TM6SF1 suppresses the progression of lung adenocarcinoma and M2 macrophage polarization by inactivating the PI3K/AKT/mtor pathway.

Transmembrane 6 superfamily 1 (TM6SF1) is lowly expressed in lung adenocarcinoma (LUAD), but the function and mechanisms of TM6SF1 remain unclear. Thus, we attempt to explore the function of TM6SF1 and its underlying mechanisms in LUAD. qRT-PCR was used for detecting TM6SF1 mRNA expression. Immunohistochemistry staining was used for detecting the expression of MMP-2, TM6SF1, Ki67, MMP-9, and CD163 proteins. E-cadherin, p-PI3K, Vimentin, AKT, N-cadherin, PI3K, p-AKT, mTOR, p-mTOR, and marker proteins of M2 macrophages were evaluated using Western blot. CD206 protein expression was examined via immunofluorescence. The IL-10 concentration was measured via enzyme-linked immunosorbent assay (ELISA). Using CCK-8, colony formation and transwell assays, cell proliferation, migration, and invasion were assessed. A549 cells were injected into the mice's flank for establishing a mouse tumor model and into the tail vein for establishing the lung metastasis model. HE staining was performed to detect pathological changes in lung tissues. Decreased TM6SF1 expression was found in LUAD tissues and cells. TM6SF1 overexpression inhibited cell viability, proliferation, invasion, migration, EMT, and polarization of M2 macrophages in LUAD cells, along with tumor growth and metastasis in xenograft mice. Bioinformatics analysis demonstrated that TM6SF1 was correlated with the tumor microenvironment. TM6SF1 overexpression reduced expression levels of p-mTOR, p-PI3K, p-AKT, mTOR, and AKT. TM6SF1-caused inhibition of proliferation, migration, invasion and EMT, as M2 macrophage polarization was reversed by the PI3K activator in LUAD cells. TM6SF1 inactivated the PI3K/AKT/mTOR pathway to suppress LUAD malignancy and polarization of M2 macrophages, providing insight for developing new LUAD treatments.

Up-regulation of inflammatory, oxidative stress, and apoptotic mediators via inflammatory, oxidative stress, and apoptosis-associated pathways in bovine endometritis.

Endometritis is the inflammation of the endothelial lining of the uterine lumen and is multifactorial in etiology. Escherichia (E.) coli is a Gram-negative bacteria, generally considered as a primary causative agent for bovine endometritis. Bovine endometritis is characterized by the activation of Toll-like receptors (TLRs) by E. coli, which in turn triggers inflammation, oxidative stress, and apoptosis. The objective of this study was to investigate the gene expression of inflammatory, oxidative stress, and apoptotic markers related to endometritis in the uteri of cows. Twenty uterine tissues were collected from the abattoir. Histologically, congestion, edema, hyperemia, and hemorrhagic lesions with massive infiltration of neutrophil and cell necrosis were detected markedly (P < 0.05) in infected uterine samples. Additionally, we identify E. coli using the ybbW gene (177 base pairs; E. coli-specific gene) from infected uterine samples. Moreover, qPCR and western blot results indicated that TLR2, TLR4, proinflammatory mediators, and apoptosis-mediated genes upregulated except Bcl-2, which is antiapoptotic, and there were downregulations of oxidative stress-related genes in the infected uterine tissue. The results of our study suggested that different gene expression regimes related to the immune system reflex were activated in infected uteri. This research gives a novel understanding of active immunological response in bovine endometritis.

Effects of supplemental Glycyrrhiza polysaccharide on growth performance and intestinal health in weaned piglets.

In this study, we investigated the effects of supplemental Glycyrrhiza polysaccharide (GCP) on growth performance and intestinal health of weaned piglets. Ninety piglets weaned at 28 days of age were randomly allocated to three groups with five replicates per treatment. Piglets were fed the following diets for 28 days: (1) CON (control group), basal diet; (2) G500, CON + 500 mg/kg GCP; (3) G1000, CON + 1000 mg/kg GCP. The results showed that supplementation with 1000 mg/kg GCP increased the average daily gain (ADG) and decreased the feed-to-gain ratio (F/G) (P < 0.05). Serum diamine oxidase (DAO) and D-lactic acid (DL-A) levels were lower in the G1000 group (P < 0.05). Dietary GCP 1000 mg/kg improved mucosal trypsin activity in the duodenum, jejunum and ileum and increased lipase and amylase activity in the jejunum (P < 0.05). Moreover, in the G1000 group, ZO-1, claudin 1 and occludin levels were increased in the jejunum mucosa, whereas interleukin-1ß (IL-1ß) and IL-6 levels were decreased (P < 0.05). The 16S rRNA gene analysis indicated that dietary 1000 mg/kg GCP altered the jejunal microbial community, with increased relative abundances of beneficial bacteria. In conclusion, dietary GCP 1000 mg/kg can improve growth performance, digestive enzyme activity, intestinal immunity, barrier function and microbial community in weaned piglets.

Effects of dietary Glycyrrhiza polysaccharide on growth, serum biochemistry, immunity, and egg laying in quail.

This study was conducted to evaluate the effects of dietary supplementation with Glycyrrhiza polysaccharide (GCP) on growth performance, serum biochemistry, immunity, and egg laying in female quail. 300 1-day-old female quail were sorted into four dietary treatments with five replicate cages of 15 birds each. The basic diet in the four treatment groups was supplemented with 0, 500, 1000 and 1500 mg/kg GCP, and the experiment continued for 80 days. Results showed that dietary supplementation with GCP significantly (p < 0.05) increased average daily gain in a dose-dependent fashion, and decreased (p < 0.05) the feed-to-gain ratio and mortality. The relative weights of the thymus and bursa of Fabricius increased (p < 0.05) linearly with increasing dose of GCP from 0 to 1500 mg/kg on day 20. GCP birds showed higher serum levels of protein, glucose, immunoglobulin A and immunoglobulin M, but lower serum triglycerides (p < 0.05) on day 50. GCP increased (p < 0.05) average laying rate and average egg weight linearly from days 60 to 80, whereas feed to egg ratio was decreased (p < 0.05). Taken together, these results revealed that GCP could improve growth performance, serum biochemistry, immunity, and egg laying in female quail. Therefore, GCP may be a potential replacement for antibiotic growth promoters in poultry.

Effects of dietary Glycyrrhiza polysaccharide supplementation on growth performance, intestinal antioxidants, immunity and microbiota in weaned piglets.

The aim of this study was to evaluate the effects of dietary Glycyrrhiza polysaccharide (GCP) supplementation on growth performance, intestinal antioxidants, immunity and microbiota in weaned piglets. One hundred and twenty 28-day-old weaned piglets were randomly assigned into five groups (four replicates per group) and fed a basal diet with GCP at 0, 500, 1000, 2000 and 4000 mg/kg for four weeks, respectively. Results showed that 1000 mg/kg GCP improved piglets' ADG and ADFI and reduced FCR (p < .05). Thus, the 0 and 1000 mg/kg GCP dose were selected for subsequent experiments. We found that 1000 mg/GCP increased SOD and T-AOC and decreased MDA in the jejunal mucosa (p < .05). Dietary 1000 mg/kg GCP also resulted in high levels of sIgA, IL-10 and TGF-ß, whereas IL-2 dropped dramatically (p < .05). The relative expression levels of ZO-1, CLDN, OCLDN, TLR-4, IL-10, TGF-ß, Nrf-2, SOD1 and CAT increased in the jejunal mucosa, whereas INF-γ decreased (p < .05). 1000 mg/kg GCP treatment altered the diversity and community composition of cecal microbiota in pigs, with increasing relative abundance of Bacteroidota and Lactobacillus at phylum and genus levels (p < .05), respectively. The results suggested that dietary 1000 mg/kg GCP could improve growth performance and intestinal health of weaned piglets.

Transcriptomic profiling of lipopolysaccharide-challenged bovine mammary epithelial cells treated with forsythoside A.

Mastitis is usually caused by a variety of pathogenic bacteria that seriously impact the health and milk-production ability of dairy cows, with consequent, economically detrimental effects on the dairy industry. Forsythoside A (FTA), isolated from the fruit and leaves of Forsythia suspensa (Thunb.) Vahl (Oleaceae), has been reported to have significant antioxidant, anti-inflammatory, and antibacterial effects. However, it is not clear whether FTA exerts a protective effect against lipopolysaccharide (LPS)-induced bovine mastitis and its potential gene signature. In this study, high-throughput sequencing technology was performed to analyze the differences between the mRNA and enrichment pathway of bovine mammary epithelial cells of the control, LPS, and LPS + FTA groups. The results showed that there were 139 differentially expressed genes (DEGs) (p-value < 0.05, |log2FoldChange| > 1, FPKM > 1) in the LPS group compared with the control group, including 121 up-regulated genes and 18 down-regulated genes, which were mainly enriched in the cellular response to lipopolysaccharide, cytokine activity, protein binding, and IL-17 signaling pathway based on Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis, respectively. Compared with the control group and LPS + FTA group, there were 349 DEGs, including 322 up-regulated genes and 27 down-regulated genes. They were mainly enriched in protein localization to organelles, centrosomes, binding, and the IL-17 signaling pathway, based on GO and KEGG analysis. Compared to the LPS group, the LPS + FTA group had 272 DEGs, including 259 up-regulated genes and 13 down-regulated genes, which were mainly enriched in RNA processing, IL-6 receptor binding, and the lysosome pathway, based on GO and KEGG analyses. It can be seen that LPS stimulation induced the expression of inflammation-related genes, IL-17 and IL-6, whereas FTA treatment promoted the expression of the spliceosome-, lysosome-, and oxidative stress-related genes HSP70, HSPA8, and PARP2. The study utilized RNA-sequencing analysis of FTA against LPS-challenged bovine mammary epithelial cells to explore key mRNA findings that may be strongly associated with inflammation and oxidative stress, and provides a theoretical reference for further elucidation of molecular mechanisms of bovine mastitis and therapeutic effects of FTA against bovine mastitis.

Leg disorders in broiler chickens: a review of current knowledge.

Ensuring improved leg health is an important prerequisite for broilers to achieve optimal production performance and welfare status. Broiler leg disease is characterized by leg muscle weakness, leg bone deformation, joint cysts, arthritis, femoral head necrosis, and other symptoms that result in lameness or paralysis. These conditions significantly affect movement, feeding and broiler growth performance. Nowadays, the high incidence of leg abnormalities in broiler chickens has become an important issue that hampers the development of broiler farming. Therefore, it is imperative to prevent leg diseases and improve the health of broiler legs. This review mainly discusses the current prevalence of broiler leg diseases and describes the risk factors, diagnosis, and prevention of leg diseases to provide a scientific basis for addressing broiler leg health problems.

A Perifacial EMG Acquisition System for Facial-Muscle-Movement Recognition.

This paper proposes a portable wireless transmission system for the multi-channel acquisition of surface electromyography (EMG) signals. Because EMG signals have great application value in psychotherapy and human-computer interaction, this system is designed to acquire reliable, real-time facial-muscle-movement signals. Electrodes placed on the surface of a facial-muscle source can inhibit facial-muscle movement due to weight, size, etc., and we propose to solve this problem by placing the electrodes at the periphery of the face to acquire the signals. The multi-channel approach allows this system to detect muscle activity in 16 regions simultaneously. Wireless transmission (Wi-Fi) technology is employed to increase the flexibility of portable applications. The sampling rate is 1 KHz and the resolution is 24 bit. To verify the reliability and practicality of this system, we carried out a comparison with a commercial device and achieved a correlation coefficient of more than 70% on the comparison metrics. Next, to test the system's utility, we placed 16 electrodes around the face for the recognition of five facial movements. Three classifiers, random forest, support vector machine (SVM) and backpropagation neural network (BPNN), were used for the recognition of the five facial movements, in which random forest proved to be practical by achieving a classification accuracy of 91.79%. It is also demonstrated that electrodes placed around the face can still achieve good recognition of facial movements, making the landing of wearable EMG signal-acquisition devices more feasible.

Effects of dietary Glycyrrhiza polysaccharide on growth performance, blood parameters and immunity in weaned piglets.

The purpose of this study was to evaluate the effects of dietary Glycyrrhiza polysaccharide (GCP) on growth performance, blood parameters and immunity in weaned piglets. A total of 240 (10.33 ± 0.62 kg body weight) 35-day-old (Duroc × Landrace × White) weaned piglets were randomly assigned to four dietary treatments, with six replicate pens per treatment and 10 piglets per pen (five males and five females). The dietary treatments continued for 21 days and comprised a basal diet supplemented with 0 (control group), 500, 1000 and 2000 mg/kg GCP. The results showed that the inclusion of 1000 and 2000 mg/kg GCP increased the average daily gain and decreased the feed conversion rate compared with the control group (p < 0.05). The piglets treated with 500 and 1000 mg/kg GCP had a lower diarrhoeal incidence than the control group (p < 0.05). Moreover, supplementation with 1000 mg/kg GCP increased the counts of white blood cells, neutrophils, red blood cells, and platelets, and elevated alkaline phosphatase, total protein, globulin, glucose, triglyceride, immunoglobulin A, immunoglobulin G, and total antioxidant capacity levels (p < 0.05), and decreased malondialdehyde content compare with the control group (p < 0.05). In addition, relative to the control group, piglets fed 500 and 1000 mg/kg GCP had significantly lower expression of interleukin-6 mRNA in spleen (p < 0.05). Our results indicate that dietary supplementation with GCP can improve growth performance, blood parameters and immunity in weaned piglets. Our study suggests that adding 1000 mg/kg GCP to the diet had the most beneficial effect.

Dual-ATME: Dual-Branch Attention Network for Micro-Expression Recognition.

Micro-expression recognition (MER) is challenging due to the difficulty of capturing the instantaneous and subtle motion changes of micro-expressions (MEs). Early works based on hand-crafted features extracted from prior knowledge showed some promising results, but have recently been replaced by deep learning methods based on the attention mechanism. However, with limited ME sample sizes, features extracted by these methods lack discriminative ME representations, in yet-to-be improved MER performance. This paper proposes the Dual-branch Attention Network (Dual-ATME) for MER to address the problem of ineffective single-scale features representing MEs. Specifically, Dual-ATME consists of two components: Hand-crafted Attention Region Selection (HARS) and Automated Attention Region Selection (AARS). HARS uses prior knowledge to manually extract features from regions of interest (ROIs). Meanwhile, AARS is based on attention mechanisms and extracts hidden information from data automatically. Finally, through similarity comparison and feature fusion, the dual-scale features could be used to learn ME representations effectively. Experiments on spontaneous ME datasets (including CASME II, SAMM, SMIC) and their composite dataset, MEGC2019-CD, showed that Dual-ATME achieves better, or more competitive, performance than the state-of-the-art MER methods.

Total flavonoids of Rhizoma Drynariae protect hepatocytes against aflatoxin B1-induced oxidative stress and apoptosis in broiler chickens.

Aflatoxin B1 (AFB1) is a common mycotoxin in food and in the environment that lead to multi-organ injury in humans and animals. The objective of this study was to evaluate the detoxification properties of dietary total flavonoids of Rhizoma drynariae (TFRD), a Chinese herbal, on aflatoxin B1 (AFB1)-induced hepatic oxidative damage and apoptosis of liver of broiler chickens. A total of 160 healthy specific pathogen free (SPF) 21-day-old broilers were randomly allocated to 4 groups, including the CON group (basal diet), TFRD group (basal diet with 125 mg/kg TFRD), AFB1 group (100 µg/kg body weight), and AFB1 (100 µg/kg body weight) + TFRD (basal diet with 125 mg/kg TFRD) group. The exposure of AFB1 continued for seven days. The results showed that TFRD treatment alleviated the abnormal changes of growth performance and liver morphology, reduced serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels. Moreover, TFRD promoted the antioxidant capacity of serum, increased the activities of total superoxide dismutase (T-SOD), oxidized glutathione (GSSG) and glutathione (GSH) (p < 0.05), while decreased MDA contents (p > 0.05). Meanwhile, supplementation of TFRD significantly increased the expression of antioxidant-related genes (SOD, CAT, GST, and GPX1) in liver (p < 0.05). Furthermore, we found that AFB1 was involved in the regulation of PI3K/AKT signaling pathway, leading to hepatocyte apoptosis. At the same time, TFRD treatment inhibited AFB1-induced apoptosis and significantly changed mRNA expression of apoptosis-related genes, including PI3K, AKT, Bax, and Bcl-2 (p < 0.05). The results indicated that TFRD could alleviate AFB1-induced liver injury in broiler chickens.

Aflatoxin B1 causes oxidative stress and apoptosis in sheep testes associated with disrupting rumen microbiota.

Aflatoxin B1 (AFB1) is an unavoidable environmental pollutant commonly found in feed and foodstuffs. It is the most toxic one of all the aflatoxins, which can cause severe impairment to testicular development and function. Yet, the underlying mechanisms of reproductive toxicity in rams sheep remain inconclusive. The study was designed to explore the effects of AFB1 on sheep testes through rumen-microbiota, oxidative stress and apoptosis. Six-month-old male Dorper rams (n = 6) were orally administrated with 1.0 mg/kg AFB1 (dissolved in 20 mL 4% ethanol) 24 h before the experiment. At the same time, rams in the control group (n = 6) were intragastrically administrated with 20 mL 4% ethanol. It was observed that acute AFB1 poisoning had significant (p < 0.05) toxin residue in the testis and could cause testicular histopathological damage. AFB1 stimulated the secretion of plasma testosterone level through regulating testosterone synthesis-related genes (StAR, 3ß-HSD, CYP11A1, and CYP17A1), which are accompanied by the increase of oxidative stress and testicular apoptosis that had a close relationship with the regulation of testosterone secretion. Interestingly, we observed rumen dysbacteriosis and decreased the abundances of Prevotella, Succiniclasticum, CF231, Ruminococcus, and Pseudobutyrivibrio in AFB1-exposed sheep, which were negatively correlated to the testosterone synthesis-related gene levels. Taken together, our findings indicated that AFB1 induced testicular damage and testicular dysfunction, which is related to testicular oxidative stress and apoptosis involved in rumen dysbacteriosis in sheep.

Icariin Alleviates Escherichia coli Lipopolysaccharide-Mediated Endometritis in Mice by Inhibiting Inflammation and Oxidative Stress.

Icariin (ICA) is a naturally occurring phytochemical agent primarily extracted from Epimedium Brevicornum Maxim (Family Berberidaceae) with a broad spectrum of bioactivities. Endometritis is a uterine disease that causes enormous losses in the dairy industry worldwide. In this study, anti-inflammatory and anti-oxidant properties of ICA were investigated against lipopolysaccharide (LPS)-induced endometritis in mice to investigate possible underlying molecular mechanisms. Sixty heathy female Kunming mice were randomly assigned to four groups (n = 15), namely control, LPS, LPS + ICA, and ICA groups. The endometritis was induced by intrauterine infusion of 50 µL of LPS (1 mg/mL). After 24 h of onset of LPS-induced endometritis, ICA groups were injected thrice by ICA intraperitoneally six hours apart. Histopathological examination, enzyme linked immunosorbent assay (ELISA), real time quantitative polymerase chain reaction (RT-qPCR), western blotting, and immunohistochemistry were used in this study. Histological alterations revealed that ICA markedly mitigated uterine tissue injury caused by LPS. The results showed that the ICA inhibited the production of pro-inflammatory cytokines (IL-1ß, IL-6, and TNF-α) and boosted the production of anti-inflammatory cytokines (IL-10). Additionally, ICA modulated the expression of malondialdehyde (MDA), reactive oxygen species (ROS), superoxide dismutase 1 (SOD1), catalase (CAT), and glutathione peroxidase 1 (Gpx1) induced by LPS. The administration of ICA significantly (p < 0.05) improved the mRNA and protein expression of Toll-like receptor (TLR) 4. The western blotting and ELISA finding revealed that the ICA repressed LPS-triggered NF-κB pathway activation. Moreover, ICA improved the antioxidant defense system via activation of the Nrf2 pathway. The results revealed that ICA up-regulated the mRNA and protein expression of Nuclear erythroid-2-related factor (Nrf2), NAD(P)H: quinone oxidoreductase 1 (NQO1), heme oxygenase-1 (HO-1), and glutamate-cysteine ligase catalytic subunit (GCLC) under LPS exposure. Conclusively, our findings strongly suggested that ICA protects endometritis caused by LPS by suppressing TLR4-associated NF-κB and Nrf2 pathways. Altogether, these innovative findings may pave the way for future studies into the therapeutic application of ICA to protect humans and animals against endometritis.

Aflatoxin B1 alters meat quality associated with oxidative stress, inflammation, and gut-microbiota in sheep.

Aflatoxin B1 (AFB1) is an unavoidable contaminant in animal feed and agricultural products. AFB1 has been found to impair the liver and kidney function of sheep. However, few data are available, which explain the toxic damage of AFB1 exposure on meat quality. In the study, male Dorper RAMS sheep (6-month-old) were orally administrated with AFB1 at the dose of 1 mg/kg body weight once. The body temperature, serum biochemistry, meat quality-related parameters, oxidation indicators in meat and serum, the mRNA expression of pro-inflammatory cytokines and anti-inflammatory, and microbiota composition of feces were measured 24 h after AFB1 exposure. The results showed that the body temperature was slightly increased, the mental state of mutton sheep was suppressed, and biochemical indicators were significantly changed after AFB1 exposure. AFB1 impaired mutton quality reflected by the structure of muscle fibers was changed, and increased muscle drip loss and lightness (L*), and decreased muscle redness (a*). Moreover, we found that AFB1 caused changes in the oxidative stress indicators T-SOD, T-AOC, MDA, GSH level, and GSH/GSSG ratio, and inflammation damage of mutton reflected by increasing pro-inflammatory TNF-α and reducing anti-inflammatory IL-10 mRNA levels, disrupts the secretion of inflammatory factors, and changed the composition of gut microbiota reflected by significantly increased Firmicutes/Bacteroidetes ratio and decreased the abundances of Butyrivibrio, which are related to the quality of the mutton. In summary, gut microbiota participates in AFB1 to damage mutton quality, which may be co-mediated by oxidative stress, inflammation, and gut microbiota.

The fungicide thiram perturbs gut microbiota community and causes lipid metabolism disorder in chickens.

Fungicide thiram, a representative dithiocarbamate pesticide can cause potential health hazards to humans and animal health due to the residues in various agricultural products. However, the effects of thiram on lipid metabolism by perturbing gut microbiota of chickens are not clear. Our study was aimed to explore the protective of polysaccharide extracted from Morinda officinalis (MOP) on acute thiram-exposed chickens, and to analyze the association between alteration of gut microbiota and lipid metabolism. Three hundred chicks are fed with a normal diet, thiram-treated diet (100 mg/kg), and a thiram-treated diet supplemented with 250, 500, or 1000 mg/kg MOP was used in this study, respectively. The results showed that thiram exposure prominently elevated liver index, changed liver function by histopathological examination and serum biochemistry diagnoses, and increased blood lipid parameters. Meanwhile, the expression level of some key genes in hepatic lipid metabolism dysregulated significantly in the thiram-exposed chickens. Furthermore, 16S rRNA gene sequencing indicated that thiram exposure can significantly alter the richness, diversity, and composition of the broiler fecal microbiota, and the relative abundance of Firmicutes and Proteobacteria was also affected at the phylum level. In addition, some microbial populations including Lactobacillus, Ruminococcus, Oscillospira, Blautia, and Butyricicoccus significantly decreased at the genus level, whereas the Klebsiella was opposite. Correlation analysis further revealed a significant association between microorganisms and lipid metabolism-related parameters. Optimistically, 500 mg/kg MOP can alleviate the damage of thiram in the gut and liver. Together, these data suggest that thiram exposure causes the imbalance of the gut microbiota and hepatic lipid metabolism disorder in chickens.

Chlorogenic Acid Alleviates Thiram-Induced Tibial Dyschondroplasia by Modulating Caspases, BECN1 Expression and ECM Degradation.

Chlorogenic acid (CGA) is a widely applied traditional Chinese medicine ingredient which can be used for the treatment of osteoporosis. In this experiment, we investigated the potential therapeutic effect of chlorogenic acid on thiram-induced tibial dyschondroplasia (TD) and explored the underlying mechanisms that have been rarely mentioned by others yet. Performance indicator analysis and tibial parameter analysis showed that CGA exhibited a definite positive effect on thiram-induced TD chickens. In order to further explore the mechanisms underlying the positive actions of CGA, apoptotic, autophagic genes and MMPs involved in matrix mineralization of growth plate were evaluated in this study. The results showed that CGA decreased the expression of pro-apoptotic genes caspases-3 and caspases-9, leading to the reduction of apoptotic cells accumulated in growth plate. In addition, CGA also increased the level of BECN1, an important gene involved in autophagy, which benefits the survival of abnormal cells. Furthermore, CGA also increased the expression of MMP-9, MMP-10, and MMP-13, which can directly affect the ossification of bones. Altogether, these results demonstrate that CGA possesses a positive therapeutic effect on thiram-induced TD via modulating the expression of caspases and BECN1 and regulating the degradation of ECM (extracellular matrix).

Comparative analysis of gut microbial community in healthy and tibial dyschondroplasia affected chickens by high throughput sequencing.

Tibial dyschondroplasia (TD) is a common skeletal problem of avian species. The cause and etiology of this disorder is still not clear. The intestinal bacterial community is a complex ecosystem and plays very important role in healthy life of human and animals. Many researchers have made the connection between diseases and alteration of gut microbiota. However, little is know about the role of gut microbiota in TD. The objective of this study was to explore the diversity and composition of small and large intestinal bacterial communities of TD chickens. The intestinal contests were collected from healthy and TD chickens at day 18 of the trail. The V3-V4 region of 16S rDNA was amplified and sequenced by high-throughput sequencing. Our results indicated that healthy chickens had higher abundance and diversity of gut microbiota than TD chickens but there was no significant difference (P > 0.05) among the gut microbiota of small and large intestine. However, the composition of bacterial communities were significantly different (P < 0.05) between healthy and TD chickens. The occurrence of gut pathogens associated with immunity and inflammation were much higher in the intestinal contests of TD chickens as compared to healthy ones. In summary, our results showed an interesting difference of gut microbiota between healthy and TD chickens, which provided a new thought about the pathogenesis of TD.

Clinical efficiency and safety of Hsp90 inhibitor Novobiocin in avian tibial dyschondroplasia.

Tibial dyschondroplasia (TD) is a bone defect of broilers and other poultry birds that disturbs growth plate and it causes lameness. Previously we evaluated differential expression of multiple genes involved in growth plate angiogenesis and reported the safety and efficacious of medicinal plant root extracted for controlling TD. In this study, clinical and protective effect of an antibiotic Novobiocin (Hsp90 inhibitor) and expression of Hsp90 and proteoglycan aggrecan was examined. The chicks were divided into three groups; Control, thiram-induced TD, and Novobiocin injected TD. After the induction of TD, the Novobiocin was administered through intraperitoneal route to TD-affected birds until the end of the experiment. The expressions and localization of Hsp90 were evaluated by qRT-PCR, immunohistochemistry (IHC) and western blot, respectively. Morphological, histological examinations, and serum biomarker levels were evaluated to assess specificity and protective effects of Novobiocin. The results showed that TD causing retarded growth, enlarged growth plate, distended chondrocytes, irregular columns of cells, decreased antioxidant capacity, reduced protein levels of proteoglycan aggrecan, and upregulated in Hsp90 expression (p < 0.05) in dyschondroplastic birds as compared with control. Novobiocin treatment restored growth plate morphology, reducing width, stimulated chondrocyte differentiation, sprouting blood vessels, corrected oxidative imbalance, decreased Hsp90 expressions and increased aggrecan level. Novobiocin treatment controlled lameness and improved growth in broiler chicken induced by thiram. In conclusion, the accumulation of the cartilage and up-regulated Hsp90 are associated with TD pathogenesis and irregular chondrocyte morphology in TD is along with reduced aggrecan levels in the growth plate. Our results indicate that Novobiocin treatment has potential to reduce TD by controlling the expression of Hsp90 in addition to improve growth and hepatic toxicity in broiler chicken.

Experimental mouse lethality of Escherichia coli strains isolated from free ranging Tibetan yaks.

The present study has examined the virulence potential of Escherichia coli isolates harboring at least one virulence gene (associated with ExPEC or InPEC pathotype and belonging to different phylogenetic groups: A, B1, B2 or D), isolated from free ranging Tibetan yak feces. The E. coli isolates (n = 87) were characterized for different serogroups and a mouse model of subcutaneous-infection was used to envisage the virulence within these E. coli strains. Of the 87 E. coli isolates examined, 23% of the E. coli isolates caused lethal infections in a mouse model of subcutaneous infection and were classified as killer. Moreover, the majority of the killer strains belonged to phylogroup A (65%) and serogroup O60 or O101 (35%). Phylogroup B1, serogroups O60 and O101 were statistically associated with the killer status (P < 0.05). However, positive associations (OR >1) were observed between the killer status isolates and all other bacterial virulence traits. This study comprises the first report on the virulence potential of E. coli strains isolated from free-ranging Tibetan yaks feces. Our findings suggest that pathogenic E. coli of free ranging yaks is highly worrisome, as these feces are used as manures by farmers and therewith pose a health risk to humans upon exposure.

Evidence of JEV in Culex tritaeniorhynchus and pigs from high altitude regions of Tibet, China.

BACKGROUND & OBJECTIVES: Culex tritaeniorhynchus is the primary vector of Japanese encephalitis virus (JEV) which is a major threat to animals and humans health. This virus has been reported earlier from low altitude regions of Tibet, in mosquitoes, Tibetan pigs and local Tibetans, but no reports are available for the probable propagation of JE to high altitude regions (3100 m) of Tibet. This study aimed to find the evidence of JEV in Cx. tritaeniorhynchus and pigs from high altitude regions of Tibet, China. METHODS: In total, 102 porcine serum samples and eight pools of Cx. tritaeniorhynchus were subjected to real-time PCR (RT-PCR) for the amplification of a fragment (~ 420 bp) of the NS1 gene. The resultant amplicons of the genes were subsequently analyzed and sequenced. RESULTS: Overall seroprevalence of JE in Tibetan pigs was 6.86%, while five pools of Cx. tritaeniorhynchus were found positive for JEV. Results showed genotype III as the most prevalent (100%) among JEV positive isolates. Furthermore, phylogenetic analysis of the JEV positive strains showed 98.8-99% similarity to four reference strains from China (JEV-Hubei, Whe, HYZ and CQ11-66). INTERPRETATION & CONCLUSION: The present study revealed that JEV has extended its geographic range to high altitude regions of Tibet. The factors like increased tourism and transportation might play key role in the transmission of JE that comprises a potential health risk for humans and animals.