Monitoring the emission of volatile organic compounds from the leaves of Calocedrus macrolepis var. formosana using solid-phase micro-extraction

In this study, solid-phase micro-extraction (SPME) fibers coated with polydimethylsiloxane/divinylbenzene (PDMS/DVB), coupled with gas chromatography/mass spectrometry, were used to monitor the emission patterns of biogenic volatile organic compounds (BVOCs) from leaves of Calocedrus macrolepis var. formosana Florin. in situ. In both sunny and rainy weather, the circadian profile for BVOCs from C. macrolepis var. formosana leaves has three maximum emission cycles each day. This kind of emission pattern might result from the plant’s circadian clock, which determines the rhythm of terpenoid emission. Furthermore, emission results from the leaves demonstrated that the circadian profile of α-pinene observed was opposite to the profiles of limonene and myrcene, a difference that may be attributable to two different subpathways for terpenoid biosynthesis.     

Assessment of the tumorigenesis and drug susceptibility of three new canine mammary tumor cell lines

Three canine mammary tumor (CMT) cell lines, namely DE-E, DE-F and DE-SF, have been established from a surgically excised specimen of a malignant mammary tumor. These CMT cell lines have been cultured for over 200 passages. The cell doubling time was estimated to be approximately 30 h for all three cell lines. DE-E, DE-F and DE-SF were epithelial, fibroblast and spindle fibroblast in morphology, respectively. Under electron microscope, DE-F and DE-SF cells displayed a higher nucleus/cytoplasm ratio as compared with DE-E. Variation in chromosome number was also observed in the three cell lines. In addition to the morphological characteristics, these cell lines displayed differential patterns of several known mammary tumor cell markers. Following xenotransplantation of the CMT cells into nude mice, DE-F and DE-SF developed tumors within 2 weeks, whereas DE-E failed to develop any visible tumor up to 8 weeks after injection. Lastly, the CMT cell lines exhibited differential chemoresistance to several anti-tumor drugs, including melatonin, cyclosporine A, tamoxifen and indole, suggesting that these cell lines can be used as a comparative experimental model for the tumorigenesis of mammary carcinomas and a valuable tool for anti-cancer drug screening.     

The phylogeographic system survey of native sheep breeds in the eastern and southern Central Asia

The genetic diversity and phylogenetic survey of native sheep breeds in the eastern and southern Central Asia were assessed in the present study. The clustering, principal components, structure and F statistics all demonstrate that the native sheep breeds in these regions be classified into two genetic groups: Mongolia‐Tibetan sheep group and South‐Southeast Asia sheep group. The Mongolia sheep group and the Tibetan sheep group had a certain degree of gene communication from the ancient times. In the present study we demonstrated that the Chinese native sheep populations belonged to Mongolia‐Tibetan sheep group. However, the relationships among the sheep populations in Mongolia sheep group in China were not closely related to the geographical distance among sheep populations.     

Dog erythrocyte antigens 1.1, 1.2, 3, 4, 7, and Dal blood typing and cross‐matching by gel column technique

Background: Testing for canine blood types other than dog erythrocyte antigen 1.1 (DEA 1.1) is controversial and complicated by reagent availability and methodology. Objectives: The objectives of this study were to use available gel column technology to develop an extended blood‐typing method using polyclonal reagents for DEA 1.1, 1.2, 3, 4, 7, and Dal and to assess the use of gel columns for cross‐matching. Methods: Dogs (43–75) were typed for DEA 1.1, 1.2, 3, 4, 7, and Dal. Methods included tube agglutination (Tube) using polyclonal reagents, a commercially available DEA 1.1 gel column test kit (Standard‐Gel) using monoclonal reagent, and multiple gel columns (Extended‐Gel) using polyclonal reagents. Blood from 10 recipient and 15 donor dogs was typed as described above and cross‐matched using the gel column technique. Results: Of 43 dogs typed for DEA 1.1, 23, 25, and 20 dogs were positive using Standard‐Gel, Extended‐Gel, and Tube, respectively. Typing for DEA 1.2 was not achievable with Extended‐Gel. For 75 dogs typed for DEA 3, 4, and 7, concordance of Extended‐Gel with Tube was 94.7%, 100%, and 84%, respectively. Dal, determined only by Extended‐Gel, was positive for all dogs. Post‐transfusion major cross‐matches were incompatible in 10 of 14 pairings, but none were associated with demonstrable blood type incompatibilities. Conclusions: Gel column methodology can be adapted for use with polyclonal reagents for detecting DEA 1.1, 3, 4, 7, and Dal. Agglutination reactions are similar between Extended‐Gel and Tube, but are more easily interpreted with Extended‐Gel. When using gel columns for cross‐matching, incompatible blood cross‐matches can be detected following sensitization by transfusion, although in this study incompatibilities associated with any tested DEA or Dal antigens were not found.     

Calcium homeostasis and its relationship to superoxide production in blood and milk neutrophils of lactating goats

Polymorphonuclear neutrophils (PMN), which comprise over 70% of the somatic cells in goat milk, are a major cellular component of innate immunity in the goat mammary gland. However, the function of milk PMNs is modified after diapedesis compared to PMNs in blood. As many aspects of PMN activity depend directly on intracellular Ca²⁺ concentration ((Ca²⁺)_i), the present study aimed to determine the changes in Ca²⁺ homeostasis of milk PMNs from lactating goats compared to autologous blood PMNs, and to examine the significance of these variations to the immuno-competency of milk PMNs. The intracellular Ca²⁺ store of freshly prepared milk cells was estimated from the elevation of (Ca²⁺)_i after ionomycin treatment, which was found to be significantly less than blood PMNs. Replenishment of the intracellular Ca²⁺ store in milk cells after intracellular Ca²⁺ depletion by Bapta-AM followed by spiking with 2.5 mM Ca²⁺ for 20 min was also compared to that of blood PMNs, showing that after depletion/spiking the intracellular Ca²⁺ store in milk cells was much less than blood PMNs. The production of superoxide anion (O₂^?) in vitro in response to (Ca²⁺)_i-dependent or (Ca²⁺)_i-independent modulators was used to evaluate the relevance of altered Ca²⁺ homeostasis on the immuno-competency of milk cells compared to blood PMNs. The results indicated that milk cells produced similarly low levels of O₂^? as blood PMNs when treated with ionomycin. However, the amount of O₂^? produced by milk cells in response to phorbol 12-myristate 13-acetate (PMA) stimulation, although greater than ionomycin treatment, was significantly less than that of blood PMNs. The capacity for O₂^? production by both cell types in response to PMA reverted to the resting state with use of the protein kinase C (PKC) inhibitor, staurosporine. In conclusion, the current study demonstrated an irreversible shortage of intracellular Ca²⁺ in the milk PMNs of lactating goats compared to blood PMNs. It also showed that preliminary O₂^–production, primed by ionomycin treatment, remained unchanged in milk PMNs, despite the shortage in intracellular Ca²⁺, but decreased O₂^? production capacity, mediated via the PKC pathway, in milk PMN. It is suggested that the defects in Ca²⁺ homeostasis in milk PMNs of lactating goats is partially attributable for the post-diapedesis functionality modifications.     

猪圆环病毒病和猪繁殖与呼吸综合征二联灭活疫苗免疫效力

利用猪圆环病毒病(PCVD)和猪繁殖与呼吸综合征(PRRS)二联灭活疫苗免疫4周龄仔猪,免疫后7、14、21、28d采血,测定免疫后血清中抗PRRSV、PCV中和抗体和ELISA抗体较价,并于免疫后28d分别用PRRSV和PCV2攻毒,攻毒后每日检测猪体温和观察临床症状,于攻毒后21d所有存活猪全部扑杀,分别对每头猪肺、肝、脾、淋巴结、心和血进行剖解病理和组织病理学观察及PRRSV和PCV2核酸检测,以确定该疫苗免疫保护率。结果表明,所有试验猪于免疫后7d血清抗体开始检出,28d达到峰值,免疫后28dPRRSV和PCV2攻毒保护率均为100%(5/5)。     

VOD网络自学系统应用研究

流媒体技术目前已经相当成熟,而且应用也很广泛,本文主要阐述了如何利用流媒体技术,选择合适的方案,结合学校机房现有资源的特点来搭建高效流畅的网络视频教学点播系统,从而给学生提供一个自助学习的理想环境。     

新麦草(Psathyrostachys juncea)两个高分子量谷蛋白亚基基因上游序列的克隆与分析

应用PCR技术对新麦草PI429801和PI406469的高分子量谷蛋白基因上游序列进行了克隆,得到长度分别为998和1000 bp的HMW-N1和HMW-N2序列。比较发现,HMW-N1与麦类HMW-GS基因上游序列一致性在85%以上,与滨麦(Leymus mollis)序列DQ073542一致性最高,达98%;HMW-N2与麦类HMW-GS基因上游序列一致性在81%以上,与二角山羊草(Aegilops bicornis)序列AY611726一致性最高,达98%。系统进化分析表明,HMW-N1与含Ns染色体组的赖草属物种的同源基因序列DQ073551、FJ600498和DQ073546亲缘关系最近,HMW-N2却与含D染色体组节节麦和普通小麦的同源基因序列FJ008134、AY248704和DQ537337以及含S染色体组的山羊草物种的同源基因序列AY611726、AY611721和AY611724的亲缘关系较近。同时,HMW-N1和HMW-N2也具有E-box、N-box、partial Enhancer、complete Enhancer、TATA-box和Start等高分子量谷蛋白基因启动子区域的典型特征。本研究结果可为新麦草高分子量谷蛋白基因的克隆提供帮助,对从新麦草属以及其他小麦近缘属物种中分离未知高分子量谷蛋白基因有参考价值和借鉴意义。     

杜鹃花属植物SRAP-PCR反应体系的优化及其应用

采用改良CTAB法提取杜鹃花属植物基因组DNA,对影响SRAP-PCR反应体系中的Mg2+、dNTPs和引物浓度进行优化,建立了杜鹃花属植物SRAP分子标记的扩增体系:20μl的PCR体系中含有模板DNA50 ng、10×PCR buffer(不含Mg2+)、Mg2+2.50 mmol/L、dNTPs 0.20 mmol/L、引物0.40μmol/L、TaqDNA聚合酶1.0 U。并对10种杜鹃花属植物群体进行扩增验证,共获得261条扩增条带,251个多态性位点,多态性位点比率为96.17%,结果表明供试材料间差异明显、多态性较高,该体系适合杜鹃花属植物种间差异性分析,为今后杜鹃花属植物分子生物学的深入研究奠定了基础。