多重PCR检测猪细小病毒和猪伪狂犬病病毒的研究

根据GenBank上已发表的猪细小病毒（Porcine parvovirus，PPV）的VP2基因序列和猪伪狂犬病病毒（Porcine pseudorabies virus，PRV）的gH基因序列，设计合成了两对特异引物，分别建立了PPV和PRV的单项PCR诊断方法，通过对扩增条件的筛选，最终成功地建立了PPV和PRV的复合PCR诊断方法，即利用一次PCR反应，可同时扩增PPV的751bp和PRV355bp的特异性片段，而扩增猪圆环病毒Ⅱ型（PCV．2）及相应的培养细胞（PK-15）核酸结果均为阴性，对PPV和PRV的最低检出量分别为100Pg和10Pg的DNA。该方法适合对PPV和PRV的联合检测和鉴别诊断。     

犬瘟热病毒自然感染犬淋巴组织中T、B细胞变化的研究

为了调查患犬瘟热病犬淋巴组织中T、B细胞变化的特点及淋巴细胞减少的发病机制,试验通过免疫组织化学的方法观察了T细胞(用CD3和CD45RO检测T细胞)、B细胞(用IgG、IgM抗血清检测B细胞)和犬瘟热病毒(抗犬瘟热病毒抗体)在病犬淋巴组织中的分布。结果表明:在淋巴组织中的淋巴细胞、淋巴小结中树突状细胞和巨噬细胞中均检出了抗病毒阳性反应细胞。在骨髓组织的前髓细胞中也发现抗病毒阳性反应细胞和嗜酸性胞浆内及核内包涵体的存在。与对照组相比,CD3和CD45RO阳性细胞主要存在于T细胞的分布域;但CD3和CD45RO阳性T细胞的数量较少。位于淋巴组织中的巨噬细胞有的被CD45RO染成阳性。在B细胞分布的区域中,IgG、IgM阳性细胞的数量明显减少;一些位于淋巴组织的浆细胞也被IgG或IgM染成阳性。在淋巴组织中淋巴细胞减少的顺序为:IgG阳性细胞减少最明显,其次为IgM和CD45RO阳性细胞,再次为CD3阳性细胞。依据试验结果,作者认为病犬淋巴组织中淋巴细胞减少主要是由B细胞缺乏所引起的;淋巴细胞的增殖能力减弱是引起淋巴组织中淋巴细胞减少的重要原因。     

增强猪传染性胸膜肺炎灭活苗对小鼠保护力的研究

为了提高猪传染性胸膜肺炎灭活苗的保护力,本研究利用基因重组技术表达了猪传染性胸膜肺炎放线杆菌6种主要致病因子蛋白:rApxⅠ、rApxⅡ、rApxⅢ、rApxⅣ、rApfa和rOMP。设立试验Ⅰ组(灭活苗),试验Ⅱ组(灭活苗、rApxⅠ、rApxⅡ、rApxⅢ和rOMP),试验Ⅲ组(灭活苗、rApxⅠ、rApxⅡ、rApxⅢ和rApxⅣ),试验Ⅳ组(灭活苗、rApxⅠ、rApxⅡ、rApxⅢ和rApfa)和空白对照组(PBS),每组26只BALB/c小鼠,分3次免疫,每次间隔2周,每周检测血清抗体水平,在免疫后第2周、第4周和第5周,MTT法检测脾脏T细胞增殖水平及ELISA法测定IFN-γ分泌水平。在第3次免疫后的第7 d,将每组剩余20只小鼠平均分成2组,分别以APP1型菌(5×10~9 CFU)和APP7型菌(1×10~(11) CFU)进行攻毒保护试验。结果表明,向灭活苗中添加重组亚单位成份可以提高机体的细胞免疫和体液免疫水平,并能提高灭活苗的交叉保护率;试验Ⅱ组的ApxⅠ和OMP抗体水平以及细胞免疫水平均显著高于其它各组(p<0.05),对于APP1和APP7型菌的攻毒也提供了明显优于其它各组的保护结果,试验Ⅲ、Ⅳ组的细胞、体液免疫水平及保护率较试验Ⅰ组也有所提高。结果显示出ApxⅠ、OMP抗体水平和细胞免疫水平与攻毒保护率之间存在着正相关性,向灭活苗中加入重组蛋白成份可以提高疫苗的交叉保护,试验Ⅱ组通过激活体液免疫和细胞免疫,对两个血清型APP的攻击提供了很好的保护作用,从而为猪传染性胸膜肺炎新型疫苗的研制提供参考。     

猪囊尾蚴循环抗原和排泄分泌抗原研究进展

猪囊尾蚴病是由带科(Taeniidae)带属(Taenia)的猪带绦虫(有钩绦虫)(Taenia solium)的幼虫-猪囊尾蚴(Cysticercus cellulosae)寄生于猪的肌肉和其他器官中而引起的一种人畜共患寄生虫病,俗称囊虫病。该病主     

梨果实贮藏中可溶性果胶和半纤维素分子结构的变化

 为了了解分子结构变化与耐藏性的关系，以耐贮性不同的苹果梨和新高梨为试材，利用Ultrogel AcA 22凝胶柱渗漏分离，比较了梨贮藏中可溶性果胶和半纤维素分子大小的变化。结果表明，苹果梨的半纤维素分子大于新高梨。在贮藏过程中，耐贮性强的苹果梨果实可溶性果胶分子大小变化不明显，但半纤维素分子明显变小。相反，新高梨的可溶性果胶分子明显变小，而未观察到半纤维素分子大小的变化。说明梨的耐贮性与可溶性果胶分子结构变化及半纤维素分子大小有关，而与半纤维素分子结构变化没有直接关系。     

Classification of Morchella species from Yunnan and Zhejiang Provinces Based on rDNA ITS Sequences

Fourteen Morchella samples (eleven from Yunnan and three from Zhejiang Provinces) were selected on the basis of differences in fruit body morphology. Ribosomal DNA internal transcribed spacers (ITS) were amplified in each case using the universal primer pair, ITS-1 and ITS-4, and the amplification products were purified and sequenced. Comparisons with sequence data in GenBank revealed that the 11 Morchella isolates collected from Yunnan belonged to four species: Morchella elata, Morchella conica, Morchella crassipes and Morchella costata. The three isolates collected from Zhejiang Province (M12, M13 and M14) were designated as unknown Morchella species. When Verpa conica (AJ544206) (from the genus Verpa belonging to the same family as Morchella) was taken as the outgroup, the 14 isolates formed three groups, M. elata, M. costata (Group 1); Morchella esculenta, M. conica (Group 2); and M. crassipes, M12, M13 and M14 (Group 3).     

切花菊14个品种的核形态学研究

 采用常规压片法对不同花序类型、不同花色的14个切花菊品种进行了核形态学研究。结果表明：（1）间期核均为复杂染色中心型，前期染色体属于中间型。(2)体细胞染色体在品种内和品种间均呈现多数目性，染色体数变动范围在48~56之间，以54条为主。（3）核型呈现多样性，由中部、近中部和近端部着丝粒染色体组成，‘神马’、‘小红娘’、‘九月黄’和‘小菊双色’有1或2条随体染色体；染色体相对长度为2.04%~5.26%；核型不对称系数为62.39%~67.32%；‘长紫’、‘意大利红’、‘小红娘’和‘阿里格斯’核型类型为“2B”，其它品种为“2A”型。（4）有丝分裂后期和末期除‘清露’外均正常。基于观察结果分析了切花菊核型多样性及进化趋势。     

Biological characteristics of JAK2 transduced CD34+ cells from cord blood during ex vivo expansion

AIM: To explore the feasibility and biological characterization of long-term regulated expansion of JAK2 transduced human CD34⁺ cord blood cells in vitro.METHODS: A retrovirus (RV) vector which contains JAK2 catalytic domain and two binding sites for a chemical inducer,dimerization (AP20187),was cloned (designated MGI-F₂JAK2).CD34⁺cells were enriched from cord blood with a MiniMACS system.The purified CD34⁺ cells were transfected with supernatant from the retrovirus packaging cell line that expressed JAK2.Following transduction,cells were expanded into four groups: AP20187 alone,FL alone,TPO,alone,AP20187+FL+TPO,respectively.The expanded cells were monitored by GFP expression,immunophenotyping,progenitor colony assay,karyotype analysis as well as tumorigenesis in nude mice.RESULTS: The purity of selected CD34⁺ cells was over 91% and gene transfer rate was 49.32%±6.21%.Only the group of AP20187 +FL+ TPO was obtained a significant sustained outgrowth of the transduced CD34⁺ cord blood cells.The percentage of GFP+ cells consistently produced a rise to the 90% peak level by the end of 8th week of culture.Flow cytometry analysis showed that the phenotype of the expanded cells was CD33⁺,CD61⁺ and Gly-A⁺ partial positive;CD38⁺ and HLA-DR⁺ strong positive,while CD2,CD7 and CD19 were almost negative.Colony assays performed in methycelluos,which can give rise to BFU-E,CFU-GM and CFU-Mix,the CFU-GM was predominantly in all colonies.The tumor was not observed in nude mice and the karyotype analysis was normal from expanded cells.CONCLUSION: The results demonstrate that AP20187-mediated activation of JAK2 signaling is capable of stimulating expansion JAK2 transduced CB CD34⁺ cells in combination with FL and TPO.This system may have applications for studies in signaling transduction,hematopoiesis,and for gene and cell therapy.     

Effects of cinnamyl aldehyde on c-Fos and c-Myc expression in NIH3T3 cells

AIM: Cinnamyl aldehyde (CA) is one alcohol ingredient derived from Cinnamomum cassia,which is widely used in treating chronic skin wound in Chinese medicine with the curative effect of ‘rescuing YANG’.The purpose of the present study was to investigate the expression of c-Fos,c-Myc proteins at different time points in NIH3T3 treated with CA and explore the possible mechanism of promoting cell proliferation by CA.METHODS: MTT assay was used for observing cell proliferation.Expression of c-Fos and c-Myc proteins in NIH3T3 cells were assessed by immunocytochemistry assay.RESULTS: The cell proliferation was promoted obviously when CA concentration was between 8.8×10^-2 μg/L and 8.8×10 μg/L.CA at concentration of 5.5 μg/L significantly induced expression of c-Fos,c-Myc proteins at 2-3 h after the stimulation compared with control group (P<0.01).CONCLUSION: CA increases expression of c-Fos and c-Myc proteins,which may be one of mechanisms for CA to promote NIH3T3 cell proliferation.