四川地区藏猪戊型肝炎病毒的检测和遗传演化分析

旨在调查四川地区藏猪戊型肝炎病毒（hepatitis E virus，HEV）的流行情况和遗传演化，作者用RT-PCR法对2018—2020年采集自四川省甘孜藏族自治州和阿坝藏族自治州的22个藏猪场332份藏猪粪便样本进行HEV检测，并对阳性样本进行基因分型。RT-PCR检测结果表明HEV核酸阳性率为20.48%（68/332，95% CI=16.3%~25.2%），22个规模藏猪场中HEV猪场阳性率为77.27%（17/22，95% CI=54.6%~92.2%），健康样本阳性率为2.86%（3/105，95% CI=0.6%~8.1%），腹泻样本阳性率为28.63%（65/227，95% CI=22.8%~35.0%），系统进化分析显示所有阳性株均为G4型。为了进一步研究该地区流行毒株的演化过程，以贝叶斯进化分析软件进行分歧时间估算，结果表明，藏猪流行毒株的分歧时间最早为1999年，最晚为2016年。并分别从每年的样本中挑选1份阳性样本得到全基因组序列，核酸序列相似性为89.5%~93.1%。通过对3条全基因组重组分析发现SWU/301/2019存在重组现象，SWU/301/2019重组区域位于ORF1的3 746—4 655 bp。本研究首次对四川地区藏猪源HEV进行调查，结果表明四川藏猪中存在一定程度的HEV感染，为四川地区对藏猪感染HEV的防治以及深入研究四川藏猪源HEV遗传变异及生物学特性提供了参考依据。     

青藏高原地区积雪变化及特征分析

利用我国高原地区1963-2002年的逐月积雪日数资料,以及1972-2011年的格点积雪月频率资料,分析、对比了两种资料在该地区的时空演变特征。结果表明：同时段的(1972-2002)站点资料与格点资料相比,空间分布其高值区与积雪日数增值区均偏西北,偏北程度大致为3~4°,偏西程度大致为2~3°,且中心更为明显。在青藏高原西侧(85°E以西),积雪日数呈整体下降趋势。两种资料的积雪日数EOF分析存在一定的偏差,这与其对应的空间分布差异及气候趋势空间分布存在的偏差相吻合。     

青稞类钙调素蛋白基因CML19的克隆、序列分析及原核表达

根据鉴定得到青稞类钙调素蛋白基因CML19的序列设计引物,以青稞叶片的cDNA为模板,扩增得到目的片段,对目的基因序列进行鉴定和分析,进一步构建该基因的原核表达载体pET28a-CML19,转入大肠杆菌BL21(DE3),利用IPTG诱导外源蛋白质表达并检测。结果表明,扩增得到CML19的CDS序列全长为447bp,编码的蛋白质含有148个氨基酸,分子质量为16.46ku,理论等电点(pI)为4.40,总平均亲水性(GRAVY)为-0.176,不稳定系数为49.59,存在典型的EF手结构域;系统进化分析表明,青稞CML19与山羊草具有较近的亲缘关系;原核表达蛋白结果显示,重组蛋白主要以包涵体形式存在于沉淀中。初步阐明青稞CML19基因的序列特征,为进一步制备抗体、探讨CML19基因的功能奠定基础。     

施氮量对一年生黑麦草光合特性的影响

为了探究不同施氮量对一年生黑麦草(Lolium multiflorum Lamk.)光合特性的影响。本研究以‘特高’宽叶型一年生黑麦草为供试材料,采用盆栽试验,设置了4个处理组,分别为:CK对照组(不施肥)、C1低氮处理组(施鸡粪0.38 kg/m^2,即氮100 kg/hm^2)、C2中氮处理组(施鸡粪0.76 kg/m^2,即氮200 kg/hm^2)、C3高氮处理组(施鸡粪1.34 kg/m^2,即氮350 kg/hm^2)。利用Li-6400便携式光合仪测定开花期一年生黑麦草叶片的主要光合参数和光合特性的变化。结果表明:200 kg/hm^2施氮水平的表观量子效率(apparent quantum yield,AQY)和光饱和点(light saturation point,LSP)最高(分别为12.80 μmol·m^-2·s^-1和1 774.47 μmol·m^-2·s^-1),光能利用区间最大。350 kg/hm^2施氮水平下气孔导度(stomatal conductance,Gs)以及蒸腾速率(transpiration rate,Tr)显著低于其它处理,水分利用力(water use efficiency,WUE)强,但光呼吸速率(photorespiratory rate,Pr)较高,不利于干物质积累。同时,0?200 kg/hm^2施氮范围内,随着光照强度和CO2浓度的增加,各处理的净光合速率(net photosynthetic rate,Pn)均随之增加,但350 kg/hm^2施氮水平下叶片净光合速率的增加反而低于200 kg/hm^2施氮水平。合理的施氮量对黑麦草的光响应和CO2浓度响应曲线有明显的调节作用。在本试验条件下,施氮量为200 kg/hm^2最佳。     

Inductive expression of the NOD1 signalling pathway in chickens infected with Salmonella pullorum

1. The aim of this study was to describe the role of Nucleotide-binding oligomerization domain-containing protein 1 (NOD1) receptor signalling in chicken.

2. Tissue-specific expression analysis of NOD1, receptor-interacting serine-threonine kinase 2 (RIPK2), nuclear factor kappa B (NF-κB) and mitogen-activated protein kinase 11 (MAPK11 or p38) by quantitative real-time PCR (qRT-PCR) revealed their wide distribution in various organs and tissues.

3. Salmonella pullorum infection activated NOD1 receptor signalling in vivo and in vitro, resulting in significant induction of downstream signalling molecules RIPK2, NF-κB/p65, MAPK11/p38 and the effector molecules IL-1b and IL-8.

4. Activation of NOD1 by its agonist bacterial γ-D-glutamyl-meso-diaminopimelic acid (iE-DAP) in HD11 cells induced the adapter molecular RIPK2 and activated the NF-κB/p65 and MAPK11/p38 pathways, resulting in an increase in IL-8 but not IL-1β. Additionally, inhibition of NOD1 using NOD1-shRNA resulted in downregulation of RIPK2, MAPK11 and IL-8, while NF-κB/p65 and IL-1β were unaltered.

5. These results highlight the important role of NOD1 receptors in eliciting the innate immune response following pathogenic invasion in chicken.     

基于DSPACE的藏羌民族文化研究专题库建设

在长期的办学过程中,阿坝师范高等专科学校产生和积淀了有关藏羌民族文化研究的丰富的学术研究成果,通过DSpace系统构建藏羌民族文化研究专题库,可以解决藏羌民族文化成果管理存在展现不足、缺乏长期保存和获取困难的现状,进一步丰富图书馆的馆藏,促进资源的共享,提高学校的学术影响力。     

The evolution of chicken stem cell culture methods

1. The avian embryo is an excellent model for studying embryology and the production of pharmaceutical proteins in transgenic chickens. Furthermore, chicken stem cells have the potential for proliferation and differentiation and emerged as an attractive tool for various cell-based technologies.

2. The objective of these studies is the derivation and culture of these stem cells is the production of transgenic birds for recombinant biomaterials and vaccine manufacture, drug and cytotoxicity testing, as well as to gain insight into basic science, including cell tracking.

3. Despite similarities among the established chicken stem cell lines, fundamental differences have been reported between their culture conditions and applications. Recent conventional protocols used for expansion and culture of chicken stem cells mostly depend on feeder cells, serum-containing media and static culture.

4. Utilising chicken stem cells for generation of cell-based transgenic birds and a variety of vaccines requires large-scale cell production. However, scaling up the conventional adherent chicken stem cells is challenging and labour intensive. Development of a suspension cell culture process for chicken embryonic stem cells (cESCs), chicken primordial germ cells (PGCs) and chicken induced pluripotent stem cells (ciPSCs) will be an important advance for increasing the growth kinetics of these cells.

6. This review describes various approaches and suggestions to achieve optimal cell growth for defined chicken stem cells cultures and use in future manufacturing applications.     

甘加藏羊发情周期下丘脑-垂体-卵巢轴KISS-1/GPR54的表达与分布

为了研究高原甘加藏羊(Ovis aries)发情周期下丘脑-垂体-卵巢轴(hypothalamic-pituitary-ovary axis,HPOA)中肿瘤转移抑制基因(kisspeptins,KISS-1)/G蛋白偶联受体54(G protein-coupled receptors 54,GPR54)的表达及其对藏羊季节性发情周期的调控作用,本实验选取处于发情周期的甘加藏羊32只,应用qRT-PCR和免疫组织化学技术,检测其在发情周期HPOA组织中的表达和分布。qRT-PCR结果显示,甘加藏羊整个发情周期HPOA组织中均有KISS-1和GPR54 mRNA表达。下丘脑组织中发情前期KISS-1 mRNA相对表达量显著(P<0.05)高于其他3个发情时期;在垂体组织中发情前期和间情期的相对表达量较高,与发情后期差异显著(P<0.05);卵巢轴组织中间情期的相对表达量最高,显著(P<0.05)高于其他3个时期。下丘脑组织中发情期GPR54 mRNA相对表达量显著(P<0.05)高于其他3个时期;垂体组织中发情前期的相对表达量显著(P<0.05)高于其他时期;卵巢轴组织中发情后期的相对表达量显著(P<0.05)高于其他时期。免疫组织化学染色结果显示:Kisspeptin和GPR54阳性产物在下丘脑中主要分布于促垂体区的弓状核、脑室前腹侧区;腺垂体中主要在嗜碱性细胞和嫌色细胞中表达;卵巢轴中主要在卵泡颗粒层及卵泡内膜上表达。KISS-1和GPR54在藏羊发情周期HPOA中的差异表达,对藏羊季节性发情周期有一定的调控作用。     

Serum amyloid A (SAA) mRNA expression in chicken and quails in response to bacterial stress

Monitoring of acute phase proteins such as serum amyloid A at gene expression level may provide quick information about immune status of the host and its susceptibility towards common infections. Present study was carried out to evaluate and compare the mRNA expression of SAA gene in Rhode Island Red chicken (RIR) and Japanese quails using real time PCR analysis in response to inactivated Salmonella gallinarum culture. The results showed that expression of SAA gene was approximately 17–33 folds higher in case of birds administered with bacterial culture when compared to un-inoculated controls and expression was higher and quicker in case of quails than RIR chicken. The SAA genes from chicken and quail were cloned and upon sequence analysis it was observed that deduced amino acid sequence of SAA from chicken and quails were having approximately seven percent variation which might have significance in function of this protein in these species.     

基于粪便纤维组织法探究异地驯养藏羚的食性

基于粪便纤维组织法探讨异地(铁卜加草原站)驯养藏羚的食性。结果表明,在基地生长的50种植物中,伊凡苔草、火绒草、川青早熟禾、洽草等是藏羚最喜爱食用植物,所占比例依次为18%、13%、11%和8%,其中伊凡苔草的茎、火绒草的叶、菥冥的果和迷果芹的花是其喜食部位。在基地驯养藏羚的食物组成中,各科所占比例依次为:禾本科(43%)莎草科(25%)菊科(14%)十字花科(7%)豆科(3%)。与可可西里地区类似,铁卜加地区藏羚以单子叶植物为主要食物。