F213I突变型高雪氏病分子伴侣治疗方法的研究

目的 研究高雪氏病新的分子治疗方法。方法 用β-葡萄糖脑昔脂酶(β-glucocerebrosidase，β-Glc)(EC3．2．1．45)的底物类似物(glucocerebroside analogue，GCA)作为分子伴侣，处理体外培养的不同突变型的高雪氏病患者皮肤成纤维细胞，采用荧光酶学手段测定β-Glc活性；Western印迹杂交技术分析该酶蛋白表达的量；细胞双染确定β-Glc在细胞内的定位；薄层层析法分析葡萄糖脑苷脂的降解情况。结果 该酶的底物类似物GCA可以提高体外培养的F213I突变型患者成纤维细胞内β-Glc活性；增加该酶蛋白的表达量；促进该酶蛋白向溶酶体内转运，加速底物葡萄糖脑苷脂(glucocerebroside，GlcCer)的降解。结论低分子量的GCA作为一种分子伴侣可能为F213I突变型高雪氏病的治疗开辟一条新的途径。     

Analysis of dynamic cerebral autoregulation using an ARX model based on arterial blood pressure and middle cerebral artery velocity simulation

The study aimed to model the cerebrovascular system, using a linear ARX model based on data simulated by a comprehensive physiological model, and to assess the range of applicability of linear parametric models. Arterial blood pressure (ABP) and middle cerebral arterial blood flow velocity (MCAV) were measured from 11 subjects non-invasively, following step changes in ABP, using the thigh cuff technique. By optimising parameters associated with autoregulation, using a non-linear optimisation technique, the physiological model showed a good performance (r=0.83±0.14) in fitting MCAV. An additional five sets of measured ABP of length 236±154 s were acquired from a subject at rest. These were normalised and rescaled to coefficients of variation (CV=SD/mean) of 2% and 10% for model comparisons. Randomly generated Gaussian noise with standard deviation (SD) from 1% to 5% was added to both ABP and physiologically simulated MCAV (SMCAV), with ‘normal’ and ‘impaired’ cerebral autoregulation, to simulate the real measurement conditions. ABP and SMCAV were fitted by ARX modelling, and cerebral autoregulation was quantified by a 5 s recovery percentage R5% of the step responses of the ARX models. The study suggests that cerebral autoregulation can be assessed by computing the R5% of the step response of an ARX model of appropriate order, even when measurement noise is considerable.     

两株鼠抗人GL50单克隆抗体的制备及其生物学特性的初步研究

目的　研制鼠抗人GL5 0分子单克隆抗体并对其生物学特性进行初步研究。方法　以天然高表达人GL5 0分子的Daudi细胞为免疫原 ,人GL5 0 L92 9转基因细胞为目的单克隆抗体(McAb)的筛选细胞 ,采用B淋巴细胞杂交瘤技术 ,获得分泌特异性鼠抗人GL5 0分子McAb的杂交瘤细胞株 ;免疫荧光标记和流式细胞术分析McAb识别GL5 0的表达 ;Westernblot检测抗体对特异性细胞膜蛋白的识别 ;台盼蓝 (Trypanblue)染色细胞计数法和MTT法检测McAb对Daudi细胞体外生长的抑制作用 ;3 H TdR法检测抗体对GL5 0 L转基因细胞介导的活化T细胞体外增殖的效应。结果　成功制备 2株分泌特异性抗人GL5 0抗体的杂交瘤细胞株 12B11和 11C4 ;两者皆为IgG2a亚型 ;腹水效价皆在 1∶10 0 0以上 ;12B11、11C4特异性地识别GL5 0分子。 11C4McAb可以明显抑制Daudi细胞在体外的生长 ,并可抑制GL5 0 L转基因细胞介导的活化T淋巴细胞的体外增殖效应。结论　成功获得了 2株特异性鼠抗人GL5 0抗体 ,其中 11C4McAb具有诱导表达GL5 0分子的B淋巴瘤细胞Daudi的体外生长抑制作用 ;在T淋巴细胞体外增殖反应中发挥了重要的调节作用。     

Structural Reasons for Restricted Backbone Motion in Poly(n‐alkyl methacrylates): Degree of Polymerization,Tacticity and Side‐Chain Length

Summary: The complex dynamics of poly(n‐alkyl methacrylates) is studied by advanced ¹³C NMR spectroscopy as well as mechanical and dielectric relaxation. Extended backbone conformations are identified as the molecular units involved in structural relaxation. From the variation in the degree of polymerization and a comparison with the presence of stereoregular sequences in the sample, the length of the extended units is determined to involve about five, at most ten monomeric units. Syndiotactic and isotactic sequences behave similarly. These findings are indicative of locally structured polymer melts.

     

Determining appropriate models for joint control using surface electrical stimulation of soleus in spinal cord injury

The mechanical impedance of the ankle joint during electrical stimulation of the soleus is studied by applying constant-velocity 10° angular perturbations to the ankle and measuring the resultant torque. Both neurologically intact subjects and spinal cord injured subjects are tested. Lumped, piecewise linear models are developed to predict the torque from the measured displacement and acceleration signals. The commonly used second-order mass-spring-dashpot model fails to predict the changes in torque that occur following imposed movements. A fiveelement, directionally-dependent piecewise linear model is much better at predicting the measured responses for velocities up to 50° s⁻¹. Numerical least squared error indentification techniques are used to estimate the model parameters for three neurologically intact and three spinal cord injured subjects. The average error between the model’s response and the measured response across all subjects is 10·9%. There is some evidence that a velocity-dependent non-linear model could produce better results than the directionally-dependent piecewise linear model.     

The Origin and Evolution of Human T-Cell Lymphotropic Virus Types I and II

Studies on human T-cell lymphotropic virus types I (HTLV-I) and II (HTLV-II) are briefly reviewed from the viewpoint of molecular evolution, with special reference to the evolutionary rate and evolutionary relationships among these viruses. In particular, it appears that, in contrast to the low level of variability of HTLV-I among different isolates, individual isolates form quasispecies structures. Elucidating the mechanisms connecting these two phenomena will be one of the future problems in the study of the molecular evolution of HTLV-I and HTLV-II.     

Dynamic relationship between EMG and torque at the human ankle: Variation with contraction level and modulation

Stochastic system identification techniques were used to determine the dynamic relationship between the electromyogram (EMG) and torque in the ankle muscles of normal human subjects. EMG and torque were recorded while subjects modulated ankle torque by tracking a computer-generated stochastic waveform. Nonparametric impulse response functions (IRFs) relating EMG to ankle torque were computed and parameterised by determining the parameters of the second-order system which provided the best least-squares fit. Two sets of experiments were carried out. In the first, the mean level of torque was varied from 5 per cent of the maximum voluntary contraction (MVC) to 30 per cent MVC while the depth of modulation was held constant at ±5 per cent of MVC. In the second series of experiments the mean torque was held constant at 25 per cent MVC while the depth of modulation was varied from ±2.5 per cent to ±25 per cent. The major findings were: (1) A second-order, low-pass filter provided a good quasilinear model of the EMG/force dynamics under all conditions; (2) The model parameters depended only weakly on the mean level of torque; (3) In contrast, the model parameters depended strongly on the amplitude with which the contraction was modulated; the natural frequency increased significantly with the depth of modulation.     

人Nanog基因的克隆及其在COS-7L细胞中的表达

目的 :克隆人Nanog基因 ,构建其真核表达载体 ,并观察其在哺乳动物细胞COS 7L中的表达。方法 :利用HE2 93细胞的人基因组DNA为模板 ,以LA PCR技术 ,扩增Nango的基因 ,定向克隆到带Flag标签的pCMV载体中 ,测序后 ,挑选序列正确的真核表达质粒pFlag Nanog转染COS 7L细胞。用抗Flag标签的抗体 ,进行Westernblot和间接免疫荧光染色法检测Nango蛋白的表达。结果 :从人基因组DNA中克隆到序列正确的Nanog全长编码序列。所构建的Nanog质粒在COS 7L细胞中获得高效表达。结论 :人Nanog基因的克隆、真核表达载体的构建及在COS 7L中的表达均获得成功 ,为进一步研究其功能 ,尤其是探讨其在神经干细胞中的作用奠定了基础。     

肺癌靶向多肽荧光分子探针的研制及其应用

目的:选择能与整合素α3受体结合的小分子多肽cNGQGEQc-L作为靶向载体，将羧基荧光素（FAM）与cNGQGEQc-L连接构建荧光分子探针，通过荧光成像探讨荧光多肽分子探针用于肺腺癌显像的可行性。方法:利用倒置荧光显微镜观察FAM-cNGQGEQc-L与肺腺癌A549细胞结合部位，流式细胞仪检测荧光多肽与A549细胞的竞争抑制实验，观察FAM-cNGQGEQc-L随浓度的增加与肺腺癌A549细胞结合能力的变化情况。通过小动物活体成像仪，观察荧光多肽在荷瘤裸鼠体内的生物分布特点。结果:倒置荧光显微镜显示荧光多肽cNGQGEQc-L能与A549细胞结合，结合部位在细胞膜和细胞质中。流式细胞仪测试结果证明荧光多肽与A549细胞的结合具有特异性和饱和性，当FAM-cNGQGEQc-L浓度为0.125 mmol/L时，荧光多肽与A549细胞的结合趋近饱和。荷瘤裸鼠活体成像显示移植瘤能够摄取荧光多肽，且荧光多肽通过泌尿系统和胆道系统排泄。结论:体外、体内荧光实验结果显示，荧光多肽分子探针FAM-cNGQGEQc-L可与肺腺癌A549细胞、肺癌移植瘤结合，能够特异性靶向肺腺癌。