IRE1α-XBP1 regulates PDK1-dependent induction of epithelial-mesenchymal transition in non-small cell lung cancer cells.

Mounting evidence indicates that activation of unfolded protein response (UPR) and metabolic reprogramming contribute to cancer cell migration and invasion, but the molecular mechanism of pro-EMT program through a coordinated action of UPR with metabolism has not been defined. In this study, we utilized ER stress-inducing reagent, thapsigargin (TG), to induced pharmacologic ER stress in lung cancer cells. Here. We report that the branch of UPR, IRE1α-XBP1 pathway plays a pivotal role in reprogramming lung cancer cell metabolism. At the molecular level, the expression of pyruvate dehydrogenase kinase-1 (PDK-1) is directly induced by XBP1 as a consequence of UPR activation, thus facilitating aerobic glycolysis and lactate production. We also demonstrated that PDK1 serves as a downstream element of UPR activation in induction of Snail and EMT program. In addition, PDK1-induced Snail was dependent on the lactate production derived from metabolic reprogramming. Our findings reveal a critical role of lactate in pro-invasion events and establishes a direct connection between ER-stress and metabolic reprogramming in facilitating cancer cell progression.

Impairment of CRH in the intestinal mucosal epithelial barrier of pregnant Bama miniature pig induced by restraint stress.

Female, especially for pregnant female, are vulnerable to psychological stress. The morphology and metabolism of the maternal intestine are both obviously changed during pregnancy, thus making intestinal health status more fragile under psychological stress. The aim of the present study was to investigate the role of CRH and CRHR1 in the pregnant maternal intestine under psychological stress, thus exploring the mechanism of psychological stress in the pregnant maternal intestine. Bama miniature pigs were divided into the control and restraint stress groups from the first day of pregnancy. After restraint stress treatment for 18 consecutive days (D18), the plasma, duodenum, jejunum, ileum and colon were collected for study. Pregnant Bama miniature pigs subjected to restraint stress had significantly elevated CRH, adrenocorticotropic hormone (ACTH) and cortisol (COR) levels in plasma. Consistent with the increase in CRH levels, we observed enhanced oxidative stress levels in the intestine, which resulted in intestinal mucosal injury, including impaired intestinal morphology, a reduced number of goblet cells and proliferating cell nuclear antigen-positive cells, decreased expression of MUC2 and tight junctions, and elevated expression of CRHR1 and caspase-3. Moreover, exogenous CRH could directly promote IPEC-J2 cell apoptosis and influence its cell cycle (S and G2 phase) through CRHR1, and antalarmin could alleviate this phenomenon. Therefore, our results illustrated that the intestinal dysfunction of pregnant Bama miniature pigs was caused by restraint stress, and these changes were associated with the enhanced expression of CRH and CRHR1 in the intestine.

[Effect of equal concentration of Helicobacter pylori suspension on gastric mucosa in mice by different gavage methods].

OBJECTIVE: To investigate the effects of equal concentration of Helicobacter pylori suspension on gastric mucosal infection in mice by different gavage methods. 
 Methods: Six-week-old male C57BL/6 mice were infected by a suspension of Brucella broth containing the same amount of NCTC11637 Helicobacter pylori suspension by A, B, C, and D methods. For method A, the mice were intragastrically administered with Helicobacter pylori suspension (0.2 mL per mouse), once two day for 5 times; for method B, the mice were intragastrically administered with Helicobacter pylori (0.2 mL per mouse) once a day for 5 times; for method C, the mice were perfused with 0.4 mL per mouse of Helicobacter pylori suspension on the first day, then once a day and 0.2 mL per mouse for 3 times; for method D, the mice were administrated with 0.4 mL per mouse Helicobacter pylori suspension on the first day, 0.2 mL per mouse every other day for 3 times. For method E, the mice received equal amounts of normal saline. The mice were killed at 2, 4, and 6 weeks after gavage. The gastric mucosa was detected by rapid urease test for Helicobacter pylori infection, and gastric mucosa was taken for HE staining to observe the degree of infection.
 Results: After 2 weeks of gavage, the infection rates of the mice in A, B, C, and D group were 33.3%, 50.0%, 66.7%, and 33.3%, respectively. The degree of inflammation infection was as following order: C group>B group>D group>A group>E group. The infection rates of mice after 4 weeks of gavage in the A, B, C, and D groups were 50.0%, 83.3%, 83.3%, and 66.7%, respectively. The degree of inflammation infection was as following order: C group>B group>D group>A group>E group. After 6 weeks of gavage, the infection rate in A, B, C, and D groups was 100%, while the degree of inflammation infection was as following order: C group>D group>B group>A group>E group.
 Conclusion: At the acute stage of Helicobacter pylori infection, different gavage methods show different infection rates in mice, and the degree of inflammation is different. At the chronic stage, different gavage methods display the same infection rate in mice with different degree. The gavage method that 0.4 mL Helicobacter pylori suspension on the first day, then once a day and 0.2 mL for 3 times is most conducive to Helicobacter pylori colonization in the gastric mucosa of mice. This method can induce the the most seriou inflammatory infection and is beneficial to the successful establishment of the Helicobacter pylori infection model.

Enterococci facilitate polymicrobial infections.

Enterococci are ubiquitous members of the gut microbiota in human beings and animals and are among the most important nosocomial organisms. Due to their opportunistic pathogenicity, enterococci are referred to as pathobionts and play decisive roles in a diverse array of polymicrobial infections. Enterococci can promote the colonization, pathogenesis, and persistence of various pathogens, compromise the efficacy of drugs, and pose a severe threat to public health. Most current treatments tend to focus on the sole pathogenic bacteria, with insufficient attention to the driving role of enterococci. In this review, we summarize the characteristics of enterococci in infections, the factors facilitating their outgrowth, as well as the sites and types of enterococci-associated polymicrobial infections. We present an overview of the underlying mechanisms of enterococci-mediated pathogenesis in polymicrobial infections. Furthermore, we discuss alternative strategies and potential intervention approaches to restrict such infections, shedding light on the discovery and development of new therapies against polymicrobial infections.

[Vitamin D3 alleviates the gastritis that associated with Helicobacter pylori infection in mice with hypercholesterolemia by enhancing the activity of vitamin D receptors in the liver tissue and blocking the signaling pathway of JAK/STAT3].

Objective To investigate whether vitamin D3 (VD3) can alleviate Helicobacter pylori (Hp) infection by reducing blood lipids and inhibiting the Janus kinase/signal transducer and activator of transcription 3 (JAK/STAT3) signaling pathway. Methods High-cholesterol mouse model and Hp infected mouse model were established. Each was treated with VD3 via oral administration for 8 weeks. Real-time quantitative PCR was used to detect the expression of vitamin D receptor (VDR), insulin-induced gene 2 (Insig-2), and gastrin mRNA. Western blot analysis was used to examine the expression of JAK, STAT3, and cyclooxygenase-2 (COX2) proteins in gastric tissues. Biochemical analyses were performed to measure serum cholesterol levels, and ELISA was utilized to evaluate serum gastrin, interleukin 6 (IL-6), and IL-8 levels, along with histopathological examination of liver and gastric tissues using HE staining. Results After oral administration of VD3, the levels of VDR and Insig-2 in mouse liver tissue significantly increased in the high cholesterol group and the high cholesterol combined with Hp infection group. And the expression of serum gastrin decreased. The expression of JAK, STAT3 in gastric tissues reduced, as did the expression of COX2. Serum cholesterol levels decreased, with no significant changes in IL-6 levels, but a reduction in IL-8 levels. Compared to the control group, the high cholesterol combined with Hp infection group showed reduced hepatic ballooning degeneration and alleviated gastric tissue inflammation. In addition, inflammation in gastric tissue was also reduced in the cholesterol group and the Hp infection group. Conclusion VD3 alleviates gastritis by enhancing the activity of VDR in liver tissues, blocking the JAK/STAT3 signaling pathway, and inhibiting the expression of inflammatory factors.

Coumarin Glycosides Reverse Enterococci-Facilitated Enteric Infections.

Commensal enterococci with pathogenic potential often facilitate the growth of diverse pathogens, thereby exacerbating infections. However, there are few effective therapeutic strategies to prevent and intervene in enterococci-mediated polymicrobial infections. Here, we find that enterococci at high density drive the expansion and pathogenicity of enteric Salmonella enterica serotype Typhimurium (S. Tm). Subsequently, we show that the driving role of enterococci in such infections is counteracted by dietary coumarin glycosides in vivo. Enterococci, which are tolerant of iron-deficient environments, produce ß-glucosidases to hydrolyze coumarin glycosides into bioactive aglycones, inhibiting S. Tm growth and ameliorating the severity of S. Tm-induced symptoms by inducing iron limitation. Overall, we demonstrate that coumarin glycosides as a common diet effectively reverse enterococci-facilitated enteric infections, providing an alternative intervention to combat polymicrobial infections.

Emerging Risks in Food: Probiotic Enterococci Pose a Threat to Public Health through the Food Chain.

Probiotics have been associated with clinical infections, toxicity, and antimicrobial resistance transfer, raising public concerns. Probiotic enterococci are emerging food risks as opportunistic pathogens, yet little attention has been paid to them. Herein, we collected 88 enterococcal isolates from probiotic products used for humans, companion animals, livestock, and aquaculture. Results showed that all 88 probiotic enterococcal isolates harbored diverse virulence genes, multiple antimicrobial resistance genes, and mobile genetic elements. Notably, 77 isolates were highly resistant to gentamicin. Representative enterococcal isolates exerted toxic activities in both in vitro and in vivo models. Collectively, our findings suggest that probiotic enterococci may be harmful to hosts and pose a potential threat to public health.

FOXO1 Is a Critical Switch Molecule for Autophagy and Apoptosis of Sow Endometrial Epithelial Cells Caused by Oxidative Stress.

Oxidative stress (OS) is involved in various reproductive diseases and can induce autophagy and apoptosis, which determine the different fates of cells. However, the sequence and the switch mechanism between autophagy and apoptosis are unclear. Here, we reported that chronic restraint stress (CRS) induced OS (decreased T-AOC, T-SOD, CAT and GSH-Px and increased MDA) and then disturbed the endocrine environment of sows during early pregnancy, including the hypothalamic-pituitary-ovarian (HPO) and the hypothalamic-pituitary-adrenal (HPA) axes. Meanwhile, after CRS, the KEAP1/NRF2 pathway was inhibited and attenuated the antioxidative ability to cause OS of the endometrium. The norepinephrine (NE) triggered ß 2-AR to activate the FOXO1/NF-κB pathway, which induced endometrial inflammation. CRS induced the caspase-dependent apoptosis pathway and caused MAP1LC3-II accumulation, SQSTM1/p62 degradation, and autophagosome formation to initiate autophagy. Furthermore, in vitro, a cellular OS model was established by adding hydrogen peroxide into cells. Low OS maintained the viability of endometrial epithelial cells by triggering autophagy, while high OS induced cell death by initiating caspase-dependent apoptosis. Autophagy preceded the occurrence of apoptosis, which depended on the subcellular localization of FOXO1. In the low OS group, FOXO1 was exported from the nucleus to be modified into Ac-FOXO1 and bound to ATG7 in the cytoplasm, which promoted autophagy to protect cells. In the high OS group, FOXO1 located in the nucleus to promote transcription of proapoptotic proteins and then induce apoptosis. Here, FOXO1, as a redox sensor switch, regulated the transformation of cell autophagy and apoptosis. In summary, the posttranslational modification of FOXO1 may become the target of OS treatment.

Zeolitic-imidazolate frameworks derived Pt-free counter electrodes for high-performance quantum dot-sensitized solar cells.

Zeolitic-imidazole frameworks (ZIFs), as novel porous materials, are attracting much attention in several fields due to their special advantages such as large specific surface area, versatile porosity and well-connected networks. Here, we develop a porous ZIF-derived catalytic thin film, which was coated on the conducting glass as a counter electrode (CE) to substitute costly platinum for quantum dot-sensitized solar cells (QDSSCs). A ZIF layer is first prepared by coating ZIF-67 powders on the conducting glass, followed by the careful calcination treatments in sulfur vapour (sulfuration) or nitrogen gas (carbonization). The structure and morphologies of the derived porous film are characterized by the measurements of XRD, SEM and BET, and the electrochemical properties in the polysulfide solution are evaluated by the measurements of Tafel curves and electrochemical impedance spectroscopies. The derived porous film is used as a CE to fabricate QDSSC with CdSe quantum dot-sensitized TiO2 nanocrystalline thin film and the polysulfide solution. Compared with the photovoltaic performance of CdSe QDSSCs based on the CE prepared by the different sulfuration conditions, QDSSC based on the CE derived by the sulfuration for 30 min shows an excellent light-to-electric conversion efficiency of 3.77%, it is even higher than that of QDSSC based on Pt CE (2.98%). This work will open a new avenue to design a facile, low-cost and renewable CE for QDSSC.

Identification of plasma-responsive outer membrane proteins and their vaccine potential in Edwardsiella tarda using proteomic approach.

We have used differential sub-proteomic methodologies to detect Edwardsiella tarda outer membrane (OM) protein expression regulation during interaction with fish and human plasma, which is the critical step of the bacterial invasion internal organs via blood circulation. Seven and nine OM proteins were differentially expressed in response to fish and human plasma stress, respectively. Six proteins, TolB2, ETAE_2935, ETAE_0245, EvpA, ETAE_2675 and OmpA, were the shared proteins with the similar changes between the two plasma treatments. Except for EvpA, which was a known protein involved in bacterial pathogenesis and stress sensing, the others were first reported here to be related to bacterial invasion and infection. Out of them, four, upregulated ETAE_0245 and OmpA and downregulated ETAE_2675 and ETAE_2935, were selected for investigation of immune protection. The upregulated OmpA and ETAE_0245 were able to induce bactericidal antibodies in mice. These findings demonstrate that differential proteomic methodologies following protein expression regulation to interaction between host and pathogen with bacterial challenge post immunization of these altered proteins is a valid approach for identifying new vaccine candidates and nicely complements other high throughput mining strategies used for vaccine discovery.

Identification and network of outer membrane proteins regulating streptomysin resistance in Escherichia coli.

Bacterial Outer membrane (OM) proteins involved in antibiotic resistance have been reported. However, little is known about the OM proteins and their interaction network regulating streptomycin (SM) resistance. In the present study, a subproteomic approach was utilized to characterize OM proteins of Escherichia coli with SM resistance. TolC, OmpT and LamB were found to be up-regulated, and FadL, OmpW and a location-unknown protein Dps were down-regulated in the SM-resistant E. coli strain. These changes at the level of protein expression were validated using Western blotting. The possible roles of the altered proteins involved in the SM resistance were investigated using genetic modified strains with the deletion of these altered genes. It is found that decreased and elevated minimum inhibitory concentrations and survival capabilities of the gene deleted strains and their resistant strains, Delta tolC, Delta ompT, Delta dps, Delta tolC-R, Delta ompT-R, Delta dps-R and Delta fadL-R, were correlated with the changes of TolC, OmpT, Dps and FadL at the protein expression levels detected by 2-DE gels, respectively. The results may suggest that these proteins are the key OM proteins and play important roles in the regulation of SM resistance in E. coli. Furthermore, an interaction network of altered OM proteins involved in the SM resistance was proposed in this report. Of the six altered proteins, TolC may play a central role in the network. These findings may provide novel insights into mechanisms of SM resistance in E. coli.