取4月龄八眉猪和大白猪背部皮下脂肪组织,提取总RNA,根据大鼠细胞因子信号转导抑制因子-3基因(Suppressor Of Cytokine Signaling-3,SOCS-3)设计并合成引物,以猪-βactin基因作为内参,优化反应条件和体系,半定量(Semi Quantitative,SQ)R...取4月龄八眉猪和大白猪背部皮下脂肪组织,提取总RNA,根据大鼠细胞因子信号转导抑制因子-3基因(Suppressor Of Cytokine Signaling-3,SOCS-3)设计并合成引物,以猪-βactin基因作为内参,优化反应条件和体系,半定量(Semi Quantitative,SQ)RT-PCR单管扩增猪SOCS-3基因,经琼脂糖凝胶电泳分离后,Dolphin-DOC凝胶图像分析软件分析,测定各条带的光密度值,检测八眉猪和大白猪脂肪组织中SOCS-3基因mRNA的表达差异。结果表明:八眉猪脂肪组织SOCS-3 mRNA的表达丰度极显著高于大白猪(P<0.01)。这种差异可能是2种经济类型猪脂肪沉积能力不同的主要原因之一。展开更多
为了研究细胞因子信号转导分子3(suppressor of cytokine signals-3,SOCS-3)对造血发育的影响,构建了SOCS-3慢病毒siRNA干涉载体,并转染人红白血病细胞株K562.根据绿色荧光蛋白的表达进行流式分选后,获得了高表达慢病毒干涉载体的细胞....为了研究细胞因子信号转导分子3(suppressor of cytokine signals-3,SOCS-3)对造血发育的影响,构建了SOCS-3慢病毒siRNA干涉载体,并转染人红白血病细胞株K562.根据绿色荧光蛋白的表达进行流式分选后,获得了高表达慢病毒干涉载体的细胞.实时荧光定量PCR和Western-blot检测了转染细胞中SOCS-3基因的干涉效率,结果显示,与对照组相比,siRNA干涉后K562细胞SOCS-3基因的表达量仅为其相对表达量的22.1%,干涉效率77.9%;Western-blot结果显示,SOCS-3在蛋白质水平表达也明显受抑制.进一步对SOCS-3基因沉默后的K562细胞进行了诱导分化,并采用联苯胺染色法检测K562细胞向红系分化比例变化,免疫荧光染色检测细胞表面抗原的变化,RT-PCR检测造血相关基因的变化.结果发现,SOCS-3沉默后K562细胞向红系的发育能力显著提高.研究结果证明,SOCS-3在造血发育中有重要调控作用,而对其表达进行干涉或沉默将在规模化的红细胞诱导研究中发挥重要作用.展开更多
细胞因子是调节机体免疫和神经内分泌功能的生物活性物质，其信号的激发、放大和持续在时间和空间上都受到严格调控 。细胞因子信号传导抑制因子（Suppressor of cytokine signaling．socs）是细胞因子信号通路的负调节因子，通过负反...细胞因子是调节机体免疫和神经内分泌功能的生物活性物质，其信号的激发、放大和持续在时间和空间上都受到严格调控 。细胞因子信号传导抑制因子（Suppressor of cytokine signaling．socs）是细胞因子信号通路的负调节因子，通过负反馈抑制细胞因子的信号传递，防止过度的信号反应干扰机体代谢平衡和细胞功能 .展开更多
从中国地方猪品种八眉猪(BaMei)肾脏组织中提取总RNA,采用RT-PCR方法克隆了猪SOCS-2(suppressor of cytokinesignaling-2,细胞因子信号转导抑制因子-2)基因的cDNA序列,经T/A克隆,插入到pMD19-T载体上,导入大肠杆菌DH-5α,阳性克隆经PCR...从中国地方猪品种八眉猪(BaMei)肾脏组织中提取总RNA,采用RT-PCR方法克隆了猪SOCS-2(suppressor of cytokinesignaling-2,细胞因子信号转导抑制因子-2)基因的cDNA序列,经T/A克隆,插入到pMD19-T载体上,导入大肠杆菌DH-5α,阳性克隆经PCR鉴定后进行测序,将测序结果与GenBank中已登录的人、大鼠和小鼠SOCS-2基因的序列进行同源性比较,利用生物信息学和分子生物学软件对猪SOCS-2基因编码的蛋白进行结构预测。结果表明:首次成功克隆了猪SOCS-2基因的cDNA序列(GenBank登录号为EF121242),其长度为822bp,该基因ORF区核苷酸序列与其他物种相比同源性达到93%以上,氨基酸同源性则达到89%以上,生物信息学分析表明该蛋白分子量为22.25kD,等电点pI=8.30,包含199个氨基酸残基。该基因cDNA序列的克隆,有利于进一步研究SOCS-2调节机体发育的分子机理。展开更多
为探究团头鲂细胞因子信号转导抑制因子(suppressor of cytokine signaling,SOCS)家族基因的序列特征和功能,从NCBI数据库获取socs1、socs2、socs3a、socs3b、socs4、socs5a、socs5b、socs6、socs7、socs9共10个socs基因的cDNA序列,利用...为探究团头鲂细胞因子信号转导抑制因子(suppressor of cytokine signaling,SOCS)家族基因的序列特征和功能,从NCBI数据库获取socs1、socs2、socs3a、socs3b、socs4、socs5a、socs5b、socs6、socs7、socs9共10个socs基因的cDNA序列,利用ExPASy、SMART等在线网站对其进行生物信息学分析,预测团头鲂10个SOCS的理论分子质量、理论等电点与结构域等分子特性。系统进化(MEGA 6软件)分析结果显示,10个SOCS可分为2个亚族:Ⅰ型亚族包括SOCS4、SOCS5a、SOCS5b、SOCS6、SOCS7与SOCS9,Ⅱ型亚族包括SOCS1、SOCS2、SOCS3a与SOCS3b。半定量PCR结果显示,健康团头鲂10个socs基因在被检测组织中具有明显的组织特异性,socs1、socs2、socs3在多个组织中表达量较高。在嗜水气单胞菌感染后,采用实时定量PCR检测团头鲂socs基因在脾脏、体肾、头肾中的表达量,其中socs1、socs2、socs3a、socs3b表达量均显著上调。结果表明,团头鲂SOCS家族基因的表达模式各不相同,预示其功能的多样性,其中socs1、socs2、socs3在免疫应答中发挥重要作用。展开更多
BACKGROUND Chest wall tuberculosis(TB)and triple-negative essential thrombocythemia(TNET)are rare medical conditions,and their combination is extremely rare globally.Only one case of TB peritonitis with thrombocytosis...BACKGROUND Chest wall tuberculosis(TB)and triple-negative essential thrombocythemia(TNET)are rare medical conditions,and their combination is extremely rare globally.Only one case of TB peritonitis with thrombocytosis has been reported,which was identified in 1974.CASE SUMMARY Herein,we report the case of a 23-year-old man with concurrent chest wall mass and TN-ET.The patient presented to a local hospital due to having a headache and low-grade fever for 2 d,with their bodily temperature fluctuating at around 36.8°C.Hematological analysis showed a high platelet count of 1503×109/L.Subsequently,the patient visited our hospital for further investigation.Computed tomography of the chest suggested a submural soft tissue density shadow in the left lower chest wall.After surgical resection,the pathological findings of the swelling were reported as TB with massive caseous necrosis.According to the World Health Organization diagnostic criteria,the patient was diagnosed with TN-ET,as they met the requirement of four main criteria or the first three main criteria and one secondary criterion.The patient was eventually diagnosed with chest wall TB with TN-ET,which is extremely rare.CONCLUSION Chest wall TB is rare.TN-ET diagnosis requires secondary factor exclusion and satisfaction of primary diagnostic criteria.miRNA,combined with the methylation process,could explain suppressor of cytokine signaling(SOCS)1 and SOCS3 downregulation in ET-JAK2V617F-negative patients.The miRNA could participate in JAK2 pathway activation.SOCS3 may be a novel MPN biomarker.展开更多
文摘为了研究细胞因子信号转导分子3(suppressor of cytokine signals-3,SOCS-3)对造血发育的影响,构建了SOCS-3慢病毒siRNA干涉载体,并转染人红白血病细胞株K562.根据绿色荧光蛋白的表达进行流式分选后,获得了高表达慢病毒干涉载体的细胞.实时荧光定量PCR和Western-blot检测了转染细胞中SOCS-3基因的干涉效率,结果显示,与对照组相比,siRNA干涉后K562细胞SOCS-3基因的表达量仅为其相对表达量的22.1%,干涉效率77.9%;Western-blot结果显示,SOCS-3在蛋白质水平表达也明显受抑制.进一步对SOCS-3基因沉默后的K562细胞进行了诱导分化,并采用联苯胺染色法检测K562细胞向红系分化比例变化,免疫荧光染色检测细胞表面抗原的变化,RT-PCR检测造血相关基因的变化.结果发现,SOCS-3沉默后K562细胞向红系的发育能力显著提高.研究结果证明,SOCS-3在造血发育中有重要调控作用,而对其表达进行干涉或沉默将在规模化的红细胞诱导研究中发挥重要作用.
文摘从中国地方猪品种八眉猪(BaMei)肾脏组织中提取总RNA,采用RT-PCR方法克隆了猪SOCS-2(suppressor of cytokinesignaling-2,细胞因子信号转导抑制因子-2)基因的cDNA序列,经T/A克隆,插入到pMD19-T载体上,导入大肠杆菌DH-5α,阳性克隆经PCR鉴定后进行测序,将测序结果与GenBank中已登录的人、大鼠和小鼠SOCS-2基因的序列进行同源性比较,利用生物信息学和分子生物学软件对猪SOCS-2基因编码的蛋白进行结构预测。结果表明:首次成功克隆了猪SOCS-2基因的cDNA序列(GenBank登录号为EF121242),其长度为822bp,该基因ORF区核苷酸序列与其他物种相比同源性达到93%以上,氨基酸同源性则达到89%以上,生物信息学分析表明该蛋白分子量为22.25kD,等电点pI=8.30,包含199个氨基酸残基。该基因cDNA序列的克隆,有利于进一步研究SOCS-2调节机体发育的分子机理。
文摘为探究团头鲂细胞因子信号转导抑制因子(suppressor of cytokine signaling,SOCS)家族基因的序列特征和功能,从NCBI数据库获取socs1、socs2、socs3a、socs3b、socs4、socs5a、socs5b、socs6、socs7、socs9共10个socs基因的cDNA序列,利用ExPASy、SMART等在线网站对其进行生物信息学分析,预测团头鲂10个SOCS的理论分子质量、理论等电点与结构域等分子特性。系统进化(MEGA 6软件)分析结果显示,10个SOCS可分为2个亚族:Ⅰ型亚族包括SOCS4、SOCS5a、SOCS5b、SOCS6、SOCS7与SOCS9,Ⅱ型亚族包括SOCS1、SOCS2、SOCS3a与SOCS3b。半定量PCR结果显示,健康团头鲂10个socs基因在被检测组织中具有明显的组织特异性,socs1、socs2、socs3在多个组织中表达量较高。在嗜水气单胞菌感染后,采用实时定量PCR检测团头鲂socs基因在脾脏、体肾、头肾中的表达量,其中socs1、socs2、socs3a、socs3b表达量均显著上调。结果表明,团头鲂SOCS家族基因的表达模式各不相同,预示其功能的多样性,其中socs1、socs2、socs3在免疫应答中发挥重要作用。
文摘BACKGROUND Chest wall tuberculosis(TB)and triple-negative essential thrombocythemia(TNET)are rare medical conditions,and their combination is extremely rare globally.Only one case of TB peritonitis with thrombocytosis has been reported,which was identified in 1974.CASE SUMMARY Herein,we report the case of a 23-year-old man with concurrent chest wall mass and TN-ET.The patient presented to a local hospital due to having a headache and low-grade fever for 2 d,with their bodily temperature fluctuating at around 36.8°C.Hematological analysis showed a high platelet count of 1503×109/L.Subsequently,the patient visited our hospital for further investigation.Computed tomography of the chest suggested a submural soft tissue density shadow in the left lower chest wall.After surgical resection,the pathological findings of the swelling were reported as TB with massive caseous necrosis.According to the World Health Organization diagnostic criteria,the patient was diagnosed with TN-ET,as they met the requirement of four main criteria or the first three main criteria and one secondary criterion.The patient was eventually diagnosed with chest wall TB with TN-ET,which is extremely rare.CONCLUSION Chest wall TB is rare.TN-ET diagnosis requires secondary factor exclusion and satisfaction of primary diagnostic criteria.miRNA,combined with the methylation process,could explain suppressor of cytokine signaling(SOCS)1 and SOCS3 downregulation in ET-JAK2V617F-negative patients.The miRNA could participate in JAK2 pathway activation.SOCS3 may be a novel MPN biomarker.
