Objective:To evaluate the effect of mushroom-like pattern femtosecond laser enabled penetrating keratoplasty with different laser energies on the ring incision and corneal endothelial cells, and compare with conventional penetrating keratoplasty.Methods:According to the point/line separation and blasting energy, 48 porcine corneas were randomly divided into 6 groups by using a random number table.In group A, B, C, D and E, the point/line separation was 4/4, 4/4, 8/8, 8/8 and 4/2 μm, and the blasting energy was 1.5, 2.0, 1.5, 2.0 and 2.0 μJ; In group F, a negative pressure ring drill was used to make penetrating incisions.There were 8 porcine corneas in each group.Femtosecond laser of 200 kHz was used to make corresponding mushroom-shaped penetrating incisions on the porcine corneas, and compared with porcine corneas incisions with a ring drill.The optical and scanning electron microscope images were used to evaluate the glossiness of ring incisions.Four human corneas used femtosecond laser for mushroom-like penetrating incisions with point/line separation 8/8 μm, and blasting energy 2.0 μJ as the experimental group; three human corneas received a negative pressure ring drill for penetrating incisions as the control group.The loss rate of corneal endothelial cells was observed and compared between experimental group and control group.The donor guardians agreed to the acquisition and use of the donors' cornea and signed informed consents.This study protocol was approved by the Ethics Committee of Nanjing First Hospital.Results:Femtosecond laser mushroom penetrating ring incision completion rate was 100%.The corneal ring incision in each group was stepped, and sections of ring incisions in group A and group E were glossiest by the light microscope.Sections of ring incisions in group E were glossiest by the laser scanning electron microscope.The overall glossiness scores of the corneal ring incisions among various groups were statistically significantly different by the light microscopy ( F=22.75, P<0.01). Among them, the glossiness scores in the group A were higher than those in the group B, and the glossiness scores in the group C were higher than those in the group D, with statistically significant differences (both P<0.05). The overall glossiness scores of corneal ring incisions among various groups were statistically significantly different by laser scanning electron microscopey ( F=122.33, P<0.01). Among them, the glossiness scores in the group A were higher than those in the group B, and the the glossiness scores in the group C were higher than those in the group D, with statistically significant differences (both P<0.05). The corneal endothelial cells showed regular shape and tight arrangement in the experimental group, and irregular shape and loose connections were seen in the control group.The average corneal endothelial cell loss rate in the experimental group was (2.2±1.3)%, lower than (6.7±2.1)% of the control group, with a significant difference between them ( t=3.569, P<0.05). Conclusions:Femtosecond laser can produce perfect mushroom configuration, and the ring incision glossiness is better in comparison with trephine cutting.Femtosecond laser ring cut can lessen corneal endothelial loss.

Inhalation of crystalline silicon dioxide particles can induce silicosis, and the development of silicosis is closely related to the occurrence and development of pulmonary inflammation and pulmonary fibrosis. NOD-like receptor thermal protein domain associated protein 3 (NLRP3) inflammasome has been established as a major proinflammatory receptor for sensing environmental danger signals. Activation of NLRP3 inflammasomes after phagocytosis of silicon dioxide particles by pulmonary macrophages may be an important mechanism to induce oxidative stress and sustained inflammatory response in the lung. This article summarizes the role of NLRP3 inflammasome in the inflammatory response and pulmonary fibrosis in silicosis, and analyzes it as a potential target for silicosis treatment.

Inhalation of crystalline silicon dioxide particles can induce silicosis, and the development of silicosis is closely related to the occurrence and development of pulmonary inflammation and pulmonary fibrosis. NOD-like receptor thermal protein domain associated protein 3 (NLRP3) inflammasome has been established as a major proinflammatory receptor for sensing environmental danger signals. Activation of NLRP3 inflammasomes after phagocytosis of silicon dioxide particles by pulmonary macrophages may be an important mechanism to induce oxidative stress and sustained inflammatory response in the lung. This article summarizes the role of NLRP3 inflammasome in the inflammatory response and pulmonary fibrosis in silicosis, and analyzes it as a potential target for silicosis treatment.

Despite complications were significantly reduced due to the popularity of percutaneous coronary intervention (PCI) in clinical trials, reperfusion injury and chronic cardiac remodeling significantly contribute to poor prognosis and rehabilitation in AMI patients. We revealed the effects of HSP47 on myocardial ischemia-reperfusion injury (IRI) and shed light on the underlying molecular mechanism. We generated adult mice with lentivirus-mediated or miRNA (mi1/133TS)-aided cardiac fibroblast-selective HSP47 overexpression. Myocardial IRI was induced by 45-min occlusion of the left anterior descending (LAD) artery followed by 24 h reperfusion in mice, while ischemia-mediated cardiac remodeling was induced by four weeks of reperfusion. Also, the role of HSP47 in fibrogenesis was evaluated in cardiac fibroblasts following hypoxia-reoxygenation (HR). Extensive HSP47 was observed in murine infarcted hearts, human ischemic hearts, and cardiac fibroblasts and accelerated oxidative stress and apoptosis after myocardial IRI. Cardiac fibroblast-selective HSP47 overexpression exacerbated cardiac dysfunction caused by chronic myocardial IRI and presented deteriorative fibrosis and cell proliferation. HSP47 upregulation in cardiac fibroblasts promoted TGFβ1-Smad4 pathway activation and Smad4 deubiquitination by recruiting ubiquitin-specific peptidase 10 (USP10) in fibroblasts. However, cardiac fibroblast specific USP10 deficiency abolished HSP47-mediated fibrogenesis in hearts. Moreover, blockage of HSP47 with Col003 disturbed fibrogenesis in fibroblasts following HR. Altogether, cardiac fibroblast HSP47 aggravates fibrosis post-myocardial IRI by enhancing USP10-dependent Smad4 deubiquitination, which provided a potential strategy for myocardial IRI and cardiac remodeling.

High mobility group box 1 (HMGB1) has been reported to play an important role in experimental autoimmune encephalomyelitis (EAE). Astrocytes are important components of neurovascular units and tightly appose the endothelial cells of microvessels by their perivascular endfeet and directly regulate the functions of the blood-brain barrier. Astrocytes express more HMGB1 during EAE while the exact roles of astrocytic HMGB1 in EAE have not been well elucidated. Here, using conditional-knockout mice, we found that astrocytic HMGB1 depletion decreased morbidity, delayed the onset time, and reduced the disease score and demyelination of EAE. Meanwhile, there were fewer immune cells, especially pathogenic T cells infiltration in the central nervous system of astrocytic HMGB1 conditional-knockout EAE mice, accompanied by up-regulated expression of the tight-junction protein Claudin5 and down-regulated expression of the cell adhesion molecules ICAM1 and VCAM1 in vivo. In vitro, HMGB1 released from astrocytes decreased Claudin5 while increased ICAM1 and VCAM1 expressed by brain microvascular endothelial cells (BMECs) through TLR4 or RAGE. Taken together, our results demonstrate that HMGB1 derived from astrocytes aggravates EAE by directly influencing the immune cell infiltration-associated functions of BMECs.
Mice ; Animals ; Encephalomyelitis, Autoimmune, Experimental/metabolism* ; Astrocytes/metabolism* ; HMGB1 Protein/metabolism* ; Endothelial Cells/metabolism* ; Mice, Inbred C57BL ; Mice, Knockout ; Blood-Brain Barrier/metabolism*