Semaphorin 7A impairs barrier function in cultured human corneal epithelial cells in a manner dependent on nuclear factor-kappa B.

AIM: To evaluate the role of semaphorin 7A (Sema7A) and its associated regulatory mechanisms in modulating the barrier function of cultured human corneal epithelial cells (HCEs). METHODS: Barrier models of HCEs were treated with recombinant human Sema7A at concentrations of 0, 125, 250, or 500 ng/mL for 24, 48, or 72h in vitro. Transepithelial electrical resistance (TEER) as well as Dextran-fluorescein isothiocyanate (FITC) permeability assays were conducted to assess barrier function. To quantify tight junctions (TJs) such as occludin and zonula occludens-1 (ZO-1) at the mRNA level, reverse transcription-polymerase chain reaction (RT-PCR) analysis was performed. Immunoblotting was used to examine the activity of the nuclear factor-kappa B (NF-κB) signaling pathway and the production of TJs proteins. Immunofluorescence analyses were employed to localize the TJs. Enzyme-linked immunosorbent assay (ELISA) and RT-PCR were utilized to observe changes in interleukin (IL)-1ß levels. To investigate the role of NF-κB signaling activation and IL-1ß in Sema7A's anti-barrier mechanism, we employed 0.1 µmol/L IκB kinase 2 (IKK2) inhibitor IV or 500 ng/mL IL-1 receptor (IL-1R) antagonist. RESULTS: Treatment with Sema7A resulted in decreased TEER and increased permeability of Dextran-FITC in HCEs through down-regulating mRNA and protein levels of TJs in a time- and dose-dependent manner, as well as altering the localization of TJs. Furthermore, Sema7A stimulated the activation of inhibitor of kappa B alpha (IκBα) and expression of IL-1ß. The anti-barrier function of Sema7A was significantly suppressed by treatment with IKK2 inhibitor IV or IL-1R antagonists. CONCLUSION: Sema7A disrupts barrier function through its influence on NF-κB-mediated expression of TJ proteins, as well as the expression of IL-1ß. These findings suggest that Sema7A could be a potential therapeutic target for the diseases in corneal epithelium.

Snail Transcriptionally Represses Brachyury to Promote the Mesenchymal-Epithelial Transition in Ascidian Notochord Cells.

Mesenchymal-epithelial transition (MET) is a widely spread and evolutionarily conserved process across species during development. In Ciona embryogenesis, the notochord cells undergo the transition from the non-polarized mesenchymal state into the polarized endothelial-like state to initiate the lumen formation between adjacent cells. Based on previously screened MET-related transcription factors by ATAC-seq and Smart-Seq of notochord cells, Ciona robusta Snail (Ci-Snail) was selected for its high-level expression during this period. Our current knockout results demonstrated that Ci-Snail was required for notochord cell MET. Importantly, overexpression of the transcription factor Brachyury in notochord cells resulted in a similar phenotype with failure of lumen formation and MET. More interestingly, expression of Ci-Snail in the notochord cells at the late tailbud stage could partially rescue the MET defect caused by Brachyury-overexpression. These results indicated an inverse relationship between Ci-Snail and Brachyury during notochord cell MET, which was verified by RT-qPCR analysis. Moreover, the overexpression of Ci-Snail could significantly inhibit the transcription of Brachyury, and the CUT&Tag-qPCR analysis demonstrated that Ci-Snail is directly bound to the upstream region of Brachyury. In summary, we revealed that Ci-Snail promoted the notochord cell MET and was essential for lumen formation via transcriptionally repressing Brachyury.

Mixed-attitude three-way decision model for aerial targets: Threat assessment based on IF-VIKOR-GRA method.

Assessing potential threats typically necessitates the use of a robust mathematical model, a comprehensive evaluation method and universal decision rules. A novel approach is utilized in this study to optimize existing threat assessment (TA) algorithms and three-way decision models (3WDMs) are leveraged that incorporate decision-theoretic rough sets (DTRSs) within dynamic intuitionistic fuzzy (IF) environments to create a mixed-attitude 3WDM based on the IF-VIKOR-GRA method in the context of aviation warfare. The primary objectives of this study include determining conditional probabilities for IF three-way decisions (3WDs) and establishing mixed-attitude decision thresholds. Both the target attribute and loss function are expressed in the form of intuitionistic fuzzy numbers (IFNs). To calculate these conditional probabilities, an IF technique is used to combine the multi-attribute decision-making (MADM) method VIKOR and the grey relational analysis (GRA) method, while also taking into account the risk-related preferences of decision-makers (DMs). Optimistic and pessimistic 3WDMs are developed from the perspectives of membership degree and non-membership degree, then subsequently integrated into the comprehensive mixed-attitude 3WDM. The feasibility and effectiveness of this methodology are demonstrated through a numerical example and by comparison to other existing approaches.

17ß-estradiol inhibits TGF-ß-induced collagen gel contraction mediated by human Tenon fibroblasts via Smads and MAPK signaling pathways.

AIM: To investigate the impact of 17ß-estradiol on the collagen gels contraction (CGC) and inflammation induced by transforming growth factor (TGF)-ß in human Tenon fibroblasts (HTFs). METHODS: HTFs were three-dimensionally cultivated in type I collagen-generated gels with or without TGF-ß (5 ng/mL), 17ß-estradiol (12.5 to 100 µmol/L), or progesterone (12.5 to 100 µmol/L). Then, the collagen gel diameter was determined to assess the contraction, and the development of stress fibers was analyzed using immunofluorescence staining. Immunoblot and gelatin zymography assays were used to analyze matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) being released into culture supernatants. Enzyme-linked immunosorbent assay (ELISA) and reverse transcription-quantitative polymerase chain reaction (RT-PCR) were used to detect interleukin (IL)-6, monocyte chemoattractant proteins (MCP)-1, and vascular endothelial growth factor (VEGF) in HTFs at the translational and transcriptional levels. The phosphorylation levels of Sma- and Mad-related proteins (Smads), mitogen-activated protein kinases (MAPKs), and protein kinase B (AKT) were measured by immunoblotting. Statistical analysis was performed using either the Tukey-Kramer test or Student's unpaired t-test to compare the various treatments. RESULTS: The CGC caused by TGF-ß in HTFs was significantly inhibited by 17ß-estradiol (25 to 100 µmol/L), and a statistically significant difference was observed when comparing the normal control group with 17ß-estradiol concentrations exceeding 25 µmol/L (P<0.05). The suppressive impact of 17ß-estradiol became evident 24h after administration and peaked at 72h (P<0.05), whereas progesterone had no impact. Moreover, 17ß-estradiol attenuated the formation of stress fibers, and the production of MMP-3 and MMP-1 in HTFs stimulated by TGF-ß. The expression of MCP-1, IL-6, and VEGF mRNA and protein in HTFs were suppressed by 100 µmol/L 17ß-estradiol (P<0.01). Additionally, the phosphorylation of Smad2 Smad3, p38, and extracellular signal-regulated kinase (ERK) were downregulated (P <0.01). CONCLUSION: 17ß-estradiol significantly inhibits the CGC and inflammation caused by TGF-ß in HTFs. This inhibition is likely related to the suppression of stress fibers, inhibition of MMPs, and attenuation of Smads and MAPK (ERK and p38) signaling. 17ß-estradiol may have potential clinical benefits in preventing scar development and inflammation in the conjunctiva.

Tailor Traps in Bi3+-Doped NaGdGeO4 Phosphors by Introducing Eu3+ Ions to Switch Multimodal Phosphorescence Emission.

Featured with a tunable excitation/emission wavelength and excellent physicochemical stability, inorganic fluorescent materials are widely used in the fields of anti-counterfeiting. Here, we design a multi-stimuli-responsive dynamic fluorescence and phosphorescence anti-counterfeiting material by introducing Eu3+ ions in NaGdGeO4: Bi3+ to tailor the trap structure. The photoluminescence (PL), long persistent luminescence (LPL), and photo-stimulated luminescence (PSL) colors of NaGdGeO4: Bi3+, Eu3+ can be switched by varying the excitation modes (ultraviolet, near infrared, and X-ray light). Especially, the LPL and PSL colors of NaGdGeO4: Bi3+, Eu3+ vary with increasing decay and stimulation times. In addition, X-ray excitation ensures the specificity of the luminescence of NaGdGeO4: Bi3+, Eu3+ compared with ultraviolet excitation. This rapidly-changing-color fluorescent material offers the possibility of sophisticated anti-counterfeiting applications in the future.

Triterpenoid saponins from Psammosilene tunicoides and their antinociceptive activities.

Herein, five undescribed oleanane-type triterpenoid saponins, namely, psammosaponins A-E, along with nine known compounds, were isolated from the roots of Psammosilene tunicoides. Moreover, part of the ethanolic extract of P. tunicoides was acid-hydrolyzed and three aglycones were isolated from the resulting hydrolysate. The structures of all compounds were established through extensive analysis involving 1D and 2D NMR experiments, HRESIMS measurements, chemical derivatization, and comparison of spectroscopic data with the values reported in the literature. In all, 10 of the isolated saponins and the three aglycones were evaluated in the acetic acid-induced writhing model for their antinociceptive activity. At a dose of 40 mg/kg, these compounds exhibited significant inhibitory effects on the mouse writhing response, with inhibitions ranging from 31.9% to 79.3%. In addition, the structure-activity relationships of the isolates were discussed. Among the isolates, quillaic acid 3-O-glucuronide and 16α-hydroxygypsogenic acid showed better antinociceptive activity with inhibitions of 79.3% and 73.7%, respectively. Both isolates also exhibited antinociceptive activities in hot plate and formalin tests on mice. Their antinociceptive mechanism was explored in lipopolysaccharide-stimulated RAW 264.7 cells. These isolates could significantly inhibit the production of nitric oxide and interleukin-6 and downregulate the expression levels of inducible NO synthase, COX-1, and COX-2.

Luteolin ameliorates cornea stromal collagen degradation and inflammatory damage in rats with corneal alkali burn.

Corneal alkali burn (AB) is a blindness-causing ocular trauma commonly seen in clinics. An excessive inflammatory reaction and stromal collagen degradation contribute to corneal pathological damage. Luteolin (LUT) has been studied for its anti-inflammatory effects. In this study, the effect of LUT on cornea stromal collagen degradation and inflammatory damage in rats with corneal alkali burn was investigated. After corneal alkali burn, rats were randomly assigned to the AB group and AB + LUT group and received an injection of saline and LUT (200 mg/kg) once daily. Subsequently, corneal opacity, epithelial defects, inflammation and neovascularization (NV) were observed and recorded on Days 1, 2, 3, 7 and 14 post-injury. The concentration of LUT in ocular surface tissues and anterior chamber, as well as the levels of collagen degradation, inflammatory cytokines, matrix metalloproteinases (MMPs) and their activity in the cornea were detected. Human corneal fibroblasts (HCFs) were co-cultured with interleukin (IL)-1ß and LUT. Cell proliferation and apoptosis were assessed by CCK-8 assay and flow cytometry respectively. Measurement of hydroxyproline (HYP) in culture supernatants was used to quantify the amount of collagen degradation. Plasmin activity was also assessed. ELISA or real-time PCR was used to detect the production of matrix metalloproteinases (MMPs), IL-8, IL-6 and monocyte chemotactic protein (MCP)-1. Furthermore, the immunoblot method was used to assess the phosphorylation of mitogen-activated protein kinases (MAPKs), transforming growth factor-ß-activated kinase (TAK)-1, activator protein-1 (AP-1) and inhibitory protein IκB-α. At last, immunofluorescence staining helped to develop nuclear factor (NF)-κB. LUT was detectable in ocular tissues and anterior chamber after intraperitoneal injection. An intraperitoneal injection of LUT ameliorated alkali burn-elicited corneal opacity, corneal epithelial defects, collagen degradation, NV, and the infiltration of inflammatory cells. The mRNA expressions of IL-1ß, IL-6, MCP-1, vascular endothelial growth factor (VEGF)-A, and MMPs in corneal tissue were downregulated by LUT intervention. And its administration reduced the protein levels of IL-1ß, collagenases, and MMP activity. Furthermore, in vitro study showed that LUT inhibited IL-1ß-induced type I collagen degradation and the release of inflammatory cytokines and chemokines by corneal stromal fibroblasts. LUT also inhibited the IL-1ß-induced activation of TAK-1, mitogen-activated protein kinase (MAPK), c-Jun, and NF-κB signaling pathways in these cells. Our results demonstrate that LUT inhibited alkali burn-stimulated collagen breakdown and corneal inflammation, most likely by attenuating the IL-1ß signaling pathway. LUT may therefore prove to be of clinical value for treating corneal alkali burns.

The Conserved Transcriptional Activation Activity Identified in Dual-Specificity Tyrosine-(Y)-Phosphorylation-Regulated Kinase 1.

Dual-specificity tyrosine-(Y)-phosphorylation-regulated kinase 1 (DYRK1) encodes a conserved protein kinase that is indispensable to neuron development. However, whether DYRK1 possesses additional functions apart from kinase function remains poorly understood. In this study, we firstly demonstrated that the C-terminal of ascidian Ciona robusta DYRK1 (CrDYRK1) showed transcriptional activation activity independent of its kinase function. The transcriptional activation activity of CrDYRK1 could be autoinhibited by a repression domain in the N-terminal. More excitingly, both activation and repression domains were retained in HsDYRK1A in humans. The genes, activated by the activation domain of HsDYRK1A, are mainly involved in ion transport and neuroactive ligand-receptor interaction. We further found that numerous mutation sites relevant to the DYRK1A-related intellectual disability syndrome locate in the C-terminal of HsDYRK1A. Then, we identified several specific DNA motifs in the transcriptional regulation region of those activated genes. Taken together, we identified a conserved transcription activation domain in DYRK1 in urochordates and vertebrates. The activation is independent of the kinase activity of DYRK1 and can be repressed by its own N-terminal. Transcriptome and mutation data indicate that the transcriptional activation ability of HsDYRK1A is potentially involved in synaptic transmission and neuronal function related to the intellectual disability syndrome.

Magnesium deficiency stress in rice can be alleviated by partial nitrate nutrition supply.

The problem of nitrogen (N) excess and magnesium (Mg) deficiency in farmland is becoming more common, severe, and widespread in southern China. Magnesium is known to be an essential nutrient for higher plants; however, the physiological responses of field crops to Mg deficiency, particularly to its interaction with N forms, remain largely unknown. In this study, a hydroponic experiment was conducted using three Mg levels (0.01, 1.00, and 5.00 mM) and three nitrate/ammonium ratios (NO3-/NH4+ of 0/100, 25/75, and 50/50) under greenhouse conditions. The results show that Mg deficiency (0.01 mM) could result in yellow leaves, dwarf plants, and fewer tillers during rice growth. Furthermore, Mg deficiency induced a major reduction in root morphology and activity, photosynthetic properties, and nutrient accumulation, while it resulted in a clear increase in malondialdehyde, superoxide dismutase, peroxidase, and catalase activities in rice. However, under Mg-deficiency stress, the supply of partial NO3- led to a significant drop in these antioxidant enzyme activities. Moreover, partial NO3- supply significantly improved the net photosynthetic rate, transpiration rate, stomatal conductance, and intercellular CO2 concentrations under Mg-deficiency conditions. In particular, the supply of partial NO3- dramatically promoted the growth of the root system, boosted the occurrence of lateral roots, and enhanced root vitality under Mg-deficiency stress. Additionally, the supply of partial NO3- led to significant increases in dry weight and N and Mg contents under Mg deficiency. The results of this study suggest that the symptoms of Mg-deficiency stress in rice can be alleviated by partial NO3- supply.

Mechanoluminescence from an Ion-Irradiated Single Crystal of Lithium Niobium Oxide.

Mechanoluminescence (ML) is a well-known phenomenon that has a wide range of applications in security monitoring, biomechanical sensing, and displays. Although several mechanisms relating to ML have been proposed, significant ambiguity persists due to the coexistence of crystal boundaries, luminescence centers, and defects within the samples, making them hard to disentangle. Here we preclude such ambiguity by using a Kr+-irradiated single crystal of lithium niobium oxide (LiNbO3) as the ML materials so that oxygen vacancies are retained to modulate the ML properties. We explore the ion concentration- and species-dependent ML properties along with the band calculations to explicitly reveal that it is the trapped electrons at the oxygen vacancies that are transferred to the conduction band under the piezopotentials of LiNbO3, which combine with holes in the valence band and emit photons. This in-depth understanding not only clarifies the long-standing obscurity of the ML mechanism but also paves a rational and scalable way for the design of advanced ML materials with superior performances.

Inhibitory effects of luteolin on TLR3-mediated inflammation caused by TAK/NF-κB signaling in human corneal fibroblasts.

AIM: To study the role of luteolin (LUT) in the expression of toll-like receptors 3 (TLR3) ligand polyI:C stimulated inflammatory factors in human corneal fibroblasts (HCFs). METHODS: HCFs cells were cultivated with or without LUT or polyI:C. The expression levels of interleukin (IL)-6, IL-8, monocyte chemotactic protein-1 (MCP-1), vascular cell adhesion molecule (VCAM)-1, as well as intercellular adhesion molecule (ICAM)-1 were measured using enzyme-linked immunosorbent assay (ELISA), immunoblotting or reverse transcription-quantitative polymerase chain reaction (PCR) analyses. Immunoblotting was used to assess toll-interleukin-1 receptor-domain-containing adapter-inducing interferon-ß (TRIF), TLR3, transforming growth factor-b-activated kinase 1 (TAK1), tumor necrosis factor receptor-associated factor 6 (TRAF6), the transcription factor AP-1, as well as transcription factor nuclear factor (NF-κB)-inhibitory protein IκB-α degradation and phosphorylation. Immunofluorescence assays were used to localize the cellular location of the p65 subunit of NF-κB. RESULTS: Corneal fibroblasts exposed to polyI:C demonstrated decreased VCAM-1, ICAM-1, MCP-1, IL-6, and IL-8 expression levels upon exposure to LUT in a time-dependent and concentration-dependent manner. LUT was observed to suppress polyI:C-triggered expression of TLR3, the translocation of NF-κB p65 into cell nuclei, as well as the phosphorylation of TAK, c-Jun, and IκB-α, while no impact on the expression levels of TRIF and TRAF6 were observed. CONCLUSION: LUT suppress the expression of proinflammatory adhesion molecules, chemokines, and cytokines in polyI:C exposed HCFs. These effects are likely mediated through TAK/NF-κB signal attenuation. Therefore, LUT is a candidate molecule that can prevent the TLR3-mediated inflammation response associated with corneal viral infection.

Sulforaphane inhibits cytokine-stimulated chemokine and adhesion molecule expressions in human corneal fibroblasts: Involvement of the MAPK, STAT, and NF-κB signaling pathways.

Chemokines and adhesion molecules are major inflammatory mediators of chronic and recurrent vernal keratoconjunctivitis (VKC). Sulforaphane (SFN) is a natural plant extract that is known to have anti-inflammatory and antioxidant properties. SFN is demonstrated to be effective against a variety of human diseases. The current investigation examines the effects and the molecular mechanisms of SFN on cytokine-induced human corneal fibroblasts (HCFs) expression of adhesion molecules and chemokines. HCFs were exposed to both interleukin (IL)-4 and tumor necrosis factor (TNF)-α in the absence or presence of SFN treatment. The levels of thymus- and activation-regulated chemokine (TARC) and eotaxin-1 in culture supernatants were evaluated using enzyme-linked immunosorbent assay (ELISA). Reverse transcription-polymerase chain reaction analysis (RT-PCR) enabled quantification of mRNA levels of vascular cell adhesion molecule (VCAM)-1, eotaxin-1, and TARC along with cytokine receptors. An immunoblotting assay was used to evaluate the activities of VCAM-1, nuclear factor-kappa B (NF-κB), mitogen-activated protein kinases (MAPKs), signal transducer and activator of transcription factor (STAT)6 pathways, along with the expression of the cytokine receptors including IL-4 receptor (R)α, IL-13Rα1, TNFRI, as well as TNFRII. SFN inhibited TARC and eotaxin-1 release in HCFs stimulated by TNF-α and IL-4 in a manner dependent on dose and time. SFN suppressed transcriptions of TARC, eotaxin-1, and VCAM-1. Furthermore, the mRNA and protein expression levels of IL-4Rα, TNFRI, and TNFRII were also attenuated by SFN exposure, however, those of IL-13Rα1 remained unaffected. In addition, SFN downregulated the expression of VCAM-1 and the phosphorylation of MAPKs, IκBα, and STAT6. These results suggest that SFN inhibited cytokine-stimulated TARC, eotaxin-1 secretion as well as VCAM-1 expression in HCFs, with these effects likely occurring as a result of cytokine receptor inhibition and attenuation of MAPK, NF-κB, and STAT6 signaling. SFN may therefore have therapeutic potential in VKC treatment.

Effective Repeatable Mechanoluminescence in Heterostructured Li1- x Nax NbO3 : Pr3.

Mechanoluminescence (ML) is a striking optical phenomenon that is achieved through mechanical to optical energy conversion. Here, a series of Li1 -x Nax NbO3 : Pr3+ (x = 0, 0.2, 0.5, 0.8, 1.0) ML materials have been developed. In particular, due to the formation of heterostructure, the synthesized Li0.5 Na0.5 NbO3 : Pr3+ effectively couples the trap structures and piezoelectric property to realize the highly repeatable ML performance without traditional preirradiation process. Furthermore, the ML performances measured under sunlight irradiation and preheating confirm that the ML properties of Li0.5 Na0.5 NbO3 : Pr3+ can be ascribed to the dual modes of luminescence mechanism, including both trap-controllable and self-recoverable modes. In addition, DFT calculations further confirm that the doping of Na+ ions in LiNbO3 leads to electronic modulations by the formation of the heterostructures, which optimizes the trap distributions and concentrations. These modulations improve the electron transfer efficiency to promote ML performances. This work has supplied significant references for future design and synthesis of efficient ML materials for broad applications.

Inhibition of TGF-ß2-induced migration and epithelial-mesenchymal transition in ARPE-19 by sulforaphane.

AIM: To investigate the effects of sulforaphane (SFN) on transforming growth factor (TGF)-ß2 stimulated migration and epithelial-mesenchymal transition (EMT) in ARPE-19 cells. METHODS: ARPE-19 cells were cultured in the presence or absence of SFN or TGF-ß2. SFN toxicity was assessed by performing a lactate dehydrogenase assay (LDH) and 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assays, and cell migration was evaluated by Transwell migration assay. Actin stress fiber formation in ARPE-19 cells was determined using immunofluorescence analysis. Immunoblotting analysis was used to determine fibronectin and α-smooth muscle actin expressions along with the degree of Smad and Akt phosphorylation. RESULTS: SFN inhibited ARPE-19 migration. Additionally, SFN attenuated TGF-ß2-induced appearance of actin stress fibers as well as fibronectin and α-smooth muscle actin expressions in these cells. SFN also hindered the TGF-ß2-stimulated phosphorylation of Smad2, Smad3, and Akt. SFN showed no cytotoxicity towards ARPE-19 cells. CONCLUSION: SFN inhibits TGF-ß2-stimulated migration and EMT in ARPE-19 cells, probably by preventing the establishment of actin stress fibers and Akt and Smad2/3 signaling.

Sulforaphane suppresses polyinosinic­polycytidylic acid­stimulated release of cytokines, chemokines and MMPs by human corneal fibroblasts.

Viral corneal infection is a common cause of visual impairment and blindness. Polyinosinic­polycytidylic acid, or poly(I:C), is similar to viral double­stranded RNA in structure and has been implicated in the release of a variety of cytokines, chemokines and matrix metalloproteinases (MMPs) by corneal fibroblasts. Sulforaphane (SFN) is an isothiocyanate compound found in cruciferous vegetables. The present study investigated the potential effect of SFN on the poly(I:C)­stimulated release of cytokines, chemokines and MMPs in human corneal fibroblasts (HCFs). ELISA showed that SFN was associated with a time­ and dose­dependent reduction in poly(I:C)­stimulated production of interleukin (IL)­8, chemoattractant protein­1, IL­6, MMP­1 and MMP­3 by HCFs. Western blot analysis indicated that SFN suppressed the function of poly(I:C) by modulating mitogen­activated protein kinases (MAPKs), including p38 and extracellular signal­regulated kinase (ERK), activator protein­1 (AP­1) component c­Jun and the kinase, Akt, and the phosphorylation and degradation of the nuclear factor (NF)­κB inhibitor IκB­α. Immunofluorescence analysis revealed that SFN attenuated the production of poly(I:C)­induced nuclear translocation of the NF­κB p65 subunit. Reverse transcription­quantitative PCR analysis revealed that SFN prevented the poly(I:C)­induced upregulation of Toll­like receptor 3 (TLR3) mRNA expression in HCFs. No significant cytotoxic effect of SFN on HCFs was observed. In summary, SFN attenuated the poly(I:C)­induced production of proinflammatory chemokines, cytokines and MMPs by HCFs, by inhibiting TLR3, MAPK (p38 and ERK), AP­1, Akt and NF­κB signaling. SFN may therefore be a potential novel treatment for viral corneal infection by limiting immune cell infiltration.

Alarmins from conjunctival fibroblasts up-regulate matrix metalloproteinases in corneal fibroblasts.

AIM: To explore the effects of alarmins produced by necrotic human conjunctival fibroblasts on the release of matrix metalloproteinases (MMPs) by human corneal fibroblasts (HCFs). METHODS: A necrotic cell supernatant (NHCS) was prepared by subjecting human conjunctival fibroblasts to three cycles of freezing and thawing. The amounts of interleukin (IL)-1ß and tumor necrosis factor (TNF)-α in NHCS were determined by enzyme-linked immunosorbent assays. HCFs exposed to NHCS or other agents in culture were assayed for the release of MMPs as well as for intracellular signaling by immunoblot analysis. The abundance of MMP mRNAs in HCFs was examined by reverse transcription and real-time polymerase chain reaction analysis. RESULTS: NHCS increased the release of MMP-1 and MMP-3 by HCFs as well as the amounts of the corresponding mRNAs in the cells. NHCS also induced activation of mitogen-activated protein kinase (MAPK) signaling pathways mediated by extracellular signal-regulated kinase (ERK), p38, and c-Jun NH2-terminal kinase (JNK) as well as elicited that of the nuclear factor (NF)-κB signaling pathway by promoting phosphorylation of the endogenous NF-κB inhibitor IκB-α. Inhibitors of MAPK and NF-κB signaling as well as IL-1 and TNF-α receptor antagonists attenuated the NHCS-induced release of MMP-1 and MMP-3 by HCFs. Furthermore, IL-1ß and TNF-α were both detected in NHCS, and treatment of HCFs with these cytokines induced the release of MMP-1 and MMP-3 in a concentration-dependent manner. CONCLUSION: Alarmins, including IL-1ß and TNF-α, produced by necrotic human conjunctival fibroblasts triggered MMP release in HCFs through activation of MAPK and NF-κB signaling. IL-1ß and TNF-α are therefore potential therapeutic targets for the amelioration of corneal stromal degradation in severe ocular burns.

Achieving high thermal stability of different rare-earth ions in a single matrix host via the manipulation of the local structure by a solid solution.

Thermal quenching seriously restricts the practical application of phosphors, particularly under high temperature and long-term working conditions. Here, we demonstrate that the as-obtained series of solid solutions of Ca2-xYxAl2Si1-xAlxO7:Tb3+ (x = 0-1, Ca2Al2SiO7 → CaYAl3O7) phosphors exhibit an adjustable optical performance, where CaYAl3O7:Tb3+ exhibits a greatly improved thermal stability with a shortened bond distance of the related polyhedron compared with Ca2Al2SiO7:Tb3+. The shrunken bond distance strengthens the pressure of the local structure and suppresses the non-radiative transition effectively, contributing to the prevention of the thermal degradation. The formed phosphor with excellent structural stability could be effectively incorporated with various lanthanide ions (Eu3+, Tb3+, Sm3+, Dy3+, and Pr3+) to address a pleochroism output.

Genomic basis of environmental adaptation in the leathery sea squirt (Styela clava).

Tunicates occupy the evolutionary position at the boundary of invertebrates and vertebrates. It exhibits adaptation to broad environmental conditions and is distributed globally. Despite hundreds of years of embryogenesis studies, the genetic basis of the invasive habits of ascidians remains largely unknown. The leathery sea squirt, Styela clava, is an important invasive species. We used the chromosomal-level genome and transcriptome of S. clava to explore its genomic- and molecular-network-based mechanisms of adaptation to environments. Compared with Ciona intestinalis type A (C. robusta), the size of the S. clava genome was expanded by 2-fold, although the gene number was comparable. An increase in transposon number and variation in dominant types were identified as potential expansion mechanisms. In the S. clava genome, the number of genes encoding the heat-shock protein 70 family and members of the complement system was expanded significantly, and cold-shock protein genes were transferred horizontally into the S. clava genome from bacteria. The expanded gene families potentially play roles in the adaptation of S. clava to its environments. The loss of key genes in the galactan synthesis pathway might explain the distinct tunic structure and hardness compared with the ascidian Ciona species. We demonstrated further that the integrated thyroid hormone pathway participated in the regulation of larval metamorphosis that provides S. clava with two opportunities for adapting to their environment. Thus, our report of the chromosomal-level leathery sea squirt genome provides a comprehensive genomic basis for the understanding of environmental adaptation in tunicates.

Inhibition of Gap Junction-Mediated Intercellular Communication by Poly(I:C) in Cultured Human Corneal Fibroblasts.

PURPOSE/AIM: Corneal stromal fibroblasts are connected to each other via gap junctions, which contribute to maintenance of corneal homeostasis. Viral infection of the corneal stroma can result in inflammation and scarring. The effects of polyinosinic-polycytidylic acid [poly(I:C)], an analog of viral double-stranded RNA, on gap junctional intercellular communication (GJIC) in cultured human corneal fibroblasts (HCFs) were examined. MATERIALS AND METHODS: Cultured HCFs were exposed to poly(I:C) in the absence or presence of inhibitors of mitogen-activated protein kinase (MAPK) signaling or the antioxidant N-acetyl-L-cysteine (NAC). Expression of the gap junction protein connexin 43 (Cx43) was examined by immunoblot and immunofluorescence analyses. The level of Cx43 mRNA or microRNA-21 or -130a was determined by quantitative reverse transcription-polymerase chain reaction analysis. GJIC was measured with a dye coupling assay. The amount of malondialdehyde and the activity of superoxide dismutase (SOD) were measured with assay kits. RESULTS: Exposure of HCFs to poly(I:C) resulted in down-regulation of Cx43 expression and GJIC activity as well as in up-regulation of microRNA-21 expression. Poly(I:C) increased the amount of malondialdehyde and reduced the activity of SOD in the cells, and these effects were prevented by NAC. The inhibitory effects of poly(I:C) on both Cx43 expression and GJIC activity were attenuated by NAC and by c-Jun NH2-terminal kinase (JNK) inhibitor II. CONCLUSIONS: Poly(I:C) inhibited Cx43 expression and GJIC in cultured HCFs, possibly as a result of the associated up-regulation of microRNA-21. Poly(I:C) also increased oxidative stress in these cells, and such stress together with signaling by the MAPK JNK was implicated in the effects of poly(I:C) on Cx43 expression and GJIC activity. Down-regulation of GJIC activity among corneal fibroblasts by double-stranded RNA may thus contribute to the disruption of stromal homeostasis during viral infection of the cornea.

Detection of Cell Viability via Fluorescence Labeling of Silicate Phosphor with a Low-Temperature Superlong Persistent Luminescence.

Cell viability detection is usually accompanied by large errors and toxicity. For the accuracy and safety of detection, it is urgent to develop a practical approach to realize the real-time monitoring of cell viability. Here, a low-temperature persistent phosphor was employed to facilitate the detection of cell viability. Stable low-temperature persistent luminescence originating from Ba4Si6O16:Eu2+, Pr3+ (BSEP) was developed, which could be quantified to describe the viability of baby Syrian hamster kidney (BHK-21) cells. The defect state structure guarantees the excellent persistent luminescence at 193 K of more than 200 h of the as-prepared BSEP phosphor, which ensures the long-term biological application. The structural thermal stability and water tolerance of BSEP suggest the feasibility of the practical application. Our results provide the underlying insights of the low-temperature persistent phosphors for real-time biological detection.