Dietary supplementation with dihydroartemisinin improves intestinal barrier function in weaned piglets with intrauterine growth retardation by modulating the gut microbiota.

The aim of this study was to investigate whether dietary dihydroartemisinin (DHA) supplementation could improve intestinal barrier function and microbiota composition in intrauterine growth restriction (IUGR) weaned piglets. Twelve normal birth weight (NBW) piglets and 24 IUGR piglets at 21 d of age were divided into three groups, which were fed a basal diet (NBW-CON and IUCR-CON groups) and an 80 mg/kg DHA diet (IUGR-DHA group). At 49 d of age, eight piglets of each group with similar body weights within groups were slaughtered, and serum and small intestine samples were collected. The results showed that IUGR piglets reduced growth performance, impaired the markers of intestinal permeability, induced intestinal inflammation, decreased intestinal immunity, and disturbed the intestinal microflora. Dietary DHA supplementation increased average daily gain, average daily feed intake, and body weight at 49 d of age in IUGR-weaned piglets (P < 0.05). DHA treatment decreased serum diamine oxidase activity and increased the numbers of intestinal goblet cells and intraepithelial lymphocytes, concentrations of jejunal mucin-2 and ileal trefoil factor 3, and intestinal secretory immunoglobin A and immunoglobin G (IgG) concentrations of IUGR piglets (P < 0.05). Diet supplemented with DHA also upregulated mRNA abundances of jejunal IgG, the cluster of differentiation 8 (CD8), major histocompatibility complex-I (MHC-I), and interleukin 6 (IL-6) and ileal IgG, Fc receptor for IgG (FcRn), cluster of differentiation 8 (CD4), CD8, MHC-I, IL-6 and tumor necrosis factor α (TNF-α), and enhanced mRNA abundance and protein expression of intestinal occludin and ileal claudin-1 in IUGR piglets (P < 0.05). In addition, DHA supplementation in the diet improved the microbial diversity of the small intestine of IUGR piglets and significantly increased the relative abundance of Actinobacteriota, Streptococcus, Blautia and Streptococcus in the jejunum, and Clostridium sensu_ stricto_in the ileum (P < 0.05). The intestinal microbiota was correlated with the mRNA abundance of tight junction proteins and inflammatory response-related genes. These data suggested that DHA could improve the markers of intestinal barrier function in IUGR-weaned piglets by modulating gut microbiota. DHA may be a novel nutritional candidate for preventing intestinal dysfunction in IUGR pigs.

Intrauterine growth retardation (IUGR) is defined as the restricted development of the mammalian fetus or its organs during pregnancy, which has high morbidity and mortality during the perinatal period and improves the risk of metabolic diseases in the long term. Dihydroartemisinin (DHA) is a derivative of artemisinin that possesses anti-inflammatory and immunoregulatory effects. Therefore, this experiment was conducted to investigate whether dietary DHA supplementation could improve the intestinal barrier function and microbiota composition in IUGR-weaned piglets. The result showed that IUGR could lead to intestinal barrier dysfunction. Dietary supplementation with DHA improved growth performance and attenuated intestinal barrier dysfunction by decreasing the markers of intestinal permeability, increasing the mucus layer barrier, enhancing immunity, and reducing the inflammatory response in IUGR piglets, which may be attributed to the improvement of the intestinal microbiota. Moreover, the study indicated that the gut microflora was correlated with the gene expression of tight junction proteins and immune function. This study may provide a new nutritional strategy for the maintenance of intestinal health in IUGR pigs.

Effects of bamboo leaf flavonoids on growth performance, antioxidants, immune function, intestinal morphology, and cecal microbiota in broilers.

BACKGROUND: Bamboo leaf flavonoids (BLF) are the main bioactive ingredients in bamboo leaves. They have antioxidant, anti-inflammatory, antibacterial, and other effects. In this study, the effects of dietary BLF on growth performance, immune response, antioxidant capacity, and intestinal microbiota of broilers were investigated. A total of 288 broilers were divided into three groups with eight replicates and 12 birds in each replicate. Broilers were fed a basic diet or the basic diet supplemented with 1000 or 2000 mg kg-1 BLF for 56 days. RESULTS: The results showed that supplementation of BLF increased body weight (BW) and average daily weight gain (ADG), and reduced average daily feed intake (ADFI) (P < 0.05). The serum immunoglobulin A (IgA), immunoglobulin M (IgM), and interleukin 10 (IL-10) content of broilers in the BLF1000 group was increased and the interleukin 1ß (IL-1ß) and tumor necrosis factor-α (TNF-α) content was decreased (P < 0.05). The levels of IgM and IL-10 in jejunum mucosa were found to be enhanced by BLF (P < 0.05). The BLF1000 group exhibited a significant reduction in the concentration of TNF-α (P < 0.05). Serum and jejunum mucosa total antioxidant capacity (T-AOC) levels in the BLF1000 group were increased (P < 0.05). The serum catalase (CAT) and glutathione peroxidase (GSH-Px) effects of the BLF1000 group and serum CAT effects of BLF2000 group were increased (P < 0.05). The CON group demonstrated a lower relative abundance of Christensenellaceae_R-7_group and Oscillibacter than the BLF group (P < 0.05). CONCLUSION: Dietary BLF inclusion enhanced the growth performance, immune, and antioxidant functions, improved the intestinal morphology, and ameliorated the intestinal microflora structure in broiler. Adding 1000 mg kg-1 BLF to the broiler diet can be considered as an effective growth promoter. © 2024 Society of Chemical Industry.

Effects of three different plant-derived polysaccharides on growth performance, immunity, antioxidant function, and cecal microbiota of broilers.

BACKGROUND: This study investigated the effects of dietary plant polysaccharides on growth performance, immune status and intestinal health in broilers. We randomly divided 960 one-day-old Arbor Acres broiler chicks into four groups. The control (CON) group was fed a basal diet, and the remaining groups were fed a basal diet supplemented with 1000 mg kg-1 Ginseng polysaccharide (GPS), Astragalus polysaccharide (APS), or Salvia miltiorrhiza polysaccharide (SMP) for 42 days. RESULTS: Dietary supplementation with SMP significantly increased body weight (BW) at 21 and 42 days of age, average daily gain (ADG) and average daily feed intake (ADFI) during the starter and whole experimental period, decreased the concentrations of interleukin-1 beta (IL-1ß), tumor necrosis factor α (TNF-α) and malondialdehyde (MDA), increased the levels of interleukin-4 (IL-4) and interleukin-10 (IL-10) and catalase (CAT) activity in the serum (P < 0.05). GPS, APS, and SMP supplementation increased serum levels of immunoglobulins, activities of glutathione peroxidase (GSH-Px), total superoxide dismutase (T-SOD) and total antioxidant capacity (T-AOC), and cecal concentrations of acetic acid and propionic acid of broilers (P < 0.05). Furthermore, high-throughput sequencing results showed that the relative abundance of Firmicutes was decreased while the relative abundance of Bacteroidota, Alistipes, and Prevotellaceae_NK3B31_group were increased (P < 0.05) in the GPS, APS, and SMP groups compared with the CON group. CONCLUSION: Dietary GPS, APS, and SMP supplementation could improve growth performance, enhance immune function by increasing serum immunoglobulin and regulating cytokines, improve antioxidant function by increasing serum antioxidant enzyme activity, increase volatile fatty acid levels and improve the microbial composition in the cecum of broilers. Dietary SMP supplementation had the optimal effect in this study. © 2023 Society of Chemical Industry.

Plasma non-transferrin-bound iron uptake by the small intestine leads to intestinal injury and intestinal flora dysbiosis in an iron overload mouse model and Caco-2 cells.

Iron overload often occurs during blood transfusion and iron supplementation, resulting in the presence of non-transferrin-bound iron (NTBI) in host plasma and damage to multiple organs, but effects on the intestine have rarely been reported. In this study, an iron overload mouse model with plasma NTBI was established by intraperitoneal injection of iron dextran. We found that plasma NTBI damaged intestinal morphology, caused intestinal oxidative stress injury and reactive oxygen species (ROS) accumulation, and induced intestinal epithelial cell apoptosis. In addition, plasma NTBI increased the relative abundance of Ileibacterium and Desulfovibrio in the cecum, while the relative abundance of Faecalibaculum and Romboutsia was reduced. Ileibacterium may be a potential microbial biomarker of plasma NTBI. Based on the function prediction analysis, plasma NTBI led to the weakening of intestinal microbiota function, significantly reducing the function of the extracellular structure. Further investigation into the mechanism of injury showed that iron absorption in the small intestine significantly increased in the iron group. Caco-2 cell monolayers were used as a model of the intestinal epithelium to study the mechanism of iron transport. By adding ferric ammonium citrate (FAC, plasma NTBI in physiological form) to the basolateral side, the apparent permeability coefficient (Papp) values from the basolateral to the apical side were greater than 3×10-6 cm s-1. Intracellular ferritin level and apical iron concentration significantly increased, and SLC39A8 (ZIP8) and SLC39A14 (ZIP14) were highly expressed in the FAC group. Short hairpin RNA (shRNA) was used to knock down ZIP8 and ZIP14 in Caco-2 cells. Transfection with ZIP14-specific shRNA decreased intracellular ferritin level and inhibited iron uptake. These results revealed that plasma NTBI may cause intestinal injury and intestinal flora dysbiosis due to the uptake of plasma NTBI from the basolateral side into the small intestine, which is probably mediated by ZIP14.

Comparison of the Effects of Yucca saponin, Yucca schidigera, and Quillaja saponaria on Growth Performance, Immunity, Antioxidant Capability, and Intestinal Flora in Broilers.

The purpose of this study is to investigate the effects of Yucca saponin (YSa), Yucca schidigera (YS), and Quillaja Saponaria (QS) on growth performance, nitrogen metabolism, immune ability, antioxidant capability, and intestinal flora of yellow-feather broilers. This study randomly divided a total of 480 1-day yellow-feather broilers into 4 treatment groups. Factors in the 4 groups included CON group (basic diet), YSa group (basic diet mixed with 500 mg/kg YSa), YS group (basic diet mixed with 500 mg/kg YS), and QS group (basic diet mixed with 500 mg/kg QS). Throughout the 56-day study period, YSa, YS, and QS groups had higher average daily gain in broilers than the CON group (p < 0.01). The YS group had a lower feed gain ratio (F: G) in broilers than the CON group (p < 0.05). YSa, YS, and QS showed increased serum immunoglobin A (IgA), immunoglobin Y (IgY), immunoglobin M (IgM), and total antioxidant capacity (T-AOC) levels; enhanced acetic acid, butyric acid, and valeric acid levels of cecal content; and reduced contents of ammonia nitrogen, urea nitrogen, interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and malondialdehyde (MDA) in serum in broilers (p < 0.05). The relative abundance of Lachnoclostridium in the QS group was decreased compared with that in the CON group (p < 0.05). Higher IgA and IgY sera contents were observed in the YS group compared to the YSa and QS groups (p < 0.05). In contrast with the QS group, the serum IL-6 concentration of the YS group was reduced (p < 0.05). In conclusion, YSa, YS, and QS promoted growth performance, nitrogen metabolism, immunity, antioxidant capability, and intestinal flora in broilers. Through the comparison of YSa, YS, and QS, it was found that YS is more suitable as a feed additive to ameliorate the healthy growth of broilers.

Lactobacillus plantarum postbiotics trigger AMPK-dependent autophagy to suppress Salmonella intracellular infection and NLRP3 inflammasome activation.

We previously found that Lactobacillus plantarum (LP)-derived postbiotics protected animals against Salmonella infection, but the molecular mechanism remains obscure. This study clarified the mechanisms from the perspective of autophagy. Intestinal porcine epithelial cells (IPEC-J2) were pretreated with LP-derived postbiotics (the culture supernatant, LPC; or heat-killed bacteria, LPB), and then challenged with Salmonella enterica Typhimurium (ST). Results showed that LP postbiotics markedly triggered autophagy under ST infection, as indicated by the increased LC3 and Beclin1 and the decreased p62 levels. Meanwhile, LP postbiotics (particularly LPC) exhibited a strong capacity of inhibiting ST adhesion, invasion and replication. Pretreatment with the autophagy inhibitor 3-methyladenine (3-MA) led to a significant decrease of autophagy and the aggravated infection, indicating the importance of autophagy in LP postbiotics-mediated Salmonella elimination. LP postbiotics (especially LPB) significantly suppressed ST-induced inflammation by modulating inflammatory cytokines (the increased interleukin (IL)-4 and IL-10, and decreased tumor necrosis factor-α (TNF), IL-1ß, IL-6 and IL-18). Furthermore, LP postbiotics inhibited NOD-like receptor protein 3 (NLRP3) inflammasome activation, as evidenced by the decreased levels of NLRP3, Caspase-1 and apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC). Deficits in autophagy resulted in an increase of inflammatory response and inflammasome activation. Finally, we found that both LPC and LPB triggered AMP-activated protein kinase (AMPK) signaling pathway to induce autophagy, and this was further confirmed by AMPK RNA interference. The intracellular infection and NLRP3 inflammasome were aggravated after AMPK knockdown. In summary, LP postbiotics trigger AMPK-mediated autophagy to suppress Salmonella intracellular infection and NLRP3 inflammasome in IPEC-J2 cells. Our findings highlight the effectiveness of postbiotics, and provide a new strategy for preventing Salmonella infection.

Synthesis and anticancer activity of novel histone deacetylase inhibitors that inhibit autophagy and induce apoptosis.

The combination of histone deacetylase (HDAC) and autophagy inhibitor has been considered as a novel cancer therapeutic strategy. To find novel HDAC inhibitors that can inhibit autophagy, several new series of oxazole- and thiazole-based HDAC inhibitors were designed and synthesized by replacing the phenyl cap in SAHA with 5-phenyloxazoles and 5-phenylthiazoles. The representative oxazole derivative, compound 21, showed better enzymatic inhibitory activity than SAHA (vorinostat). Compound 21 induced G2/M cell cycle arrest and its antiproliferative activity is 10-fold better than SAHA in multiple tumor cell lines. Western blot analysis showed that compound 21 can markedly increase the acetylation levels of tubulin, histone H3, and histone H4. Contrary to SAHA, compound 21 was found to inhibit autophagy. Additionally, compound 21 induced cell apoptosis via the Bax/Bcl-2 and caspase-3 pathways. Ultimately, compound 21 exhibited higher oral antitumor potency than SAHA in a A549 xenograft model. Our results indicated that compound 21 may be further developed as a promising anticancer agent.

Rhamnolipids Regulate Lipid Metabolism, Immune Response, and Gut Microbiota in Rats.

Objectives: Gut microbes influence lipid metabolism and immune responses that are key features of metabolic disorders. This study examined effects of bacterial rhamnolipids (RLS) on lipid metabolism, immune response, and gut microbiota in rats. Methods: Twenty-four Sprague-Dawley rats were randomly divided into three groups and gavage-fed for seven weeks with normal saline (NCO group), 50 mg/kg bw RLS (RLS1 group), and 100 mg/kg bw RLS (RLS2 group). Results: Compared with those of the NCO group, the RLS1 and RLS2 groups showed significantly decreased fat weight, relative fat weight, and adipocyte size (P < 0.05). Furthermore, RLS1 and RLS2 significantly decreased concentrations of triglycerides, low-density lipoprotein-cholesterol, and non-esterified fatty acids and increased high-density lipoprotein-cholesterol levels (P < 0.05). However, the total cholesterol content among the three groups (P > 0.05) were not significantly different. Serum concentrations of interleukin-1ß, interleukin-6, and tumor necrosis factor-α were significantly lower in the RLS2 group than those in the NCO group (P < 0.05). The relative mRNA expression of fatty acid synthase was significantly decreased, while those of carnitine palmitoyltransferase-1, carnitine palmitoyltransferase-2, and peroxisome proliferator-activated receptor-gamma coactivator-1α were significantly increased in the RLS2 group compared with those in the NCO group (P < 0.05). Moreover, the relative abundances of Lactobacillus, Roseburia, Ruminococcus-1, and Parabacteroides were significantly higher in the RLS2 group than those in the NCO group (P < 0.05). Conclusion: Our findings suggest that RLS reduces fat deposition, inhibits inflammation, regulates intestinal flora, and promotes the proliferation of beneficial bacteria in rats.

Effects of dietary glyceryl monolaurate supplementation on growth performance, non-specific immunity, antioxidant status and intestinal microflora of Chinese mitten crabs.

This study aims to investigate the effects of glycerol monolaurate (GML) on growth performance, non-specific immunity, antioxidant capacity and intestinal microflora in Chinese mitten crabs. The crabs were randomly arranged to three experimental diets groups containing 0 (control group), 1000 mg/kg GML (GML1000 group), and 2000 mg/kg GML (GML2000 group), respectively. After 8 weeks of breeding, results showed a better growth performance in GML2000 group, with a higher PWG, SGR and lower FCR (P < 0.05). Meanwhile, in GML2000 group the activities of phenoloxidase, alkaline phosphatase, acid phosphatase and lysozyme in hemolymph were increased (P < 0.05), also the activities of hemolymph and hepatopancreas superoxide dismutase (SOD) and glutathione peroxidase (GPx) were increased in hepatopancreas (P < 0.05). While malondialdehyde (MDA) concentrations were lower significantly (P < 0.05) both in GML1000 and GML2000 groups. Furthermore, the mRNA expression of TLR1, TLR2, which related to the Toll pathway were increased (P < 0.05). Supplementation of 2000 mg/kg GML up-regulated the expression of ALF and LZM (P < 0.05), and down-regulated the expression of caspase-3 (P < 0.05). The abundance of Firmicutes increased in GML2000 group (P < 0.05), and Shewanella was significantly increased (P < 0.05) in both GML1000 and GML2000 groups. In conclusion, dietary supplemented with GML enhanced the growth performance and antioxidant capacity, enhanced hemolymph immune enzymes activities and antimicrobial peptides expression through regulating the proPO system and Toll pathway, and improved gut microflora in Chinese mitten crabs.

Multi-Omics Revealed the Protective Effects of Rhamnolipids in Lipopolysaccharide Challenged Broilers.

Rhamnolipid (RL) is a glycolipid biosurfactant and exhibits the following outstanding characteristics: strong antibacterial properties, low toxicity, and high biodegradability. The present research was conducted to explore the protective effects and mechanisms of rhamnolipids as an alternative to antibiotics in LPS (lipopolysaccharide)-challenged broilers. 16S rRNA gene sequencing and metabolomics were used for analyzing the cecal microbial composition and serum metabolites. Dietary antibiotics and RLS supplementation decreased the weight loss rate, enhanced serum immunoglobulin levels, reduced serum diamine oxidase and D-lactate acid concentration, and improved the symptoms of intestinal bleeding and villus height, when broilers were challenged with LPS. The addition of RLS in the diet enhanced serum interleukin-4 and interleukin-10 contents and reduced serum interleukin-6 and tumor necrosis factor-α levels in LPS-challenged broilers compared with the antibiotics group. Spearman's correlation analysis revealed that RLS may alleviate LPS-induced inflammatory responses through altering the 6-methoxymellein level in broilers. The genus Bacteroides may contribute to the decreased weight loss rate via regulating the serum lysoPC [20:5(5Z,8Z,11Z,14Z,17Z)] secretion. RLS alleviates LPS-induced intestinal injury, enhances the growth and immunity, ameliorates intestinal microflora, and improves serum metabolites in LPS-challenged broilers. RLS exhibited better protective effect than antibiotic supplementation in the diet of LPS-challenged broilers. These findings provide potential regulation strategies and novel insights for RLS enhancing its protective effect in LPS-challenged broilers.

Combined screening analysis of aberrantly methylated-differentially expressed genes and pathways in hepatocellular carcinoma.

Background: Methylation plays an important role in hepatocellular carcinoma (HCC) by altering the expression of key genes. The aim of this study was to screen the aberrantly methylated-differentially expressed genes (DEGs) in HCC and elucidate their underlying molecular mechanism. Methods: Gene expression microarrays (GSE101685) and gene methylation microarrays (GSE44909) were selected. DEGs and differentially methylated genes (DMGs) were screened. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were performed using the Database for Annotation, Visualization, and Integrated discovery (DAVID). The Search Tool for the Retrieval of Interacting Genes (STRING) database was used to analyze the functional protein-protein interaction (PPI) network. Molecular Complex Detection (MCODE) analysis was performed using the Cytoscape software. Hub genes were verified in The Cancer Genome Atlas (TCGA) database. Results: A total of 80 hypomethylation-high expression genes (Hypo-HGs) were identified. Pathway enrichment analysis showed DNA replication, cell cycle, viral carcinogenesis, and the spliceosome. The top 5 hub genes were minichromosome maintenance complex component 3 (MCM3), checkpoint kinase 1 (CHEK1), kinesin family member 11 (KIF11), PDZ binding kinase (PBK), and Rac GTPase activating protein 1 (RACGAP1). In addition, 189 hypermethylation-low expression genes (Hyper-LGs) were identified. Pathway enrichment analysis indicated enrichment in metabolic pathways, drug metabolism-other enzymes, and chemical carcinogenesis. The top 5 hub genes were leukocyte immunoglobulin like receptor B2 (LILRB2), formyl peptide receptor 1 (FPR1), S100 calcium binding protein A9 (S100A9), S100 calcium binding protein A8 (S100A8), and myeloid cell nuclear differentiation antigen (MNDA). The methylation status and mRNA expression of MCM3, CHEK1, KIF11, PBK, and S100A9 were consistent in the TCGA database and significantly correlated with the prognosis of patients. Conclusions: Combined screening of aberrantly methylated-DEGs based on bioinformatic analysis may provide new clues for elucidating the epigenetic mechanism in HCC. Hub genes, including MCM3, CHEK1, KIF11, PBK, and S100A9, may serve as biomarkers for the precise diagnosis of HCC.

Garlic Powder Supplementation Improves Growth, Nonspecific Immunity, Antioxidant Capacity, and Intestinal Flora of Chinese Mitten Crabs (Eriocheir sinensis).

This study was conducted to survey the effects of garlic powder on growth performance, nonspecific immunity, antioxidant capacity, and intestinal flora structure of Chinese mitten crabs. Altogether, 216 crabs which originally weigh 20.71 ± 0.13 g were randomly allocated into three treatment groups with 6 replicates of 12 crabs per replicate. The control group (CN) was fed a basal diet, while the other two groups were fed the basal diet supplemented with 1000 mg/kg (GP1000) and 2000 mg/kg (GP2000) garlic powder, respectively. This trial lasted 8 weeks. The results showed that the supplementation of garlic powder improved the final body weight, weight gain rate, and specific growth rate of the crabs (P < 0.05). Meanwhile, in serum, better nonspecific immune was confirmed by the enhancement of phenoloxidase and lysozyme levels, with the improvement of phosphatase activities in GP1000 and GP2000 (P < 0.05). On the other hand, the levels of total antioxidant capacity, glutathione peroxidases, and total superoxide dismutase in serum and hepatopancreas were increased (P < 0.05) while malondialdehyde content declined (P < 0.05) as the garlic powder was added to the basal diet. And, catalase in serum also shows an increase (P < 0.05). In both GP1000 and GP2000, genes related to antioxidant and immunity, for instance, Toll-like receptor 1, glutathione peroxidase, catalase, myeloid differentiation factor 88, TuBe, Dif, relish, crustins, antilipopolysaccharide factor, lysozyme, and prophenoloxidase mRNA expression levels, were increased (P < 0.05). The abundance of Rhizobium and Rhodobacter was reduced by adding garlic powder (P < 0.05). This study indicated that dietary addition of garlic powder promoted growth, enhanced nonspecific immunity and antioxidant capacity, activated Toll pathway, IMD pathway, and proPO system, increased antimicrobial peptide expression, while simultaneously improving the intestinal flora of Chinese mitten crabs.

Effects of Bacillus subtilis and Bacillus licheniformis on growth performance, immunity, short chain fatty acid production, antioxidant capacity, and cecal microflora in broilers.

This study investigated the effects of dietary supplementation with Bacillus subtilis (B. subtilis) or Bacillus licheniformis (B. licheniformis) on growth performance, immunity, antioxidant capacity, short chain fatty acid (SCFA) production, and the cecal microflora in broiler chickens. In total, 360 male, 1-day-old Cobb 500 birds were randomly divided into 3 groups: the control group was fed a basal diet; the B. subtilis group was fed a basal diet supplemented with 1.5 × 109 CFU/kg B. subtilis; the B. licheniformis group was fed a basal diet supplemented with 1.5 × 109 CFU/kg B. licheniformis. Results showed that chickens supplemented with either B. subtilis or B. licheniformis had comparatively higher (P < 0.05) body weight and average daily gain, whereas no difference (P > 0.05) was observed in feed efficiency. Concentrations of serum IgA, IgY, and IgM, as well as anti-inflammatory IL-10 were significantly increased (P < 0.05), and proinflammatory IL-1ß and IL-6 were significantly decreased (P < 0.05) by B. subtilis or B. licheniformis supplementation. Moreover, chickens fed with diets supplemented by either B. subtilis or B. licheniformis had greater antioxidant capacity, indicated by the notable increases (P < 0.05) in glutathione peroxidase, superoxide dismutase, and catalase, along with decrease (P < 0.05) in malondialdehyde. Compared to the control group, levels of SCFA, excluding acetic and propionic acid, in cecal content had improved (P < 0.05) by adding B. licheniformis, and significant increase (P < 0.05) in acetic and butyric acid was observed with B. subtilis supplementation. Microbial analysis showed that both B. subtilis or B. licheniformis supplementation could increase butyrate-producing bacteria such as Alistipes and Butyricicoccus, and decrease pathogenic bacteria such as the Synergistetes and Gammaproteobacteria. In summary, dietary supplemented with B. subtilis or B. licheniformis improved growth performance, immune status, and antioxidant capacity, increased SCFA production, and modulated cecal microbiota in chickens. Moreover, B. licheniformis was more effective than B. subtilis with the same supplemental amount.

Effects of Bacillus Coagulans on growth performance, antioxidant capacity, immunity function, and gut health in broilers.

This study evaluated the effects of Bacillus coagulans (B. coagulans) as an alternative to antibiotics on growth performance, antioxidant capacity, immunity function and gut health in broilers. A total of 480 one-day-old broilers were randomly divided into 3 treatments with 8 replicates comprising 20 broilers each. The experiment lasted 42 d. Treatments included: basal diet without antibiotics (NCO); basal diet supplemented with 75 mg/kg chlortetracycline (ANT); basal diet supplemented with 5 × 109 CFU/kg B. coagulans(BC). The B. coagulans enhanced body weight (BW) and average daily gain compared with the NCO group (P < 0.05). However, there were no significant differences in average daily feed intake and feed: gain ratio (F: G) among three groups (P > 0.05). The B. coagulans significantly increased catalase, superoxide dismutase, and glutathione peroxidase levels and reduced malondialdehyde levels (P < 0.05). The serum immunoglobulins (IgA, IgM, and IgY) were significantly higher in the BC group when compared to the NCO and ANT groups (P < 0.05). The B. coagulans also markedly reduced serum levels of proinflammatory factors (IL-1ß, IL-6, and TNF-α) and enhanced anti-inflammatory factor (IL-10) concentrations compared with control group (P < 0.05). Moreover, compared with the control group, BC significantly inhibited serum xanthine oxidase activity (P < 0.05). The levels of acetic acid, propionic acid, butyrate, isobutyric acid and valerate in BC group were significantly increased on d 42 compared with the NCO and ANT groups (P < 0.05). Furthermore, BC significantly altered cecal microbiota by reducing Desulfovibrio and Parasutterella, and by increasing Alistipes and Odoribacter (P < 0.05, P < 0.05, P < 0.001, P < 0.01, respectively). In conclusion, dietary B. coagulans, when used as an alternative to antibiotics, improved body weight, average daily gain, antioxidant capacity, immunity function and gut health in broilers.

Synthesis and structure-activity relationships of 5-phenyloxazole-2-carboxylic acid derivatives as novel inhibitors of tubulin polymerization.

A series of 5-phenyloxazole-2-carboxylic acid derivatives were synthesized, and their structure-activity relationships (SARs) were studied. N,5-diphenyloxazole-2-carboxamides 6, 7, and 9, which mimicked ABT751, showed improved cytotoxicity compared with ABT751. Compound 9 exhibited the highest antiproliferative activities against Hela A549, and HepG2 cancer cell lines, with IC50 values of 0.78, 1.08, and 1.27 µM, respectively. Furthermore, compound 9 showed selectivity for human cancer cells over normal cells, and this selectivity was greater than those of ABT751 and colchicine. Preliminary mechanism studies suggested that compound 9 inhibited tubulin polymerization and led to cell cycle arrest at G2/M phase. Molecular docking studies indicated that compound 9 bound to the colchicine binding site of tubulin. Our findings provided insights into useful SARs for further structural modification of inhibitors of tubulin polymerization.

Effects of dietary yeast extract supplementation on growth, body composition, non-specific immunity, and antioxidant status of Chinese mitten crab (Eriocheir sinensis).

This study was conducted to evaluate the effects of dietary supplementation with yeast extract on growth, body composition, non-specific immunity, and antioxidant status of Chinese mitten crab (Eriocheir sinensis). A total of 432 crabs (initial average weight, 4.62 ±â€¯0.11 g) were randomly distributed into four treatment groups with six replicates of 18 crabs. The crabs were fed a basal diet or the same diet supplemented with 2.5, 5, and 10 g/kg yeast extract for 8 weeks. The results showed that dietary yeast extract inclusion enhanced the edible viscera index (linear, P < 0.001), edible viscera crude protein (CP) content (linear, P = 0.025) and serum phenoloxidase (ProPO) activity (quadratic, P = 0.023) at 56 day, increased the total superoxide dismutase (T-SOD) activity at 28 day (quadratic, P = 0.037) and catelase (CAT) activity at 56 day (quadratic, P = 0.034) of edible viscera, and muscular T-SOD activity (quadratic, P = 0.020) at 56 day in Chinese mitten crab. Compared with the control group, the inclusion of 5 g/kg yeast extract in the diet increased the edible viscera index, enhanced the CAT activity of edible viscera at 56 day in Chinese mitten crab (P < 0.05). Dietary 10 g/kg yeast extract inclusion enhanced the edible viscera index at 56 day in Chinese mitten crab than that of the control group (P < 0.05). These results implied that dietary yeast extract inclusion improved the edible viscera index and crude protein content of edible viscera, enhanced serum immunity, and increased the antioxidant status of edible viscera and muscle in Chinese mitten crab, especially when it is supplemented at 5 g/kg yeast extract in the diet.

[Characteristics of recurrence of T1G3 transitional cell carcinoma of bladder].

OBJECTIVE: To investigate the characteristics of recurrence of T1G3 transitional cell carcinoma (TCC) of bladder. METHODS: The clinical data of 72 patients with TCC of bladder of the stage T1, 37 being of the stage T1G3 and 35 being of the stage T1G1-2, who underwent the initial treatment by transurethral resection of bladder tumor plus post-operative bladder infusion were analyzed. RESULTS: The first, second, third, and fourth year recurrent rates of the T1G3 group were 29.7%, 40.5%, 18.9%, and 10.8% respectively, all significantly higher than those of the T1G2 group (14.3%, 34.3%, 31.4%, and 20.0% respectively, t = 1.994, P = 0.025). The rate of recurrence for only time was 10.8% in the T1G3 group (4/37), significantly lower than that of the T1G1-2 group (17/35, 48.6%, P < 0.01). The rate of recurrence for more than 3 times was 54.0% in the T1G3 group (20/37), significantly higher than that of the T1G1-2 group (6/35, 17.2%, P < 0.01). The rate of recurrent tumor at the pathological stage over T2 was 86.5% (32/37) in the T1G3 group, higher than that of the T1G1-2 group (42.8%, 15/35). The rate of partial or total cystectomy at the last recurrence in the T1G3 group was 94.6% (35/37), significantly higher than that of the T1G1-2 group (57.1%, 20/35, chi(2) = 14.00, P < 0.05). CONCLUSION: A kind of dangerous bladder cancer, TCC at the stage T1G3 is apt recur soon after the treatment with a high recurrent frequency. Most recurrent tumors evolve into invasive cancer.

[Separation and purification of skeletal muscle satellite cells for tissue engineering applications by Percoll].

OBJECTIVE: To investigate separation and purification of skeletal muscle satellite cells with improved incontinuous density Percoll gradient centrifugation technique. METHODS: The primary skeletal muscle satellite cells of New Zealand white rabbits were cultured with different adhesion time method and incontinuous density Percoll gradient centrifugation technique. The cells were observed under invert microscope and scanning electron microscope. The degree of purification was examined by celluar immunochemical stain. The growth curve was tested by thiazolyl blue assay. RESULTS: Over 90% satellite cells were harvested by incontinuous density Percoll gradient centrifugation technique, in contrast to which, only 30%-40% cells were harvested by different adhesion time. Morphological observation accorded with satellite cells. The growth curve indicated that the cells grew in a good status. CONCLUSION: The high purification satellite cells can be obtained by incontinuous density Percoll gradient centrifugation technique. It is a good method to culture seed cells for tissue engineering applications.

[Preliminary research on preparation of porcine bladder acellular matrix graft for tissue engineering applications].

OBJECTIVE: To investigate the preparation method of porcine bladder acellular matrix graft (BAMG) and evaluate the feasibility of using BAMG as biomaterial scaffold to construct tissue engineering bladder. METHODS: Bladders were harvested from adult pigs at a slaughterhouse and were decellularized by the method of enzyme digestion or eradicator washing. The BAMG was then examined by optic microscope and electron microscope to confirm no cell elements remained. The structure of the BAMG was always observed through the Scanning Microscope. The skeletal muscle stem cells of New Zealand white rabbits were implanted in 96-hole-plank and cultured by BAMG extract liquid or normal culture medium. The cytotoxicity of BAMG was tested through MTT assay. New Zealand white rabbits were underwent bladder triangle area spared cystectomy and reconstructed by the heterogeneity BAMG. The animal bladders were obtained after six weeks and the tissue regeneration was examined. RESULTS: Good outcome were obtained with either of the two methods. There were no cell elements remained under the examination of optical microscope and electron microscope. The interstice could observed on BAMG through the Scanning Microscope. The relative growth rate (RGR) was 81%, 88%, 93% and 96% separately on the 2nd, the 4th, the 6th and the 8th day (t = 3.7419, P < 0.025). The cytotoxicity score was I, which indicated that BAMG had a good cytocompatibility. All rabbits were alive except one which died from diarrhea. The reconstructed bladder resumed the construction and function of the bladder. The transitional cells and muscle cells growth could be seen in the scaffold of the repaired bladder after six weeks. CONCLUSION: The porcine BAMG has good biocompatibility and can be used as ideal biomaterial scaffold of bladder tissue engineering.