The successful double cryopreservation of rabbit (Oryctolagus cuniculus) semen in large volume using the directional freezing technique with reduced concentration of cryoprotectant

Using directional freezing, Our objective was to cryopreserve rabbit semen and achieve fertility that was equal or higher than that achieved with conventional freezing. The working hypothesis was that controlling the ice-front propagation would allow redu

Genetic diversity of Chinese domestic goat based on the mitochondrial DNA sequence variation

The aim of this study was to characterize the genetic diversity of domestic goat in China. For this purpose, we determined the sequence of the mitochondrial DNA (mtDNA) control region in 72 individuals of the Yangtze River delta white goat, and reanalysed

Study on the variation of chemical constituents during combination of ginseng with trogopteroum feces and semen raphani by high performance liquid chromatography-mass spectrometry

A high performance liquid chroatography-electrospray ionization-mass spectrometric method was developed for analysis and identification of ginsenosides from the decoction of ginseng, ginseng with trogopteroum feces and ginseng with semen raphani. Ten ginsenosides were separated and detected. The content variation of these ginsenosides was researched. The experimental results showed, that ginsenosides were less in compatible decoction than in separate one expect Ro. the stripping of ginsenosides were restrained by semen raphani and during combination of ginseng with trogopteroum feces, the precipitates were produced by ginsenosides.

剪切流动法研究表面固定化配基与目标分子的相互作用力

基于剪切流动腔技术,以微球作为受力载体,设计了一套可用于研究表面固定化配基与目标分子特异性相互作用力的实验和分析方法,并以人免疫球蛋白G (human IgG)和羊抗人免疫球蛋白G(goat anti-human IgG)分别作为模型配基和模型目标分子进行了研究.基于平面Poiseuille层流模型设计了流场参数,以数值计算结果验证了设计的合理性.使用牛血清白蛋白(BSA)作为非特异性对照,判断微球与基片表面的结合力来自配基和目标分子的生物特异性相互作用,并由进一步的目标分子灭活对比实验确认了这一结论.实验观察到微球与基片表面的结合力受到配基面密度的影响,说明发生结合的是多对而非单对蛋白质分子.将95%的微球被剥离时对应的壁面剪切率设定为临界剪切率,由大量实验结果拟合得到了临界剪切率与配基面密度间的定量关系.在受力分析模型中,考虑到多分子的结合,以及分子键位置不同造成的力臂长度的差异,最终计算得到单对配基与目标分子的平均结合力约为342pN.

直墙前破波波浪力──对修正过的Gota方法的探讨

对被重新修正后的Goat在墙波压力计算方法进行了分析，主要是对其无因次波浪力随基床肩宽和相对波高的变化进行分析，结果发现，Goat法计算的无因次波浪力随基床肩宽的变化，有一最不利肩宽，此不利肩宽与相对波高无关；在同一水深条件下，Goat法计算的无因次波浪力随相对波高的增大而增大。此两点与试验数据不符。

Study of Interaction Force between Antigen and Antibody Using Flow Chamber Method

A parallel plate flow chamber was used to study the interaction force between human IgG (immobilized on a chip surface as ligand) and goat anti-human IgG (immobilized on microspheres surface as receptor). First, it was demonstrated that the binding force between the microspheres and the chip surface came from the bio-specific interaction between the antigen and the antibody. Secondly, it was obtained that the critical shear rate to detach microspheres from the chip surface increases with the ligand surface concentration. Finally, two models to estimate the antigen-antibody bond strength considering bonds' positions were proposed and analyzed.

Phylogenomic study of the subfamily Caprinae by cross-species chromosome painting with Chinese muntjac paints

Chromosomal homologies have been established between the Chinese muntjac (Muntiacus reevesi, MRE, 2n = 46) and five ovine species: wild goat (Capra aegagrus, CAE, 2n = 60), argall (Ovis ammon, OAM, 2n = 56), snow sheep (Ovis nivicola, ONI, 2n = 52), red goral (Naemorhedus cranbrooki, NCR, 2n = 56) and Sumatra serow (Capricornis sumatraensis, CSU, 2n = 48) by chromosome painting with a set of chromosome-specific probes of the Chinese muntjac. In total, twenty-two Chinese muntjac autosomal painting probes detected thirty-five homologous segments in the genome of each species. The chromosome X probe hybridized to the whole X chromosomes of all ovine species while the chromosome Y probe gave no signal. Our results demonstrate that almost all homologous segments defined by comparative painting show a high degree of conservation in G-banding patterns and that each speciation event is accompanied by specific chromosomal rearrangements. The combined analysis of our results and previous cytogenetic and molecular systematic results enables us to map the chromosomal rearrangements onto a phylogenetic tree, thus providing new insights into the karyotypic evolution of these species.

山羊mtDNA 多态性及其起源分化研究

　本文用14 种识别6 碱基的限制性内切酶Apa Ⅰ、BamH Ⅰ、Bgl Ⅰ、Bgl Ⅱ、Dra Ⅰ、 EcoR Ⅰ、EcoR Ⅴ、Kpn Ⅰ、Pvu Ⅱ、Pst Ⅰ、Sac Ⅰ、Sal Ⅰ、Sma Ⅰ和Xho Ⅰ研究了来自我国12 个省 和自治区共计18 个地方山羊品种218 个个体mtDNA 的RFL P ,并运用Nei 氏公式计算了各限制 性类型间的遗传距离P 和群体遗传多态度π值。结果表明,在研究的所有个体中共检测到41 个 酶切位点,18 种限制性态型,其中BamH Ⅰ、Bgl Ⅱ、Dra Ⅰ、EcoR Ⅰ和Sal Ⅰ共5 种酶表现出多态。 18 种限制性态型可归结为6 种基因单倍型,单倍型Ⅰ(BamH Ⅰ2B、Bgl Ⅱ2A、Dra Ⅰ2A、EcoR Ⅰ2A 和Sal Ⅰ2A) 和单倍型Ⅱ(BamH Ⅰ2B、Bgl Ⅱ2A、Dra Ⅰ2A、EcoR Ⅰ2A 和Sal Ⅰ2B) 为两种基本单倍 型,研究结果提示我国地方山羊品种起源于两种不同的母系祖先。各限制性类型间的平均遗传距 离为0100436 ,整个群体的平均遗传多态度π值为010487 % ,表明我国地方山羊品种mtDNA 遗传 多样性比较贫乏,分化程度较低。

黔东南小香羊与贵州原有其它山羊品种的线粒体DNA 多态性比较

用B am H É、B g lÉ、B g lÊ、D raÉ、E coR É、E coR Í、H indË、Kp nÉ、 P stÉ、P vuÊ、S acÉ、S a lÉ、Sm aÉ、S tuÉ、X hoÉ 15 种识别6 碱基的限制性内 切酶对黔东南小香羊和贵州原有3 个地方山羊品种的93 只个体的m tDNA 进行分 析表明, B am H É、H indË 和S a lÉ 3 种酶表现多态性; 共检出18 种限制性多态型, 归纳得到3 种单倍型, 以单倍型É 和Ê 为基本单倍型。根据此2 种基本单倍型在所 比较各品种中的不同分布比例, 以及遗传距离分析和品种间的聚类关系, 表明黔东 南小香羊的群体遗传构成与贵州省原有其它3 个山羊品种不同, 从而为进一步确认 其为一独立的品种提供了必要的分子生物学依据。

Mitochondrial diversity and phylogeographic structure of Chinese domestic goats

China has numerous native domestic goat breeds, but so far there has been no extensive study on genetic diversity, population demographic history, and origin of Chinese goats. Here, we examined the genetic diversity and phylogeographic structure of Chinese domestic goats by determining a 481-bp fragment of the first hypervariable region of mitochondrial DNA (mtDNA) control region from 368 individuals representing 18 indigenous breeds. Phylogenetic analyses revealed that there were four mtDNA lineages (A-D) identified in Chinese goats, in which lineage A was predominant, lineage B was moderate, and lineages C and D were at low frequency. These results further support the multiple maternal origins of domestic goats. The pattern of genetic variation in goat mtDNA sequences indicated that the two larger lineages A and B had undergone population expansion events. In a combined analysis of previously reported sequences and our sequences belonging to lineage B, we detected two subclades, in which one was unique to eastern Asia and another was shared between eastern and southern Asia. A larger genetic variation in eastern Asia than southern Asia and the pattern of phylogeographic variation in lineage B suggest that at least one subclade of lineage B originated from eastern Asia. There was no significant geographical structuring in Chinese goat populations, which suggested that there existed strong gene flow among goat populations caused by extensive transportation of goats in history. (c) 2005 Elsevier Inc. All rights reserved.

云南保种山羊线粒体DNA限制性酶切研究

　采用碱变性法,提取来自云南省不同地区4个保种山羊的13个个体的线粒体DNA(mtDNA),并用ApaⅠ,AvaⅠ,BamHⅠ,BclⅠ,BcIⅠ，BglⅡ,ClaⅠ,DraⅠ, EcoRⅠ,EcoRⅤ,HaeⅠ,HindⅢ,KpnⅠ,PstⅠ,PvuⅡ,SacⅠ,SalⅠ,SmaⅠ,StuⅠ和XhoⅠ等20种限制性内切酶进行酶切分析。结果发现它们的线粒体DNA的分子量大 小约为15.8Kb；不同限制性内切酶的酶切位点分别为：DraⅠ有7个酶切位点，AvaⅢ有6个酶切位点，EcoRⅤ和StuⅠ共有5个酶切位点，HindⅡ和HaeⅡ有4个酶 切位点，BamHⅠ,BglⅡ,PstⅠ和PvuⅡ有3个酶切位点，ApaⅠ，ClaⅠ有两个酶切位点，其余有1个酶切位点。各保种山羊间未发现变异，说明云南的4个保种山 羊极可能来自于共同的母性祖先。

Cryopreservation of cynomolgus monkey (Macaca fascicularis) spermatozoa in a chemically defined extender

Aim: To establish a method for cynomolgus monkey sperm cryopreservation in a chemically defined extender. Methods: Semen samples were collected by electro-ejaculation from four sexually mature male cynomolgus monkeys. The spermatozoa were frozen in straws by liquid nitrogen vapor using egg-yolk-free Tes-Tris (mTTE) synthetic extender and glycerol as cryoprotectant. The effects of glycerol concentration (1%,3%, 5%, 10% and 15% [v/v]) and its equilibration time (10 min, 30 min, 60 min and 90 min) on post-thaw spermatozoa were examined by sperm motility and sperm head membrane integrity. Results: The post-thaw motility and head membrane integrity of spermatozoa were significantly higher (P < 0.05) for 5% glycerol (42.95 +/- 2.55 and 50.39 +/- 2.42, respectively) than those of the other groups (1%: 19.19 +/- 3.22 and 24.84 +/- 3.64; 3%: 34.23 +/- 3.43 and 41.37 +/- 3.42; 10%: 15.68 +/- 2.36 and 21.39 +/- 3.14; 15%: 7.47 +/- 1.44 and 12.90 +/- 2.18). The parameters for 30 min equilibration (42.95 2.55 and 50.39 2.42) were better (P < 0.05) than those of the other groups (10 min: 31.33 +/- 3.06 and 38. 98 +/- 3.31; 60 min: 32.49 +/- 3.86 and 40.01 +/- 4.18; 90 min: 31.16 +/- 3.66 and 38.30 +/- 3.78). Five percent glycerol and 30 min equilibration yielded the highest post-thaw sperm motility and head membrane integrity. Conclusion: Cynomolgus monkey spermatozoa can be successfully cryopreserved in a chemically defined extender, which is related to the concentration and the equilibration time of glycerol.

不同世代改良型路南奶山羊血液免疫学参数的比较研究

本文测定了“改良型路南奶山羊”的亲本及改良不同世代 (F1,F2 ,F3,F4 )母羊的外周血T淋巴细胞转化率、ANAE阳性率、B淋巴细胞ZYC花环率、红细胞IC花环率、C3b花环率以及血清γ -球蛋白等免疫学参数 ,比较研究其随着世代增递的变异情况。结果表明 :从亲代到F4代随着改良世代的递增 ,T淋巴细胞转化率、ANAE阳性率、红细胞C3b花环率和IC花环率和血清γ -球蛋白含量均呈F1>F2 >P >F3>F4变化 ,ZYC花环率呈F1>P >F2 >F3>F4变化。回归分析的结果表明 :从亲代到改良四代 ,上述各项免疫学参数均呈显著负相关 ,趋势线为下降趋势。即随着世代的递增有逐代递减的趋势。结果提示 :用西农莎能羊改良路南圭山羊 ,其改良一代和二代的上述各项血液免疫学参数均略高于路南圭山羊 ,以后随着世代的递增有逐代递减的趋势。预示着随着世代的递增 ,改良型路南奶山羊的体质状况 ,免疫功能和抗病能力可能会出现下降趋势。