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

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International Journal of Ophthalmology(English edition) 2022年第3期15卷 371-379页

作者： Zi-Han Guo ping-ping liu Heng Wang Xiu-Xia Yang Cheng-Cheng Yang Hui Zheng Dan Tang Yang liu Department of Ophthalmology the Fifth Affiliated HospitalSun Yat-sen UniversityZhuhai 519000Guangdong ProvinceChina

AIM: To study the role of luteolin(LUT) in the expression of toll-like receptors 3(TLR3) ligand poly I:C stimulated inflammatory factors in human corneal fibroblasts(HCFs).METHODS: HCFs cells were cultivated with or without LUT or poly I: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 enzymelinked 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-β(TRIF), TLR3, transforming growth factor-bactivated 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 poly I: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 poly I: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 poly I: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.

关键词： toll-like receptor inflammation corneal fibroblast poly I:C

Microstructure,hardening and deuterium retention in CVD tungsten irradiated with neutrons at temperatures of defect recovery stagesⅡandⅢ

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Tungsten 2022年第3期4卷 248-260页

作者： Xiao-Ou Yi Tatsuya Kuwabara Vladimir KhAlimov Yu-Feng Du Wen-Tuo Han ping-ping liu Bin-You Yan Jiu-Peng Song Kenta Yoshida Takeshi Toyama Fa-Rong Wan Somei Ohnuki Yuji Hatano Yasuyoshi Nagai School of Materials Science and Engineering University of Science and Technology BeijingBeijing100083China Graduate School of Engineering Nagoya UniversityFuro-choChikusa-kuNagoya464-8063Japan Frumkin Institute of Physical Chemistry and Electrochemistry Russian Academy of SciencesMoscow119071Russia Institute for Materials Research Tohoku UniversityNarita-cho 2145-2OaraiIbaraki311-1313Japan Xiamen Tungsten Co.Ltd. Xiamen361021China School of Materials Science and Engineering Xihua UniversityChengdu610039China Hydrogen Isotope Research Center University of ToyamaToyama930-8555Japan

Samples of ultra-high-purity tungsten prepared using chemical vapour deposition(CVD)technique were irradiated with neutrons at temperatures T_(irr)=373-483 K(stage Ⅱ of defect recovery)and T_(irr)=573-673 K(stage Ⅲ)up to 0.15 displacements per atom(dpa)in the Belgian reactor(BR2).The study of the microstructure of neutron-damaged samples using transmission electron microscopy(TEM)revealed visible defects with a predominance of dislocation loops.With an increase in the neutron irradiation temperature,the spatial distribution of the loops acquired pronounced inhomogeneity,and their average size moderately increased.Cavities and voids were not observed.Irradiation-induced hardening was found and a linear correlation was obtained between Vickers microhardness and nanohardness for undamaged and neutron-irradiated CVD-W samples.Irradiation of tungsten with neutrons led to a significant increase in the retention of deuterium,which accumulated mainly in vacancy-type traps.Furthermore,the influence of the columnar grain structure in low-dose neutron-irradiated tungsten seemed to be non-trivial upon deuterium retention.

关键词： CVD-W Neutron irradiation Microstructure Hardening Deuterium retention

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

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International Journal of Ophthalmology(English edition) 2020年第7期13卷 1031-1038页

作者： Lin Chen Ye liu Xiao-Shuo Zheng Hui Zheng ping-ping liu Xiu-Xia Yang Yang liu Department of Ophthalmology the Fifth Affiliated HospitalSun Yat-sen UniversityZhuhai519000Guangdong ProvinceChina Department of Pathology the Fifth Affiliated HospitalSun Yat-sen UniversityZhuhai 519000Guangdong ProvinceChina

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)-1βand tumor necrosis factor(TNF)-αin 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 m RNAs 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 m RNAs 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-αreceptor antagonists attenuated the NHCS-induced release of MMP-1 and MMP-3 by HCFs.Furthermore,IL-1βand TNF-αwere 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-1βand TNF-α,produced by necrotic human conjunctival fibroblasts triggered MMP release in HCFs through activation of MAPK and NF-κB signaling.IL-1βand TNF-αare therefore potential therapeutic targets for the amelioration of corneal stromal degradation in severe ocular burns.

关键词： alarmin ocular burn matrix metalloproteinase corneal fibroblast conjunctival fibroblast inflammation

Inhibition of TGF-β2-induced migration and epithelial-mesenchymal transition in ARPE-19 by sulforaphane

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International Journal of Ophthalmology(English edition) 2021年第7期14卷 973-980页

作者： Yan-Bing Huang ping-ping liu Hui Zheng Xiu-Xia Yang Cheng-Cheng Yang Ye liu Yang liu Department of Ophthalmology the Fifth Affiliated HospitalSun Yat-sen UniversityZhuhai 519000Guangdong ProvinceChina Department of Pathology the Fifth Affiliated HospitalSun Yat-sen UniversityZhuhai 519000Guangdong ProvinceChina

AIM: To investigate the effects of sulforaphane(SFN) on transforming growth factor(TGF)-β2 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-β2. SFN toxicity was assessed by performing a lactate dehydrogenase assay(LDH) and 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2 H-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-β2-induced appearance of actin stress fibers as well as fibronectin and α-smooth muscle actin expressions in these cells. SFN also hindered the TGF-β2-stimulated phosphorylation of Smad2, Smad3, and Akt. SFN showed no cytotoxicity towards ARPE-19 cells. CONCLUSION: SFN inhibits TGF-β2-stimulated migration and EMT in ARPE-19 cells, probably by preventing the establishment of actin stress fibers and Akt and Smad2/3 signaling.

关键词： transforming growth factor-β2 epithelial-mesenchymal transition sulforaphane ARPE-19

Dynamic changes in the radiologic manifestation of a recurrent checkpoint inhibitor related pneumonitis in a non-small cell lung cancer patient:A case report

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World Journal of Clinical Cases 2021年第30期9卷 9108-9113页

作者： Pei-Xin Tan Wei Huang ping-ping liu Yi Pan Yan-Hai Cui Department of Radiation Oncology Guangdong Provincial People's Hospital and Guangdong Academy of Medical SciencesGuangzhou 510080Guangdong ProvinceChina

BACKGROUND As immune checkpoint inhibitors(ICIs)have become widely used in lung cancer treatment,immune-related adverse events(irAEs)warrant sufficient attention.Checkpoint inhibitor-related pneumonitis(CIP)is one of the most concerning adverse events as it is uncommon but life threatening.CASE SUMMARY The patient whose case is reported here experienced three episodes of CIP in a span of 4 mon.Interestingly,the three episodes of CIP involved different regions of the lung separately.Taking these pneumonitis areas together makes nearly a whole lung area.CONCLUSION This case showed that recurrent CIPs may occur repeatedly until the whole lung is involved,suggesting that the follow-up period of CIP should be long enough,and the rechallenge of ICI should be done with due caution.

关键词： Checkpoint inhibitor-related pneumonitis Lung cancer Radiology Immune checkpoint inhibitors Recurrent pneumonitis Case report

Triptolide inhibits TGF-β-induced matrix contraction and fibronectin production mediated by human Tenon fibroblasts

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International Journal of Ophthalmology(English edition) 2018年第7期11卷 1108-1113页

作者： Yang liu ping-ping liu Lei liu Xiao-Shuo Zheng Hui Zheng Cheng-Cheng Yang Ci-Ren Luobu Ye liu Department of Ophthalmology the Fifth Affiliated Hospital Sun Yat-sen University Department of Ophthalmology the First Hospital of Jilin University Department of Pathology the Fifth Affiliated Hospital Sun Yat-sen University

AIM： To determine if triptolide influences the contractility and fibronectin production in human Tenon fibroblasts（HTFs）. METHODS： HTFs were cultured in type I collagen gels with or without transforming growth factor beta（TGF-β） and/or triptolide. The diameter of the collagen gel was used to measure contraction. Immunoblot analysis was used to quantify myosin light chain（MLC） phosphorylation and integrin expression. Laser confocal fluorescence microscopy was used to monitor the formation of actin stress fibers. Fibronectin production was measured with an enzyme immunoassay. RESULTS： Triptolide inhibition of contraction in TGF-β-induced collagen gel mediated by HTFs was dosedependent and statistically significant at 3 nmol/L（P〈0.05） and maximal at 30 nmol/L and significantly time dependent at 2 d（P〈0.05）. Triptolide reduced TGF-β-induced expression of integrins α5 and β1, phosphorylation of MLC, and formation of stress fibers in HTFs. Furthermore, the inhibition of triptolide on the attenuated TGF-β-induced production of fibronectin by HTFs was concentration-dependent and significant at 1 nmol/L（P〈0.05） and maximal at 30 nmol/L. CONCLUSION： Triptolide suppress the contractility of HTFs induced by TGF-β and the production of fibronectin by these cells. It is promising that triptolide treatment may possibly inhibit scar formation after glaucoma filtration surgery.

关键词： Tenon fibroblast triptolide transforming growth factor β wound healing fibronectin

Microstructure characterization in a sensitized Al-Mg-Mn-Zn alloy

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Rare Metals 2018年第2期37卷 129-135页

作者： Chun-Yan Meng Di Zhang ping-ping liu Lin-Zhong Zhuang Ji-Shan Zhang State Key Laboratory for Advanced Metals and Materials University of Science and Technology Beijing School of Materials Science and Engineering University of Science and Technology Beijing

Alloying elements, present in the aluminum solid solution or the precipitates, influence the corrosion resistance of A1-Mg-Mn-Zn alloys. In this study, sensi- tizing treatment was applied to an A1-Mg-Mn-Zn alloy to modify the precipitation at the grain boundaries or in the grains. Transmission electron microscopy （TEM） and scanning electron microscope （SEM） were used to characterize various second-phase particles and determine their orientation relationship with the A1 matrix. After sensitizing treatment, z-phase （Mg32（Al, Zn）49） is observed to precipitate along the grain boundaries in a coarser size, producing a discontinuous grain boundary precipitate structure. In addition, Mn-rich particles are found to form with various shapes, such as global, plate and rhombus.

关键词： Al-Mg-Mn-Zn alloy Mg32（A1, Zn）49 phase A16（Mn, Fe） phase Microstructure characterization

Microstructure and Nano-hardness of Pure Copper and ODS Copper Alloy under Au Ions Irradiation at Room Temperature

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Acta Metallurgica Sinica(English Letters) 2016年第11期29卷 1047-1052页

作者： Jing Zhang Yong-Qin Chang Zhi-Meng Guo ping-ping liu Yi Long Fa-Rong Wan School of Materials Science and Engineering University of Science and Technology Beijing

The microstructure and nano-hardness of the pure copper and oxide dispersion-strengthened（ODS） copper alloy subjected to 1.4 Me V Au ions irradiation at room temperature were investigated. After irradiation, dislocation-loops form in both materials, while voids can only be generated in the pure copper. Compared with the irradiated pure copper, larger average diameter and lower number density of irradiation-induced dislocation-loops were detected in the ODS copper alloy, revealing that high-density dislocation and large volume of Al2O3 particles existing in the ODS copper alloy can act as effective sinks for the irradiation-induced defects. It was also detected that irradiation hardening in the ODS copper alloy is lower than that in the pure copper. The microstructure and nano-hardness results reveal that the ODS copper alloy has a better irradiation tolerance than the pure copper. In addition, the average diameter of the Al2O3 particles in the ODS copper alloy decreases after irradiation, because the Al–O chemical bonds are decomposed and the atoms are redistributed in the matrix during the irradiation process. This work reveals that the irradiation tolerance of the copper can be effectively enhanced by adding nano-sized Al2O3 particles into the matrix.

关键词： Oxide dispersion-strengthened copper alloy Ions irradiation Microstructure evolution Nano-hardness

Effect of zymosan on the expression and function of the gap-junction protein connexin 43 in human corneal fibroblasts

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International Journal of Ophthalmology(English edition) 2021年第3期14卷 341-348页

作者： Xiao-Shuo Zheng Hui Zheng Dan Xu ping-ping liu Bing Li Zi-Mu Cao Yang liu Ye liu Department of Ophthalmology the Fith Affiliated HospitalSun Yat-sen UniversityZhuhai 5190001 Guangzhou ProvinceChina Institute of Enironmental Systems Biology Environmental Science and Enginecring CollegeDalian Maritime UniversityDalian 116027Liaoning ProvinceChina Department of Pathology the Fifth Affiliated HospitalSun Yat-sen UniversityZhuhai 519000Guangzhou ProvinceChina

AIM:To study the effect of zymosan,a ligand found on the surface of fungi,on gap junctional intercellular communication(GJIC)in cultured human corneal fibroblasts(HCFs).METHODS:Zymosan was added to the medium of cultured HCFs with or without the administration of mitogenactivated protein kinase(MAPK)inhibitors or the inhibitor kappa B kinase 2(IKK2)inhibitor IV.The protein and m RNA levels of connexin 43(Cx43)in HCFs were measured by Western blot,immunofluorescence,and quantitative reverse transcription-polymerase chain reaction(q RT-PCR)analyses.The GJIC activity was tested using a dye-coupling assay.RESULTS:The reduction of Cx43 protein and m RNA levels as well as a significant decrease in GJIC activity were observed in cultured HCFs when zymosan was added into the culture medium.Compared with controls(no zymosan),the protein level of Cx43 was reduced by 45%and 54%in the presence of zymosan at 200 and 600μg/m L,respectively(P<0.05);and it was reduced by 45%,48%,and 75%in the presence of zymosan(600μg/m L)for 24,36,and 48 h,respectively(P<0.05).The m RNA expression of Cx43 was reduced by 98%in the presence of zymosan(P<0.05).The effects of zymosan on Cx43 expression and GJIC activity were attenuated by the administration of PD98059[an extracellular signal-regulated kinase(ERK)signaling inhibitor](P<0.05),c-Jun NH2-terminal kinase(JNK)inhibitor II(P<0.05),and IKK2 inhibitor IV(P<0.05).CONCLUSION:Zymosan inhibits the activity of GJIC in cultured HCFs.This effect is likely regulated via the nuclear factor-κB(NF-κB),MAPK/ERK,and JNK signaling pathways.The inhibitory effects of zymosan on Cx43 expression and GJIC activity in HCFs may induce damage of corneal stroma during corneal fungal infection.

关键词： corneal fibroblast zymosan gap junction connexin 43 MAPKs