Mechanism Study of the Promoting Action of Elongated Needle Acupuncture on Functional Repair in Rats with Acute Spinal Cord Injury

目的 观察芒针对急性脊髓损伤炎症反应和神经细胞凋亡的影响,研究PI3K/Akt和MAPK/ERK信号转导途径是否参与芒针的神经保护作用,探讨芒针促脊髓损伤修复的具体作用机制.方法 将150只成年雄性SD大鼠随机分为A组、B组、C组、D组和E组,每组30只.采用改良Allen＇s法制作大鼠脊髓中度损伤模型,A组为假手术组,不予以损伤脊髓及芒针治疗;B组造模后不予以芒针治疗;C组造模后采用芒针治疗;D组造模前0.5 h鞘内注射LY294002,造模后采用芒针治疗;E组造模前0.5 h鞘内注射PD98059,造模后采用芒针治疗.分别采用BBB评分检测大鼠的自发活动;ELISA检测炎性因子TNF-α 、IL-6、IL-1β、NF-κB的含量;TUNEL检测细胞凋亡的程度;免疫组化检测Bcl-2和Bax阳性细胞水平;Western印迹分析p-Akt和p-ERK在脊髓组织的表达,RT-PCR分析Cyt-C和Caspase-3在体内的表达.相对的下行Akt和ERK信号通路通过LY294002和PD98059特异性抑制剂处理分析在体内的抑制作用.结果 炎症反应和PI3K/Akt和MAPK/ERK信号通路抑制的神经元凋亡参与了脊髓损伤大鼠模型的损害,芒针介导的神经保护作用与Bax蛋白阳性神经元数目的减少及Bcl-2蛋白阳性神经元数量的增加有关.芒针治疗能改善大鼠的运动功能,PI3K/Akt和MAPK/ERK信号通路中p-Akt和p-ERK的激活,通过下调Bax蛋白和Bcl-2表达上调,抑制了线粒体凋亡途径关键因子Cyt-C的表达.TUNEL法检测并抑制激活神经元凋亡的Caspase-3的级联.PI3K/Akt和MAPK/ERK信号通路特异性抑制剂LY294002和PD98059的应用抑制了p-Akt和p-ERK的表达.结论 芒针促脊髓损伤修复的神经保护作用可能与炎症反应和PI3K/Akt和MAPK/ERK信号通路的激活、通过下调Bax蛋白和Bcl-2表达上调以及抑制线粒体途径诱导的凋亡有关.Objective To observe the effect of elongated needle acupuncture on inflammatory reactions and apoptosis in acute spinal cord injury, investigate if PI3K/Akt and MAPK/ERK signal transduction pathways are involved in the neuroprotective effect of elongated needle acupuncture and explore the promoting action of elongated needle acupuncture on spinal cord injury repair.Method One hundred and fifty adult male SD rats were randomized into groups A, B, C, D and E, 30 each. A model of moderate spinal cord injury was made by modified Allen＇s method. Group A received a sham operation without spinal cord injury and no elongated needle acupuncture. Group B did not receive elongated needle acupuncture after model making. Group C received elongated needle acupuncture after model making. Group D received an intrathecal injection of LY294002 at 0.5 hour before model making and elongated needle acupuncture after model making. Group E received an intrathecal injection of PD98059 at 0.5 hour before model making and elongated needle acupuncture after model making. Rat spontaneous activity was examined using the BBB rating. Inflammatory cytokines TNF-α, IL-6, IL-1β and NF-κB contents were measured by ELISA. The degree of apoptosis was determined by TUNEL. Bcl-2- and Bax-positive cell levels were measured by an immunohistochemical method. Spinal p-Akt and p-ERK expressions were determined by Western blot. In vivo expressions of Cyt-C and Caspase-3 were determined by RT-PCR. The in vivo inhibitory effect on downstream Akt and ERK signaling pathways was investigated using specific inhibitors LY294002 and PD98059.Result Inflammatory reactions and neuronal apoptosis due to the inhibition of PI3K/Akt and MAPK/ERK signal pathways were involved in the damage in a rat model of spinal cord injury. The neuroprotective effect of elongated needle acupuncture was related to a decrease in the number of Bax protein-positive neurons and an increase in the number of Bcl-2 protein-positive neurons. Elongated needle acupuncture treatment improved r浙江省中医药科研计划项目（2008CB067

Maroteaux-Lamy综合征的ARSB基因分析及新突变的致病性鉴定

【目的】对7家拟诊为Maroteaux_Lamy综合征(MPSⅥ)的患儿及其父母进行ARSB基因的突变检测和新突变的致病性鉴定,以揭示其分子发病机制,为将来的产前/植入前基因诊断等创造前提条件。【方法】在临床初诊及GAG尿检和MPS酶检的基础上,抽取患儿及其父母EDTA抗凝血,进行ARSB基因的PCR扩增和Sanger测序。对所发现的新突变,在经HGMD、1000G和ExAC等数据库核实排查后,首先用SWISS-MODEL软件分析、比对突变蛋白和正常蛋白的空间构象,然后用Clustal X软件分析跨物种氨基酸的保守性,用PROVEAN、SIFT、PolyPhen-2软件预测新突变的致病性,最后用ACMG标准对新突变的致病性进行综合分析鉴定。【结果】1)7个家系先证者的基因检测结果分别为:No1:c.574T> C,p.C192R纯合错义突变;No2:c.160G> A/p.D54N(来自其母)和c.1197C> G/p.F399L(来自其父)的复合杂合子;No3:仅检出c.1072G> A/p.V358M和IVS5 as(-27)A>C变异,但酶检和临床表型符合Ⅵ型;No4:为c.281C> T,p.S94L(新突变,来自其母)和IVS5 as(-27)A> C(来自其父)的复合杂合子;No5:为c.1197 C> G,p.F399L纯合错义突变;No6:为c.1197 C> G,p.F399L(来自其母)和c.1379 C> T,p.S460F(新突变,来自其父)的复合杂合子;No7:为c.499 G> A,p.G167R(来自其父)和c.1325C>T,p.T442M(来自其母)的复合杂合子。2)新突变鉴定结果:对正常ARSB酶蛋白和p.S94L突变酶蛋白的空间构象的预测比对结果显示,两者有明显区别;跨物种保守性分析结果显示,p.94突变点所在氨基酸(S)在物种进化过程中具有高度保守性;PROVEAN、SIFT和PolyPhen-2软件对p.S94L预测结果分别为:Deleterious、Damaging和Probably damaging。用上述方法对p.S460F的预测结果及ACMG的综合分析结果也显示该突变可能是致病性的。【结论】1)家系4的p.S94L和家系6的p.S460F新突变可能都是新的致病性突变,有可能都是引起患儿发病的内在原因之一。2)家系1,2,5,6,7可以确诊为MPS Ⅵ型,其基因型和表现型具有显著的相关性,家系3虽经酶检确诊,临床症状和尿检结果也都与MPS Ⅵ型符合,但却未能在DNA水平查到明确的突变类型,其表现型与基因型的相关性还有待进一步证实。国家自然科学基金(30772069);;闽粤合作科研基金(71010025和71020010

新型稠环电子受体的界面修饰对钙钛矿太阳能电池性能的影响

钙钛矿太阳能电池(perovskite solar cells, PSCs)因具有能量转换效率(power conversion efficiency, PCE)高、成本低、易于大面积制造等优点而被科学家们广泛关注.氧化物电子传输层的合理界面设计及修饰对提高器件的PCE和工作长期稳定性有着十分重要的意义.因此,本文采用一种含有烷基噻吩基侧链的稠环电子受体材料3,9-二(2-亚甲基-(3-(1,1-二氰甲烯基)-茚酮))-5,5,11,11-四(5-己基噻吩)-二噻吩并[2,3-d:2′,3′-d′]-s-引达省[1,2-b:5,6-b′]二噻吩(3,9-bis(2-methylene-(3-(1,1-dicyanomethylene)-indanone))-5,5,11,11-tetrakis(5-hexylthienyl)-dithieno[2,3-d:2′,3′-d′]-s-indaceno[1,2-b:5,6-b′]dithiophene, ITIC-Th)修饰TiO2电子传输层,制备高效稳定的平面结PSCs.研究结果显示,ITIC-Th的界面修饰改善了TiO2薄膜的形貌、接触角等性质,促进了钙钛矿晶粒的高质量生长,大幅度减少了器件表界面的电荷复合,明显提升了光生载流子的抽取率和输运效率,使经ITIC-Th界面修饰的PSC的PCE从未经界面修饰的15.43%显著提高到18.91%.与此同时,器件稳定性的研究结果显示,在室温和湿度为30%的条件下,经ITICTh界面修饰的PSC的PCE在放置约1000 h后依然保持原来的90%,明显高于未经界面修饰的PSC.研究结果对PSC光伏性能的提升具有重要的实际应用价值和学术意义.国家自然科学基金(61605164);;\n陕西省重点科技创新团队计划(2016KCT-28);;\n陕西省重点研发计划(2017ZDXM-GY-046);;\n福建省科技厅高校产学合作项目(2016H6023)资

Effects of electro-acupuncture on neuronal apoptosis and associative function in rats with spinal cord injury

目的 :探讨电针对大鼠急性脊髓损伤后膀胱功能改善的影响及作用机制。方法 :取健康成年雄性SPf级Sd大鼠60只,体重220~250 g,适应性饲养1周后,将大鼠按照随机数字表法随机分为假手术组、模型组、电针组和电针对照组,各15只。假手术组不予任何刺激,模型组、电针组和电针对照组大鼠采用改良AllEnS法制作脊髓损伤中度损伤Sd大鼠模型,模型组不予治疗,电针组给予秩边与水道穴电针治疗,电针对照组给予秩边与水道穴旁开0.5寸电针治疗,频率2/100 Hz,电流1 M A,刺激15 MIn,电针左右隔次交替,每日1次,共7次;分别于术后1、7 d观察大鼠残余尿量、排尿量的变化;术后7 d处死大鼠取伤段脊髓观察各组大鼠凋亡情况,检测bCl-2、bAX、bAd含量的变化。结果:造模后3组大鼠均出现不同程度的膀胱功能障碍。术后7 d,电针组、电针对照组残余尿量较术后1 d明显降低(P<0.001),且电针组与电针对照组比较差异有统计学意义(P<0.01);电针组、电针对照组较模型组在术后7 d排尿量增加,且电针组与电针对照组比较差异有统计学意义(P<0.001);TunEl发现电针可以抑制脊髓神经细胞的凋亡,电针组、电针对照组与模型组相比在术后7 d脊髓神经细胞凋亡率显著增加(P<0.01,P<0.05),且电针组与电针对照组比较差异有统计学意义(P<0.05);与模型组相比,电针组、电针对照组bAX、bAd的阳性表达率降低(P<0.01,P<0.05),而bCl-2的阳性表达率升高(P<0.01);且电针组与电针对照组比较差异有统计学意义(P<0.05)。结论 :电针能明显促进急性脊髓损伤的修复,其机制可能为通过增加bCl-2、抑制bAX、bAd的表达,从而抑制脊髓神经元细胞的凋亡发生作用的。Objective:To explore the effect of electro acupuncture to improve the bladder function after acute spinal cord injury in rats and its possible mechanism.Methods:Sixty healthy adult male SD rats of SPF grade,with body weight of 220 to250 g,one week after feeding adaptation,were randomly divided into sham operation group,model group,electro acupuncture group,electro acupuncture control group with 15 rats in each group.Sham operation group underwent no stimulation,and the moderate damage model of spinal cord injury were made in other three groups according to modified Allens method.The model group were not treated,electro acupuncture group were treated with electro acupuncture on Zhibianxue and Shuidaoxue,and electro acupuncture control group were treated with electro acupuncture on 0.5 inch next to Zhibianxue and Shuidaoxue.The frequency of 2 / 100 Hz,current of 1 m A,stimulation time of 15 min,once a day,left and right alternately stimulate every time,for a total of 7 times.The changes of residual urine volume and urine output in rats at the 1st and the 7th days after operation were observed.And 7 d later,the rats were sacrificed and the injured spinal cord were taken out to observe the apoptosis,and to detect the changes of Bcl-2,Bax,Bad content.Results:After modeling,the rats of three groups showed different bladder dysfunction.In electro acupuncture group and electro acupuncture control group,the residual urine volume of the 7th day after operation was significant lower than the 1st day after operation(P<0.001),and there was statistically significant difference on the 7th day after operation between two groups(P<0.001).Compared with model group,the urine output of electro acupuncture group and electro acupuncture control group was significantly increased on the 7th day after operation,and there was significant difference between electro acupuncture group and electro acupuncture control group(P<0.001).Electro acupuncture can inhibit apoptosis of spinal cord neurons by TUNEL detection.Postoperative at 7 d,the rate of nerve cell apoptosis in electro acupuncture group and electro acupuncture control group was significant increased than model group(P<0.01,P<0.05),and there was significant difference between electro acupuncture group and electro acupuncture control group(P<0.005).Compared with model group,the positive expression rate of Bax,Bad decreased(P<0.01,P<0.05),and Bcl-2 increased(P<0.01)in electro acupuncture group and electro acupuncture control group,there was significant difference between electro acupuncture group and electro acupuncture control group(P<0.01).Conclusion:Electro acupuncture can obviously promote the repair of acute spinal cord injury,its mechanism may be through increasing Bcl-2,inhibiting the expression of Bax,Bad,which inhibits the apoptosis of spinal cord neurons.浙江省中医药科研计划项目(编号:2008CB067)~

Maroteaux-Lamy综合征的ARSB基因分析及新突变的致病性鉴定

【目的】对7家拟诊为Maroteaux_Lamy综合征（MPSⅥ）的患儿及其父母进行ARSB基因的突变检测 和新突变的致病性鉴定，以揭示其分子发病机制，为将来的产前/植入前基因诊断等创造前提条件。【方法】在 临床初诊及GAG尿检和MPS酶检的基础上，抽取患儿及其父母EDTA抗凝血，进行ARSB基因的PCR扩增和 Sanger测序。对所发现的新突变，在经HGMD、1000G和ExAC等数据库核实排查后，首先用SWISS-MODEL软件 分析、比对突变蛋白和正常蛋白的空间构象，然后用ClustalX软件分析跨物种氨基酸的保守性，用PROVEAN、 SIFT、PolyPhen-2软件预测新突变的致病性，最后用ACMG标准对新突变的致病性进行综合分析鉴定。【结果】 1） 7个家系先证者的基因检测结果分别为：No1： c.574T>C，p.C192R纯合错义突变；No2： c.160G>A/p.D54N（来 自其母）和c.1197C>G/p.F399L（来自其父）的复合杂合子；No3：仅检出c.1072G>A/p.V358M和IVS5as（-27）A> C变异，但酶检和临床表型符合VI型；No4：为c.281C>T，p.S94L（新突变，来自其母）和IVS5as（-27）A>C（来自 其父）的复合杂合子；No5：为c.1197C>G，p.F399L纯合错义突变；No6：为c.1197C>G，p.F399L（来自其母）和 c.1379C>T，p.S460F（新突变，来自其父）的复合杂合子；No7：为c.499G>A，p.G167R（来自其父）和c.1325C> T，p.T442M（来自其母）的复合杂合子。2）新突变鉴定结果：对正常ARSB酶蛋白和p.S94L突变酶蛋白的空间构 象的预测比对结果显示，两者有明显区别；跨物种保守性分析结果显示， p.94突变点所在氨基酸（S）在物种进化 过程中具有高度保守性；PROVEAN、SIFT和PolyPhen-2软件对p.S94L预测结果分别为：Deleterious、Damaging和 Probablydamaging。用上述方法对p.S460F的预测结果及ACMG的综合分析结果也显示该突变可能是致病性 的。【结论】1）家系4的p.S94L和家系6的p.S460F新突变可能都是新的致病性突变，有可能都是引起患儿发病 的内在原因之一。2）家系1， 2， 5， 6， 7可以确诊为MPSVI型，其基因型和表现型具有显著的相关性，家系3虽经 酶检确诊，临床症状和尿检结果也都与MPSVI型符合，但却未能在DNA水平查到明确的突变类型，其表现型与 基因型的相关性还有待进一步证实

局部开孔对矩形钢板抗剪承载能力影响研究

由于优越的抗剪性能，钢板被广泛的应用于建筑结构以及船舶海洋工程中。在实际工程中，由于使用功能的需要或者锈蚀等原因，钢板开孔的现象屡见不鲜。开孔会减小钢板的有效抗剪面积进而降低抗剪承载能力。针对矩形孔和圆形孔两种开孔类型对钢板抗剪承载力的影响展开了研究。通过试验研究了开孔位置，开孔率对钢板抗剪承载力的影响。结果表明：开孔位置对钢板抗剪承载力有显著影响；抗剪承载力的降低程度不能简单通过开孔率进行评估。在此基础上，通过理论和试验验证了数值模型的可靠性，进而通过参数化分析研究了两种开孔形状不同位置和高宽比对钢板抗剪承载力的影响规律，提出了采用折减系数反映抗剪承载力的降低程度，并提出了折减系数的理论计算公式。通过对比结果表明，所提出的理论计算公式可以准确反映开孔位置对钢板抗剪承载力的影响，可为开孔钢板抗剪承载力的准确评估奠定基础

地震分频解释技术在三角洲体系中的应用以东营凹陷沙三中亚段东营三角洲为例

本次研究在基于多元标定和井震对比的前提下,对东营凹陷沙三中亚段东营三角洲进行了高精度层序地层格架的建立,识别并追踪了目的层段关键的三级层序界面和三角洲进积体内部不同期次的进积单元界面,并总结了各个界面的沉积相、岩相和地震相组合特征.在此基础上选取典型地震剖面进行了地震分频处理,认为不同频段地震资料其所揭示的地质信息是存在差异的.分频地震处理的结果显示,低频资料能够清晰地显示三级层序界面和三角洲进积体内部结构差异,可用于划分内部的进积单元;高频资料能够显示出进积单元内部的高频旋回.利用高频地震资料所进行的高频旋回划分,与钻井岩性、测井曲线响应和测井曲线小波变换的结果具有很好的对应程度