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关键词： Selenium   T-2 toxin   Spleen injury   P38 MAPK   NF-kappa B   IL-6  

摘要： Both T-2 toxin and selenium (Se) cause immune impairment in the spleen, but the mechanism of their occurrence is not clear. In this experiment, the Se content of the spleen was tested by atomic fluorescence spectrometry after a 12-week intervention in Sprague-Dawley (SD) rats, divided into normal, normal + T-2 toxin (10 ng/g), normal + T-2 toxin (100 ng/g), low Se, low Se + T-2 toxin (10 ng/g), and low Se + T-2 toxin (100 ng/g) groups. The pathological changes and fibrosis of spleen tissue were observed using hematoxylin-eosin (HE) staining and Masson staining, respectively. Mitogen-activated protein kinase p38 (p38 MAPK), phosphorylated protein-38 (P-p38 MAPK), nuclear factor kappa-B (NF-kappa B), and interleukin-6 (IL-6) expression levels in spleen tissues were analyzed by Western blotting (WB) and immunohistochemistry (IHC) staining. This study found that both Se deficiency and T-2 toxin induced inflammatory injury and fibrotic changes in rat spleen, but low selenium and low selenium combined with T-2 toxin intervention showed more intense splenic injury. Se deficiency combined with T-2 toxin intervention aggravated spleen injury, and the mechanism of occurrence involved an increase in the inflammatory injury in the spleen by elevating the expression levels of p38 MAPK, NF-kappa B, and IL-6 in rat spleen.

关键词： DNA extraction   Microtube   PCR-SSOP   HLA typing  

摘要： Background The purity of the extracted DNA is critical for successful molecular testing. This study aimed to compare the effect of various DNA extraction methods, extraction processes, and sources of consumables, such as microtubes, on PCR results. Methods DNA extraction from whole blood was performed using four different approaches utilizing four types of microtubes: chloroform-based, sodium perchlorate-based, heat-assisted salting out, and solid-phase extraction. Extracted DNA was evaluated by nanodrop spectrophotometry and used in PCR-based methods for human leukocyte antigens (HLA) typing. Results The lowest and highest concentrations of extracted DNA were observed in the column-based and heat-assisted methods, respectively. The mean ratios of A260/A230, represents chemical contamination, were significantly lower in all extraction methods using all types of microtubes versus "A" microtube. We observed the highest mean ratio of A260/A230 (> 1.9) in the sodium perchlorate method by using the "A" microtube compared to other types of microtubes and extraction methods. Extracted DNA samples using microtube "A" showed acceptable results for HLA typing compared to other microtubes that represented uninterpretable results in the PCR using the sequence-specific oligonucleotide probe (PCR-SSOP) method. Conclusion Our findings indicate that the chemical contaminations derived mainly from microtubes can decrease the quality of DNA and consequently interfere with the amplification or hybridization reactions in the PCR-SSOP method for HLA typing. Although, technical issues and extraction processes may also influence the quality of DNA.

关键词： Chimeric antigen receptor   Env183-191 epitope   HBV-HCC   MHC   TCR-like antibody  

摘要： CAR-T cell therapy demonstrates significant efficacy in hematologic malignancies, with target selection critically determining therapeutic outcomes. However, the available tumor surface antigens are limited, especially in the treatment of solid tumors. A potential solution to overcome this limitation entails employing antibodies recognizing peptide-major histocompatibility complex (pMHC) structures, enabling CAR-T cell to detect intracellular tumor antigens through a T cell receptor (TCR)-like recognition mechanism. This study focuses on HBV-associated hepatocellular carcinoma (HBV-HCC), where HBV DNA integration into the host genome generates specific viral antigen epitopes presented by MHC class I molecules, representing attractive targets for CAR-T cell therapy. We engineered CAR-T cells with a TCR-like antibody (HBs183 CAR-T) specific for the immunodominant HBV envelope epitope Env183-191 presented by HLA-A *0201, and evaluated the antigen-specific cytotoxicity and safety profile of the CAR-T cells through in vitro functional assays and in vivo evaluation in heterogenous tumor models (subcutaneous and intraperitoneal xenografts). Our research provides a reference for CAR-T cell therapy targeting intracellular antigens, particularly specific antigens derived from viral infections, as targets for CAR-T treatment, and offers a preliminary concept validation for the CAR-T treatment of HBV-HCC tumors.

关键词： Renal ischemia/reperfusion   Remote ischemic pre-conditioning   Apelin   Inflammation   Oxidative stress  

摘要： Background Remote limb ischemic pre-conditioning (RIPreC) can invoke potent renal protection. The involvement of oxidative stress and inflammatory pathways in renal ischemia/reperfusion injury (I/RI) was also confirmed. This study was designed to investigate the RIPreC effects on IRI-induced kidney dysfunction in rats through NF kappa B/TNF-alpha/TGF-beta/apelin signaling *** Renal I/RI was induced by occluding the kidney arteries for 45 min, then reperfusion for 24 h. Four similar cycles of left femoral artery ischemia (2 min)/reperfusion (3 min) before the onset of kidney ischemia were performed to create RIPreC. Animals were randomly divided into three groups: sham, I/R, and RIPreC + I/R. Following the reperfusion phase, urine and blood samples were taken, and the kidney was removed for functional, molecular, and histological *** When compared to sham rats, renal IRI resulted in decreased creatinine clearance and increased sodium fractional excretion, lower antioxidant enzyme activities, higher malondialdehyde content and higher nuclear factor-kappa B (NF-kappa B), tumor necrosis factor-alpha (TNF-alpha), transforming growth factor-betta (TGF-beta), and Apelin expression levels, and histologically damaged kidney tissue. All of the alterations, as mentioned earlier, were alleviated using the RIPreC *** Thus, RIPreC can protect against renal dysfunction after renal I/RI via modulation of the TNF-alpha/NF-kappa B/TGF-beta Apelin signaling pathway and strengthening the antioxidant defense system.

关键词： COVID-19   HIV infections   Polymorphism   Single nucleotide   Interleukin   Cytokine storm  

摘要： Introduction The people living with HIV with abnormal immune responses have been identified as a population that is particularly susceptibility to contracting COVID-19. We explored the correlation between gene polymorphisms of IL-17A, IL-17F, and IL-6, and the susceptibility to COVID-19 in individuals with HIV infection. Methods In this cross-sectional study, 337 HIV-positive patients were included. Serological and molecular tests were done using ELISA and PCR-RFLP methods. Allelic frequency, haplotype analyses, linkage disequilibrium were calculated. A linear regression model was used to analyze the interleukin SNP genotypes in HIV patients with and without COVID-19. Results A total of 337 PLWH were recruited for this study, with 170 having COVID-19 and 167 not having it. The mean age and laboratory indicators showed no significant differences between the two groups (P > 0.05). The allele frequency analysis found no significant difference in the IL-17A rs2275913 polymorphism between case and control groups. However, the IL-17F rs763780 and IL-6 rs1800795 had significantly greater frequencies of specific alleles in the case group compared to the control group. The A-A haplotype of IL-17 in SNPs-rs 2,275,913 and rs763780 rising the risk of COVID-19 infection in PLWH by up to 2.398 times compared with the other haplotypes, and the A-G and G-A haplotypes have a protective role against the incidence of COVID-19 infection. Conclusion This study is the first to show a significant correlation between the prevalence of COVID-19 and variety polymorphism at IL-17 and IL-6, which suggests that genetic changes in interleukin genes may relate to COVID-19 distribution.

关键词： Interleukin-8   Topoisomerase I   Natural product   Taibai mountain  

摘要： This study aimed to screen IL-8/topo I inhibitors from natural products collected in Taibai mountain and explore the underlying mechanisms. A natural product library of Taibai mountain (NPTM) was first constructed containing 186 compounds. Then, 15 and 6 potential inhibitors were screened using pharmacophore modeling for IL-8 and topo I, respectively. Molecular docking indicated that glycycoumarin was IL-8/topo I inhibitor. A 200 ns molecular dynamics simulation reflected high binding stability and favorable hydrogen bond interaction within glycycoumarin-IL-8/topo I complexes. MM/GBSA calculation showed that the binding free energy was - 12.81 kcal/mol and - 31.20 kcal/mol, and Pro 32 and DT 10 contributed most to glycycoumarin and IL-8, topo I, respectively. SMD simulation demonstrated a stable binding under physiological conditions with energy demands to dissociate from IL-8/topo I. Glycycoumarin decreased IL-8 production in the inflammatory cells, and exhibited obvious topo I inhibition, as well as strong cytotoxicity to three cancer cells. A PPI network revealed that glycycoumarin might work through proteoglycans in cancer (hsa05205) and IL-17 signaling pathway (hsa04657). These results improved current understanding of natural IL-8/topo I inhibitors from Taibai mountain. The combination of in silico and bioassay could provide a new strategy for exploring natural lead compounds against inflammation and cancer.

关键词： Pyroptosis   Glucocorticoid resistance   IL-8   Epithelial cells  

摘要： ObjectiveThis study seeks to elucidate the role and molecular mechanisms of IL-8 in nasal epithelial cell pyroptosis and its impact on glucocorticoid (GC) *** assessed the expression of pyroptosis-related biomarkers and IL-8 in tissues and human nasal epithelial cells (hNECs) from both control and nasal polyp patients using western blot. Their localization was determined through immunohistochemistry and immunofluorescence. Cell death and cytotoxicity assay, electron microscopy, ELISA, and immunofluorescence were utilized to investigate IL-8-induced pyroptosis and GC resistance in hNECs, alongside the examination of the involved signaling pathways via western blot and immunofluorescence. In a murine model, hematoxylin-eosin staining and immunohistochemistry clarified relationship between pyroptosis and GC ***-8 and pyroptotic biomarker expression were significantly higher in nasal polyp-derived tissues and hNECs compared to controls. IL-8 showed a positive correlation and co-localized with the pyroptotic biomarkers. IL-8 triggered pyroptosis in hNECs by activating the ERK signaling pathway, leading to increased IL-1 beta and IL-18 secretion. Moreover, IL-8-induced pyroptosis was found to contribute to GC resistance by affecting phosphorylation of GC receptor Ser211. Inhibition of pyroptotic proteins mitigated IL-8-induced GC resistance both in vitro and in *** IL-8 facilitates pyroptosis via the ERK signaling pathway and plays a significant role in GC resistance in nasal polyps.

关键词： Conservation genomics   Immune genes   Bellinger River turtle   Major histocompatibility complex   Whole-genome re-sequencing  

摘要： Small, isolated populations are often vulnerable to increased inbreeding and genetic drift, both of which elevate the risk of extinction. The Bellinger River turtle (Myuchelys georgesi) is a critically endangered species endemic to a single river catchment in New South Wales, Australia. The only extant wild population, along with the breeding program, face significant threats from viral outbreaks, most notably a nidovirus outbreak in 2015 that led to a 90% population decline. To enhance our understanding of genomic characteristics in the species, including genome-wide and functional gene diversity, we re-sequenced, assembled, and analysed 31 re-sequenced genomes for pure M. georgesi (N = 31). We manually annotated the major histocompatibility complex (MHC), identifying five MHC class I and ten MHC class II genes and investigated genetic diversity across both classes in M. georgesi. Our results showed that genome-wide diversity is critically low in pure M. georgesi, contexualised through comparison with opportunistically sampled backcross animals-offspring of F1 hybrids (M. georgesi x Emydura macquarii) backcrossed to pure M. georgesi (N = 4). However, the variation observed within the core MHC region of pure M. georgesi, extending across scaffold 10, exceeded that of all other macrochromosomes. Additionally, no significant short-term changes in either genome-wide or immunogenetic diversity were detected following the 2015 nidovirus outbreak (before;N = 19, after;N = 12). Demographic history reconstructions indicated a sustained, long-term decline in effective population size since the last interglacial period, accompanied by more recent steep declines. These patterns suggested that prolonged isolation and reduced population size have significantly influenced the dynamics of genome-wide diversity. It is likely that contemporary stressors, including the recent nidovirus outbreak, are acting on an already genetically depleted population. This study offers new insights

关键词： Sepsis   Macrophage   ILC2   IL-9   Lung injury  

摘要： BackgroundGroup 2 innate lymphoid cells (ILC2) are the main group of tissue-resident ILCs in the lungs, which protect airway barrier integrity following infection. Macrophages are integral to the regulation of immune homeostasis in sepsis. However, the relationship between ILC2 and macrophages in the context of sepsis induced acute lung injury remains *** sepsis was conducted by cecal ligation and puncture (CLP) model in Wild Type (WT) mice and ILC2 depleted mice. Septic mice were injected intratracheally IL-9, and the frequency and markers expression of ILC2 and macrophage were measured by Flow cytometry and CyTOF. The lung injury was conducted with pathological analysis. In vitro studies, MH-S cells were exposed to LPS with/without interleukin-9 (IL-9), and mTOR level and MH-S cells death were measured with western bloting or Flow *** induced the accumulation of ILCs and pulmonary macrophages in lungs. Furtherly, we revealed that ILC2 and CD45+F4/80+CD11c+ macrophages expanded during sepsis induced acute lung injury. Meanwhile, ILC2 depletion significantly enhanced macrophages expansion. In vivo and in vitro studies determined that pulmonary macrophage death followed by sepsis were protedced by IL-9, which was main secreted by ILC2 in lung. Furthermore, IL-9 significantly declined the expression of mTOR, and the presence of ILC2 or IL-9 reduced the expression of M1 markers (CD86 or MHC II).ConclusionsIL-9 secreted by ILC2 has a protective role in sepsis induced lung injury by reducing macrophage apoptosis and M1 polarization via mTOR.

关键词： Hepatitis C   HCV   Cellular immune response   CD8(+ )T response   HLA-A*02:01   Peptide microarray  

摘要： Several barriers for the development of an HCV vaccine still exist, including the genetic diversity of the virus, and the shortage of assessable models for in vitro and in vivo assays. Therefore, in this study, HCV epitope mapping was performed for 59 polyprotein sequences from 7 HCV genotypes. Around 2,880 peptides were considered epitopes for CD8+ T cells. The peptide induction of cytokines from Th1 and/or Th2 axes of the cellular immune response was assessed, indicating a tendency for Th2 axis. In vitro evaluation was performed using peptide microarray and a recombinant HLA-A*02:01 molecule. A total of 615 peptides of high reactivity to HLA-A*02:01 were identified, with predominance of leucine and tryptophan residues, highlighting their importance for TCR-epitope binding and CD8+ T activation. Finally, HCV-derived peptide patterns restricted to HLA-A2*02:01 observed in this study provide important information for the development of a multi-epitope-based pan-genotypic vaccine against the virus.