目的探讨干扰素诱导的具有四肽重复序列3(interferon induced protein with tetratricopeptide repeats 3,IFIT3)在鼻咽癌CNE-2R细胞中的表达及其对细胞侵袭、转移和上皮-间质转化(epithelial-mesenchymal transition,EMT)能力的影响。...目的探讨干扰素诱导的具有四肽重复序列3(interferon induced protein with tetratricopeptide repeats 3,IFIT3)在鼻咽癌CNE-2R细胞中的表达及其对细胞侵袭、转移和上皮-间质转化(epithelial-mesenchymal transition,EMT)能力的影响。方法采用RT-qPCR和Western blot检测IFIT3在鼻咽癌细胞CNE-2R和正常鼻咽上皮细胞NP69中的表达水平。鼻咽癌CNE-2R细胞分别感染IFIT3 shRNA的3个序列LV1、LV2、LV3,并设立阴性对照组(NC组);利用RT-qPCR和Western blot检测上皮表型E-cadherin及间质表型N-cadherin和Vimentin的表达,采用划痕实验及Transwell实验检测细胞的迁移和侵袭能力。结果IFIT3在鼻咽癌CNE-2R细胞中的mRNA和蛋白表达水平均较NP69细胞显著上调(均P<0.001)。IFIT3在LV1组、LV2组和LV3组中的mRNA和蛋白表达水平均低于NC组(均P<0.001),且以LV2组和LV3组的表达水平最低。与NC组相比,LV2组和LV3组的细胞迁移率显著下降,迁移和侵袭细胞数显著减少,E-cadherin蛋白表达水平显著升高,N-cadherin和Vimentin蛋白表达水平显著降低(均P<0.001)。结论IFIT3在鼻咽癌CNE-2R细胞中表达上调,沉默IFIT3后CNE-2R细胞的侵袭、转移能力以及EMT显著被抑制。展开更多
目的获取小鼠IFIT1(Interferon-induced protein with tetratricopetide repeats 1)特异性多克隆抗体。方法扩增培养高效表达IFIT1的重组子pMAL-C2X-IFIT1,超声破碎后的细胞上清过Amylose Pre-packed column亲和层析柱,将所得纯化的融...目的获取小鼠IFIT1(Interferon-induced protein with tetratricopetide repeats 1)特异性多克隆抗体。方法扩增培养高效表达IFIT1的重组子pMAL-C2X-IFIT1,超声破碎后的细胞上清过Amylose Pre-packed column亲和层析柱,将所得纯化的融合蛋白MBP-IFIT1作为抗原,添加福氏佐机剂后免疫兔,收集抗血清,用Western blot和ELISA方法测定抗血清特异性反应和效价。结果纯化的MBP-IFIT1可诱导兔产生特异性免疫应答,所得抗体能够特异性识别融合蛋白中的IFIT1,ELISA结果显示其效价为1∶12800。结论MBP-IFIT1具有良好的抗原性,以该融合蛋白为抗原免疫兔成功制备出IFIT1特异性多克隆抗体。展开更多
目的检测干扰素诱导蛋白IFIT1(interferon-induced protein with tetratricopetide repeats1)在小鼠各器官组织的表达状况,并在此基础上探讨颈交感神经阻滞(SB)对烧伤后早期小鼠肝脏IFIT1表达的影响。方法雄性C57/129小鼠50只,分为对照...目的检测干扰素诱导蛋白IFIT1(interferon-induced protein with tetratricopetide repeats1)在小鼠各器官组织的表达状况,并在此基础上探讨颈交感神经阻滞(SB)对烧伤后早期小鼠肝脏IFIT1表达的影响。方法雄性C57/129小鼠50只,分为对照组和SB治疗组,TBSA15%～20%Ⅲ度小鼠烧伤模型,分别于烧伤前、烧伤后1、6、12h和24h取肝组织,采用免疫印迹法(Western blot)观察IFIT1的蛋白表达情况,同时提取正常小鼠心、肝、脾、肺、肾、小肠、淋巴细胞和骨骼肌组织RNA,以RT-PCR检测IFIT1表达情况。结果RT-PCR结果显示IFIT1在正常小鼠各组织均有表达,但组织间的表达量存在差异。Western blot结果表明对照组烧伤后肝组织IFIT1表达显著增高(P<0.01),烧伤后SB治疗组IFIT1表达量较对照组相应时相点降低非常显著(P<0.01)。结论IFIT1在小鼠体内广泛表达。SB对严重创伤的治疗作用可能是通过抑制肝内IFIT1表达来实现的。展开更多
目的：观察小鼠心、肝、脾、肺、肾等器官干扰素诱导蛋白IFIT1（Interferon—induced protein with tetratfieopetide repeats1）的表达情况，探讨颈交感神经阻滞（cervical sympathetic block，SB）对烧伤后早期小鼠各器官IFIT1表达状...目的：观察小鼠心、肝、脾、肺、肾等器官干扰素诱导蛋白IFIT1（Interferon—induced protein with tetratfieopetide repeats1）的表达情况，探讨颈交感神经阻滞（cervical sympathetic block，SB）对烧伤后早期小鼠各器官IFIT1表达状况的影响．方法：将雄性昆明小鼠18只随机分为正常对照组、烧伤组和颈交感神经阻滞（SB）治疗组、烧伤组和SB治疗组采用体表总面积（Total body surface area，TBSA）15％-20％Ⅲ度小鼠烧伤模型，SB治疗组于烧伤后即刻进行双侧颈交感神经阻滞，各组分别于4h后取心、肝、脾、肺、肾组织，提取RNA，以RT—PCR检测IFIT1表达情况．结果：RT—PCR显示IFIT1在各组的各器官均有表达，并存在差异．烧伤组IFIT1在心、肝、脾表达低于正常组，同时在肺、肾表达高于正常组，差异显著（P〈0．01）．SB治疗组IFIT1的表达均低于正常组（P〈0．01）；同时，SB治疗组IFIT1在心、肝、肺、肾的表达也低于烧伤组（P〈0．01）．另外，在各组内的不同器官的IFIT1表达量存在差异（P〈0．05）．结论：烧伤后早期小鼠各器官IFIT1表达有不同的变化，可能与不同器官在烧伤应激反应过程中损伤程度不同有关．SB可显著降低各器官IFIT1的表达．展开更多
目的：观察小鼠应激相关蛋白IFIT1（Interferon-induced protein with tetratricopetide repeats1）和JAB1（c-Jun平activation共处五项原则domain-binding protein 1）在肝脏的表达的细胞定位及氯胺酮对烧伤后早期肝脏IFIT1和JAB1表达...目的：观察小鼠应激相关蛋白IFIT1（Interferon-induced protein with tetratricopetide repeats1）和JAB1（c-Jun平activation共处五项原则domain-binding protein 1）在肝脏的表达的细胞定位及氯胺酮对烧伤后早期肝脏IFIT1和JAB1表达的影响，初步探讨氯胺酮调节炎症反应的分子机制。方法：将15只健康C57／129小鼠随机分为3组（n=5），正常对照组、烧伤组、烧伤＋氯胺酮组。采用TBSA 15～20％Ⅲ度小鼠烧伤模型；氯胺酮10mg／kg，肌肉注射，烧伤15min给予。分别于烧伤后4h脱臼处死，取肝。采用免疫印迹法（Western blot）察各组肝IFIT1和JAB1的表达情况。取正常对照组肝组织作病理切片，行免疫组织化学方法对IFIT1和JAB1的表达做细胞定位。结果：烧伤组IFIT1表达较正常对照组显著增高（P〈0．01）．烧伤组JAB1表达较正常对照组显著降低（P〈0．01）：氯胺酮治疗组IFIT1表达较正常对照组、烧伤对照组显著降低（P〈0．01）．氯胺酮治疗组JAB1表达较正常对照组显著降低（P〈0．05）、较烧伤组显著升高（P〈0．05）。免疫组织化学方法显示．IFIT1在肝细胞胞浆表达为阳性．JAB1在肝细胞胞浆和胞核均表达为阳性。结论：烧伤早期小鼠肝组织IFIT1表达增高而JJAB1表达降低，氯胺酮可降低烧伤早期小鼠肝组织IFIT1的高表达，增高JAB1表达；氯胺酮可通过调控严重烧伤应激中IFIT1和JAB1的表达而在全身炎性反应综合症（Systemic inflammatory response syndrome SIRS）中发挥作用。展开更多
目的探讨IFIT1(interferon-induced protein with tetratricopetide repeats 1)在放射损伤与感染应激中表达的特点和意义。方法采用5μg/mL的脂多糖(LPS)刺激Raw264.7、3T3和10T1/2细胞株5 h后,收集细胞,提取RNA,行RT-PCR观察IFIT1的表...目的探讨IFIT1(interferon-induced protein with tetratricopetide repeats 1)在放射损伤与感染应激中表达的特点和意义。方法采用5μg/mL的脂多糖(LPS)刺激Raw264.7、3T3和10T1/2细胞株5 h后,收集细胞,提取RNA,行RT-PCR观察IFIT1的表达情况,同时设定空白对照组。另外,将20只C57/BL6小鼠随机分为4组,采用12 Gray60Co一次性全身照射,分别于0 h、1 h、4 h和12 h脱颈处死取肝组织,用Western blot测定肝组织IFIT1表达变化。结果 Raw264.7、3T3和10T1/2细胞株经LPS刺激5 h后,均能诱导IFIT1表达,空白对照组表达为阴性。C57/BL6小鼠放射性损伤后,1 h IFIT1/Actin开始增高,12 h达最高,呈上升趋势,差异有极显著性统计学意义(P<0.01)。结论 LPS可刺激多种细胞株表达IFIT1,提示IFIT1可能参与创伤后毒血症的发生和发展;放射损伤早期能引起小鼠肝组织IFIT1的显著增高。展开更多
目的探讨干扰素诱导蛋白5(interferon-induced protein with tetratricopeptide repeats 5,IFIT5)在肝癌组织中的表达、临床意义及可能的分子机制。方法收集2013年在西安交通大学第一附属医院确诊的97例肝癌患者的肿瘤组织和癌旁组织。...目的探讨干扰素诱导蛋白5(interferon-induced protein with tetratricopeptide repeats 5,IFIT5)在肝癌组织中的表达、临床意义及可能的分子机制。方法收集2013年在西安交通大学第一附属医院确诊的97例肝癌患者的肿瘤组织和癌旁组织。实时定量PCR、Western blot及免疫组化检测IFIT5在肝癌及癌旁组织中的表达水平,并分析IFIT5表达水平与临床病理指标的关系及生存率。Transwell实验检测IFIT5对HCC细胞的侵袭及迁移作用,Western blot法检测IFIT5对HCC细胞EMT的影响。结果IFIT5在肝癌组织中表达水平显著高于相应癌旁组织(P<0.05);肝癌组织IFIT5高表达与TNM分期(P=0.018)及血管侵犯(P=0.009)显著相关;高表达IFIT5患者5年生存率明显高于低表达组(P<0.05);IFIT5可显著促进HCC细胞的侵袭及迁移(P<0.05);IFIT5促进HCC细胞EMT的发生(P<0.05)。结论IFIT5在肝癌组织中表达升高,其高表达与肝癌恶性临床病理特征有关,IFIT5能够通过调控HCC细胞EMT促进其侵袭和迁移。展开更多
为了筛选鸡IFIT5基因潜在的、具有生物学功能的nsSNPs(nonsense Single nucleotide polymorphisms,SNPs)位点和UTR-SNPs(untranslated regions-SNPs)位点。利用5种SNPs在线工具对nsSNPs进行分析预测有害位点;使用SWISS-MODEL,Pymol,Cons...为了筛选鸡IFIT5基因潜在的、具有生物学功能的nsSNPs(nonsense Single nucleotide polymorphisms,SNPs)位点和UTR-SNPs(untranslated regions-SNPs)位点。利用5种SNPs在线工具对nsSNPs进行分析预测有害位点;使用SWISS-MODEL,Pymol,Consurf server等软件对有害nsSNPs位点进行蛋白质的空间结构,氨基酸的氢键变化,及保守型等进行相关分析。此外,对UTR-SNPs位点使用UTRScan分析预测其结合模式元件是否改变。结果表明:从IFIT5基因的nsSNPs位点筛选出来的6个有害nsSNPs位点(L220I,L223M,L223V,Y375C,E391G和Y414D)都为潜在性功能位点,其中E391G位点最可能影响蛋白的结构和功能。从UTR-SNPs筛选出9个位于uORF的位点可能影响IFIT5基因的转录模式。展开更多
AIM To investigate the association between interferoninduced protein with tetratricopeptide repeats 1(IFIT1) polymorphisms and interferon-α(IFNα) treatment efficiency among Chinese hepatitis B virus(HBV) infection p...AIM To investigate the association between interferoninduced protein with tetratricopeptide repeats 1(IFIT1) polymorphisms and interferon-α(IFNα) treatment efficiency among Chinese hepatitis B virus(HBV) infection patients.METHODS Two hundred and twenty five newly diagnosed chronichepatitis B(CHB) patients were enrolled in the study. All of these patients received IFNα treatment for a course of 48 wk, and were followed up for 24 wk after the treatment was end. Clinical information about virological response, hepatitis B e antigen(HBe Ag) seroconversion rate and combined response at the end of the treatment, as well as the sustained response by the time of following up 24 wk after the treatment, was collected. Four tag-single nucleotide polymorphisms(SNPs) of IFIT1 were selected and assessed for their association with these clinical outcomes.RESULTS At the end of the treatment, HBe Ag seroconversion was observed in 27.1% patients. Thirty-six point nine percent patients achieved virological response, and 15.6% patients exhibited combined response. Sustained response was obtained in 26.2% patients. The main HBV genotype of the study was genotype B. Patients who infected with HBV genotype B or C showed better treatment efficiency, no matter which clinical outcome was considered. Among the four SNPs assessed, rs303218(A > G) was found to be significantly associated with the end point virological response when assuming additive model [OR = 0.64(95%CI: 0.42-0.96), P = 0.032]. Patients who carried rs303218 GG genotype had a rather higher rate of achieving virological response(response rate: 52%, OR = 0.40, 95%CI: 0.18-0.91; P = 0.028) when compared to those had AA genotype(response rate: 27%). The most significant interaction was observed in patients who had relative lower baseline aspartate transaminase. No association between SNPs and HBe Ag seroconversion, combined response or sustained response was observed.CONCLUSION IFIT1 involves in the regulation of IFNα treatment for CHB and its polymorphism rs303218 can predict the end point virological response. The finding requires further validation.展开更多
文摘目的获取小鼠IFIT1(Interferon-induced protein with tetratricopetide repeats 1)特异性多克隆抗体。方法扩增培养高效表达IFIT1的重组子pMAL-C2X-IFIT1,超声破碎后的细胞上清过Amylose Pre-packed column亲和层析柱,将所得纯化的融合蛋白MBP-IFIT1作为抗原,添加福氏佐机剂后免疫兔,收集抗血清,用Western blot和ELISA方法测定抗血清特异性反应和效价。结果纯化的MBP-IFIT1可诱导兔产生特异性免疫应答,所得抗体能够特异性识别融合蛋白中的IFIT1,ELISA结果显示其效价为1∶12800。结论MBP-IFIT1具有良好的抗原性,以该融合蛋白为抗原免疫兔成功制备出IFIT1特异性多克隆抗体。
文摘目的检测干扰素诱导蛋白IFIT1(interferon-induced protein with tetratricopetide repeats1)在小鼠各器官组织的表达状况,并在此基础上探讨颈交感神经阻滞(SB)对烧伤后早期小鼠肝脏IFIT1表达的影响。方法雄性C57/129小鼠50只,分为对照组和SB治疗组,TBSA15%～20%Ⅲ度小鼠烧伤模型,分别于烧伤前、烧伤后1、6、12h和24h取肝组织,采用免疫印迹法(Western blot)观察IFIT1的蛋白表达情况,同时提取正常小鼠心、肝、脾、肺、肾、小肠、淋巴细胞和骨骼肌组织RNA,以RT-PCR检测IFIT1表达情况。结果RT-PCR结果显示IFIT1在正常小鼠各组织均有表达,但组织间的表达量存在差异。Western blot结果表明对照组烧伤后肝组织IFIT1表达显著增高(P<0.01),烧伤后SB治疗组IFIT1表达量较对照组相应时相点降低非常显著(P<0.01)。结论IFIT1在小鼠体内广泛表达。SB对严重创伤的治疗作用可能是通过抑制肝内IFIT1表达来实现的。
文摘目的：观察小鼠心、肝、脾、肺、肾等器官干扰素诱导蛋白IFIT1（Interferon—induced protein with tetratfieopetide repeats1）的表达情况，探讨颈交感神经阻滞（cervical sympathetic block，SB）对烧伤后早期小鼠各器官IFIT1表达状况的影响．方法：将雄性昆明小鼠18只随机分为正常对照组、烧伤组和颈交感神经阻滞（SB）治疗组、烧伤组和SB治疗组采用体表总面积（Total body surface area，TBSA）15％-20％Ⅲ度小鼠烧伤模型，SB治疗组于烧伤后即刻进行双侧颈交感神经阻滞，各组分别于4h后取心、肝、脾、肺、肾组织，提取RNA，以RT—PCR检测IFIT1表达情况．结果：RT—PCR显示IFIT1在各组的各器官均有表达，并存在差异．烧伤组IFIT1在心、肝、脾表达低于正常组，同时在肺、肾表达高于正常组，差异显著（P〈0．01）．SB治疗组IFIT1的表达均低于正常组（P〈0．01）；同时，SB治疗组IFIT1在心、肝、肺、肾的表达也低于烧伤组（P〈0．01）．另外，在各组内的不同器官的IFIT1表达量存在差异（P〈0．05）．结论：烧伤后早期小鼠各器官IFIT1表达有不同的变化，可能与不同器官在烧伤应激反应过程中损伤程度不同有关．SB可显著降低各器官IFIT1的表达．
文摘目的：观察小鼠应激相关蛋白IFIT1（Interferon-induced protein with tetratricopetide repeats1）和JAB1（c-Jun平activation共处五项原则domain-binding protein 1）在肝脏的表达的细胞定位及氯胺酮对烧伤后早期肝脏IFIT1和JAB1表达的影响，初步探讨氯胺酮调节炎症反应的分子机制。方法：将15只健康C57／129小鼠随机分为3组（n=5），正常对照组、烧伤组、烧伤＋氯胺酮组。采用TBSA 15～20％Ⅲ度小鼠烧伤模型；氯胺酮10mg／kg，肌肉注射，烧伤15min给予。分别于烧伤后4h脱臼处死，取肝。采用免疫印迹法（Western blot）察各组肝IFIT1和JAB1的表达情况。取正常对照组肝组织作病理切片，行免疫组织化学方法对IFIT1和JAB1的表达做细胞定位。结果：烧伤组IFIT1表达较正常对照组显著增高（P〈0．01）．烧伤组JAB1表达较正常对照组显著降低（P〈0．01）：氯胺酮治疗组IFIT1表达较正常对照组、烧伤对照组显著降低（P〈0．01）．氯胺酮治疗组JAB1表达较正常对照组显著降低（P〈0．05）、较烧伤组显著升高（P〈0．05）。免疫组织化学方法显示．IFIT1在肝细胞胞浆表达为阳性．JAB1在肝细胞胞浆和胞核均表达为阳性。结论：烧伤早期小鼠肝组织IFIT1表达增高而JJAB1表达降低，氯胺酮可降低烧伤早期小鼠肝组织IFIT1的高表达，增高JAB1表达；氯胺酮可通过调控严重烧伤应激中IFIT1和JAB1的表达而在全身炎性反应综合症（Systemic inflammatory response syndrome SIRS）中发挥作用。
文摘目的探讨干扰素诱导蛋白5(interferon-induced protein with tetratricopeptide repeats 5,IFIT5)在肝癌组织中的表达、临床意义及可能的分子机制。方法收集2013年在西安交通大学第一附属医院确诊的97例肝癌患者的肿瘤组织和癌旁组织。实时定量PCR、Western blot及免疫组化检测IFIT5在肝癌及癌旁组织中的表达水平,并分析IFIT5表达水平与临床病理指标的关系及生存率。Transwell实验检测IFIT5对HCC细胞的侵袭及迁移作用,Western blot法检测IFIT5对HCC细胞EMT的影响。结果IFIT5在肝癌组织中表达水平显著高于相应癌旁组织(P<0.05);肝癌组织IFIT5高表达与TNM分期(P=0.018)及血管侵犯(P=0.009)显著相关;高表达IFIT5患者5年生存率明显高于低表达组(P<0.05);IFIT5可显著促进HCC细胞的侵袭及迁移(P<0.05);IFIT5促进HCC细胞EMT的发生(P<0.05)。结论IFIT5在肝癌组织中表达升高,其高表达与肝癌恶性临床病理特征有关,IFIT5能够通过调控HCC细胞EMT促进其侵袭和迁移。
文摘AIM To investigate the association between interferoninduced protein with tetratricopeptide repeats 1(IFIT1) polymorphisms and interferon-α(IFNα) treatment efficiency among Chinese hepatitis B virus(HBV) infection patients.METHODS Two hundred and twenty five newly diagnosed chronichepatitis B(CHB) patients were enrolled in the study. All of these patients received IFNα treatment for a course of 48 wk, and were followed up for 24 wk after the treatment was end. Clinical information about virological response, hepatitis B e antigen(HBe Ag) seroconversion rate and combined response at the end of the treatment, as well as the sustained response by the time of following up 24 wk after the treatment, was collected. Four tag-single nucleotide polymorphisms(SNPs) of IFIT1 were selected and assessed for their association with these clinical outcomes.RESULTS At the end of the treatment, HBe Ag seroconversion was observed in 27.1% patients. Thirty-six point nine percent patients achieved virological response, and 15.6% patients exhibited combined response. Sustained response was obtained in 26.2% patients. The main HBV genotype of the study was genotype B. Patients who infected with HBV genotype B or C showed better treatment efficiency, no matter which clinical outcome was considered. Among the four SNPs assessed, rs303218(A > G) was found to be significantly associated with the end point virological response when assuming additive model [OR = 0.64(95%CI: 0.42-0.96), P = 0.032]. Patients who carried rs303218 GG genotype had a rather higher rate of achieving virological response(response rate: 52%, OR = 0.40, 95%CI: 0.18-0.91; P = 0.028) when compared to those had AA genotype(response rate: 27%). The most significant interaction was observed in patients who had relative lower baseline aspartate transaminase. No association between SNPs and HBe Ag seroconversion, combined response or sustained response was observed.CONCLUSION IFIT1 involves in the regulation of IFNα treatment for CHB and its polymorphism rs303218 can predict the end point virological response. The finding requires further validation.
