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关键词： Mycobacterium tuberculosis   vitamin D   TNF   apoptosis   inflammation  

摘要： Vitamin D (VD) has been extensively associated with the resistance against tuberculosis (TB);however, the mechanism underlying the reduction in TB susceptibility by VD remains uncertain. In our prior investigation, we discovered the relationship between VD and mycobacterium tuberculosis ***-induced aberrant osteoclastogenesis. Here we report that VD diminishes apoptosis in ***-infected THP-1 cells through tumor necrosis factor (TNF) signaling pathway. This novel perspective contributes to the elucidation of the intricate relationship between VD and tuberculosis. In this study, THP-1 cells were infected with the Mycobacterium tuberculosis H37Rv strain (***) for 4h at a MOI of 1 and then treated with 1,25-dihydroxy vitamin D (1,25(OH)2D3) (10-6, 10-8, 10-10M) for 1d respectively. RNA sequencing (RNA-seq) was performed, and differential expression analysis was conducted by the R package edgeR. Immunofluorescence (IF) and immunohistochemistry (IHC) techniques were employed for VDR, TNFR1 and TUNEL in TB patients and serum levels of TNF-alpha and IL6 were measured simultaneously. Furthermore, the utilization of western blot and qRT-PCR techniques was employed to investigate the impact of VD on pivotal molecules involved in the TNF signaling pathway. In addition, Bacillus Calmette-Gu & eacute;rin (BCG, ATCC 35734, derived from ***) and VD were administrated by tail vein and articular cavity injection in vivo. Our findings revealed a robust responsiveness of the TNF signaling pathway to ***-induced inflammation, resulting in elevated expression of TNF-alpha, IL-6, and severe apoptosis. VD exhibited significant inhibitory effect on ***-induced inflammation and apoptosis both in vitro and in vivo. This study offers novel insights for vitamin D in the study of tuberculous bone destruction.

关键词： Gastric cancer stem cells   Clofoctol   SerpinE1/beta-catenin   RanBP2   Necroptosis  

摘要： Gastric cancer stem cells (GCSCs) play a crucial role in the initiation, progression, recurrence and therapeutic resistance, contributing to a poor prognosis. Consequently, GCSCs are deemed to be a potential therapeutic target for gastric cancer (GC). Although beta-catenin is a well-recognized therapeutic target for GC and several inhibitors have demonstrated potent anti-tumor effects, there is a dearth of therapeutic agents targeting beta-catenin for clinical therapy. In this study, we carried out high-throughput screening of clinically approved drugs to identify effective inhibitors of beta-catenin. The results revealed that the antibiotic drug, clofoctol (CFT) effectively reduced the beta-catenin level, attenuated stemness traits both in vitro and in vivo, and induced necroptosis of GCSCs. Further analyzing of downstream genes and targeted proteins, we found that CFT inhibited GCSCs viability by binding to the SUMO E3 ligase RanBP2, thereby suppressing the SerpinE1/beta-catenin axis and activating TNF-mediated necroptosis. These results indicate that CFT may exert potent therapeutic effects against GC by targeting beta-catenin and inhibiting the viability of GCSCs.

关键词： generalized pustular psoriasis   IL-17   IL-6   ixekizumab   proteomics  

关键词： 枝睾阔盘吸虫   脂肪酸结合蛋白   原核表达   多克隆抗体   ELISA  

摘要： 对枝睾阔盘吸虫（Eurytremacladorchis）的脂肪酸结合蛋白（FABP）进行原核表达与鉴定，并制备鼠抗EcFABP多克隆抗体。方法 通过PCR技术扩增了含开放阅读框（ORF）的EcFABP序列，构建重组表达质粒pET-28a-EcFABP。IPTG诱导EcFABP重组蛋白表达并进行最佳诱导时间优化。将纯化后的重组蛋白与弗氏佐剂等比例乳化后免疫BALB/c小鼠，制备多克隆抗体，经ELISA检测其抗体效价。同时采用Western blot检测多克隆抗体特异性，qPCR技术检测第42 d小鼠脾细胞因子（IL-4、IL-10、IL-12、IFN-γ）的转录水平。扩增出枝睾阔盘吸虫FABP基因ORF大小为402 bp，编码133个氨基酸，理论分子量约为15 kDa。SDS-PAGE和Western blot结果表明，本试验成功表达并纯化出EcFABP包涵体重组蛋白。间接ELISA法检测鼠源多克隆抗体，效价为1 256 000。Western blot结果表明，鼠抗EcFABP抗体可以与重组蛋白发生特异性反应。重组蛋白可诱导小鼠体内4种脾细胞因子表达上调，产生Th1和Th2两种类型的免疫反应。本试验成功制备了具有免疫原性的EcFABP蛋白及其鼠源多克隆抗体，为进一步研究枝睾阔盘吸虫FABP蛋白的功能及相关的诊断检测方法奠定了基础。

摘要： Immune evasion is a hallmark of gliomas, yet the genetic mechanisms by which tumors escape immune surveillance remain incompletely understood. In this study, we systematically examined the presence of somatic mutations in human leukocyte antigen (HLA) genes and genes encoding proteins involved in antigen-presentation across isocitrate dehydrogenase wild-type (IDHwt) and mutant (IDHmut) gliomas using targeted next-generation sequencing (NGS). To address the challenges associated with detecting somatic mutations in these highly polymorphic and complex regions of the genome, we applied a combination of short-read and long-read sequencing techniques, extended the genetic region of interest (exons and introns), and applied a tailored bioinformatics analysis pipeline, which enabled an accurate evaluation of comprehensive sequencing data. Our analysis identified mutations in HLA class II and non-classical HLA genes as well as genes associated with antigen presentation, such as TAP1/2 and B2M. Three-dimensional modeling of individual mutations simulated the potential impact of somatic mutations in TAP1 and B2M on the encoded protein configuration. The presence of somatic mutations supports the role of antigen-presenting genes in the pathophysiology and potential immune escape of gliomas. Our data demonstrated an increased frequency of such mutations in recurrent glioblastoma (GBM), potentially resulting from a positive selection or mutagenic enrichment of tumor cells during tumor progression. Taken together, this research generates new insights and hypotheses for the functional analysis and optimization of immunotherapy strategies for gliomas, which may guide personalized treatment paradigms.

关键词： Miocene  

摘要： Records from the Miocene il-Blata section in Malta offer insights into the depositional environments of the Central Mediterranean following an Early Miocene restriction of the Mesopotamian Seaway (c. 20 Ma). Inorganic and organic stable carbon isotope values suggest relatively steady depositional environments, whereas authigenic iron dolomite abundances exhibiting substantial delta O-18 and delta C-13 variations at a sequence scale indicate dynamic sedimentation conditions leading to differential diagenesis. Petrographically, the dolomitic levels exhibit matrix-selective dolomitization, occasional silicification and phosphatization, and textural indicators pointing to subsurface microbial influences. These features collectively point to complex shallow burial diagenesis. In addition, the presence of framboidal pyrite and gypsum infilling foraminiferal chambers, along with the absence of large planktonic foraminifera, suggests the development of palaeoenvironmental stress imposed by a density-stratified water column affecting the pore waters. Towards the top of the studied succession, a shift from organic to siliceous deposits reflects water column perturbations possibly linked to changes in oceanic circulation associated with a temporary re-opening of the Mesopotamian Seaway. This study not only underscores the hydrochemical controls exerted by North African terrigenous fluxes over the Mediterranean, but also highlights the intricate interplay between shifting depositional environments and shallow burial diagenetic processes in shaping the geochemical and textural fabrics of authigenic mineral assemblages.

关键词： Chagas disease   HLA-DRB1   cardiomyopathy   cross-reactivity   T-cells  

摘要： Introduction Human infection with the protozoan Trypanosoma cruzi causes Chagas disease, which may lead to a deadly dilated cardiomyopathy resulting from T-cell-mediated inflammation. We found that specific HLA-DRB1 alleles (*0103, *0402, *1301, and *1302) that display the DERAA motif are linked to this severe clinical manifestation of Chagas *** We employed computational analysis, in vitro functional assays, and single-cell RNA sequencing to determine the response of CD4+ T cells from indeterminate (IND) and cardiac (CCC) Chagas patients to peptides selected on DERAA-bearing *** We observed that these alleles display binding affinity towards host-derived peptides with sequence similarity to parasite-derived proteins. These peptides can activate and induce proliferation of CD4+ T-cells from CCC, but not IND. Importantly, the magnitude of this response correlated with the severity of ventricular dysfunction and increased production of soluble factors associated with myocardial fibrosis. Analysis of differentially expressed genes (DEGs) in activated CD4+ T-cells from individuals with the DERAA-DRB1 alleles demonstrated a high expression of cytotoxic, chemotactic and proapoptotic genes, linking these cells with pathogenic functions. Finally, we observed the upregulation of genes that code for the host proteins that contain the potentially pathogenic peptides in the cardiac tissue of CCC, suggesting their involvement in *** Our findings highlight the ability of CD4+ T-cells from CCC patients to recognize and react to foreign and self-peptides, thereby emphasizing the importance of HLA-DRB1 alleles in the presentation of potentially pathogenic antigens and in the amplification of CCC pathology.

摘要： Keloids are common pathological scars resulting from previous trauma or inflammation. Interferon-gamma (IFN-gamma) has shown significant therapeutic effects when used alone or in combination with other agents. While IFN-gamma has been found to regulate ferroptosis in tumor cells, its ability to regulate ferroptosis in keloid fibroblasts (KFs) is unclear. Here, we have demonstrated a direct causal relationship between IFN-gamma levels and ferroptosis in KFs. To explore the intrinsic mechanism, we performed genome-wide RNA and proteomics sequencing and found that serpine2 was the most significantly downregulated gene in KFs after exogenous overexpression of IFN-gamma. Serpine2, which belongs to a family of serine protease inhibitors, has been shown to play an important role in fibrotic diseases. Therefore, we hypothesized that serpine2 is a downstream gene in the regulation of ferroptosis in KFs by IFN-gamma. Our results showed that serpine2 overexpression promotes collagen synthesis, which in turn promotes the proliferation, migration, and invasive functions of KFs. We further demonstrated that serpine2 overexpression promoted system Xc- transporter expression, cystine uptake, and glutathione synthesis, enhanced GPX4 activity;and inhibited reactive oxygen species generation. This resulted in a reduction in intracellular lipid peroxidation and the levels of its metabolite malondialdehyde, as well as inhibited ferroptosis in KFs. IFN-gamma reversed these effects of serpine2 overexpression. These results were largely confirmed in in vivo keloid models too. These findings imply that IFN-gamma not only directly induces ferroptosis in KFs but also enhances their sensitivity to ferroptosis by inhibiting the synthesis of SLC7A11 and SLC3A2 through downregulation of serpine2. In summary, we suggest that the serpine2-system Xc- axis is a promising therapeutic target for the treatment of keloids.

关键词： Interleukin-4-induced-1   AlphaFold2   molecular docking   molecular dynamics simulation   MM/GBSA   ADMET analysis  

摘要： Interleukin-4-induced gene 1 (IL4I1) is an L-phenylalanine oxidase. As the primary enzyme responsible for degrading tryptophan, IL4I1 generates indole metabolites and kynurenic acid, which act as crucial endogenous ligands to activate the aryl hydrocarbon receptor (AHR). This activation enhances tumor survivability while suppressing the body's anti-tumor immune response. Consequently, IL4I1 is now recognized as a promising new target for drug development in the realm of cancer immunomodulation. In this study, we employed a strategy combining AlphaFold2 with molecular dynamics (MD) simulations to model receptor conformations our docking model achieved a regression fit with an R2 coefficient of 0.34, providing a robust framework for structure-based virtual screening aimed at identifying potential IL4I1 inhibitors. We then applied this structure-based virtual screening method to a compound library. After further MD simulation and following Molecular Mechanics/Generalized Born Surface Area (MM/GBSA) calculation of binding free energy and ADMET analysis, five candidate IL4I1 inhibitors were obtained. This study provides an effective in silico approach for the identification of IL4I1 inhibitors and offers a valuable reference for the virtual screening of inhibitors targeting other proteins without known structures.

关键词： BaEV lentivirus   CAR-NK   CD80   CD80-41BBL-mbIL21 K562 cells   umbilical cord blood-derived NK cells  

摘要： Natural Killer (NK) cells are an important population of the immune system, and NK cell-based therapy has shown great potential in the treatment of cancers. However, to apply NK cells clinically, producing a large number of cells with high cytotoxicity remains a challenge. Current strategies focus on employing different irradiated feeder cells to stimulate NK expansion, maturation, and cytotoxicity. While co-stimulatory signals play critical roles in promoting NK cell proliferation and activating their functions, the exploitation of these signals for expanding NK cells has not been fully explored. To identify the optimal engineered feeder cells for expanding umbilical cord blood-derived NK cells, we generated different feeder cells expressing the co-stimulatory molecules CD80, 4-1BBL, or membrane-bound IL-21 (mbIL21). We then evaluated the transduction efficacy of a chimeric antigen receptor (CAR) construct into expanded NK cells using various lentiviral vectors. Our results showed that CD80, in combination with 4-1BBL and mbIL21, induced the highest expansion of NK cells from cord blood. The expanded NK cells displayed higher cytotoxicity toward target cells compared to T cells following CAR transduction using BaEV lentivirus.