Gasdermin D in macrophages drives orchitis by regulating inflammation and antigen presentation processes.

Inflammation in the testes induced by infection and autoimmunity contributes significantly to male infertility, a public health issue. Current therapies using antibiotics and broad-spectrum anti-inflammatory drugs are ineffective against non-bacterial orchitis and induce side effects. This highlights the need to explore the pathogenesis of orchitis and develop alternative therapeutic strategies. In this study, we demonstrated that Gasdermin D (GSDMD) was activated in the testes during uropathogenic Escherichia coli (UPEC)-induced acute orchitis, and that GSDMD in macrophages induced inflammation and affected spermatogenesis during acute and chronic orchitis. In testicular macrophages, GSDMD promoted inflammation and antigen presentation, thereby enhancing the T-cell response after orchitis. Furthermore, the pharmacological inhibition of GSDMD alleviated the symptoms of UPEC-induced acute orchitis. Collectively, these findings provide the first demonstration of GSDMD's role in driving orchitis and suggest that GSDMD may be a potential therapeutic target for treating orchitis.

Serum human epididymis secretory protein 4 correlates with sepsis-associated acute respiratory distress syndrome and 28-day mortality in critically ill patients.

BACKGROUND: Acute respiratory distress syndrome (ARDS) is a severe disease with high mortality, and its primary cause is sepsis. The aim of this study was to detect and evaluate the role of Human epididymis protein 4 (HE4) in sepsis-related ARDS. METHODS: One hundred and twenty-three critically ill sepsis patients with/without ARDS and 102 healthy controls were enrolled in this study. Blood samples were collected upon admission for quantitative testing of HE4 by chemiluminescent microparticle immunoassay (CMIA). ROC curve analysis and Spearman's correlation analysis were conducted to determine the diagnostic and prognostic value of HE4. RESULTS: Compared with controls, the serum HE4 concentrations of sepsis patients were elevated, and levels in sepsis patients with ARDS were significantly higher (all p < 0.0001). Moreover, HE4 concentrations were strongly correlated with the clinical severity characteristics of sepsis patients, and ROC curve suggested that the AUC of HE4 applied to discriminate sepsis-ARDS patients from sepsis patients was 0.903. HE4 was also found to be a prognostic biomarker of clinical severity and 28-day mortality among critically ill sepsis patients. Logistic regression analysis showed that HE4 was an independent factor for diagnosis of ARDS. Meanwhile, ROC curve analysis showed that the cut-off value of serum HE4 to discriminate 28-day mortality from sepsis patients (AUC: 0.782) was 646.5 pmol/L. CONCLUSIONS: The concentration of serum HE4 in patients with sepsis-related ARDS was markedly increased and was significantly correlated with mortality, which suggests that serum HE4 could be a promising diagnostic and prognostic biomarker for ARDS in sepsis patients.

[Optimizing the methods of whole lung cancer RNA-loaded dendritic cells].

BACKGROUND: Dendritic cells (DCs) are the unique antigen-presenting cells that can activate naive T lymphocytes. This function is critical for inducing specific immune response. DCs-based vaccines have been used broadly in immunotherapy for many carcinomas. Constructing vaccines by transfecting total tumor RNA into DCs can be done with a few tumor tissues and need not to identify tumor antigens, so it is especially suitable for lung cancer which lacks tumor-specific antigens but has great heterogenicity and weak immunogenicity. Currently, the best transfection stage and method are still indefinite. So, the objective of this study is to explore the best condition of transfecting total RNA extracted from lung cancer tissues into DCs. METHODS: Ten patients with lung cancer were enrolled whose tumor tissues were CEA and MUC1 positive in immunohistochemical staining. Total tumor RNA were extracted by one-step method. Then DCs and T cells were separated and cultured from peripheral blood monocytes and the RNA was transfected into the DCs in different stages with different methods. CEA and MUC1 expression in the transfected DCs were measured by flow cytometry analysis and T cells' proliferation was examined by mixed lymphocyte reaction (MLR). RESULTS: The expression of CEA and MUC1 protein in immature DCs (11.33±2.64, 39.68±7.25) was remarkably higher than that in mature DCs (5.46±1.63, 27.17±4.16) after transfection with total RNA of lung cancer tissues (P < 0.01), and the DCs presented more powerful effects on T cell proliferation. The CEA and MUC1 expression on DCs were significantly higher in electroporation transfection group (20.53±3.64, 65.39± 9.33) than that in lipofection group (11.33±2.64, 39.68±7.25) and passive pulsing transfection group ( 0.91±0.27,18.53±3.26)(P < 0.01), and the DCs in electroporation transfection group presented more powerful effects on stimulating T cell proliferation than the other two groups did. CONCLUSIONS: Transfecting total tumor RNA into immature DCs by using electroporation is a good way to construct DCs-based vaccines for lung cancer and to achieve a higher activity to stimulate T cell proliferation.