RAPD分析─鉴定柑桔体细胞杂种的快速方法

本文利用改进的DNA提取方法,从Volkamer柠檬(Citrus volkameriana Ten. and Pasq.)和酸橙(C. aurantium L.)及其原生质体杂种植株的叶片中抽提总DNA,进行RAPD(Random Amplified Polymorphic DNA)分析。结果表明: 在随机选取的15种引物中,有10种可单独或与其它引物一道鉴定这一组合的体细胞杂种。与形态学性状观察、同工酶及ONA杂交分析等方法比较,RAPD分析是一种可在试管苗期即可直接、准确、快速鉴定柑桔体细胞杂种的方法。     

73例中国人血友病甲基因突变的分析

我们用Southern blotting、PCR、变性梯度凝胶电泳(DGGE)和DNA测序等方法对73例血友病甲患者(经上海瑞金医院测定血浆FVⅢ:C和vWF:Ag诊断,其中无亲缘关系患者65例,按FVⅢ:C水平分为轻、中、重三型。FVⅢ:C< 2%为重型,共47例;FVⅢ:C 2%-5%为中型,共9例;FVⅢ:C5%-25%为轻型,共1 7例)进行FVⅢ基因突变检测。共检出内含子22倒位23例,均为重型,约占重型的49%,与国外报道相似。余下50例(其中无亲缘关系者45)用PCR-DGG E分析所有外显子及其侧翼内含子序列,发现异常条带则进行DNA测序。在17例患者中检出突变13种,其中无义突变5种,均为重型;错义突变6种,除1例外都是轻中型;小缺失2例,都是重型;其中,AA466Lys(AAG)-Thr(ACG)、719Tyr(TAC)-Stop(TAG)、AA826 Asp(GAC)-Glu(GAA)、312Ile(ATC)-xxC及AA1551-1552del(AGAA)为新发现的突变。有亲缘关系的患者都有相同的基因突变,而在无亲缘关系患者未发现相同突变。基因突变与临床表现基本相符。 Abstract:We use Southern blotting,PCR,denaturing gradient gel electrophoresis(DGGE)and DNA sequencing to detect gene mutations of haemophilia A in Chinese population.73 cases(47 severe)(FVIII:C<2%),9 moderate(FVIII:C 2%~5%),17 mild(FVIII:C 5%~25%)of haemophilia A were first screened with Southern blotting,23 were found to be the intron 22 inversion type,all being severe cases.The remaining 50 cases without intron 22 in version were examined with PCR-DGGE.Genomic DNA were amplified using GC-clamped primers covering all the exons and all flanking intron regions.Abnormal bands were sequenced.13 different mutations were identified,including 5 nonsense mutations,6 missense mutations and 2 small deletions.5 mutations,AA466Lys(AAG)-Thr(ACG),AA719Tyr(TAC)-Stop(TAG),AA826Asp(GAC)-Glu(GAA),AA312Ile(ATC)-xxC and AA1551-1552del(AGAA)have not been reported before.Generally the genetic defects correspond to the clinical conditions.     

大熊猫与小熊猫生态习性的比较：食物、体型大小及系统发育的影响

大熊猫和小熊猫是食肉目中尤为引人注目的 两个物种,均特化为以竹类为生,在四川西部的横断山地同域共存.近年来化石和分子生物学等方面的证据表明两者亲缘关系较远,其在诸多形态特征上的相似乃是长期趋同进化的结果.竹子是一类低营养价值的食物,有丰富的储量及相对稳定的营养组成,保障了两种熊猫的长期续存.由于仍保留着食肉动物消化系统的典型特征,大熊猫、小熊猫对食物的消化利用率极低,两者在一些生态习性(如独栖、无冬眠、巢域普遍重叠及具有活动与休息多次交替的昼夜活动模式等)的相似可能反映了这种特殊食物的影响.此外,大熊猫和小熊猫在食性、昼夜活动节律及微生境利用上的差异与其体型大小及系统发育有关,有利于两者在川西横断山地的同域共存.     

Adenovirus-mediated expression of pig α(1,3) galactosyltransferase reconstructs Gal α(1,3) Gal epitope on the surface of human tumor cells

INTRODUCTIONGal a(1, 3) Gal (gal epitope) is a carbohydrate epitope, which is produced in large amounton the cells of pigs, mice and New World monkey(monkey of South America) by the glycosylationenzyme G alal 1 ) 4G IcNAc3- a- D- galactosyltransferase[or(1, 3)GT; EC2.4.1.511111. This enzyme is active in the Golgi appaxatus of cells and transfers galactose from the sugandonor uridine diphoSphate galactose (UDP-galactose) to the acceptor Nacetyllactosamine residue (Galaal-4GlcNAc-R…     

双歧杆菌对新生大鼠坏死性小肠结肠炎肠损伤及肠道菌群影响

目的建立新生SD大鼠坏死性小肠结肠炎（NEC）模型,探讨添加双歧杆菌对新生大鼠NEC模型肠损伤的保护作用及肠道菌群的影响,为应用双歧杆菌防治NEC提供依据：方法SD新生大鼠出生48h开始给予鼠配方奶人工喂养,100％氮气缺氧90s,4℃冷刺激10min,每天2次,连续3d,建立新生SD大鼠NEC模型。按析因设计,32只新生SD大鼠随机分成4组,每组动物各8只。A组为NEC模型组并在出生48h起每日给予长双歧杆菌灌胃（1×10^8CFU／d）;B组为NEC模型组,C组为对照组,都未添加双歧杆菌;D组为对照组并给予长双歧杆菌灌胃（1×10^8CFU／d）。在最后一次缺氧、冷刺激后24h空腹断头处死大鼠,留取回盲部近端肠管组织进行肠组织损伤评分,组织学评分≥2确定为NEC;实验前后留取各组新生鼠粪便,按照张秀荣方法进行肠道菌群检测。应用Kruskal-Wallis H检验等方法进行统计学分析,α=0．05为显著性检验标准。结果开始造模后,A、B组新生SD大鼠相继出现腹泻、腹胀、萎靡、活动减少,A组程度较轻。A、B、C和D组肠组织损伤评分（^-x±s）分别为1．88±0.84、3．13±0．84、0．63±0．52、0．50±0.54,各组间肠组织损伤评分差异有显著性,与B组相比,A组肠组织损伤评分明显降低,但仍高于C、D两组,差异均有显著性。各组实验前肠道细菌总数,杆菌、球菌数,G^＋杆菌、G^＋球菌数差异均无显著性,肠道菌群中杆球菌比值在正常范围中。实验结束时,各组间新生鼠肠道细菌总数,杆菌、球菌数,杆球菌比值,G^＋杆菌、G^＋球菌数及其占肠道总菌群数的比率差异均有显著性;除B组杆菌数外,与实验前相比,各组新生大鼠实验结束后肠道细菌总数、球菌数、杆菌数、G^＋杆菌数、G^＋球菌数均有显著增加,差异均有显著性;A、B组肠道杆球菌比值和G^＋杆菌数占肠道总菌群数比     

不同化学杂交剂(CHA)对小麦花药同工酶影响的研究

在小麦叶枕距±2cm时,喷施4种不同化学杂交剂（CHA）后,分别取小孢子处于不同发育阶段的花药,进行过氧化酶（POD）、淀粉酶（Amy）和酯酶（Est）等同工酶的分析。研究表明：在小孢子母细胞减数分裂期,处理2、3和4三种CHA对POD的A1、A2;B2、B4和B5,C1、C2和C3同工酶带的活性均有明显的抑制作用;处理5除去对A1和A2表现抑制外,对其他酶带的活性均有增强作用。在单枚早期,处理2和3的A、B和C区POD同工酶活性均明显低于对照;处理4和5上述各区POD同工酶活性却明显高于对照。在上述两个发育时期,处理2对Amy1区酶活性有增强作用,而处理3、4和5对该区酶活性却表现了专一性的抑制。各处理对Est同工酶A区和B区的酶活性主要表现为抑制。实验结果表明,不同的CHA均通过干扰花药的物质和能量代谢而导致雄性生理性不育。     

Comparative systems biology between human and animal models based on next-generation sequencing methods

Animal models provide myriad benefits to both experimental and clinical research. Unfortunately, in many situations, they fall short of expected results or provide contradictory results. In part, this can be the result of traditional molecular biological approaches that are relatively inefficient in elucidating underlying molecular mechanism. To improve the efficacy of animal models, a technological breakthrough is required. The growing availability and application of the high-throughput methods make systematic comparisons between human and animal models easier to perform. In the present study, we introduce the concept of the comparative systems biology, which we define as "comparisons of biological systems in different states or species used to achieve an integrated understanding of life forms with all their characteristic complexity of interactions at multiple levels". Furthermore, we discuss the applications of RNA-seq and ChIP-seq technologies to comparative systems biology between human and animal models and assess the potential applications for this approach in the future studies.     

巴西人参化学成分和药理活性研究进展

巴西人参原产于南美洲,当地人把它当做具有壮阳、增强体质等作用的草药。巴西人参已在我国广西、浙江等地引种栽培成功,现代研究发现,其主要含有三萜和三萜皂苷类、甾体类等多种化学成分,具有抗肿瘤、抗炎、保护胃黏膜等药理活性。本文通过对近年来巴西人参的研究情况进行综述,阐明已有的化学物质基础和药理作用,为其进一步的深入研究和开发提供理论依据。     

抗癌药对黑胸大蠊淋巴细胞和生殖细胞核结构的损伤

噻替派浓度为0.1%、0.3%、0.5%时,黑胸大蠊精母细胞染色体断裂和裂隙率分别为6.3%、 10.5%和14.2%,显著地高于对卵母细胞的影响;和雄虫外周血淋巴细胞微核率呈平行关系,随微核率增多而增加。5-氟尿嘧啶浓度为0.1%、0.3%和0.5%时,卵母细胞染色体断裂和裂隙率分别为3.5%、9.8%和16.2%,和雌虫外周血淋巴细胞微核率呈平行关系,随微核率增多而增加,而对雄虫生殖细胞影响不显著。 Abstract:0.1%,0.3%,0.5% Thio-TEPA induced 6.3%,10.5% and 14.2% chromosome break or gap in spermatocyte of cockroach respectively.This was markedly higher than those in oocyte.In doses from 0.1 to 0.5 Tho-TEPA the frequency of micronucleus increased parallely with nuclear damage.0.1%,0.3%,0.5% 5-fluorouracil induced 3.5%,9.8%,16.2% chromosome break or gap in oocytes respectively.This was paralled with the frequency of micronucleus in lymphocytes of the female.5-fluorouracil showed not marked effect on spermatocyte.