基于cytb基因分析帕米尔盘羊和新疆地方家绵羊的系统发育

[目的]研究帕米尔盘羊对新疆地方家绵羊是否有遗传贡献。[方法]测定了盘羊、家绵羊75个体的细胞色素b基因全序列,分析其遗传多样性,计算不同群体间遗传距离,进而构建系统发生树。[结果]新疆地方绵羊中检测到43个多态位点,定义为23个单倍型,单倍型多样度(Hd)为0.889±0.00102,核苷酸多样度(Pi)为0.521%。新疆地方家绵羊群体可分为4个支系(A、B、C和D),其中支系D首次在新疆多浪羊群体中被检测到。[结论]系统发育分析表明,新疆绵羊有多个母系起源或经过多次驯化,摩弗伦羊是绵羊母系祖先之一。     

绵羊Cytb基因序列多态性及系统进化研究

以8个中国地方绵羊品种和1个外来品种的20个个体为研究对象,通过对Cytb基因的全序列测定,结果表明：绵羊的单倍型多样性为97．1％。所有序列的平均碱基组成为27．1％T,28．5％C,31．4％A及13．0％G,G＋C含量为41．5％,核苷酸多样性为0．602％。在所有序列中共检测到43个变异位点,其中包括40处转换和3处颠换。Fu’S中性检验表明差异不显著（0．10〉P〉0．05）,说明绵羊群体未发生群体扩张事件。NJ、ME及UPGMA聚类结果均表明,我国绵羊可分为3个单倍型组,这提示我国绵羊有3个母系起源。     

中国山羊mtDNA D-loop遗传多样性及其起源研究

采用DNA测序技术分析了中国9个山羊品种（板角山羊、成都麻羊、贵州黑山羊、贵州白山羊、黔北麻羊、马头山羊、陕南白山羊、黄淮山羊和雷州山羊）共计128个个体的mtDNA D-loop全序列。结果表明：山羊mtDNA D-loop全序列长度为1212-1213bp,检测到102个变异位点,约占分析位点总数的8．42％,可变位点中转换占99个,颠换2个,1个转换/颠换共存;界定了92种单倍型,有78种为各品种独享单倍型,另外14种为群体内或群体间共享单倍型。9个山羊品种单倍型多样度为0．9333-1．0000,核苷酸多样度为0．7062％-1．8265％,表明中国山羊品种遗传多样性丰富。根据92种mtDNA单倍型构建了中国山羊的NJ分子系统树,聚类表明,中国山羊mtDNA D-loop序列单倍型分为支系A和支系B两大类。支系A包括75种单倍型,代表95个样本,占总数的74．22％;支系B包括17种单倍型,代表33个样本,占总数的25．78％,说明中国山羊存在支系A和支系B两大母系起源。对中国山羊mtDNA D-loop的支系A和支系B进行核苷酸不配对分布曲线分析和Fu的Fs中性检验,分析表明,支系A的分布曲线呈单峰形,Fs值为-24．6491,P值为0．0000,显著偏离中性,表明山羊支系A曾经历群体扩张;支系B呈近似双峰分布,Fs值为-3．3947,P值为0．0980,中性检验差异不显著,表明山羊支系B没有经历群体扩张,群体大小保持相对稳定。山羊支系B可能起源于中国。     

基于COI基因分析盘羊、塔什库尔干羊的遗传多样性和系统进化

[目的]从线粒体细胞色素C氧化酶亚基Ⅰ(cytochrome c oxidase subumitⅠ,COI)基因水平探讨盘羊对家绵羊有无遗传贡献,了解盘羊和塔什库尔干羊遗传多样性和系统进化。[方法]采用粪便DNA提取法、PCR和测序方法对3只盘羊和68只塔什库尔干羊共71个样品的线粒体COI基因全序列进行分析。[结果]塔什库尔干羊单倍型多样度为0.890±0.001 1,核苷酸多样性为0.00421,表明塔什库尔干羊遗传多样性较为丰富;系统发育分析表明,塔什库尔干羊有4个母系起源,分别是A、B、C和E世系。[结论]欧洲摩弗伦羊对塔什库尔干羊世系B有遗传贡献,基于线粒体COI基因分析,尚未发现盘羊对塔什库尔干羊有遗传贡献的分子证据。     

绵羊线粒体DNA D-loop区串联重复序列变异研究

对来自69个我国地方绵羊品种和8个国外引入品种共计77个个体线粒体DNA控制区长度为75bp的串联重复序列进行了测序分析。在309个重复序列中检测到28个变异位点,其中7个为具有2个变异体的单现突变,1个为具有3个变异体的单现突变,20个为具有2个变异体的简约位点。由28个变异位点中归纳出63个单倍型,其中单倍型Ⅰ和单倍型Ⅲ具有较高的比例,分别为12．94％和30.42％。研究结果揭示我国地方绵羊可能起源于两个母系祖先。哈萨克羊和阿勒泰羊间以及蒙古羊和乌珠穆沁羊间分别具有较近的亲缘关系且没有明显的遗传分化。藏绵羊、蒙古羊和乌珠穆沁羊相对哈萨克羊和阿勒泰羊而言具有较低的遗传多样性。     

绵羊X染色体59571364与59912586位点在脂尾、瘦尾绵羊群体中的多态性检测及分析

为了探讨绵羊X染色体上两处SNP(59571364和59912586)与绵羊尾脂沉积性状的关系, 继而为利用分子标记辅助选择育种技术培育低脂绵羊品种提供依据, 文章以尾型极端差异的阿勒泰羊、湖羊、中国美利奴细毛羊以及萨福克羊为研究对象, 利用PCR-RFLP检测两位点在群体中的多态性, 并分析了两个SNP位点在阿勒泰羊群体中的单倍型。结果表明：59571364位点的TT基因型和59912586位点的GG基因型在瘦尾中国美利奴与萨福克羊群体中属于优势基因型, 而两种基因型在脂尾(臀)阿勒泰羊与湖羊群体中比率均不足2%; 两位点在阿勒泰羊群体中的单倍型分析结果显示, CA单倍型为主单倍型, 比率高达55%, CA与TA两单倍型约占88.33%。以上结果提示, 绵羊X染色体59571364与59912586位点在脂尾(臀)与瘦尾绵羊群体中分布存在较大差异, 可作为理想的分子标记应用于高、低脂绵羊品种选育。     

利用12个血液蛋白和非蛋白标记分析同羊起源及系统地位

采用典型群随机抽样在陕西省白水县抽取同羊样本。采用淀粉凝胶和醋酸纤维薄膜检测12个的结构基因座位的遗传多型性,结果在同羊中发现10个多型座位：运铁蛋白（Tf）、碱性磷酸酶（Alp）、亮氨酸氨肽酶（Lap）、芳基酯酶（Ary-Es）、血红蛋白β（Hb-β）、X-蛋白（X-p）、碳酸酐酶（CA）、过氧化氢酶（Cat）、苹果酸脱氢酶（MDH）和赖氨酸（Ly）;而白蛋白（Al）和后白蛋白（Po）为单态。采用遗传贴近度和系统关系聚类分析两种方法分析同羊起源及系统地位。结果表明两种方法均支持同羊属于蒙古羊系统,同羊起源于蒙古羊,这与同羊的育成史实相符。和聚类分析方法相比,遗传贴近度分析方法可以更为有效地用于中亚以东南绵羊群体的血统判别,可以更有效地反映同羊的育成过程。     

中国部分黄牛品种mtDNA遗传多态性研究

对我国8个黄牛品种22个个体的mtDNA D-loop区910bp全序列进行了分析。结果表明：8个黄牛品种D-loop区序列中,A T平均含量为61．65％;经比对,共检测到66个核苷酸多态位点,约占核苷酸总数的7．25％;D-loop全序列突变类型有5种,即转换、颠换、插入、缺失及转换与颠换共存,它们分别占核苷酸多态位点的81．82％、6．06％、7．57％、3．03％及1．52％。以欧洲牛mtDNA D-loop全序列为标准,8个黄牛群体D-loop的平均核苷酸变异率分3个层次：西镇牛、蒙古牛、黑白花牛及秦川牛的核苷酸变异率最低,分别为0．37％、0．44％、0．52％和0．66％;南阳牛与郏县红牛的核苷酸变异率居中,分别为1．91％和2．02％;晋南牛与岳阳牛的核苷酸变异率最高,分别为4．47％和4．73％。中国黄牛品种内D-loop区序列歧异度为0．55％～5．39％,品种间序列歧异度为1．21％～6．59％。在所测黄牛个体中,mtDNA D-loop序列由19种单倍型组成,单倍型比例为86．36%,说明中国黄牛mtDNA遗传多态性很丰富。由此构建了中国8个黄牛品种的NJ分子系统树,聚类分析表明：所测黄牛的mtDNA D-loop序列表现为3个单倍型组,从而揭示中国黄牛可能有3个母系起源,以普通牛起源和瘤牛起源为主。     

黄河裸裂尻鱼五种群mtDNA控制区的遗传结构

黄河裸裂尻鱼（Schizopygopsis pylzovi）主要分布于青藏高原东北部兰州以上黄河干支流水系和高原北部柴达木内流水系,由于其对高原特殊生境表现出独特的适应性而受研究者关注。本文采用聚合酶链式反应（PCR）和直接测序的方法获得了一个分布于柴达木内流水系种群和四个黄河支流种群共99条个体的线粒体DNA（mtDNA）控制区（D-100p）821bp核苷酸的序列,探讨了其种群遗传结构和分化。99条个体的序列经比对后,发现77个（9．37％）多态性位点,共定义了53个单倍型,其中有一个单倍型（DT12）为两个种群（大通种群和互助种群）所共享,其余52个单倍型均为某个种群所特有。遗传多样性分析表明,柴达木种群的单倍型多样度（h=0．79±0．06）和核苷酸多样度（π=0．0027±0．0017）均略低于黄河支流各种群。AMOVA分析结果显示,遗传差异主要发生在种群之内,占75．39％,而小同水系的种群间只有24．61％,各种群间成对的Fst及遗传距离均表明种群间出现了一定程度的种群分化,但系统进化树最示出一种混杂的单倍型分布格局,提示青藏高原东北部地质事件所造成的水系间的隔离形成较晚。单倍型歧点分布呈现为单峰以及中性检验Tajima的D（-1．497,P=0．058）和Fu的Fs（-24．741,P=0．001）结果综合表明,黄河裸裂尻鱼在青藏高原隆升过程中曾经历过近期的种群扩张事件[动物学报54（6）：972—980,2008]。     

中国主要鹅品种的线粒体DNA多态性与起源分化研究

运用19种限制性内切酶对中国11个家鹅品种138个样本进行了mtDNA的限制性片段长度多态性（RFLP）分析。在使用的19种内切酶中,有7个酶检测出多态。综合27种限制性态型（restrictionmorph）,可得到6种mtDNA单倍型（hopotype）。伊犁鹅与另外10个鹅品种没有共享的单倍型,遗传距离和UPGMA聚类分析也表明,伊犁鹅与这些品种具有不同的起源。EcoRV、HaeⅡ、HincⅡ和KpnⅠ4种酶的限制性态型可作为鉴别两种起源家鹅的母系遗传标记。起源于鸿雁的10个鹅品种群体内出现一定的遗传差异,其群体多态度（π）、单倍型间平均遗传距离（P）、品种间平均净遗传距离（δnet）分别为0．025％、0．266％和0．029％。白羽鹅品种在形成过程中经历过创立者效应（foUndereffect）。这10个鹅品种可能起源于两个不同地理区的鸿雁类群。     

白蚁与动物及微生物的共生类型及其机制

综述了白蚁螱客的主要种类、共生关系及相关机制的研究进展。白蚁螱客中,已报道的动物种类达170种。在与动物的共生关系中存在偏利共生（宾主共栖和异种共栖）、互利共生和无关共生三种;在与微生物的共生关系中,存在与内生菌（原生动物、细菌、真菌和放线菌）和外生菌（蚁巢伞菌等）间的互利关系。指出了白蚁与螱客研究中存在的问题,给出了解决方案,并提出了今后可能的研究热点或方向,为白蚁的综合利用（如纤维素酶）及今后研究物种间的协同进化提供了基础资料。     

New amino-dithiaphospholanes and phosphoramidodithioites and their rhodium and iridium complexes

New sulfur derivatives of phosphoramidite ligands were synthesized and the impact of the sulfur unit on the spectroscopic properties of their rhodium and iridium complexes was investigated. The new ligands Bn₂NPSCH₂CH₂S^a(P-S^a) (Bn = benzyl, 4), Bn₂NPSCHCHS^a(CH₂)₃C^aH₂(P-S^a)(C^a-S^a) (6) and Bn₂NP(4-XC₆H₄OMe)₂ (X = S, 7a; X = O, 7b) were converted to the rhodium and iridium complexes trans-[Rh(CO)Cl(L)₂] (L = 4, 6, 7), [RhCl(COD)(L)] (L = 4, 6, 7), [IrCl(COD)(7a)] and [IrCl₂Cp∗(6)]. For comparison, some phosphoramidite complexes of these formulations also were synthesized. The new metal complexes were spectroscopically analyzed. For the carbonyl complexes, the ν_CO IR stretching frequencies were lower than for the corresponding phosphite and phosphoramidite ligands. The ¹J_PRh coupling constants for the rhodium complexes with the new ligands were also smaller than for the respective phosphoramidite and phosphite complexes. Finally, the ¹J_PSe coupling constants of the selenides of the new ligands were lower than those of the phosphoramidite ligands but higher than for PPh₃. The spectroscopic data reveal that the new thio ligands 4, 6 and 7a are more electron donating than phosphites and phosphoramidites but less electron donating than PPh₃.     

Astrocytes and Synaptosomes Transport and Metabolize Lactate and Acetate Differently

Astrocytes transport the monocarboxylate acetate, but synaptosomes do not. The reason for this is unknown, because both preparations express monocarboxylate transporters (MCT). The transport and metabolism of lactate, another monocarboxylate, was examined in these two preparations, and the results were compared to those for acetate. Lactate transport is more rapid in astrocytes than in synaptosomes, but of lower affinity (Kms of 17 and 4 mM, respectively). Lactate (0.2 mM) is metabolized to CO2 more rapidly in synaptosomes than in astrocytes (rates of 0.37 and 0.07 nmol x mg protein(-1) x min(-1), respectively). The reason for this is unclear, but cellular differences in lactate dehydrogenase isotype expression may be involved. Acetate is metabolized to CO2 more rapidly in astrocytes than in synaptosomes (rates of 0.43 and 0.02 nmol x mg protein(-1) x min(-1), respectively). This is likely due to cellular differences in the expression of monocarboxylate transporter subtypes.     

Rapporteur report: Basics and technology and metrology and standards

The first and second sessions of the Workshop focussed on the basics of ultrasound and infrasound, their applications in both industry and medicine, and metrology and protection standards for ultrasound applications.     

Relative and combined effects of ethanol and protein deficiency on strontium and barium bone content and fecal and urinary excretion

To elucidate accumulation of minerals in human iliac arteries with aging, the content of minerals was analyzed by inductively coupled plasma atomic emission spectrometry. Bilateral common, internal, and external iliac arteries of 16 men and 8 women, ranging ages from 65 to 93 yr, were examined. It was found that an extremely high accumulation of calcium and phosphorus occurred in the common iliac artery at old age, being higher than that of the internal and external iliac arteries. It should be noted that the accumulation of calcium and phosphorus is the highest in the common iliac artery among the human arteries examined to date. Regarding sexual differences, the content of calcium and phosphorus in the common and internal iliac arteries was higher in women than in men, whereas their content in the external iliac artery was lower in women than in men.     

Cytoskeleton and mitochondrial morphology and function

It has been well established that the cytoskeleton is an essential modulator of cell morphology and motility, intracytoplasmic transport and mitosis, however cytoskeletal linkage to the organelles has not been unequivocally demonstrated. Indeed, cytoskeleton appears to be essential in determining and modulating gene phenotype as a function of cellular environment. According to recent studies, the organization of the cytoskeleton network together with associated protein(s) could be essential in regulating mitochondrial function and particularly the permeability of the mitochondrial outer membrane to ADP. The aim of this chapter is to summarize the main properties of the cytoskeletal environment of mitochondria and the possible role(s) of this network in mitochondrial function in myocytes.     

Cross-reactivity between storage and dust mites and between mites and shrimp

Many patients have sensitivities to multiple species of storage and house dust mites. It is not clear if this is because patients have multiple sensitivities to species-specific mite allergens or if these mites share many cross-reacting allergens. Our objective was to further define the cross-allergenicity between several species of storage and house dust mites using crossed-immunoelectrophoresis (CIE), crossed-radioimmunoelectrophoresis (CRIE), immunoblotting, and ELISA. CIE and CRIE reactions revealed that storage mites shared two cross-antigenic molecules and one of these bound IgE in a serum pool from mite allergic patients. Antibody in anti-sera built to each species of mite recognized many SDS–PAGE resolved proteins of other mite species and this suggested the potential for other cross-reactive allergens. Among patient sera, IgE bound to many different proteins but few had IgE that bound to a protein with common molecular weights across the mite species and this suggested mostly species-specific allergens. Antiserum built to each mite species precipitated one protein in shrimp extracts that bound anti-Der p 10 (tropomyosin) and IgE in the serum pool. Anti-Der p 10 showed strong binding to shrimp tropomyosin but very little to any of the mite proteins. ELISA showed the mite extracts contained very little tropomyosin. The storage and dust mites investigated contain mostly species-specific allergens and very small amounts of the pan-allergen tropomyosin compared to shrimp and snail.     

人血管能抑素基因的克隆、表达及其表达产物的纯化和生物活性测定

以人胎盘脐带组织为材料,提取组织总RNA,用netRTPCR方法合成人血管能抑素cDNA基因,将该cDNA克隆进pSP72载体获得重组质粒pSP72C, DNA序列分析结果与预期序列一致。用BamHⅠ和NdeⅠ双酶切,切下pSP72C上的血管能抑素cDNA,插入pET3c载体的相应位点获得重组表达质粒pETC, 转化E. coli BL21(DE3), SDSPAGE分析显示：在IPTG诱导下,血管能抑素基因获得了高效表达,表达量约占菌体总蛋白的 27.9 %,主要以包涵体形式存在。包涵体经过洗涤、裂解、蛋白复性以及Sephadex G75凝胶过滤层析等步骤后,获得了纯度达91.4 %的人血管能抑素。CAM实验证明10 μg纯化蛋白就能显著抑制鸡胚新生血管生成。     

Vascular volumes and hematology in male and female runners and cyclists

To examine the hypothesis that foot-strike hemolysis alters vascular volumes and selected hematological properties is trained athletes, we have measured total blood volume (TBV), red cell volume (RCV) and plasma volume (PV) in cyclists (n = 21) and runners (n = 17) and compared them to those of untrained controls (n = 20). TBV (ml x kg(-1)) was calculated as the sum of RCV (ml x kg(-1)) and PV (ml x kg(-1)) obtained using 51Cr and 125I-labelled albumin, respectively. Hematological assessment was carried out using a Coulter counter. Peak aerobic power (VO2peak) was measured during progressive exercise to fatigue using both cycle and treadmill ergometry. RCV was 15% higher (P < 0.05) in male cyclists [35.4 (1.0), mean (SE); n = 12] and runners [35.3 (0.98); n = 9] compared to the controls [30.7 (0.92); n = 12]. Similar differences existed between the female cyclists [28.2 (2.1); n = 9] and runners [28.4 (1.0); n = 8] compared to the untrained controls [24.9 (1.4); n = 8]. For the male athletes, PV was between 19% (cyclists) and 28% (runners) higher (P < 0.05) in the trained athletes compared to the untrained controls. The differences in PV between the female groups were not significant. Although the males had a higher (P < 0.05) TBV, RCV and PV than the females, no differences between cyclists and runners were found for either gender. Mean cell volume was not different between the athletic groups. VO2peak (ml x kg(-1) x min(-1)) was higher (P < 0.05) in both male [68.4 (1.5)] and female [54.8 (2.1)] runners when compared to the untrained males [47.1 (1.0)] and females [40.5 (2.1)]. Although differences existed between the genders in VO2peak for both cyclists and runners, no differences were found between the athletic groups within a gender. Since the vascular volumes were not different between cyclists and runners for either the males or females, foot-strike hemolysis would not appear to have an effect on that parameter. The significant correlations (P < 0.05) found between VO2peak and RCV (r = 0.64 and 0.64) and TBV (r = 0.82 and 0.63) for the males and females, respectively, suggests a role for the vascular system in realizing a high aerobic power.