-day-old female Sichuan White geese were randomly allotted to two dietary treatments, each treatment was fed 1 of 2 experimental diets containing 10.00 (deficient) or 11.80MJ/kg metabolizable energy (sufficient) for 30 days. Laying performance, hormone concentration and gene expressions in hypothalamus-pituitary-gonadal axis were examined in geese. Birds fed the sufficient-energy diet had significantly higher average egg weight, daily laying rate, and lower feed to egg ratio than those fed the deficient-energy (p<0.05). The birds fed sufficient-energy diet had higher concentration of serum insulin like growth factor 1 (IGF-1), gonadotropin-releasing hormone (GnRH), follicle-stimulating hormone (FSH) and estradiol (E2) than those in deficient-energy diet (p<0.05). The mRNA expression levels of GnRH in the hypothalamus, FSH in the pituitary and E2 in the ovary of birds fed sufficient-energy diet were higher than the corresponding counterpart in deficient-energy diet (p<0.05), respectively. In conclusion, the study implied that dietary energy modifies laying possibly through regulating reproductive hormone secretion and gene expression in hypothalamus-pituitary-gonad axis in laying geese.

ABSTRACT This study was carried to express the interferon-induced transmembrane protein 3 (IFITM3) in vitro and examine its function in inhibition of avian reovirus (ARV) replication. The recombinant prokaryotic vector expressing yellow-feathered broiler IFITM3 was successfully constructed, and the recombinant protein was expressed in competent Escherichia coli BL21 cells. New Zealand white rabbits were immunized with the purified recombinant protein to prepare a polyclonal antibody, with a titer of 1:128,000. Immunohistochemistry, reverse transcription-PCR, and real-time fluorescence quantitative PCR showed that IFITM3 was distributed in the yellow-feathered broiler immune organs, and the expression of IFITM3 in bursa of Fabricius was more than in spleen and thymus. It was found that in the thymus, spleen and bursa of Fabricius the mRNA expression levels of IFNα and IFITM3 were significantly induced after ARV infection. And it was also certified in the chicken embryo fibroblasts (CEFs) which infected with ARV. Then the IFNα was added into the cell culture medium before CEFs were infected with ARV. The results indicated that the mRNA of IFITM3 expression was significantly increased and ARV multiplication was significantly inhibited. And when the expression of IFITM3 was knocked down by siRNA-IFITM3, the expression of IFITM3 was significantly reduced, but the ARV multiplication was significantly increased, which indicated that IFITM3 protein could inhibit the ARV replication.

This study aimed to investigate the mechanism of hypoxia-inducible factor-1 alpha (HIF-1α) mediated hypoxia-induced permeability changes in bladder endothelial cells. Models of in vitro hypoxic cell culture of bladder cancer, bladder cancer cells with low HIF-1α expression and HIF-1α RNA interference (RNAi) expression vector were established. Western blot and reverse transcription polymerase chain reaction (RT-PCR) were used to detect the expression of HIF-1α and vascular endothelial growth factor (VEGF) in each group. Bladder cell permeability was determined. Results showed that protein and mRNA expression of HIF-1α and VEGF at 3 and 12 h of hypoxia were significantly higher than normal control (P<0.05), and peaked at 12 h. HIF-1α and VEGF expression in the hypoxic group and hypoxic+3-(5′-hydroxymethyl-2′-furyl)-1-benzyl indazole (YC-1) group were significantly higher than normal control (P<0.05), while expression in the hypoxic+YC-1 group was significantly lower than the hypoxic group (P<0.05). Bladder cell permeability in the hypoxic and hypoxic+YC-1 group were significantly increased compared to normal control (P<0.05), while in the hypoxic+YC-1 group was significantly decreased compared to the hypoxic group (P<0.05). Most of the cells in the stably transfected HIF-1α RNAi expression vector pcDNA6.2-GW/EmGFP-miR-siHIF-1α expressed green fluorescence protein (GFP) under fluorescence microscope. pcDNA6.2-GW/EmGFP-miR-siHIF-1α could significantly inhibit HIF-1α gene expression (P<0.05). HIF-1α and VEGF expression in the hypoxic group and siHIF-1α hypoxic group were significantly higher than normal group (P<0.05), while expression in the siHIF-1α hypoxic group was significantly lower than the hypoxic group (P<0.05). Findings suggest that HIF-1α is an important factor in the increase of bladder cancer cell permeability.

ABSTRACT This experiment was conducted to investigate the effects of Bacillus coagulans on the growth performance and immune functions of the intestinal mucosa of yellow broilers. Three hundred and sixty one-day-old yellow chicks were randomly allocated to four treatments groups with six replicates of 15 chicks each. The broilers were randomly subjected to one of the following treatments for 28 days: control group (group1, fed a basal diet) and three treatments (group 2, 3, 4) fed the basal diet supplemented with 100, 200, or 300 mg/kg Bacillus coagulans , respectively). The results showed that for 28 days, compared with the control diet, the dietary addition of 200 mg/kg Bacillus coagulans significantly decreased the feed/gain ratio (F/G) (p<0.05), improved the thymus index, spleen index and bursa index (p<0.05), increased the villus height to crypt depth ratio (V/C) in the duodenum (p<0.05), increased the number of secretory immunoglobulin (sIgA) positive cells ( p<0.05). The dietary addition of 200 mg/kg Bacillus coagulans promoted a significant increase in Lactobacillus spp. populations and suppressed Escherichia coli replication in cecum, compared with the control (p<0.05). Moreover, the dietary addition of 200 mg/kg Bacillus coagulans also significantly enhanced the levels of interferon alpha (IFNα), toll-like receptor (TLR3), and melanoma differentiation-associated protein 5(MDA5) in the duodenum (p<0.05). In conclusion, the dietary addition of Bacillus coagulans significantly improved broiler performance, and enhanced the intestinal mucosal barrier and immune function. The optimal dosage of Bacillus coagulans for yellow broilers was determined as 2×108 cfu/kg.

ABSTRACT The objective of this work was to investigate the effect of feeding conditions on methylation status of FATP1 gene, which is an important candidate gene of Intramuscular fat and important indicator of chicken meat quality. We selected Daninghe (DNH) and Qingjiaoma (QJM) chickens under scatter-feeding and captivity-feeding conditions as experimental animals, and detected the methylation status of FATP1 genes in chicken breast muscle using Bisulfite Sequencing PCR method. The results showed that the methylation level of FATP1 in scatter-fed chicken was lower than in captivity-fed conditions in DNH and QIM chicken breast tissues; DNA methylation in the promoter and exon1 region was demonstrated to negatively regulate the expression of the FATP1 gene. These results suggested that feeding conditions affect the methylation status and expression level of FATP1, thereby affecting the Intramuscular fat content in DNH and QJM chicken breast muscle.

ABSTRACT Aim: To investigate the radiographic anatomical characters of the bones in the fractured calcaneus by using tangential X-rays. Methods: Fifty-seven patients (114 sides) with normal calcaneus and thirty patients (42 sides) with fractured calcaneus were radiographed along the line connecting the midpoint of the 4th and 5th metatarsal heads with the lateral margin of the corpus calcanei. The anatomical characters of the calcaneus on the tangential X-ray views were analysed, including radius and fractal geometry. Results: On a tangential X-ray view of the fractured calcaneus, the projection of sustentaculum tali resembled a duck beak, which was just under the middle facet of the talocalcaneal joint. The corpus calcanei appeared oval, which was just under the talocalcaneal joint articular surface. The lateral calcaneal wall did not exceed the vertical line extending from medial of the lateral malleolar fossa (the Z-W line). From the scan picture, the fractured calcaneus seemed to be widened as its lateral wall crossed the Z-W line. The longitudinal axis of this oval-shaped projection was 3.52 ± 0.41 cm (a), the horizontal axis was 2.56 ± 0.28 cm (b), and the a/b ratio was 1.40 ± 0.11. In the normal calcaneus, the a/b ratio is higher than 1.40 and the differences of the values in these two groups were significant. Therefore, the a/b ratio of the calcaneal oval-shaped projection can be used to evaluate the degree of its deformity after fracture. Conclusion: These results demonstrated that the tangential X-ray view of the calcaneus can be used to evaluate its radiographic anatomy and to measure the radii of the oval-shaped corpus calcanei.

RESUMEN Objetivo: Investigar las características anátomo-radiográficas de los huesos en el calcáneo fracturado, mediante el uso de rayos x tangenciales. Métodos: Cincuenta y siete pacientes (114 lados) con calcáneo normal y treinta pacientes (42 lados) con fractura del calcáneo fueron radiografiados a lo largo de la línea que une el punto medio de la 4ta y 5ta cabezas metatarsianas con el margen lateral del cuerpo calcáneo (corpus calcanei). Se analizaron los rasgos anatómicos del calcáneo observados en los rayos x tangenciales de rayos x, incluyendo el radio y la geometría fractal. Resultados: En la vista tangencial de la radiografía del calcáneo fracturado, la proyección del sustentaculum tali, se asemejó a un pico de pato, justamente debajo de la cara media de la articulación talocalcáne a. El cuerpo calcáneo de apariencia oval, se hallaba justo debajo de la superficie articular de la articulación talocalcánea. La pared lateral calcánea no excedía la línea vertical que se extiende desde la fosa maleolar medial a la lateral (línea Z-W). En la imagen escaneada, el calcáneo fracturado parecía haberse anchado cuando su pared lateral cruzó la línea Z-W. El eje longitudinal de esta proyección ovalada medía 3.52 ± 0.41 cm (a), el eje horizontal medía 2.56 ± 0.28 cm (b), y el cociente de a/b fue 1.40 ± 0.11. En el calcáneo normal, el cociente a/b es mayor de 1.40, y las diferencias de los valores en estos dos grupos fueron significativas. Por lo tanto, el cociente a/b de la proyección ovalada del calcáneo, puede usarse para evaluar su grado de deformidad después de la fractura. Conclusión: Estos resultados demostraron que la radiografía tangencial del calcáneo puede usarse para evaluar su anatomía radiográfica y medir los radios del cuerpo calcáneo ovalado.

The aim of this study was to investigate whether exogenous retinoic acid (RA) can upregulate the mRNA and protein expression of growth-associated protein 43 (GAP-43), thereby promoting brain functional recovery in a rat distal middle cerebral artery occlusion (MCAO) model of ischemia. A total of 216 male Sprague Dawley rats weighing 300–320 g were divided into 3 groups: sham-operated group, MCAO+vehicle group and MCAO+RA group. Focal cortical infarction was induced with a distal MCAO model. The expression of GAP-43 mRNA and protein in the ipsilateral perifocal region was assessed using qPCR and immunocytochemistry at 1, 3, 7, 14, 21, and 28 days after distal MCAO. In addition, an intraperitoneal injection of RA was given 12 h before MCAO and continued every day until the animal was sacrificed. Following ischemia, the expression of GAP-43 first increased considerably and then decreased. Administration of RA reduced infarction volume, promoted neurological functional recovery and upregulated expression of GAP-43. Administration of RA can ameliorate neuronal damage and promote nerve regeneration by upregulating the expression of GAP-43 in the perifocal region after distal MCAO.

Different pathways act synergistically to participate in many biological processes. Thus, the purpose of our study was to extract dysregulated pathways to investigate the pathogenesis of colorectal cancer (CRC) based on the functional dependency among pathways. Protein-protein interaction (PPI) information and pathway data were retrieved from STRING and Reactome databases, respectively. After genes were aligned to the pathways, each pathway activity was calculated using the principal component analysis (PCA) method, and the seed pathway was discovered. Subsequently, we constructed the pathway interaction network (PIN), where each node represented a biological pathway based on gene expression profile, PPI data, as well as pathways. Dysregulated pathways were then selected from the PIN according to classification performance and seed pathway. A PIN including 11,960 interactions was constructed to identify dysregulated pathways. Interestingly, the interaction of mRNA splicing and mRNA splicing-major pathway had the highest score of 719.8167. Maximum change of the activity score between CRC and normal samples appeared in the pathway of DNA replication, which was selected as the seed pathway. Starting with this seed pathway, a pathway set containing 30 dysregulated pathways was obtained with an area under the curve score of 0.8598. The pathway of mRNA splicing, mRNA splicing-major pathway, and RNA polymerase I had the maximum genes of 107. Moreover, we found that these 30 pathways had crosstalks with each other. The results suggest that these dysregulated pathways might be used as biomarkers to diagnose CRC.

15N tracing technique was applied to investigate the effects of various organic carbon (OC) sources on dissimilatory nitrate (NO3-)reduction to ammonium (NH4+) (DNRA) rates in the coastal wetland sediments. Soils collected from the Chongming Dongtan wetland were incubated at 25ºC in the dark for 24 h following the additions of OC sources (glucose, acetate, malate, citrate and oxalate (500 μg C g-1 dry soil)) and 15N-labeled NH4(15)NO3 (initial 15N atom% of NO3--N is 20%). The results showed that soil DNRA rates varied from 0.018-0.497 mg N kg-1 dry soil d-1during the whole incubation, and the rates differed significantly among treatments following the order: oxalate> citrate> glucose> acetate> malate> no exogenous C addition over the first 12-h incubation. This was possibly caused by the different decomposition rates of various OC sources, which further influenced the available energy provided for DNRA microorganisms. Soils with no addition of exogenous C showed lower soil DNRA rates, presumably because of the low C/NO3- ratio as well as energy availability. The relative lower soil DNRA rates over the 24-h incubation indicated that DNRA is a fast process. Our results suggest that DNRA could be controlled by OC sources, especially organic acids, demonstrating that the widespread use of glucose in soil laboratory studies might lead to misleading knowledge on understanding the effects of OC on soil DNRA process.

Pancreatic ductal adenocarcinoma (PDAC) has a poor 5-year survival rate of 5%. Biomarkers for the early detection of pancreatic cancer are urgently needed. Transforming growth factor-beta1 (TGF-β1) is elevated in the tissues and plasma of patients with PDAC. However, no studies systemically report prognostic significance of plasma TGF-β1 levels in PDAC. In the present study, we assessed the prognostic significance of serum TGF-β levels in patients with PDAC. TGF-β levels were determined in serum from 146 PDAC patients, and 58 patients with benign pancreatic conditions. Regression models were used to correlate TGF-β levels to gender, age, stage, class, and metastasis. Survival analyses were performed using multivariate Cox models. Serum levels of TGF-β1 distinguished PDAC from benign pancreatic conditions (P<0.001) and healthy control subjects (P<0.001). Serum levels of TGF-β also distinguished tumor stage (P=0.002) and lymph node metastasis (P=0.001). High serum levels of TGF-β1 were significantly correlated with reduced patient survival. Multivariate analysis revealed that TGF-β1, lymph node metastasis and tumor stage were independent factors for PDAC survival. Our results indicate that serum TGF-β1 may be used as a potential prognostic marker for PDAC.

We evaluated the effect of puerarin on spatial learning and memory ability of mice with chronic alcohol poisoning. A total of 30 male C57BL/6 mice were randomly divided into model, puerarin, and control groups (n=10 each). The model group received 60% (v/v) ethanol by intragastric administration followed by intraperitoneal injection of normal saline 30 min later. The puerarin group received intragastric 60% ethanol followed by intraperitoneal puerarin 30 min later, and the control group received intragastric saline followed by intraperitoneal saline. Six weeks after treatment, the Morris water maze and Tru Scan behavioral tests and immunofluorescence staining of cerebral cortex and hippocampal neurons (by Neu-N) and microglia (by Ib1) were conducted. Glutamic acid (Glu) and gamma amino butyric acid (GABA) in the cortex and hippocampus were assayed by high-performance liquid chromatography (HPLC), and tumor necrosis factor (TNF)-α and interleukin (IL)-1β were determined by ELISA. Compared with mice in the control group, escape latency and distance were prolonged, and spontaneous movement distance was shortened (P<0.05) by puerarin. The number of microglia was increased in both the cortex and hippocampal dentate gyrus (P<0.01), and neurons were reduced only in the hippocampal dentate gyrus (P<0.01) in puerarin-treated mice. In the model group, Glu and GABA levels decreased (P<0.05), and Glu/GABA, TNF-α, and IL-1β increased (P<0.01) with puerarin treatment, returning to near normal levels. In conclusion, puerarin protected against the effects of chronic alcohol poisoning on spatial learning and memory ability primarily because of anti-inflammatory activity and regulation of the balance of Glu and GABA.