泛素受体蛋白DSK2(dominant suppressor of KAR2)在植物生长发育和逆境胁迫中发挥重要作用,但其在水稻抗病和渗透胁迫中的作用尚未见报道。本研究发现OsDSK2b受多种逆境的调控,该基因的表达水平在稻瘟病菌侵染和20%PEG-6000处理后显著...泛素受体蛋白DSK2(dominant suppressor of KAR2)在植物生长发育和逆境胁迫中发挥重要作用,但其在水稻抗病和渗透胁迫中的作用尚未见报道。本研究发现OsDSK2b受多种逆境的调控,该基因的表达水平在稻瘟病菌侵染和20%PEG-6000处理后显著降低。时空表达分析表明OsDSK2b基因在三叶期幼苗中的表达水平最高,亚细胞定位结果显示该蛋白定位于细胞质。接种稻瘟病菌(GD08-T13和Guy11)后,敲除植株的病斑面积约为0.05 cm^(2)和0.10~0.13 cm^(2),远小于野生型植株的病斑面积(0.24 cm^(2)和0.31 cm^(2))。敲除OsDSK2b显著增强水稻的叶瘟抗性。而且在病原菌侵染后,敲除植株中病程相关蛋白(PR)基因的表达受到明显诱导。敲除OsDSK2b也显著增强水稻的渗透胁迫抗性,20%PEG-6000模拟渗透胁迫处理后,OsDSK2b敲除植株的存活率为58.3%~74.0%,显著高于野生型植株的存活率(17.0%)。OsDSK2b敲除植株的离子渗透率和植株失水率则显著低于野生型植株。扫描显微镜的结果表明,无论在20%PEG-6000处理前后,敲除OsDSK2b能够促进气孔的闭合,且在20%PEG-6000处理后,这种促进作用会更强。此外,qRT-PCR结果表明,在20%PEG-6000处理后,OsDSK2b敲除植株中DREB基因以及脱落酸(ABA)合成或通路相关基因的表达水平显著高于野生型植株,且敲除植株ko6和ko14的内源ABA含量分别为314.2 ng g^(-1)和344.4 ng g^(-1),显著高于野生型植株的内源ABA含量(206.8 ng g^(-1)),揭示OsDSK2b参与调控渗透胁迫的过程同时涉及ABA依赖和ABA非依赖途径。本研究通过解析OsDSK2b基因在水稻应对生物与非生物胁迫时所发挥的调控作用,为水稻抗性品种的选育提供了新的候选基因。展开更多
目的:评价日常活动和手术应激评估(Estimation of physiologic ability and surgical stress,E-PASS)系统用于评估老龄患者消化道手术后并发症和转归的临床价值。方法:回顾性分析2011年7月至2013年7月西京医院消化外科所有65岁以上的患...目的:评价日常活动和手术应激评估(Estimation of physiologic ability and surgical stress,E-PASS)系统用于评估老龄患者消化道手术后并发症和转归的临床价值。方法:回顾性分析2011年7月至2013年7月西京医院消化外科所有65岁以上的患者的临床资料,计算其中行消化道手术者的E-PASS评分,并记录这些患者术后并发症的发生情况和患者术后的住院时间。分析E-PASS评分和几项该评分未涉及的因素与老龄患者消化道手术后并发症的发病率、死亡率、住院时间的相关性。结果:研究共纳入1236例老龄行消化道手术的患者,其中521例发生术后并发症(42.15%),8例死亡(0.65%)。患者术前E-PASS评分系统中,三项评分均与术后住院时间相关,术前风险评分(Preoperative risk score,PRS)和综合风险评分(Comprehensive risk score,CRS)与术后并发症的发病率和死亡率显著相关(P均<0.05)。E-PASS评分系统未包含的指标中,麻醉方法与术后并发症发生和住院时间无关,术后入ICU、术中使用血管活性药物和急诊手术与术后发病率、死亡率和住院时间相关(P均<0.05)。结论:E-PASS评分系统可用于预测老龄患者行消化道手术后并发症的发生情况和转归,纳入术后入ICU、术中使用血管活性药物和急诊手术三项指标可能进一步提高E-PASS评分系统的预测准确性。展开更多
GGGGCC repeat expansions in the C9 ORF72 gene are the most common cause of amyotrophic lateral sclerosis and frontotemporal dementia(c9 ALS/FTD). It has been reported that hexanucleotide repeat expansions in C9 ORF72 ...GGGGCC repeat expansions in the C9 ORF72 gene are the most common cause of amyotrophic lateral sclerosis and frontotemporal dementia(c9 ALS/FTD). It has been reported that hexanucleotide repeat expansions in C9 ORF72 produce five dipeptide repeat(DPR) proteins by an unconventional repeat-associated non-ATG(RAN)translation. Within the five DPR proteins, poly-PR and poly-GR that contain arginine are more toxic than the other DPRs(poly-GA, poly-GP, and poly-PA). Here, we demonstrated that poly-PR peptides transferred into cells by endocytosis in a clathrin-dependent manner, leading to endoplasmic reticulum stress and cell death. In SH-SY5 Y cells and primary cortical neurons, poly-PR activated JUN amino-terminal kinase(JNK) and increased the levels of p53 and Bax. The uptake of poly-PR peptides by cells was significantly inhibited by knockdown of clathrin or by chlorpromazine, an inhibitor that blocks clathrin-mediated endocytosis. Inhibition of clathrin-dependent endocytosis by chlorpromazine significantly blocked the transfer of poly-PR peptides into cells, and attenuated poly-PRinduced JNK activation and cell death. Our data revealed that the uptake of poly-PR undergoes clathrin-dependentendocytosis and blockade of this process prevents the toxic effects of synthetic poly-PR peptides.展开更多
文摘泛素受体蛋白DSK2(dominant suppressor of KAR2)在植物生长发育和逆境胁迫中发挥重要作用,但其在水稻抗病和渗透胁迫中的作用尚未见报道。本研究发现OsDSK2b受多种逆境的调控,该基因的表达水平在稻瘟病菌侵染和20%PEG-6000处理后显著降低。时空表达分析表明OsDSK2b基因在三叶期幼苗中的表达水平最高,亚细胞定位结果显示该蛋白定位于细胞质。接种稻瘟病菌(GD08-T13和Guy11)后,敲除植株的病斑面积约为0.05 cm^(2)和0.10~0.13 cm^(2),远小于野生型植株的病斑面积(0.24 cm^(2)和0.31 cm^(2))。敲除OsDSK2b显著增强水稻的叶瘟抗性。而且在病原菌侵染后,敲除植株中病程相关蛋白(PR)基因的表达受到明显诱导。敲除OsDSK2b也显著增强水稻的渗透胁迫抗性,20%PEG-6000模拟渗透胁迫处理后,OsDSK2b敲除植株的存活率为58.3%~74.0%,显著高于野生型植株的存活率(17.0%)。OsDSK2b敲除植株的离子渗透率和植株失水率则显著低于野生型植株。扫描显微镜的结果表明,无论在20%PEG-6000处理前后,敲除OsDSK2b能够促进气孔的闭合,且在20%PEG-6000处理后,这种促进作用会更强。此外,qRT-PCR结果表明,在20%PEG-6000处理后,OsDSK2b敲除植株中DREB基因以及脱落酸(ABA)合成或通路相关基因的表达水平显著高于野生型植株,且敲除植株ko6和ko14的内源ABA含量分别为314.2 ng g^(-1)和344.4 ng g^(-1),显著高于野生型植株的内源ABA含量(206.8 ng g^(-1)),揭示OsDSK2b参与调控渗透胁迫的过程同时涉及ABA依赖和ABA非依赖途径。本研究通过解析OsDSK2b基因在水稻应对生物与非生物胁迫时所发挥的调控作用,为水稻抗性品种的选育提供了新的候选基因。
基金supported by the National Natural Science Foundation of China (81761148024 and 31871023)the National Key Scientific R&D Program of China (2016YFC1306000)+1 种基金Suzhou Clinical Research Center of Neurological Disease (Szzx201503)a Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions, China
文摘GGGGCC repeat expansions in the C9 ORF72 gene are the most common cause of amyotrophic lateral sclerosis and frontotemporal dementia(c9 ALS/FTD). It has been reported that hexanucleotide repeat expansions in C9 ORF72 produce five dipeptide repeat(DPR) proteins by an unconventional repeat-associated non-ATG(RAN)translation. Within the five DPR proteins, poly-PR and poly-GR that contain arginine are more toxic than the other DPRs(poly-GA, poly-GP, and poly-PA). Here, we demonstrated that poly-PR peptides transferred into cells by endocytosis in a clathrin-dependent manner, leading to endoplasmic reticulum stress and cell death. In SH-SY5 Y cells and primary cortical neurons, poly-PR activated JUN amino-terminal kinase(JNK) and increased the levels of p53 and Bax. The uptake of poly-PR peptides by cells was significantly inhibited by knockdown of clathrin or by chlorpromazine, an inhibitor that blocks clathrin-mediated endocytosis. Inhibition of clathrin-dependent endocytosis by chlorpromazine significantly blocked the transfer of poly-PR peptides into cells, and attenuated poly-PRinduced JNK activation and cell death. Our data revealed that the uptake of poly-PR undergoes clathrin-dependentendocytosis and blockade of this process prevents the toxic effects of synthetic poly-PR peptides.
