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Poly(A)-specific ribonuclease protein promotes the proliferation,invasion and migration of esophageal cancer cells
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作者 Fu-Wei Zhang Xiao-Wei Xie +5 位作者 Meng-Hua Chen Jian Tong Qun-Qing Chen Jing Feng Feng-Ti Chen Wen-Qi Liu 《World Journal of Gastroenterology》 SCIE CAS 2023年第31期4783-4796,共14页
BACKGROUND Bioinformatics analysis showed that the expression of the poly(A)-specific ribonuclease(PARN)gene in gastric cancer,head and neck squamous cell carcinoma,melanoma,cervical cancer and lung squamous cell carc... BACKGROUND Bioinformatics analysis showed that the expression of the poly(A)-specific ribonuclease(PARN)gene in gastric cancer,head and neck squamous cell carcinoma,melanoma,cervical cancer and lung squamous cell carcinoma tissues was significantly higher than that in normal tissues and was associated with high stage and poor prognosis.The expression of the PARN gene in esophageal cancer(EC)tissue is also significantly higher than that in normal tissues,but the effect of PARN on the proliferation,migration and invasion of EC cells remains unclear.AIM To investigate the relationship between PARN and the proliferation,migration and invasion of EC cells.METHODS The EC tissues of 91 patients after EC surgery and 63 paired precancerous healthy tissues were collected.PARN mRNA levels were measured using a tissue microarray,and the PARN expression level was evaluated using immunohistochemistry to analyze the relationship between PARN expression and clinicopathologic features as well as the survival and prognosis of patients.In addition,the effects of PARN gene knockout on tumor cell proliferation,invasion and migration were studied by using shRNA during the in vitro culture of EC cell lines Eca-109 and TE-1,and the effects of the PARN gene on tumor growth in vivo were verified by a xenotransplantation nude mice model.RESULTS The expression of PARN in EC tissues was higher than that in adjacent normal tissues,and the level of PARN expression was significantly positively correlated with lymphatic metastasis.Patients with high PARN levels had poor overall survival.BIM,IGFBP-5 and p21 levels were significantly increased in the PARN knockout group,while the expression levels of the antiapoptotic proteins Survivin and sTNF-R1 were significantly decreased in the apoptotic antibody array data.In addition,the expression levels of Akt,p-Akt,PIK3CA and CCND1 in the downstream signaling pathway regulating EC progression were significantly decreased.The culture of EC cell lines confirmed that the apoptosis rate of EC cells was significantly increased,the growth and proliferation of tumor cells were significantly inhibited,and the invasion and migration ability of tumor cells were significantly decreased after PARN gene knockout.In vivo experiments of BALB/c nude mice transfected with Eca-109 cells expressing control shRNA(sh-NC)and PARN shRNA(sh-PARN)showed that the tumor volume and weight of nude mice treated with sh-PARN were significantly decreased compared with those of nude mice treated with sh-NC,indicating that PARN knockdown significantly inhibited tumor growth in vivo.CONCLUSION PARN has antiapoptotic effects on EC cells and promotes their proliferation,invasion and migration,which is associated with the development of EC and poor patient prognosis.PARN may become a potential target for the diagnosis,prognosis prediction and treatment of EC. 展开更多
关键词 poly(A)-specific ribonuclease Esophageal cancer APOPTOTIC Phosphatidylinositol 3-kinase/protein kinase B
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Dietary saturated fatty acid and polyunsaturated fatty acid oppositely affect hepatic NOD-like receptor protein 3 inflammasome through regulating nuclear factor-kappa B activation 被引量:11
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作者 Yong-heng Sui Wen-jing Luo +1 位作者 Qin-Yu Xu jing hua 《World Journal of Gastroenterology》 SCIE CAS 2016年第8期2533-2544,共12页
AIM: To investigate the effect of different dietary fatty acids on hepatic inflammasome activation.METHODS: Wild-type C57BL/6 mice were fed either a high-fat diet or polyunsaturated fatty acid(PUFA)-enriched diet. Pri... AIM: To investigate the effect of different dietary fatty acids on hepatic inflammasome activation.METHODS: Wild-type C57BL/6 mice were fed either a high-fat diet or polyunsaturated fatty acid(PUFA)-enriched diet. Primary hepatocytes were treated with either saturated fatty acids(SFAs) or PUFAs as well as combined with lipopolysaccharide(LPS). The expression of NOD-like receptor protein 3(NLRP3) inflammasome, peroxisome proliferator-activated receptor-γ and nuclear factor-kappa B(NF-κB) was determined by real-time PCR and Western blot. The activity of Caspase-1 and interleukine-1β production were measured.RESULTS: high-fat diet-induced hepatic steatosis was sufficient to induce and activate hepatic NLRP3 inflammasome. SFA palmitic acid(PA) directly activated NLRP3 inflammasome and increased sensitization to LPS-induced inflammasome activation in hepatocytes. In contrast, PUFA docosahexaenoic acid(Dh A) had thepotential to inhibit NLRP3 inflammasome expression in hepatocytes and partly abolished LPS-induced NLRP3 inflammasome activation. Furthermore, a highfat diet increased but PUFA-enriched diet decreased sensitization to LPS-induced hepatic NLRP3 inflammasome activation in vivo. Moreover, PA increased but Dh A decreased phosphorylated NF-κB p65 protein expression in hepatocytes.CONCLUSION:Hepatic NLRP 3 inflammasome activation played an important role in the development of non-alcoholic fatty liver disease. Dietary SFAs and PUFAs oppositely regulated the activity of NLRP3 inflammasome through direct activation or inhibition of NF-κB. 展开更多
关键词 Non-alcoholic FATTY liver disease NODlike receptor protein 3 INFLAMMASOME Saturated FATTY ACIDS poly
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Interaction of Immune System Protein with PEGylated and Un-PEGylated Polymeric Nanoparticles
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作者 Matthewos Eshete Kayla Bailey +2 位作者 Tuyen Duong Thanh Nguyen Santosh Aryal Seong-O Choi 《Advances in Nanoparticles》 2017年第3期103-113,共11页
Biodegradable Nanoparticles (NPs) are under intense investigation due to their potential application in targeted drug delivery. Upon their entry to the biological system, they encounter the immune system, which limits... Biodegradable Nanoparticles (NPs) are under intense investigation due to their potential application in targeted drug delivery. Upon their entry to the biological system, they encounter the immune system, which limits their availability at the intended site. Most importantly, the innate immune system is the one that acts as the first line of defense against foreign materials. It can be activated by collectin proteins which recognize the structural pattern of polysaccharide on the surface of microorganisms. NPs may interact with these proteins in a similar way, and the interaction may lead to beneficial outcomes in vaccine delivery. On the other hand, in targeted drug delivery, it is desirable for the NPs not to be recognized as foreign material as this may lead to their fast elimination from the system through mechanism such as opsonization. We investigated the interaction of PEGylated and un-PEGylated PLGA NPs with Recombinant Human Mannose-Binding Protein (HMBP) in an effort to understand the effect of surface modification on their binding to the protein. Results show that both PLGA-COOH and PLGA-PEG-NH2 bind to HMBP as studied using dynamic light scattering (DLS), fluoresce and UV-vis spectroscopy. However, their binding is shown to have different effect on the structure of the protein. Study done using fluorescence spectroscopy displayed a decrease in fluorescence emission of the protein upon binding to PLGA-COOH. On the other hand the fluorescence emission of the protein increased upon binding to the PLGA-PEG-NH2 indicating conformational changes in the protein structure. 展开更多
关键词 Nanoparticles Mannose-binding protein poly (L-Lactic-Co-Glycolic Acid) NANOPARTICLE protein INTERACTION
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Compatibility of Soybean Protein and Poly (vinyl alcohol)
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作者 沈婕 肖茹 顾利霞 《Journal of Donghua University(English Edition)》 EI CAS 2004年第4期26-28,共3页
Bicomponent fibers were wet-spun from soybean protein and poly (vinyl alcohol). The fiber was brittle and showed a high frequency of breakage upon drawing and the bad compatibility between soybean protein and poly (vi... Bicomponent fibers were wet-spun from soybean protein and poly (vinyl alcohol). The fiber was brittle and showed a high frequency of breakage upon drawing and the bad compatibility between soybean protein and poly (vinyl alcohol) was thought to be the causes for the poor drawability. Our effort was then to study the soybean protein and poly (vinyl alcohol) solution, with the aim of trying to improve the components’ compatibility and to determine the proper solution condition for dissolving them. The effects of alkali, sodium sulfite and urea on the compatibility of the solution were examined. 展开更多
关键词 兼容性 大豆蛋白质 聚乙烯醇 双组分纤维 湿纺
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Ribotrap Analysis of Proteins Associated with FHL3 3'Untranslated Region in Glioma Cells
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作者 Wei Han Qing Xia +1 位作者 Bin Yin Xiao-zhong Peng 《Chinese Medical Sciences Journal》 CAS CSCD 2014年第2期78-84,共7页
Objective To screen the proteins associated with four-and-a-half LIM domains 3(FHL3) 3' untranslated region(3'UTR) in glioma cells. Methods Western blot was adopted to detect the regulatory effect of poly(C)-b... Objective To screen the proteins associated with four-and-a-half LIM domains 3(FHL3) 3' untranslated region(3'UTR) in glioma cells. Methods Western blot was adopted to detect the regulatory effect of poly(C)-binding protein 2(PCBP2) on FHL3. Biotin pull-down and sliver staining were employed to screen and verify the candidate binding proteins of FHL3 3'UTR. Then liquid chromatography-tandem mass spectrometry(LC-MS/MS) and molecule annotation system were used to identify and analyze the candidate binding proteins. Immunoprecipitation was conducted to study the interaction between PCBP2 and polypyrimidine tract-binding protein 1(PTBP1), a binding protein identified by LC-MS/MS. Results PCBP2 could bind to FHL3 mRNA 3'UTR-A and inhibited the expression of FHL3 in T98 G glioms cells. 22 candidate binding proteins were identified. Among them, there were 11 RNA binding proteins, including PCBP2. PTBP1 associated with FHL3 mRNA 3'UTR and interacted with PCBP2 protein. Conclusion PCBP2 and PTBP1 can both associate with FHL3 mRNA 3'UTR through forming a protein complex. 展开更多
关键词 胶质瘤细胞 非翻译区 蛋白质 RNA结合蛋白 LC-MS LIM结构域 串联质谱 mRNA
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Lnc-PCIR通过调控PABPC4的泛素化促进喉癌细胞的增殖和迁移
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作者 张杨 邓欣欣 +3 位作者 王珍 杨丽萍 张莹莹 梁耕田 《现代肿瘤医学》 CAS 2024年第7期1228-1235,共8页
目的:探究LncRNA RP11-214F16.8(Lnc-PCIR)通过调控多腺苷酸结合蛋白质4[poly(A)-binding protein cytoplasmic 4,PABPC4]的泛素化对喉癌细胞增殖、迁移的影响。方法:利用Human Protein Atlas、GEPIA数据库分析Lnc-PCIR、PABPC4在头颈... 目的:探究LncRNA RP11-214F16.8(Lnc-PCIR)通过调控多腺苷酸结合蛋白质4[poly(A)-binding protein cytoplasmic 4,PABPC4]的泛素化对喉癌细胞增殖、迁移的影响。方法:利用Human Protein Atlas、GEPIA数据库分析Lnc-PCIR、PABPC4在头颈鳞状细胞癌组织中的水平及患者的总体生存期。选取我院130例喉癌患者的癌及癌旁组织标本,RT-qPCR、Western blot检测组织Lnc-PCIR、PABPC4 mRNA和蛋白水平,并分析Lnc-PCIR水平与患者临床病理参数的关系。将HEP-2、TU212细胞进行不同转染,分为si-Lnc-NC组、si-Lnc-PCIR组、Lnc-NC组、Lnc-PCIR组、si-Lnc-PCIR+NC组、si-Lnc-PCIR+PABPC4组。CCK-8实验、EdU染色和Transwell实验检测细胞增殖和迁移能力;RT-qPCR检测细胞Lnc-PCIR、PABPC4 mRNA水平;HEP-2、TU212细胞分别经放线菌酮(cycloheximide,CHX)、MG132、ML364处理后,Western blot检测细胞PABPC4蛋白水平。20只裸鼠经皮下注射已转染的HEP-2细胞悬液,分为si-Lnc-NC组、si-Lnc-PCIR组;1个月后,测定移植瘤体积、质量和Lnc-PCIR、PABPC4蛋白水平。结果:数据库显示头颈鳞状细胞癌组织中Lnc-PCIR、PABPC4水平高于正常组织,且Lnc-PCIR、PABPC4水平与总体生存期呈负相关(均P<0.05)。与正常组织相比,患者喉癌组织中Lnc-PCIR、PABPC4 mRNA和蛋白水平升高,且Lnc-PCIR水平与患者性别、TNM分期、肿瘤分化程度、淋巴结转移相关(均P<0.05)。敲低Lnc-PCIR后,细胞Lnc-PCIR、PABPC4蛋白水平降低;细胞培养1 d、2 d、3 d后的OD值、EdU阳性率降低,细胞迁移数减少。而过表达PABPC4能部分逆转敲低Lnc-PCIR对细胞增殖和迁移的影响(均P<0.05);过表达Lnc-PCIR能降低细胞PABPC4泛素化水平(P<0.05)。敲低Lnc-PCIR能抑制小鼠移植瘤生长(P<0.05)。结论:Lnc-PCIR通过调控PABPC4的泛素化促进喉癌细胞的增殖和迁移。 展开更多
关键词 Lnc-PCIR 多腺苷酸结合蛋白质4 泛素化 喉癌
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木薯花叶病毒AC4蛋白与AtPARN互作研究 被引量:1
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作者 刘琳玉 赵平娟 +3 位作者 符艳 刘志昕 任艳利 张秀春 《热带作物学报》 CSCD 北大核心 2024年第1期197-204,共8页
木薯(Manihot esculenta Crantz)作为重要的热带作物,是全球六大粮食作物之一,并为全球近七亿人口提供主粮,然而病毒侵染引起的病害在全球范围内严重威胁木薯产业的发展。木薯花叶病毒病(cassava mosaic disease,CMD)是最有威胁的病害之... 木薯(Manihot esculenta Crantz)作为重要的热带作物,是全球六大粮食作物之一,并为全球近七亿人口提供主粮,然而病毒侵染引起的病害在全球范围内严重威胁木薯产业的发展。木薯花叶病毒病(cassava mosaic disease,CMD)是最有威胁的病害之一,造成严重的经济损失。斯里兰卡木薯花叶病毒(SriLankancassavamosaicvirus,SLCMV)是引发木薯花叶病的病原物之一,SLCMV是典型的双组分双生病毒,其基因组由DNA-A和DNA-B两个环状组分组成。无义介导的mRNA降解(nonsense-mediatedmRNAdecay,NMD)是真核细胞mRNA降解的主要途径之一,也是真核生物普遍存在的抗病毒防御机制。越来越多的研究显示,NMD不仅是真核生物重要的mRNA数量、质量调控机制,还与转录后基因沉默(post transcriptional gene silencing,PTGS)同样能降解病毒RNA,是真核生物普遍存在的抗病毒防御机制。NMD的mRNA衰减过程包括PTC的识别、脱腺苷酸、脱帽和最后的核酸外切酶降解4个过程,UPF1、PARN、DCP2和XRN4分别是上述4个过程中的关键蛋白。病毒是专性寄生生物,必须逃避或耐受寄主细胞的降解,才能成功感染。聚腺苷酸特异性核糖核酸酶[poly(A)-specific ribonuclease,PARN]是NMD信号通路的一个关键因子。目前,关于SLCMV抵御寄主NMD的分子机制尚不明确。本研究采用酵母双杂交(yeast two-hybrid system)和荧光双分子互补(bimolecular fluorescence complementation,BiFC)试验证明斯里兰卡木薯花叶病毒(SLCMV)编码的沉默抑制子AC4与拟南芥PARN相互作用。据此推测SLCMV AC4蛋白可能通过与AtPARN相互作用而抑制寄主NMD的病毒防御功能帮助病毒逃避或耐受寄主细胞的降解。研究结果为阐明木薯花叶病毒调控寄主NMD抗病毒防御功能的分子机理奠定基础。 展开更多
关键词 斯里兰卡木薯花叶病毒(SLCMV) AC4蛋白 聚腺苷酸特异性核糖核酸酶(PARN) 酵母双杂交(Y2H) 荧光双分子互补(BiFC)
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γ-聚谷氨酸和鱼蛋白水溶肥对松花菜生长和产量的影响 被引量:1
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作者 李方勇 刘庆叶 +9 位作者 黄忠阳 吴旭东 戎茸 张宗俊 陈莉莉 胡卫丛 王蓓 徐永安 王东升 李伟明 《长江蔬菜》 2024年第4期68-71,共4页
为明确γ-聚谷氨酸和鱼蛋白水溶肥在蔬菜生产上的作用,以松花菜为供试作物,设置常规施肥、γ-聚谷氨酸、鱼蛋白3个处理。试验结果表明,γ-聚谷氨酸处理和鱼蛋白处理都能显著提高松花菜的株高、茎粗,提升松花菜品质和产量,增加松花菜的... 为明确γ-聚谷氨酸和鱼蛋白水溶肥在蔬菜生产上的作用,以松花菜为供试作物,设置常规施肥、γ-聚谷氨酸、鱼蛋白3个处理。试验结果表明,γ-聚谷氨酸处理和鱼蛋白处理都能显著提高松花菜的株高、茎粗,提升松花菜品质和产量,增加松花菜的经济效益,以鱼蛋白处理对松花菜增产增效效果更好。 展开更多
关键词 新型肥料 Γ-聚谷氨酸 鱼蛋白
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小白链霉菌全细胞转化L-赖氨酸合成ε-聚赖氨酸的体系构建与优化
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作者 朱道君 刁文娇 +4 位作者 张佳微 王靓 张宏建 张建华 陈旭升 《食品与发酵工业》 CSCD 北大核心 2024年第1期29-36,共8页
小白链霉菌(Streptomyces albulus)是天然抗菌肽ε-聚赖氨酸(ε-poly-L-lysine,ε-PL)的主要生产菌株。为了提高小白链霉菌生产ε-PL效率,该文构建并优化了全细胞转化L-赖氨酸合成ε-PL体系:葡萄糖质量浓度80 g/L,菌龄12 h,反应温度30℃... 小白链霉菌(Streptomyces albulus)是天然抗菌肽ε-聚赖氨酸(ε-poly-L-lysine,ε-PL)的主要生产菌株。为了提高小白链霉菌生产ε-PL效率,该文构建并优化了全细胞转化L-赖氨酸合成ε-PL体系:葡萄糖质量浓度80 g/L,菌龄12 h,反应温度30℃,L-赖氨酸质量浓度15 g/L,柠檬酸浓度15 g/L,初始反应p H 4.0,硫酸铵质量浓度6 g/L,湿菌体量为1900 g/L。基于该转化体系,实现小白链霉菌在96 h合成ε-PL产量和底物转化率达到13.80 g/L和38.9%,分别是常规摇瓶发酵的4.1、3.2倍。最后,在小白链霉菌中异源表达来自大肠杆菌的L-赖氨酸特异性通透蛋白基因lysp,获得的重组菌S.albulus OE-lysp实现L-赖氨酸利用能力和底物转化率较出发菌株分别提升26%和33%,ε-PL产量增加至17.21 g/L,约为常规摇瓶发酵ε-PL产量的6.4倍,这是文献报道的最高摇瓶规模ε-PL产量。该研究结果一方面说明了通过全细胞转化L-赖氨酸生产ε-PL的可行性,另一方面为S.albulus转化大宗氨基酸L-赖氨酸生产高值ε-PL奠定了坚实的技术基础,具有重要的理论意义和经济价值。 展开更多
关键词 小白链霉菌 Ε-聚赖氨酸 全细胞转化 异源表达 L-赖氨酸通透蛋白
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Poly I:C母鼠免疫刺激及母婴分离对子代精神分裂症样行为的影响 被引量:5
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作者 王玉聪 王佳 +10 位作者 李国海 冯驰 姜扬 陈嘉欣 王文欣 彭灏 侯卓然 徐家丹 肖栩 汪磊 张维宁 《江苏大学学报(医学版)》 CAS 2017年第5期379-384,390,共7页
目的:探讨聚肌苷酸胞苷酸(polyinosinic-polycytidylic acid,poly I:C)诱导的妊娠母体免疫刺激和出生后母婴分离双重因素对子代精神分裂症样行为的影响及其机制。方法:于母鼠妊娠第17天时尾静脉注射poly I:C(5 mg/kg)或无菌生理盐水,子... 目的:探讨聚肌苷酸胞苷酸(polyinosinic-polycytidylic acid,poly I:C)诱导的妊娠母体免疫刺激和出生后母婴分离双重因素对子代精神分裂症样行为的影响及其机制。方法:于母鼠妊娠第17天时尾静脉注射poly I:C(5 mg/kg)或无菌生理盐水,子代小鼠出生后每窝随机选取一半进行母婴分离,另一半不分离。子代鼠出生第21天断奶分笼,第60天进行行为学实验:开场实验、高架十字迷宫实验、新物体/位置识别实验以及水迷宫实验。行为实验结束后,处死小鼠取海马组织,采用蛋白质印迹法检测髓鞘碱性蛋白(myelin basic protein,MBP)的表达。结果:高架十字迷宫实验结果显示,poly I:C可导致雄性小鼠出现抗焦虑;新物体识别实验显示,poly I:C可影响雌子代的认知功能;水迷宫实验结果显示poly I:C对雌、雄仔鼠的学习记忆能力有不同程度的影响,雌性仔鼠颠倒学习能力受损;此外,poly I:C导致雌、雄仔鼠海马MBP降低。母婴分离导致雌、雄仔鼠的体重增长速度减缓、雌性后代的颠倒学习能力降低。结论:poly I:C诱导的母体免疫刺激与母婴分离,均能诱导子代出现精神分裂症样行为表现,且对雌、雄子代的影响不尽相同,两种干预因素没有出现协同或拮抗作用;此外,poly I:C导致了子代的海马髓鞘损伤。 展开更多
关键词 精神分裂症动物模型 poly I:C 妊娠 母婴分离 髓鞘碱性蛋白
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小麦返白系返白期间Poly(A)RNA与蛋白质水平的变化 被引量:4
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作者 徐恒平 汪沛洪 《植物生理学报(0257-4829)》 CSCD 1992年第4期337-340,共4页
比较了小麦返白系与其原始品种矮变1号在返白期间Poly(A)RNA与蛋白质水平的变化。在心叶变白过程中。返白系的蛋白质含量下降,随着复绿又回升;而RNA含量变化与此大致呈平行关系。在全白阶段,返白系的Poly(A)RNA含量大幅度下降。体外合... 比较了小麦返白系与其原始品种矮变1号在返白期间Poly(A)RNA与蛋白质水平的变化。在心叶变白过程中。返白系的蛋白质含量下降,随着复绿又回升;而RNA含量变化与此大致呈平行关系。在全白阶段,返白系的Poly(A)RNA含量大幅度下降。体外合成的蛋白质也少得多,但其翻译活性/Poly(A)RNA及各期的蛋白质/Poly(A)RNA比例并不比对照低。可能返白系在返白期间蛋白质水平的变化与转录水平的基因调控有关。 展开更多
关键词 小麦 多聚核糖核酸 蛋白
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家蚕Poly(A)结合蛋白基因BmPABP的克隆及序列与功能分析
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作者 徐汉福 王峰 +3 位作者 王根洪 段晓利 马三垣 夏庆友 《蚕业科学》 CAS CSCD 北大核心 2011年第3期412-418,共7页
Poly(A)结合蛋白[Poly(A)-binding protein,PABP]是广泛存在于真核生物细胞内的一类高保守性蛋白,通过与Poly(A)的结合形成翻译起始复合物调节mRNA的稳定性和翻译效率。利用RT-PCR方法克隆了家蚕Poly(A)结合蛋白基因Bm-PABP,该基因含有... Poly(A)结合蛋白[Poly(A)-binding protein,PABP]是广泛存在于真核生物细胞内的一类高保守性蛋白,通过与Poly(A)的结合形成翻译起始复合物调节mRNA的稳定性和翻译效率。利用RT-PCR方法克隆了家蚕Poly(A)结合蛋白基因Bm-PABP,该基因含有4个外显子,开放阅读框(open reading frame,ORF)长度为1 812 bp,编码603个氨基酸,具有4个典型的RNA识别模体(RNA recognization motif,RRM)和1个保守的C末端结构域。基因芯片数据分析显示,BmPABP在家蚕5龄第3天幼虫各组织以及5龄第4天至化蛾阶段具有高水平的转录表达。构建基于家蚕Actin4启动子的瞬时转染载体pSLA4-Bm-PABP,将其与携带有荧光素酶报告基因的pSLA4-LUC质粒分别按1∶1和2∶1的摩尔比混合后共转染昆虫Sf9细胞,于转染后72 h检测荧光素酶活性,结果显示荧光素酶的活性较对照分别提高了11.9倍和7.5倍。研究结果初步证实了BmPABP具有促进目的基因表达的功能,同时提示利用BmPABP有望提高外源基因在转基因家蚕中的表达水平,可应用于家蚕生物反应器研究。 展开更多
关键词 家蚕 poly(A)结合蛋白 基因克隆 序列结构 细胞转染 功能
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Lipid-Based Poly(I:C) Adjuvants Strongly Enhance the Immunogenicity of SARS-CoV-2 Receptor-Binding Domain Vaccine
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作者 Yixin Wu Liuxian Meng +3 位作者 Huicong Zhang Shun Hu Fusheng Li Yingjie Yu 《Infectious Diseases & Immunity》 CSCD 2023年第1期3-12,共10页
Background The outbreak of the severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)has greatly threatened public health.Recent studies have revealed that the spike receptor-binding domain(RBD)of SARS-CoV-2 is a... Background The outbreak of the severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)has greatly threatened public health.Recent studies have revealed that the spike receptor-binding domain(RBD)of SARS-CoV-2 is a potent target for vaccine development.However,adjuvants are usually required to strengthen the immunogenicity of recombinant antigens.Different types of adjuvants can elicit different immune responses.Methods We developed an RBD recombinant protein vaccine with a polyriboinosinic acid–polyribocytidylic acid[poly(I:C)]adjuvant to evoke a strong immune response.The delivery of poly(I:C)was optimized in two steps.First,poly(I:C)was complexed with a cationic polymer,poly-l-lysine(PLL),to form poly(I:C)–PLL,a polyplex core.Thereafter,it was loaded into five different lipid shells(group II,III-1,2-distearoyl-sn-glycero-3-phosphocholine[DSPC],III-1,2-dioleoyl-sn-glycero-3-phosphoethanolamine[DOPE],IV-DOPE,and IV-DSPC).We performed an enzyme-linked immunosorbent assay and enzyme-linked immunosorbent spot assay to compare the ability of the five lipopolyplex adjuvants to enhance the immunogenicity of the SARS-CoV-2 RBD protein,including humoral and cellular immune responses.Finally,the adjuvant with the highest immunogenicity was selected to verify the protective immunity of the vaccine through animal challenge experiments.Results Recombinant RBD protein has low immunogenicity.The different adjuvants we developed enhanced the immunogenicity of the RBD protein in different ways.Among the lipopolyplexes,those containing DOPE(III-DOPE and IV-DOPE)elicited RBD-specific immunoglobulin G antibody responses,and adjuvants with four components elicited better RBD-specific immunoglobulin G antibody responses than those containing three components(P<0.05).The IC50 and IC90 titers indicated that the IV-DOPE lipopolyplex had the greatest neutralization ability,with IC50 titers of 1/117,490.Furthermore,in the challenge study,IV-DOPE lipopolyplex protected mice from SARS-CoV-2 infection.On the fourth day after infection,the average animal body weights were reduced by 18.56%(24.164±0.665 g vs.19.678±0.455 g)and 0.06%(24.249±0.683 g vs.24.235±0.681 g)in the MOCK and vaccine groups,respectively.In addition,the relative expression of viral RNA in the vaccinated group was significantly lower than that in the MOCK group(P<0.05).Interstitial inflammatory cell infiltration was observed in the MOCK group,whereas no obvious damage was observed in the vaccinated group.Conclusions The IV-DOPE–adjuvanted SARS-CoV-2 recombinant RBD protein vaccine efficiently protected mice from SARS-CoV-2 in the animal challenge study.Therefore,IV-DOPE is considered an exceptional adjuvant for SARS-CoV-2 recombinant RBD protein-based vaccines and has the potential to be further developed into a SARS-CoV-2 recombinant RBD protein-based vaccine. 展开更多
关键词 Adjuvants vaccine Lipopolyplex poly(I:C) RBD protein
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Design,synthesis and characterization of novel biodegradable macrodiols based on poly(DL-lactic acid) and poly(p-dioxanone) 被引量:2
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作者 Chan Yu Zhang Yan Feng Luo +3 位作者 Su Jun Wang Zhao Liu Yuan Liang Wang Zhi Qing Liang 《Chinese Chemical Letters》 SCIE CAS CSCD 2009年第6期743-746,共4页
Integrating poly(lactic acid) (PLA), glycolic acid (GA) and ethylene glycol (EG) will hopefully result in a novel copolymer that combines such advantages as fastened and controllable release rate and improved ... Integrating poly(lactic acid) (PLA), glycolic acid (GA) and ethylene glycol (EG) will hopefully result in a novel copolymer that combines such advantages as fastened and controllable release rate and improved flexibility together with good biocompatibility. In this study, p-dioxanone (PDO) was employed to copolymerize with DL-lactide (LA) via ring-opening melt polymerization using Sn(Oct)2 as an initiator and ethylene glycol as a co-initiator. The obtained degradable macrodiols (HO-P(LA-co-PDO)-OH) were just such a copolymer consisting of PLA, GA and EG. 1HNMR was employed to characterize the copolymers, and the effect of PDO/LA molar ratios in the feedstock on the molecular weights of HO-P(LA-co-PDO)-OH was investigated by means of endhydroxyl analysis, 1H NMR or GPC-MALLs. The results confirmed the successful synthesis of HO-P(LA-co-PDO)-OH and revealed that one end-hydroxyl of the micarodiols was donated by LA or PDO and the other one by the co-initiator EG. In addition, the molecular weights of HO-P(LA-co-PDO)-OH increased with decreasing PDO/LA ratios. 展开更多
关键词 Macrodiol poly(lactici acid) P-DIOXANONE Ethylene glycol Drug delivery carder Bone morphogenetic protein
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A Temperature-sensitive Hydrogel Refolding System: Preparation of Poly(N-isopropyl acrylamide) and Its Application in Lysozyme Refolding 被引量:6
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作者 崔志芳 关怡新 姚善泾 《Chinese Journal of Chemical Engineering》 SCIE EI CAS CSCD 2004年第4期556-560,共5页
Temperature-sensitive hydrogel—poly(N-isopropyl acrylamide) (PNIPA) was prepared and applied to protein refolding. PNIPA gel disks and gel particles were synthesized by the solution polymerization and inverse suspens... Temperature-sensitive hydrogel—poly(N-isopropyl acrylamide) (PNIPA) was prepared and applied to protein refolding. PNIPA gel disks and gel particles were synthesized by the solution polymerization and inverse suspension polymerization respectively. The swelling kinetics of the gels was also studied. With these prepared PNIPA gels, the model protein lysozyme was renatured. Within 24h, PNIPA gel disks improved the yield of lysozyme activity by 49.3% from 3375.2U·mg^-1 to 5038.8U·mg^-1. With the addition of faster response PNIPA gel beads, the total lysozyme activity recovery was about 68.98% in 3h, as compared with 42.03% by simple batch dilution. The novel refolding system with PNIPA enables efficient refolding especially at high protein concentrations. Discussion about the mechanism revealed that when PNIPA gels were added into the refolding buffer, the hydrophobic interactions between denatured proteins and polymer gels could prevent the aggregation of refolding intermediates, thus enhanced the protein renaturation. 展开更多
关键词 温度 敏感性 水凝胶 聚乙烯 DNA 蛋白质 异丙基丙烯酰胺
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POLY(ETHERSULFONE)/N,O-CARBOXYMETHYL AMPHOTERIC CHITOSAN COMPOSITE MF MEMBRANE: PREPARATION AND ITS CHARGED CHARACTERISTICS 被引量:6
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作者 赵之平 王志 +1 位作者 叶楠 王世昌 《化工学报》 EI CAS CSCD 北大核心 2002年第1期2-3,共2页
关键词 聚醚砜 N O-羧甲基两性壳聚糖 复合微滤膜 膜材料 膜分离 制备 荷电特性
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APPLICATION OF NONIONIC TEMPERATURE SENSITIVE HYDROGEL FOR CONCENTRATION OF PROTEIN AQUEOUS SOLUTION
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作者 孙以实 邱志勇 洪耀良 《Chinese Journal of Polymer Science》 SCIE CAS CSCD 1992年第4期311-318,共8页
Six different N-alkyl substituted acrylarnide nonionic hydrogels were prepared and their swelling characteristics were measured. Poly N-isopropyl acrylamide (PNIPA) and poly N-n-propyl-acrylamide (PNNPA) temperature s... Six different N-alkyl substituted acrylarnide nonionic hydrogels were prepared and their swelling characteristics were measured. Poly N-isopropyl acrylamide (PNIPA) and poly N-n-propyl-acrylamide (PNNPA) temperature sensitive hydrogels were chosen as the nonionic temperature sensitive hydrogels for concentration of very dilute aqueous protein solution. The separation properties of PNIPA and PNNPA hydr0gels with different network dimensions were studied and the modification of the hydrogels was surveyed in order to decrease their surface adsorption of protein molecules. The experimental results of the concentration of BSA (Bovin serum albumin) dilute aqueous solution by hydroxylpropyl methacrylate (HPMA) copolymerized PNIPA hydrogel were given. The value and the limitation of concentration of dilute aqueous protein solution by this method was evaluated. 展开更多
关键词 poly N-n-propyl acrylamide poly N-isopropyl acrlamide Hydroxyl propyl methacrylate thermally sensitive hydrogel Concentration of protein solution Bovin serum albumin.
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Polyurethane/poly(vinyl alcohol) hydrogel coating improves the cytocompatibility of neural electrodes
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作者 Mei Li Hai-han Zhou +3 位作者 Tao Li Cheng-yan Li Zhong-yuan Xia Yanwen Y.Duan 《Neural Regeneration Research》 SCIE CAS CSCD 2015年第12期2048-2053,共6页
Neural electrodes,the core component of neural prostheses,are usually encapsulated in polydimethylsiloxane(PDMS).However,PDMS can generate a tissue response after implantation.Based on the physicochemical properties... Neural electrodes,the core component of neural prostheses,are usually encapsulated in polydimethylsiloxane(PDMS).However,PDMS can generate a tissue response after implantation.Based on the physicochemical properties and excellent biocompatibility of polyurethane(PU)and poly(vinyl alcohol)(PVA)when used as coating materials,we synthesized PU/PVA hydrogel coatings and coated the surface of PDMS using plasma treatment,and the cytocompatibility to rat pheochromocytoma(PC12)cells was assessed.Protein adsorption tests indicated that the amount of protein adsorption onto the PDMS substrate was reduced by 92%after coating with the hydrogel.Moreover,the PC12 cells on the PU/PVA-coated PDMS showed higher cell density and longer and more numerous neurites than those on the uncoated PDMS.These results indicate that the PU/PVA hydrogel is cytocompatible and a promising coating material for neural electrodes to improve their biocompatibility. 展开更多
关键词 nerve regeneration cerebral injury neural electrodes hydrogel coatings polyURETHANE polydimethylsiloxane poly(vinyl alcohol) CYTOCOMPATIBILITY protein adsorption nerve growth factor mt pheochromocytoma cells synaptic differentiation neural regeneration
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Poly Ⅰ:C刺激对裸鼹鼠和小鼠巨噬细胞PKR/eIF2α信号通路影响的比较研究 被引量:1
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作者 林丽芳 张成财 +6 位作者 李煜 张倩倩 陈超 李壘辰 杨文静 李莉 崔淑芳 《实验动物与比较医学》 CAS 2019年第6期473-478,共6页
目的比较研究聚肌胞苷酸(Poly Ⅰ:C)对裸鼹鼠和小鼠巨噬细胞双链RNA依赖性蛋白激酶R(PKR)/真核细胞翻译起始因子2α(EIF2α)信号通路的影响。方法分别提取裸鼹鼠和C57BL/6J小鼠巨噬细胞,随机分成对照组、Poly Ⅰ:C给药组、2-氨基嘌呤(2-... 目的比较研究聚肌胞苷酸(Poly Ⅰ:C)对裸鼹鼠和小鼠巨噬细胞双链RNA依赖性蛋白激酶R(PKR)/真核细胞翻译起始因子2α(EIF2α)信号通路的影响。方法分别提取裸鼹鼠和C57BL/6J小鼠巨噬细胞,随机分成对照组、Poly Ⅰ:C给药组、2-氨基嘌呤(2-AP)给药组、Poly Ⅰ:C+2-AP给药组。给药后,蛋白印迹检测细胞中磷酸化PKR(Pho-PKR)、PKR、磷酸化EIF2α(Pho-EIF2α)、EIF2α、Caspase-8蛋白的表达情况。结果 Poly Ⅰ:C给药后,小鼠巨噬细胞的PKR和e IF2α磷酸化水平显著降低(P<0.01),而裸鼹鼠相反则显著升高(P<0.01),表明Poly Ⅰ:C能抑制小鼠的PKR活性,而裸鼹鼠的PKR活性被显著激活;Poly Ⅰ:C干预的基础上通过2-AP抑制PKR活性,结果显示,2-AP给药后下调了小鼠和裸鼹鼠PKR和磷酸化PKR的表达(P<0.01),尤其是裸鼹鼠的PKR,2-AP给药后显著地抑制裸鼹鼠PKR的表达;单独2-AP给药后小鼠巨噬细胞的活性PKR和活性eIF2α比例显著下降(P<0.01),裸鼹鼠巨噬细胞的活性PKR比例也显著下降(P<0.01);通过2-AP抑制PKR活性后,裸鼹鼠细胞的Caspase-8蛋白表达水平显著下降,但小鼠细胞的表达却无显著差异。结论与小鼠比较,裸鼹鼠细胞对Poly Ⅰ:C刺激具有较高的抗性;细胞的PKR对2-AP的敏感性更强;PKR活性对Caspase-8表达有促进作用。 展开更多
关键词 裸鼹鼠 聚肌胞苷酸(polyⅠ:C) 双链RNA依赖性蛋白激酶R(PKR) CASPASE-8
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二甲双胍改善由C9ORF72肌萎缩侧索硬化/额颞叶痴呆相关多聚甘氨酸-精氨酸诱导的线粒体损伤
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作者 冯奕源 徐忠匀 +2 位作者 尹雅芙 王辉 程维维 《上海交通大学学报(医学版)》 CAS CSCD 北大核心 2023年第7期839-847,共9页
目的·探索C9ORF72肌萎缩侧索硬化/额颞叶痴呆(amyotrophic lateral sclerosis/frontotemporal dementia,ALS/FTD)相关的多聚甘氨酸-精氨酸(poly-glycine-arginine,poly-GR)对线粒体形态及功能的影响,并分析二甲双胍对由poly-GR诱... 目的·探索C9ORF72肌萎缩侧索硬化/额颞叶痴呆(amyotrophic lateral sclerosis/frontotemporal dementia,ALS/FTD)相关的多聚甘氨酸-精氨酸(poly-glycine-arginine,poly-GR)对线粒体形态及功能的影响,并分析二甲双胍对由poly-GR诱导的线粒体损伤的修复作用及其可能的机制。方法·采用慢病毒感染法分别构建能够稳定表达50个重复甘氨酸-精氨酸序列[(glycine-arginine)50,(GR)_(50)]和绿色荧光蛋白(green fluorescent protein,GFP)的SK-N-SH细胞,即(GR)_(50)-SK细胞株和GFP CTRL-SK细胞株。采用蛋白质印迹法(Western blotting)验证已构建细胞中(GR)_(50)蛋白的表达水平,并采用荧光显微镜观察GFP CTRL-SK细胞的GFP表达。采用碘化丙啶(propidium iodine,PI)染色分别检测(GR)_(50)-SK、GFP CTRL-SK细胞的凋亡水平。利用免疫荧光(immunofluorescence,IF)染色分析(GR)_(50)蛋白在细胞内的定位情况。采用超氧化物指示剂MitoSOX Red分别对(GR)_(50)-SK、GFP CTRL-SK细胞的氧自由基进行染色,并利用荧光显微镜观察红色荧光强度以评估线粒体活性氧水平的变化。采用透射电镜分别观察(GR)_(50)-SK、GFP CTRL-SK细胞的线粒体形态。采用Western blotting检测(GR)_(50)-SK、GFP CTRL-SK细胞的蛋白激酶B(protein kinase B,PKB,又称AKT)及其磷酸化水平。利用SC79激活(GR)_(50)-SK细胞中的AKT,并采用MitoSOX Red染色及PI染色实验分析AKT磷酸化后细胞的线粒体活性氧水平及凋亡水平。利用二甲双胍处理(GR)_(50)-SK、GFP CTRL-SK细胞,随后通过上述方法以及ATP检测试剂盒检测细胞的凋亡水平、线粒体活性氧水平、线粒体形态、AKT及其磷酸化水平、ATP浓度情况。结果·Western blotting结果提示(GR)_(50)-SK细胞构建成功,荧光显微镜的观察结果显示GFP CTRL-SK细胞构建成功。PI染色结果显示,(GR)_(50)-SK细胞较GFP CTRL-SK细胞的凋亡水平更高(P=0.016)。IF结果提示,(GR)_(50)-SK细胞中(GR)_(50)蛋白与线粒体存在部分共定位。与GFP CTRL-SK细胞相比,(GR)_(50)-SK细胞的线粒体形态及结构存在明显异常,其活性氧水平明显上升。(GR)_(50)-SK细胞中的AKT水平与GFP CTRL-SK细胞相仿,但磷酸化AKT水平显著下降。SC79处理(GR)_(50)-SK细胞后,可显著上调其AKT磷酸化水平,并下调其活性氧水平及凋亡水平。二甲双胍处理可明显上调(GR)_(50)-SK细胞中的磷酸化AKT水平,但对AKT水平无影响;可重塑该细胞中的部分线粒体形态结构、降低活性氧水平、增加ATP的生成(P=0.000),并下调细胞的凋亡水平(P=0.000)。结论·(GR)_(50)可通过下调AKT磷酸化引起线粒体形态及功能异常,并促进细胞凋亡;而二甲双胍则可抑制由(GR)_(50)蛋白诱导的上述病理事件的发生。 展开更多
关键词 C9ORF72肌萎缩侧索硬化症/额颞叶痴呆 多聚甘氨酸-精氨酸 线粒体 磷酸化蛋白激酶B 二甲双胍
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