Jianzhang Hu

The mechanisms underlying cellular damage induced by the mitochondrially encoded NADH dehydrogenase subunit 4 (MT-ND4) with R340H mutation caused by the variant m.11778G > A in complex I are intricate. Numerous studies indicate that mitochondria play a primary role in cellular death due to this mutation. However, the detailed pathological effects remain incompletely elucidated. To decipher the specific impacts of this mutation on cellular death, mitochondrial dysfunction was investigated in 661 W cells expressing exogenous Mut-ND4 (m.G11778A). Importantly, the oxygen consumption rate (OCR) assessed by Seahorse XF analyzer exhibited a significant decrease under galactose conditions and an excessive production of reactive oxygen species (ROS). Conversely, the activity levels of catalase (CAT), superoxide dismutase (SOD), and glutathione disulfide (GSSG) were decreased, leading to increased cell death in cells expressing Mut-ND4 (m.G11778A) under galactose conditions. In addition, structural disruptions in the optic nerves of mice subjected to Mut-ND4-AAV infection were revealed. These findings suggest that Mut-ND4 (m.G11778A) contributes to cellular injury and an oxidative stress imbalance, characterized by decreased mitochondrial oxygen consumption, increased oxidative products, and reduced antioxidant capacity.

Traditional clinical medical internship models often lack structured educational objectives and comprehensive assessments, limiting holistic competency development. This study evaluates the effectiveness of a novel internship education management model integrating Bloom's Taxonomy with a student-mentor rotation system to enhance cognitive, affective, and psychomotor skills. We conducted a randomized controlled trial with 54 optometry interns during their ophthalmology clinical rotation, assigning them to either an experimental group ( n = 30) using the Bloom's Taxonomy-integrated rotation model or a control group ( n = 24) receiving traditional training. Both groups completed 12-week rotations with identical duration and content. The Bloom's Taxonomy-integrated rotation model was organized into three progressive phases corresponding to ascending levels of Bloom's Taxonomy (knowledge acquisition → clinical application → synthesis). Outcomes were assessed through written examinations, practical skill test, and course experience questionnaires. The Bloom's Taxonomy-integrated rotation group showed superior performance in case analysis, history taking and practical skills. They also reported significantly higher satisfaction in problem-solving, motivation and feedback quality. In conclusion, the Bloom's Taxonomy-integrated rotation model demonstrated superior clinical reasoning, practical skills, and medical interns' satisfaction, among optometry interns compared to traditional training. The Bloom's Taxonomy-integrated rotation model represents an effective, structured framework for enhancing medical internship training, though warranting further validation in broader settings.

Background: Mounting evidence indicates metabolic dysregulation in diabetic corneal neuropathy (DCN). This study elucidates how the chromatin remodeler Brahma-related gene 1 (BRG1) orchestrates glycolytic reprogramming to drive neurodegeneration and epithelial repair defects in DCN. Methods: Type 1 diabetic mice were established via streptozotocin (STZ) injection. Glycolysis was inhibited using 2-deoxy-D-glucose (2-DG) to assess its role in DCN pathogenesis. BRG1 expression was modulated by subconjunctival plasmid delivery (overexpression/knockdown). Pathway screening identified BRG1 downstream effectors, and phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT) inhibition (LY294002) confirmed regulatory hierarchy. Glycolytic flux was evaluated via Western blotting and immunofluorescence; corneal nerve integrity and epithelial healing were assessed by βIII-tubulin staining and sodium fluorescein assay. Results: Hyperglycemia upregulated BRG1 and glycolytic enzymes in diabetic corneal nerves. BRG1 overexpression exacerbated epithelial repair delay and neurodegeneration, while knockdown partially reversed damage. BRG1 overexpression activated PI3K/AKT transcription, and PI3K/AKT inhibition did not alter BRG1 levels but rescued BRG1-induced pathologies. Conclusions: Glycolytic reprogramming is a critical driver of DCN progression. BRG1 activates PI3K/AKT signaling to enhance glycolytic flux, thereby regulating DCN pathogenesis. Targeting this axis may offer novel therapeutic strategies.

Objective: To investigate the anti-inflammatory effect of glycolysis rate-limiting enzyme 6-phosphofructose-2-kinase/fructose-2, 6-biphosphatase 3 (PFKFB3) in fungal keratitis (FK) infected by Fusarium solani ( F. solani ). Methods: We identified the up-regulation of PFKFB3 in fungal keratitis via western blot, quantitative real-time polymerase chain reaction (RT-PCR), and immunofluorescence staining. Subsequently, elucidated the augmentation of glycolytic flux in cornea and bone marrow-derived macrophages (BMDM) following F. solani invasion by RT-PCR, cellular energy metabolism analyzer, and lactate content assay. After that, we reduced PFKFB3 expression utilizing small interfering RNA (siRNA) in vitro and adeno-associated virus (AAV) in vivo and also assessed the expression levels of inflammatory factors. The severity of corneal infection following PFKFB3 depletion was checked by slit-lamp microscopy, corneal OCT, and H&E staining. Ultimately, we assessed the phosphorylation status of the PI3K/AKT/NF-κB p65 signaling pathway following PFKFB3 suppression via western blot and immunofluorescence staining. Results: PFKFB3 was highly triggered in F. solani -infected corneas and BMDM compared to normal tissue. Besides, infection with F. solani promotes the increase of inflammatory mediators and glycolytic flux in the cornea and BMDM. Whereas inflammation in BMDM and the degree of fungal keratitis lesions worsen by suppressing PFKFB3 expression, which increased corneal ulcer infiltration, elevated clinical scores, enhanced corneal thickness, and upregulation of inflammatory signals could be demonstrated. Furthermore, we found that F. solani infection can activate the phosphorylation of PI3K/AKT/NF-κB p65 at low PFKFB3 expression levels. Conclusions: In F. solani -infected corneas and BMDM, the glycolysis rate-limiting enzyme PFKFB3 was markedly upregulated. After infection, moderate PFKFB3 activation effectively mitigates inflammation and the progression of fungal keratitis. Moreover, activated PFKFB3 may rely on the PI3K/AKT/NF-κB p65 signaling pathway to safeguard the cornea from further damage due to inflammation.

Objectives This study aims to develop a deep learning algorithm (DLA) using the InceptionV3 architecture for effective diabetic peripheral neuropathy (DPN) screening via corneal confocal microscopy (CCM) images. Design Retrospective study. Setting Ophthalmology Centre of General Hospital. Participants 127 participants enrolled: 33 healthy participants, 57 diabetic patients with DPN (DPN+) and 37 diabetic patients without DPN (DPN−). Interventions Not applicable. Main outcome measures The CCM image dataset, which was collected from participants (with five images per eye), was randomly divided into training, validation and test subsets in a 7:1:2 ratio. The images were preprocessed, augmented and used to train the InceptionV3 model. We compared its performance against the ResNet, DenseNet and Swin Transformer models. Performance was evaluated using accuracy, recall, F1 score and area under the curve (AUC) metrics. Results For single-participant predictions, the InceptionV3 model achieved the highest accuracy (0.9231), recall (0.8846), F1 score (0.9020) and AUC (0.9534) compared with the other models. For single-image predictions in the three-class classification task of CCM images, the InceptionV3 model achieved a precision of 0.8385, a recall of 0.9083, an F1 score of 0.8720 and an AUC of 0.8769 for predicting DPN+. Conclusions The InceptionV3-based DLA model achieved superior performance compared with traditional convolutional neural network architectures like ResNet and DenseNet, and the Swin transformer model, highlighting its potential for effective DPN screening.

Background: Diabetic keratopathy (DK) is a common ocular complication of diabetes, with its progression closely linked to autophagy regulation. This study aims to explore the role of long non-coding RNAs (lncRNAs) in modulating autophagy during diabetic pathogenesis, focusing on lncRNA general transcription factor IIIC subunit 1 (GTF3C1) and its potential as a therapeutic target for diabetic corneal neuropathy (DCN). Methods: High-throughput sequencing identified dysregulated lncRNAs in the trigeminal ganglia of diabetic mice. Functional validation included mechanistic studies on lncRNA GTF3C1, miR-542-3p, and autophagy-related targets. Autophagy activity, corneal nerve density, and epithelial healing were quantified using quantitative real-time polymerase chain reaction (qRT-PCR), immunofluorescence, and histology in diabetic models. Results: lncRNA GTF3C1 was significantly downregulated in diabetic trigeminal ganglion (TG). It functioned as a molecular sponge for miR-542-3p, alleviating its repression on GABA type A receptor-associated protein (GABARAP) and phosphatase and tensin homolog (PTEN), thereby enhancing autophagy activity. This process promoted corneal nerve fiber regeneration and epithelial wound healing in diabetic mice. Conclusions: Our findings highlight lncRNA GTF3C1 as a critical regulator of autophagy in diabetic corneal nerves, offering a potential diagnostic and therapeutic target for DCN. This study provides molecular insights into the pathogenesis of DCN and lays the groundwork for future clinical strategies.

Fungal keratitis (FK) is a challenging and sight-threatening corneal disease caused by fungal infections. Although long noncoding RNAs (lncRNAs) have been explored in various infectious diseases, their specific roles in FK remain largely unexplored. A mouse model of FK was created by infecting corneal stromal cells with Fusarium solani. High-throughput lncRNA expression profiling was conducted on FK-affected corneal tissues to identify differentially expressed lncRNAs. Reverse transcription quantitative PCR (RT-qPCR) was used to validate the results. A competing endogenous RNA (ceRNA) network was constructed. Additionally, a specific antisense oligonucleotide (ASO) targeting lncRNA ENSMUST00000226838/Osr2 (Osr2) was developed for therapeutic evaluation. Inflammatory markers IL-1β, IL-6, and TNF-α were measured, and corneal inflammation was assessed through histological analysis and slit-lamp examination. Fluorescent in situ hybridization (FISH) was used to confirm Osr2 localization, whereas a dual-luciferase reporter assay verified interactions between Osr2 and miR-30a-3p. We identified 1143 differentially expressed lncRNAs in FK, with 701 upregulated and 442 downregulated. The ceRNA network analysis indicated that lncRNA Osr2 regulates Xcr1 expression through miR-30a-3p. Treatment with ASO-Osr2 significantly reduced corneal inflammation, and FISH confirmed lncRNA Osr2 distribution in both the nucleus and cytoplasm. Dual-luciferase assays demonstrated the interaction between Osr2 and miR-30a-3p, highlighting their potential roles in the progression of FK. This study outlined the lncRNA expression profile in FK and established a ceRNA regulatory network, identifying lncRNA Osr2 as a crucial modulator of FK pathogenesis through its interaction with miR-30a-3p. These findings highlighted lncRNA Osr2 as a promising therapeutic target for the treatment of FK.