Radiation sensitivity of lymphocytes from healthy individuals and cancer patients as measured by the comet assay

Lymphocytes of healthy volunteers (n=24) and of tumour patients (n=30, 18 of whom had experienced severe side-effects) were irradiated with x-rays in vitro. DNA damage was analysed after 0.25–2 Gy and DNA repair after 2 Gy, and quantification of both endpoints was done by the comet assay. The individual differences in radiation-induced DNA damage as well as in the repair kinetics were observed to be striking for both healthy donors and tumour patients. After a repair time of 3 h, following 2 Gy x-irradiation, some of the healthy volunteers showed no residual DNA damage at all in their lymphocytes, whereas others revealed about 30%. There was no indication that our results were affected by either age, gender or smoking habits. Slow repair kinetics and high amounts of residual damage were characteristic for many but not all tumour patients who had experienced severe side-effects in their normal tissues during or after radiotherapy (n=18). Our conclusion is that those individuals showing poor DNA repair characteristics in the lymphocytes following in vitro irradiation, have a high probability of being radiosensitive. The opposite conclusion is not necessarily true: if repair is effective, this does not mean that the individual is radioresistant, because factors other than impaired repair may cause radiosensitivity. Received: 3 May 2000 / Accepted: 1 December 2000     

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.     

用长PCR方法检测含有较大缺失或插入的DNA大片段

选择位于19q13.3上的人类肌张力蛋白激酶基因(myotonin protein kina se gene,MT-PK)为靶基因(基因全长为14kb),以G+C含量较高且含有1kb缺失或插入,由基因第8内含子中的Alu±1kb的5'端至第15外显子3'非编码区中的CTG重复序列3'端,即两者间的距离为5.3kb的DNA片段为待扩增靶序列,通过优化DNA聚合酶的组合和反应缓冲体系,点考查了含有Alu-1k b和Alu+1kb缺失或插入的MT-PK等位基因片段共扩增的长PCR方法。本方法可有效地同步扩增6.5kb和5.5kb两个等位基因片段,对6.5kb和5.5kb纯合等 位片段则达到了更有效的扩增。     

火烧对内蒙古草原中坚韧胶衣固氮活性的影响

 坚韧胶衣(Collema tenax)是干旱和半干旱草原中常见的一种固氮地衣, 是草原生态系统中生物土壤结皮(Biological soil crust)的 主要组成部分, 对生态系统氮循环具有重要的影响。火烧作为一种干扰因子, 是草原生态系统结构和功能维持的重要因素之一。该文采用乙炔 还原法(Acetylene reduction assay), 研究了火烧对内蒙古草原生态系统中坚韧胶衣固氮活性的短期影响。结果表明, 在个体尺度上, 与对照 相比, 火烧区中地衣体烧损的坚韧胶衣固氮活性降低了42.3%, 而无烧损的个体固氮活性则升高了28.4%。这表明火烧对坚韧胶衣的固氮功能在 个体尺度上具有双重影响: 1)通过烧损地衣体、恶化地表温度和水分条件, 而抑制个体的固氮活性; 2)通过改善光照条件, 使表土养分呈现脉 冲式增高, 而促进未烧损个体的固氮活性。在种群尺度上, 火烧与对照之间固氮活性并无显著差异, 这可能是由于火烧在个体尺度上对坚韧胶 衣的固氮活性的双重影响相互抵消所致。     

Rescue and Preliminary Application of a Recombinant Newcastle Disease Virus Expressing Green Fluorescent Protein Gene

Based on the complete genome sequence of Newcastle disease virus (NDV) ZJI strain, seven pairs of primers were designed to amplify a cDNA fragment for constructing the plasmid pNDV/ZJI, which contained the full-length cDNA of the NDV ZJI strain. The pNDV/ZJI, with three helper plasmids, pCIneoNP, pCIneoP and pCIneoL, were then cotransfected into BSR-T7/5 cells expressing T7 RNA polymerase. After inoculation of the transfected cell culture supernatant into embryonated chicken eggs from specific-pathogen-free (SPF) flock, an infectious NDV ZJI strain was successfully rescued. Green fluorescent protein (GFP) gene was amplified and inserted into the NDV full-length cDNA to generate a GFP-tagged recombinant plasmid pNDV/ZJIGFP. After cotransfection of the resultant plasmid and the three support plasmids into BSR-T7/5 cells, the recombinant NDV, NDV/ZJIGFP, was rescued. Specific green fluorescence was observed in BSR-T7/5 and chicken embryo fibroblast (CEF) cells 48h post-infection, indicating that the GFP gene was expressed at a relatively high level. NDV/ZJIGFP was inoculated into 10-day-old SPF chickens by oculonasal route. Four days post-infection, strong green fluorescence could be detected in the kidneys and tracheae, indicating that the recombinant GFP-tagged NDV could be a very useful tool for analysis of NDV dissemination and pathogenesis.     

不吸水链霉菌梧州新亚种基因组文库的构建

以不吸水链霉菌梧州新亚种为材料,提取的总基因组DNA经Sau3 AI不完全酶切。回收20-30kbDNA酶切片段,与经Hpal和BamHI酶切的粘粒载体pKC505进行连接,将连接产物用噬菌体包装蛋白包装。侵染大肠埃希菌DH5α。在舍有安普霉素的LB平板上培养,所得转化子数为12000个,构建成不吸水链霉菌梧州新亚种的基因组文库。以不吸水链霉菌染色体基因组大小为7Mb计算,概括了不吸水链霉菌梧州新亚种约99％的基因组,达到了建库要求的理论值。未扩增文库的滴度为1．2×10^8pfu／L,扩增文库的滴度为4．8×10^11pfu／L,包装效率为每微克DNA1．2×10^5个转化子。随机挑取菌落,提取重组质粒进行酶切电泳分析,均有外源片段插入。基因文库的构建为进一步深入研究梧宁霉素生物合成基因结构及功能奠定基础。     

Nonlinear Phenomena Conveying Body Size Information and Improving Attractiveness of the Courtship Calls in the Males of Odorrana tormota

Nonlinear phenomena are commonly shown in the vocalization of animals and exerts different adaptive functions.Although some studies have pointed out that nonlinear phenomena can enhance the individual identification of male Odorrana tormota,whether the nonlinear phenomena play a specific role in the sexual selection of O.tormota remain unclear.Here we presented evidence that there was a significant negative correlation(Pearson:n=30,r=0.65,P<0.001)between the nonlinear phenomena content and snout-vent length in the male O.tormota,and two-choice amplexus experiments showed that female O.tormota preferred male with smaller body size containing higher nonlinear phenomena content in its calls.Phonotaxis experiments also revealed that females preferred calls with higher nonlinear phenomena content.Additionally,compared to the calls with lower nonlinear phenomena content and higher fundamental frequency,there was shorter response time in phonotactic behaviour of female induced by the calls with higher nonlinear phenomena content and lower fundamental frequency.We argue that the nonlinear phenomena content in the calls of male O.tormota can convey its body size information and may provide important clues for female frogs in darkened surroundings to identify males’body size during mate choice,meanwhile,higher nonlinear phenomena content in males’calls may increase the attractiveness of males to females.The results of this study provide confirmation that,for O.tormota,nonlinear phenomena have specific function in mate choice.     

离体条件下用秋水仙素处理小麦-黑麦杂种细胞的研究

在液体培养基上用0.05%的秋水仙素处理小麦-黑麦杂种幼穗愈伤组织,含秋水仙素的培养基采用两种方法灭菌：（1）秋水仙素与培养基成分混合在一起用灭菌锅灭菌。处理愈伤后愈伤组织无伤害迹象,推测可能是灭菌过程中秋水仙素浓度降低所致;再生植株的育性得到有效的恢复,48.1%的穗子结实,5.2%的穗子结实率达到34.8%;（2）秋水仙素经超滤灭菌后,在常温下与灭过菌的培养基成分混和,处理愈伤后愈伤组织伤害严重,几乎不能分化苗。     

Distribution of gamasid mites on small mammals in Yunnan Province, China

An investigation of gamasid mites on the body surface of small mammals was carried out in Yunnan Province of China from 1990 to 2004. The small mammal hosts were captured from 25 counties which represent five geographical subregions, namely Middle Subregion of Hengduan Mountains, Southern Subregion of Hengduan Mountains, Eastern Plateau Subregion of Yunnan, Western Plateau Subregion of Yunnan and Southern Moun- tainous Subregion of Yunnan. The captured 10 803 small mammal hosts belong to nine families, 29 genera and 52 species in four orders （Rodentia, Insectivora, Scandentia and Lagomorpha）. A total of 68 571 gamasid mites were collected from the body surface of the captured small mammal hosts and all the gamasid mites were identified to 10 families, 33 genera and 112 species. This paper lists all the mite species, together with their taxonomic position （genera and families） and their corresponding hosts. Much more mite species were found in the Middle Subregion of Hengduan Mountains than in other geographical subregions. The total individuals of mites and small mammals in the Middle Subregion of Hengduan Mountains are also the most plentiful in the five geographical subregions. Three dominant mite species and three dominant small mammal hosts were determined as the dominant species in the investigated areas of Yunnan Province. The dominant hosts are Rattus flavipectus （which accounts for 34.85% of the total individuals）, Apodemus chevrieri （13.43%） and Rattus norvegicus （10.40%） while the dominant gamasid mite species are Laelaps nuttalli （Hirst, 1915） （27.84%）, Laelaps echidninus （Berlese, 1887） （18.38%） and Laelaps guizhouensis （Gu et Wang, 1981） （14.79%）. The results showed the high species diversity of gamasid mites in Yunnan Province and the uneven distribution feature in different subregions.     

Nuclear translocation of EGF receptor regulated by Epstein-Barr virus encoded latent membrane protein 1

Epstein-Barrvirus(EBV),oneoftheDNAon-cogenicviruses,iscloselyassociatedwiththegenesisofBurkitt抯lymphoma,undifferentiatednasopharyn-gealcarcinoma(NPC),Hodgkin抯disease,gastriccancerand,lungcancer,etc.[1].EBVencodedlatentmembraneprotein1(LMP1)isconsideredtobethemajoroncogenicproteinofEBVencodedproteins.Biologicallyspeaking,LMP1isanintegralmembraneproteincontaining386aminoacids.Thethreedo-mains(CTAR1,CTAR2,CTAR3)intheC-terminusofLMP1havebeenshowntoinitiatethesignalingproc-ess.The…