Activated complement classical pathway in a murine model of oxygen-induced retinopathy

AIM: To investigate whether the complement system is involved in a murine model of oxygen-induced retinopathy(OIR).METHODS: Forty C57BL/6J newborn mice were divided randomly into OIR group and control group. OIR was induced by exposing mice to 75% ±2% oxygen from postnatal 7d(P7) to P12 and then recovered in room air.For the control group, the litters were raised in room air.At the postnatal 17d(P17), gene expressions of the complement components of the classical pathway(CP),the mannose-binding lectin(MBL) pathway and the alternative pathway(AP) in the retina were determined by quantitative real-time polymerase chain reaction(RT-PCR). Retinal protein expressions of the key components in the CP were examined by Western blotting.· RESULTS: Whole mounted retina in the OIR mice showed area of central hypoperfusion in both superficial and deep layers and neovascular tufts in the periphery.The expressions of C1 qb and C4 b genes in the OIR retina were significantly higher than those of the controls. The expression of retinal complement factor B(CFB) gene in OIR mice was significantly lower than those of the controls. However, the expressions of C3 and complement factor H(CFH) genes were higher. The protein synthesis of the key components involved in the CP(C1q, C4 and C3) were also significantly higher in OIR mouse retina. Although MBL-associated serine protease 1(MASP1) and MASP2 were detected in both the OIR and the control groups, the expressions were weak and the difference between the two groups was not significant.CONCLUSION: Our data suggest that the complement system CP is activated during the pathogenesis of murine model of OIR.

Complement factor Ⅰ knockdown inhibits colon cancer development by affecting Wnt/β-catenin/c-Myc signaling pathway and glycolysis

BACKGROUND Colon cancer(CC)occurrence and progression are considerably influenced by the tumor microenvironment.However,the exact underlying regulatory mechanisms remain unclear.AIM To investigate immune infiltration-related differentially expressed genes(DEGs)in CC and specifically explored the role and potential molecular mechanisms of complement factor I(CFI).METHODS Immune infiltration-associated DEGs were screened for CC using bioinformatics.Quantitative reverse transcription polymerase chain reaction was used to examine hub DEGs expression in the CC cell lines.Stable CFI-knockdown HT29 and HCT116 cell lines were constructed,and the diverse roles of CFI in vitro were assessed using CCK-8,5-ethynyl-2’-deoxyuridine,wound healing,and transwell assays.Hematoxylin and eosin staining and immunohistochemistry staining were employed to evaluate the influence of CFI on the tumorigenesis of CC xenograft models constructed using BALB/c male nude mice.Key proteins associated with glycolysis and the Wnt pathway were measured using western blotting.RESULTS Six key immune infiltration-related DEGs were screened,among which the expression of CFI,complement factor B,lymphoid enhancer binding factor 1,and SRY-related high-mobility-group box 4 was upregulated,whereas that of fatty acid-binding protein 1,and bone morphogenic protein-2 was downregulated.Furthermore,CFI could be used as a diagnostic biomarker for CC.Functionally,CFI silencing inhibited CC cell proliferation,migration,invasion,and tumor growth.Mechanistically,CFI knockdown downregulated the expression of key glycolysis-related proteins(glucose transporter type 1,hexokinase 2,lactate dehydrogenase A,and pyruvate kinase M2)and the Wnt pathway-related proteins(β-catenin and c-Myc).Further investigation indicated that CFI knockdown inhibited glycolysis in CC by blocking the Wnt/β-catenin/c-Myc pathway.CONCLUSION The findings of the present study demonstrate that CFI plays a crucial role in CC development by influencing glycolysis and the Wnt/β-catenin/c-Myc pathway,indicating that it could serve as a promising target for therapeutic intervention in CC.

Complement C3a activates osteoclasts by regulating the PI3K/PDK1/SGK3 pathway in patients with multiple myeloma

Objective:Myeloma bone disease(MBD)is the most common complication of multiple myeloma(MM).Our previous study showed that the serum levels of C3/C4 in MM patients were significantly positively correlated with the severity of bone disease.However,the mechanism of C3 a/C4 a in osteoclasts MM patients remains unclear.Methods:The formation and function of osteoclasts were analyzed after adding C3 a/C4 a in vitro.RNA-seq analysis was used to screen the potential pathways affecting osteoclasts,and the results were verified by Western blot,q RT-PCR,and pathway inhibitors.Results:The osteoclast area per view induced by 1μg/m L(mean±SD:50.828±12.984%)and 10μg/m L(53.663±12.685%)of C3 a was significantly increased compared to the control group(0μg/m L)(34.635±8.916%)(P<0.001 and P<0.001,respectively).The relative m RNA expressions of genes,OSCAR/TRAP/RANKL/cathepsin K,induced by 1μg/m L(median:5.041,3.726,1.638,and 4.752,respectively)and 10μg/m L(median:5.140,3.702,2.250,and 5.172,respectively)of C3 a was significantly increased compared to the control group(median:3.137,2.004,0.573,and 2.257,respectively)(1μg/m L P=0.001,P=0.003,P<0.001,and P=0.008,respectively;10μg/m L:P<0.001,P=0.019,P<0.001,and P=0.002,respectively).The absorption areas of the osteoclast resorption pits per view induced by 1μg/m L(mean±SD:51.464±11.983%)and 10μg/m L(50.219±12.067%)of C3 a was also significantly increased(33.845±8.331%)(P<0.001 and P<0.001,respectively)compared to the control.There was no difference between the C4 a and control groups.RNA-seq analysis showed that C3 a promoted the proliferation of osteoclasts using the phosphoinositide 3-kinase(PI3 K)signaling pathway.The relative expressions of PIK3 CA/phosphoinositide dependent kinase-1(PDK1)/serum and glucocorticoid inducible protein kinases(SGK3)genes and PI3 K/PDK1/p-SGK3 protein in the C3 a group were significantly higher than in the control group.The activation role of C3 a in osteoclasts of MM patients was reduced by the SGK inhibitor(EMD638683).Conclusions:C3 a activated osteoclasts by regulating the PI3 K/PDK1/SGK3 pathways in MM patients,which was reduced using a SGK inhibitor.Overall,our results identified potential therapeutic targets and strategies for MBD patients。

C3 Glomerulopathy and Therapeutic Potential of C5 Complement Inhibitors

C3 glomerulopathy is a disease including both dense deposit disease and C3 glomerulonephritis has an estimated prevalence of 2 to 3 per million. Originally, these pathologies were defined as glomerular pathology characterized by accumulation of C3 with absent or scanty immunoglobulin deposition. The keystone defect in both of these pathologies is the unregulated hyperactivity of alternative complement pathway. Specifically, in C3 glomerulopathy patients, there exists a prolongation of C3 cleavage which causes the uncontrolled alternative pathway activation. Many treatments have been investigated for treating C3 glomerulopathy to little or no avail, including calcineurin inhibitors, plasmapharesis, and anti-CD20 monoclonal antibodies. The next logical step is exploring the efficacy of anti-C5 monoclonal antibody therapy in C3 glomerulopathies to target the specific pathophysiology of this particular disease. Eculizumab is an anti-C5 monoclonal antibody that blocks the terminal step of complement activation. This drug has proven to be an effective treatment in other nephrologic pathologies that are caused by complement dysregulation. Here in this paper we discuss and present various case studies and clinical trials available that experiment with Eculizumab in patients with either dense deposit disease or C3 glomerulonephritis. In most of these patients, treatment with Eculizumab has demonstrated clinical and biochemical improvements in kidney function. These results provide encouraging evidence that suggest Eculizumab as a promising therapy for patients with C3 glomerulopathy and warrant that more extensive clinical trials can be designed as a next step.

IgA肾病患者补体经典途径在血液及尿液中的活化及与肾损伤的关系

目的 研究循环系统中经典途径补体活化及调节方式在IgA肾病（IgAN）中的致病作用及与肾损伤的关系.方法 IgA肾病组30例,10例狼疮性肾炎（LN）患者作阳性对照,30例健康体检者作对照组.采用ELISA试剂盒检测IgA肾病患者和对照组血及尿液中经典途径补体激活标志物C1q、经典途径调节因子（sCR）1,分析补体浓度与肾组织病理结果及临床生化指标的相关性.将IgA肾病组患者按照病理Lee氏结果进行分组,Lee氏Ⅰ～Ⅲ级定为轻度损伤组,Lee氏Ⅳ～Ⅴ级定为重度损伤组,比较两组间的补体浓度差异.分析血C1q与sCR1间相关性.结果 IgA肾病组患者血清C1q及sCR1水平均高于对照组（P＜0.05）,而IgA肾病组与LN组间比较差异无统计学意义（P ＞0.05）;LN组患者尿液C1q浓度明显高于另外2组（P＜0.01）.当血清肌酐＞ 133μmol/L时,血C1q与肌酐水平呈显著负相关（P＜0.05）.尿C1q与肌酐水平呈显著正相关（P＜0.05）.Lee氏Ⅳ～Ⅴ级组患者血液及尿液C1q均明显低于Lee氏Ⅰ～Ⅲ级组（P＜0.05）.血sCR1与血C1q间呈显著正相关（P＜0.05）.结论 IgA肾病患者循环系统中可能存在补体经典途径激活通路,尤其是在肾损伤严重组,且与疾病的严重程度相关.IgA肾病中可溶型CR1对C1q存在介导调节作用.

一种新型的定量测定特异性体液免疫功能的溶血空斑试验

根据补体经典途径激活时，可通过“旁立损伤”机制使邻近正常细胞溶解破坏的原理，该种溶血空斑试验不仅可以用来定量检测分泌ＩｇＭ的抗体形成细胞，也可直接用来定量检测分泌ＩｇＧ的抗体形成细胞。与传统溶血空斑试验相比，其最大的优点是可用来检测机体对任何一种可溶性抗原的特异性体液免疫应答能力，从而扩展了应用范围。

三条补体通路在IgA肾病血液中的活化及致病作用

目的:研究不同的补体活化类型在Ig A肾病(Ig A nephropathy,Ig AN)中的活化及其致病作用,拓展Ig AN的发病机制理论。方法:设立Ig AN疾病组30例,疾病对照组狼疮性肾炎(Lupus nephritis,LN)患者8例作阳性对照,本院查体中心筛选健康人30例作正常对照。采用商品化的ELISA试剂盒检测Ig AN患者及对照者血清中三条补体激活渠道的标志性成分:三条补体活化通路中的共同裂解产物C3a、C5a,及溶解型终末期膜攻击复合物s C5b-9,经典途径标志物C1q,凝集素途径标志物L-ficolin(FCN2),经典途径及凝集素途径激活后的共同标志物C4d,替代途径标志物补体Bb因子。分析血清中的补体浓度与临床生化指标的相关性。结果:所有因子三组间的比较均有显著差异(P<0.05)。补体因子与临床指标间存在不同程度的相关性,其中Bb、C3a等补体裂解产物与血肌酐间呈正相关关系,FCN2与尿红细胞计数间呈负相关关系,而C1q与临床指标间无相关性。结论:Ig AN患者血液中或许存在补体经典途径、凝集素途径及替代途径的活化,并且补体激活参与临床及肾脏病理损伤,补体激活程度影响临床及病理损伤程度。

人精浆免疫抑制因子SF_1对血清补体的影响

SF_1是正常人精浆经Sephadex G 100分离后得到的第一组分。将正常人血清和杂种小鼠混合血清作为补体来源,每份血清均以SF_1处理为实验组,并设不加SF_1的自身对照组,从下列实验中分别观察SF_1对补体系统的影响:1.CH_(50)试验证明,SF_1能抑制人血清中总补体活性(P<0.01);2.SF_1能抑制人血清中补体的溶血作用,并随血清中加入SF_1剂量的增加而抑制作用增强;3.利用YC 花环试验证实SF_1可抑制补体替代途径的激活;4.SF_1不影响用单向琼脂扩散试验测定中C_3、C_4的量,证明SF_1没有影响C_3、C_4的抗原性。上述实验结果提示:SF_1在体外实验中能抑制补体的活性,表现于对补体经典途径和替代途径激活的抑制。

补体对大肠杆菌的致死过程

实验证明，在不依赖特异性机体的条件下，在Ｅ．ｃｏｌｉＢ接触补体后的溶菌反应中，先释放碱性磷酸酶，后释放β－半乳糖昔酶，在一段时间内体外两种酶与溶菌率成正相关性，说明该情况下补体最初作用部位是细胞壁，但致死效应则涉及对细胞膜的损伤。

Zadeh模糊集合理论存在问题证明及其改进——一个满足全部经典集合公式的C-模糊集合系统

Zadeh模糊集合理论具有不能正确描绘客观世界的全部模糊现象,特别是不能描绘相交而不“包含或者分散包含”的情况,不可能存在反集等两个严重缺点;定义了不存在的反集这一严重错误,导致了思维、逻辑和概念混乱.但是,Zadeh等把错误缺点说成为“对传统的挑战”、“摆脱传统的约束”[2-序]的先进成果.企图用“算子”拼盘(不是像概率论那样各种公式有统一的解释)来掩盖缺点,导致了系统混乱(不清楚什么时候需要使用什么算子),误导人们以为模糊集合理论必然与常规思维、逻辑和概念相悖.为此,分析和证明了Zadeh模糊集合的错误.介绍了一个新模糊集合系统:C-模糊集合系统,它能克服Zadeh模糊集合理论的全部错误和缺点,能正确地描绘客观世界的全部模糊现象,有反集.它是经典集合系统的特例而不是推广,能满足全部经典集合的公式,与正常思维、逻辑和概念一致.

High-throughput RNA sequencing reveals the anti-inflammatory mechanism of baicalin on Propionibacterium acnes-induced acne in rabbits

Objective:Propionibacterium acnes(P.acnes)plays an important role in the development of acne,an inflammatory skin disease with a high-incidence.In this study,we used high-throughput RNA sequencing(RNA-seq)to reveal the anti-inflammatory mechanism of baicalin on P.acnes-induced acne in rabbits.Methods:The Kligman method was used to induce acne in the ears of New Zealand rabbits.The effect of baicalin on the acne model was evaluated by the number of acne lesions and hematoxylin and eosin(H&E)staining of acne tissues.Enzyme-linked immunoabsorbent assay was used to measure the protein expression levels of tumor necrosis factor a(TNFA),interleukin-1 b(IL1B),IL6,and IL8 in the serum of rabbits.RNA-seq was performed to investigate the mechanism of anti-inflammatory activities of baicalin on acne.Immunohistochemical analysis and Western blot were used to validate the expression levels of related proteins in acne tissues.Results:Baicalin treatment significantly reduced the number of acne lesions and lesions of the ear as well as levels of serum inflammatory cytokines.RNA-seq data showed that baicalin treatment globally suppressed inflammation,especially the TNF signaling pathway and Staphylococcus aureus infection pathway,in the rabbit acne model.Conclusion: Baicalin effectively ameliorates P. acnes-induced acne in rabbits by suppressingthe inflammatory response in rabbits.

Reclassification of membranoproliferative glomerulonephritis:Identification of a new GN:C3GN

This review revises the reclassification of the mem-branoproliferative glomerulonephritis （MPGN） after the consensus conference that by 2015 reclassified all the glomerulonephritis basing on etiology and patho-genesis, instead of the histomorphological aspects. After reclassification, two types of MPGN are to date recognized： The immunocomplexes mediated MPGN and the complement mediated MPGN. The latter type is more extensively described in the review either because several of these entities are completely new or because the improved knowledge of the complement cascade allowed for new diagnostic and therapeutic approaches. Overall the complement mediated MPGN are related to acquired or genetic cause. The presence of circulating auto antibodies is the principal acquired cause. Genetic wide association studies and family studies allowed to recognize genetic mutations of different types as causes of the complement dysregulation. The complement cascade is a complex phenomenon and activating factors and regulating factors should be distinguished. Genetic mutations causing abnormalities either in activating or in regulating factors have been described. The diagnosis of the complement mediated MPGN requires a complete study of all these different complement factors. As a consequence, new therapeutic approaches are becoming available. Indeed, in addition to a nonspecifc treatment and to the immunosuppression that has the aim to block the auto antibodies production, the specific inhibition of complement activation is relatively new and may act either blocking the C5 convertase or the C3 convertase. The drugs acting on C3 convertase are still in different phases of clinical development and might represent drugs for the future. Overall the authors consider that one of the principal problems in fnding new types of drugs are both the rarity of the disease and the consequent poor interest in the marketing and the lack of large international cooperative studies.

大肠杆菌不依赖特异性抗体激活补体系统经典途径的研究

除已被公认的两条补体激活途径，即经典途径和替代途径外，还存在着一条不依赖抗原──抗体复合物而激活补体的经典途径。用Ｅ．ｃｏｌｉＢ、Ｅ．ｃｏｌｉＫ１２ｇａｌ＋的细胞壁加入到补体参与的溶血实验中发现有明显的溶血抑制作用，从而说明大肠杆菌的细胞壁对不依赖抗体而激活补体经典途径有密切关系。

中国传统哲学思想视阈下的古诗英译管窥

关于中国古典诗歌的翻译方法,一直以来争执不断,各抒己见。本文梳理了中国传统哲学思想对汉语言的浸润和对译者翻译观的影响,分析了不同地域译者的译文,提出自己的观点,即鉴于中国古典诗歌的独特性,独特的翻译方法具有可行性。一言以蔽之,异化翻译于中西方文化互补有着不可替代的作用。

据居延汉简讨论汉代动补式问题

汉语动补式起源的时间,主要有“先秦说”、“两汉说”和“魏晋说”,汉语史界尚难定论。以20世纪30年代以来出土的居延汉简资料居延汉简出现了动词前带有副词的动补式,出现了“V”、“VC”紧邻的情况,出现多例进入“为…所…”被动句的动补式,传世文献稀见的“敝尽”、“折伤”、“攻得”、“击伤”,在居延汉简中都有较多例子。因此,动补式在西汉时代确已出现,并有较多的用例。

血清补体系统在流行性出血热患者中的变化及其意义

应用单向免疫扩散法测定了流行性出血热(EHF)患者的7种补体成份和血浆素原(Pg).结果表明,CT脂酶抑制剂在少尿期、多尿期和恢复期均高于正常(P<0.01);补体C_1q在多尿期高于正常(P<0.01);补体C_4在发热期和少尿期低于正常(P<0.01),以后逐渐恢复正常;补体C_3,C_5和C_9的变化与C_4相似;B因子在少尿期低于正常(P<0.01),在多尿期高于正常(P<0.05);Pg始终高于正常水平但在少尿期有回降.说明EHF患者不仅有补体经典途径的识别阶段、活化阶段和膜攻击阶段的变化,而且亦有旁路激活,其活化程度与病情有关.

浅谈中国传统园林之人文内涵

中国古典园林是中国传统文化的一部分 .以儒家思想为主体的文化形态在中国传统园林中得以充分的体现 ,它包括天人合一的自然观 ,礼乐相成的伦理道德观以及多元互融 。

人补体杀伤小鼠N9小胶质细胞及亚溶破模型的建立

目的:探讨正常人血清(norm al hum an serum,NHS)补体杀伤小鼠N9小胶质细胞的机制,建立人补体攻膜复合物(sub lytic m embrane attack comp lex,sMAC)N9细胞亚溶破刺激模型。方法:用阻断补体活化途径的方法探讨N9细胞活化人补体系统的机制;采用微量补体反应性溶破法,以酵母多糖(Zymosan,Z)活化急性期病人血清制备补体优球蛋白C56,EDTA-NHS作为C7-C9的来源,组装N9细胞sMAC亚溶破模型;CCK-8比色实验确定sMAC亚溶破剂量;激光共聚焦鉴定sMAC沉积;脱落细胞计数及台盼蓝拒染法分别判定细胞黏附力变化及脱落细胞活力。结果:未致敏N9细胞可通过旁路途径直接活化人血清补体系统;当C56稀释度在1∶500以上,NHS(含1 mmol/L EDTA)稀释度为1∶20时,膜攻击复合物(m embrane attack comp lex,MAC)活性逐渐降低,细胞溶破减少,确定C56 1∶500,NHS 1∶20为N9细胞亚溶破补体量;激光共聚焦鉴定sMAC沉积于N9细胞表面;亚溶破剂量MAC刺激N9细胞后与对照组相比细胞脱落增加(P<0.05),而细胞活力正常。结论:探讨了小鼠N9细胞活化人补体的机制;通过建立N9细胞人补体sMAC亚溶破模型,证实sMAC刺激N9细胞后可降低细胞黏附力但不影响细胞活力;为sMAC对中枢神经系统小胶质细胞亚溶破刺激效应的深入研究提供了理论与实验基础。

应用酵母双杂交方法筛选与猪瘟病毒N^(pro)蛋白相互作用的宿主蛋白

为研究猪瘟病毒(CSFV)与宿主细胞间的相互作用,本研究采用酵母双杂交方法以猪瘟病毒Npro蛋白为诱饵,从猪外周血单个核细胞(PBMC)cDNA表达文库中筛选与之相互作用的蛋白,经共转化试验表明,共筛选到12个与Npro蛋白相互作用的宿主蛋白,应用Cytoscape 2.8.2软件绘制了蛋白质间相互作用关系图,根据GO分析结果,这些蛋白分别参与细胞代谢、细胞生长、免疫等过程,经双分子荧光互补(BiFC)试验表明筛得的部分宿主蛋白与CSFV Npro具有相互作用。本研究中筛选得到的蛋白质对CSFV复制的影响有待进一步深入研究。

几项补体测定指标间相关性分析

对６８例志愿者与患者作了血清总补体溶血活性（ＣＨ_（５０））、旁路途径溶血活性（ＡＰＨ_（５０））、Ｃ_４活性与含量、Ｂ因子（ＢＦ）的活性与含量、Ｃ_３含量测定。活性测定用免疫溶血法，含量测定用单扩散法。结果表明，经典途径指标中Ｃ_４活性检出补体水平异常的阳性率为最高；旁路途径指标中则以ＢＦ活性阳性率最高。ＣＨ_（５０）与Ｃ_４活性、ＡＰＨ_（５０）与ＢＦ活性均极显著相关（Ｐ＜０．０１），ＣＨ_（５０）与ＡＰＨ_（５０）、Ｃ_４含量与Ｃ_３含量显著相关（Ｐ＜０．０５），其余两两指标间无明显相关关系（Ｐ＞０．０５）。作者认为，Ｃ_４和ＢＦ活性可作为检测补体激活的两项基本的过筛试验，单扩散法作补体成份定量不反映补体活性水平。