[目的]建立一种新的优化的小鼠中脑黑质取材方法,用于检测帕金森病(Parkinson′s disease, PD)小鼠黑质区域的细胞和分子改变。[方法] 2月龄C57BL/6J小鼠随机分为传统组织定位取材组和脑片定位取材组,取黑质组织,通过Western blotting和...[目的]建立一种新的优化的小鼠中脑黑质取材方法,用于检测帕金森病(Parkinson′s disease, PD)小鼠黑质区域的细胞和分子改变。[方法] 2月龄C57BL/6J小鼠随机分为传统组织定位取材组和脑片定位取材组,取黑质组织,通过Western blotting和qPCR方法检测两种取材方法黑质中酪氨酸羟化酶(Tyrosine hydroxylase, TH)的含量。随后通过向小鼠黑质立体定位注射AAV-α-synuclein病毒载体建立PD模型组,脑片定位取材方法获取黑质,应用RNA测序和qPCR检测该取材方法能否有效反应PD相关基因的改变。[结果]脑片定位取材组1~6片脑片中,4号脑片TH蛋白含量显著高于组织定位取材组5.98倍(24.03±1.823 vs 4.018±3.675)及1号、2号、6号脑片(1;3.841±4.333;2.23±1.494);富集3、4、5号脑片提取RNA,其Th基因表达水平高于组织定位取材组2.3倍(2.303±0.4162 vs 1)及1、2、6号脑片富集组4.04倍(2.303±0.4162 vs 0.5746±0.0812)。在PD小鼠模型中富集3、4、5号脑片,转录组测序结果提示,多巴胺神经元相关基因下调,免疫功能相关基因上调,qPCR验证了PD模型黑质中Th和Aldh1a7表达水平显著下降(0.9018±0.138 vs 0.2854±0.09;1.031±0.3262 vs 0.1703±0.07989),CD180和Ddx60表达水平显著增加(1.008±0.1556 vs 4.597±1.586;1.007±0.1417 vs 12.94±2.776)。[结论]相较于组织定位取材方法,脑片定位取材能够更准确的提取到黑质组织,并且能够有效捕捉到多巴胺能神经元的丢失情况以及PD相关基因的改变,对于检测PD动物模型中的细胞和分子变化的精准性具有重要意义。展开更多
Objective To clearly delineate the microanatomy of the upper thoracic sympathetic trunk and its syntopy in order to provide anatomical basis for the selection of precise clinical surgical approaches.Methods Totally 55...Objective To clearly delineate the microanatomy of the upper thoracic sympathetic trunk and its syntopy in order to provide anatomical basis for the selection of precise clinical surgical approaches.Methods Totally 55 sympathetic trunks available in 28 Chinese adult cadavers(18 males and 10 females)were investigated in this study.The thoracic cavity was thoroughly exposed and then sympathetic ganglia and its affiliated rami were observed and measured with microscopic instruments.Results Rami from thoracic nerve 1(T1)intercostal nerve entered the stellate ganglion above the upper border of the second rib in all specimens,The distance between the rami and the upper border of the second rib was(5.1±1.4)mm.The incidence that T2⁃T5 ganglions located at the corresponding intercostal space was 92.7%,85.5%,40.0%and 36.4%,respectively.The rami communicantes were classified into 3 types(type a,b and c)according to the anatomical relationship between the thoracic sympathetic ganglia and the intercostal nerves.The distance from 3 types of rami communicantes to lateral sympathetic trunk at the middle point of relative rib was(10.2±3.4)mm(type a),(9.5±2.6)mm(type b)and(8.4±2.0)mm(type c),respectively.We found that 74.5%superior intercostal artery crossed the neck of the first rib.The distance between the superior intercostal artery and the lateral sympathetic trunk was(3.4±1.3)mm.Conclusion Sympathectomy performed at rib level should be the ideal choice for clinical surgical treatment of palmar or craniofacial hyperhidrosis with less denervation scope to achieve more definite therapeutic effect.展开更多
文摘[目的]建立一种新的优化的小鼠中脑黑质取材方法,用于检测帕金森病(Parkinson′s disease, PD)小鼠黑质区域的细胞和分子改变。[方法] 2月龄C57BL/6J小鼠随机分为传统组织定位取材组和脑片定位取材组,取黑质组织,通过Western blotting和qPCR方法检测两种取材方法黑质中酪氨酸羟化酶(Tyrosine hydroxylase, TH)的含量。随后通过向小鼠黑质立体定位注射AAV-α-synuclein病毒载体建立PD模型组,脑片定位取材方法获取黑质,应用RNA测序和qPCR检测该取材方法能否有效反应PD相关基因的改变。[结果]脑片定位取材组1~6片脑片中,4号脑片TH蛋白含量显著高于组织定位取材组5.98倍(24.03±1.823 vs 4.018±3.675)及1号、2号、6号脑片(1;3.841±4.333;2.23±1.494);富集3、4、5号脑片提取RNA,其Th基因表达水平高于组织定位取材组2.3倍(2.303±0.4162 vs 1)及1、2、6号脑片富集组4.04倍(2.303±0.4162 vs 0.5746±0.0812)。在PD小鼠模型中富集3、4、5号脑片,转录组测序结果提示,多巴胺神经元相关基因下调,免疫功能相关基因上调,qPCR验证了PD模型黑质中Th和Aldh1a7表达水平显著下降(0.9018±0.138 vs 0.2854±0.09;1.031±0.3262 vs 0.1703±0.07989),CD180和Ddx60表达水平显著增加(1.008±0.1556 vs 4.597±1.586;1.007±0.1417 vs 12.94±2.776)。[结论]相较于组织定位取材方法,脑片定位取材能够更准确的提取到黑质组织,并且能够有效捕捉到多巴胺能神经元的丢失情况以及PD相关基因的改变,对于检测PD动物模型中的细胞和分子变化的精准性具有重要意义。
文摘Objective To clearly delineate the microanatomy of the upper thoracic sympathetic trunk and its syntopy in order to provide anatomical basis for the selection of precise clinical surgical approaches.Methods Totally 55 sympathetic trunks available in 28 Chinese adult cadavers(18 males and 10 females)were investigated in this study.The thoracic cavity was thoroughly exposed and then sympathetic ganglia and its affiliated rami were observed and measured with microscopic instruments.Results Rami from thoracic nerve 1(T1)intercostal nerve entered the stellate ganglion above the upper border of the second rib in all specimens,The distance between the rami and the upper border of the second rib was(5.1±1.4)mm.The incidence that T2⁃T5 ganglions located at the corresponding intercostal space was 92.7%,85.5%,40.0%and 36.4%,respectively.The rami communicantes were classified into 3 types(type a,b and c)according to the anatomical relationship between the thoracic sympathetic ganglia and the intercostal nerves.The distance from 3 types of rami communicantes to lateral sympathetic trunk at the middle point of relative rib was(10.2±3.4)mm(type a),(9.5±2.6)mm(type b)and(8.4±2.0)mm(type c),respectively.We found that 74.5%superior intercostal artery crossed the neck of the first rib.The distance between the superior intercostal artery and the lateral sympathetic trunk was(3.4±1.3)mm.Conclusion Sympathectomy performed at rib level should be the ideal choice for clinical surgical treatment of palmar or craniofacial hyperhidrosis with less denervation scope to achieve more definite therapeutic effect.
