Effect of Echinacea purpurea (L.) Moench and its extracts on the immunization outcome of avian influenza vaccine in broilers.

ETHNOPHARMACOLOGICAL RELEVANCE: Echinacea purpurea (L.) Moench (EP) is a perennial herbaceous flowering plant with immunomodulatory effects. However, the immunomodulatory effects of EP on broilers after vaccination are still unclear. AIM OF THE STUDY: The aim is to study the effect of EP and Echinacea purpurea (L.) Moench extracts(EE) on avian influenza virus (AIV) immunity, and further explore the potential mechanism of immune regulation. MATERIALS AND METHODS: Broilers were fed with feed additives containing 2% EP or 0.5% EE, and vaccinated against avian influenza. The samples were collected on the 7th, 21st, and 35th day after vaccination, and the feed conversion ratio (FCR) was calculated. Blood antibody titer, jejunal sIgA content, tight junction protein, gene and protein expression of TLR4-MAPK signaling pathway were also detected. RESULTS: The results showed that vaccination could cause immune stress, weight loss, increase sIgA content, and up-regulate the expression of tight junction proteins, including zonula occludens-1 (ZO-1), Occludin, and Claudin-1, as well as the genes of Toll-like receptor 4 (TLR4), myeloid differentiation primary response 88 (MyD88), receptor-associated factor 6 (TRAF6), activator protein 1 (AP-1) protein gene expression on TLR4-mitogen-activated protein kinase (MAPK) signaling pathway, and the protein expression of MyD88, extracellular regulated protein kinases (ERK), and c-Jun N-terminal kinase (JNK). EP and EE could increase the body weight of broilers, further improve antibody titers, decrease FCR, increase sIgA levels, up-regulate the expression of tight junction proteins, including ZO-1, Occludin, and Claudin-1, as well as the genes of TLR4, MyD88, TRAF6, and AP-1 and the protein expression of MyD88, ERK, and JNK in the TLR4-MAPK signaling pathway. CONCLUSION: In conclusion, EP and EE can increase the broiler's production performance and improve vaccine immune effect through the TLR4-MAPK signaling pathway.

Effect of Wu Zhi San supplementation in LPS-induced intestinal inflammation and barrier damage in broilers.

Intestinal inflammation and barrier damage can inhibit the absorption and transportation of nutrients in the small intestine, and lead to various chronic diseases. Wu Zhi San (WZS) is a traditional Chinese formula composed of Schisandrae, Anemarrhenae, Lonicerae, and Glycyrrhizae that was made to cure intestinal inflammation and barrier damage in broilers. To evaluate the protective effect of WZS on intestinal inflammation and barrier damage of broilers under lipopolysaccharide (LPS) stress, a total of 200 one-day-old broilers were randomly divided into five groups, namely, the CON group, LPS group, and three WZS groups (WZS-H, WZS-M, and WZS-L). The groups were designed for stress phase I (days 15, 17, 19, and 21) and stress phase II (days 29, 31, 33, and 35). The protective effect of WZS on the intestinal tract was evaluated by measuring the levels of serum myeloperoxidase (MPO), diamine oxidase (DAO), super oxide dismutase (SOD), and serum D-lactate (D-LA) and the expression of inflammatory factors in jejunum. The results showed that the diet supplemented with WZS could significantly reduce serum MPO, DAO, and D-LA levels and jejunal CD in broilers (p < 0.05), increase serum SOD levels and jejunal VH (p < 0.05), significantly downregulate the expression of NF-κB, TLR4, MyD88, and inflammatory cytokines (TNF-α, IL-1ß, IL-6, and IL-10), and upregulate Claudin-1, Occludin-1, and ZO-1 in broiler jejunum mucosa (p < 0.05). On the other hand, WZS could significantly reduce the protein expression of NF-κB (p65) in broiler jejunum (p < 0.05). These results indicate that supplementing WZS in the diet can reduce intestinal inflammation and alleviate intestinal barrier damage, and by inhibiting the NF-κB/TLR4/MyD88 signaling pathway, supplementation with WZS intervenes in LPS-induced stress injury in broilers.

The mixture of Radix isatidis, Forsythiae, and Gypsum alleviates lipopolysaccharide-induced fever in broilers by inhibition of TLR4/NF-κB signaling pathway.

To determine whether the antipyretic effect of the mixture of Radix isatidis, Forsythiae, and Gypsum (RIFG) on lipopolysaccharide (LPS) induced fever broilers and its related mechanisms. A total of 315 24-day-old yellow-plumed broilers were randomly divided into 7 groups, except for the control group, other groups were injected with LPS. Two hours later, RIFG were given drinking water to relieve fever, and it was evaluated by the expression of genes and proteins of the maximum body temperature rise (∆T), body temperature response index (TRI), serum and hypothalamic pyrogenic heat factor. RIFG could reduce the body temperature of broilers with fever (P < 0.01). It inhibited the expressions of IL-6 and PGE2 (P < 0.01), down-regulated mRNA expression levels of TNF-ɑ and COX-2 (P < 0.01), and promoted the generation of antipyretic factor AVP mRNA (P < 0.01). In addition, the expression level of TLR4 and NF-κB p65 protein can be down-regulated, and LPS + RM group has the best down-regulated effect. RIFG had a good antipyretic effect on reducing LPS-induced fever of broilers by inhibiting the activation of TLR4/NF-κB signaling pathway and thermogenic factors.

Echinacea purpurea (L.) Moench extract suppresses inflammation by inhibition of C3a/C3aR signaling pathway in TNBS-induced ulcerative colitis rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Echinacea purpurea (L.) Moench (EP) is a perennial herbaceous flowering plant, commonly known as purple conical flower. It was widely used to treat skin inflammation and gastrointestinal diseases. AIM OF STUDY: Ulcerative colitis (UC) is a chronic and nonspecific inflammatory disease. Recent evidence shows that immune disorders are involved in the pathogenesis of UC. To evaluate the protective effect of Echinacea purpurea (L.) Moench exact (EE) on UC and explore the role of complement system in the treatment of UC. MATERIALS AND METHODS: UC model was induced in rats by 2,4,6-trinitrobenzene sulfonic acid (TNBS), and then rats were administered with EE for 10 days. Collect colon tissues for analysis of relevant mechanisms. RESULTS: EE could reduce the weight loss and diarrhea of UC rats. In addition, EE could improve the integrity of intestinal epithelial barrier in UC rats. EE inhibited the level of proinflammatory cytokines and promoted the antioxidation. Furthermore, EE suppressed the expression of C3aR, CFB, CD55, TLR4 and NLRP3. CONCLUSION: These results indicate that EE may achieve therapeutic effect by inhibiting C3a/C3aR signal pathway, suggesting that EE may be used as a medicinal plant to alleviate UC.

Integrated gut microbiota and metabolomic analysis reveals immunomodulatory effects of Echinacea extract and Astragalus polysaccharides.

Immunosuppression in different animals increases the susceptibility of various infections caused by pathogenic microorganisms leading to increase risks posed by antibiotics in different animal farming sectors. Therefore, investigation of the interactions between natural medicines and the intestinal environmental ecosystem is of vital importance and crucial. This study for the first time investigated the effects of Echinacea Extract (EE) and Astragalus polysaccharide (APS) on the gut using 16S rRNA and metabolomic analysis approaches in immunosuppressed broiler chickens. There were four groups divided into control (C), immunosuppression (IS), EE, and APS groups. Sequencing of gut microbes showed that immunosuppression decreased the relative abundance of Anaerofustis, Anaeroplasma, Anaerotroncus, and Lachnospira in the gut while increasing that of c_115 and Holdemania. However, EE and APS diminished the effects on the immunosuppression on the microbiota. The results revealed up-regulation of the relative abundance of Enterococcus in broiler chickens. In addition, EE reduced the relative abundance of Ruminococcus and Blautia. The results on metabolomic analysis revealed that immunosuppression mainly affects cyanuric acid metabolism, starch and sucrose metabolism while interconversion of pentose and glucuronide. EE and APS, on the other hand mainly impact butyrate metabolism, alanine, aspartate and glutamate metabolism while the interconversion of pentose and glucuronide, and D-glutamine and D-glutamate metabolism. Results regarding correlation analysis revealed significantly metabolic pathways including TCA cycle, butyrate metabolism, glyoxylate and dicarboxylate metabolism, propionate metabolism, alanine, aspartate and glutamate metabolism associated with Ruminococcus and Blautia. Both EE and APS can antagonize the effects of immunosuppression by modulating the disrupted gut microbiota. Nevertheless, EE might have a bidirectional regulatory functions on the intestinal health and further studies are needed to know the exact and relevant mechanisms of action regarding the effects of EE and APS.

Effect of Traditional Chinese Medicine on the Gut Microbiota in Heat-Stressed Laying Hens.

Gut microbiota plays an important role in health and disease. To determine whether the traditional Chinese formula Zi Huang Huo Xiang San (ZHHXS) modulates gut microbiota under heat stress, a heat stress model was prepared in Roman layer hens by housing them at temperatures of 32-36°C and administering ZHHXS for 4 weeks. The Roman egg layers were randomly divided into three groups with 10 hens in each: a ZHHXS treatment group (ZHHXS-HS), a heat-stressed group (HS), and a blank control group (BC). The ZHHXS-HS and HS groups were housed in a 34 ± 2°C environment, while the BC group was housed at 25 ± 1°C. The ZHHXS-HS hens were fed a diet supplemented with 1% ZHHXS from 1 to 28 days, while the other groups were not. Gut microbiota in the hens' feces was assessed through 16S rRNA high-throughput sequencing on days 1, 3, 7, 14, and 28. A plot of the PCA scores showed that the gut microbiota composition in the BC group was a similar trend in the ZHHXS-HS group on days 1 and 3. The principal coordinate analysis (PCoA) unweighted distribution showed that the gut microbiota composition had no significant differences between the BC and ZHHXS-HS groups on days 1 and 7. The PCoA weighted distribution showed that the gut microbiota composition had no significant differences between the BC and ZHHXS-HS groups on days 1 and 3. This study showed that the composition of gut microbiota in layer hens with heat stress was modulated by ZHHXS treatment. ZHHXS treatment caused key phylotypes of gut microbiota to match the BC group, particularly Actinobacteria, Bacteroidetes, Bacteroides, and Enterococcus. The effect of ZHHXS in alleviating heat stress could be achieved by altering the composition of gut microbiota and regulating some key phylotypes.

Effect of Echinacea on gut microbiota of immunosuppressed ducks.

Introduction: Immunosuppression puts animals in a susceptible state and disrupts the balance of intestinal flora, which can increase the risk of disease and cause serious harm to the farm. Echinacea can exert its immunomodulatory effect in various ways, but its influence on intestinal flora is unclear. Methods: Therefore, we investigated the effect of Echinacea extract (EE) on gut microbiota in immunosuppressed ducks by 16s-RNA sequencing in this experiment. Results: The results showed that EE significantly improved the weight gain of immunosuppressed ducks (p<0.001). It also increased the immune organ index (p<0.01) and upregulated the levels of TNF-α and IFN-γ (p<0.05) as well as IL-2 in the serum. The lesions of the bursa were evident compared to the spleen and thymus. After treatment in the EE group, the lymphocyte count of the bursa returned to healthy levels and the lesions were significantly improved. The diversity analysis showed that neither of the alpha-diversity indices showed a significant difference (p>0.05). However, the EE group had a trend closer to the healthy group compared to the M group. ß-diversity analysis revealed a high degree of sample separation between the healthy and immunosuppressed groups. The sequencing result showed a significantly higher relative abundance of Prevotella and Prevotella_UCG_001 in the dexamethasone-treated group, which could be potential biomarkers of dexamethasone-induced immunosuppression. EE increased the relative abundance of Akkermansia, Bacteroides, and Alistipes and significantly decreased the relative abundance of Megamonas, Streptococcus, and Enterococcus (p<0.05). Conclusion: The results showed that Echinacea extract improves the development of immunosuppressed ducks and modulates intestinal immune function by increasing the abundance of beneficial bacterial genera in the intestine.

Antioxidant activities of sulfated Codonopsis polysaccharides in acute oxidative stress.

This study aimed to explore the protective effect of sulfated Codonopsis polysaccharides (SCP) on acute oxidative stress. SCP was modified by chlorosulfonic acid-pyridine method from Codonopsis polysaccharides (CP), which had 34.48% of sulfate content determined by ultrasonic-acidic barium chromate spectrophotometry. The analysis of Fourier transform-infrared spectroscopy (FT-IR) appeared an absorption peak of SCP at 811.91 cm-1 , which related to C-O-SO3 . In vitro test, the antioxidant activities of CP and SCP was induced by H2 O2 in RAW264.7 cells, results indicated that SCP and CP could significantly enhance the activity of superoxide dismutase (SOD), glutathione peroxidase (GDH-Px) and catalase (CAT), and nitric oxide (NO) and decrease the level of malondialdehyde (MDA), reactive oxygen species (ROS), and inducible nitric oxide synthase (iNOS) secreted by RAW264.7 cells compared with modeling group (p < .05). The flow cytometry results also revealed that SCP and CP could markedly inhibit the apoptosis of macrophage induced by acute oxidative stress. In vivo test, 50% ethanol was used to induce mice acute oxidative stress, results indicated that the blood biochemical parameters in mice were restored to normal levels following administration of SCP and CP, and alanine aminotransferase (ALT), aspartate transaminase (AST), total protein (TP), albumin (ALB), glucose (GLU), and creatinine (UREA) had significant differences compared with modeling group (p < .05). Quantitative real-time PCR analysis revealed that SCP and CP could promote the expression of Keap1 and Nrf2. In summary, both SCP and CP had protective effects against acute oxidative stress. PRACTICAL APPLICATIONS: Oxidative stress is a kind of stress injury, which can cause a variety of diseases and accelerate physical aging. Codonopsis has many active components, among which Codonopsis polysaccharide has antioxidant effect. Recent studies have found that Codonopsis polysaccharides could be modified by sulfate molecules to obtain higher antioxidant activity. The modified Codonopsis polysaccharides could significantly promote the production of antioxidant enzymes (SOD, CAT, GDH-Px) and reduce the content of oxidative stress marks (ROS, MDA). Moreover, its antioxidant mechanism may be related to the Keap1 /Nrf2 signaling pathway. Therefore, SCP was an effective antioxidant, and could be used as a potential health food with antioxidant and anti-aging effects.

Effects of ShenLing BaiZhu San Supplementation on Gut Microbiota and Oxidative Stress in Rats with Ulcerative Colitis.

The aim of this study was to evaluate the effect of gut microbiota and antioxidation of Shenling Baizhu San (SLBZS) as a supplement in a rat model of ulcerative colitis (UC) induced by 2,4,6-trinitrobenzenesulfonic acid (TNBS). Acute intestinal inflammation was induced in 40 male SD rats aged 4 weeks with 100 mg/kg TNBS, and then three dosages of SLBZS (0.5 g/kg, 1 g/kg, and 1.5 g/kg) were administered for eight days, respectively. Faecal microbiome composition was assessed by 16S rRNA high-throughput sequencing. The result indicated that SLBZS could reduce the diversity of gut microbiota and increased its abundance. At the genus level, the relative abundance of SCFAs producing bacteria including Prevotella and Oscillospira increased, while the relative abundance of opportunistic pathogens including Desulfovibrio and Bilophila decreased. Meanwhile, SLBZS could improve the lesions of colon and significantly reduce the level of MPO, increase the levels of SOD and CAT in rats' serum. These findings revealed that SLBZS was effective and possessed anticolitic activities in a rat model of UC by reducing macroscopical and microscopical colon injury, enhancing antioxidant capacity, and regulating gut microbiota.

Effects of Acute Diquat Poisoning on Liver Mitochondrial Apoptosis and Autophagy in Ducks.

Diquat (DQ) is an effective herbicide and is widely used in agriculture. Due to persistent and frequent applications, it can enter into aquatic ecosystem and induce toxic effects to exposed aquatic animals. The residues of DQ via food chain accumulate in different tissues of exposed animals including humans and cause adverse toxic effects. Therefore, it is crucial and important to understand the mechanisms of toxic effects of DQ in exposed animals. We used ducks as test specimens to know the effects of acute DQ poisoning on mechanisms of apoptosis and autophagy in liver tissues. Results on comparison of various indexes of visceral organs including histopathological changes, apoptosis, autophagy-related genes, and protein expression indicated the adverse effects of DQ on the liver. The results of our experimental trial showed that DQ induces non-significant toxic effects on pro-apoptotic factors like BAX, BAK1, TNF-α, caspase series, and p53. The results revealed that anti-apoptotic gene Parkin was significantly upregulated, while an upward trend was also observed for Bcl2, suggesting that involvement of the anti-apoptotic factors in ducklings plays an important role in DQ poisoning. Results showed that DQ significantly increased the protein expression level of the autophagy factor Beclin 1 in the liver. Results on key autophagy factors like LC3A, LC3B, and p62 showed an upward trend at gene level, while the protein expression level of both LC3B and p62 reduced that might be associated with process of translation affected by the pro-apoptotic components such as apoptotic protease that inhibits the occurrence of autophagy while initiating cell apoptosis. The above results indicate that DQ can induce cell autophagy and apoptosis and the exposed organism may resist the toxic effects of DQ by increasing anti-apoptotic factors.

Effect of paraquat on cytotoxicity involved in oxidative stress and inflammatory reaction: A review of mechanisms and ecological implications.

Paraquat (PQ) is a cheap and an effective herbicide, which is widely being used worldwide to remove weeds in cultivated crop fields. However, it can cause soil and water pollution, and pose serious harm to the environment and organisms. Several countries have started to limit or prohibit the use of PQ because of the increasing number of human deaths. Its toxicity can damage the organisms with a multi-target mechanism, which has not been fully understood yet. That is why it is hard to treat as well. The current research on PQ focuses on its targeted organ, the lungs, in which PQ mostly trigger pulmonary fibrosis. While there is a lack of systematic research, there are few studies published discussing its toxic effects at systematic level. This review summarizes the major damages caused by PQ in different organisms and partial mechanisms by which it causes these damages. For this purpose, we consulted several research articles that studied the toxicity of PQ in various tissues. We also listed some drugs that can be used to alleviate the toxicity of PQ. However, at present, the effectiveness of these drugs is still being explored in animal experiments and the study of their mechanism will also help in understanding the poisoning mechanism of PQ, which will ultimately lead to effective treatment in future.

Effects of Heat Stress on Gut Microbiome in Rats.

Gut microbiome, as the largest and most important micro-ecosystem, plays a critical role in health. The purpose of this study was to evaluate whether heat stress modulates the composition and diversity of the gut microbiome in rats. The heat stress model was prepared in rats with the heating temperature maintained at 35-38°C. Cecum contents were collected after heat stress for 3 h and days 1, 3 and 7. Total DNA was extracted for 16 S rRNA sequencing and analysis of intestinal microbiome composition and diversity. The study showed that the composition of the intestinal microbiome of heat stress group was changed. And the heat stress modulated key phylotypes of gut microbiota at the level of phylum and genus. In particular, the genus of Lactobacillus and Bacteroides were significantly reduced, whereas the Oscillospira and Clostridium were increased by heat stress. Meanwhile, the rats under the heat stress encountered the change in carbohydrate metabolism, amino acid metabolism, and membrane transport to defense against stress. Taken together, the composition and structure of gut microbiome were affected by heat stress and some key phylotypes were also significantly altered. We conclude that the heat stress could impact multiple biological functions, via altering the gut microbiome.

Retracted: Inhibition of Nasopharyngeal Carcinoma by Beta-Lapachone Occurs by Targeting the Mammalian Target of Rapamycin (mTOR)/PI3K/AKT Pathway, Reactive Oxygen Species (ROS) Production, and Autophagy Induction.

Approval for publication of this manuscript was not obtained by all the authors, which is in breach of this journal's editorial guidelines. Reference: Yongqing Han, Dayou Shi, Jingao Li: Inhibition of Nasopharyngeal Carcinoma by Beta-Lapachone Occurs by Targeting the Mammalian Target of Rapamycin (mTOR)/PI3K/AKT Pathway, Reactive Oxygen Species (ROS) Production, and Autophagy Induction.  Med Sci Monit 2019; 25:8995-9002. 10.12659/MSM.915463.

Inhibition of Nasopharyngeal Carcinoma by Beta-Lapachone Occurs by Targeting the Mammalian Target of Rapamycin (mTOR)/PI3K/AKT Pathway, Reactive Oxygen Species (ROS) Production, and Autophagy Induction.

BACKGROUND Beta-lapachone has been shown to exhibit potent anti-cancer effects against various cell lines. In the present study, we examined the anti-cancer effects of beta-lapachone, a quinone, against human HNE1 nasopharyngeal carcinoma cells, and also assessed its effects on cellular migration and invasion, autophagy, mTOR/PI3K/AKT signalling pathway, and ROS production. MATERIAL AND METHODS CCK-8 cell counting assay was used to assess cell viability effects after lapachone treatment. Its effects on the mTOR/PI3K/AKT biochemical pathway were examined by Western blot analysis. Transmission electron microscopy was used to study autophagy induced by beta-lapachone. Effects on cell invasion and cell migration were evaluated by Transwell method. RESULTS The results revealed that beta-lapachone suppresses the proliferation of HNE1 cells, with an IC50 of 30 µM. These growth-inhibitory effects of beta-lapachone were found to be dose-dependent. The investigation of the effects of beta-lapachone on the mTOR/PI3KAKT signalling pathway showed that beta-lapachone blocked this pathway in a concentration-dependent manner. Beta-lapachone also inhibited the migration and invasion of HNE1 nasopharyngeal cancer cells, as shown by Transwell assay. The fluorescence microscopy analysis showed that beta-lapachone increased production of reactive oxygen species (ROS), which is also linked with a concentration-dependent decrease in mitochondrial membrane potential (MMP) levels. Electron microscopy analysis showed that beta-lapachone caused the development of the autophagosomes, and the frequency of the autophagosomes increased with increased dosage of beta-lapachone. The beta-lapachone-triggered autophagy was also associated with increased protein levels of LC3 II and decreased levels of p62. CONCLUSIONS The findings of this study suggest that beta-lapachone inhibits the growth of nasopharyngeal cancer cells by promoting autophagy, and it may be useful in cancer drug discovery paradigms.

Morphological characterization and molecular profiling of malignant pericardial effusion in patients with pulmonary adenocarcinoma.

CONTEXT: Malignant pericardial effusions (MPCEs) is a common complication observed in advanced pulmonary adenocarcinoma. In such cases, investigating molecular alterations can have significant therapeutic implication in determining anticancer drugs. AIM: The objective was to evaluate the significance of cell block technique in the diagnosis of MPCE and further investigate the morphological and molecular profiles of MPCE in patients with pulmonary adenocarcinoma. SETTING AND DESIGN: Cytopathological and molecular profiles of 19 MPCE cases in patients with pulmonary adenocarcinoma were retrospectively analyzed. The control group consisted of 14 malignant pleural effusion (MPE) cases in patients with pulmonary adenocarcinoma. MATERIALS AND METHODS: Anaplastic lymphoma kinase (ALK) and tyrosine-protein kinase Met (C-MET) expression was evaluated by fluorescence in situ hybridization (FISH). Epithelial growth factor receptor (EGFR) and K-Ras (KRAS) mutations were detected by ARMS real-time polymerase chain reaction (RT-PCR). STATISTICAL ANALYSIS USED: Associations between MPCE and MPE were analyzed using Fisher's exact test. RESULTS: MPCE was found to have micropapillary and solid pattern predominant with mucin secretion compared to acinar patterns, as seen in MPE. Seventeen MPCE cases (89.5%) and all MPE cases (100%) underwent molecular analysis. Mutations in EGFR and KRAS, ALK rearrangement, and C-MET amplification were observed in MPCE and MPE with statistical differences. Additionally, two MPCE cases demonstrated EGFR T790M mutation and multiple insertions at L858. CONCLUSIONS: MPCE shows micropapillary and solid cytological patterns predominant with mucin secretion. MPCE are suitable to analyze oncogenic mutations and to develop targeted therapy for patients with pulmonary adenocarcinoma. Further molecular investigations may reveal novel molecular alterations.

The immunoregulatory effect of sulfated Echinacea purpurea polysaccharide on chicken bone marrow-derived dendritic cells.

Echinacea purpurea polysaccharide (EPP) was obtained by modern separation technology and sulfated EPP (sEPP) was prepared by sulfation modification. The immunological effects of EPP and sEPP were compared on chicken bone marrow-derived dendritic cells (chBM-DCs). The results showed that the surface marker expression of CD11c and CD80 was increased after chBM-DCs were cultured with three dosage of sEPP, especially in sEPPM group. Three dosage of sEPP, EPPL and LPS could significantly enhance the effects of chBM-DCs on the proliferation of allogenic mixed lymphocytes. After chBM-DCs treatment with EPP or sEPP in vitro, the levels of IL-2 of sEPPH and EPPM groups were significantly higher than those of LPS group (P < 0.05). All sEPP and EPP groups could enhance the level of IFN-γ and down-regulated the level of IL-4 and IL-10. Results indicated that both sEPP and EPP had immunoregulatory effects on chBM-DCs, sEPP possessed better immunoregulatory effects as compared with EPP.

Impact of gut microbiota structure in heat-stressed broilers.

Gut microbiota** play important roles in the health and disease status of both humans and animals. Little is known about whether heat stress changes the composition of the gut microbiota in chicken. The aim of this study was to investigate the effects of heat stress on changes in caecal microbiota, including changes in growth performance as well as HSP70 and cortisol levels. Sixty 14-day-old female broilers were equally divided into 2 treatment groups with different housing temperatures for 28 D: a control group (C) at 24 to 26°C and a heat stress (HS) group at 34 to 38°C. The caecal contents of the broiler chicken were then extracted on days 1, 3, 7, 14, and 28. Genomic DNA was extracted and amplified based on the V3∼V4 hypervariable region of 16S rRNA high-throughput sequence analyses. The results showed that the average daily gain and average daily feed intake were significantly decreased and that the feed conversion ratio was increased by heat stress. The concentrations of HSP70 and cortisol in the serum were significantly increased. The composition of gut microbiota was influenced by heat stress** through beta diversity analysis and taxon-based analysis. In particular, at the phylum level the composition of Firmicutes, Tenericutes, and Proteobacteria in HS group was increased than that of C group, and Bacteroidetes and Cyanobacteria in HS group were reduced than that of C group. In addition, the composition of Anaeroplasma and Lactobacillus phyla in HS group were increased than that of C group, whereas the Bacteroides, Oscillospira, Faecalibacterium, and Dorea genera in HS group were decreased than that of C group. In conclusion, the gut microbiota in broilers were changed by heat stress. And the changes of the gut microbiota could provide the basis for further research on the heat stress.

Explore the potential effect of natural herbals to resist Newcastle Disease Virus.

In this study, we examined the dose-dependent effects of the formula on Newcastle disease virus (NDV). In in-vitro test, the formula within safety concentration scope and NDV were added into cultured chick embryo fibroblast in 3 modes, and the cellular A570 values were determined by MTT (3-(4, 5-dimethyithiazol-2-yl)-2, 5-diphenyltetrazolium bromide) method. In in-vivo test, we examined the expression of interferon-induced transmembrane protein 3 (IFITM3) and Interferons (IFNs) in NDV-infected chickens. The results showed that the highest virus inhibitory rates of the formula at optimal concentration group were the highest (15.625 mg/mL) in post-adding and simultaneous-adding drug and virus modes, whereas medium concentration (7.813 mg/mL) showed the highest virus inhibitory rates in pre-adding drug mode. In vivo, the formula significantly upregulated the expression of IFITM3 in NDV-infected chickens at 3-D post-infection. However, the levels of IFNs were significantly downregulated. On days 5 and 7 post-infection, the levels of IFNs quickly upregulated. Moreover, the formula can significantly upregulate the antibody to resist the NDV compared with model control group on days 5 and 7 post-infection. In animals treated with the formula, the survival rate was nearly 37% higher at 7 d post-infection. We also found that the formula had a significantly stronger effect than a single herb on upregulating the expression of IFITM3. It confirmed that the formula could significantly inhibit the infectivity of NDV to chick embryo fibroblast. Also, the formula could significantly upregulated IFITM3 expression and inhibited virus replication in NDV-infected chickens. During the early stage of infection, IFNs were consumed to stimulate IFITM3 to inhibit virus replication, whereas during later stages of the infection, the formula upregulated the levels of IFNs and their antibodies to maintain a high level of immunity.

Structural modulation of gut microbiota during alleviation of antibiotic-associated diarrhea with herbal formula.

The gut microbiome is hypothesized to play a critical role in gastrointestinal diseases, including antibiotic-associated diarrhea (AAD). To determine whether the traditional Chinese herbal formula of Shen Ling Bai Zhu San (SLBZS) modulates the composition of the gut microbiome during AAD treatment, an AAD diarrhea model was prepared in rats by gastric gavage with lincomycin for 7 successive days, followed by administration of SLBZS for one week. At all time points after the SLBZS treatment, the diarrhea rates were significantly or at least numerically lower than that of the untreated model group. Overall structural modulation of the gut microbiome occurred after SLBZS treatment, with reverting effects on the AAD-induced structural variations. At the genus level, the relative abundance of Sutterella was negatively correlated with SLBZS treatment and positively correlated with a lack of treatment, suggesting that Sutterella might be a pivotal phylotype associated with the improvement of AAD. The key phylotypes of the gut microbiome that responded to SLBZS indicated enrichment of beneficial bacteria, and particularly Bacteroides spp. These data therefore demonstrated that structural changes of the gut microbiome are induced by the Chinese herbal formula SLBZS. In conclusion, changes in the gut microbiome are associated with the diarrhea-controlling effect of SLBZS.