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Nano Research 2018年第10期11卷 5566-5572页

作者： Wenjing Qi Yufei Zhang Zdravko Kochovski Jue Wang Yan Lu Guosong Chen Ming Jiang The State Key Laboratory of Molecular Engineering of Polymers and Department of Macromolecular Science Fudan University Shanghai 200438 China Soft Matter and Functional Materials Helmholtz-Zentrum Berlin for Materialien und Energie Berlin 14109 Germany Institute of Chemistry University of Potsdam 14476 Potsdam Germany

Recently, we proposed a new strategy to construct artificial plant protein assemblies, which were induced by adding a small molecule, based on dual supramolecular interactions. In this paper, we further explored this method by employing Human galectin-1 （Gal-1） as a building block to form self-assembled microribbons. Two non-covalent interactions, including lactose-lectin binding and dimerization of Rhodamine B （RhB）, induced by the small molecule ligand addition, were involved in the crosslinking of the animal protein, resulting in the formation of assemblies. By using transmission electron microscopy （TEM）, cryo-electron microscopy （cryo-EM）, and three-dimensional （3D） tomographic analysis, we arrived at a possible mechanistic model for the microribbon formation. Furthermore, the morphology of protein assemblies could be fine-tuned by varying the incubation time, the protein/ligand ratio, and the chemical structures of ligands. Interestingly, the formation of protein microribbons successfully inhibited Gal-1 induced T-cell agglutination and apoptosis. This is because the multivalent and dynamic interactions in protein assemblies compete with the binding between Gal-1 and the glycans on cell surfaces, which suppresses the function of Gal-1 in promotion of tumor progression and metastasis.

关键词： protein self-assembly supramolecularinteractions galectin cell agglutination

galectin-3与CDC25B在胃癌中的表达及意义

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中华医学遗传学杂志 2009年第3期26卷 288-292页

作者：张秀明姚根有张布衣王玲玲赵敏浙江大学医学院病理学与法医学研究所杭州310058 浙江中医药大学附属新华医院杭州市迪安医学检验中心

目的探讨半乳糖凝集素3mRNA（galectin-3）和细胞分裂周期蛋白25BmRNA（celldivisioncycle25B，CDC25B）在胃癌中的表达及其与临床病理参数及预后的关系。方法应用组织微阵列技术和原位杂交法检测220例胃癌组织和31份正常胃黏膜组织中Ga... 详细信息

目的探讨半乳糖凝集素3mRNA（galectin-3）和细胞分裂周期蛋白25BmRNA（celldivisioncycle25B，CDC25B）在胃癌中的表达及其与临床病理参数及预后的关系。方法应用组织微阵列技术和原位杂交法检测220例胃癌组织和31份正常胃黏膜组织中galectin3mRNA和CDC25BmRNA的表达情况。结果220例胃癌中galectin3tuRNA和CDC25BtuRNA的阳性表达率分别为58．6％、54．1％；galectin-3mRNA表达与肿瘤大小、TNM（tumor，lymphnode，metastasis）分期、浸润深度、脉管侵犯、淋巴结转移及远处转移呈正相关；CDC25BmRNA表达也与肿瘤大小、TNM分期、脉管侵犯、淋巴结转移及远处转移呈正相关；galectin3mRNA表达与CDC25BmRNA表达呈正相关；galectin-3mRNA和CDC25BmRNA阳性表达病例的平均生存时间和5年生存率均明显低于阴性表达的病例。结论galectin3和CDC25B基因的表达促进了胃癌的发生和发展，是指导临床治疗及评估预后的有效指标。

关键词：胃癌组织微阵列 galectin 3基因 CDC25B基因原位杂交

聚乙二醇干扰素α-2a对慢性乙型肝炎患者外周血单个核细胞galectin-3表达的影响

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中华肝脏病杂志 2014年第5期22卷 336-339页

作者：李宜余龙吴晓玲安徽省立医院感染科合肥230001 安徽省六安市人民医院感染科

目的研究聚乙二醇干扰素α-2a（Peg—IFNα-2a）治疗慢性乙型肝炎（CHB）患者的应答情况与外周血单个核细胞（PBMC）galectin-3表达的关系，探讨galectin-3对Peg-IFN α-Za抗病毒疗效的影响。方法治疗组CHB患者接受Pe扩IFN α-2a治疗48... 详细信息

目的研究聚乙二醇干扰素α-2a（Peg—IFNα-2a）治疗慢性乙型肝炎（CHB）患者的应答情况与外周血单个核细胞（PBMC）galectin-3表达的关系，探讨galectin-3对Peg-IFN α-Za抗病毒疗效的影响。方法治疗组CHB患者接受Pe扩IFN α-2a治疗48周，随访24周，根据血清学以及病毒学检测结果，分为治疗24周有效组、治疗48周有效组及治疗无效组。PCR和Westernblot方法检测治疗前、治疗24周时及治疗48周时患者PBMCgalectin-3基因和蛋白质表达情况。根据数据类型分别采用t检验或χ^2检验进行数据比较。结果Peg-IFN α-2a治疗有效组患者PBMC galectin-3的基因及蛋白质相对表达量明显高于正常对照组和治疗无效组（P值均〈0．05）；治疗24周有效组galectin-3的基因及蛋白质相对表达量在24周和48周时均明显高于正常对照组（P值均〈0．05）。治疗48周有效组在48周时的galectin-3基因和蛋白质相对表达量明显高于治疗24周时（P〈0．05）。结论Peg-IFN α-2a可能通过上调人体外周血单核细胞galectin-3表达发挥抗病毒作用。

关键词：肝炎乙型慢性干扰素α-2a 外周血单个核细胞 galectin

galectin Hco-gal-m from Haemonchus contortus modulates goat monocytes and T cell function in different patterns

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Galectin Hco-gal-m from Haemonchus contortus modulates goat ...

中国畜牧兽医学会兽医寄生虫学分会第十三次学术研讨会

作者：王望 Shuai Wang Hui Zhang Cheng Yuan RuoFeng Yan XiaoKai Song LiXin Xu 李祥瑞南京农业大学动物医学院

Background:Monocytes and T cells are two major subpopulations of peripheral blood mononuclear cells(PBMC) and play an essential role in the innate and adaptive immune *** members of the galectin family show multiple and distinct regulatory effects on different cell *** studies have demonstrated that the galectin from Haemonchus contortus(Hco-gal-m) performed immunomodulatory effects on goat PBMC,however,which subpopulation of PBMC is the primary target of Hco-gal-m and whether the immune modulations share the same mechanism remain ***:In this study,the developmental expression of Hco-gal-m was analyzed by RT-PCR and Western blot *** distribution of Hco-gal-m in adult worm was detected by an immunohistochemical *** bmding activity of the recombinant Hco-gal-m(rHco-gal-m) on goat monocytes and T cells were assessed by flow *** immunomodulatory effects of Hco-gal-m on cytokine secretion,cell activation and apoptosis were observed by co-incubation of rHco-gal-m with goat monocytes and T ***:Hco-gal-m was expressed in L4 as well as adult worms and predominantly localized at the internal surface of the worm ***-gal-m could bind to both monocytes and T *** engagement of rHco-gal-m decreased the production of IL-6,IL-10 and TNF-α in T cells,however,it significantly increased the secretion of IL-10 in *** rHco-gal-m exposure,the expression of MHC-Ⅱ on monocytes and that of CD25 on T cells were ***,T cell proliferations were potently inhibited by *** addition,rHco-gal-m induced apoptosis in T cells,but not significantly in ***:Our results indicated that rHco-gal-m modulated goat monocytes and T cell function in different patterns.

关键词： Haemonchus contortus galectin PBMC Monocytes T cells

A novel fungal galectin induces apoptosis through inhibition of farnesyltransferase activity

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A novel fungal galectin induces apoptosis through inhibition...

湖北省生物工程学会2004年年会

作者： Xin Tong ChenGuang Zhao Yi Liang Wei Wei Hui Sun Yipeng Qi Key laboratory of Virology, Ministry of Education, College of Life Science, Wuhan University, Wuhan 430072, Hubei Province, People’s Republic of China

Inhibitors of farnesyltransferase (FTase) are potential therapeutic agents for Ras-related human cancer,We have previously identified galectin from fungi,named AAL(Agrocybe aegerita lectin) which displayed potent tumor-suppressing function. Here we clone the cDNA of demonstrate a functional interaction between gelectin and FTase. whose farnesylation activity is essential for the membrane targeting of Ras oncogenes. The inhibitory function of gelectin on FTase leads to retention of Ras to the cvtosol, and

关键词： galectin farnesyltransferase. apoptosis. Agrocybe aegerita fungal lectin Fas Ras. quantitative real-time PCR flow cytometry.

Analysis of the up-regulated expression genes in hemocytes of pearl oyster（Pinctada fucata,Gould 1850） responding to bacterial stimulation:Identification of a galectin gene

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Analysis of the up-regulated expression genes in hemocytes o...

2008年中国水产学会学术年会

作者： Zhongliang Wang~(a b c) Zaohe Wu~(* a b c) Jichang Jian~(a b c) Yishan Lu~(a b c) a College of Fisheries.Guangdong Ocean University Zhanjiang China b Guangdong Provincial Key Laboratory of Pathogenic Biology and Epidemiology for Aquatic Economic Animals Zhanjiang.China c Guangdong Key Laboratory of Control for Diseases of Aquatic Economic Animals Zhanjiang China

＜正＞The pearl oyster,Pinctada fucata,which is the most important species cultured for production of marine pearls in China and has become an important asset to the economy, frequently suffered mass mortalities in recent ***,almost nothing is known on the molecular basis of the pearl oysters immune *** order to identify immune-related genes in hemocytes from the pearl oysters stimulated with Vibrio alginolyticus,a forward suppression subtractive hybridization（SSH） cDNA library was constructed and the up-regulated genes were analyzed.A total of 600 clones were randomly selected and sequenced. The sequence analysis showed that 167 clones were matched with sequences in the Genbank database which represented 66 different genes with known functions,among the known genes, 7 genes were possibly related to immune ***,which plays crucial roles in the innate immunity systems of vertebrates and invertebrates,was identified and characterized for the first time in the pearl oyster in this paper,and the full-length of galectin cDNA was 1344bp, consisting of a 5’-terminal untranslated region（UTR） of 26 bp,a 3’ terminal UTR of 313 bp, and an open reading frame（ORF） of 1005 bp encoding a polypeptide of 334 amino acids with a theoretical molecular weight of 36.76 kDa and an estimated isoelectric point of 8.27. Fluorescent real-time quantitative RT-PCR was used to examine the expression of galectin gene in haemocytes of *** after the challenge of bacteria Vibrio *** was a clear time-dependent expression pattern of galectin after bacterial *** results obtained in this work could be the first step leading to the understanding of molecular mechanism by which the *** respond to pathogens.

关键词： SSH Immune genes galectin Bacterial challenge Real-time PCR

Transmembrane protein 147(TMEM147):another partner protein of Haemonchus contortus galectin on the goat PBMC

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Transmembrane protein 147(TMEM147):another partner protein o...

中国动物学会寄生虫学专业委员会第十届全国寄生虫学青年工作者学术研讨会

作者： Yan Li Chen Yuan Ruofeng Yan Xiaokai Song Lixin Xu Xiangrui Li College of Veterinary Medicine Nanjing Agricultural University

Recombinant galectins of male and female Haemonchus contortus(r Hco-gal-m/f) have been recognized as significant regulators of the functions of goat peripheral blood mononuclear cells(PBMC). In previous research, transmembrane protein 63A(TMEM63A) was identified as a partner protein in the regulation associated with H. contortus infection. However, in the identification of binding partners for galectins of male and female H. contortus(Hco-gal-m/f) by yeast two-hybrid(YTH) screening, it was found that the transmembrane protein 147(TMEM147) could also bind to Hco-gal-m/f. In this study, the functions of TMEM147 in the regulations of H. contortus galectin on the goat PBMC were investigated. To identify Hco-gal-m/f-interacting proteins, a yeast two-hybrid system to detect interactions was used. Co-immunoprecipitation and immunoblotting were used to validate the interaction between recombinant galectins of male H. contortus(r Hco-gal-m) and candidate binding protein. The localization of TMEM147 in PBMC was explored by immunofluorescence in confocal imaging studies. Flow cytometry was used to determine the distribution of TMEM147 in T cells, B cells and monocytes in PBMC. The modulatory effects of rH co-gal-m and TMEM147 on cell proliferation, phagocytosis, nitric oxide production, migration, apoptosis and cytokine mRNA transcription were observed by co-incubation of r Hco-gal-m and knockdown of the tmem147 gene. In this research, it was demonstrated that TMEM147 could bind to r Hco-gal-m/f. Immunofluorescence assays showed that TMEM147 was localized to the cell membrane and within the cell membrane in goat PBMC. Flow cytometric analysis revealed that TMEM147 was expressed in all B cells and monocytes in goat PBMC. However, 3.8 % of T cells did not express this protein. Knockdown of the tmem147 gene using RNA interference(RNAi) showed that the interaction of galectin with TMEM147 mainly mediated cell proliferation, cell apoptosis, transcription of interleukin-10(IL-10) an

关键词： galectin Goat PBMC Haemonchus contortus Member receptor TMEM147

Transmembrane protein 63A is a partner protein of Haemonchus contortus galectin in the regulation of goat peripheral blood mononuclear cells

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Transmembrane protein 63A is a partner protein of Haemonchus...

中国畜牧兽医学会兽医寄生虫学分会第十三次学术研讨会

作者：袁橙 Hui Zhang Wang Wang Yan Li Ruofeng Yan Lixin Xu Xiaokai Song 李祥瑞南京农业大学动物医学院

Background:Hco-gal-m and-f were two isoforms of galectin cloned from male and female Haemonchus contortus,respectively,and it was demonstrated that recombinant Hco-gal-m and-f could act as immune ***,little is known about the receptors or binding partners of these galectins in the *** research of the molecular mechanisms that govern the interactions between these galectins and host molecules will fill a gap in our understanding how parasite galectins interact with host ***:A yeast two-hybrid system was used to identify the binding partners of Hco-gal-m and-f in this *** interaction between rHco-gal-m and candidate binding protein was validated by *** localization of transmembrane protein 63A(TMEM63A) in peripheral blood mononuclear cells(PBMCs)was detected by *** distribution of TMEM63 A in T cells,B cells and monocytes in PBMCs was detected by flow *** immunomodulatory effects of Hco-gal-m and TMEM63 A on cell proliferation,migration,apoptosis,nitric oxide production and cytokine secretion were observed by co-incubation of rHco-gal-m and TMEM63A-siRNA with goat PBMCs and ***:We found that TMEM63 A,a functionally unknown protein,from goat PBMCs could bind to Hco-gal-m *** showed that TMEM63 A was localized to the cell *** cytometric analysis revealed that TMEM63 A was expressed in the majority of goat *** using RNA interference to knockdown expression of TMEM63 A,the PBMC proliferation and migration were significantly increased,while the influence of rHco-gal-m on monocyte phagocytosis,PBMC nitric oxide production and migration were potently *** addition,the production of IL-10,IFN-γ and TGF-β induced by rHco-gal-m were also ***:Our results show that TMEM63 A is a binding partner of Hco-gal-m/f,and involved in the immune responses of host PBMCs induced by Hco-gal-m for the first time.

关键词： galectin Haemonchus contortus Partner protein TMEM63A

捻转血矛线虫galectin C端CRD差异氨基酸突变对其生物学活性的影响

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捻转血矛线虫Galectin C端CRD差异氨基酸突变对其生物学活性的影响

作者：王婧南京农业大学

学位级别：硕士

捻转血矛线虫半乳糖结合凝集素(galectin)含有串联重复的两个糖结合域(carbohydrate recognition domain，CRD)。雌、雄成虫重组galectin在C端CRD基序中存在一个差异氨基酸，雌虫中该基序是WGNEGR，而雄虫中该基序为WGNEER。这两种galec... 详细信息

捻转血矛线虫半乳糖结合凝集素(galectin)含有串联重复的两个糖结合域(carbohydrate recognition domain，CRD)。雌、雄成虫重组galectin在C端CRD基序中存在一个差异氨基酸，雌虫中该基序是WGNEGR，而雄虫中该基序为WGNEER。这两种galectin在血凝活性、糖结合能力、体外抑制山羊外周血淋巴细胞细胞因子mRNA转录以及体外诱导山羊外周血淋巴细胞凋亡等方面均显示出一定的差异。基于以前的研究，本论文主要进行了以下工作： 1．雌、雄捻转血矛线虫galectins C端Clid差异氨基酸的突变及其糖结合域的克隆通过定点突变技术更换Hco-gal-m与Hco-gal-f C端CRD的差异氨基酸。Hco-gal-mC端CRD(wGNEER)中的第二个E被突变为G，得到与HCO-gal-f相同的突变galectin(MG mutate);Hco-gal-f C端CRD(WGNEGR)中的G被突变为E，得到与Hco-gal-m相同的突变galecthl(FE mutate)。DNA序列分析证明突变体碱基序列与设计序列完全一致。同时通过PCR分别克隆了Hco-gal-f／m 340bp左右的N端CRD(FNh、MNh)和540bp左右C端CRD(FCh、MCh)，DNA序列分析证明碱基序列完全正确。 2．重组galectins的原核表达及表达产物的纯化利用温度敏感原核表达系统在大肠杆菌中表达了重组MG mutate、FE mutate、MNh、MCh、FNh、FCh。SDS-PAGE分析发现重组MG mutate和FE mutate的分子量大小为32．5kDa左右、重组MNh和FNh为14.4kDa左右、重组MCh和FCh为20.0kDa左右。上述重组galectins以包涵体形式存在。用8mol／L尿素溶解包涵体使之变性后在含有递减浓度尿素的TE缓冲液中分步透析复性，可得到浓度较高重组galectins。 3．捻转血矛线虫galectins C端CRD差异氨基酸突变对其血凝活性的影响通过比较Hco-GAL-f、Hco-GAL-m以及它们的突变体MG mutate、FE mutate对人B型红细胞的血凝活性来检测C端CRD中的第二个谷氨酸(E)对捻转血矛线虫血凝活性的影响。血凝试验发现，C端CRD中第二个氨基酸E的改变颠倒了重组galectins凝集人B型红细胞的能力。纯化重组Hco-GAL-m和Hco-GAL-f的最高凝集效价分别为2和2，而它们的突变体MG mutate和FE mutate，则分别是2和2。此结果说明，捻转血矛线虫C端CRD中第二个氨基酸E的改变影响了重组galectins的血凝活性。同时通过比较捻转血矛线虫galectins N端CRDs和C端CRDs的血凝活性发现，MCh和FCh的最高凝集效价分别为2和2，而MNh和FNh则分别为2和2，揭示四个重组CRD中C端CRDs凝集人B型红细胞的能力强于N端CRDs。糖抑制试验结果表明，乳糖和半乳糖都能有效抑制上述重组galectins的糖结合能力，乳糖最低抑制浓度低于半乳糖最低抑制浓度。 4．捻转血矛线虫galectins C端CRD差异氨基酸突变对其糖结合能力的影响用亲和层析方法比较了重组Hco-GAL-f、Hco-GAL-m、MG mutate、FE mutate、MCh、FCh、MNh、FNh的糖结合能力，以此来检测C端CRD中的第二个谷氨酸(E)对捻转血矛线虫重组galectin糖结合能力的影响以及两端CRD糖结合能力的差异。比较亲和层析图谱发现Hco-GAL-m和FE mutate(Hco-GAL-f的突变体)能有效的结合于a-乳糖层析介质，而Hco-GAL-f、MG mutate(Hco-GAL-m的突变体)则不能与a-乳糖层析介质结合。MCh和FCh与a-乳糖层析介质的结合能力与Hco-GAL-m和FEmutate相比大大减弱，但仍存在;MNh和FNh则不能与a-乳糖层析介质结合。此结果表明，捻转血矛线虫C端CRD中第二个氨基酸E的改变影响了重组galectins的糖结合能力，并且C端CRDs的糖结合能力强于N端CRDs。 5．捻转血矛线虫galectins诱导山羊淋巴细胞凋亡的活性部位的研究为了探讨重组的雄、雌捻转血矛线虫galectin(Hco-GAL-m／f)诱导天然宿主山羊外周血淋巴细胞凋亡的具体活性部位，采用DNA琼脂糖凝胶电泳、流式细胞仪分析的方法，检测了重组Hco-GAL-f、Hco-GAL-m、MG mutate、FE mutate、MCh、FCh、MNh和FNh对淋巴细胞凋亡的影响。结果显示，重组Hco-GAL-m、Hco-GAL-f、MGmutate、FE mutate及四个CRD

关键词：捻转血矛线虫 galectin 血凝活性糖结合能力细胞凋亡

中药周期疗法调控反复种植失败患者子宫内膜容受性及对galectin的影响

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中药周期疗法调控反复种植失败患者子宫内膜容受性及对Galectin的...

作者：曹媛媛山东中医药大学

学位级别：硕士

目的:对行体外受精-胚胎移植(In vitro fertilization-embryo transfer,IVF-ET)反复种植失败(recurrent implantation failure,RIF)的患者给予中药周期疗法干预治疗,设置对照组进行对比。通过研究两组患者中医肾虚临床症状、体征及实验... 详细信息

目的:对行体外受精-胚胎移植(In vitro fertilization-embryo transfer,IVF-ET)反复种植失败(recurrent implantation failure,RIF)的患者给予中药周期疗法干预治疗,设置对照组进行对比。通过研究两组患者中医肾虚临床症状、体征及实验室指标,研究两组患者子宫内膜容受性的特征性标记分子半乳糖凝集素(galectin)的表达,分析中药周期疗法对子宫内膜容受性的作用机制及对胚胎种植率和临床妊娠率的影响。方法:研究对象为120例行冻融胚胎移植(frozen-thawed embryo transfer,FET)的患者,其中RIF患者60例为观察组,输卵管因素行IVF-ET的患者60例为对照组。观察组给予中药周期疗法干预一个疗程,对照组不予干预。比较患者中医肾虚临床症状体征及实验室指标;比较排卵日及排卵后6日子宫内膜容受性特征性标记分子半乳糖凝集素(galectin)的表达;比较两组患者FET周期胚胎种植率及临床妊娠率。结果:1.观察组治疗后肾虚症状及体征较前改善,与对照组比较无统计学差异(P>0.05);2.观察组治疗后血清基础内分泌水平较前改善,与对照组比较无统计学差异(P>0.05);3.观察组治疗后排卵日及排卵后6日血清galectin水平升高、子宫内膜galectin及m RNA表达均有提高,排卵日子宫内膜厚度、分型及内膜下血流均较前改善,与对照组比较无统计学差异(P>0.05);4.观察组治疗后FET周期移植日及FET后14日血清激素水平,胚胎种植率及临床妊娠率,与对照组比较无统计学差异(P>0.05)。结论:中药周期疗法能改善RIF患者中医肾虚证候及基础内分泌环境,改善排卵日子宫内膜容受性相关指标,提高了FET周期子宫内膜容受性特征性标记分子galectin的表达,从而提高了子宫内膜容受性与胚胎种植率及临床妊娠率。

关键词：中药周期疗法反复种植失败子宫内膜容受性 galectin