CCR5-overexpressing mesenchymal stem cells protect against experimental autoimmune uveitis: insights from single-cell transcriptome analysis.

Autoimmune uveitis is a leading cause of severe vision loss, and animal models provide unique opportunities for studying its pathogenesis and therapeutic strategies. Here we employ scRNA-seq, RNA-seq and various molecular and cellular approaches to characterize mouse models of classical experimental autoimmune uveitis (EAU), revealing that EAU causes broad retinal neuron degeneration and marker downregulation, and that Müller glia may act as antigen-presenting cells. Moreover, EAU immune response is primarily driven by Th1 cells, and results in dramatic upregulation of CC chemokines, especially CCL5, in the EAU retina. Accordingly, overexpression of CCR5, a CCL5 receptor, in mesenchymal stem cells (MSCs) enhances their homing capacity and improves their immunomodulatory outcomes in preventing EAU, by reducing infiltrating T cells and activated microglia and suppressing Nlrp3 inflammasome activation. Taken together, our data not only provide valuable insights into the molecular characteristics of EAU but also open an avenue for innovative MSC-based therapy.

Smurf1 Modulates Smad Signaling Pathway in Fibrotic Cataract Formation.

Purpose: TGF-ß/BMP signaling pathway plays a significant role in fibrotic cataract. Smurf1, a ubiquitin protein ligase, regulates the TGF-ß/BMP signaling pathway through the ubiquitin-proteasome system (UPS). This study aims to investigate the role of Smurf1 in the progression of fibrotic cataract and its underlying mechanism. Methods: We used a mouse model of injury-induced anterior subcapsular cataract (ASC) and administered the Smurf1 inhibitor A01 for in vivo investigations. RNA sequencing was performed to examine global gene expression changes. Protein levels were assessed by Simple Western analysis. The volume of subcapsular opacity was determined using whole-mount immunofluorescence of lens anterior capsules. Lentivirus was utilized to establish cell lines with Smurf1 knockdown or overexpression in SRA01/04. Lens epithelial cell (LEC) proliferation was evaluated by CCK8 and EdU assays. Cell cycle profile was determined by flow cytometry. LEC migration was measured using Transwell and wound healing assays. Results: The mRNA levels of genes associated with cell proliferation, migration, epithelial-mesenchymal transition (EMT), TGF-ß/BMP pathway, and UPS were upregulated in mouse ASC model. Smurf1 mRNA and protein levels were upregulated in lens capsules of patients and mice with ASC. Anterior chamber injection of A01 inhibited ASC formation and EMT. In vitro, Smurf1 knockdown reduced proliferation, migration and TGF-ß2-induced EMT of LECs, concomitant with the upregulation of Smad1, Smad5, and pSmad1/5. Conversely, overexpression of Smurf1 showed opposite phenotypes. Conclusions: Smurf1 regulates fibrotic cataract progression by influencing LEC proliferation, migration, and EMT through the modulation of the Smad signaling pathway, offering a novel target for the fibrotic cataract treatment.

BMP-4 and BMP-7 Inhibit EMT in a Model of Anterior Subcapsular Cataract in Part by Regulating the Notch Signaling Pathway.

Purpose: The proliferation, migration, and epithelial-mesenchymal transition (EMT) of lens epithelial cells (LECs) are believed to be the pathological mechanisms underlying anterior subcapsular cataract (ASC). Bone morphogenetic proteins (BMPs) inhibit transforming growth factor-beta (TGF-ß)-induced fibrosis in the lens. Herein, we aimed to further clarify the roles of BMP-4/BMP-7 in the progression and the underlying mechanisms of fibrotic cataract. Methods: BMP-4/BMP-7, TGF-ß2, jagged-1 peptide, or DAPT were applied in a mouse injury-induced ASC model and in the human LEC cell line SRA01/04. The volume of opacity was examined by a slit lamp and determined by lens anterior capsule whole-mount immunofluorescence. Global gene expression changes were assessed by RNA sequencing, and the levels of individual mRNAs were validated by real-time PCR. Protein expression was determined by the Simple Western sample dilution buffer. Cell proliferation was examined by CCK8 and EdU assays, and cell migration was measured by Transwell and wound healing assays. Results: Anterior chamber injection of BMP-4/BMP-7 significantly suppressed subcapsular opacification formation. RNA sequencing of the mouse ASC model identified the Notch pathway as a potential mechanism involved in BMP-mediated inhibition of ASC. Consistently, BMP-4/BMP-7 selectively suppressed Notch1 and Notch3 and their downstream genes, including Hes and Hey. BMP-4/BMP-7 or DAPT suppressed cell proliferation by inducing G1 cell cycle arrest. BMP-4/BMP-7 also inhibited TGF-ß2-induced cell migration and EMT by modulating the Notch pathway. Conclusions: BMP-4/BMP-7 attenuated ASC by inhibiting proliferation, migration, and EMT of LECs via modulation of the Notch pathway, thereby providing a new avenue for ASC treatment.

Field measurement of local wind environment on the approach deck of a suspension bridge in mountain terrain.

The local wind environment above the bridge deck affects the aerodynamic characteristics of vehicles, thus affecting the driving safety of the bridge deck. Influenced by the mountain topography and the bridge deck's ancillary facilities, the local wind environment above the bridge deck is complex and changeable, and its impact on the bridge deck traffic can not be ignored. In order to accurately evaluate the local wind field parameters, a monitoring system of the local wind environment is developed. Utilizing the monitoring system, wind parameters above the approach deck of a long-span suspension bridge in a mountain area are measured. The relationship of wind characteristics between the incoming flow and the wind above the bridge deck is investigated. Results show the significant difference between the local wind environment above the bridge deck and the incoming flow's characteristics; the wind profile above the bridge deck does not follow the exponential distribution; the equivalent height of the wind load on the vehicle is higher than the vehicle's gravity centre. This study is relevant for studying the local wind environment, driving safety, and serviceability of long-span bridges in mountainous areas.

Playing for a Resilient Future: A Serious Game Designed to Explore and Understand the Complexity of the Interaction among Climate Change, Disaster Risk, and Urban Development.

Urban development and disaster risk are deeply linked, especially now when we are facing increasingly frequent climate change. Hence, knowledge of the potential trade-offs between urban development and disaster risk reduction (DRR) may have potential to build a resilient and sustainable future. The objectives of this study are (1) to present education for a sustainability (EfS) program and to evaluate its performance: a serious game of knowledge communication for the interactions among climate change, disaster risk, and urban development; (2) to explore factors that will influence the players' decision making in the trade-offs between urban development and DRR under an urbanization background through counterfactual scenarios constructed by a series of serious games. The Yudai Trench, once a critical component of the urban green infrastructure of ancient Guangzhou, has disappeared under rapid urban expansion, leaving the city exposed to environmental hazards caused by climate change. Is the disappearance of the Yudai Trench an inevitable event in the progress of urbanization? To answer this question, the study constructed counterfactual scenarios by recuring the historical progress through the same serious game. Gameplay involved the players' decision making with associated impacts on the urbanization progress and the DRR in diverse climate hazard scenarios. For this study, 107 undergraduates from related majors, who are also would-be policymakers, were selected as players. The methodology combined questionnaire survey and participant observation complemented by interviews. The t-test results indicated that undergraduates' knowledge levels had significant positive changes after the end of the serious game. Importantly, the results showed that the knowledge could potentially contribute to the players' decision-making process for DRR by assisting them in making pre-decision. Beside this knowledge, the results expanded the range of influencing factors and solutions reported by previous literature on DRR under an urbanization background against climate hazards by constructing counterfactual scenarios, e.g., higher economic levels and policy incentives. In this study, the serious game was evaluated as an innovative communication and the EfS method in counterfactual scenarios. These findings of the study provide a reference for future practice, policymaking, and decision making so as to help harness lessons learned from unrealized environmental hazards to support a more resilient future through informed policies and plans.