Cystic fibrosis locus defined by a genetically linked polymorphic DNA marker

A polymorphic DNA marker has been found genetically linked, in a set of 39 human families, to an autosomal recessive gene that causes cystic fibrosis (CF), a disease affecting one in 2000 Caucasian children. The DNA marker (called D0CRI-917) is also linked to the PON locus, which by independent evidence is linked to the CF locus. The best estimates of the genetic distances are 5 centimorgans between the DNA marker and PON and 15 centimorgans between the DNA marker and the CF locus, meaning that the location of the disease gene has been narrowed to about 1 percent of the human genome (about 30 million base pairs). Although the data are consistent with the interpretation that a single locus causes cystic fibrosis, the possibility of genetic heterogeneity remains. The discovery of a linked DNA polymorphism is the first step in molecular analysis of the CF gene and its causative role in the disease.     

Genetic mechanisms of tumor suppression by the human p53 gene

Mutations of the gene encoding p53, a 53-kilodalton cellular protein, are found frequently in human tumor cells, suggesting a crucial role for this gene in human oncogenesis. To model the stepwise mutation or loss of both p53 alleles during tumorigenesis, a human osteosarcoma cell line, Saos-2, was used that completely lacked endogenous p53. Single copies of exogenous p53 genes were then introduced by infecting cells with recombinant retroviruses containing either point-mutated or wild-type versions of the p53 cDNA sequence. Expression of wild-type p53 suppressed the neoplastic phenotype of Saos-2 cells, whereas expression of mutated p53 conferred a limited growth advantage to cells in the absence of wild-type p53. Wild-type p53 was phenotypically dominant to mutated p53 in a two-allele configuration. These results suggest that, as with the retinoblastoma gene, mutation of both alleles of the p53 gene is essential for its role in oncogenesis.     

Alterations in 5-HT1B receptor function by p11 in depression-like states

The pathophysiology of depression remains enigmatic, although abnormalities in serotonin signaling have been implicated. We have found that the serotonin 1B receptor [5-hydroxytryptamine (5-HT1B) receptor] interacts with p11. p11 increases localization of 5-HT1B receptors at the cell surface. p11 is increased in rodent brains by antidepressants or electroconvulsive therapy, but decreased in an animal model of depression and in brain tissue from depressed patients. Overexpression of p11 increases 5-HT1B receptor function in cells and recapitulates certain behaviors seen after antidepressant treatment in mice. p11 knockout mice exhibit a depression-like phenotype and have reduced responsiveness to 5-HT1B receptor agonists and reduced behavioral reactions to an antidepressant.     

Mapping of Microsatellite SW943 to Porcine Chromosome 12p11-(2/3p13) Using Primed in situ Synthesis and Somatic Cell Hybrid Panel

The porcine microsatellite SW943 was regionally localized on 12p11-(2/3p13) by the two methods: the Primed in situ (PRINS) labelling on the pachytene bivalents of pigs using the Dig-11-dUTP as the report molecule and pig × rodent Somatic Cell Hybrid PaneI(SCHP) which contains 27 cell lines through PCR amplification. Advantages and disadvantages of the two methods for physical mapping of microsatellites were also discussed.     

11-13岁少年儿童体质健康促进效果的研究

目的：根据少年儿童身心发育特点,设计符合学校开展的健身干预方案,通过实施,探索运动干预方案对学生体质的影响。方法：选取西安市某小学5年级8个班男、女生388人为实验对象,实施运动干预。实验前后对受试者相关指标进行测试,测试指标选取身体形态、机能和素质三类指标14项测定并进行分析,显著性取P<0.05。结果：10周的运动干预,使受试者体脂含量、心肺功能、力量、柔韧、灵敏性、平衡稳定性及反应速度等方面都出现了显著变化（P<0.05）。通过干预提高了受试少年儿童的身体机能和身体素质水平。结论：运动干预方式能够有效提高少年儿童参与体育锻炼的兴趣和学生整体体质健康水平,促进身体素质的全面发展。干预方案可以作为学校组织开展体育课及阳光体育活动、促进学生体质增长的实践方式。     

ER cargo properties specify a requirement for COPII coat rigidity mediated by Sec13p

Eukaryotic secretory proteins exit the endoplasmic reticulum (ER) via transport vesicles generated by the essential coat protein complex II (COPII) proteins. The outer coat complex, Sec13-Sec31, forms a scaffold that is thought to enforce curvature. By exploiting yeast bypass-of-sec-thirteen (bst) mutants, where Sec13p is dispensable, we probed the relationship between a compromised COPII coat and the cellular context in which it could still function. Genetic and biochemical analyses suggested that Sec13p was required to generate vesicles from membranes that contained asymmetrically distributed cargoes that were likely to confer opposing curvature. Thus, Sec13p may rigidify the COPII cage and increase its membrane-bending capacity; this function could be bypassed when a bst mutation renders the membrane more deformable.     

Effect of p53 status on tumor response to antiangiogenic therapy

The p53 tumor suppressor gene is inactivated in the majority of human cancers. Tumor cells deficient in p53 display a diminished rate of apoptosis under hypoxic conditions, a circumstance that might reduce their reliance on vascular supply, and hence their responsiveness to antiangiogenic therapy. Here, we report that mice bearing tumors derived from p53(-/-) HCT116 human colorectal cancer cells were less responsive to antiangiogenic combination therapy than mice bearing isogenic p53(+/+) tumors. Thus, although antiangiogenic therapy targets genetically stable endothelial cells in the tumor vasculature, genetic alterations that decrease the vascular dependence of tumor cells can influence the therapeutic response of tumors to this therapy.     

用引物原位合成和体细胞杂种克隆板将猪微卫星SW943定位于12p11～(2/3p13)

 以Dig 11 dUTP为报告分子 ,运用猪微卫星SW 94 3引物在猪减数分裂粗线期二价体 (下称二价体 )上进行引物原位DNA合成 (primedinsitulabeling ,PRINS) ,用鼠抗地高辛、兔抗鼠、羊抗兔抗体检测引物原位合成结果对SW 94 3进行定位研究 ,同时运用包含猪 啮齿类 2 7个细胞系的杂种细胞克隆板 ,通过PCR扩增进行SW 94 3的定位 ,结果将猪微卫星SW 94 3定位于 12 p11～ (2 /3p13)。     

miR-221-3p靶向BCL2L11调控小尾寒羊卵泡颗粒细胞凋亡

【目的】BCL2L11在哺乳动物中能够促进多种细胞凋亡,同时参与繁殖性状相关组织器官的发育及疾病治疗,文章利用分子生物学方法探究miR-221-3p靶向调控BCL2L11对小尾寒羊卵泡颗粒细胞凋亡的影响,为进一步研究BCL2L11在卵泡颗粒细胞凋亡和卵泡闭锁过程中的调控作用提供依据。【方法】在前期课题组卵巢组织全转录组测序分析的基础上,获得了候选基因BCL2L11及其调控元件miR-221-3p,利用半定量和组织荧光定量（RT-qPCR）分析BCL2L11在小尾寒羊不同组织中的表达情况;通过RT-qPCR定量试验在小尾寒羊卵泡期和黄体期卵巢组织中鉴定了BCL2L11及miRNA-221-3p的表达情况;构建BCL2L11 3’UTR野生型和突变型载体,在HEK293T细胞中共转染miR-221-3p mimic和BCL2L11野生型和突变型及阴性对照,采用双荧光素酶报告基因检测系统确定miR-221-3p与BCL2L11靶向性关系;在绵羊卵巢原代颗粒细胞中转染miR-221-3p mimic及阴性对照实现miR-221-3p过表达,使用RT-qPCR技术在mRNA水平上检测miR-221-3p对BCL2L11以及卵巢颗粒细胞凋亡标志基因XIAP和Fas表达水平的影响;同时利用EdU试验分析miR-221-3p过表达和阴性对照组中颗粒细胞的增殖变化。【结果】半定量和组织RT-qPCR分析均表明BCL2L11在卵巢组织中表达量高于其他组织;RT-qPCR定量结果显示miR-221-3p和 BCL2L11在小尾寒羊卵泡期和黄体期卵巢组织中差异表达,miR-221-3p在卵泡期卵巢中的表达量高于黄体期,而BCL2L11在卵泡期卵巢中的表达量低于黄体期,表现出负调控的现象;双荧光素酶报告基因验证分析显示,过表达miR-221-3p mimic显著抑制了BCL2L11 3’UTR荧光素酶的活性（P<0.05）,阴性对照组则没有显著影响;过表达miR-221-3p,靶基因BCL2L11 mRNA表达水平显著降低,同时,卵泡颗粒细胞凋亡标志基因XIAP和Fas的表达量也显著降低（P<0.05）;EdU试验分析显示,过表达miR-221-3p的颗粒细胞增殖率为18.9%,极显著高于阴性对照组的10.43%（P<0.01）。【结论】BCL2L11和miR-221-3p是调控绵羊卵巢发育的重要基因及调控元件,BCL2L11是miR-221-3p的靶基因之一,miR-221-3p过表达可抑制颗粒细胞凋亡,该作用结果可能通过抑制靶基因BCL2L11的表达进而影响了绵羊卵巢颗粒细胞的凋亡。     

体外筛选对鸡具有免疫刺激活性的CpG寡脱氧核苷酸

分别用有丝分裂原和CpG寡脱氧核苷酸(CpG ODN)对SPF鸡全血，经低速离心分离的外周血单个核细胞(PBMC)和脾脏细胞，用淋巴细胞分离液分离的PBMC和脾脏细胞进行刺激，用^3H-TdR掺入法测定并比较上述鸡免疫细胞在不同培养温度和细胞浓度下的转化效果，从而建立了适用于大批量筛选对鸡具有免疫刺激活性CpG ODN的淋巴细胞转化方法。进一步用该方法筛选自行设计合成的38条ODN.结果表明，用低速离心法分离的鸡PBMC，在细胞含量为10^7mL^-1、37℃、5％CO2条件下，经CpG ODN刺激64～66h后，可以得到稳定的高效转化；不含CpG模体的ODN不具有免疫刺激活性；CpG ODN 10、11和14具有最佳的免疫刺激活性，刺激指数大于10。     

模拟葡萄汁中添加可同化氮对酵母菌酒精发酵的影响

【目的】研究添加可同化氮对模拟葡萄汁酒精发酵的影响,确定利于酵母菌酒精发酵的可同化氮水平,为葡萄酒酿造过程中可同化氮的添加提供依据。【方法】设计3种氮素添加方式,分析不同质量浓度可同化氮和添加处理下模拟葡萄汁的酒精发酵过程及产物的差异,对各处理酒精发酵过程中的酵母菌数量和还原糖含量及酒精发酵结束后的游离氨基酸、铵态氮、有机酸、香气成分含量进行测定和分析。【结果】随着可同化氮质量浓度的升高,酵母菌数量增加,酒精发酵速率增大,二次添加可同化氮使酵母菌数量增加较快,糖消耗速率增大;可同化氮质量浓度为240mg/L时,有机酸的总生成量最高,二次添加可同化氮降低了有机酸的总生成量,提高了乳酸、乙酸和琥珀酸的生成量,但柠檬酸的生成量有所降低;可同化氮质量浓度的升高有利于生成较多的香气成分,二次添加处理可以提高高级醇和酯类的生成量,但对酸类生成量无明显影响。【结论】可同化氮是酵母菌进行正常酒精发酵必需的大量营养元素,可同化氮质量浓度低于90mg/L时,添加有机氮(氨基酸)和无机氮(铵态氮)均能促进酵母菌完成酒精发酵,可同化氮质量浓度的升高有利于生成较多的香气成分。     

Light Flashes Observed by Astronauts on Apollo 11 through Apollo 17

The crew members on the last seven Apollo flights observed light flashes that are tentatively attributed to cosmic ray nuclei (atomic number >/= 6) penetrating the head and eyes of the observers. Analyses of the event rates for all missions has revealed an anomalously low rate for transearth coast observations with respect to translunar coast observations.     

扎龙湿地11种鸟类所带细菌致病性的试验

从扎龙湿地１１种野生和笼养鸟类的口咽部分泌物,泄殖腔内容物及新鲜粪便中,曾分离出２５种不同的细菌。为了探讨这些自然携带菌的致病性。根据以往文献记载对所分离到的２５种细菌中可能具有致病性或致病性不明确的１４细菌进行了小白鼠致病性试验。     

电弧喷涂3Cr13强化秸秆压块机模块试验研究

针对秸秆压块机模块磨损失效问题,分析了模块失效机理,提出了采用电弧喷涂3Cr13丝材对秆压块机模块表面进行强化和修复新工艺。采用正交试验和极差分析法研究了喷涂电流、喷涂电压、喷涂距离、喷涂气压对涂层耐磨性的影响规律,优化了压块机模块电弧喷涂3Cr13工艺参数,测试了3Cr13涂层的摩擦系数。结果表明:涂层摩擦系数为0.36,远低于铸钢摩擦系数0.8,涂层具有优异的耐磨性。     

复合酶法制备金华火腿水解液的研究

以水解度和感官评定（滋味和气味）为指标,比较了中性蛋白酶、碱性蛋白酶、酸性蛋白酶、风味蛋白酶单一酶和复合酶对金华火腿下脚料的水解效果,通过正交实验设计,结果表明,采用中性蛋白酶和风味蛋白酶按1∶1组合,加酶量3 000 U·g-1,固液比1∶3,pH值7.5,温度50℃的条件下水解3 h,可获得适度水解、风味较佳、无苦味的水解液。     

基于SSR荧光标记的MCID快速鉴定13个湖北茶树品种

[目的]通过人工绘制品种鉴别图(Manual cultivar identification diagram,MCID)快速区分鉴别湖北茶树品种,为湖北茶树种质资源评价、品种保护及苗木纯度早期鉴定提供技术支持.[方法]以湖北省13个优良茶树品种为材料,利用30对均匀分布于遗传连锁图谱上的SSR荧光引物进行PCR扩增,并用荧光标记毛细管电泳检测扩增产物以筛选出核心引物,利用MCID法构建CID图谱.[结果]30对SSR引物共扩增出145个等位基因,各引物等位基因数为3~9个,平均每对引物为4.83个;共检测到194个基因型,各引物检测出的基因型为3~12个,平均每对引物6.47个;多态性信息量(PIC)为0.29~0.85,平均为0.58. 13个茶树品种的Nei's遗传距离为0.19~0.44.基于Nei's遗传距离,采用Neighbor-Joining(NJ)法可将13个品种分为三大类(Nei's遗传距离为0.16),结果表明地理来源相同的品种可能因为具有相似的遗传背景而聚在一起,也可能因为亲本来源不同而存在明显的遗传差异.利用筛选出的3对核心引物(TM547、TM552和TM107)可快速鉴别所有参试品种,通过MCID法构建的CID图谱可直观看出各参试茶树品种鉴定所需要的引物及其对应的基因型.[结论]基于SSR荧光标记的茶树品种MCID鉴定方法具有快速、准确、高效的特点,可用于湖北茶树良种知识产权保护、苗期鉴定及品种区分.     

穗霉属26-13-4菌株产植酸酶的酶学性质研究

从黑龙江大豆根部土壤中分离一株穗霉属菌株,对其诱变菌株26-13-4所产植酸酶进行研究。该植酸酶对植酸钠的适宜水解温度为45℃、适宜pH为5.0,在4、25和45℃下热稳定性好,在pH4.0～7.0时植酸酶比较稳定;低浓度的Fe2+、Mn2+、Ca2+对酶活性有促进作用,而Cu2+、Mg2+、Zn2+、Al3+以及高浓度Mn2+、Ca2+、Fe3+对酶活性有抑制作用;K+、低浓度的Fe3+对酶活没有任何影响。结果表明,该植酸酶以植酸钠为底物时的米氏常数Km为4.4×10-5mol·L-1。     

用13C-核磁共振方法研究有机肥料对胡敏酸结构特征的影响

通过在棕壤上进行的田间试验和模拟试验,研究了施用有机肥胡敏酸（ＨＡ）＾１３Ｃ－核磁共振（ＮＭＲ）波谱的影响。结果表明,施用有机冯有使ＨＡ的碳骨架发生规律性的变化。一般表现为芳香碳（Ａｒ－Ｃ）和羰基碳（〉Ｃ＝Ｏ）含量减少;而烷基碳（Ｒ－Ｃ）和烷氧碳（ＲＣ－Ｃ）的含量增加。从恧地致ＨＡ的脂族化。研究表明,文中建立的土壤ＨＡ溶液＾１３Ｃ－ＮＭＲ方法,用于相对比较土壤有机增减肥后ＨＡ结构特征的变化规律是可