Na+ channel expression and neuronal function in the Na+/H+ exchanger 1 null mutant mouse

Mice lacking Na(+)/H(+) exchanger 1 (NHE1) suffer from recurrent seizures and die early postnatally. Although the mechanisms for seizures are not well established, our previous electrophysiological work has shown that neuronal excitability and Na(+) current density are increased in hippocampal CA1 neurons of these mutant mice. However, it is unknown whether this increased density is related to altered expression or functional regulation of Na(+) channels. In this work, we asked three questions: is the increased excitability limited to CA1 neurons, is the increased Na(+) current density related to an increased Na(+) channel expression, and, if so, which Na(+) channel subtype(s) is upregulated? Using neurophysiological, autoradiographic, and immunoblotting techniques, we showed that both CA1 and cortical neurons have an increase in membrane excitability and Na(+) current density; Na(+) channel density is selectively upregulated in the hippocampus and cortex (P < 0.05); and Na(+) channel subtype I is significantly increased in the hippocampus and Na(+) channel subtype II is increased in the cortex. Our results demonstrate that mice lacking NHE1 upregulate their Na(+) channel expression in the hippocampal and cortical regions selectively; this leads to an increase in Na(+) current density and membrane excitability. We speculate that neuronal overexcitability due to Na(+) channel upregulation in the hippocampus and cortex forms the basis of epileptic seizures in NHE1 mutant mice.     

The myogenic response in isolated rat cerebrovascular arteries: vessel model

We develop an integrated model of isolated rat arterial resistance vessel (RV), which can simulate its major property of myogenic response. The vascular smooth muscle cell is an important component of the wall of this vessel, and serves as a vasomotor organ providing the active tension generation that underlies the myogenic response of the wall to stretch. In the previous study, we focused on the development of a smooth muscle cell model that can mimic the strain-sensing and force-generating features of the myogenic mechanism. In the current model, we embed this cell model in a larger vessel wall configuration, and couple the time course of cellular contractile activation to macroscopic changes in vessel diameter. The integrated model is used to mimic published pressure-vessel diameter data obtained from isolated RVs that are mounted in a hydraulic test apparatus. The model provides biophysically based insights into the myogenic mechanism as it responds to changes in transmural pressure, in the presence and absence of Ca2+ blockers applied to the bathing fluid.It mimics measured data very well and provides a model that is able to link events at subcellular level to macroscopic changes in vessel diameter. The model initiates a mechanistic approach to investigate myogenic response, which has not been taken previously by any other models.     

不同结合状态硫酸软骨素多糖链对体外二维三维培养背根节神经突起生长影响研究

目的构建鸡胚背根节细胞3D生长模型,观察CSPG对不同浓度的琼脂糖水溶胶培养基对鸡胚背根节神经突起生长的影响。方法①利用多肽缩合剂1’1羰基二咪唑介导CS-B长链与琼脂糖凝胶共价结合。②分别配制0.5%、0.75%、1%、1.25%、1.5%的SeaPrep琼脂糖凝胶溶液和共价结合了CS-B多糖长链的琼脂糖凝胶溶液。取孵育9~10天后鸡胚背根节加入上述凝胶溶液加培养液培养。③培养24小时之后在24 h、48 h7、2 h、96 h进行观察拍照。观察各时间点神经突起生长情况。结果①鸡胚背根节神经突起在1.25%的琼脂糖凝胶溶液中开始呈3D生长,在小于1%的琼脂糖溶液中只能2D生长。②在同一时间点,呈2D生长的背根节突起和呈3D生长的背根节突起在突起长度方面没有统计学差异(P>0.05)。③3D生长的DRG神经突起在共价结合了CS-B的琼脂糖凝胶中生长长度明显短于普通琼脂糖水凝胶(P<0.05),并且可观察到部分突起长出短而直的成束突起。但是对于呈2D生长的DRG突起长度在两种培养基中却未见差异(P>0.05)。结论鸡胚背根节神经元周围突起生长长度在3D培养中与在2D培养中有没有明显的差异。硫酸软骨素能够有效抑制呈3D生长的神经节突起生长,并且影响到神经突起的生长方式。但是在2D培养基中,未能看到上述的抑制作用。     

核素唾液腺显像的定量分析软件开发与应用

目的开发定量分析唾液腺功能及其图象显示的程序。方法应用Aladdin语言编写ROI模块提取各唾液腺及甲状腺区域的摄取数据及本底,用曲线函数生成各ROI的时间-放射性活度曲线,对曲线数据处理,计算唾液腺摄取、排泌指数等指标,然后把图象、曲线及计算结果显示在一个模板内。结果同一幅输出界面图可显示唾液腺的形态、ROI示意图、唾液腺的摄取与排泌功能数值及其曲线,以标准化的显示格式提供给临床医师。不同操作者处理同一资料的功能指标结果基本一致,软件计算的功能指数与图象显示结果及临床诊断基本相符。结论开发的唾液腺功能定量分析软件,程序运行稳定,结果重复性好。     

十二指肠乳头部肝样腺癌一例

患者男,74岁.以无明显诱因出现黑便伴尿黄、眼黄1个月于2008年10月7日入院.胃镜诊断:十二指肠球部溃疡,降段癌?MRI示十二指肠乳头区占位性病变并肝内外胆管扩张.血癌胚抗原((CEA)1.91μg/L,血CA199 40.76U/ml,肝功能未见明显异常,术前查2次甲胎蛋白(AFP)分别为349及509μg/L.行胰十二指肠切除术.     

应用复合诱导突变分离PCR法检测mtDNA的单核苷酸多态性

目的应用复合诱导突变分离PCR(multiplexed mutagenically separated PCR,MS-PCR)技术、银染分型,建立线粒体DNA(mitochondrial DNA,mtDNA)编码区单核苷酸多态(single nucleotide polymorphism,SNP)分型系统,探讨其应用价值。并调查了成都汉族群体mtDNA编码区4个SNP基因座等位基因频率和单倍型分布情况。方法根据SNP基因座(C12705T、A8701G、G8584A、C10400T)设计两条片段相差4个碱基的等位基因特异性引物和一条公共引物,4个SNP基因座复合扩增,PCR产物经聚丙烯酰胺凝胶电泳、银染显带后确定样本的基因型。结果不同SNP基因座为长度不同的单一谱带,其分型结果与直接测序一致。在成都汉族160名无关个体中,4个SNP基因座C12705T、A8701G、G8584A、C10400T等位基因频率分别为0.3813/0.6187、0.4813/0·5187、0.8250/0.1750、0.4938/0.5062;共检出6种单倍型,单倍型的基因多样性为0.7137。结论建立的MMS-PCR银染分型系统是一种简单、快速、准确、有效的SNP分型方法,对建立mtDNA编码区SNP数据库,研究群体遗传学、进化学和进行法医学个人识别和亲子鉴定有重要意义。     

华南、华北人群HLA-DQB1等位基因多态性

目的从基因水平调查了中国华南、华北地区人群HLA-DQB1等位基因频率，并研究比较两地区人群HLA-DQB1多态性分布。方法采用深圳益生堂生物企业有限公司研制开发的“HLA-DQB1低分辨率分型基因芯片检测试剂盒”，应用聚合酶链反应．序列特异性引物＋序列特异性寡核苷酸探针芯片检测技术，对700名南方地区的中国人和320名北方地区的中国人进行基因分型。结果鉴定了10个HLA-DQB1等位基因，获得了一组准确、科学的统计数据。结论得到了中国华南、华北地区人群HLA-DQB1等位基因频率差异的数据，证明中国人群HLA-DQB1＊02，05，0601，0602，0603的分布南北差异有统计学意义（P〈0．05），为疾病相关性研究、人文科学研究提供了可靠的遗传学数据。     

大白鼠外侧隔核的传入性神经纤维联系—HRP、荧光素法研究

本文应用HRP和荧光素标记法对大白鼠外侧隔核的传入性神经纤维联系进行了研究。实验结果表明,外侧隔核的传入神经纤维来源于端脑海马的CA_(1-4)区、杏仁皮质核、内嗅区皮质、斜角带核、视前内侧核、视前外侧核;丘脑的室周核和带旁核;底丘脑的Forel's H_2区和未定带;下丘脑的前核、外侧核、后核以及乳头体上核;中脑的腹侧被盖区。