表达性

The expression level of tim gene in 25LL is markedless higher than 15DD and 20LD in the early incubation stage(7 days before), but it is obviously higher than 20LD and 15DD conditions in the late.To further invesgate the per and tim expression in different moment every day of the sensitive period(7d,8d,9d) during the process of embryogeny,the expression of tim gene was very low and the per and tim expression did not have obvious rhythmicity under 15DD condition.

进一步调查家蚕胚胎对环境敏感时期(7d-9d)per和tim基因的昼夜表达节律，在15DD条件下，tim基因表达量一直很低，tim基因和per基因的表达量都没有明显的节律性变化；而25LL和20LD条件下，两基因在一日中不同时刻的表达具有明显的节律性，表达量也较高。

The recombinant proteins form inclusionbody in cytoplasm. Dissolve the inclusionbody with urea and refold it with metal affinity chromatography. The refold recombinant protein has certain antibacterial activity.

对表达产物进行复性，结果显示无论是融合表达的VHb-Fetidin还是非融合表达的Fetidin都无法用稀释法和尿素浓度梯度透析法成功复性，只有VHb-Fetidin可以通过金属亲和层析柱成功复性。

After analyzing the merit and lack of 9I model, the main idea is proposed: to distill some basic topological relations from 9I model, of which the composite topological relations of complex objects are made up. In detail, the composite topological relationships between region and region, line and line, line and region are particularly discussed;(2)Analyze the characteristics and inner relations of cadastral features. According to the expressional model of complex topological relations, the author discovers the possible topological relations between parcels, parcel lines and parcel points. In addition, sum up the general topological relationship rules of cadastral data.(3) Then the topological relations model is extended to the spatio-temporal data. Time in cadastral database and change semantic are discussed. It's redefined that spatio-temporal topological relations is composed of time, location, status and event relation. Spatio-temporal topological relationships can be represented by extended 4I model. Taking cadastral parcels as example, there are different possible spatio-temporal topological relationships between parcels when they are extended, shrunken, split, merged or others. Thus, some deductions are drawn that, for example, if parcels have be coexisting for some time they are impossible to be overlapped. Furthermore, the parents of parcels alternated and their possible change types can be detected by their spatio-temporal topological relationships.(4) Before carrying out the experiment, the calculation method must be designed.

具体研究工作及成果如下：（1）根据点集拓扑理论中与拓扑空间相关的概念，本文对空间的点、线、面进行了重新定义，以区分对象的复杂性；分析了9I模型在表示简单对象方面的优点，以及在表达复杂对象上的不足，阐述了本文的研究思路和解决方案，并以复杂的面面、线线和线面为例详细探讨了复合拓扑关系的表达方法；（2）分析了地籍权属数据和地类数据的内涵、特点和要素之间的联系，根据复杂对象的拓扑关系表达模型，本文讨论了宗地、界址线、界址点之间，图斑、线状地物、零星地类之间可能存在的拓扑关系和形式化表达方法，归纳总结了地籍对象的拓扑关系满足的约束性规则；（3）将拓扑关系的研究延伸到时空领域，从地籍对象的时态性出发，本文阐述了地籍数据中时间的含义和变化语义描述的需要；从时间、位置、状态、事件等之间的拓扑关系方面，重新系统性地进行了时空拓扑关系的定义；研究了一维时间和二维空间的集成表达方式，以宗地、界址线为例，分析了变更前后不同时间区间里可能的拓扑关系，探讨了时空拓扑关系对时空变化操作类型和变化语义的推理方法和意义；（4）为实现空间拓扑关系的查询分析等应用，本文研究了空间拓扑关系的计算方法。

The stable clones are further identified by RT-PCR and Western blot; 6 MTT assay is used to investigate the effect of ZNRD1 on the cell growth of cells (AGS, SGC7901, MKN28, NIH3T3, GES-1); 7 Soft agar assay is used to investigate the effect of ZNRD1 on the clonality of cells (AGS, MKN28); 8 Nude mice assay is used to investigate the effect of ZNRD1 on the cell growth of gastric cancer cells (AGS, MKN28); 9 Flow cytometry is used to investigate the effect of ZNRD1 on the cell cycle distribution of cells (AGS, MKN28, NIH3T3, GES-1); 10 Flow cytometry is used to investigate the effect of ZNRD1 on the cell apoptosis of cells (AGS, MKN28, NIH3T3); 11 MTT assay is used to investigate the effect of ZNRD1 on the drug sensitivity of cancer cells (SGC7901, SGC7901/VCR, HL-60, HL-60/VCR) in vitro; 12 SRCA is used to investigate the effect of ZNRD1 on the drug sensitivity of gastric cancer cells (SGC7901, SGC7901/VCR) in vivo; 13 Flow cytometry is used to investigate the effect of ZNRD1 on adriamycin accumulation of cancer cells (SGC7901, SGC7901/VCR, HL-60, HL-60/VCR); 14 Transmission electron microscope is used to investigate the effect of ZNRD1 on the sensitivity of SGC7901 cells towards drug-induced apoptosis; 15 Flow cytometry and DNA ladder assay are used to investigate the effect of ZNRD1 on the sensitivity of cells (SGC7901, SGC7901/VCR, HL-60/VCR) towards drug-induced apoptosis; 16 Microarray is used to investigate the profiling of ZNRD1-responsive genes in gastric cancer cells (AGS, MKN28, SGC7901, SGC7901/VCR); 17 RT-PCR and Western blot are used to identify the results of microarray; 18 Reporter gene assay is used to investigate the effect of ZNRD1 on the transcriptional activity of cyclin D1; 19 Reporter gene assay is used to investigate the effect of ZNRD1 on the transcriptional activity of MDR1; 20 Kinase assay is used to investigate the effect of ZNRD1 on the activity of cyclin E-CDK2 kinase; 21 The antisensenucleic acids of p21 is used to inhibit the expression of p21, and flow cytometry is used to investigate the effect of p21 on ZNRD1-induced cell cycle arrest in gastric cancer cells; 22 The antisensenucleic acids of p27 is used to inhibit the expression of p27, and flow cytometry is used to investigate the effect of p27 on ZNRD1-induced cell cycle arrest in gastric cancer cells; 23 Liposome is used to up-regulate the expression of Skp2, and flow cytometry is used to investigate the effect of Skp2 on ZNRD1-induced cell cycle arrest in gastric cancer cells; 24 Western blot is used to investigate the effect of ZNRD1 on the stability of Skp2 and p27 in gastric cancer cells; 25 MVD assay is used to investigate the effect of ZNRD1 on the angiopoietic activity of gastric cancer cells; 26 ELISA is used to investigate the effect of ZNRD1 on the expression of VEGF165 in gastric cancer cells; 27 The roles of DARPP-32 in MDR of gastric cancer cells are investigated using gene transfection, MTT assay, SRCA, flow cytometry and DNA ladder assay.

应用杂交瘤技术制备ZNRD1的首个单克隆抗体；2）利用RT-PCR、Western blot和免疫组化检测ZNRD1在胃癌组织、胃炎组织、正常胃上皮组织、胃癌细胞和正常胃组织上皮细胞中的表达；3）构建ZNRD1的小干扰RNA载体，并测序鉴定；4）利用脂质体将ZNRD1的真核表达载体及其空载体转染胃癌细胞（AGS、SGC7901、MKN28）和小鼠成纤维细胞（NIH3T3），G418筛选后进行鉴定；5）利用脂质体将ZNRD1的小干扰RNA载体及其空载体转染药敏胃癌细胞（SGC7901）、正常胃组织上皮细胞（GES-1）、对长春新碱耐药的胃癌细胞（SGC7901/VCR）、药敏白血病细胞（HL-60）、对长春新碱耐药的白血病细胞（HL-60/VCR），G418筛选后进行鉴定；6）利用MTT实验检测ZNRD1高/低表达对细胞（AGS、SGC7901、MKN28、NIH3T3、GES-1）生长的影响；7）通过软琼脂克隆形成实验检测上调ZNRD1对AGS、MKN28细胞克隆形成能力的影响；8）通过裸鼠成瘤实验检测上调ZNRD1对AGS、MKN28细胞体内成瘤性的影响；9）通过流式细胞仪分析ZNRD1高/低表达对细胞（AGS、MKN28、NIH3T3、GES-1）的细胞周期的影响；10）通过流式细胞仪分析上调ZNRD1对细胞（AGS、MKN28、NIH3T3）的凋亡的影响；11）通过MTT实验检测ZNRD1高/低表达对细胞（SGC7901、SGC7901/VCR、HL-60、HL-60/VCR）体外药物敏感性的影响；12）通过肾包膜下移植法检测ZNRD1高/低表达对细胞（SGC7901、SGC7901/VCR）体内药物敏感性的影响；13）通过流式细胞仪分析ZNRD1高/低表达对细胞（SGC7901、SGC7901/VCR、HL-60、HL-60/VCR）内阿霉素蓄积和泵出的影响；14）通过透射电镜检测上调ZNRD1对SGC7901细胞凋亡敏感性的影响；15）通过流式细胞仪和DNA梯度试验检测ZNRD1高/低表达对细胞（SGC7901、SGC7901/VCR、HL-60）凋亡敏感性的影响；16）通过基因芯片检测ZNRD1高/低表达对胃癌细胞内基因表达谱的影响；17）利用RT-PCR、Western blot对基因芯片的结果进行鉴定；18）利用报告基因实验检测ZNRD1对cyclin D1的启动子活性的调节作用；19）利用报告基因实验检测ZNRD1高/低表达对MDR1的启动子活性的调节作用；20）利用激酶试验检测ZNRD1对cyclin E-CDK2 激酶活力的影响；21）利用反义核酸技术抑制p21的表达；通过流式细胞仪检测抑制p21对ZNRD1介导的细胞周期阻滞的影响；22）利用反义核酸技术抑制p27的表达；通过流式细胞仪检测抑制p27对ZNRD1介导的细胞周期阻滞的影响；23）利用脂质体转染法上调Skp2的表达；通过流式细胞仪检测上调Skp2对ZNRD1介导的细胞周期阻滞的影响；24）利用Western blot检测ZNRD1对p27和Skp2的蛋白稳定性的影响；25）利用微血管密度实验检测ZNRD1对AGS、MKN28细胞裸鼠移植瘤微血管形成的影响；26）利用ELISA检测ZNRD1对AGS、MKN28细胞培养上清和移植瘤匀浆中VEGF165含量的影响；27）利用脂质体转染法、MTT实验、肾包膜下移植法、流式细胞仪和DNA梯度试验检测新耐药相关分子DARPP-32对细胞（SGC7901、SGC7901/VCR、对阿霉素耐药的胃癌细胞SGC7901/ADR）多药耐药表型的影响；利用脂质体转染法和MTT实验检测下调ZNRD1对DARPP-32介导的胃癌多药耐药的调控作用。

Results showed:(1) The airway responsiveness of smoke control group was higher than that of normal control group, the airway responsiveness of smoke + exercise group was lower than that of smoke control group;(2) Plasma level of cortisol determined immediately after exercise was higher than that determined before exercise, no significant difference occurs between that determined next day morning after all exercise finished in contrast to that determined before exercise;(3) HE staining showed, there was severe chronic pulmonary inflammatory response in smoke control group, which was reduced in the smoke + exercise group;(4) chronic cigarette smoke downregulated the protein and mRNA expression of BKca in smooth muscles of bronchi and bronchioli, exercise increased the mRNA expression of BKca;(5) chronic cigarette smoke downregulated the protein and mRNA expression of Kv1. 5 in smooth muscles of bronchi and bronchioli, exercise increased both the protein and mRNA expression of Kv1. 5 in contrast to smoke control group in bronchioli and did not effect the expression of Kv1. 5 in bronchi;(6) chronic cigarette smoke induced upregulation of FIZZ1/RELMα in bronchial smooth muscle cells, exercise decreased these effect.

结果：(1)吸烟对照组的气道反应性明显增高，吸烟加运动应激可明显降低大鼠的气道反应性；(2)吸烟运动组运动后血浆皮质醇浓度明显高于运动前，但全部运动结束后次日晨测定的血浆皮质醇浓度与运动前无明显差异；(3)HE染色显示，吸烟对照组肺组织出现明显的慢性炎症反应，吸烟加运动应激组比吸烟对照组炎症反应轻；(4)慢性吸烟可降低大鼠大气道和小气道BKca mRNA和蛋白表达，而吸烟加运动应激可使BKca mRNA表达上调，但对小气道的蛋白表达无影响；(5)慢性吸烟可降低大鼠大气道和小气道Kv1.5蛋白和mRNA表达，在小气道吸烟加运动应激可减轻这种作用，大气道则无作用；(6)吸烟使支气管平滑肌的FIZZ1/RELMα蛋白表达明显增强，而吸烟加运动组的RELMα蛋白及mRNA均明显减弱。

Objective To study the relationship of endangium proliferation and the expression of metalloproteinases and tissue inhibitors of metalloproteinases after angioplasty of iliac artery in rabbits to investigate the probable mechanism of composite Danshen pill in prevention and treatment of restenosisMethods 30 white male rabbits of Japan （the average body weight was 25～30 kg）were randomly divided into 3 groups:normal control group,model group (intima destroyed by balloon and given hypercholesterol diet),CDP group (intima destroyed by balloon and given hypercholesterol diet plus drug CDP 150 mg/d),10 ones each groupResults The results of iliac artery angiogram and the analysis of pathology:there were lumens stenosis,thinner intima,smaller intima area,in CDP group compared with those in model group,and ratio of intima and tunica media thickness and area among each group (P＜001)Immunohistochemistry analysis:the MMP2,MMP9,TIMP1,TIMP2 in model group significantly increased than those in control group (P＜001)The CDP group had lower MMP2 and MMP9 expression,higher TIMP1 and TIMP2 expression,and increased MMP2/TIMP1,MMP9/TIMP2 than model group (P＜001)The thickening of vascular neointima and stenosis degree of vascular lumen were relevant to MMP2 and MMP9 (r=0896,P＜001)Conclusions MMP and TIMP play the very important role in the restenosis process of arteryCDP could inhibit the thickness of vascular neointima after balloon injury,the probable mechanism of which may be inhibiting collagen formation,smooth muscle immigration and decreasing hyperplasia of intima by interfering expression of MMPs and TIMPs

目的 研究兔髂动脉成形术后血管内膜增生和基质金属蛋白酶及其抑制物表达之间的关系，探讨复方丹参滴丸预防再狭窄的可能机制。方法健康成年二级雄性日本大耳白兔30只，平均体重25～30 kg。随机分为3组：正常对照组10只，模型组10只（球囊内膜剥脱加高胆固醇饮食），治疗组10只（球囊内膜剥脱加高胆固醇饮食以及复方丹参滴丸150 mg/d）。结果兔髂动脉造影、血管病理图像分析检测结果：①复方丹参滴丸组较模型组血管造影示管腔直径狭窄、内膜厚度减少、内膜面积减少、内膜厚度和中膜厚度比、面积比，各组之间有显著性差异(P＜001)。②免疫组化分析：模型组MMP2、MMP9、TIMP1、TIMP2表达均高于对照组（P＜001）；复方丹参滴丸组MMP2、MMP9的表达低于模型组，而TIMP1、TIMP2的表达高于模型组；TIMP1/MMP2、TIMP2/MMP9明显增加，差异有显著性（P＜001）。③内膜的增生以及管腔的狭窄程度与MMP2、MMP9有很好的相关性(r=0896，P＜001)。结论 MMP和TIMP在再狭窄形成中起重要作用；复方丹参滴丸能够明显抑制血管损伤后的内膜增生，可能的机制是通过影响金属蛋白酶MMP2、MMP9及其抑制物TIMP1、TIMP2的表达从而抑制胶原的生成、平滑肌的迁移、增生，而减少内膜的增生。

Furthermore, preliminary work also performed to examine whether PI3K/AKT signal transduction pathway was activated in the process of refractory leukemia development. Materials and methods An immortalized human bone marrow stromal cell line, HS-5, was introduced to establish a bi-phase culture system for the cultivation of B-lineage precursor leukemia cells. ELISA and RT-PCR were used to investigate the expression of VEGF and its receptors in the leukemia cell lines and primary childhood leukemia cells in different treated groups. Flow cytometory method and immunofluorescent staining were employed to examine the apoptosis signals both in the VP16 treated and untreated leukemia cells. Western blot was utilized to explore the PI3K/AKT activated status in the drug induced or uninduced leukemia cells and lymphocytes from healthy donors.

材料和方法使用来源于人类骨髓基质细胞的细胞株HS-5作为滋养层细胞进行急性淋巴细胞性白血病细胞的体外培养，通过细胞生物学和免疫学方法评估培养体系并鉴定出难治性白血病细胞克隆；以ELISA和RT-PCR方法检测急性白血病细胞株和患儿白血病细胞VEGF及其受体的表达，了解不同治疗阶段VEGF及其受体的表达状况，并结合临床指标进行分析，明确VEGF及其受体在白血病发生过程中的作用；流式细胞仪和免疫荧光染色法对正常健康儿童、初发白血病患儿、复发白血病患儿及缓解后患儿进行凋亡因子检测和分析，初步阐明难治性白血病抗凋亡形成的原因；蛋白印记分析检测PI3K/AKT信号传导通路在健康儿童、初发白血病和复发白血病患儿的表达，初步了解难治性白血病形成的分子生物学机制。

objective:to investigate the expression and significance of rb in the occurrence, development and regression of infantile hemangiomas.methods:the expression of rb was examined in the proliferative stage and catagen of human hemangiomas and normal skin tissues by using immunohistochemical technique.immunohistochemical technique for factorⅷ-related antigen was used to prove that the cells which expressed rb were endothelium.image analysis system was applied to measure the expression level of rb at different stages of hemangiomas and in normal skin tissues.results:the expression of rb was significantly lower in proliferating hemangiomas than that in involuting hemangomas(p.05).conclusion:rb might play an important role in the regression of human hemangioma endothelial cells and anti-angiogenesis.

目的：探讨rb蛋白在血管瘤发生、发展及退化过程中的表达状况及其意义。方法：采用免疫组织化学方法检测人皮肤血管瘤增生期、退化期及正常皮肤组织中rb的表达水平，并结合第ⅷ因子相关抗原的免疫组织化学染色证实表达rb的细胞是血管内皮细胞。利用计算机图像分析技术测量不同时期血管瘤组织和正常皮肤组织rb表达的积分光密度和面积。结果：增生期血管瘤内皮细胞rb表达水平低于退化期，差异有显著性（p.05），退化期血管瘤内皮细胞rb表达水平与正常皮肤组织相比，差异有显著性（p.05）。结论：rb通过抑制血管瘤内皮细胞增殖和血管生成而在血管瘤的退化过程中起重要作用。

The correlation between Survivin and p53 expression and histological tyes and clinical stages in ovarian carcinoma were analysed.

结果： 1。卵巢癌p53和Survivin蛋白阳性表达率在卵巢癌组织中P53蛋白阳性表达率为54.84%(34/62)与正常对照组比较有显著差异(p＜0.01)；在浆液性囊腺癌中P53蛋白表达阳性表达率为56.52%(26/46)在黏液性囊腺癌中P53蛋白阳性表达率为57.14%(8/14)两组间无明显差异(p＞0.05)。

In this study, we have cloned and identified two full-length cDNAs of OsBIMK2 and OsBISERK1, which encoding MAPK and SERK, respectively, and associated with benzothiadiazole induced resistance.

本文克隆和鉴定了水稻中与苯并噻二唑(benzothiadiazole，BTH)诱导抗病性相关的编码MAPK的OsBIMK2基因和编码SERK的OsBISERK1基因全长cDNA；研究了OsBIMK2基因和OsBISERK1基因在水稻—稻瘟病菌亲和与非亲和互作过程中的表达模式；原核表达了OsBIMK2基因，获得重组OsBIMK2蛋白，并研究了重组OsBIMK2蛋白的生化特性；克隆了OsBIMK2基因的启动子序列，并运用农杆菌介导的瞬间表达系统分析鉴定了启动子中参与基因表达调控的可能顺式元件；将水稻OsBIMK2基因导入烟草，获得过量表达OsBIMK2基因的转基因植株，这些转基因植株中防卫反应基因PR-1组成型表达，并提高了抗病性。

If you are unfortunate enough to the lovelorn, please tell me, I will help you out, really, please contact me!

如果你不幸失恋了，请告诉我，我会帮助你摆脱困境，真的，请联系我啦！

China's plan to cut energy intensity by 20 percent and pollutant discharges by 10 percent between 2006 and 2010 is a case in point.

中国计划在2006年到2010间降低20%的能源强度和减少10%的主要污染物排放，就是一个这样的例子。