目的研究麻杏解毒合剂对冠状病毒肺炎模型小鼠炎性因子表达的影响,基于p38MAPK/AP-1通路研究其疗效机制。方法60只昆明种(KM)小鼠随机分成空白对照组、模型组、p38MAPK抑制剂组及麻杏解毒合剂高、中、低剂量组,每组10只。采用模拟寒湿...目的研究麻杏解毒合剂对冠状病毒肺炎模型小鼠炎性因子表达的影响,基于p38MAPK/AP-1通路研究其疗效机制。方法60只昆明种(KM)小鼠随机分成空白对照组、模型组、p38MAPK抑制剂组及麻杏解毒合剂高、中、低剂量组,每组10只。采用模拟寒湿环境、表达h ACE2的重组腺相关病毒转导、SARS-CoV-2spike假病毒气管内给药建立小鼠冠状病毒肺炎模型。检测各组小鼠血清炎性因子、肺组织病理改变、肺组织p38MAPK、c-jun、c-fos的mRNA水平和蛋白表达情况。结果与空白对照组比较,模型组小鼠血清炎性因子白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、白细胞介素-8(IL-8)、肿瘤坏死因子-α(TNF-α)的表达均显著升高(P<0.01),肺组织炎性改变明显,c-fos m RNA水平和p-p38、c-fos、c-jun蛋白表达均显著增加(P<0.05)。与模型组比较,麻杏解毒合剂高、中剂量组小鼠血清炎性因子显著降低(P<0.01或P<0.05),小鼠肺组织炎性损伤明显减轻,同时肺组织中c-fos的mRNA水平和p-p38、c-fos的蛋白表达均显著下调(P<0.05)。结论麻杏解毒合剂能降低病毒性肺炎模型小鼠血清炎性因子水平,减轻肺组织炎性损伤,其疗效机制与抑制p38MAPK蛋白的磷酸化,下调AP-1通路mRNA及蛋白表达有关。展开更多
目的:构建Max二聚化蛋白1(Max dimerization protein 1,Mad1)的真核表达载体,研究Mad1对胃癌细胞增殖和迁移能力的影响。方法:利用DNA重组技术将Mad1基因克隆至pEGFP-N1载体,构建重组真核表达载体pEGFP-N1-Mad1。经酶切和测序鉴定后,采...目的:构建Max二聚化蛋白1(Max dimerization protein 1,Mad1)的真核表达载体,研究Mad1对胃癌细胞增殖和迁移能力的影响。方法:利用DNA重组技术将Mad1基因克隆至pEGFP-N1载体,构建重组真核表达载体pEGFP-N1-Mad1。经酶切和测序鉴定后,采用脂质体转染技术将重组质粒瞬时转染人胃癌AGS细胞,RT-PCR及Western blot检测Mad1基因和蛋白的表达。荧光显微镜观察Mad1在AGS细胞内的定位情况。CCK-8和Transwell实验研究Mad1对胃癌AGS细胞增殖和迁移能力的影响。结果:成功构建携带Mad1基因的真核表达载体pEGFP-N1-Mad1。将重组质粒瞬时转染AGS细胞后,RT-PCR和Western blot检测到Mad1基因和蛋白的表达。Mad1基因表达产物定位于AGS细胞核中。CCK-8和Transwell实验结果显示,转染Mad1的AGS细胞与转染空载体的AGS细胞及正常AGS细胞相比,细胞增殖活力和迁移能力明显降低。结论:成功构建了真核表达载体p EGFP-N1-Mad1,研究表明Mad1可以抑制胃癌AGS细胞的增殖和迁移。展开更多
Seed size is one of the vital traits determining seed appearance, quality, and yield. Untangling the genetic mechanisms regulating soybean 100-seed weight (100-SW), seed length and seed width across environments may p...Seed size is one of the vital traits determining seed appearance, quality, and yield. Untangling the genetic mechanisms regulating soybean 100-seed weight (100-SW), seed length and seed width across environments may provide a theoretical basis for improving seed yield. However, there are few reports related to QTL mapping of 100-SW across multiple ecological regions. In this study, 21 loci associated with seed size traits were identified using a genome-wide association of 5361 single nucleotide polymorphisms (SNPs) across three ecoregions in China, which could explain 8.12%–14.25% of the phenotypic variance respectively. A new locus, named as SW9-1 on chromosome 9 that explained 10.05%–10.93% of the seed weight variance was found significantly related to seed size traits, and was not previously reported. The selection effect analysis showed that SW9-1 locus has a relatively high phenotypic effect (13.67) on 100-SW, with a greater contribution by the accessions with bigger seeds (3.69) than the accessions with small seeds (1.66). Increases in seed weight were accompanied by increases in the frequency of SW9-1T allele, with >90% of the bred varieties with a 100-SW >30 g carrying SW9-1T. Analysis of SW9-1 allelic variation in additional soybean accessions showed that SW9-1T allele accounting for 13.83% of the wild accessions, while in 46.55% and 51.57% of the landraces and bred accessions, respectively, this results indicating that the SW9-1 locus has been subjected to artificial selection during the early stages of soybean breeding, especially the utilization of SW9-1T in edamame for big seed. These results suggest that SW9-1 is a novel and reliable locus associated with seed size traits, and might have an important implication for increasing soybean seed weight in molecular design breeding. Cloning this locus in future may provide new insights into the genetic mechanisms underlying soybean seed size traits.展开更多
文摘目的研究麻杏解毒合剂对冠状病毒肺炎模型小鼠炎性因子表达的影响,基于p38MAPK/AP-1通路研究其疗效机制。方法60只昆明种(KM)小鼠随机分成空白对照组、模型组、p38MAPK抑制剂组及麻杏解毒合剂高、中、低剂量组,每组10只。采用模拟寒湿环境、表达h ACE2的重组腺相关病毒转导、SARS-CoV-2spike假病毒气管内给药建立小鼠冠状病毒肺炎模型。检测各组小鼠血清炎性因子、肺组织病理改变、肺组织p38MAPK、c-jun、c-fos的mRNA水平和蛋白表达情况。结果与空白对照组比较,模型组小鼠血清炎性因子白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、白细胞介素-8(IL-8)、肿瘤坏死因子-α(TNF-α)的表达均显著升高(P<0.01),肺组织炎性改变明显,c-fos m RNA水平和p-p38、c-fos、c-jun蛋白表达均显著增加(P<0.05)。与模型组比较,麻杏解毒合剂高、中剂量组小鼠血清炎性因子显著降低(P<0.01或P<0.05),小鼠肺组织炎性损伤明显减轻,同时肺组织中c-fos的mRNA水平和p-p38、c-fos的蛋白表达均显著下调(P<0.05)。结论麻杏解毒合剂能降低病毒性肺炎模型小鼠血清炎性因子水平,减轻肺组织炎性损伤,其疗效机制与抑制p38MAPK蛋白的磷酸化,下调AP-1通路mRNA及蛋白表达有关。
基金supported by the National Key Research and Development Program of China (2016YFD0100201)the National Natural Science Foundation of China (31771819)+2 种基金the China Postdoctoral Science Foundation (2017M621990)the Introduced Leading Talent Research Team for Universities in Anhui Provincethe Natural Science Foundation of Anhui Province, China (1608085QC66)
文摘Seed size is one of the vital traits determining seed appearance, quality, and yield. Untangling the genetic mechanisms regulating soybean 100-seed weight (100-SW), seed length and seed width across environments may provide a theoretical basis for improving seed yield. However, there are few reports related to QTL mapping of 100-SW across multiple ecological regions. In this study, 21 loci associated with seed size traits were identified using a genome-wide association of 5361 single nucleotide polymorphisms (SNPs) across three ecoregions in China, which could explain 8.12%–14.25% of the phenotypic variance respectively. A new locus, named as SW9-1 on chromosome 9 that explained 10.05%–10.93% of the seed weight variance was found significantly related to seed size traits, and was not previously reported. The selection effect analysis showed that SW9-1 locus has a relatively high phenotypic effect (13.67) on 100-SW, with a greater contribution by the accessions with bigger seeds (3.69) than the accessions with small seeds (1.66). Increases in seed weight were accompanied by increases in the frequency of SW9-1T allele, with >90% of the bred varieties with a 100-SW >30 g carrying SW9-1T. Analysis of SW9-1 allelic variation in additional soybean accessions showed that SW9-1T allele accounting for 13.83% of the wild accessions, while in 46.55% and 51.57% of the landraces and bred accessions, respectively, this results indicating that the SW9-1 locus has been subjected to artificial selection during the early stages of soybean breeding, especially the utilization of SW9-1T in edamame for big seed. These results suggest that SW9-1 is a novel and reliable locus associated with seed size traits, and might have an important implication for increasing soybean seed weight in molecular design breeding. Cloning this locus in future may provide new insights into the genetic mechanisms underlying soybean seed size traits.
