目的探讨益气活血化浊解毒方调控KAT3B/STING轴介导的小胶质细胞极化对缺血性脑卒中大鼠神经功能的影响。方法复制缺血性脑卒中大鼠模型。将复制成功的60只大鼠分为缺血性脑卒中组、益气活血化浊解毒方低剂量组(低剂量组)、益气活血化...目的探讨益气活血化浊解毒方调控KAT3B/STING轴介导的小胶质细胞极化对缺血性脑卒中大鼠神经功能的影响。方法复制缺血性脑卒中大鼠模型。将复制成功的60只大鼠分为缺血性脑卒中组、益气活血化浊解毒方低剂量组(低剂量组)、益气活血化浊解毒方高剂量组(高剂量组)、益气活血化浊解毒方高剂量+2,5-己酮可可碱(DMXAA)组(高剂量+DMXAA组),每组15只。另取15只正常大鼠为Sham组。低剂量组大鼠灌胃0.2 mL 27.5 g/mL的益气活血化浊解毒方药液,高剂量组大鼠灌胃0.2 mL 55.0 g/mL的益气活血化浊解毒方药液,高剂量+DMXAA组大鼠灌胃0.2 mL 55.0 g/mL的益气活血化浊解毒方药液和25 mg/kg的DMXAA,Sham组和缺血性脑卒中组给予等体积生理盐水代替药物;除高剂量+DMXAA组外,其余组大鼠再给予等体积二甲基亚砜溶液灌胃,1 d/次,连续4周。Zea-Longa评分和网屏试验评分评估大鼠神经功能和神经行为学;检测大鼠脑组织含水量;TTC染色法检测大鼠脑梗死面积;酶联免疫吸附试验检测大鼠血清LDH和NGF水平和缺血侧皮层组织TNF-α、IL-6、IL-10水平;HE染色观察大鼠缺血侧皮层组织病理变化;Western blotting检测大鼠缺血侧皮层组织iNOS、Iba1、CD68、CD40、CD206、Arg-1、Cleaved caspase-3、KAT3B、STING蛋白表达。结果缺血性脑卒中组大鼠Zea-Longa评分和网屏试验评分、脑组织含水量、脑梗死面积百分比均高于Sham组(P<0.05),低剂量组和高剂量组大鼠Zea-Longa评分和网屏试验评分、脑组织含水量、脑梗死面积百分比均低于缺血性脑卒中组(P<0.05),高剂量+DMXAA组大鼠Zea-Longa评分和网屏试验评分、脑组织含水量、脑梗死面积百分比均高于高剂量组(P<0.05)。缺血性脑卒中组大鼠NGF和IL-10水平低于Sham组,LDH、TNF-α和IL-6水平高于Sham组(P<0.05);低剂量组和高剂量组大鼠NGF和IL-10水平高于缺血性脑卒中组,LDH、TNF-α和IL-6水平低于缺血性脑卒中组(P<0.05);高剂量+DMXAA组大鼠NGF和IL-10水平低于高剂量组,LDH、TNF-α和IL-6高于高剂量组(P<0.05)。与缺血性脑卒中组比较,低剂量组和高剂量组缺血侧皮层神经元形态有明显改善。缺血性脑卒中组大鼠缺血侧皮层组织中iNOS、Iba1、CD68和CD40蛋白相对表达量比Sham组高,CD206和Arg-1蛋白相对表达量比Sham组低(P<0.05);低剂量组和高剂量组大鼠缺血侧皮层组织中iNOS、Iba1、CD68和CD40蛋白相对表达量比缺血性脑卒中组低,CD206和Arg-1蛋白相对表达量比缺血性脑卒中组高(P<0.05);高剂量+DMXAA组大鼠缺血侧皮层组织中iNOS、Iba1、CD68和CD40蛋白相对表达量比高剂量组高,CD206和Arg-1蛋白相对表达量比高剂量组低(P<0.05)。结论益气活血化浊解毒方可调节缺血性脑卒中大鼠小胶质细胞极化,减轻神经炎症和神经损伤,改善神经功能,可能与抑制KAT3B/STING轴有关。展开更多
BACKGROUND Gastric cancer ranks among the leading malignancies worldwide,noted for its high morbidity and mortality,and remains a significant challenge to global public health.AIM To investigate the association betwee...BACKGROUND Gastric cancer ranks among the leading malignancies worldwide,noted for its high morbidity and mortality,and remains a significant challenge to global public health.AIM To investigate the association between the expression of splicing factor 3b subunit 4(SF3B4)and high mobility group box 1(HMGB1)with the clinical characteristics and prognostic outcomes of gastric cancer patients.METHODS A retrospective cohort study was conducted involving 114 individuals diagnosed with gastric cancer and admitted to our institution from January 2020 to December 2021.A comparison group of 90 patients diagnosed with benign gastric disorders during the same period was also included.Expression levels of SF3B4 and HMGB1 were assessed using real-time quantitative polymerase chain reaction.Expression patterns were analyzed in relation to various clinicopathological features.Receiver operating characteristic curves were constructed to evaluate the ability of SF3B4 and HMGB1,alone and in combination,to predict unfavorable one-year outcomes.Multivariate logistic regression was utilized to identify independent predictors of mortality.Kaplan-Meier survival curves were generated to examine survival differences based on SF3B4 and HMGB1 expression levels.RESULTS Both SF3B4 and HMGB1 were markedly upregulated in tumor tissues of gastric cancer patients compared to adjacent normal tissues and to tissues from nonmalignant gastric disease patients(^(a)P<0.05).Higher expression levels of these two genes were significantly associated with aggressive pathological features,including poor differentiation,tumor size>5 cm,deep infiltration(T3-T4),lymph node involvement,and advanced clinical stage(III–IV)(^(a)P<0.05).Receiver operating characteristic analysis revealed that the combined use of SF3B4 and HMGB1 yielded an area under the curve of 0.914,surpassing the predictive performance of either marker alone(SF3B4:0.776;HMGB1:0.757).Multivariate analysis identified SF3B4≥1.45,HMGB1≥0.93,poor differentiation,larger tumor size,deeper invasion,lymph node metastasis,and advanced clinical tumor-node-metastasis staging as independent factors contributing to one-year mortality(^(a)P<0.05).Survival analysis indicated that patients with elevated SF3B4 and HMGB1 levels had a shorter median survival(25.74±5.46 months)compared to those with lower expression levels(33.29±6.71 months,log-rank=10.534,^(a)P<0.05).CONCLUSION Elevated SF3B4 and HMGB1 expression in gastric cancer tissue is significantly associated with tumor aggressiveness,worse prognosis,and reduced survival.These biomarkers may offer clinical value in stratifying patients by risk and in forecasting outcomes.Their combined assessment improves predictive accuracy for poor prognosis and may serve as a more effective tool than individual evaluation.展开更多
目的:探讨敲低小鼠胚胎腭突间充质细胞(mEPMCs)初级纤毛(PC)重要组件驱动蛋白家族成员3B(KIF3B)基因对细胞自噬水平的作用,并阐明其作用机制。方法:收集体外培养孕龄14.5 d C57BL/6J小鼠的mEPMCs,根据是否敲低KIF3B基因和使用平滑因子...目的:探讨敲低小鼠胚胎腭突间充质细胞(mEPMCs)初级纤毛(PC)重要组件驱动蛋白家族成员3B(KIF3B)基因对细胞自噬水平的作用,并阐明其作用机制。方法:收集体外培养孕龄14.5 d C57BL/6J小鼠的mEPMCs,根据是否敲低KIF3B基因和使用平滑因子受体激动剂(SAG)激活音猬因子(Shh)信号通路及其下游共受体Smo,分为对照组(给予生理盐水)、空载病毒转染细胞组(sh-NC组)(给予慢病毒转染)、敲低KIF3B组(sh-KIF3B组)(给予KIF3B基因敲低)、敲低KIF3B加入SAG组(sh-KIF3B+SAG组)(给予KIF3B基因敲低后加入SAG),每组5只大鼠。透射电镜观察各组mEPMCs中自噬小体/自噬溶酶体形态表现及数量,Western blotting法检测各组mEPMCs中自噬相关蛋白苄氯素1(Beclin-1)和p62及Shh信号通路中Shh和Smo蛋白表达水平。结果:透射电镜观察,与对照组比较,sh-KIF3B组mEPMCs中自噬小体/自噬溶酶体数量明显增加(P<0.05);与sh-KIF3B组比较,sh-KIF3B+SAG组mEPMCs中自噬小体/自噬溶酶体数量明显减少(P<0.05)。Western blotting法检测,与对照组比较,sh-KIF3B组mEPMCs中Beclin-1蛋白表达水平明显升高(P<0.05),KIF3B、p62、Shh和Smo蛋白表达水平均明显降低(P<0.01);与sh-KIF3B组比较,sh-KIF3B+SAG组mEPMCs中Shh、Smo和p62蛋白表达水平明显升高(P<0.01),Beclin-1蛋白表达水平明显降低(P<0.01)。结论:敲低KIF3B基因可促进mEPMCs自噬,其机制可能与其抑制Shh信号通路有关。展开更多
文摘目的探讨益气活血化浊解毒方调控KAT3B/STING轴介导的小胶质细胞极化对缺血性脑卒中大鼠神经功能的影响。方法复制缺血性脑卒中大鼠模型。将复制成功的60只大鼠分为缺血性脑卒中组、益气活血化浊解毒方低剂量组(低剂量组)、益气活血化浊解毒方高剂量组(高剂量组)、益气活血化浊解毒方高剂量+2,5-己酮可可碱(DMXAA)组(高剂量+DMXAA组),每组15只。另取15只正常大鼠为Sham组。低剂量组大鼠灌胃0.2 mL 27.5 g/mL的益气活血化浊解毒方药液,高剂量组大鼠灌胃0.2 mL 55.0 g/mL的益气活血化浊解毒方药液,高剂量+DMXAA组大鼠灌胃0.2 mL 55.0 g/mL的益气活血化浊解毒方药液和25 mg/kg的DMXAA,Sham组和缺血性脑卒中组给予等体积生理盐水代替药物;除高剂量+DMXAA组外,其余组大鼠再给予等体积二甲基亚砜溶液灌胃,1 d/次,连续4周。Zea-Longa评分和网屏试验评分评估大鼠神经功能和神经行为学;检测大鼠脑组织含水量;TTC染色法检测大鼠脑梗死面积;酶联免疫吸附试验检测大鼠血清LDH和NGF水平和缺血侧皮层组织TNF-α、IL-6、IL-10水平;HE染色观察大鼠缺血侧皮层组织病理变化;Western blotting检测大鼠缺血侧皮层组织iNOS、Iba1、CD68、CD40、CD206、Arg-1、Cleaved caspase-3、KAT3B、STING蛋白表达。结果缺血性脑卒中组大鼠Zea-Longa评分和网屏试验评分、脑组织含水量、脑梗死面积百分比均高于Sham组(P<0.05),低剂量组和高剂量组大鼠Zea-Longa评分和网屏试验评分、脑组织含水量、脑梗死面积百分比均低于缺血性脑卒中组(P<0.05),高剂量+DMXAA组大鼠Zea-Longa评分和网屏试验评分、脑组织含水量、脑梗死面积百分比均高于高剂量组(P<0.05)。缺血性脑卒中组大鼠NGF和IL-10水平低于Sham组,LDH、TNF-α和IL-6水平高于Sham组(P<0.05);低剂量组和高剂量组大鼠NGF和IL-10水平高于缺血性脑卒中组,LDH、TNF-α和IL-6水平低于缺血性脑卒中组(P<0.05);高剂量+DMXAA组大鼠NGF和IL-10水平低于高剂量组,LDH、TNF-α和IL-6高于高剂量组(P<0.05)。与缺血性脑卒中组比较,低剂量组和高剂量组缺血侧皮层神经元形态有明显改善。缺血性脑卒中组大鼠缺血侧皮层组织中iNOS、Iba1、CD68和CD40蛋白相对表达量比Sham组高,CD206和Arg-1蛋白相对表达量比Sham组低(P<0.05);低剂量组和高剂量组大鼠缺血侧皮层组织中iNOS、Iba1、CD68和CD40蛋白相对表达量比缺血性脑卒中组低,CD206和Arg-1蛋白相对表达量比缺血性脑卒中组高(P<0.05);高剂量+DMXAA组大鼠缺血侧皮层组织中iNOS、Iba1、CD68和CD40蛋白相对表达量比高剂量组高,CD206和Arg-1蛋白相对表达量比高剂量组低(P<0.05)。结论益气活血化浊解毒方可调节缺血性脑卒中大鼠小胶质细胞极化,减轻神经炎症和神经损伤,改善神经功能,可能与抑制KAT3B/STING轴有关。
文摘BACKGROUND Gastric cancer ranks among the leading malignancies worldwide,noted for its high morbidity and mortality,and remains a significant challenge to global public health.AIM To investigate the association between the expression of splicing factor 3b subunit 4(SF3B4)and high mobility group box 1(HMGB1)with the clinical characteristics and prognostic outcomes of gastric cancer patients.METHODS A retrospective cohort study was conducted involving 114 individuals diagnosed with gastric cancer and admitted to our institution from January 2020 to December 2021.A comparison group of 90 patients diagnosed with benign gastric disorders during the same period was also included.Expression levels of SF3B4 and HMGB1 were assessed using real-time quantitative polymerase chain reaction.Expression patterns were analyzed in relation to various clinicopathological features.Receiver operating characteristic curves were constructed to evaluate the ability of SF3B4 and HMGB1,alone and in combination,to predict unfavorable one-year outcomes.Multivariate logistic regression was utilized to identify independent predictors of mortality.Kaplan-Meier survival curves were generated to examine survival differences based on SF3B4 and HMGB1 expression levels.RESULTS Both SF3B4 and HMGB1 were markedly upregulated in tumor tissues of gastric cancer patients compared to adjacent normal tissues and to tissues from nonmalignant gastric disease patients(^(a)P<0.05).Higher expression levels of these two genes were significantly associated with aggressive pathological features,including poor differentiation,tumor size>5 cm,deep infiltration(T3-T4),lymph node involvement,and advanced clinical stage(III–IV)(^(a)P<0.05).Receiver operating characteristic analysis revealed that the combined use of SF3B4 and HMGB1 yielded an area under the curve of 0.914,surpassing the predictive performance of either marker alone(SF3B4:0.776;HMGB1:0.757).Multivariate analysis identified SF3B4≥1.45,HMGB1≥0.93,poor differentiation,larger tumor size,deeper invasion,lymph node metastasis,and advanced clinical tumor-node-metastasis staging as independent factors contributing to one-year mortality(^(a)P<0.05).Survival analysis indicated that patients with elevated SF3B4 and HMGB1 levels had a shorter median survival(25.74±5.46 months)compared to those with lower expression levels(33.29±6.71 months,log-rank=10.534,^(a)P<0.05).CONCLUSION Elevated SF3B4 and HMGB1 expression in gastric cancer tissue is significantly associated with tumor aggressiveness,worse prognosis,and reduced survival.These biomarkers may offer clinical value in stratifying patients by risk and in forecasting outcomes.Their combined assessment improves predictive accuracy for poor prognosis and may serve as a more effective tool than individual evaluation.
文摘目的:探讨敲低小鼠胚胎腭突间充质细胞(mEPMCs)初级纤毛(PC)重要组件驱动蛋白家族成员3B(KIF3B)基因对细胞自噬水平的作用,并阐明其作用机制。方法:收集体外培养孕龄14.5 d C57BL/6J小鼠的mEPMCs,根据是否敲低KIF3B基因和使用平滑因子受体激动剂(SAG)激活音猬因子(Shh)信号通路及其下游共受体Smo,分为对照组(给予生理盐水)、空载病毒转染细胞组(sh-NC组)(给予慢病毒转染)、敲低KIF3B组(sh-KIF3B组)(给予KIF3B基因敲低)、敲低KIF3B加入SAG组(sh-KIF3B+SAG组)(给予KIF3B基因敲低后加入SAG),每组5只大鼠。透射电镜观察各组mEPMCs中自噬小体/自噬溶酶体形态表现及数量,Western blotting法检测各组mEPMCs中自噬相关蛋白苄氯素1(Beclin-1)和p62及Shh信号通路中Shh和Smo蛋白表达水平。结果:透射电镜观察,与对照组比较,sh-KIF3B组mEPMCs中自噬小体/自噬溶酶体数量明显增加(P<0.05);与sh-KIF3B组比较,sh-KIF3B+SAG组mEPMCs中自噬小体/自噬溶酶体数量明显减少(P<0.05)。Western blotting法检测,与对照组比较,sh-KIF3B组mEPMCs中Beclin-1蛋白表达水平明显升高(P<0.05),KIF3B、p62、Shh和Smo蛋白表达水平均明显降低(P<0.01);与sh-KIF3B组比较,sh-KIF3B+SAG组mEPMCs中Shh、Smo和p62蛋白表达水平明显升高(P<0.01),Beclin-1蛋白表达水平明显降低(P<0.01)。结论:敲低KIF3B基因可促进mEPMCs自噬,其机制可能与其抑制Shh信号通路有关。
