The protein connector enhancer of kinase suppressor of Ras 2(CNKSR2),present in both the postsynaptic density and cytoplasm of neurons,is a scaffolding protein with several protein-binding domains.Variants of the CNKS...The protein connector enhancer of kinase suppressor of Ras 2(CNKSR2),present in both the postsynaptic density and cytoplasm of neurons,is a scaffolding protein with several protein-binding domains.Variants of the CNKSR2 gene have been implicated in neurodevelopmental disorders,particularly intellectual disability,although the precise mechanism involved has not yet been fully understood.Research has demonstrated that CNKSR2 plays a role in facilitating the localization of postsynaptic density protein complexes to the membrane,thereby influencing synaptic signaling and the morphogenesis of dendritic spines.However,the function of CNKSR2 in the cytoplasm remains to be elucidated.In this study,we used immunoprecipitation and high-resolution liquid chromatography-mass spectrometry to identify the interactors of CNKSR2.Through a combination of bioinformatic analysis and cytological experiments,we found that the CNKSR2 interactors were significantly enriched in the proteome of the centrosome.We also showed that CNKSR2 interacted with the microtubule protein DYNC1H1 and with the centrosome marker CEP290.Subsequent colocalization analysis confirmed the centrosomal localization of CNKSR2.When we downregulated CNKSR2 expression in mouse neuroblastoma cells(Neuro 2A),we observed significant changes in the expression of numerous centrosomal genes.This manipulation also affected centrosome-related functions,including cell size and shape,cell proliferation,and motility.Furthermore,we found that CNKSR2 interactors were highly enriched in de novo variants associated with intellectual disability and autism spectrum disorder.Our findings establish a connection between CNKSR2 and the centrosome,and offer new insights into the underlying mechanisms of neurodevelopmental disorders.展开更多
BACKGROUND Trastuzumab constitutes the fundamental component of initial therapy for patients with advanced human epidermal growth factor receptor 2(HER-2)-positive gastric cancer(GC).However,the efficacy of this treat...BACKGROUND Trastuzumab constitutes the fundamental component of initial therapy for patients with advanced human epidermal growth factor receptor 2(HER-2)-positive gastric cancer(GC).However,the efficacy of this treatment is hindered by substantial challenges associated with both primary and acquired drug resistance.While S-phase kinase associated protein 2(Skp2)overexpression has been implicated in the malignant progression of GC,its role in regulating trastuzumab resistance in this context remains uncertain.Despite the numerous studies investigating Skp2 inhibitors among small molecule compounds and natural products,there has been a lack of successful commercialization of drugs specifically targeting Skp2.AIM To discover a Skp2 blocker among currently available medications and develop a therapeutic strategy for HER2-positive GC patients who have experienced progression following trastuzumab-based treatment.METHODS Skp2 exogenous overexpression plasmids and small interfering RNA vectors were utilized to investigate the correlation between Skp2 expression and trastuzumab resistance in GC cells.Q-PCR,western blot,and immunohistochemical analyses were conducted to evaluate the regulatory effect of thioridazine on Skp2 expression.A cell counting kit-8 assay,flow cytometry,a amplex red glucose/glucose oxidase assay kit,and a lactate assay kit were utilized to measure the proliferation,apoptosis,and glycolytic activity of GC cells in vitro.A xenograft model established with human GC in nude mice was used to assess thioridazine's effectiveness in vivo.RESULTS The expression of Skp2 exhibited a negative correlation with the sensitivity of HER2-positive GC cells to trastuzumab.Thioridazine demonstrated the ability to directly bind to Skp2,resulting in a reduction in Skp2 expression at both the transcriptional and translational levels.Moreover,thioridazine effectively inhibited cell proliferation,exhibited antiapoptotic properties,and decreased the glucose uptake rate and lactate production by suppressing Skp2/protein kinase B/mammalian target of rapamycin/glucose transporter type 1 signaling pathways.The combination of thioridazine with either trastuzumab or lapatinib exhibited a more pronounced anticancer effect in vivo,surpassing the efficacy of either monotherapy.CONCLUSION Thioridazine demonstrates promising outcomes in preclinical GC models and offers a novel therapeutic approach for addressing trastuzumab resistance,particularly when used in conjunction with lapatinib.This compound has potential benefits for patients with Skp2-proficient tumors.展开更多
The present study was designed to determine microtubule-associated protein-2 and synaptophysin expression in the hippocampal CA3 region in a rat model of middle cerebral artery occlusion. The rats were treated with ac...The present study was designed to determine microtubule-associated protein-2 and synaptophysin expression in the hippocampal CA3 region in a rat model of middle cerebral artery occlusion. The rats were treated with acupuncture at Baihui （GV 20）, Qubin （GB 7）, and Qianding （GV 21） points, in addition to exercise training. Results were compared with rats undergoing exercise training only. The Y-maze method and immunohistochemistry revealed decreased error frequency of passing through Y-maze, as well as significantly increased microtubule-associated protein-2 and synaptophysin expression, in the acupuncture with exercise training group compared with the model and exercise training groups after 5 weeks. Microtubule-associated protein-2 and synaptophysin expressions negatively correlated with error frequency of passing through the Y-maze. These results suggested that acupuncture combined with exercise training improved learning and memory functions in a rat model of cerebral infarction. The mechanisms of action were hypothesized to be associated with dendritic or synaptic plasticity in the ipsilateral hippocampal CA3 region.展开更多
Following granulocyte colony-stimulating factor （G-CSF） treatment,the growth of processes in cul-tured rat retinal ganglion cells （RGCs） in vitro,expression of growth associated protein 43,and expression of microt...Following granulocyte colony-stimulating factor （G-CSF） treatment,the growth of processes in cul-tured rat retinal ganglion cells （RGCs） in vitro,expression of growth associated protein 43,and expression of microtubule-associated protein 2 mRNA expression were significantly increased.In contrast,RhoA/Rock protein content was significantly reduced by G-CSF treatment.These results indicate that G-CSF promotes the growth of processes in RGCs and increases the expression of growth-associated protein 43 and microtubule-associated protein 2 mRNA by inhibiting the RhoA/Rock pathway,thereby benefiting axonal repair in RGCs exposed to hypoxia.展开更多
目的探讨胃衡汤调控核因子E2相关因子2(Nrf2)-Kelch样环氧氯丙烷相关蛋白(Keap)信号通路抑制老年胃癌前病变(Precancerous lesions of gastric cancer,PLGC)患者氧化应激反应的作用。方法选取2022年3月-2023年3月期间于南京中医药大学...目的探讨胃衡汤调控核因子E2相关因子2(Nrf2)-Kelch样环氧氯丙烷相关蛋白(Keap)信号通路抑制老年胃癌前病变(Precancerous lesions of gastric cancer,PLGC)患者氧化应激反应的作用。方法选取2022年3月-2023年3月期间于南京中医药大学第二附属医院消化腔镜中心和脾胃病科就诊的PLGC老年患者60例,按随机数字表法分为基础治疗组与胃衡汤治疗组,每组各30例。另选择同期常规胃镜体检的慢性浅表性胃炎患者30例为对照组。对照组仅入组接受研究相关指标检测,不予治疗。基础治疗组患者给予雷贝拉唑钠肠溶胶囊,胃衡汤组在基础治疗组上加用胃衡汤治疗,均治疗6个月。观察比较两组PLGC患者临床疗效、安全性,治疗前后中医证候评分、胃镜病理分级及Nrf2-Keap通路相关蛋白检测。结果治疗后胃衡汤治疗组中医证候疗效总有效率93.33%(28/30)明显高于基础治疗组76.67%(23/30),差异有统计学意义(P<0.05)。治疗后胃衡汤治疗组病理组织学疗效总有效率83.33%(25/30)明显高于基础治疗组63.33%(19/30),差异有统计学意义(P<0.05)。治疗后两组患者中医证候各项积分及总积分均较治疗前降低,差异有统计学意义(P<0.05);且胃衡汤治疗组中医证候各项积分及总积分均明显低于基础治疗组,差异有统计学意义(P<0.05)。治疗后两组患者胃黏膜组织学各项积分及总分均较治疗前降低,差异有统计学意义(P<0.05);且胃衡汤治疗组胃黏膜组织学各项积分及总分均明显低于基础治疗组,差异有统计学意义(P<0.05)。治疗后两组患者Nrf2和SOD表达高于治疗前,Keap1和MDA表达较治疗前降低,差异有统计学意义(P<0.05);且胃衡汤治疗组Nrf2和SOD表达明显高于基础治疗组,Keap1和MDA表达明显低于基础治疗组,差异有统计学意义(P<0.05)。治疗期间,两组患者治疗前后肝肾功能、三大常规及心电图检查均未发现异常。结论胃衡汤治疗老年PLGC能够改善临床症状,抑制甚至逆转病理学改变,提高治疗效果,其机制与调节Nrf2-Keap信号通路而抑制氧化应激反应有关。展开更多
目的分析儿童抗接触蛋白相关蛋白-2(contactin-associated protein 2,Caspr2)抗体相关自身免疫性脑炎的临床特点、诊疗过程及预后情况,以期提高儿科医生对此病的认识。方法回顾性收集2019年6月至2022年6月山东大学附属儿童医院收治的抗C...目的分析儿童抗接触蛋白相关蛋白-2(contactin-associated protein 2,Caspr2)抗体相关自身免疫性脑炎的临床特点、诊疗过程及预后情况,以期提高儿科医生对此病的认识。方法回顾性收集2019年6月至2022年6月山东大学附属儿童医院收治的抗Caspr2抗体相关脑炎患儿4例,分析其临床资料的特征并进行随访。结果男女患儿各2例,年龄为1.3~7.0岁。4例均呈亚急性发病,首发症状表现为发热、惊厥发作4例,表现为自主神经功能障碍2例、头痛1例、精神行为异常1例;主要临床表现为意识障碍3例,言语障碍、运动障碍各2例,睡眠增多、反复头痛各1例。4例患儿头颅MRI检查和脑电图检查结果均存在异常。3例脑脊液Caspr2抗体阳性,4例血清Caspr2抗体阳性(其中1例复发时呈阳性)。3例行糖皮质激素联合免疫球蛋白治疗,1例行支持治疗,出院时病情均好转。出院后18~28个月进行随访,1例出现复发,4例改良Rankin评分0~2分。结论儿童抗Caspr2抗体相关脑炎临床表现不典型,常表现为意识障碍、发热、精神行为异常、睡眠障碍、癫痫、运动障碍、自主神经症状等。患儿对免疫治疗有较好的反应,早期诊断、早期治疗患儿预后良好。展开更多
基金supported by the National Nature Science Foundation of China,No.32101020(to JL)the Natural Science Foundation of Shandong Province,Nos.ZR2020MC071(to JL),ZR2023MH327(to HZ)+1 种基金the Integrated Project of Major Research Plan of National Natural Science Foundation of China,No.92249303(to PL)the Natural Science Foundation of Qingdao,No.23-2-1-193-zyyd-jch(to HZ)。
文摘The protein connector enhancer of kinase suppressor of Ras 2(CNKSR2),present in both the postsynaptic density and cytoplasm of neurons,is a scaffolding protein with several protein-binding domains.Variants of the CNKSR2 gene have been implicated in neurodevelopmental disorders,particularly intellectual disability,although the precise mechanism involved has not yet been fully understood.Research has demonstrated that CNKSR2 plays a role in facilitating the localization of postsynaptic density protein complexes to the membrane,thereby influencing synaptic signaling and the morphogenesis of dendritic spines.However,the function of CNKSR2 in the cytoplasm remains to be elucidated.In this study,we used immunoprecipitation and high-resolution liquid chromatography-mass spectrometry to identify the interactors of CNKSR2.Through a combination of bioinformatic analysis and cytological experiments,we found that the CNKSR2 interactors were significantly enriched in the proteome of the centrosome.We also showed that CNKSR2 interacted with the microtubule protein DYNC1H1 and with the centrosome marker CEP290.Subsequent colocalization analysis confirmed the centrosomal localization of CNKSR2.When we downregulated CNKSR2 expression in mouse neuroblastoma cells(Neuro 2A),we observed significant changes in the expression of numerous centrosomal genes.This manipulation also affected centrosome-related functions,including cell size and shape,cell proliferation,and motility.Furthermore,we found that CNKSR2 interactors were highly enriched in de novo variants associated with intellectual disability and autism spectrum disorder.Our findings establish a connection between CNKSR2 and the centrosome,and offer new insights into the underlying mechanisms of neurodevelopmental disorders.
基金Supported by Youth Fund of National Natural Science Foundation of China,No.81803575,No.31902287Kaifeng Science and Technology Development Plan Project,No.2203008+2 种基金Key Specialized Research and Promotion Project of Henan Province in 2023,No.232102311205Henan Medical Science and Technology Research Program Project,No.LHGJ20210801College Students Innovation and Entrepreneurship Training Program of Henan University,No.20231022007.
文摘BACKGROUND Trastuzumab constitutes the fundamental component of initial therapy for patients with advanced human epidermal growth factor receptor 2(HER-2)-positive gastric cancer(GC).However,the efficacy of this treatment is hindered by substantial challenges associated with both primary and acquired drug resistance.While S-phase kinase associated protein 2(Skp2)overexpression has been implicated in the malignant progression of GC,its role in regulating trastuzumab resistance in this context remains uncertain.Despite the numerous studies investigating Skp2 inhibitors among small molecule compounds and natural products,there has been a lack of successful commercialization of drugs specifically targeting Skp2.AIM To discover a Skp2 blocker among currently available medications and develop a therapeutic strategy for HER2-positive GC patients who have experienced progression following trastuzumab-based treatment.METHODS Skp2 exogenous overexpression plasmids and small interfering RNA vectors were utilized to investigate the correlation between Skp2 expression and trastuzumab resistance in GC cells.Q-PCR,western blot,and immunohistochemical analyses were conducted to evaluate the regulatory effect of thioridazine on Skp2 expression.A cell counting kit-8 assay,flow cytometry,a amplex red glucose/glucose oxidase assay kit,and a lactate assay kit were utilized to measure the proliferation,apoptosis,and glycolytic activity of GC cells in vitro.A xenograft model established with human GC in nude mice was used to assess thioridazine's effectiveness in vivo.RESULTS The expression of Skp2 exhibited a negative correlation with the sensitivity of HER2-positive GC cells to trastuzumab.Thioridazine demonstrated the ability to directly bind to Skp2,resulting in a reduction in Skp2 expression at both the transcriptional and translational levels.Moreover,thioridazine effectively inhibited cell proliferation,exhibited antiapoptotic properties,and decreased the glucose uptake rate and lactate production by suppressing Skp2/protein kinase B/mammalian target of rapamycin/glucose transporter type 1 signaling pathways.The combination of thioridazine with either trastuzumab or lapatinib exhibited a more pronounced anticancer effect in vivo,surpassing the efficacy of either monotherapy.CONCLUSION Thioridazine demonstrates promising outcomes in preclinical GC models and offers a novel therapeutic approach for addressing trastuzumab resistance,particularly when used in conjunction with lapatinib.This compound has potential benefits for patients with Skp2-proficient tumors.
文摘The present study was designed to determine microtubule-associated protein-2 and synaptophysin expression in the hippocampal CA3 region in a rat model of middle cerebral artery occlusion. The rats were treated with acupuncture at Baihui （GV 20）, Qubin （GB 7）, and Qianding （GV 21） points, in addition to exercise training. Results were compared with rats undergoing exercise training only. The Y-maze method and immunohistochemistry revealed decreased error frequency of passing through Y-maze, as well as significantly increased microtubule-associated protein-2 and synaptophysin expression, in the acupuncture with exercise training group compared with the model and exercise training groups after 5 weeks. Microtubule-associated protein-2 and synaptophysin expressions negatively correlated with error frequency of passing through the Y-maze. These results suggested that acupuncture combined with exercise training improved learning and memory functions in a rat model of cerebral infarction. The mechanisms of action were hypothesized to be associated with dendritic or synaptic plasticity in the ipsilateral hippocampal CA3 region.
文摘Following granulocyte colony-stimulating factor （G-CSF） treatment,the growth of processes in cul-tured rat retinal ganglion cells （RGCs） in vitro,expression of growth associated protein 43,and expression of microtubule-associated protein 2 mRNA expression were significantly increased.In contrast,RhoA/Rock protein content was significantly reduced by G-CSF treatment.These results indicate that G-CSF promotes the growth of processes in RGCs and increases the expression of growth-associated protein 43 and microtubule-associated protein 2 mRNA by inhibiting the RhoA/Rock pathway,thereby benefiting axonal repair in RGCs exposed to hypoxia.
文摘目的探讨胃衡汤调控核因子E2相关因子2(Nrf2)-Kelch样环氧氯丙烷相关蛋白(Keap)信号通路抑制老年胃癌前病变(Precancerous lesions of gastric cancer,PLGC)患者氧化应激反应的作用。方法选取2022年3月-2023年3月期间于南京中医药大学第二附属医院消化腔镜中心和脾胃病科就诊的PLGC老年患者60例,按随机数字表法分为基础治疗组与胃衡汤治疗组,每组各30例。另选择同期常规胃镜体检的慢性浅表性胃炎患者30例为对照组。对照组仅入组接受研究相关指标检测,不予治疗。基础治疗组患者给予雷贝拉唑钠肠溶胶囊,胃衡汤组在基础治疗组上加用胃衡汤治疗,均治疗6个月。观察比较两组PLGC患者临床疗效、安全性,治疗前后中医证候评分、胃镜病理分级及Nrf2-Keap通路相关蛋白检测。结果治疗后胃衡汤治疗组中医证候疗效总有效率93.33%(28/30)明显高于基础治疗组76.67%(23/30),差异有统计学意义(P<0.05)。治疗后胃衡汤治疗组病理组织学疗效总有效率83.33%(25/30)明显高于基础治疗组63.33%(19/30),差异有统计学意义(P<0.05)。治疗后两组患者中医证候各项积分及总积分均较治疗前降低,差异有统计学意义(P<0.05);且胃衡汤治疗组中医证候各项积分及总积分均明显低于基础治疗组,差异有统计学意义(P<0.05)。治疗后两组患者胃黏膜组织学各项积分及总分均较治疗前降低,差异有统计学意义(P<0.05);且胃衡汤治疗组胃黏膜组织学各项积分及总分均明显低于基础治疗组,差异有统计学意义(P<0.05)。治疗后两组患者Nrf2和SOD表达高于治疗前,Keap1和MDA表达较治疗前降低,差异有统计学意义(P<0.05);且胃衡汤治疗组Nrf2和SOD表达明显高于基础治疗组,Keap1和MDA表达明显低于基础治疗组,差异有统计学意义(P<0.05)。治疗期间,两组患者治疗前后肝肾功能、三大常规及心电图检查均未发现异常。结论胃衡汤治疗老年PLGC能够改善临床症状,抑制甚至逆转病理学改变,提高治疗效果,其机制与调节Nrf2-Keap信号通路而抑制氧化应激反应有关。
文摘目的分析儿童抗接触蛋白相关蛋白-2(contactin-associated protein 2,Caspr2)抗体相关自身免疫性脑炎的临床特点、诊疗过程及预后情况,以期提高儿科医生对此病的认识。方法回顾性收集2019年6月至2022年6月山东大学附属儿童医院收治的抗Caspr2抗体相关脑炎患儿4例,分析其临床资料的特征并进行随访。结果男女患儿各2例,年龄为1.3~7.0岁。4例均呈亚急性发病,首发症状表现为发热、惊厥发作4例,表现为自主神经功能障碍2例、头痛1例、精神行为异常1例;主要临床表现为意识障碍3例,言语障碍、运动障碍各2例,睡眠增多、反复头痛各1例。4例患儿头颅MRI检查和脑电图检查结果均存在异常。3例脑脊液Caspr2抗体阳性,4例血清Caspr2抗体阳性(其中1例复发时呈阳性)。3例行糖皮质激素联合免疫球蛋白治疗,1例行支持治疗,出院时病情均好转。出院后18~28个月进行随访,1例出现复发,4例改良Rankin评分0~2分。结论儿童抗Caspr2抗体相关脑炎临床表现不典型,常表现为意识障碍、发热、精神行为异常、睡眠障碍、癫痫、运动障碍、自主神经症状等。患儿对免疫治疗有较好的反应,早期诊断、早期治疗患儿预后良好。
