Protective effect of Liangxue Huayu decoction on human retinal pigment epithelial cell(ARPE-19)injury induced by hypoxia through autophagy pathway

Background:Exploring the protective mechanism of the Liangxue Huayu(LXHY)decoction on human retinal pigment epithelial(RPE)cells induced by hypoxia through the autophagy pathway.Methods:The appropriate LXHY decoction concentration was determined by CCK-8.ARPE-19 cells were divided into the normal control group(A group),CoCl_(2)group(B group),3-Methyladenine(3-MA)group(treated with 3-MA(the inhibition of autophagy pathway))(C group),blank serum(BS)group(D group),LXHY drug-contained serum(DCS)group(E group),and Rapamycin(RAP)group[treated with LXHY drug-contained serum combined with rapamycin group(the activation of autophagy pathway)](F group).Counting the number of autophagosomes and autolysosomes in each group of cells under transmission electron microscopy.After infection of cells in each group by mRFP-GFP-LC3 fusion protein adenovirus,the strength of autophagic flux was detected.The mRNA expression levels of LC3 and Beclin-1 were detected by Q-PCR.Results:CCK-8 assay results showed that LXHY DCS could inhibit the cell proliferation of ARPE-19 under hypoxia(all P<0.05).As the transmission electron microscopy assay result showed,compared with the normal control group,the number of autolysosomes was significantly increased in the CoCl_(2)group(P<0.05).Compared with CoCl_(2)group,the number of autolysosomes was significantly reduced the 3-Methyladenine group,blank serum group and LXHY drug-contained serum group(all P<0.001).As autophagic flux assay result showed,compared with the normal control group,the level of autophagosomes and autolysosomes were significantly risen in CoCl_(2)group(all P<0.001).Compared with the CoCl_(2)group,the level of autophagosomes and autolysosomes were significantly fell down in 3-Methyladenine group,blank serum group and LXHY drug-contained serum group(all P<0.05).The level of autolysosomes in the LXHY drug-contained serum group was lower than in the blank serum group(P<0.05).Compared with the LXHY drug-contained serum group,the levels of autophagosomes and autolysosomes were significantly risen in the LXHY drug-contained serum combined with the rapamycin group(all P<0.05).As the Q-PCR result showed,compared with the normal control group,the expression of LC3 and Beclin-1 mRNA were significantly reduced in the CoCl_(2)group(all P<0.001).Compared with the CoCl_(2)group,the expression of LC3 mRNA were significantly increased in the 3-Methyladenine group,blank serum group and LXHY drug-contained serum group(all P<0.001).Beclin-1 mRNA expression was increased significantly(all P<0.001)in the blank serum group and the LXHY drug-contained serum group.And Beclin-1 mRNA expression in the LXHY drug-contained serum group was statistically significant increased than blank serum group(P<0.001).In the LXHY drug-contained serum combined with the rapamycin group,the LC3 and Beclin-1 mRNA expression was reduced significantly compared with the LXHY drug-contained serum group(all P<0.001).Conclusion:The LXHY DCS has the ability to protect the human retinal pigment epithelial cell(ARPE-19)damage under hypoxia through the autophagy pathway.

Mitochondrial proteomic analysis of ecdysterone protection against oxidative damage in human lens epithelial cells

AIM: To investigate the protective effects of the natural medicinal monomer ecdysterone(ECR) with estrogenic activity against oxidative damage in human lens epithelial cells B3(HLE-B3) caused by hydrogen peroxide 21(H2 O2) and to pursue the possible mitochondrial proteomic regularity of the protective effects. · METHODS: HLE-B3 cells were treated with H2O2(300μmol/L),β-estuarial(E2; 10-8mol/L) and H2 O2,ECR(10-6mol/L) and H2 O2,or left untreated. Altered expression of all mitochondrial proteins was analyzed by protein array and surface-enhanced laser desorption ionization time of flight mass spectrometry(SELDI-TOF-MS). The mass/charge(M/Z) ratios of each peak were tested by the Kruskal-Wallis rank sum test,and the protein peak value of the M/Z ratio for each treatment by pair comparison was analyzed with the Nemenyi test. ·RESULTS: H2O2 up-regulated expression of two protein spots(with M/Z of 6 532 and 6 809). When E2 mitigated the oxidative damage,the expression of one protein spot(M/Z 6 532) was down-regulated. In contrast,ECR downregulated both of protein spots(M/Z 6 532 and 6 809). · CONCLUSION: ECR could effectively inhibite H2O2 induced oxidative damage in HLE-B3 cells. The protein spot at M/Z of 6 532 might be the target spot of ECR against oxidative damage induced by H2 O2.

Mechanism of Liangxue Huayu decoction in treatment of age-related macular degeneration based on network pharmacology and clinical validation

Background:This paper investigates the mechanism of Liangxue Huayu decoction treating age-related macular degeneration through network pharmacology and clinical verification.Methods:The potential targets of Liangxue Huayu decoction were retrieved by the BATMAN-TCM database,and age-related macular degeneration-related disease targets were collected using the OMIM,GeneCards,and CTD databases.The intersection of targets for Liangxue Huayu decoction for age-related macular degeneration was performed using the online tool Venny2.1.0.Protein-protein interaction network was drawn using the String database.And core targets were obtained with the CytoNCA plugin.The gene ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses were performed using the Metascape database.Finally,targets of Liangxue Huayu decoction for age-related macular degeneration were identified in clinical research.Results:In this study,832 potential action targets and 9,475 age-related macular degeneration-related disease targets were screened,and 575 intersection targets of Liangxue Huayu decoction and age-related macular degeneration were obtained.Protein-protein interaction network analysis showed that 15 core targets include chemokine 8,interleukin 1,histone deacetylase 2,estrogen receptor 2,and glutamate ionotropic receptor NMDA type subunit 1,et al.Gene ontology enrichment analysis included 2,727 biological processes,434 molecular functions,and 206 cellular components.Kyoto Encyclopedia of Genes and Genomes enrichment analysis selected 107 signaling pathways.Clinical validation showed that peripheral blood chemokine 8 and interleukin 1 levels were significantly lower after age-related macular degeneration patients compared with before treatment,and the difference was statistically significant(t chemokine 8=12.684,t interleukin 1=11.405,all P<0.001).Conclusion:By treating age-related macular degeneration with multiple targets and multiple pathways,Liangxue Huayu decoction can down-regulate the expression of inflammatory factors and treat age-related macular degeneration,which provides a basis for further studying the action mechanism of regulating age-related macular degeneration.

wARMD抗VEGF治疗应答不良的影响因素及靶向周细胞的治疗策略

随着人口老龄化程度的加剧,湿性年龄相关性黄斑变性(wARMD)的发病率逐渐上升,新生血管的形成导致黄斑区反复出血,是中老年人失明的主要原因之一。目前临床治疗以玻璃体腔注射抗血管内皮生长因子(VEGF)为一线治疗方法,但仍有部分患者对抗VEGF药物应答不良或无应答,导致临床治疗效果欠佳甚至无效,分析其具体影响因素将有利于指导临床制定方案。本文综述了高龄、疗程、注射次数、新生血管病灶特征、黄斑区结构、眼内房水细胞因子水平及遗传等因素对抗VEGF药物治疗应答的影响。此外,周细胞作为一种微血管壁的成分细胞,近年来发现其可影响抗VEGF药物治疗的敏感性,本文就周细胞在抗VEGF应答不良或不应答中的作用机制及靶向周细胞的解决策略研究现状进行综述。

OCTA定量分析补阳还五汤加味对非增殖型糖尿病视网膜病变患者视盘血流的影响

目的:观察补阳还五汤加味治疗气阴两虚、络脉瘀阻型非增殖型糖尿病视网膜病变(NPDR)的临床疗效,并应用光学相干断层扫描血管成像(OCTA)定量分析治疗前后视盘旁血流密度的变化。方法:采用前瞻性随机对照试验。收集2022-06/11期间我院收治的符合气阴两虚、络脉瘀阻型NPDR患者58例99眼,随机分为对照组29例48眼,采用2019年美国眼科学会发布的糖尿病视网膜病变推荐方案进行基础治疗(对于轻度NPDR患者进行控制血糖、糖尿病健康教育、定期随访;中度NPDR患者必要时增加局部或格栅样激光光凝治疗),观察组29例51眼在对照组的基础上加用补阳还五汤加味口服治疗1mo。比较两组患者治疗前后最佳矫正视力(BCVA)、中医证候疗效、视盘旁放射状毛细血管网血流密度(ppVD)及视盘旁视网膜神经纤维层(pRNFL)厚度变化。结果:治疗1mo后观察组BCVA(LogMAR)为0.20(0.10,0.30),较对照组0.30(0.20,0.40)明显改善(P<0.05)。治疗1mo后观察组中医证候疗效优于对照组(P<0.05)。观察组治疗1mo后各象限ppVD均较治疗前明显提高,且观察组各象限ppVD均高于对照组(P<0.05)。观察组治疗1mo后上方、颞侧象限及全周平均pRNFL厚度较治疗前升高,且观察组上方、颞侧、下方象限和全周平均pRNFL厚度均高于对照组(P<0.05)。结论:气阴两虚、络脉瘀阻型NPDR患者服用补阳还五汤加味后可改善视力、提高中医证候疗效,可能与其能提高视盘旁放射状毛细血管网血流密度和减少视盘旁神经纤维层的损伤有关。

跨上皮快速交联术联合术后配戴RGPCL治疗圆锥角膜的疗效

目的:探讨跨上皮快速交联术联合术后配戴硬性透气性角膜接触镜(RGPCL)治疗圆锥角膜患者的临床效果。方法:回顾性研究。选取2014-10/2015-10在福建中医药大学附属第二人民医院确诊为进展性圆锥角膜并进行跨上皮快速角膜交联术联合术后配戴RGPCL的患者16例25眼,平均年龄25.31±5.92岁,术前最大角膜曲率为58.96±6.76D,角膜最薄点厚度为439.52±53.24μm。使用核黄素及45mW/cm^(2)紫外光照射进行跨上皮快速交联,总能量7.2J/cm^(2),术后配戴RGPCL。术后随访3~6a,平均52.44±11.39mo,观察患者治疗前后BCVA(配戴RGPCL)、平坦子午线角膜曲率(K1)、陡峭子午线角膜曲率(K2)、最大角膜曲率(Kmax)、角膜屈光力(MPP)、角膜前后表面膨隆状态评价参数包括BCVf、BCVb、前表面曲率不对称指数(SIf)、后表面曲率不对称指数(SIb)、圆锥角膜前表面最高点(KVf)、圆锥角膜后表面最高点(KVb)、角膜最薄处厚度(ThkMin)、晶状体混浊、角膜内皮细胞计数。结果:所有患者手术过程顺利,无术中并发症,术后1d均有轻微刺痛伴异物感,轻微畏光流泪;2例2眼患者术后1d摘绷带镜后角膜上皮小片脱失,重新戴绷带镜3d后上皮愈合;随访期间所有患者均未出现晶状体混浊,角膜内皮细胞计数较术前无差异(P>0.05);术后BCVA(配戴RGPCL)较术前BCVA明显改善,术后Kmax、SIf、KVf、BCVf、BCVb均较术前降低(P<0.05)。结论:跨上皮快速角膜交联术联合RGPCL治疗圆锥角膜能有效且安全控制圆锥角膜进展,长期效果稳定。

Effects of Dendrobium candidum polysaccharides on microRNA-125b and mitogen-activated protein kinase signaling pathways in diabetic cataract rats

Background:Diabetic cataract is a common complication and a lens disorder in diabetes.Moreover,Dendrobium officinale polysaccharide(DOP)is found to alleviate diabetes complications.Consequently,microRNA-125b and mitogen-activated protein kinase signaling pathways play significant roles in diabetes.However,the mechanism and the effect of DOP on diabetic cataract remains unknown.Methods:Wistar male rats were randomly assigned to the control,model,and DOP groups.Diabetes was induced with streptozotocin.Furthermore,DOP was orally given for 12 weeks.The Lens Opacities Classification System III was used to evaluate lens opacity by slit-lamp microscope.The lens was then harvested for testing the mRNA expression of microRNA-125b,ERK1,ERK2,Raf and Ras using reverse transcription-polymerase chain reaction.The protein expression of ERK1,ERK2,Raf,and Ras was detected using the Western blot analysis.The targets of microRNA-125b were predicted in miRWalk 2.0.These targets were obtained from the Kyoto Encyclopedia of Genes and Genomes pathway.Results:The lens opacities of the rats in the control group were almost at C0N0.Moreover,the majority of the lens opacities in the model group were C3 to C4 and N1 to N2,and were mostly at C1 to C2 and N0 to N1 in the DOP group.The difference was statistically significant(P<0.05).Furthermore,the microRNA-125b expression in the model group is significantly higher compared with the control group.Conversely,the microRNA-125b expression in the DOP group is significantly decreased(all P<0.05).The mRNA expression of ERK1,ERK2,Raf,and Ras in the model groups were upregulated compared with those of the control group.However,the ERK1 and Raf mRNA expressions of the DOP group were lower compared with the model group(all P<0.05).The protein expression of ERK1,Raf,and Ras in the model group was significantly increased compared with those of the control group.The protein expression of ERK1,Raf,and Ras in the DOP group was significantly lower compared with the model group(all P<0.05).Moreover,3,378 genes of the microRNA-125b target were gained from the miRWalk.After Kyoto Encyclopedia of Genes and Genomes pathways analysis in Gene Set Enrichment Analysis,246 items were gained including Ras(rno04014)and mitogen-activated protein kinase(rno04010)signaling pathways.A positive correlation exists between microRNA-125b and mRNA expression of ERK1,ERK2,Raf and Ras(r=0.940,0.841,0.666,and 0.768;all P<0.05).Conclusion:DOP can alleviate the severity of the opacity of the lens in diabetic cataract rats via microRNA-125b and mitogen-activated protein kinase signaling pathways.Thus,microRNA-125b has something to do with the mitogen-activated protein kinase signaling pathway.

Analysis of the action mechanism of Si-Jun-Zi decoction in cataract by network pharmacology

Background:To explore the action mechanism of the classic ancient prescription Si-Jun-Zi decoction in slowing down cataract process by network pharmacology.Methods:In Traditional Chinese Medicines for Systems Pharmacology Database and Analysis Platform Database,screening the chemical components and functional targets of the classic ancient prescription Si-Jun-Zi decoction,and the targets of cataract disease were obtainded through GeneCards and the Online Mendelian Inheritance in Man Database.In the STRING database,the protein-protein interaction network was built.And network visualization were created in Cytoscape.The genetic ontology and Kyoto Gene and Genome Encyclopedia pathways of key proteins were enriched via the DAVID data platform.Results:Ninety-six active components and 33 key targets were obtained in this work.The protein-protein interaction network consisted of 33 proteins.There were 33 genetic ontology entries and 72 Kyoto Gene and Genome Encyclopedia pathway enrichment analysis.Conclusion:This work provided a new means to understand the pathogenesis of cataract and revealed the action mechanisms of the classic ancient prescription Si-Jun-Zi decoction.

Curcumin inhibits proliferation of human lens epithelial cells:a proteomic analysis

Objective: The incidence of after-cataracts [also known as posterior capsular opacification (PCO)] is between 30% and 50% three years following cataract surgery. Suppressing the proliferation of lens epithelial cells (LECs) is a primary goal in preventing PCO. Here, we investigated the proteomic regulation of the inhibitory effects of curcumin (Cur) on the proliferation of human lens epithelial B3 (HLE-B3) cells. Methods: Recombinant human basic fibroblast growth factor (rhbFGF) was used to induce proliferation of HLE-B3 cells, which were incubated with 20 mg/L Cur in a CO2 incubator for 24 h. Results: We found that the absorbance (A) value of rhbFGF group was significantly higher than the A value of the control group. Furthermore, the A value of the Cur group was significantly lower com- pared to the rhbFGF group, with an inhibition of 53.7%. Five different protein spots were obtained from proliferative HLE-B3 cells induced by rhbFGF. Eight different protein spots were obtained in HLE-B3 cells incubated with Cur. There were the common variational protein spots at mass/charge (m/z) ratios of 8 093 and 13 767 between rhbFGF group and control group as well as between the Cur group and rhbFGF group. Conclusions: These results show that Cur effectively inhibited HLE-B3 cell proliferation induced by rhbFGF. The protein spots at m/z of 8 093 and 13 767 may be the targets of Cur-induced inhibition of HLE-B3 cell proliferation. Cur may be a reliable and effective drug for pre- vention and treatment of polymerase chain reaction (PCR).

萘乙炔基封端的含硅聚酰亚胺的制备与性能

为了获得兼具良好热性能和加工性能的聚酰亚胺树脂,设计合成了不对称二胺(3-氨基-苯基)-(4'-氨基-苯基)-乙炔(AMPA),含萘环的封端剂3-(萘-1-乙炔基)苯胺(NAA)以及含硅二酐双(3,4-二羧基苯基)二甲基硅烷二酐.为研究结构与性能的关系,引入4,4'-双邻苯二甲酸酐(ODPA)和间氨基苯乙炔(APA)为对照二酐和封端剂,制备了一系列分子链中含硅和内炔基团的聚酰亚胺树脂PI-Si-Ⅰ(以APA为封端剂)和PI-Si-Ⅱ(以NAA为封端剂),以及与之相对照的树脂PI-O-Ⅰ和PI-O-Ⅱ(二酐单体为ODPA).PI-Si树脂在常见溶剂如四氢呋喃中具有很好的溶解度,而PI-Si-Ⅱ树脂更是具有低的熔体黏度和100℃宽的加工窗口.热失重的结果显示固化树脂具有良好的耐热性能,5 wt%热失重温度(Td5)在547℃左右,质量残留率在79%左右;热裂解分析结果表明在聚酰亚胺主链中引入的硅和内炔基团在高温环境中形成硅氧硅结构和苯环等刚性结构,从而提高树脂的耐热性.

Soluble Polyimide-reinforced TGDDM and DGEBA Epoxy Composites

Polyimide(PI)synthesized from aromatic diamine and dianhydrides via two-step poly-condensation method was highly soluble in TGDDM(MY-720)and DGEBA(E-51)at desirable temperature.TGDDM-PI(M-PI)and DGEBA-PI(E-PI)composites within 0.5%-3%PI loading could be prepared without organic solvent.On the cryogenically fractured surfaces of M-PI and E-PI composites,no obvious heterogeneous phase was observed by SEM.The mechanical properties were promoted significantly by PI,especially for impact strength.Adding 2%PI-2W into MY-720 and E-51 composites,the impact strength increased to 21 and 51 kJ/m2,the tensile strength increased by 62%and 19%,and the flexural strength by 18%and 13%,respectively,with slight increases in tensile modulus.These results were related to the promotion in plasticity of composites and changes in fragile→ductile fracture mode.Moreover,Tg and thermal stability of M-PI and E-PI were increased effectively.

Agglomeration behavior of petal-shaped cerium carbonate with different operating conditions

Agglomeration phenomenon can often be observed during the fabrication process of petal-shaped, nano-sized Ce2（CO3）3 powders. This kind of powder was produced by liquid-phase precipitation method with the template agent of polyallylamine hydrochloride （PAH）. In this study, the effects of the polymer concentration, pH value, and stirring speed on the agglomeration phe- nomenon of cerium carbonate were investigated. On the condition that the concentration of PAH is 0.9 g·L^-1, the pH value of the adjusting solution is 3-4, the final pH value is 8-10, and the stirring speed is 300-400 r·min^-1, it is found that the agglomeration phenomenon of Ce2（CO3）3 particles can be greatly prevented and the regular petal- shaped particles are formed.