BRD9-mediated control of the TGF-ß/Activin/Nodal pathway regulates self-renewal and differentiation of human embryonic stem cells and progression of cancer cells.

Bromodomain-containing protein 9 (BRD9) is a specific subunit of the non-canonical SWI/SNF (ncBAF) chromatin-remodeling complex, whose function in human embryonic stem cells (hESCs) remains unclear. Here, we demonstrate that impaired BRD9 function reduces the self-renewal capacity of hESCs and alters their differentiation potential. Specifically, BRD9 depletion inhibits meso-endoderm differentiation while promoting neural ectoderm differentiation. Notably, supplementation of NODAL, TGF-ß, Activin A or WNT3A rescues the differentiation defects caused by BRD9 loss. Mechanistically, BRD9 forms a complex with BRD4, SMAD2/3, ß-CATENIN and P300, which regulates the expression of pluripotency genes and the activity of TGF-ß/Nodal/Activin and Wnt signaling pathways. This is achieved by regulating the deposition of H3K27ac on associated genes, thus maintaining and directing hESC differentiation. BRD9-mediated regulation of the TGF-ß/Activin/Nodal pathway is also demonstrated in the development of pancreatic and breast cancer cells. In summary, our study highlights the crucial role of BRD9 in the regulation of hESC self-renewal and differentiation, as well as its participation in the progression of pancreatic and breast cancers.

Fermented bile acids improved growth performance and intestinal health by altering metabolic profiles and intestinal microbiome in Micropterus salmoides.

A type of fermented bile acids (FBAs) has been produced through a biological method, and its effects on growth performance, metabolism, and intestinal microbiota in largemouth bass were investigated. The results demonstrated that incorporating 0.03 %-0.05 % FBAs diet could improve the final weight, weight gain and specific growth rate, and decrease the feed conversion ratio. Dietary FBAs did not significantly affect the levels of high-density lipoprotein, low-density lipoprotein, and triglycerides, but decreased the activities of α-amylase in most groups. Adding FBAs to the diet significantly increased the integrity of the microscopic structure of the intestine, thickened the muscular layer of the intestine, and notably enhanced its intestinal barrier function. The addition of FBAs to the diet increased the diversity of the gut microbiota in largemouth bass. At the phylum level, there was an increase in the abundance of Proteobacteria, Firmicutes, Tenericutes and Cyanobacteria and a significant decrease in Actinobacteria and Bacteroidetes. At the genus level, the relative abundance of beneficial bacteria Mycoplasma in the GN6 group and Coprococcus in the GN4 group significantly increased, while the pathogenic Enhydrobacter was inhibited. Meanwhile, the highest levels of AKP and ACP were observed in the groups treated with 0.03 % FBAs, while the highest levels of TNF-α and IL-10 were detected in the group treated with 0.04 % FBAs. Additionally, the highest levels of IL-1ß, IL-8T, GF-ß, IGF-1, and IFN-γ were noted in the group treated with 0.06 % FBAs. These results suggested that dietary FBAs improved growth performance and intestinal wall health by altering lipid metabolic profiles and intestinal microbiota in largemouth bass.

Publisher Correction: Chromatin analysis in human early development reveals epigenetic transition during ZGA.

In this Letter, the 'Open chromatin' label in Fig. 4a should have been centred above the first three columns, and the black horizontal line underneath the label should have been removed. In addition, there should have been a vertical black line between the last two sets of panels for consistency. Minor changes have also been made to Fig. 1 and to the legend of Fig. 3. These errrors have been corrected online, and see Supplementary Information to the accompanying Amendment for the original Fig. 4.

Chromatin analysis in human early development reveals epigenetic transition during ZGA.

Upon fertilization, drastic chromatin reorganization occurs during preimplantation development 1 . However, the global chromatin landscape and its molecular dynamics in this period remain largely unexplored in humans. Here we investigate chromatin states in human preimplantation development using an improved assay for transposase-accessible chromatin with high-throughput sequencing (ATAC-seq) 2 . We find widespread accessible chromatin regions in early human embryos that overlap extensively with putative cis-regulatory sequences and transposable elements. Integrative analyses show both conservation and divergence in regulatory circuitry between human and mouse early development, and between human pluripotency in vivo and human embryonic stem cells. In addition, we find widespread open chromatin regions before zygotic genome activation (ZGA). The accessible chromatin loci are readily found at CpG-rich promoters. Unexpectedly, many others reside in distal regions that overlap with DNA hypomethylated domains in human oocytes and are enriched for transcription factor-binding sites. A large portion of these regions then become inaccessible after ZGA in a transcription-dependent manner. Notably, such extensive chromatin reorganization during ZGA is conserved in mice and correlates with the reprogramming of the non-canonical histone mark H3K4me3, which is uniquely linked to genome silencing3-5. Taken together, these data not only reveal a conserved principle that underlies the chromatin transition during mammalian ZGA, but also help to advance our understanding of epigenetic reprogramming during human early development and in vitro fertilization.

Targeting macrophagic PIM-1 alleviates osteoarthritis by inhibiting NLRP3 inflammasome activation via suppressing mitochondrial ROS/Cl- efflux signaling pathway.

BACKGROUND: Osteoarthritis (OA), in which macrophage-driven synovitis is considered closely related to cartilage destruction and could occur at any stage, is an inflammatory arthritis. However, there are no effective targets to cure the progression of OA. The NOD-, LRR-,and pyrin domain-containing protein 3 (NLRP3) inflammasome in synovial macrophages participates in the pathological inflammatory process and treatment strategies targeting it are considered to be an effective approach for OA. PIM-1 kinase, as a downstream effector of many cytokine signaling pathways, plays a pro-inflammatory role in inflammatory disease. METHODS: In this study, we evaluated the expression of the PIM-1 and the infiltration of synovial macrophages in the human OA synovium. The effects and mechanism of PIM-1 were investigated in mice and human macrophages stimulated by lipopolysaccharide (LPS) and different agonists such as nigericin, ATP, Monosodium urate (MSU), and Aluminum salt (Alum). The protective effects on chondrocytes were assessed by a modified co-culture system induced by macrophage condition medium (CM). The therapeutic effect in vivo was confirmed by the medial meniscus (DMM)-induced OA in mice. RESULTS: The expression of PIM-1 was increased in the human OA synovium which was accompanied by the infiltration of synovial macrophages. In vitro experiments, suppression of PIM-1 by SMI-4a, a specific inhibitor, rapidly inhibited the NLRP3 inflammasome activation in mice and human macrophages and gasdermin-D (GSDME)-mediated pyroptosis. Furthermore, PIM-1 inhibition specifically blocked the apoptosis-associated speck-like protein containing a CARD (ASC) oligomerization in the assembly stage. Mechanistically, PIM-1 inhibition alleviated the mitochondrial reactive oxygen species (ROS)/chloride intracellular channel proteins (CLICs)-dependent Cl- efflux signaling pathway, which eventually resulted in the blockade of the ASC oligomerization and NLRP3 inflammasome activation. Furthermore, PIM-1 suppression showed chondroprotective effects in the modified co-culture system. Finally, SMI-4a significantly suppressed the expression of PIM-1 in the synovium and reduced the synovitis scores and the Osteoarthritis Research Society International (OARSI) score in the DMM-induced OA model. CONCLUSIONS: Therefore, PIM-1 represented a new class of promising targets as a treatment of OA to target these mechanisms in macrophages and widened the road to therapeutic strategies for OA.

Ultrafast dynamics of spin relaxation in monolayer WSe2 and the WSe2/graphene heterojunction.

Excitonic devices based on two-dimensional (2D) transition metal dichalcogenides (TMDCs) can combine spintronics with valleytronics due to its special energy band structure. In this work, we studied the generation and relaxation processes of spin/valley polarized excitons dynamics in monolayer WSe2 and its van der Waals (vdW) heterojunction with graphene using a circularly polarized femtosecond pump-probe system. The spin/valley depolarization dynamics of the A exciton in monolayer WSe2 is found to exhibit a biexponential decay. The fast relaxation process is due to the ultrafast intervalley electron-hole spin-flip exchange coupling and electron-phonon scattering. And the slow relaxation process originates from the recombination and relaxation of the trion states. Graphene has an electron extraction effect on WSe2, which prevents the formation of trions. Therefore, the spin/valley depolarization process of the A exciton in the heterojunction exhibits only a fast relaxation process. In both monolayer WSe2 and its heterojunction with graphene, B/A' excitons exhibit a negative spin/valley polarization which is mainly due to two-photon absorption and excited Bose scattering. Our work systematically studied the spin/valley depolarization dynamics of excitons and revealed possible mechanisms of their differences in isolated 2D WSe2 and vdW heterojunctions.

Association of glycated hemoglobin with non-alcoholic fatty liver disease patients and the severity of liver steatosis and fibrosis measured by transient elastography in adults without diabetes.

BACKGROUND: Type 2 diabetes mellitus (T2DM) is a well-known independent risk factor for non-alcoholic fatty liver disease (NAFLD). However, research exploring the association between blood glucose management and the risk of NAFLD status in subjects without diabetes was insufficient. This study aimed to explore the association of glycated hemoglobin (HbA1c) with NAFLD status and the severity of liver steatosis and fibrosis in non-diabetic people. METHODS: A cross-sectional analysis was conducted on 2998 non-diabetic American adults using data from the National Health and Nutrition Examination Survey (NHANES) 2017-2018 cycle. We used multivariable logistic regression models to evaluate the association between HbA1c and NAFLD status and the severity of liver steatosis and fibrosis. Interaction and stratified analyses were additionally performed. RESULTS: The multivariate regression analyses showed that HbA1c was associated independently with NAFLD status in all the models (model1: OR = 2.834, 95%CI: 2.321, 3.461; model 2: OR = 2.900, 95%CI: 2.312, 3.637 and model 3: OR = 1.664, 95%CI: 1.284, 2.156). We further performed the interaction and stratified analyses and discovered a significant interaction between HbA1c and BMI (Pinteraction < 0.05). Finally, a robust link was shown between HbA1c level and the severity of liver steatosis, which was mainly significant in the prediabetes group, while the correlation was not significant in HbA1c level and severity of liver fibrosis after controlling for all the potential confounders. CONCLUSIONS: We concluded that HbA1c level was positively correlated to the risk of developing NAFLD in a large non-diabetic American population. Moreover, HbA1c level was associated with the severity of liver steatosis in subjects with prediabetes, suggesting that routine screening for HbA1c among individuals with prediabetes is necessary.

MiR-601 Promotes Cell Proliferation of Human Glioblastoma Cells by Suppressing TINP1 Expression.

OBJECTIVE: It has been well documented that microRNAs (miRNAs) play essential roles in cancer initiation and development. In this study, we aimed to provide a better understanding of the mechanism of cell proliferation in glioblastoma (GBM). METHODS: Levels of miR-601 expression were detected in GBM tissues and cells. The effects of miR-601 dysregulation on GBM cell proliferation by mean transit time (brain tissue blood flow) (MTT) assays, colony formation, anchorage-independent growth assay, bromodeoxyuridine (BrdU) labeling and immunofluorescence assay. Bioinformatics analysis, luciferase reporter system and Western blot assays were used to predict and confirm the target gene miR-601. RESULTS: MiR-601 levels were identified as significantly up-regulated in GBM primary tumors and cell lines. Ectopic expression of miR-601 suppressed cell proliferation of GBM. Moreover, miR-601 showed its function by suppressing potential target TINP1 and TINP1 suppression reversed the effects of miR-601-in in U87MG GBM cells. CONCLUSION: Taken together, our results indicate that miR-601 promotes proliferation of GBM via inhibition of TINP1.

Three-Dimensional Crystalline Covalent Triazine Frameworks via a Polycondensation Approach.

The growth of crystalline covalent triazine frameworks (CTFs) is still considered as a great challenge due to the less reversible covalent bonds of triazine linkages. The research studies of crystalline CTFs to date have been limited to two-dimensional (2D) structures, and the three-dimensional (3D) crystalline CTFs have never been reported before. Herein we report the design and synthesis of two 3D crystalline CTFs, termed 3D CTF-TPM and 3D CTF-TPA through a reversible/irreversible polycondensation approach. The targeted 3D CTFs adopt ctn topology, and show moderate crystallinity, relatively large surface area (ca. 2000 m2 g-1 ), and high CO2 uptake capacity (23.61 wt.%). Moreover, these 3D CTFs exhibit ultrastability in the presence of boiling water, strong acid (1 M HCl) and strong base (1 M NaOH). This contribution represents the first report of 3D crystalline CTFs, which not only extends their structural diversity but also offers a synthetic strategy and structural basis for expanding practical applications of CTF materials.

Demystifying the Stern layer at a metal-electrolyte interface: Local dielectric constant, specific ion adsorption, and partial charge transfer.

Electric double layer (EDL) represents one of the most basic concepts in electrochemistry and is pertinent to diverse engineering applications ranging from electrocatalysis to energy storage. Whereas phenomenological and coarse-grained models have been long established to describe ionic distributions in the diffuse layer, a faithful prediction of the physicochemical properties of the electrode-electrolyte interface from a molecular perspective remains a daunting challenge. In this work, we investigate the charging behavior of an Ag (111) electrode in NaF aqueous solutions leveraging experimental results and theoretical calculations based on the classical density functional theory for ion distributions in the diffuse layer and on the joint density functional theory (JDFT) for the electronic structure. When the Ag electrode is applied with a negative voltage, the surface charge density can be reasonably described by assuming a neutral Stern layer with the dielectric constant dependent on the local electric field as predicted by the Kirkwood equation. However, the specific adsorption of F- ions must be considered when the electrode is positively charged and the fluoride adsorption can be attributed to both physical and chemical interactions. Qualitatively, F- binding and partial charge transfer are supported by JDFT calculations, which predict an increased binding energy as the voltage increases. Our findings shed insight on the molecular characteristics of the Stern layer and the charge behavior of adsorbed species not specified by conventional EDL models.

The role of UhpA in regulating the virulence gene expression in Edwardsiella piscicida.

Edwardsiella piscicida (E. piscicida) is an important fish pathogen. However, the mechanism of Glu6P transport regulatory protein UhpA how to affect the virulence gene expression in E. piscicida is still unclear. The results in this study showed that the metabolism-related gene expression of cysteine synthase (orf 1134) and sulphate transporter (ychM) in the uhpA mutant strain ΔuhpA was 0.76-fold and 0.68-fold lower than the ones in the wild strains (p < .05). The gene expression of ethA and ethB in the ΔuhpA strain was 0.80-fold and 0.72-fold lower than the ones in the wild strains (p < .05). However, the gene expression of fliC and flgN in the ΔuhpA was 1.51-fold and 1.21-fold higher than the ones in the wild strains (p < .05). The gene expression of T3SS (esrB and esrC) and T6SS (evpB and evpC) in the ΔuhpA was 1.27-fold, 1.13-fold, 1.28-fold and 1.23-fold higher than the ones in the wild strains (p < .05). This suggested that the uhpA gene could regulate the key virulence gene expression, and the uhpA gene was associated with the pathogenicity of E. piscicida in fish.

Risk of dietary intake of organochlorine pesticides among the childbearing-age women: A multiple follow-up study in North China.

Exposure to organochlorine pesticides (OCPs) can cause adverse health effects in the female population. We investigated the dietary OCP intake of childbearing-age women living in large agricultural areas of Northern China, as well as their associated health risks. Ten childbearing-age women were recruited during 2015-2016. Their weekly dietary intake diaries and food samples were collected over the course of five visits. The OCP residues of 322 food samples from seven categories (i.e., cereal, vegetable, fruit, fish, meat, egg, and milk) were analyzed by gas chromatography-mass spectrometry. The average concentrations of the total hexachlorocyclohexanes (ΣHCH), dichlorodiphenyltrichloroethanes and their metabolites (ΣDDX), endosulfans (ΣES), and dieldrin and endrin (ΣDrin) in all food categories were, overall, much lower than the maximum residue limits. Relative high mean residues of ΣDrin and ΣES were found in fruits (ΣDrin: 0.687 ng g-1 wet weight (w.w.), ΣES: 2.24 ng g-1 w.w.) and vegetables (ΣDrin: 0.690 ng g-1 w.w., ΣES: 2.11 ng g-1 w.w.). The estimated daily dietary intake (EDI) of these compounds was calculated, with mean levels of 10.6 (ΣES) > 4.37 (ΣDrin) > 1.51 (ΣHCH) > 0.850 (ΣDDX) ng kg-1 day-1. Women during the heating period (from January to March) tended to ingest more ΣHCH, ΣDDX, ΣDrin, and ΣES. Overall, women had no obvious non-carcinogenic and carcinogenic risks due to intake of OCPs, but 83.9% of them has potential carcinogenic risk, with estimated life carcinogenic risk (LCR) exceeding 10-6. Furthermore, women had a higher potential carcinogenic risk during the heating period (mean LCR: 1.33 × 10-5) than during the non-heating period (mean LCR: 8.50 × 10-6). ΣDrin was the dominant OCP responsible for health risks, followed by ΣHCH. We concluded that women in North China still have some dietary OCP intake, especially during the heating period.

Experimental Observation of the Gate-Controlled Reversal of the Anomalous Hall Effect in the Intrinsic Magnetic Topological Insulator MnBi2Te4 Device.

Magnetic topological insulator, a platform for realizing quantum anomalous Hall effect, axion state, and other novel quantum transport phenomena, has attracted a lot of interest. Recently, it is proposed that MnBi2Te4 is an intrinsic magnetic topological insulator, which may overcome the disadvantages in the magnetic doped topological insulator, such as disorder. Here we report on the gate-reserved anomalous Hall effect (AHE) in the MnBi2Te4 thin film. By tuning the Fermi level using the top/bottom gate, the AHE loop gradually decreases to zero and the sign is reversed. The positive AHE exhibits distinct coercive fields compared with the negative AHE. It reaches a maximum inside the gap of the Dirac cone, and its amplitude exhibits a linear scaling with the longitudinal conductance. The positive AHE is attributed to the competition of the intrinsic Berry curvature and the extrinsic skew scattering. Its gate-controlled switching contributes a scheme for the topological spin field-effect transistors.

Associations of Dietary Exposure to Organochlorine Pesticides from Plant-Origin Foods with Lipid Metabolism and Inflammation in Women: A Multiple Follow-up Study in North China.

This study explored effects of dietary OCP intake from plant-origin foods (cereals, fruits, and vegetables) consumption on lipid metabolism and inflammation of women using a multiple follow-up study. The results showed that dietary intake of p,p'-dichlorodiphenyltrichloroethane (DDT) [ß = - 10.11, 95% confidence interval (95%CI): - 17.32, - 2.905] and o,p'-dichlorodiphenyldichloroethylene (DDE) (ß = - 6.077, 95%CI: - 9.954, - 2.200) were overall negatively associated with serum high-density lipoprotein cholesterol (HDL), whereas other OCPs were not. Serum interleukin (IL)-8 was positively associated with intake of dieldrin (ß = 0.390, 95%CI: 0.105, 0.674), endosulfan-ß (ß = 0.361, 95%CI: 0.198, 0.523), total endosulfan (ß = 0.136, 95%CI: 0.037, 0.234), and total OCPs (ß = 0.084, 95%CI: 0.016, 0.153), and negatively correlated with intake of p,p'-DDE (ß = - 2.692, 95%CI: - 5.185, - 0.198). We concluded that dietary intake of some individual DDT-, DDE- dieldrin-, and endosulfan-class chemicals from plant-origin foods may interfere with lipid metabolism and inflammation responses.

Pioglitazone alleviates cisplatin nephrotoxicity by suppressing mitochondria-mediated apoptosis via SIRT1/p53 signalling.

Pioglitazone (PIO) attenuates cisplatin nephrotoxicity whereas the underlying mechanism remains unknown. Apoptosis is associated with mitochondrial dysfunction and SIRT1 activation can decrease cell apoptosis in cisplatin nephrotoxicity. Therefore, we explored whether the protective effect of PIO in cisplatin nephrotoxicity is achieved by suppressing mitochondria-mediated apoptosis through SIRT1/p53 signalling regulation. Cell viability, apoptosis, survival rate, renal pathology and function were examined. Moreover, we also analysed the expression of SIRT1, Acetyl-p53, mitochondrial membrane potential (MMP), reactive oxygen species (ROS), mitochondrial permeability transition pore (mPTP) opening, adenosine triphosphate (ATP) and apoptosis-related protein in vivo and in vitro. Pioglitazone treatment significantly increased cell viability, promoted SIRT1-p53 interaction, upregulated Bcl-2 expression, activated SIRT1 and elevated mitochondrial ATP synthesis after cisplatin treatment. However, PIO decreased the generation of ROS, opening of mPTP, dissipation of MMP and translocation of cytochrome c after cisplatin treatment. Pioglitazone also reduced the activation of caspase-3 and caspase-9, lowered the ratio of Bax/Bcl-2, attenuated kidney pathological damage and dysfunction, down-regulated the expression of Acetyl-p53, PUMA-α and Bax and abated cell apoptosis after cisplatin treatment. The SIRT1 inhibitor, EX527, clearly reversed the protective effects of PIO. These results implied PIO attenuated cisplatin nephrotoxicity by suppressing mitochondria-mediated apoptosis through regulating SIRT1/p53 signalling.

LuxS/AI-2 Quorum Sensing System in Edwardsiella piscicida Promotes Biofilm Formation and Pathogenicity.

LuxS/AI-2 is an important quorum sensing system which affects the growth, biofilm formation, virulence, and metabolism of bacteria. LuxS is encoded by the luxS gene, but how this gene is associated with a diverse array of physiological activities in Edwardsiella piscicida (E. piscicida) is not known. Here, we constructed an luxS gene mutant strain, the △luxS strain, to identify how LuxS/AI-2 affects pathogenicity. The results showed that LuxS was not found in the luxS gene mutant strain, and this gene deletion decreased E. piscicida growth compared to that of the wild-type strain. Meanwhile, the wild-type strain significantly increased penetration and motility in mucin compared to levels with the △luxS strain. The 50% lethal dose (LD50) of the E. piscicida △luxS strain for zebrafish was significantly higher than that of the wild-type strain, which suggested that the luxS gene deletion could attenuate the strain's virulence. The AI-2 activities of EIB202 were 56-fold higher than those in the △luxS strain, suggesting that the luxS gene promotes AI-2 production. Transcriptome results demonstrated that between cells infected with the △luxS strain and those infected with the wild-type strain 46 genes were significantly differentially regulated, which included 34 upregulated genes and 12 downregulated genes. Among these genes, the largest number were closely related to cell immunity and signaling systems. In addition, the biofilm formation ability of EIB202 was significantly higher than that of the △luxS strain. The supernatant of EIB202 increased the biofilm formation ability of the △luxS strain, which suggested that the luxS gene and its product LuxS enhanced biofilm formation in E. piscicida All results indicate that the LuxS/AI-2 quorum sensing system in E. piscicida promotes its pathogenicity through increasing a diverse array of physiological activities.

Interfacial Capillary-Force-Driven Self-Assembly of Monolayer Colloidal Crystals for Supersensitive Plasmonic Sensors.

Colloidal lithography technology based on monolayer colloidal crystals (MCCs) is considered as an outstanding candidate for fabricating large-area patterned functional nanostructures and devices. Although many efforts have been devoted to achieve various novel applicatons, the quality of MCCs, a key factor for the controllability and reproducibility of the patterned nanostructures, is often overlooked. In this work, an interfacial capillary-force-driven self-assembly strategy (ICFDS) is designed to realize a high-quality and highly-ordered hexagonal monolayer MCCs array by resorting the capillary effect of the interfacial water film at substrate surface as well as controlling the zeta potential of the polystyrene particles. Compared with the conventional self-assembly method, this approach can realize the reself-assembly process on the substrate surface with few colloidal aggregates, vacancy, and crystal boundary defects. Furthermore, various typical large-scale nanostructure arrays are achieved by combining reactive ion etching, metal-assisted chemical etching, and so forth. Specifically, benefiting from the as-fabricated high-quality 2D hexagonal colloidal crystals, the surface plasmon resonance (SPR) sensors achieve an excellent refractive index sensitivity value of 3497 nm RIU-1 , which is competent for detecting bovine serum albumin with an ultralow concentration of 10-8 m. This work opens a window to prepare high-quality MCCs for more potential applications.

The role of uhpA in Edwardsiella piscicida and the inflammatory cytokine response in tilapia.

Edwardsiella piscicida (E. piscicida) is an important zoonotic pathogen that infects fish by colonizing the intestines. The intestine provides nutrition including Glucose 6-phosphate (Glu6P) and a competitive environment for the microbiota. Although the transport system regulatory protein gene uhpA has been reported in E. piscicida genomes, whether the uhpA gene is involved in the pathogenicity of E. piscicida remains largely unknown. Therefore, the uhpA gene mutants strain E. piscicida ΔuhpA was constructed to elucidate the functions of Glu6P and the uhpA gene in E. piscicida. The results demonstrated that Glu6P significantly increased the gene expression of uhpC/uhpB/uhpA than without adding Glu6P in the culture. The gene expression of uhpC and uhpB was down regulated in the mutant strain than that of in the wild type strain. E. piscicida ΔuhpA exhibited an increase in virulence compared to that of E. piscicida EIB202 [LD50 value: (3.98 × 106 CFU/fish) and LD50 value: (1.45 × 107 CFU/fish) respectively]. Besides, although TNF-α did not show significant differences (p > 0.05) in the spleen of tilapia infected with ΔuhpA and EIB202 in the whole observed period, the gene expression of IL-1ß and TGF-ß in the spleen of tilapia infected with ΔuhpA showed significantly higher (p < 0.05) than that of in tilapia infected with EIB202. Meanwhile, the gene expression of IL-1ß and TGF-ß in spleen of tilapia infected with ΔuhpA showed significantly higher (p < 0.05) than that of in fish infected with EIB202 when zebrafish used as the control in the whole observed period. All these results suggested that Glu6P up-regulated the gene expression of uhpC/uhpB/uhpA; most important, the uhpA gene deletion in E. piscicida down-regulated the gene expression of uhpC and uhpB, enhanced its pathogenicity and its role in inducing the inflammatory cytokine responses in tilapia.

The UhpA mutant of Edwardsiella piscicida enhanced its motility and the colonization in the intestine of tilapia.

Edwardsiella piscicida (E. piscicida) is a significant bacterial pathogen of cultured fish, which infected fish meanly through the intestine. Glucose 6-phosphate (Glu6P) in the intestine is nutritious to the pathogen, Meanwhile, Glu6P was found using as a virulent regulating signal for bacteria. The UhpA, one of the Glu6P transport system regulatory proteins could down-regulate the uhpC/uhpB/uhpA system and decrease its pathogenicity. However, the motility and the colonization of E. piscicida affected by UhpA were still unclear. In this study, the motility and the colonization of E. piscicida were monitored. The result demonstrated that the motility of EIB202 was significantly stronger than that of in ΔuhpA according to fractions 4, 8 and 9. However, the motility of ΔuhpA was significantly stronger than that of EIB202 according to the total number at the whole experiment. Although, there was no difference in the number of bacteria in the posterior intestine of tilapia after infected with E. piscicida EIB202 and ΔuhpA. The number of bacteria in the anterior and the middle intestine of fish infected with ΔuhpA were significantly higher than that of in fish infected with EIB202 at the whole experiment (P < 0.05). Interestingly, both E. piscicida strains colonized in the anterior intestine than that of in the middle and posterior intestines of tilapia. Besides, the gene expression of IL-1ß and TNF-α in the head-kidney of fish infected with ΔuhpA showed significantly higher (p < 0.05) than fish infected with EIB202 during the whole experimental period. Most importantly, the survival rate of E. piscicida EIB202 and ΔuhpA were 57% and 37% respectively. All results indicate that the uhpA gene mutant in E. piscicida could enhance its motility and the colonization in the intestine of tilapia, this illustrates the mechanism of UhpA decreases the pathogenesis of E. piscicida in fish.

Cross-immunity in Nile tilapia vaccinated with Streptococcus agalactiae and Streptococcus iniae vaccines.

Streptococcus agalactiae and Streptococcus iniae are major bacterial pathogens of tilapia that can cause high mortality concomitant with large economic losses to aquaculture. Although development of vaccines using formalin-killed bacteria to control these diseases has been attempted, the mechanism of immunity against streptococcal infections and the cross-protective ability of these two bacteria remains unclear. To explore the immunological role of these vaccines, we compared the immune responses of tilapia after immunization with both vaccines and compared the relative percent survival (RPS) and cross-immunization protection of tilapia after separate infection with S. agalactiae and S. iniae. All results revealed that vaccinated fish had significantly higher (P < 0.05) levels of specific antibodies than control fish 14 days post secondary vaccination (PSV) and 7 days post challenge. In vaccinated fish, the mRNA expression of interleukin-8 (IL-8), interleukin-12 (IL-12), caspase-3 (C-3), tumour necrosis factor (TNF), and interferon (IFN) was significantly up regulated (P < 0.05) in the head kidney after immunized; similar results were found for IL-8, TNF and IFN in the posterior kidney, meanwhile the expression levels of C-3 and IFN were significantly increased (P < 0.05) in the spleen of vaccinated fish. Additionally, the levels of acid phosphatase (ACP), alkaline phosphatase (AKP), superoxide dismutase (SOD), and lysozyme (LZM) in vaccinated fish were improved at different degree when compared to the control fish. These results showed that vaccination with formalin-killed cells (FKCs) of either S. agalactiae or S. iniae conferred protection against infection by the corresponding pathogen in Nile tilapia, resulting in RPS values of 92.3% and 91.7%, respectively. Furthermore, cross-protection was observed, as the S. agalactiae FKC vaccine protected fish from S. iniae infection, and vice versa. These results suggested that the S. agalactiae and S. iniae FKC vaccines can induce immune responses and generate excellent protective effects in Nile tilapia.