Preparation and properties of ^(117m)Sn(Ⅳ)-TTHMP in in-vitro and in small animals

In this work,TTHMP was synthesized and labelled with ^(117m)Sn.The preparation conditions,stability and lipophilicity of ^(117m)Sn(Ⅳ)-TTHMP were investigated.Biodistribution of the complex in rabbits and mice was studied. It was found that the quantity of TTHMP and pH value of the preparation solution had vital effects on the labeling yield of ^(117m)Sn(Ⅳ)-TTHMP.It was also found that ^(117)Sn(Ⅳ)-TTHMP was hydrophilic and stable at room tempera- ture and 37℃in open air.^(117m)Sn(Ⅳ)-TTHMP showed unexpectedly high bone uptake and bone-to-blood ratio in the animals.This made it potentially useful as an reagent for skeletal scintigraphy and radiotherapy of bone tumors.

应用Bac-to-Bac系统高效表达人类错配修复基因hMLH1及其错义突变体

目的利用Bac-to-Bac杆状病毒表达系统在Sf9昆虫细胞中实现人类错配修复基因h MLH1及其错义突变体的高效表达。方法分别用Bam HI和Not I双酶切构建于pc DNA载体中的全长野生型h MLH1基因及T117M和V384D错义突变型DNA片段,回收后定向克隆至p Fast Bac1转座载体中,经酶切鉴定后转化DH10Bac感受态细胞,PCR方法筛选阳性菌落,提取重组Bacmid,反复感染Sf9昆虫细胞,用SDS-PAGE和Western blot方法鉴定目的蛋白的表达。结果提取转染72h后的Sf9细胞总蛋白,用7%的SDS-聚丙烯酰胺凝胶电泳,可见分子量为85k Da清晰的蛋白条带,经Western blot方法证实该条带为h MLH1基因的表达产物。结论利用Bac-to-Bac杆状病毒表达系统在Sf9昆虫细胞中大量表达出h MLH1野生型和T117M及V384D错义突变体蛋白,为进一步开展体外错配修复实验奠定了良好的基础。

多种^(117)Sn^m(^(113)Sn)-配体体系的形成条件及其与BSA的结合

以117Snm(113Sn)示踪法研究了Sn—DTPMP、Sn—DTPA、Sn—EDTMP、Sn—HEDT、MP、Sn—TTHMP体系的形成条件,同时研究了不同标记体系与牛血清白蛋白(BSA)的结合性质。研究表明,除DTPA外,其他配体在较高浓度下均能与Sn4+形成络合,但配体浓度较低时锡极易水解,用生理盐水稀释和增加体系pH均加剧水解。研究同时显示,水解锡标记体系与BSA有很强的结合,未水解锡标记体系与BSA的结合率远低于水解锡标记体系。