应用大规模测序技术和生物信息学研究造血干/祖细胞的基因表达及新基因 的识别和克隆

从新生儿脐血和成人骨髓中分选出造血干/祖细胞(HSC/HPC),构建成cDNA文库,对其进行大规模表达序列标签(EST)测序,通过生物信息学等手段分析基因表达谱,并进行新基因的全长cDNA克隆。在所测的10512条可分析E ST序列中,有9866条来自脐血CD34+|细胞,其中4697条(47.6%)为已知基因,2603条(26.4%)为已知EST,1415条(14.3%)代表未知EST。在已知基因中,8.2%基因与造血相关,22.7%涉及细胞代谢、结构和迁移,13.0%与细胞分裂和防御相关,26.2%与RNA、蛋白质的合成相关,10.6%和细胞信号传递有关。对一些已知和未知的EST,综合测序、生物信息学等方法,进行全长克隆,已获得23个新基因的全长cDNA。 Abstract:Hematopoietic stem/progenitor cells were isolated from umbilical cord blood and adult bone marrow,and subject to cDNA library construction.The gene expression pattern in CD34+ cells and the identification and cloning of novel genes were performed by sequencing ESTs and analyzing them with the tools of bioinformatics.Among the obtained 10 512 ESTs which could be further analyzed,9,866 were from umbilical cord blood where 4 697(67.6%)were known genes,2 603(26.4%)were known ESTs and 1415(14.3%)represented novel ESTs.Within the identified genes,8.2% was involved in hematopoiesis,22.7% was associated with cell metabolism,structure and mobility,13.0% was linked to cell division and defence,26.2% was related to RNA protein synthesis and 10.6% was related with cell signal transduction.In parallel,we developed an efficient working system combining sequencing,bioinformatics,etc.and obtained 23 full-length cDNAs from both known and novel ESTs identified in this work.     

激光治疗与巨细胞包涵体病毒的临床观察

采用脱落细胞中找巨细胞包涵体病毒的测定法,检测222例慢性子宫颈炎病人的宫颈管分泌物。结果在激光术前76例阳性,其阳性率为31.5％,激光术后3～6个月中随访,72人转阴性,转阴率为94.7％。提示CO_2烧灼激光及YAG热凝固激光均能对巨细胞包涵体病毒有直接的杀灭作用。     

应用大规模测序技术和生物信息学研究造血干/祖细胞的基因表达及新基因的识别和克隆

从新生儿脐血和成人骨髓中分选出造血干／祖细胞（ＨＳＣ／ＨＰＣ）,构建成ｃＤＮＡ文库,对其进行大规模表达序列标签（ＥＳＴ）测序,通过生物信息学等手段分析基因表达谱,并进行新基因的全长ｃＤＮＡ克隆。在所测的１０５１２条可分析ＥＳＴ序列中,有９８６６条来自脐血ＣＤ３４＋细胞,其中４６９７条（４７６％）为已知基因,２６０３条（２６４％）为已知ＥＳＴ,１４１５条（１４３％）代表未知ＥＳＴ。在已知基因中,８２％基因与造血相关,２２７％涉及细胞代谢、结构和迁移,１３０％与细胞分裂和防御相关,２６２％与ＲＮＡ、蛋白质的合成相关,１０６％和细胞信号传递有关。对一些已知和未知的ＥＳＴ,综合测序、生物信息学等方法,进行全长克隆,已获得２３个新基因的全长ｃＤＮＡ。