转 Bt基因抗虫棉对棉大卷叶螟抗性的研究

:转Bt基因抗虫棉对棉铃虫有很好的抗性,对棉大卷叶螟(SyleptaderogataFabricius)的抗性未见报道。作者研究表明,棉大卷叶螟在棉田为聚集分布型,低龄幼虫有集中为害习性;抗虫棉对其有很好的抗性,平均虫株率为2.2%,较常规棉中棉所12降低96.6%;百株虫量为133.8头,较常规棉降低88.4%。     

Adult rat and human bone marrow mesenchymal stem cells differentiate into neurons with Musk's polypeptide

AIM: To investigate the differentiation from adult rat and human bone marrow mesenchymal stem cells (BMMSCs) into neuron with musk polypeptide (Mu-P).METHODS: Adult rat and human BMMSCs were induced with Mu-P.Neuron-specific enolase (NSE),neurofilament (NF),Nestin,glial fibrillary acidic protein (GFAP) were detected by immunohistochemistry.RESULTS: Simple methods with Mu-P induced adult rat and human BMMSCs exhibiting a neuronal phenotype,expressing Nestin at 3 hours to 5 hours,and expressing NE and NF at 5 hours to 7 days.But the neuron-like cells didn't express the glial astrocyte marker GFAP.CONCLUSION: Adult rat and human BMMSCs can be induced to differentiate into neurons with Mu-P.     

甲基对硫磷人工抗原的合成与鉴定

根据甲基对硫磷的结构特征,分别从甲基对硫磷的硝基一端或磷酸酯基甲氧基一端引入一定长度的间隔臂和活性基团,设计并合成了4种结构的半抗原;针对各种半抗原所带活性基团的性质,通过多种方法制备了与不同的载体蛋白偶联的人工抗原。进行动物免疫后,得到了针对甲基对硫磷的高效价抗体。     

Regulation of MMP-2 and TIMP-3 expression by uPA signal transduction system in human bone giant cell tumor

AIM: To study effects of urokinase-type plasminogen activator (uPA) signal transduction on expression of matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of matrix metalloproteinase-3 (TIMP-3) in giant cell tumor of bone (GCT). METHODS: Expression of uPAR, MMP-2 and TIMP-3 in GCT tissue was detected by immunohistochemistry. Phosphorylation level of mitogen-activated protein kinase (p44) in uPA/uPAR signal pathway in cultured GCT cells was detected by immunoprecipitation. The expression of MMP-2 and TIMP-3 in cultured cells after treatment with uPA-ATF or anti-uPAR antibody was also detected by Western blotting. RESULTS: 1) Urokinase-type plasminogen activator receptor (uPAR) was positive on the cell membrane and in cytoplasm of some mononuclear stromal cells (MSCs) and multinucleated giant cells (MGCs); 2) MMP-2 was positive in the cytoplasm and on the cell membrane of almost all of MSCs and some of MGCs. The polar distribution of MMP-2 in the cytoplasm of MGCs was especially obvious; 3) The expression of TIMP-3 of some MSCs and MGCs in GCT was much lower than MMP-2. The positive signal also showed a prominent polarity; 4) After treatment with uPA-ATF, the phosphorylation level of p44 in GCT cultured cells was much higher than the control. Addition of anti-uPAR antibody in the cells remarkably down-regulated the phosphorylation level of p44 as compared with the control group, suggesting that uPA-ATF participates cell signal transduction and this reaction can be inhibited by anti-uPAR antibody; 5) uPA-ATF cell signal pathway up-regulated expression of MMP-2 and TIMP-3, while anti-uPAR antibody down-regulated the expression of MMP-2 and TIMP-3. CONCLUSION: These results demonstrate for the first time that uPA-ATF directly regulates the expression of MMP-2 and TIMP-3 by signal transduction pathway, and the over-expression of MMP-2 and TIMP-3 may play an important role in local osteolysis of GCT.     

Relationship of p21WAF1 gene polymorphisms with protein expression in breast carcinomas

AIM: To investigate the relationship between p21^WAF1gene polymorphisms and protein expression in breast carcinoma. METHODS: Polymerase chain reaction single-strand conformation polymorphisms technique (PCR-SSCP) and immunohistochemical assay of S-P immunostaining technique were used to study polymorphisms of p21^WAF1 and protein expression respectively on the specimen of paraffin-embedded tissues in 100 cases of breast carcinomas and 40 benign breast diseases as control. RESULTS: Two p21^WAF1 gene polymorphisms were found in 18% (18/100) of breast carcinomas and 5% (2/40) of control samples. The difference between the two groups was statistically significant (χ²=3.94, P＜0.05). The positive immunohistochemical reaction of p21^WAF1 protein were found in 50% (50/100) of breast carcinomas and 12.5% (5/40) of control samples. The difference between the two groups was statistically significant (χ²=16.84, P＜0.01). The positive immunohistochemical reaction of p21^WAF1 protein were found in 100% (18/18) of breast carcinomas with p21^WAF1 gene polymorphisms and 39% (32/82) of no p21^WAF1 gene polymorphisms. The difference between two groups was statistically significant (χ²=21.95, P＜0.01). The p21^WAF1 gene polymorphisms were correlated with the protein expression in breast carcinomas (r=0.576, P＜0.01). CONCLUSION: p21^WAF1 gene polymorphisms may create the different copies of mRNA and may make relevant protein molecules.     

Induction of tumor-specific cytotoxic T lymphocytes in vitro by dendritic cells pulsed with different glioma antigens

AIM: The goal of this study was to compare different methods for tumor antigen preparation, to observe the induction of tumor-specific cytotoxic T lymphocytes in rats by dendritic cells (DCs) pulsed with different tumor antigens. METHODS: The precursors of dendritic cells were isolated from bone marrow of rats, stimulated in vitro with recombinent rat granulocyte-macrophage colony-stimulating factor (rrGM-CSF) and interleukin-4 (rrIL-4). Then rat DCs were pulsed with C6 tumor cell antigens prepared with different methods: freeze-thaw, boiling or total protein extracted from ultrasonic crushed tumor cell. Subsequently primed DCs were cocultured with T lymphocytes isolated from spleen to induce CTL. Lymphocyte chemoattractant factor from DCs and cytokine IFN-γ release were determined by ELISA, the cytotoxicity of CTL was assayed by JAM test. RESULTS: DCs pulsed with boiled tumor cell in vitro induced an enhanced ability of T-cell proliferation and cytotoxic T lymphocyte activity.CONCLUSION: Our results demonstrated that DCs primed with boiled tumor cell may represent a method for inducing immune responses against the entire repertoire of tumor antigens of malignancies.     

HBV/HAV复合抗原基因在CHO细胞中的表达

为探讨HBV／HAV复合疫苗的可行性，利用DNA重组技术将HBsAg与HAW复合多表位抗原基因VPX进行融合，插入到真核表达质粒pVAX1多克隆位点，构建成核酸表达疫苗pVAX1／SVPX，将其转染CHO细胞。转染细胞培养48h后，进行RTPCR、Western-blot、ELISA分析，结果表明HBV／HAV复合抗原基因SVPX能够在CHO细胞内转录、表达，融合蛋白具有HBV和HAV抗原的特性。     

猪伪狂犬病基因缺失疫苗的制备、安全性、免疫原性、保存期测定及区域试验

为了提供有效的伪狂犬病疫苗，用鸡胚成纤维细胞扩大培养了PrV HB-98突变株（TK^-/gG^-/LacZ^＋），研制了伪狂犬病基因缺失疫苗，并对该疫苗经肌肉接种、经口等免疫途径的最小免疫剂量进行了测定，同时也对4批疫苗的安全性、效力、免疫期和保存期进行了检测；同时将4批疫苗用于免疫23个猪场的母猪、新生仔猪和育肥猪进行区域试验。测定结果表明，疫苗经上述两种途径接种对不同阶段猪的最小免疫剂量均为10^5.0 TCID50；10倍免疫剂量的疫苗对初生仔猪、15日龄仔猪和妊娠母猪是安全的，免疫猪能抵抗强毒的攻击；疫苗在4℃和-20℃下分别可保存6个月和12个月。对伪狂犬病毒抗体阴性的70日龄商品猪和种猪的免疫期为6个月。田间试验表明，4批猪伪狂犬病基因缺失疫苗安全有效，并可用于仔猪发病时的紧急接种。为猪伪狂犬病基因工程疫苗的制备与应用提供了有力的依据。     

民国时期西北地区苜蓿栽培利用刍考

自汉代苜蓿引入我国以来,受到历朝历代的重视,民国时期亦不例外,苜蓿种植利用也得到了发展。在收集近现代有关苜蓿研究文献的基础上,对民国时期西北地区苜蓿种植利用情形进行了梳理与归纳,结果表明,在民国时期西北地区乃为我国苜蓿种植集中区,在陕西、甘肃、新疆、绥远(西部)、宁夏和青海等都有种植,据不完全考查,种植苜蓿的县有52个,其中以陕西最多,达22个县,甘肃次之为14个县,新疆为8个县,绥远(西部)为3个县,宁夏2县(道)和青海1个县。特别是陕甘宁边区发展苜蓿的势头高涨,如1942年边区政府在延安、安塞、甘泉、志丹、定边、靖边等县种植苜蓿达3万亩,陇东种植苜蓿2.3万亩;1944年延川县紫花苜蓿保留面积2.0万亩,到1949年,陕西全省种植苜蓿约98.49万亩。1949年新疆苜蓿保留面积达29300 hm²。绥远河套地区还进行了苜蓿粮草轮作,并建立了苜蓿种植基地。为了鼓励苜蓿种植,边区政府出台了不少政策,例如,1941年边区政府公布了《陕甘宁边区政府建设厅关于种牧草的指示信》,1942年边区政府颁布了《陕甘宁边区卅一年推广苜蓿实施办法》等,李仪祉在治理黄河的方略中,从大农业、生态环境和经济效益出发,提倡广种苜蓿,肥田养畜,并提出了4条种植苜蓿的措施。李烛尘提出,苜蓿根入土深,且能耐旱,适宜西北地区种植和培植草原。苜蓿除用于饲喂家畜外,幼嫩时可当蔬菜食用,在灾荒年也是百姓很好的救荒食物,苜蓿作为农产品常出现在兰州的市场上,试验表明苜蓿保存水土流失的作用要大于作物。     

我国明代苜蓿栽培利用刍考

苜蓿被《救荒本草》《本草纲目》《群芳谱》和《农政全书》以及明皇帝实录与方志等经典要籍所记载,充分体现了苜蓿在明代的重要性、研究的普遍性和种植的广泛性。本研究以记载明代苜蓿的相关典籍为基础,应用植物考据学原理和方法,结合现代研究成果,对明代苜蓿种植分布与状况、植物生态生物学特性、栽培管理和利用方式等进行尝试性研究考查。结果表明,明代在山西、陕西、河北、河南、山东、安徽、江苏、北京、甘肃和宁夏等省均有苜蓿种植,其中以“三晋为盛,秦、鲁次之,燕、赵又次之”。在苜蓿植物学、生态生物学研究方面成绩显著,对苜蓿植株形态、花色及其着生部位、荚果种子形状的精准描述已达到现代植物学的水准。同时,对苜蓿的轴根性也有一定的认识,记载其根的形态与黄芪的根相类似。在明代出现了紫花和黄花2种苜蓿的记载;主张苜蓿与荞麦混作,并利用苜蓿的肥田能力,将苜蓿纳入了轮作制度中;提倡7、8月种苜蓿,一年三刈,种子田一刈;苜蓿3年后生长进入旺盛期,7、8年后衰退垦去。在苜蓿饲用方面明代王象晋提出了最佳利用时期,即“苜蓿花时,刈取喂马牛,易肥健食”。同时,在苜蓿的食用、药用等方面人们利用得更加具体有效。此外,苜蓿还可做贡品。