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关键词： 补阳还五汤   坏死性凋亡   心肌梗死   心室重构  

摘要： 目的：探索补阳还五汤对TNF/RIPK1通路的影响，阐明补阳还五汤抑制心肌梗死后心室病理性重构的作用机制。方法：将70只Wistar大鼠随机分为对照组，模型组，补阳还五汤低、中、高剂量组，西药组以及TNF-α 抑制剂组，每组10只。补阳还五汤低、中、高剂量组分别给予补阳还五汤3.22 g·kg-1·d-1、 6.44 g·kg-1·d-1、12.87 g·kg-1·d-1，西药组给予培哚普利叔丁胺片0.36 mg·kg-1·d-1，TNF-α 抑制剂组给予TNF-α 抑制剂依那西普5 mg·kg-1·w-1皮下注射，对照组与模型组均给予生理盐水灌胃，连续给药4周。采用多普勒超声检测各组大鼠心脏的左室射血分数（ejection fraction of left ventricular， LVEF）、左室短轴缩短率（left ventricular fractional shortening， FS）、左室舒张末期内径（left ventricular end diastolic diameter， LVEDD）和左室收缩末期内径（left ventricular end systolic diameter， LVESD）。病理切片进行HE染色和Masson染色观察心肌组织的病理形态及纤维化程度；Tunel染色观察心肌细胞凋亡情况。RT-qPCR检测心肌组织中TNF-α、RIPK1、RIPK3、MLKL的mRNA表达水平。Western blot检测心肌组织中TNF-α、RIPK1、P-RIPK1、RIPK3、MLKL的蛋白表达水平。结果：与对照组相比，模型组大鼠的正常心脏结构被破坏、室壁变薄、心肌细胞排列紊乱、局部出现纤维化重构，LVEF、FS明显降低，LVEDD、LVESD明显升高，TNF-α、RIPK1、RIPK3、MLKL的mRNA表达和TNF-α、P-RIPK1、RIPK3、MLKL的蛋白表达明显升高（P<0.05）；与模型组比较，补阳还五汤低、中、高剂量组、西药组以及TNF-α 抑制剂组大鼠的心脏结构破坏程度和心肌纤维化程度均有所改善，LVEF、FS明显增高，LVEDD、LVESD明显降低，TNF-α、RIPK1、RIPK3、MLKL的mRNA表达水平和TNF-α、P-RIPK1、RIPK3、MLKL的蛋白表达水平降低（P<0.05），且补阳还五汤高剂量组疗效最好。结论：补阳还五汤可能通过抑制TNF/RIPK1通路减少心肌细胞坏死性凋亡从而改善心肌梗死后心室病理性重构。

关键词： salivary glands   Sjogren's disease   tissue dissociation   IFN pathway cytometry   flow cytometry  

摘要： Aim/Introduction We aimed to achieve direct quantitative measurement of activated and therapeutically actionable pathways (e.g., Type-I interferon) in target organs of autoimmune disease using flow cytometry of human salivary glands. Sj & ouml;gren's Disease (SjD) is a systemic autoimmune disorder characterized by lymphocytic inflammation and dysfunction of the lacrimal and salivary glands. Minor salivary glands are routinely biopsied and are used for the histopathological diagnosis of SjD. In this study, we optimized the dissociation, permeabilization, antibody panel, and analytical parameters to characterize both the immune and epithelial cells in the glands, and the activation status of a specified pathway by measuring intracellular phosphorylated *** Fresh human MSG biopsies were dissociated into single-cell suspensions and permeabilized under optimized conditions. MSG suspensions were stained for cell surface and intracellular markers then analyzed using nine-color conventional flow cytometry, including two intracellular *** Our optimized dissociation and permeabilization protocols for human MSG preserved key cell surface markers. Our flow cytometry panel identified major immune cell populations and distinguished epithelial cells via cytokeratin-18. We demonstrate the protocol's utility showing differential interferon pathway activity in SjD vs. healthy MSG leukocytes and epithelial cells. We provide guidance on panel selection, analytical capabilities, and the impact of cell yield on resolution using conventional flow *** Our optimized protocol enables high-resolution characterization of immune and epithelial cell populations in human MSG, preserving key markers and capturing interferon pathway activity. Our protocol provides a robust framework for the direct study of immune heterogeneity and signaling dynamics in SjD at single cell resolution.

关键词： SC islets   TNF-α   antisense oligonucleotide   cell transplant   inflammation  

摘要： One of the many challenges for islet transplantation as a treatment for type 1 diabetes is inflammation that contributes to islet de-differentiation and death. Innate immune cells such as monocytes and macrophages secrete tumor necrosis factor alpha (TNF-alpha), interleukin 1 beta (IL-1 beta), inducible nitric oxide synthase (iNOS), and IL-6, which directly contribute to islet dysfunction. Attenuation of the early inflammatory response post-transplantation may protect cell survival and subsequent function. Herein, we investigate the development of anti-TNF alpha antisense-oligonucleotide-conjugated polylactide-co-glycolide nanoparticles (PLG-aTNF-alpha NPs) as an anti-inflammatory therapy after stem-cell-derived islet transplantation. PLG-aTNF-alpha NPs are shelf stable and successfully reduce TNF-alpha secretion and expression in inflammatory macrophages. Synergy between the aTNF-alpha antisense oligonucleotide and the polylactide-co-glycolide NPs results in further knockdown of IL-1 beta, IL-6, iNOS, and IL-12 in vitro indicating PLG-aTNF-alpha NPs may protect against the inflammatory cascade in vivo. In a diabetic mouse model, stem-cell-derived islets transplanted to the peritoneal fat were protected after treatment with PLG-aTNF-alpha NPs compared with PLG NPs alone. Tnf alpha and Il1 beta expression was reduced in mice treated with PLG-aTNF-alpha NPs, indicating inflammation was reduced after transplant. PLG-aTNF-alpha NPs reduce TNF-alpha and protect islets, supporting their potential use as a therapeutic in islet transplantation.

关键词： aggregates   IL-6   cadherin   mesenchymal stromal cells   systemic sclerosis  

摘要： Umbilical cord-derived mesenchymal stromal cells (MSCs) transplantation is a promising therapy for systemic sclerosis (SSc) because of their distinctive antifibrotic and immunomodulatory properties. To enhance the effects of MSCs transplantation, novel engineered MSC aggregates preconditioned with human E/N-cadherin fusion protein (hE/N-cad-Fc) and interleukin-6 (IL-6) for SSc treatment are fabricated and named 3D-Cad/IL6-MSCs. These novel-engineered MSC aggregates possess tighter cellular cohesion and exhibit enhanced antiapoptotic, immunosuppressive, and proangiogenic capabilities according to transcriptomic analyses. Moreover, 3D-Cad/IL6-MSCs have improved immunoregulatory effects on peripheral blood mononuclear cells (PBMCs), CD4+ T cells, and CD8+ T cells in vitro because of the synergistic preconditioning from IL-6 and bioactive hE/N-cad-Fc. Upon intravenous injection, 3D-Cad/IL6-MSCs significantly mitigate skin and lung fibrosis and prolong the retention duration in bleomycin (BLM)-induced SSc mouse model. In detail, they suppress excessive infiltration of macrophages and T cells in the injured skin and lungs and reestablish the immune equilibrium of circulating CD4+ T-cell subsets in vivo. These results suggest that 3D-Cad/IL6-MSCs are ideal candidates for SSc therapy and optimize the clinical utilization of MSCs.

关键词： breast neoplasms   interleukin-6   tumor necrosis factor-alpha   prognosis  

摘要： Although cytokines mediate inflammation and inflammation facilitates cancer progression, few studies have evaluated the association between specific cytokines and the prognostic value of breast cancer. Therefore, this study aims to address the following question: What is the prognostic relevance of serum IL-6 and TNF-alpha levels on overall survival and treatment response in women with breast cancer? A systematic review and meta-analysis of cohort studies was conducted. The databases consulted included PubMed/Medline, Web of Science, and EMBASE. A total of 1748 articles were identified, of which 10 were included in the review. A significant association was found between elevated levels of IL-6 and TNF-alpha with poor overall survival and poor treatment response. The meta-analysis showed an HR of 3.74 (95% CI: 1.84-7.6) for elevated IL-6 with high heterogeneity (I2: 61%;p = 0.07) and an HR of 3.13 (95% CI: 1.57-6.23) for TNF-alpha with low heterogeneity (I2: 0%;p = 0.9). The overall response rate was 75% (95% CI: 31-100%;I2: 92%). In conclusion, IL-6 and TNF-alpha emerge as prognostic inflammatory biomarkers in women with breast cancer and are associated with poor survival and poor treatment response. This study highlights the need to establish an international consensus on cutoff points and standardized determination methods to implement these biomarkers in clinical practice.

关键词： LAG3   HIV   NK   IFN-gamma   glycolysis   STAT1  

摘要： Natural killer (NK) cells are integral to the innate immune system and crucial for antiviral defense. NK cell activation and functional state are suppressed by inhibitory receptors. Lymphocyte activation gene 3 (LAG3) is an important inhibitory receptor, but the associated signaling pathways that regulate lymphocyte function remain to be elucidated. In addition, the effect of LAG3 on NK cell function during HIV infection and its specific mechanisms are unclear. In this study, we observed that LAG3 expression by NK cells is elevated in HIV-infected individuals and inversely correlated with CD4/CD8 ratio and CD4+ T cell count. LAG3+ NK cells produce lower levels of interferon-gamma (IFN-gamma), but LAG3-Fc protein significantly enhances NK cell function. The activation of LAG3 significantly inhibits IFN-gamma production and Ki67 expression by NK cells. Our transcriptome sequencing and in vitro data show for the first time that LAG3 not only regulates the transcription of MYC and several glycolysis-related enzyme genes via the PI3K/AKT/mTOR signaling pathway to inhibit glycolysis in NK cells but also suppresses the STAT1/IFNG pathway by upregulating PSAT1 expression, thus limiting IFN-gamma production by NK cells via these two different pathways. Overall, these results provide new insights and identify potential targets for immunotherapy of HIV *** demonstrate that lymphocyte activation gene 3 (LAG3) expression is upregulated on natural killer (NK) cells during HIV infection. LAG3 inhibits glycolysis in NK cells and also upregulates PSAT1 expression to suppress activation of the STAT1/IFNG pathway, thus restricting interferon-gamma production by NK cells. These results provide new clues to study the effects of LAG3 on the metabolism and functional exhaustion of NK cells and offer a potential target for the treatment of HIV.

关键词： ADH   alcohol dehydrogenase   ALDH2   aldehyde dehydrogenase 2   ASCA   anti   Saccharomyces cerevisiae   IgG antibodies   Clec   C-type lectin domain containing   CYP2E1   cytochrome P450 2E1   IL   interleukin   IgA   immunoglobin A   ILC3   innate lymphoid cells type 3   IFNγ   interferon-gamma   iNKT cells   invariant natural killer T cells   LPS   lipopolysaccharide   FMT   fecal microbiota transplantation   FXR   farnesoid X receptor   MAIT cells   mucosa-associated invariant T cells   MAMPs   microbial-associated molecular patterns   MELD   model of end-stage liver disease   Reg3   regenerating islet-derived protein 3   TNF   tumor necrosis factor   TNFR1   TNF receptor 1   TLR   Toll-like receptor  

摘要： Alcohol-associated organ damage is a major cause of morbidity and mortality worldwide, with the liver being the primarily affected organ. Emerging evidence highlights the gut microbiota as a key driver in alcohol-associated liver disease. Changes in the microbiota composition, microbial translocation, and a dysregulated immune response culminate in inflammation and tissue injury. Persistent liver injury eventually promotes disease progression from steatohepatitis to fibrosis and cirrhosis. In this review, we provide an overview of microbiota alterations observed in patients with chronic alcohol consumption and explore the mechanisms by which microbiota contributes to alcohol-associated damage across various organ systems. Emphasis is placed on changes in the gut microbiota and its role in alcohol-associated liver disease, the disease condition with the most robust evidence linking microbiota-related changes to organ injury. Additionally, we highlight key areas where further research is needed to address unresolved questions. Finally, we discuss emerging therapeutic strategies targeting the microbiota to treat alcohol use disorders and prevent alcohol-associated liver diseases.

关键词： T. pyogenes   PLO   NF-kappa B   NLRP3   caspase-1   IL-1 beta  

摘要： Trueperella pyogenes (T. pyogenes) is an important opportunistic pathogen in animals and can also cause diseases in humans. Previous studies have shown that T. pyogenes infection can upregulate the levels of pro-inflammatory cytokines, such as interleukin-1 beta (IL-1 beta), in host tissues. However, the underlying mechanisms are not yet fully understood. In the current study, we found that both inactivated T. pyogenes cells (iTP) and pyolysin (PLO, a major virulence factor of T. pyogenes) can promote the transcription of the IL-1 beta gene both in vivo and in vitro. iTP-caused upregulation of IL-1 beta gene transcription is dependent on nuclear factor-kappa B (NF-kappa B). On the other hand, we determined that PLO, but not iTP, can promote the maturation of IL-1 beta by activating caspase-1-mediated processing of pro-IL-1 beta. Further, we confirmed that PLO can induce potassium ion (K+) efflux in mouse macrophages, thereby activating caspase-1 in a Nod-like receptor protein 3 (NLRP3)-dependent manner. Blocking K+ efflux or knocking down the expression of NLRP3 both inhibited caspase-1 activation and pro-IL-1 beta processing. Taken together, these findings demonstrate that T. pyogenes can promote IL-1 beta expression at both the transcriptional and post-translational levels in a murine macrophage model. These results significantly enhance our understanding of the pathogenesis of T. pyogenes and the interactions between T. pyogenes and host immune system.