Detection of infrequent and multiple K-ras mutations in human tumors and tumor-adjacent tissues

A sensitive method was developed and applied to examine the distribution of K-ras gene mutations in histologically differing areas of lung tissues obtained from lung cancer patients. This method, which combines polymerase chain reaction (PCR), mutation allele enrichment (MAE), and denaturing gradient gel electrophoresis (DGGE), allows detection of one K-ras mutant allele present in 10(4) to 10(5) wild-type alleles. It was applied to analyze mutations in codon 12 of the K-ras gene in 43 tissue sites microdissected from paraffin-embedded sections obtained from 8 archival cases of lung cancer, all previously shown to have codon 12 K-ras mutations by direct sequencing. In four cases, mutations were detected only in the tumor, while in the other four cases, the same mutations were also found in tissues adjacent to tumors, using the MAE + DGGE method. No mutations were detected among normal-appearing cells in areas distant from the tumors in any of the cases studied. These findings demonstrate that K-ras mutations can be detected at low frequencies in normal-appearing cells from tissues adjacent to the tumor in some lung cancer cases. In addition, this approach also allowed detection of multiple mutations in colorectal tissues obtained from colorectal cancer patients. Thus, the MAE + DGGE method may be applicable to study of K-ras mutations in premalignant or morphologically suspicious lesions in bronchial mucosa or other types of human cancer.     

新型木建筑材料——交叉层积木介绍及其连接的试验研究

交叉层积木作为一种新型木建筑材料已经在北美地区推广开来,然而在我国还没有得到引进和推广.本文介绍了这种新型材料的性能特点,总结其生产工艺、设计方法和研究进展,重点对交叉层积木七种半刚性连接的抗震性能进行试验研究.研究表明:紧固件全部被拔出的失效模式为理想的延性破坏模式,滞回曲线表现出高度非线性、刚度退化、强度退化以及捏拢现象.通过试验数据分析,给出了在延性和承载能力两方面最佳的连接.