C3d3通过促进脾脏B细胞高表达CXCR4增强hCGβ DNA免疫体液免疫效应

本文旨在探讨分子佐剂C3d3与hCGβ融合在基因免疫中增强抗hCGβ体液免疫效应的机制。分别用质粒pCMV4-hCGB-C3d3、pCMV4-hCGβ和pCMV4免疫BALB/c小鼠,间接ELISA法检测免疫小鼠外周血IgG/IgA类抗hCGβ抗体水平;ELISPOT分析免疫鼠脾脏组织IgG/IgA类抗体分泌细胞水平(ASC);RT-PCR分析免疫鼠脾脏B细胞趋化因子受体表达,RT-PCR和FCM分析CXCR4表达水平;RT-PCR和ELISA检测脾脏组织CXCL12表达水平。结果显示,pCMV4- hCGβ-C3d3免疫组外周血IgG类抗hCGβ抗体水平明显高于pCMV4-hCGβ免疫组;而IgA类抗hCGβ抗体水平在两组间无明显差异。pCMV4-hCGβ-C3d3免疫组脾脏组织IgG类ASCs水平明显高于pCMV4-hCGβ组;两组间IgA类ASCs水平无明显差异。经pCMV4-hCGB、pCMV4-hCGβ- C3d3免疫鼠脾脏B细胞CXCR4表达明显高于对照组;且pCMV4-hCGβ-C3d3组明显高于pCMV4-hCGβ免疫组。CXCR4~ 细胞与ASCs呈正相关,r=0.966,(P<0.05)。pCMV4-hCGβ-C3d3和pCMV4-hCGβ组脾脏组织CXC L12表达均显著高于对照组。结果表明,分子佐剂C3d3与hCGβ基因融合,在基因免疫小鼠后能够显著升调节脾脏ASCs CXCR4表达,从而可能增强抗hcGβ基因疫苗的体液免疫效应。     

喉癌组织中血管内皮生长因子、CD1a阳性树突状细胞表达及相关性的研究

目的:观察喉癌病人癌组织内CDla~ 树突状细胞(dendritic cells,DC)的分布、形态学特征以及血管内皮生长因子(vascular endothelial growth factor,VEGF)的表达情况,同时探讨喉癌组织CDla~ DC分布与VEGF表达的关系。方法:采用抗VEGF、抗CDla抗体进行免疫组化染色和透射电镜等方法研究了42例喉癌组织。结果:喉癌组织CDla~ 树突状细胞树突状细胞形态不规则,表面有许多不规则树状突起。大部分散在分布于癌巢内,与肿瘤细胞有密切接触,少量分布于癌巢之间的间质和癌周组织。喉癌组织内CDla~ DC密度与喉癌临床期次呈明显的负相关,而VEGF的表达与喉癌临床期次呈明显的正相关。喉癌组织中VEGF表达明显升高的病例,其CDla~ DC密度显著降低。结论:癌巢内树突状细胞为不成熟状态的DC,与肿瘤细胞密切接触而捕获肿瘤抗原。喉癌组织中VEGF表达与DC含量呈负相关。     

尘螨变应原Derf1核酸序列测定及其分子进化分析

目的:对尘螨主要变应原Der f1进行核酸序列测定,探讨其系统进化信息。方法:根据Genbank公布的Der f1基因序列设计引物,巢式PCR扩增Der f1的cDNA,纯化、回收、克隆至pMD19-T simple后进行序列测定,序列比对后用Clustal W 1.83构建分子进化树。结果:成功扩增出Der f1的cDNA片段,测序表明该基因含ORF1个,长度966bp,与参考序列同源性达99.9%。该变应原具半胱氨酸蛋白酶活性,与果蝇进化关系最远,与梅氏嗜霉螨进化关系最近。结论:成功获得了尘螨变应原Der f1基因片段,根据其编码的氨基酸序列构建出的系统进化树与形态学分类不一致。     

金皮猪肉质相关组织学特征与CAST基因HinfI多态性的关系研究

本实验选择具有优良肉质的本地金华猪与肉质较差的引进品种皮特兰猪杂交所产的金皮F2代为研究对象,利用PCR-RFLP的方法检测金皮F2代猪CAST基因型的频率,并以肌球蛋白重链（MHC）免疫组织化学、过碘酸希夫反应和苏木精-伊红等多种组织学方法,研究猪背腰最长肌的肌肉组织学特性,并对其进行图像分析,在此基础上采用SAS分析其相互关系。结果发现．PAS染色后肌纤维可区分为三种类型。PAS阴性（Ⅰ）、PAS反应强阳性（Ⅱ-a）和PAS反应强度中间型（Ⅱ-b）。利用肌球蛋白重链单克隆抗体MHCs和MHCf染色结果表明．金皮F2代杂交猪背腰最长肌MHCs阳性（慢肌）纤维的比例为7．62％,快肌纤维为92.38％。CAST基因的扩增产物具有524bp和632bp两个Hinfl多态性片段,定义等位基因为A（114＋192＋400＋632bp）,B（114＋192＋400＋524bp）。AA、BB和AB基因型频率分别为0．1795、0．5897和0．2308,A和B的基因型频率分别为0．4743和0．5256。AA、BB和AB单型对眼肌面积．系水率、pH值、导电率等参数均无显著的影响。CAST基因的HinfI多态性对肌纤维的轴比有显著影响,AA单型有产生较多轴比较大（近似椭圆形）肌纤维的倾向．而BB则控制形成更多的轴比更小（近似于圆形）的肌纤维。具有AA单型的个体具有更多的肌间结缔组织．而具有BB单型的个体的肌间结缔组织较少．AB单型的个体介于两者之间。     

控制水稻叶片叶绿素含量及其离体叶片叶绿素降解速度相关的QTL定位(简报)

许多研究认为,在一定范围内,叶绿素含量与光合速率成正相关关系、叶绿素含量高的水稻叶片能延缓衰老。理论上推算,水稻叶片如果推迟1天衰老,可使水稻增产2％左右,而实际实验结果表明可增产1％左右。叶片早衰往往也是造成有些水稻品种结实率偏低、空秕率较高及产量降低的主要原因。叶片衰老是水稻发育过程中的生命现象,它是水稻在长期进化过程中形成的适应性。叶片衰老的显著特征之一是叶绿素含量下降,叶色褪绿变黄。[第一段]     

芨芨草愈伤组织器官发生及植株再生

芨芨草(Achnatherum splendens (Trin.) Nevski)种子消毒并在MS培养基上萌发获得无菌苗, 以幼苗的叶鞘和胚轴为外植体诱导愈伤组织, 经继代后进一步诱导不定芽及生根。研究结果表明, 诱导愈伤组织最适合的培养基为B5+1.5 mg.L-12,4-D+0.5 mg.L-1 NAA; 诱导芽分化较适合的培养基为B5+0.5 mg.L-1 6-BA +0.2 mg.L-1 NAA; 1/4 B5+1.0 mg.L-1 NAA+0.2 mg.L-1 IBA +1.0 g.L-1活性炭培养基则有利于芨芨草试管苗的生根。本实验建立了完整的芨芨草植株再生体系, 移栽成活率高。     

Biological effects of low-energy C ion implantation on sesame (Sesamum indicum L.)

Field cultivation experiments on white sesame (Sesamum indicum L.)seeds implanted with low-energy C ion showed that different dosages of C ion implantation produce different biological effects.Sesame plants in 6 different dosage groups with C ion density respectively at 1×1011,1×1012,1×1015,5×1015,1×1016,5×1016 ion/cm2 were superior to the control group in plant height,leaf number,stalk diameter and leaf size.Further,sesame plants in these groups flower and seed earlier than those in the control group,and single plant yield also increased.Of all the groups,the 5×1015 ion/cm2 dosage group yielded the best effect,whereas the 1×1017/cm2 dosage group showed an evident inhibitory effect of ion implantation on the germination and growth of the sesame seeds.     

Quantitative determination of trans-polydatin, a natural strong anti-oxidative compound, in rat plasma and cellular environment of a human colon adenocarcinoma cell line for pharmacokinetic studies

A simple, accurate, precise, specific and reproducible high-performance liquid chromatography (HPLC) method was developed for determination of trans-polydatin, a natural strong anti-oxidative compound, in rat plasma and cell suspension. The assay procedure involved simple liquid-liquid extraction, the supernatant liquid was added an equal volume of water to avoid solvent effect. The detection of the analyte peak was achieved by monitoring the eluate using a UV detector set at 303 nm. The analysis used a Hypersil ODS2 C18 column (5 microm, 4.6 mm x 250 mm) and methanol/distilled water as the mobile phase (flow rate=1 mL/min). A total analytical run was achieved within 6.0 min and calibration curve was linear over a wide concentration range of 0.25-40 microg/mL for plasma sample and 1.0-500 microM for cell suspension, the coefficients of correlation were 0.9997 and 0.9999 or better, respectively. There was 80.7+/-7.86%, 96.8+/-3.20% and 102.7+/-9.72% recovery from 0.5, 10, and 40 microg/mL plasma samples, respectively. Intra- and inter-batch accuracy and precision were acceptable for the both matrices. The RSD of intra- and inter-day assay variations were all less than 10%. Both analyte and IS were stable in the battery of stability studies, freeze-thaw cycles. The described assay method was applied to pharmacokinetic studies in rats and a human colon adenocarcinoma cell line (Caco-2) successfully. The application of the assay to determine the pharmacokinetic is described.     

Antisense RNA-mediated suppression of Bmi-1 gene expression inhibits the proliferation of lung cancer cell line A549

The oncogene Bmi-1 regulates cell proliferation and senescence. It is reported that it controlled the self-renewal of leukemic and breast cancer stem cell and was overexpressed in some solid tumors and hematologic malignancies. In this study, the effects of inactivation of Bmi-1 mediated by a plasmid-expressing antisense Bmi-1 RNA on the proliferation of lung cancer cell line A549 were investigated. As a result, when the plasmid was stably introduced into the cell line, the Bmi-1 protein level was specifically downregulated, and the cell proliferation was significantly inhibited as shown by the cell growth curve and colony forming assay. The cells were found mostly in the phase of G(0)/G(1) and cells in S phase were significantly decreased. Our results suggest that targeting Bmi-1 might be a therapeutic potential for the treatment of non-small-cell lung cancer.