The problems of movement apparatus in children, youth and even adolescents aren’t connected with “a weakness of muscles” but with a shortening of muscles, tendons, and capsules which in orthopaedic literature is ca...The problems of movement apparatus in children, youth and even adolescents aren’t connected with “a weakness of muscles” but with a shortening of muscles, tendons, and capsules which in orthopaedic literature is called “contracture” [1] [2] [3] [4]. The older way of thinking about the problem was based on the conviction that “weak muscles” cause and make problems;we, however, present on many examples that “restriction of movements” doing by shortening of soft tissues makes contracture and incorrect position of joints, body parts, the serious and frequent clinical problems.展开更多
目的探讨黑皮质素受体激动剂(MT-Ⅱ)改善SH3和多个锚蛋白重复结构域3(Shank3)基因缺陷孤独症模型鼠社交缺陷的作用机制。方法利用Shank3干扰慢病毒和空载慢病毒注射至仔鼠右侧脑室构建Shank3模型鼠和空载鼠各18只,Shank3组随机分为Shank...目的探讨黑皮质素受体激动剂(MT-Ⅱ)改善SH3和多个锚蛋白重复结构域3(Shank3)基因缺陷孤独症模型鼠社交缺陷的作用机制。方法利用Shank3干扰慢病毒和空载慢病毒注射至仔鼠右侧脑室构建Shank3模型鼠和空载鼠各18只,Shank3组随机分为Shank3+Sal(Sh3-Sal)组9只和Shank3+MT-Ⅱ(Sh3-MT-Ⅱ)组9只,空载组随机分为空载+Sal(V-Sal)组9只和空载+MT-Ⅱ(V-MT-Ⅱ)组9只。V-MT-Ⅱ组和Sh3-MT-Ⅱ组于第28天腹腔注射3.3 m L/kg MT-Ⅱ,V-Sal组和Sh3-Sal组腹腔注射3.3 m L/kg 0.9%氯化钠溶液,通过旷场实验、理毛实验、三箱社交实验及Morris水迷宫实验评估其行为学改变;采用逆转录聚合酶链式反应(RT-PCR)和蛋白免疫印迹(Western blot)检测下丘脑催产素(OXT)、催产素受体(OXTR)及黑皮质素受体4(MC4R)的mRNA与蛋白表达水平。结果行为学结果显示,三箱社交实验中与陌生鼠1相比,Sh3-Sal组未表现出社交差异(P>0.05),而MT-Ⅱ干预后,Sh3-MT-Ⅱ组与陌生鼠2的社交时间显著增加,差异有统计学意义(P<0.01)。Morris水迷宫实验中与V-Sal组相比,Sh3-Sal组表现出显著的学习记忆障碍(P<0.05),而MTⅡ干预后,Sh3-MT-Ⅱ组学习记忆能力明显提高,差异有统计学意义(P<0.01)。旷场实验和理毛实验结果显示,与V-Sal组相比,Sh3-Sal组周边停留时间及理毛时间均显著增加,差异有统计学意义(P<0.01);MT-Ⅱ干预后,旷场中心停留时间及理毛行为与Sh3-Sal组无显著差异(P>0.05)。RT-PCR检测显示,与Sh3-Sal组相比,Sh3-MT-Ⅱ组OXT、OXTR和MC4R mRNA表达水平明显升高,差异有统计学意义(P<0.05);Western blot法检测显示,与Sh3-Sal组相比,Sh3-MT-Ⅱ组大鼠下丘脑OXT蛋白表达水平明显升高(P<0.05),与V-Sal组相比,Sh3-Sal组和Sh3-MT-Ⅱ组大鼠下丘脑SHANK3蛋白表达水平明显降低(P<0.05,P<0.01),而OXTR及MC4R蛋白表达水平无显著差异(P>0.05)。结论黑皮质素受体激动剂MT-Ⅱ可能通过激活下丘脑OXT系统改善Shank3缺陷孤独症模型鼠的社交障碍,提示靶向OXT/MC4R通路或为孤独症社交缺陷的潜在干预策略。展开更多
Children with autism often exhibit abnormalities in body weight,but the underlying mechanism remains unclear.SH3 and multiple ankyrin repeat domains protein 3(SHANK3),a scaffold protein of the postsynaptic density,has...Children with autism often exhibit abnormalities in body weight,but the underlying mechanism remains unclear.SH3 and multiple ankyrin repeat domains protein 3(SHANK3),a scaffold protein of the postsynaptic density,has been reported to be associated with autism.This study aimed to investigate whether and how SHANK3 influences body weight in the hypothalamic neuronal regulation of energy homeostasis.Adeno-associated viruses 9(AAV9)carrying CMV-Cre and Agrp-Cre were stereotactically injected to restore SHANK3 expression in the arcuate nucleus(ARC)and agouti-related peptide(AgRP)neurons,respectively.Agrp-Cre mice were injected with AAV9-p38αflox/flox to overexpress p38α.Activated p38αwas generated by mutating both D176A and F327S in p38α.Inactivated p38αwas constructed by mutating both T180A and Y182F in p38α.Metabolic analysis,immunoblotting,histological analysis,the glucose tolerance test,the insulin tolerance test,and body fat mass analysis were applied to investigate the underlying mechanisms by which SHANK3 regulates body weight.We reveal that SHANK3 regulates body weight via the p38αsignaling pathway in the AgRP neurons of the hypothalamus.Shank3 knockout(Shank3−/−)mice exhibit resistance to diet-induced obesity.Shank3 re-expression in the ARC or AgRP neurons increases body weight in Shank3 knock-in mice with an inverted allele(SKO).Overexpression or activation of p38αin AgRP neurons elicits resistance to diet-induced obesity.Inactivated p38αin AgRP neurons abolished the resistance to diet-induced obesity due to SHANK3 deficiency.Our findings suggest that the SHANK3-p38αsiganling pathway in AgRP neurons regulates body weight balance in autism,revealing a promising therapeutic target for obesity in children with autism.展开更多
Autism Spectrum Disorder(ASD)is marked by early-onset neurodevelopmental anomalies,yet the tem-poral dynamics of genetic contributions to these processes remain insufficiently understood.This study aimed to elu-cidate...Autism Spectrum Disorder(ASD)is marked by early-onset neurodevelopmental anomalies,yet the tem-poral dynamics of genetic contributions to these processes remain insufficiently understood.This study aimed to elu-cidate the role of the Shank3 gene,known to be associated with monogenic causes of autism,in early developmental processes to inform the timing and mechanisms for poten-tial interventions for ASD.Utilizing the Shank3B knockout(KO)mouse model,we examined Shank3 expression and its impact on neuronal maturation through Golgi staining for dendritic morphology and electrophysiological recordings to measure synaptic function in the anterior cingulate cortex(ACC)across different postnatal stages.Our longitudinal analysis revealed that,while Shank3B KO mice displayed normal neuronal morphology at one week postnatal,signifi-cant impairments in dendritic growth and synaptic activity emerged by two to three weeks.These findings highlight the critical developmental window during which Shank3 is essential for neuronal and synaptic maturation in the ACC.展开更多
目的观察5肽类似物165对老年性痴呆(Alzhei mer disease,AD)模型大鼠学习记忆能力及突触后致密区蛋白95(postsynaptic density95,PSD95)和骨架蛋白Shank1表达的影响。方法将45只大鼠随机分为正常对照组、模型组和5肽类似物165治疗组,模...目的观察5肽类似物165对老年性痴呆(Alzhei mer disease,AD)模型大鼠学习记忆能力及突触后致密区蛋白95(postsynaptic density95,PSD95)和骨架蛋白Shank1表达的影响。方法将45只大鼠随机分为正常对照组、模型组和5肽类似物165治疗组,模型组和治疗组大鼠按体重3mg/kg行双侧侧脑室链脲佐菌素(streptozotocin,STZ)注射,第3天重复注射建立AD模型,对照组以人工脑脊液代替STZ。术后21d治疗组按体重0.34mg/(kg.d)给予APP5肽类似物165灌胃干预,其余两组以蒸馏水代替。3周后应用Morris水迷宫、免疫组织化学和Western blotting方法检测大鼠的学习记忆能力及PSD95和Shank1的表达。结果5肽类似物165治疗组大鼠的平均游泳时间较模型组明显缩短(P<0.01),且海马PSD95和Shank1阳性神经细胞数及PSD95和Shank1蛋白表达较模型组明显增加(P<0.05)。结论5肽类似物165可显著提高大鼠学习记忆能力,增加大鼠海马PSD95和Shank1表达,表明其对突触功能和可塑性具改善作用。展开更多
文摘The problems of movement apparatus in children, youth and even adolescents aren’t connected with “a weakness of muscles” but with a shortening of muscles, tendons, and capsules which in orthopaedic literature is called “contracture” [1] [2] [3] [4]. The older way of thinking about the problem was based on the conviction that “weak muscles” cause and make problems;we, however, present on many examples that “restriction of movements” doing by shortening of soft tissues makes contracture and incorrect position of joints, body parts, the serious and frequent clinical problems.
文摘目的探讨黑皮质素受体激动剂(MT-Ⅱ)改善SH3和多个锚蛋白重复结构域3(Shank3)基因缺陷孤独症模型鼠社交缺陷的作用机制。方法利用Shank3干扰慢病毒和空载慢病毒注射至仔鼠右侧脑室构建Shank3模型鼠和空载鼠各18只,Shank3组随机分为Shank3+Sal(Sh3-Sal)组9只和Shank3+MT-Ⅱ(Sh3-MT-Ⅱ)组9只,空载组随机分为空载+Sal(V-Sal)组9只和空载+MT-Ⅱ(V-MT-Ⅱ)组9只。V-MT-Ⅱ组和Sh3-MT-Ⅱ组于第28天腹腔注射3.3 m L/kg MT-Ⅱ,V-Sal组和Sh3-Sal组腹腔注射3.3 m L/kg 0.9%氯化钠溶液,通过旷场实验、理毛实验、三箱社交实验及Morris水迷宫实验评估其行为学改变;采用逆转录聚合酶链式反应(RT-PCR)和蛋白免疫印迹(Western blot)检测下丘脑催产素(OXT)、催产素受体(OXTR)及黑皮质素受体4(MC4R)的mRNA与蛋白表达水平。结果行为学结果显示,三箱社交实验中与陌生鼠1相比,Sh3-Sal组未表现出社交差异(P>0.05),而MT-Ⅱ干预后,Sh3-MT-Ⅱ组与陌生鼠2的社交时间显著增加,差异有统计学意义(P<0.01)。Morris水迷宫实验中与V-Sal组相比,Sh3-Sal组表现出显著的学习记忆障碍(P<0.05),而MTⅡ干预后,Sh3-MT-Ⅱ组学习记忆能力明显提高,差异有统计学意义(P<0.01)。旷场实验和理毛实验结果显示,与V-Sal组相比,Sh3-Sal组周边停留时间及理毛时间均显著增加,差异有统计学意义(P<0.01);MT-Ⅱ干预后,旷场中心停留时间及理毛行为与Sh3-Sal组无显著差异(P>0.05)。RT-PCR检测显示,与Sh3-Sal组相比,Sh3-MT-Ⅱ组OXT、OXTR和MC4R mRNA表达水平明显升高,差异有统计学意义(P<0.05);Western blot法检测显示,与Sh3-Sal组相比,Sh3-MT-Ⅱ组大鼠下丘脑OXT蛋白表达水平明显升高(P<0.05),与V-Sal组相比,Sh3-Sal组和Sh3-MT-Ⅱ组大鼠下丘脑SHANK3蛋白表达水平明显降低(P<0.05,P<0.01),而OXTR及MC4R蛋白表达水平无显著差异(P>0.05)。结论黑皮质素受体激动剂MT-Ⅱ可能通过激活下丘脑OXT系统改善Shank3缺陷孤独症模型鼠的社交障碍,提示靶向OXT/MC4R通路或为孤独症社交缺陷的潜在干预策略。
基金supported by the Noncommunicable Chronic Diseases-National Science and Technology Major Project(2023ZD0506800).
文摘Children with autism often exhibit abnormalities in body weight,but the underlying mechanism remains unclear.SH3 and multiple ankyrin repeat domains protein 3(SHANK3),a scaffold protein of the postsynaptic density,has been reported to be associated with autism.This study aimed to investigate whether and how SHANK3 influences body weight in the hypothalamic neuronal regulation of energy homeostasis.Adeno-associated viruses 9(AAV9)carrying CMV-Cre and Agrp-Cre were stereotactically injected to restore SHANK3 expression in the arcuate nucleus(ARC)and agouti-related peptide(AgRP)neurons,respectively.Agrp-Cre mice were injected with AAV9-p38αflox/flox to overexpress p38α.Activated p38αwas generated by mutating both D176A and F327S in p38α.Inactivated p38αwas constructed by mutating both T180A and Y182F in p38α.Metabolic analysis,immunoblotting,histological analysis,the glucose tolerance test,the insulin tolerance test,and body fat mass analysis were applied to investigate the underlying mechanisms by which SHANK3 regulates body weight.We reveal that SHANK3 regulates body weight via the p38αsignaling pathway in the AgRP neurons of the hypothalamus.Shank3 knockout(Shank3−/−)mice exhibit resistance to diet-induced obesity.Shank3 re-expression in the ARC or AgRP neurons increases body weight in Shank3 knock-in mice with an inverted allele(SKO).Overexpression or activation of p38αin AgRP neurons elicits resistance to diet-induced obesity.Inactivated p38αin AgRP neurons abolished the resistance to diet-induced obesity due to SHANK3 deficiency.Our findings suggest that the SHANK3-p38αsiganling pathway in AgRP neurons regulates body weight balance in autism,revealing a promising therapeutic target for obesity in children with autism.
基金supported by the Natural Science Foundation of Anhui Province,China(No.2408085MH189)the Scientific Research Program of University in Anhui Province,China(No.2024AH051017,KJ2021A0202)+4 种基金the Training Program for Young and Middle-aged Teachers in University of Anhui Province,China(No.YQYB2024031)the Clinical Science Research Project of the First Affiliated Hospital of Anhui University of Chinese Medicine(No.2021yfylc14)the Biomedical Basic Applied Science and Technology Public Service Platform of Large-scale Instruments and Equipment Open Fund of Anhui Province,China(No.2023-04)the Students'Innovation and Entrepreneurship Training Program of Anhui Province,China(No.S202410366078)the Scientific Research Platform and Base Upgrading Plan of Anhui Medical University(No.2021xkjT048)。
基金supported by the Natural Science Foundation of China(32394032,82201699,and 82221001)the Natural Science Foundation of Zhejiang Province(LTGD24H250001)+1 种基金the Kay R&D Program of Shaanxi Province(2023-YBSF-093),the Young Talent Fund of University Association for Science and Technology in Shaanxi(20220306)the Joint Founding Project of Innovation Research Institute,Xijing Hospital(LHJJ24JH02).
文摘Autism Spectrum Disorder(ASD)is marked by early-onset neurodevelopmental anomalies,yet the tem-poral dynamics of genetic contributions to these processes remain insufficiently understood.This study aimed to elu-cidate the role of the Shank3 gene,known to be associated with monogenic causes of autism,in early developmental processes to inform the timing and mechanisms for poten-tial interventions for ASD.Utilizing the Shank3B knockout(KO)mouse model,we examined Shank3 expression and its impact on neuronal maturation through Golgi staining for dendritic morphology and electrophysiological recordings to measure synaptic function in the anterior cingulate cortex(ACC)across different postnatal stages.Our longitudinal analysis revealed that,while Shank3B KO mice displayed normal neuronal morphology at one week postnatal,signifi-cant impairments in dendritic growth and synaptic activity emerged by two to three weeks.These findings highlight the critical developmental window during which Shank3 is essential for neuronal and synaptic maturation in the ACC.
文摘目的观察5肽类似物165对老年性痴呆(Alzhei mer disease,AD)模型大鼠学习记忆能力及突触后致密区蛋白95(postsynaptic density95,PSD95)和骨架蛋白Shank1表达的影响。方法将45只大鼠随机分为正常对照组、模型组和5肽类似物165治疗组,模型组和治疗组大鼠按体重3mg/kg行双侧侧脑室链脲佐菌素(streptozotocin,STZ)注射,第3天重复注射建立AD模型,对照组以人工脑脊液代替STZ。术后21d治疗组按体重0.34mg/(kg.d)给予APP5肽类似物165灌胃干预,其余两组以蒸馏水代替。3周后应用Morris水迷宫、免疫组织化学和Western blotting方法检测大鼠的学习记忆能力及PSD95和Shank1的表达。结果5肽类似物165治疗组大鼠的平均游泳时间较模型组明显缩短(P<0.01),且海马PSD95和Shank1阳性神经细胞数及PSD95和Shank1蛋白表达较模型组明显增加(P<0.05)。结论5肽类似物165可显著提高大鼠学习记忆能力,增加大鼠海马PSD95和Shank1表达,表明其对突触功能和可塑性具改善作用。
