Cisplatin-induced genes as potential markers for thyroid cancer

Despite the uncontested role of p53 in cycle arrest/cell death after cisplatin treatment, to date the question whether wild-type p53 confers a resistant or sensitive status on the cell is still a matter of debate. Isogenic and isophenotypic human thyroid papillary carcinoma cell line variants for p53 differently expressed cycle genes after cisplatin treatment. Seven genes (CDC6-related protein, CCNC, GAS1, TFDP2, MAPK10/JNK3, WEE1, RPA1) selected after expression on an Atlas human cell cycle array were analyzed by quantitative real-time PCR. While cisplatin treatment increased their expression in p53 wild-type cells it decreased it in cells with inactivated p53 and had no or less effect on cells with mutated p53. These results show that in a well-defined system, different alterations of p53 can lead to a different regulation of genes and hence to either resistance or sensitivity to cisplatin. Moreover for the first time, MAPK10/JNK3 was identified in human thyroid cells and tissue. Four of the genes (CDC6-related protein, CCNC, GAS1 and TFDP2) were decreased in human papillary carcinoma tissues. Relevance of these genes (especially a decrease in GAS1 in thyroid papillary carcinoma) in various malignant pathologies has already been shown. These genes may be explored as new markers in advanced thyroid cancer such as metastatic and anaplastic forms displaying p53 alterations.Received 26 July 2004; received after revision 14 September 2004; accepted 26 October 2004     

基因枪法导入cyclAt和GUS基因获得转基因水稻

以cyclAt基因为目的基因,HPT和GUS为选择和报告基因,应用PDS-1 000/He型基因枪协同转导,通过轰击来源于水稻成熟种子诱导产生的胚性愈伤组织,获得了转基因水稻植株.结果表明:通过X-Gluc染色分析,GUS基因在愈伤组织和再生植株叶片中的表达效率高.以转基因植株的基因组DNA为模板,用cyclAt基因的特异性引物对其进行PCR扩增,只检测到1 000 bp左右的片段,比其1 604 bp的完整序列小,因此推测cyc1At基因在转导入水稻后被剪接.此外,在协同转导过程中,cyc1At和GUS可能与受体基因组DNA随机整合,因此视GUS基因为报告基因有一定局限性;但两者同时被整合的频率远高于单一基因的整合.     

逆转录多聚合酶链反应检测排斥反应期移植肠基因的转录

排斥反应仍色是阻碍小肠移植成功的主要障碍,其机理、诊断及治疗均有待进一步研究。目前建立逆转录多聚合酶链反应方法,观察排斥反应期移植肠ＩＬ－２、ＩＦＮ－γ、穿孔素、粒细胞酶Ｂ的ｍＲＮＡ转录变化,取大鼠移植肠杆本提取总ＲＮＡ,并进行ＲＴ－ＰＣＲ检测ＩＬ－２、ＩＦＮ－γ穿孔素、粒细胞酶Ｂ的ｍＲＮＡ转录,ＰＣＲ产物以内参基因β－ａｃｔｉｎ为内对照,应用ＳＸ－１００生物影像分析系统进行半定量分析。结果显示Ｒ     

造血重建小鼠骨髓中成纤维细胞的起源

选择Y染色体特异的性别决定基因(Sry)作为新的细胞遗传标志,采用PCR技术对小鼠骨髓细胞重建造血的性能和造血重建小鼠骨髓中成纤维细胞的起源进行研究.将正常雄鼠骨髓细胞输注给经致死剂量射线照射的受体雌性小鼠,PCR技术检测结果表明,在活存小鼠的骨髓、脾脏、胸腺和淋巴结中均具有供体起源的细胞.而正常雄鼠或5-氟尿嘧啶处理的雄鼠骨髓细胞输注给受体雌性小鼠后,造血重建小鼠骨髓中的成纤维细胞则为受体起源.由此可见,小鼠骨髓中的成纤维细胞与造血细胞具有不同的起源.     

Efficient selection of homozygous lines of hybrid rice restorer with the transgeneXa21 using test cross and PCR

The homozygous restorer lines with a single copy of the transgene Xa21 have been obtained from the progenies of transgenic Minghui63 and Yanhui559 plants through PCR marker-assisted selection and test cross. These homozygous transgenic restorer lines can be used to breed hybrid rice with high resistance to bacterial leaf blight.     

江苏地区汉族人群六个短串联重复基因座的群体遗传学研究

采用复合扩增、聚丙烯酰胺凝胶电泳分型和银染显示的方法,对江苏地区汉族人群中111个无关个体的6个短串联联重复（STR）基因座（CSF1,TPOX,TH01,D16S539,D7S820和D13S317）进行了群体遗传学研究。经X^1检验。6个基因座的基因型频率符合Hardy-Weinberg平衡定律（P＞0．05）。它们的联合个体识别率（DP）值＞0．9999;联合非父排除率（PE）值＞0．9882。这6个基因座在江苏地区汉族人群中显示了高度的多态性和良好的鉴别能力,在法医学个人识别及亲子鉴定中有很高的应用价值。     

普通野生稻中NBS同源序列的克隆和分析

已知的植物抗病基因大多数具有核苷结合位点（NBS）和富含亮氨酸重复（LRR）。根据NBS的保守区设计引物,应用聚合酶链式反应（PCR）,从普通野生稻基因组中克隆了4个不同的NBS同源序列（OSNBA1－OSNBA4）。同源性比较显示它们均与非TIR类的NBS序列相似,其中,OSNBA1与双子叶植物拟南芥的RPS2基因更接近;OSNBA2－OSNBA4与单子叶植物水稻的XA1、RPR1基因更相似。     

人胎盘G-蛋白Rab3a cDNA的克隆

为了获取全长的小分子G-蛋白Rab3a,以用于研究Rab3a与其他蛋白相互作用关系,本实验以人胎盘总cDNA为模板,PCR扩增到人Rab3a cDNA全编码区。产物回收后克隆于质粒pYESTrp2的BamHI/XhoI位点,测序结果表明,本实验获得的Rab3a cDNA包含了起始和终止密码子。与PCR引物设计的参照Rab3a比较有5个核苷酸变异,与翻译的氨基酸序列完全一致。由此表明本实验获得的Rab3a cDNA可用于进一步研究。     

Molecular evolution of the exon 2 of CHS genes and the possibility of its application to plant phylogenetic analysis

The exon 2 of chalcone synthase (CHS) gene is relatively conserved during evolution. In this study, three exon 2 fragments from two species in gymnosperm (Cycas panzhihuaensis, Ginkgo biloba) and seven from four species in angiosperm (Magnolia denudata, Salix babylonica, Nymphaea tetragona, Camellia japonica) have been amplified by PCR from genomic DNA and sequenced. Together with other 73 sequences ofCHS collected from EMBL database and literature, these sequences, which embrace 19 families of gymnosperm and angiosperm, have been analyzed for their phylogenetic relations by parsimony method. The result indicated that sequences from the same systematic family usually grouped together except those from Theaceae, Magnoliaceae and Nymphaeaceae. The relative rate test revealed the rate heterogeneity of CHS genes among the families. For the nucleotide substitution the sequences from Asteraceae and Solanaceae evolve faster than those from the other families analyzed while the sequences from Poaceae, Asteraceae and Solanaceae evolve faster for the nonsynonymous substitution. These results suggest that the duplication and extinction events of CHS genes are different among systematic families, therefore it seems impractical to look for orthologous sequences from CHS genes to study plant phylogeny at the family level andlor above. However, it is possible to do so below the family level.     

AP1000汽水分离再热器(MSR)管子管板胀接技术研究及应用

本文根据国产首台AP1000 MSR管子管板胀接的制造工艺过程,从胀接设备及工具选型、工艺试验、工艺参数选定、产品胀接应用等方面进行了详细描述,为类似核电站产品的制造提供了经验借鉴。