OBJECTIVE:Physcion, a natural anthraquinone, possesses antifungal and anti-inflammatory activities. This study aimed to investigate its antifungal and anti-inflammatory effects in a murine model of Aspergillus fumigatus (AF) keratitis (AFK). METHODS:The safety of physcion was evaluated using Cell Counting Kit-8 assays, live/dead cell staining and the Draize test. Antifungal activity was assessed by propidium iodide staining, calcofluor white staining, crystal violet biofilm assays and determination of the MIC. Network pharmacology analysis was performed to predict potential therapeutic targets of physcion. Both in vitro and in vivo experiments were conducted using AF spore-infected mouse corneas and RAW264.7 macrophages treated with physcion or phosphate-buffered saline. Inflammatory responses were evaluated by slit-lamp examination, immunofluorescence, real-time quantitative polymerase chain reaction (RT-qPCR) and western blotting (WB), focusing on neutrophil and macrophage recruitment together with the expression of pro-inflammatory proteins and components of the NF-κB signalling pathway. RESULTS:Physcion effectively inhibited the growth of AF. In vivo, physcion-treated mice exhibited significantly lower clinical scores compared with untreated infected controls. Immunofluorescence analysis revealed markedly decreased infiltration of neutrophils/macrophages in the corneas of physcion-treated mice. RT-qPCR together with WB analyses demonstrated that physcion treatment obviously reduced the phosphorylated NF-κB p65 and phosphorylated IκB, increased IκB expression and suppressed the expression of IL-1β, TNF-α and IL-18 in infected mouse corneas and RAW264.7 cells. Consistently, IF imaging of RAW264.7 cells showed reduced nuclear localization of NF-κB p65 following physcion treatment. CONCLUSION:Physcion alleviated AFK by inhibiting fungal growth, limiting neutrophil and macrophage recruitment, and suppressing activation of the NF-κB signalling pathway.

The critical gap between clinically required durability and metabolic functionality in corneal prostheses drives the need for biomaterials that recapitulate the structural hierarchy of cornea. To address this, we introduce a solvent-free water enabling the self-assembly of dodecenyl-modified agarose (DAA) into hydrogels that recapitulate the structural hierarchy of native cornea. The DAA hydrogel exhibits a microporous architecture with pore sizes analogous to the corneal stroma, facilitating essential mass transport while maintaining ultraviolet (UV)-protective transparency (> 90% visible light transmittance). Unlike brittle and chemically crosslinked agarose hydrogels, the DAA hydrogel achieves tissue-like mechanical properties, including high tensile strength of 1.66 MPa and suture retention strength of 1.57 N, due to the synergistic effects of hydrophobic microdomains and hydrogen bonding between DAA chains. In addition, the wVIPS process eliminates toxic crosslinkers, ensuring 180 days stability in artificial tears and excellent biocompatibility. Consequently, the DAA hydrogel demonstrated seamless host integration without scarring and promoted restoration of corneal barrier function in a rabbit lamellar keratoplasty model. By establishing microphase separation as a universal design strategy, this work advances polysaccharide-based biomaterials for loadbearing ophthalmological applications, effectively bridging nanoscale self-assembly with macroscale tissue functionality.

To evaluate the efficacy of selective laser trabeculoplasty (SLT) on intraocular pressure (IOP) for the normal tension glaucoma (NTG) in Chinese. Consecutive patients with NTG treated with monocular SLT for eye with higher IOP were enrolled. IOP was evaluated for each patient before SLT, 1 h, 1 week, 1, 3, 6 months, 1 and 2 years after treatment. The treatment success was defined as IOP reduction from the preoperative IOP by 20

Purpose Myosin 1f (Myo1f), an unconventional long-tailed class Ⅰ myosin, plays significant roles in immune cell motility and innate antifungal immunity. This study was aimed to assess the expression and role of Myo1f in Aspergillus fumigatus (AF) keratitis. Methods Myo1f expression in the corneas of mice afflicted with AF keratitis and in AF keratitis-related cells was assessed using protein mass spectrometry, quantitative real-time polymerase chain reaction (qRT-PCR), western blotting, and immunofluorescence. Myo1f expression following pre-treatment with inhibitors of dendritic cell-associated C-type lectin-1 (Dectin-1), Toll-like receptor 4 (TLR-4), and lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) was also examined. In AF keratitis mouse models, Myo1f small interfering RNA (siRNA) was administered via subconjunctival injection to observe disease progression, inflammatory cell recruitment, and protein production using slit lamp examination, immunofluorescence, hematoxylin-eosin (HE) staining, and western blotting. Results Myo1f expression was upregulated in both AF keratitis mouse models and AF keratitis-related cells. Dectin-1, TLR-4, and LOX-1 were found to be essential for the production of Myo1f in response to the infection with AF. In mice with AF keratitis, knockdown of Myo1f reduced disease severity, decreased the recruitment of neutrophils alongside macrophages to inflammatory areas, suppressed the myeloid differentiation factor 88 (MyD88)/ nuclear factor-kappaB (NF-κB) signaling pathway, and decreased the production of interleukin (IL)-1β, tumor necrosis factor (TNF)-α, along with IL-6. Additionally, Myo1f was associated with apoptosis and pyroptosis in mice with AF keratitis. Conclusions These findings demonstrated that Myo1f contributed to the recruitment of neutrophils and macrophages, the production of pro-inflammatory cytokines, and was associated with apoptosis and pyroptosis during AF keratitis.

AIM To explore whether CD3u03B5 is involved in the adaptive immunity of Aspergillus fumigatus (A. fumigatus) keratitis in mice and the role of innate and adaptive immunity in it. METHODS Mice models of A. fumigatus keratitis were established by intra-stromal injection and corneal epithelial scratching. Subconjunctival injections of natamycin, wedelolactone, LOX-1 inhibitor (poly u2160) or Dectin-1 inhibitor (laminarin) were used to treat mice with A. fumigatus keratitis. Mice were pretreated by intraperitoneal injection of anti-mouse CD3u03B5. We observed the corneal infection of mice under the slit lamp microscope and made a clinical score. The protein expression of CD3u03B5 and interleukin-10 (IL-10) was determined by Western blotting. RESULTS With the disease progresses, the degree of corneal opacity and edema augmented. In the intra-stromal injection models, CD3u03B5 protein expression began to increase significantly on the 2nd day. However, in the scraping epithelial method models, CD3u03B5 only began to increase on the 3rd day. After natamycin treatment, the degree of corneal inflammation in mice was significantly attenuated on the 3rd day. After wedelolactone treatment, the severity of keratitis worsened. And the amount of CD3u03B5 protein was also reduced, compared with the control group. By inhibiting LOX-1 and Dectin-1, there was no significant difference in CD3u03B5 production compared with the control group. After inhibiting CD3u03B5, corneal ulcer area and clinical score increased, and IL-10 expression was downregulated. CONCLUSION As a pan T cell marker, CD3u03B5 participate in the adaptive immunity of A. fumigatus keratitis in mice. In our mice models, the corneas will enter the adaptive immune stage faster. By regulating IL-10, CD3u03B5 exerts anti-inflammatory and repairs effects in the adaptive immune stage.

AIM: To explore whether CD3ε is involved in the adaptive immunity of Aspergillus fumigatus (A. fumigatus) keratitis in mice and the role of innate and adaptive immunity in it. METHODS: Mice models of A. fumigatus keratitis were established by intra-stromal injection and corneal epithelial scratching. Subconjunctival injections of natamycin, wedelolactone, LOX-1 inhibitor (poly I) or Dectin-1 inhibitor (laminarin) were used to treat mice with A. fumigatus keratitis. Mice were pretreated by intraperitoneal injection of anti-mouse CD3ε. We observed the corneal infection of mice under the slit lamp microscope and made a clinical score. The protein expression of CD3ε and interleukin-10 (IL-10) was determined by Western blotting. RESULTS: With the disease progresses, the degree of corneal opacity and edema augmented. In the intra-stromal injection models, CD3ε protein expression began to increase significantly on the 2nd day. However, in the scraping epithelial method models, CD3ε only began to increase on the 3rd day. After natamycin treatment, the degree of corneal inflammation in mice was significantly attenuated on the 3rd day. After wedelolactone treatment, the severity of keratitis worsened. And the amount of CD3ε protein was also reduced, compared with the control group. By inhibiting LOX-1 and Dectin-1, there was no significant difference in CD3ε production compared with the control group. After inhibiting CD3ε, corneal ulcer area and clinical score increased, and IL-10 expression was downregulated. CONCLUSION: As a pan T cell marker, CD3ε participate in the adaptive immunity of A. fumigatus keratitis in mice. In our mice models, the corneas will enter the adaptive immune stage faster. By regulating IL-10, CD3ε exerts anti-inflammatory and repairs effects in the adaptive immune stage.

Background Diabetic retinopathy (DR) is the most common cause of diabetes-induced microvascular complications and it is the leading cause of blindness in working age worldwide. At the present stage,The main treatment for neovascularization and leakage in DR is anti-VEGF therapy, however, anti-VEGF therapy has its limitation,such as single target and short half-life of anti-VEGF drugs. Therefore, clarifying more therapeutic targets according to the molecular mechanism of neovascularization and leakage is needed, treating the disease by a drug which is multi-target and long-acting. Previous studies have shown that artesunate (ART) can inhibit retinal neovascularization and leakage through multiple targets. This study aimed to clarify the new mechanism of ART inhibiting retinal neovascularization and leakage. Objective To investigate the new mechanism of retinal neovascularization and leakage inhibited by artesunate (ART) . Methods Human Umbilical Vein Endothelial Cells (HUVEC) were divided into glucose (G) group, 40mmol/L G+ART(G40+ART)group, mannitol (M) control group, dimethyl sulfoxide (DMSO)control group. The concentration gradient of G group is 5.5mmol/L G (G5.5), 25mmol/L G (G25), 40mmol/L G (G40); The concentration gradient of M control group is 5.5 mmol / L G + 19.5 mmol / L M (M25), 5.5 mmol / L G + 34.5 mmol / L M (M40), The concentration gradient of ART of G40 + ART group is G40 + 10ug /ml ART(10A), G40+20ug/ml ART(20A), G40+40ug/ml ART(40A); the volume of DMSO in the DMSO control group is the same as it is in the 40A group. Western blot,and cell Immunofluorescence technique were used to detect the protein expression of ICAM-1 and MMP-9 in each group. Results Western blot,and cell Immunofluorescence showed that the protein expression of Intercellular adhesion molecule-1（ICAM-1）and Matrix metalloproteinase-9（MMP-9）in G25 group was higher than that in G5.5 group (P<0.01), and it increased in G40 group compared with G25 group (P<0.01);The protein expression of ICAM-1 and MMP-9 in G25 group was higher than that of M25 group (P<0.01),and it increased in G40 group compared with M40 (P<0.01);the protein expression of ICAM-1 and MMP-9 of G40+ART group was lower than that of G40 group,in which,it was lower in 20A group than that of 10A group (P<0.01), and it was lower in 40A group compared with 20A group (P<0.01). The DMSO control group showed that the protein expression of ICAM-1 and MMP-9 in G40+ART was lower than that of G40+DMSO group (P<0.01). Conclusion The two targets of ICAM-1 and MMP-9 may act as new therapeutic targets of ART to suppress the retinal neovascularization and leakage in DR,offering assistance for ART used in DR to treat the neovascularization and leakage.

Glaucoma, an insidious ophthalmic pathology, is typified by an aberrant surge in intraocular pressure (IOP) which culminates in the degeneration of retinal ganglion cells and optical neuropathy. The mitigation of IOP stands as the principal therapeutic strategy to forestall vision loss. The trabecular meshwork's (TM) integrity and functionality are pivotal in modulating aqueous humor egress. Despite their potential significance in glaucomatous pathophysiology, the implications of microRNAs (miRNAs) on TM functionality remain largely enigmatic. Transcriptomic sequencing was employed to delineate the miRNA expression paradigm within the limbal region of rodent glaucoma models, aiming to elucidate miRNA-mediated mechanisms within the glaucomatous milieu. Analytical scrutiny of the sequencing data disclosed 174 miRNAs with altered expression profiles, partitioned into 86 miRNAs with augmented expression and 88 with diminished expression. Notably, miRNAs such as hsa-miR-196b-5p were identified as having substantial expression discrepancies with concomitant statistical robustness, suggesting a potential contributory role in glaucomatous progression. Subsequent in vitro assays affirmed that miR-196b-5p augments the inflammatory cascade within immortalized human TM (iHTM) and glaucoma-induced human TM (GTM3) cells, concurrently attenuating cellular proliferation, motility, and cytoskeletal architecture. Additionally, miR-196b-5p implicates itself in the regulation of IOP and inflammatory processes in rodent models. At a mechanistic level, miR-196b-5p modulates its effects via the targeted repression of Nras (neuroblastoma RAS viral oncogene homolog). Collectively, these transcriptomic investigations furnish a comprehensive vista into the regulatory roles of miRNAs within the glaucomatous framework, and the identification of differentially expressed miRNAs alongside their targets could potentially illuminate novel molecular pathways implicated in glaucoma, thereby aiding in the development of innovative therapeutic avenues.

PurposesThe purpose of this meta-analysis is to systematically compare the efficacy and safety between combined surgery (phacoemulsification combined with trabeculectomy) and phacoemulsification for primary angle-closure glaucoma (PACG) patients.MethodsThis work was done through the data searched from the PubMed, EMBASE and the Cochrane Library. The Cochrane Handbook was also used to evaluate the quality of the included studies. In addition, this meta-analysis was performed using Revman 5.4 software.ResultsA total of seven randomized controlled trials (RCTs) were included. Compared with phacoemulsification alone, combined surgery resulted in significant reduction in the IOP (MD = -1.45, 95%CI, -2.38 to -0.51, p = .002). And more reduction in number of glaucoma medications was shown in the combined surgery group (MD, -0.59; 95%CI, -1.01 to -0.17; p = .006). For the improvement of the best corrected visual acuity (BCVA), there was no statistical differences between two groups (MD, 0.05; 95%CI, -0.01 to 0.11; p = .08). In subgroup analysis, the BCVA improved more significantly for medically controlled PACG patients in the phacoemulsification alone group (MD, 0.09; 95%CI, 0.01 to 0.07; p = .02). Compared with the phacoemulsification alone group, more complications were shown in the combined surgery group (RR, 0.22; 95%CI, 0.09 to 0.56; p = .001).ConclusionFor lowering IOP, combined surgery was superior to phacoemulsification alone for PACG patients. Meanwhile, combined surgery generated more complications. For medically controlled PACG patients, phacoemulsification alone is more conducive to the improvement of BCVA.

下肢慢性静脉功能不全(chronic venous insufficiency, CVI)的发病率越来越高,严重影响患者的生活质量.目前,CVI的主要病理生理机制为静脉反流、静脉回流障碍或者两者同时存在[1-2].CVI的治疗方式仍以压力治疗为主,但对于临床表现-病因学-解剖学-病理生理学(clinical etiology anatomy pathophysiology,CEAP)分级为C3级以上的病变[3],特别是存在髂股静脉闭塞的病变,腔内血管成形术和支架植入术已成为公认的首选治疗方法[4-5].目前,国内获批的静脉专用支架有Zilver Vena支架和Venovo支架,而青岛大学附属医院暂时无此两款支架,因此,对于髂股静脉全程闭塞性病变,基于静脉的特殊病理生理学特点及解剖位置,青岛大学附属医院尝试组合应用另外两种支架进行治疗.本研究对复合支架在髂股静脉闭塞性病变中的应用效果进行探讨,现报道如下.

Trabecular meshwork (TM) cell dysfunction is the leading cause of elevated intraocular pressure (IOP) and glaucoma. The long non-coding RNA (lncRNA) small nucleolar RNA host gene 11 (SNHG11) is associated with cell proliferation and apoptosis, but its biological functions and role in glaucoma pathogenesis remain unclear. In the present study, we investigated the role of SNHG11 in TM cells using immortalized human TM and glaucomatous human TM (GTM3 ) cells and an acute ocular hypertension mouse model. SNHG11 expression was depleted using siRNA targeting SNHG11. Transwell assays, quantitative real-time PCR analysis (qRT-PCR), western blotting, and CCK-8 assay were used to evaluate cell migration, apoptosis, autophagy, and proliferation. Wnt/β-catenin pathway activity was inferred from qRT-PCR, western blotting, immunofluorescence, and luciferase reporter and TOPFlash reporter assays. The expression of Rho kinases (ROCKs) was detected using qRT-PCR and western blotting. SNHG11 was downregulated in GTM3 cells and mice with acute ocular hypertension. In TM cells, SNHG11 knockdown inhibited cell proliferation and migration, activated autophagy, and apoptosis, repressing the Wnt/β-catenin signaling pathway, and activated Rho/ROCK. Wnt/β-catenin signaling pathway activity increased in TM cells treated with ROCK inhibitor. SNHG11 regulated Wnt/β-catenin signaling through Rho/ROCK by increasing GSK-3β expression and β-catenin phosphorylation at Ser33/37/Thr41 while decreasing β-catenin phosphorylation at Ser675. We demonstrate that the lncRNA SNHG11 regulates Wnt/β-catenin signaling through Rho/ROCK via β-catenin phosphorylation at Ser675 or GSK-3β-mediated phosphorylation at Ser33/37/Thr41, affecting cell proliferation, migration, apoptosis, and autophagy. Through its effects on Wnt/β-catenin signaling, SNHG11 is implicated in glaucoma pathogenesis and is a potential therapeutic target.

Glaucoma is a progressive, irreversible loss of retinal ganglion cells (RGCs) and axons that results in characteristic optic atrophy and corresponding progressive visual field defect. The exact mechanisms underlying glaucomatous neuron loss are not clear. The main risk factor for glaucoma onset and development is high intraocular pressure (IOP), however traditional IOP-lowering therapies are often not sufficient to prevent degeneration of RGCs and the vision loss may progress, indicating the need for complementary neuroprotective therapy. This review summarizes the progress for neuro protection in glaucoma in recent 5 years, including modulation of neuroinflammation, gene and cell therapy, dietary supplementation, and sustained-release system.

目的 探讨糖尿病视网膜病变(DR)、纤维蛋白原(FIB)及血细胞比容(HCT)与2型糖尿病(T2DM)患者发生颈动脉狭窄的关系.方法 收集2016年8月至2020年12月于青岛大学附属医院完成颈动脉超声检查并行眼底检查的175例T2DM患者的临床资料,根据颈动脉超声检查结果将其分为颈动脉狭窄组(n=86)和无狭窄组(n=89),分析T2DM患者发生颈动脉狭窄的影响因素.结果 单因素分析结果显示,高血压病史、DR、FIB、HCT均为T2DM患者发生颈动脉狭窄的影响因素(P<0.05).进一步将单因素分析中P<0.1的影响因素,包括年龄、高血压病史、DR、高密度脂蛋白胆固醇(HDL-C)、FIB及HCT纳入Logistic回归模型中进行多因素分析,结果显示,DR、HDL-C、FIB均是T2DM患者发生颈动脉狭窄的影响因素(P<0.05).狭窄组轻度、中度、重度狭窄或闭塞患者的FIB水平比较,差异均无统计学意义(P>0.05).Kendall's tau-b相关系数分析显示,DR分期与狭窄程度不存在相关性(r=-0.023,P=0.816).结论 DR、FIB、HCT、HDL-C、高血压病史均是T2DM患者发生颈动脉狭窄的影响因素,其中DR、FIB是T2DM患者发生颈动脉狭窄的独立影响因素.

新型降眼压药物Rho激酶抑制剂直接作用于青光眼根本的致病部位——小梁网及小梁网细胞,通过调节细胞周期、细胞骨架,促进细胞增殖等多种机制降眼压.本文将针对Rho激酶信号通路对小梁网细胞增殖作用机制的研究进展进行综述.

Aims: Elevated intraocular pressure is primarily induced by the increased resistance of conventional outflow of aqueous humor. Dysfunction of the juxtacanalicular region of trabecular meshwork (TM) and Schlemm's canal (SC) endothelium, as the main conventional outflow tissue, have been implicated as the major reasons for the increased resistance. Integrins are widespread in these tissues, especially alpha8 integrin (ITGA8). We aim to investigate the properties of cells expressing ITGA8 in the conventional outflow tissue. Main methods: Fluorescence in situ hybridization (FISH) and immunofluorescence (IF) were performed to detect the mRNA and protein levels of ITGA8 in human conventional outflow tissue. ITGA8-positive cells were isolated from the cultured human TM cells through a magnetic bead-based approach. Flow Cytometry was used to determine the purification efficiency. The expressions of TM and SC biomarkers and dexamethasone-induced myocilin secretion capacity of ITGA8-positive cells was assessed by Real-time PCR, IF and Western blot. A gel contraction assay was performed to evaluate contractility of ITGA8-positive cells after endothelin 1 treatment. Key findings: ITGA8 was found with robust expression near the inner wall of SC endothelium. After purification, the proportion of ITGA8-positive cells were increased by about 10%. ITGA8-positive cells were identified with the properties as SC endothelial cells, such as more robust expressions of SC biomarkers, less dexamethasoneinducible myocilin expression, and stronger contractility. Significance: This study demonstrated that cells expressing ITGA8 in SC region possess more properties as SC endothelial cells. Our data implicate a crucial role of ITGA8 in aqueous humor (AH) outflow resistance regulation.

Purpose Decreased trabecular meshwork (TM) cellularity has been implicated as a major reason for TM dysfunction and aqueous humor (AH) outflow abnormalities in primary open angle glaucoma. We previously found that transplantation of induced pluripotent stem cell (iPSC)-derived TM cells can restore TM function and stimulate endogenous TM cell division. The goal of the present study is to investigate whether signaling via gap junctions is involved in this process. Methods Differentiated iPSCs were characterized morphologically, transcriptionally, and immunohistochemically. After purification, iPSC-TM were co-cultured with mouse TM (MTM) cells to mimic the transplantation procedure. Through the pharmacological antagonists and short hairpin RNA (shRNA) technique, the gap junction function in iPSC-based therapy was determined. Results In the co-culture system, iPSC-TM increase MTM cell division as well as transfer of Ca2+ to MTM. This effect was blocked by treatment with the gap junction inhibitors carbenoxolone (CBX) or flufenamic acid (FFA). The shRNA mediated knock down of connexin 43 (Cx43) expression in iPSC-TM also results in decreased Ca2+ transfer and lower MTM proliferation rates. In vivo, Cx43 downregulation in transplanted iPSC-TM weakened their regenerative role in an Ad5.myocilinY437H mouse model of glaucoma. Mice receiving these cells exhibited lower TM cellularity and higher intraocular pressure (IOP) than those receiving unmodified iPSC-TM. Conclusions Our findings reveal a crucial role of gap junction, especially Cx43, in iPSC-based TM regeneration, and provides insights to enhance the regenerative effect of iPSCs in glaucoma therapy.

PURPOSE:Continuous reductions in trabecular meshwork (TM) cellularity inhibit aqueous humor (AH) outflow, which is the main cause of primary open-angle glaucoma. Rho-associated protein kinase inhibitor (ROCKi) targets the TM to reduce intraocular pressure (IOP) and increase AH outflow facility. However, the underlying mechanisms are not entirely clear. Here, we aimed to investigate the effect of a ROCKi (Y-27632) on TM cell proliferation and phagocytosis.METHODS:Immortalized human TM (iHTM) cells, glaucomatous TM (GTM3) cells, and primary human TM (pTM) cells were cultured and identified. The effects of various concentrations of Y-27632 on F-actin cytoskeleton were assessed using immunofluorescence. Cell proliferation effects were evaluated using a cell counting kit-8 (CCK8), cell counting, and Ki67 immunostaining. Cell phagocytosis was evaluated using immunofluorescence and flow cytometry in immortalized TM cells. C57BL/6J and Tg-MYOC Y437H mice were used to investigate the proliferative effects of Y-27632 on TM cells in vivo. The effect of Y-27632 on IOP was monitored for 2 weeks, and the outflow facility was detected 2 weeks after IOP measurement. TM cells in mice were counted using immunohistochemistry.RESULTS:Y-27632 (100 μM) significantly promoted the proliferation of both immortal TM cells and pTM cells. In GTM3 cells, phagocytosis was significantly greater in the Y-27632 group than in the control group, nearly reaching the level of phagocytosis in iHTM, as determined using immunofluorescence and flow cytometry. In Tg-MYOC Y437H mice, treatment with Y-27632 significantly decreased IOP and increased outflow facility, which greatly influenced the long-term IOP-lowering effect. The number of TM cells in Tg-MYOC Y437H mice was significantly improved after Y-27632 administration.CONCLUSION:Y-27632 promoted cell proliferation and phagocytosis of TM cells, and its proliferative effect was demonstrated in a transgenic mouse model. These results revealed a new IOP-lowering mechanism of Y-27632 through effects on TM cells, suggesting the potential for a correlation between TM cellularity and long-term recovery of IOP.

PurposeNerolidol, a naturally occurring sesquiterpene has both anti-microbial and anti-inflammatory properties. The current study aims to investigate the antifungal and the anti-inflammatory effects of nerolidol against mouse Aspergillus fumigatus (A. fumigatus) keratitis.MethodsThe minimum inhibitory concentration (MIC) and cytotoxicity tests were used to study the antifungal ability. For in vivo and in vitro studies, the mouse corneas and the human corneal epithelial cells (HCECs) infected with A. fumigatus spores were intervented with nerolidol or phosphate buffer saline (PBS). Thereafter, the effect of the nerolidol on the response against inflammation was analyzed using the following parameters: recruitment of the neutrophils or macrophages and the expression of the lectin-type oxidized low density lipoprotein receptor-1 (LOX-1) and interleukin 1β (IL-1β). Techniques used were the slit lamp, immunofluorescence, myeloperoxidase (MPO) detection, quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot.ResultsNerolidol directly inhibits the growth of A. fumigatus. The administration of nerolidol reduced the severity of fungal keratitis with infiltration of fewer inflammatory cells and reduced levels of the LOX-1, as well the anti-inflammatory cytokines such as IL-1β were reduced compared with the PBS group. Additionally, in vitro studies showed that treatment with nerolidol inhibited the production of the LOX-1 / IL-1β levels in A. fumigatus stimulated HCECs.ConclusionNerolidol attenuated the A. fumigatus keratitis inflammatory response by inhibiting the growth of A. fumigatus, reducing the recruitment of the neutrophils and the macrophages, and inhibiting the LOX-1/ IL-1β signaling.

Purpose Alginate oligosaccharides (AOS), obtained from depolymerizing alginate, has multiple pharmacological benefits. Cataract is a common disease caused by turbidity of the lens protein due to lens metabolism disorders. This study aimed to test the effects and the underlying mechanisms of AOS on D-galactose (D-gal)-mediated cataract. Materials and Methods A total of 45 8-week-old C57BL/6 J male mice were randomly divided into 5 groups. After eight weeks' intervention, the score of cataract was calculated depending on the turbidity of the lens. Hematoxylin and eosin (HE) and transmission electron microscope (TEM) images were observed. Superoxide dismutase (SOD) activity and malondialdehyde (MDA) level were measured by corresponding detection kits, respectively. SOD1, SOD2, catalase (CAT) and p53 protein expressions were examined by Western blot. Nuclear factor erythroid-2 related factor (Nrf2) and heme oxygenase-1 (HO-1) mRNA expressions were examined by Quantitative Real Time-PCR (RT-qPCR). Results The score of the turbidity of the lens showed that AOS significantly delayed the cataractogenesis. HE staining and TEM imaging showed that AOS decreased the damage and senescence of lenses in D-gal-induced C57BL/6 J mice. We further detected aging marker p53 expression in crystalline lenses, and our result showed that AOS significantly inhibited p53 protein expression in D-gal-induced mice. In addition, SOD activity and MDA level detection results showed that AOS significantly increased the activity of SOD, and decreased the level of MDA in crystalline lenses homogenates of D-gal-induced aging mice. Western blot results showed that AOS attenuated the damage of D-gal in the protein expressions of antioxidative enzymes SOD1, SOD2 and CAT. RT-qPCR results showed that AOS suppressed the down-regulation of Nrf2 and HO-1 mRNA expressions induced by D-gal. Conclusions AOS prevents against D-gal-mediated cataract in C57BL/6 J mice via inhibiting oxidative stress and up-regulating antioxidant system. Consequently, our results suggest that AOS may be an effective therapeutic strategy against cataract.

Glaucoma has been the leading cause of irreversible blindness worldwide. High intraocular pressure (IOP) is a high‐risk factor of glaucoma, repression of which has been the important treatment of glaucoma in clinic. Trabecular meshwork is crucial for maintaining IOP in aqueous humour out‐flow system. It is urgent to reveal the molecular mechanism of trabecular meshwork in glaucoma. Previous studies found that some pathways were related to glaucoma, such as extracellular matrix (ECM)‐receptor interaction, phosphatidylinositol 3‐kinase (PI3K)‐protein kinase B (Akt) and apoptosis. To identify novel molecules in glaucoma, we performed high‐throughput transcriptome and proteome analysis to immortal human trabecular meshwork cells (iHTM) and glaucomatous human trabecular meshwork cells (GTM3), respectively. Twenty‐six up‐regulated genes/proteins and 59 down‐regulated genes/proteins were identified as the high‐risk factors based on differential analysis, including some known factors of glaucoma. Furthermore, a glaucoma‐related protein‐protein interaction (PPI) network was constructed for investigating the function roles of risk factors. Some genes were identified as potential regulator in the pathogenesis of glaucoma based on the topology analysis and module analysis to the network. Importantly, we identified and demonstrated that CD9 played key roles in glaucoma by biological experiment. CD9 is down‐regulated in glaucoma, overexpression of CD9 can active integrin α4 (ITGA4), PI3K and Akt, which lead to the decreased apoptosis and attenuate glaucoma. All these results provide a novel molecular therapy of glaucoma.