PRRSV degrades MDA5 via dual autophagy receptors P62 and CCT2 to evade antiviral innate immunity

Porcine reproductive and respiratory syndrome virus(PRRSV)is a major economically devastating pathogen that has evolved various strategies to evade innate immunity.Downregulation of antiviral interferon largely promotes PRRSV immunoevasion by utilizing cytoplasmic melanoma differentiation-associated gene 5(MDA5),a receptor that senses viral RNA.In this study,the downregulated transcription and expression levels of porcine MDA5 in PRRSV infection were observed,and the detailed mechanisms were explored.We found that the interaction between P62 and MDA5 is enhanced due to two factors:the phosphorylation modification of the autophagic receptor P62 by the upregulated kinase CK2αand the K63 ubiquitination of porcine MDA5 catalyzed by the E3 ubiquitinase TRIM21 in PRRSV-infected cells.As a result of these modifications,the classic P62-mediated autophagy is triggered.Additionally,porcine MDA5 interacts with the chaperonin containing TCP1 subunit 2(CCT2),which is enhanced by PRRSV nsp3.This interaction promotes the aggregate formation and autophagic clearance of MDA5-CCT2-nsp3 independently of ubiquitination.In summary,enhanced MDA5 degradation occurs in PRRSV infection via two autophagic pathways:the binding of MDA5 with the autophagy receptor P62 and the aggrephagy receptor CCT2,leading to intense innate immune suppression.The research reveals a novel mechanism of immune evasion in PRRSV infection and provides fundamental insights for the development of new vaccines or therapeutic strategies.

Repeatability and reproducibility of corneal higher-order aberrations measurements after small incision lenticule extraction using the Scheimpfug-Placido topographer

Background:To evaluate the precision of corneal higher-order aberrations measurements after small incision lenticule extraction(SMILE)using the Sirius Scheimpfug-Placido topographer(CSO,Italy).Methods:Seventy-fve eyes from 75 postoperative subjects were included in this prospective study.Three consecutive corneal aberrometric measurements were obtained with the Scheimpfug-Placido topographer by two experienced operators to assess intra-and inter-observer reproducibility.The within-subject standard deviation(Sw),test-retest repeatability(TRT)and the intraclass correlation coefcient(ICC)were calculated.Results:For intraobserver repeatability of anterior and total corneal aberrations,all ICCs were more than 0.922,except for trefoil(0.722 to 0.768).The ICCs of total root mean square(RMS),coma Z(3,±1),and spherical aberration Z(4,0)were over 0.810 while higher-order RMS,trefoil Z(3,±3),and astigmatism II Z(4,±2)were below 0.634 for posterior corneal surface aberrations.All Sw values for all types of aberrations were equal to or below 0.07μm.Regarding interobserver reproducibility,all TRT values were no more than 0.12μm,0.05μm,and 0.11μm for anterior,posterior,and total corneal aberrations,respectively.The ICC values ranged from 0.875 to 0.989,from 0.686 to 0.976 and over 0.834 for anterior,posterior,and total corneal aberrations,respectively.Conclusions:The repeatability of measurements of anterior and total corneal aberrations with the Sirius system in corneas after SMILE surgery was high,except for trefoil.There was some variability in posterior corneal aberrometric measurements.High reproducibility of corneal aberrometric measurements was observed between measurements of both examiners,except for trefoil,with poor to moderate reproducibility.

Sirius Scheimpflug-Placido versus. ultrasound pachymetry for central corneal thickness: meta-analysis

Background:To compare the difference in central corneal thickness(CCT)measurements in normal eyes between a rotating Scheimpflug camera combined with a Placido-disk corneal topographer(Sirius,CSO,Italy)and ultrasound pachymetry(USP).Methods:A systematic literature search was conducted for relevant studies published on PubMed,Medline,EMBASE,and the Cochrane Library and ClinicalTrials.gov from inception to August 1st,2019.Primary outcome measures were CCT measurements between Sirius and USP.A random effects model was used to pool CCT measurements.Results:A total of twelve studies involving 862 eyes were included in this meta-analysis.The meta-analysis found CCT measurements between Sirius and USP to be statistically significantly different(P<0.0001).The mean difference between Sirius and USP was-11.26μm with a 95%confidence interval(CI)(-16.92μm,-5.60μm).The heterogeneity Was IP=60%(P=0.004).Conclusion:CCT measurements with the Sirius Scheimpflug-Placido topographer were statistically significantly lower than USP.However,it may be argued that the mean difference of 11.26μm is not a clinically significant difference.

重视自适应光学OCT的工程技术研究及转化应用

自适应光学相干断层扫描(AO-OCT)在2003年首次报道并取得了巨大技术进步,较传统OCT具有更好的成像能力,能获得接近人眼衍射极限的眼底视网膜微结构的高分辨率立体图像,为眼科领域提供了一种新的诊断和研究工具。笔者就AO-OCT的主要工程技术研究及转化应用做一评述,以期加深对AO-OCT重要性的认识。

国人版角膜接触镜配戴者生活质量量表的研制与评估

目的:研制国人版角膜接触镜配戴者生活质量量表(CLIQ)并评估其信度和效度,分析得分的影响因素。方法:采用横断面研究。对英国原版CLIQ进行翻译、回译、文化调试,形成国人版量表。选择2015年4─12月在温州医科大学附属眼视光医院就诊的角膜接触镜配戴者205例进行问卷调查,其中再随机选取20例由2名调查员分别进行调查,对问卷数据进行统计分析。结果:回收有效问卷201例。视功能维度因应答率小于50%被删除。量表Cronbach'sα系数为0.77,眼部症状维度为0.68,便利性、经济和情感维度均>0.7,量表Guttmann Split-Half系数为0.89,眼部症状、便利性、经济和情感维度分别为0.75、0.78、0.65和0.93。认知维度Cronbach'sα系数和Guttmann Split-Half系数均为0.49故被删除;删除条目20后健康关注维度Cronbach'sα系数和Guttmann Split-Half系数分别为0.68和0.73。2名调查员之间的评分结果高度相关(r=0.87~0.99,P<0.001)。各条目的条目水平内容效度指数为0.83~1.00,量表的平均水平内容效度指数为0.95。探索性因子分析提取的5个因子分别对应各维度,累计方差贡献率60.43%。眼部症状维度与年龄(r=-0.153,P=0.03)和戴镜年数(r=-0.167,P=0.018)轻度相关,情感维度与戴镜年数轻度相关(r=0.209,P=0.003)。量表总分在不同戴镜年数[(3.60±0.34)年vs.(3.71±0.32)年,P=0.026]、每月戴镜天数[(3.60±0.33)d vs.(3.70±0.33)d,P=0.041]和无/有眼部症状(3.71±0.32 vs.3.60±0.34,P=0.021)的配戴者间差异有统计学意义。软镜配戴者的经济维度得分高于硬镜配戴者(t=2.588,P=0.017)。结论:删去视功能维度、认知维度和条目20后,CLIQ量表的信度和效度较好,戴镜年数越长、每月戴镜天数越多以及无眼部症状的患者得分较高,硬镜配戴者在花费方面的担忧更多。

A comprehensive Chinese experience against SARS-CoV-2 in ophthalmology

The 2019 novel coronavirus disease(COVID-19)has now swept through the continents and poses a global threat to public health.Several investigations have been conducted to identify whether COVID-19 can be transmitted through the ocular route,and the conclusion is that it is a potential route but remains uncertain.Due to the faceto-face communication with patients,frequent exposure to tears and ocular discharge,and the unavoidable use of equipment which requires close proximity,ophthalmologists carry a high risk of contracting severe acute respiratory syndrome coronavirus 2(SARS-CoV-2).Based on 33 articles published by Chinese scholars,guidelines and clinical practice experience in domestic hospitals,we have summarized the Chinese experience through the lens of ophthalmology,hoping to make a contribution to protecting ophthalmologists and patients around the world.

Porcine Immunoglobulin Fc Fused P30/P54 Protein of African Swine Fever Virus Displaying on Surface of S. cerevisiae Elicit Strong Antibody Production in Swine

African swine fever virus(ASFV)infects domestic pigs and European wild boars with strong,hemorrhagic and high mortality.The primary cellular targets of ASFV is the porcine macrophages.Up to now,no commercial vaccine or effective treatment available to control the disease.In this study,three recombinant Saccharomyces cerevisiae(S.cerevisiae)strains expressing fused ASFV proteins-porcine Ig heavy chains were constructed and the immunogenicity of the S.cerevisiae-vectored cocktail ASFV feeding vaccine was further evaluated.To be specific,the P30-Fcγand P54-Fcαfusion proteins displaying on surface of S.cerevisiae cells were produced by fusing the Fc fragment of porcine immunoglobulin IgG1 or IgA1 with p30 or p54 gene of ASFV respectively.The recombinant P30-Fcγand P54-Fcαfusion proteins expressed by S.cerevisiae were verified by Western blotting,flow cytometry and immunofluorescence assay.Porcine immunoglobulin Fc fragment fused P30/P54 proteins elicited P30/P54-specific antibody production and induced higher mucosal immunity in swine.The absorption and phagocytosis of recombinant S.cerevisiae strains in IPEC-J2 cells or porcine alveolar macrophage(PAM)cells were significantly enhanced,too.Here,we introduce a kind of cheap and safe oral S.cerevisiae-vectored vaccine,which could activate the specific mucosal immunity for controlling ASFV infection.

Porcine FcεRI Mediates Porcine Reproductive and Respiratory Syndrome Virus Multiplication and Regulates the Inflammatory Reaction

Porcine reproductive and respiratory syndrome virus(PRRSV) shows characteristic antibody-dependent enhancement(ADE) of infection and causes porcine systemic inflammation, which is similar to a type I allergic reaction; however, the role of porcine FceεRI in ADE is still unclear. In this study, the expression of different Fc receptors(Fc Rs) on macrophages was investigated in a PRRSV 3D4/21 cell infection model in the presence or absence of PRRSV antibody. The transcription level of Fcc II and FceεRI was significantly up-regulated under PRRSV-antibody complex infection. Internalization and proliferation of PRRSV were promoted by the ADE mechanism when FceεRI was expressed in permissive 3D4/21 cells and the non-permissive cell line HEK 293T. Transcriptome sequencing data showed that the expression levels of AKT,ERK and other signal molecules in the anti-inflammatory pathway were significantly increased, especially in the cells infected with the PRRSV-antibody immune complex. Inflammatory regulatory molecules such as PLA2G6, LOX, TRPM8 and TRPM4 were significantly up-regulated following PRRSV infection but significantly down-regulated in the cells infected with the PRRSV-antibody immune complex. Our results demonstrated that FceεRI could be involved in PRRSV ADE, the antigen presenting process and regulation of the inflammatory response during PRRSV infection, which provides new insights into PRRSV infection mediated by FceεRI and the PRRSV-antibody immune complex.

Catquest 9SF-CN量表在白内障中的最小临床重要差异研究

目的:评估Catquest 9SF-CN量表应用于白内障手术人群的反应度,并结合校标法和分布法确定最小临床重要差异值(MCID)。方法:前瞻性队列研究。对纳入温州医科大学附属眼视光医院的213例白内障术前患者进行Catquest 9SF-CN量表调查。术后1~3个月对来院复查的患者再次进行量表调查和7个选项的校标条目调查。分析量表的天花板和地板效应,通过配对t检验分析手术前后量表得分和视力的差异,采用效应值、标准化反应均数和变化率3个指标联合评估量表的反应度。然后根据术前视力、术前并发症(有/无)和术眼数量(单眼/双眼)分组,计算手术带来的得分变化。通过单因素方差分析(根据术前视力分组)或独立样本t检验(根据并发症或者术眼数量分组)分析组间差异。分布法通过1个效应值、1个测量标准误和1/2个得分变化标准差估算MCID,校标法通过校标选项和得分变化的线性回归分析估算MCID,最后根据平均值法确定取值。结果:共144例完成Catquest 9SF-CN量表的随访调查。量表术前的天花板和地板效应为4.2%,术后提高至17.4%。手术带来的量表总得分变化为11.0±7.9,logMAR视力变化为0.4±0.4,两者有相关性(r=0.30,P<0.001),且各自手术前后的差异具有统计学意义(t视力=13.44,t得分=16.75,P<0.001)。量表的效应值、标准化反应均数和变化率分别是1.71、1.40、44.46%。有完整随访资料的患者根据术前视力分成3组,或者根据术眼数量分成2组后,组间的得分变化差异均无统计学意义。根据并发症分组后,差异有统计学意义(t=2.90,P=0.004)。分布法中1个效应值、1个测量标准误和1/2个得分变化标准差估算MCID分别为1.71、2.12、3.96分。校标法中量表得分变化和校标条目选项呈负相关(r=-0.35,P<0.001),回归公式中的β系数为3.74,用来估计MCID值。结合分布法和校标法取平均值得到最终MCID值2.88分。结论:Catquest 9SF-CN量表适用于白内障患者生活质量评估,在中国白内障患者手术前后的反应度良好,并建立了MCID值2.88分作为应用参考。

The C/EBPβ-Dependent Induction of TFDP2 Facilitates Porcine Reproductive and Respiratory Syndrome Virus Proliferation

Porcine reproductive and respiratory syndrome(PRRS) is an important infectious disease caused by porcine reproductive and respiratory syndrome virus(PRRSV), leading to significant economic losses in swine industry worldwide. Although several studies have shown that PRRSV can affect the cell cycle of infected cells, it is still unclear how it manipulates the cell cycle to facilitate its proliferation. In this study, we analyzed the mRNA expression profiles of transcription factors in PRRSV-infected 3D4/21 cells by RNA-sequencing. The result shows that the expression of transcription factor DP2(TFDP2) is remarkably upregulated in PRRSV-infected cells. Further studies show that TFDP2 contributes to PRRSV proliferation and the PRRSV nucleocapsid(N) protein induces TFDP2 expression by activating C/EBPb. TFDP2 positively regulates cyclin A expression and triggers a less proportion of cells in the S phase, which contributes to PRRSV proliferation. This study proposes a novel mechanism by which PRRSV utilizes host protein to regulate the cell cycle to favor its infection. Findings from this study will help us for a better understanding of PRRSV pathogenesis.

Sirius Scheimpflug–Placido versus ultrasound pachymetry for central corneal thickness:meta-analysis

Background:To compare the difference in central corneal thickness(CCT)measurements in normal eyes between a rotating Scheimpflug camera combined with a Placido-disk corneal topographer(Sirius,CSO,Italy)and ultrasound pachymetry(USP).Methods:A systematic literature search was conducted for relevant studies published on PubMed,Medline,EMBASE,and the Cochrane Library and ClinicalTrials.gov from inception to August 1st,2019.Primary outcome measures were CCT measurements between Sirius and USP.A random effects model was used to pool CCT measurements.Results:A total of twelve studies involving 862 eyes were included in this meta-analysis.The meta-analysis found CCT measurements between Sirius and USP to be statistically significantly different(P<0.0001).The mean difference between Sirius and USP was−11.26μm with a 95%confidence interval(CI)(−16.92μm,−5.60μm).The heterogeneity was I^(2)=60%(P=0.004).Conclusion:CCT measurements with the Sirius Scheimpflug-Placido topographer were statistically significantly lower than USP.However,it may be argued that the mean difference of 11.26μm is not a clinically significant difference.

基于BiblioShiny的先天性白内障可视化文献计量分析(2011—2020年)

目的:基于BiblioShiny程序分析2011—2020年全球先天性白内障的研究现状与发展趋势,为进一步的研究提供参考信息。方法:文献计量分析方法。运用基于R语言的科学文献计量BiblioShiny程序,对2011—2020年Web of ScienceTM核心合集数据库(WoSCC)收录的关于先天性白内障的相关文献,从期刊、国家与作者分布、共词分析、共被引分析、合作网络等深入分析全球先天性白内障的研究知识基础和研究前沿热点。结果:①共纳入1492篇研究文献,以中国和美国发文量最多。②《Molecular Vision》杂志的出版物发表量排名榜首,刊文74篇,其次是《PLoS One》,刊文66篇,二者均为美国期刊。③发文量第1位的是来自加利福尼亚州帕罗奥多市斯坦福大学医学院眼科的Lambert SR博士。④先天性白内障领域的知识基础是先天性白内障和基因突变,目前的学术研究热点是基因表达主题。⑤研究方向可以分为3大集群:第1集群为先天性白内障的发病率、危险因素和手术治疗等;第2集群的研究内容主要是分子遗传、基因突变和蛋白质功能改变等;第3集群的研究内容主要集中在晶状体蛋白的结构、稳定性和功能等。结论:2011—2020年先天性白内障研究领域中,中国和美国为发文量最多的国家,但是美国在国家间、机构间的合作网络均远大于中国,提示中国可提高国家、机构间的合作率。目前先天性白内障主流研究方向可分为临床治疗、基因层面和蛋白质层面。

The precision and agreement of corneal thickness and keratometry measurements with SS-OCT versus Scheimpflug imaging

Purpose:To assess the repeatability and reproducibility of swept-source optical coherence tomography(SS-OCT)and Scheimpflug system and evaluate the agreement between the two systems in measuring multiple corneal regions in children.Methods:Pachymetric and keratometric maps for both systems were evaluated.Central,midperipheral and peripheral corneal thickness(CT),keratometry and astigmatism power vectors were recorded.The three outcomes yielded by the same observer were used to assess intraobserver repeatability.The differences in the mean values provided by each observer were used to evaluate interobserver reproducibility.Within-subject standard deviation,test-retest repeatability(TRT)and coefficient of variation(CoV)were used to analyze the intraobserver repeatability and interobserver reproducibility.Paired T-test and Bland-Altman were used to appraise interdevice agreement.Results:Seventy-eight eyes of 78 children were included.The CoV was≤2.12 and 1.10%,respectively,for repeatability and reproducibility.TRT and CoV were lower for central and paracentral CT measurements than for peripheral measurements.The SS-OCT device generated higher precision when acquiring CT data,whereas Scheimpflug system showed higher reliability when measuring corneal keratometry.Although the CT readings measured using SS-OCT were significantly thinner than Scheimpflug device(P<0.001),the central and thinnest CT values were still of high agreement.The interdevice agreement of keratometry measurement was high for the central corneal region and moderate for the paracentral and peripheral areas.Conclusions:The precision of CT measurements by SS-OCT was higher,while the reliability of keratometry measurements by the Scheimpflug system was higher in children.Apart from the measured values in the central corneal region,the thickness and keratometry readings should not be considered interchangeable between the two systems.

Effect of orthokeratology on precision and agreement assessment of a new sweptsource optical coherence tomography biometer

Background:To evaluate the effect of orthokeratology on precision of measurements in children using a new swept-source optical coherence tomography(SS-OCT)optical biometer(OA-2000),and agreement between its measurements and those provided by the commonly used IOLMaster based on partial coherence interferometry(PCI).Methods:This study recruited fifty-one eyes of 51 normal children(8–16 years).An operator took measurements with the two biometers.Then,a second operator took measurements with the SS-OCT biometer.After orthokeratology was performed for one month,the same operators repeated the same procedures.Axial length(AL),mean keratometry(Km)at 2.5 mm and 3.0 mm diameters(Km2.5 and Km3.0),central corneal thickness(CCT),anterior chamber depth(ACD),lens thickness(LT)and corneal diameter(CD)were analyzed.Results:With the SS-OCT optical biometer,the test-retest repeatability of AL measurements was<0.06 mm.For all parameters,the coefficients of variation were<1.23%and the intraclass correlation coefficients were>0.95.The 95%limits of agreement of difference between the two devices for CD parameter were up to 1.53 mm.After orthokeratology,the fluctuation ranges of difference for Km3.0 measurement was 1.11 times higher than before orthokeratology,while the absolute values of difference for AL,Km2.5,ACD and CD measurements were comparable.Conclusions:Before and after orthokeratology,the SS-OCT biometer showed high repeatability and reproducibility for all measurements.Wearing orthokeratology contact lenses affected the agreement between SS-OCT and PCI biometers for Km3.0 measurements.The CD measurement showed poor agreement between the two devices.

High intercorneal symmetry in corneal biomechanical metrics

Backgroud:To evaluate the symmetry of corneal biomechanical metrics,measured using an ocular response analyzer(ORA)and self-built corneal inflation test platform,in bilateral rabbit corneas and to investigate their relationship with physical intraocular pressure(IOPp).Methods:Twenty fresh enucleated eyes from ten rabbits were used for ex vivo whole ocular globe inflation.IOP was increased from 7.5 to 37.5 mmHg with 7.5 mmHg steps and biomechanical metrics were acquired using the ORA.At least 3 examinations were performed at each pressure stage.Two biomechanical metrics,corneal hysteresis(CH)and corneal resistance factor(CRF)were recorded and analyzed as a function of IOPp.Corneal specimens were then excised from the intact ocular globe and tested under inflation conditions up to 45.7 mmHg posterior pressure.The experimental pressure-deformation data was analyzed using an inverse modeling procedure to derive the stress-strain behavior of the cornea.Results:A comparison of corneal shape parameters showed no statistically significant difference(P>0.05)between bilateral eyes.Similarly,there were no statistically significant differences in values of CH,CRF and corneal stiffness(as measured by the tangent modulus,Et)between bilateral eyes(CH:F=0.94,P=0.54;CRF:F=4.42,P=0.35;Et:F=3.15,P=0.12)at different pressure levels.IOPp was highly correlated with CRF while the relationship with CH was less pronounced.Conclusions:An obvious interocular symmetry in biomechanical metrics is found in this research.IOP has been shown to have important influences on the value of CRF provided by ORA.

The precision and agreement of corneal thickness and keratometry measurements with SS-OCT versus Scheimpflug imaging

Purpose:To assess the repeatability and reproducibility of swept-source optical coherence tomography(SS-OCT)and Scheimpflug system and evaluate the agreement between the two systems in measuring multiple corneal regions in children.Methods:Pachymetric and keratometric maps for both systems were evaluated.Central,midperipheral and peripheral corneal thickness(CT),keratometry and astigmatism power vectors were recorded.The three outcomes yielded by the same observer were used to assess intraobserver repeatability.The differences in the mean values provided by each observer were used to evaluate interobserver reproducibility.Within-subject standard deviation,test-retest repeatability(TRT)and coefficient of variation(CoV)were used to analyze the intraobserver repeatability and interobserver reproducibility.Paired T-test and Bland-Altman were used to appraise interdevice agreement.Results:Seventy-eight eyes of 78 children were included.The CoV was≤2.12 and 1.10%,respectively,for repeatability and reproducibility.TRT and CoV were lower for central and paracentral CT measurements than for peripheral measurements.The SS-OCT device generated higher precision when acquiring CT data,whereas Scheimpflug system showed higher reliability when measuring corneal keratometry.Although the CT readings measured using SS-OCT were significantly thinner than Scheimpflug device(P<0.001),the central and thinnest CT values were still of high agreement.The interdevice agreement of keratometry measurement was high for the central corneal region and moderate for the paracentral and peripheral areas.Conclusions:The precision of CT measurements by SS-OCT was higher,while the reliability of keratometry measurements by the Scheimpflug system was higher in children.Apart from the measured values in the central corneal region,the thickness and keratometry readings should not be considered interchangeable between the two systems.

角膜胶原交联中核黄素浓度检测方法的研究进展

角膜胶原交联术(CXL)可以用于治疗进展期圆锥角膜、感染性角膜炎、角膜融解和角膜基质水肿等病变。临床上常用的CXL方案为核黄素-紫外光交联,角膜中的核黄素浓度和分布是影响疗效和安全性的重要因素之一。目前对角膜中核黄素浓度进行检测的方法主要包括荧光显微镜成像、高效液相色谱法、分光光度法、无创实时检测、眼前节裂隙灯显微镜观察法以及基质辅助激光解吸/电离质谱成像技术(MALDI-IMS)等,不同方法之间结果可能存在差异。现对目前研究中的核黄素浓度及其检测方法做一概述。