糖质的

ConclusionA highiron animal model can be established by intraperitoneal injection of iron dextran. The changes of various parameters result in damage to the organs, it is possible that the ferri ions were released through metabolism of iron dextran and freeion mediates lipid peroxidation to cause the damage.

给小鼠腹腔注射右旋糖酐铁，可以成功建立高铁动物模型，其各项指标有明显变化并对各脏器造成损害，其机制可能与右旋糖酐铁在体内代谢释放出铁离子，游离的铁离子介导脂质过氧化而造成的损害等有关。

ConclusionA highiron animal model can be established by intraperitoneal injection of iron dextran. The changes of various parameters result in damage to the organs, it is possible that the ferri ions were released through metabolism of iron dextran and freeion mediates lipid peroxidation to cause the damage.［KEY WORDS］IronDextran complex; Models, animal; Lipid peroxidation

给小鼠腹腔注射右旋糖酐铁，可以成功建立高铁动物模型，其各项指标有明显变化并对各脏器造成损害，其机制可能与右旋糖酐铁在体内代谢释放出铁离子，游离的铁离子介导脂质过氧化而造成的损害等有关。

G. biomimetic modification of biomaterials surface for improving cells affinity, carbohydrate cluster, glycolipid and protein modifying biomaterials surface in order to enhance cells specificity recognition, self2assembly of materials to modify surface topography1

基于生物材料对细胞生长的影响，本文提出了生物材料表面生物仿生化以提高细胞亲和力，糖链团簇、糖脂质及材料表面蛋白质修饰以提高细胞特异性识别，材料表面的自组装修饰以改善表面形态等观点。

Many glycoproteins of lower and higher eucaryotes are attached to the plasma membrane by means of a glycosylphosphatidylinositol .

很多高等和低等真核生物的糖蛋白是以糖基磷脂酰肌醇锚定的方式被连接在质膜上。

Many glycoproteins of lower and higher eucaryotes are attached to the plasma membrane by means of a glycosylphosp...

很多高等和低等真核生物的糖蛋白是以糖基磷脂酰肌醇锚定的方式被连接在质膜上。

The colony positive rate exam which after the trisaccharide iron urea indol culture medium is 28.91%,higher than the exam the colony positive rate using the trisaccharide iron culture medium (7.67%).

经三糖铁尿素靛基质培养基筛检出的菌落阳性率为28.91%，比三糖铁培养基筛检出的菌落阳性率（7.67%）高。

The study showed the recombinant wt/mCREG protein depressed the VSMC proliferation depending on dose and the optimal concentration was 400nM;2biologic function of CREG protein and the membrance receptor mechanism:①effect on VSMC migration: the wound healing experiment showed the OB2 cells migration was slower significantly after added wt/mCREG(400Nm) in supernatant. The HITASY cells migration were very slowly and no remarkable change. The gelatinase digestion and Western blot analysis showed the matrix metalloproteinase was decreased and TIMPs was increased;②effect on differentiation: after added wt/mCREG(400nM), the expression of myocardin, SMα-actin, MHC and caldesmin were increased and that of LM-1 and FN were decreased in OB2 cells. These effects were more significant when adding wtCREG.;③effect on VSMC proliferation: Cell cycle assay and BrDU stain showed: after added the wtCREG and mCREG protein, the ratio of cell in G0/G1 phase increased to 0.5773 and 0.5572 from 0.5308 respectively in OB2 group, which increased to 0.7369 and 0.7034 respectively from 0.6297 in HITASY group;3Role of M6P/IGF2R in CREG biologic function:①ELISA and co-immunoprecipitation showed the wt/mCREG binding to M6P/IGF2R directly.②antibody blocking test: when the anti-IGF2R was added to medium at the same time with wt/mCREG at different concentration(2μg/mL、4μg/mL、8μg/mL),the effects of CREG protein which depressing proliferation, migration, secretion and promoting differentiation were blocked, which had the positive correlation to the concentration of added anti body. The studies showed two combinant CREG promoted VSMC switch to differentiation phaenotype, at the same time, depress VSMC proliferation, migration and secreting extracellular matrix.

上述实验结果证实：两种重组CREG蛋白对VSMC增殖均有剂量依赖性的抑制作用，并且相同浓度的糖基化的CREG蛋白对细胞增殖的抑制效应更为显著，最佳效应浓度为400nM；2两种重组CREG蛋白添加后对HITASY和OB2细胞生物学行为的影响：①CREG蛋白对VSMC迁移的影响：刮伤实验发现，加入最佳效应浓度的wtCREG和mCREG蛋白24h后，OB2组迁移能力下降，HITASY组无明显变化；细胞外基质金属蛋白酶-2，9(Matrix metallo-proteinase 2，9，MMP2 ，9)明胶酶电泳检测和Western blot检测结果证实，两种CREG蛋白均可以使OB2细胞合成细胞外基质MMP2，9减少，而组织金属蛋白酶抑制物(Tissue Inhibitors of Metalloproteinases，TIMPs)增加；②CREG蛋白对VSMC分化的影响：加入400nM的wtCREG和mCREG蛋白12h后，OB2细胞myocardin、SMα-actin、MHC、caldesmin表达增加，LM-1、FN表达减少；③流式细胞仪分析细胞周期和BrDU染色分析证实，加入400nM的wtCREG和mCREG蛋白后，OB2组G0/G1期细胞由0.5308分别增加至0.5773和0.5572，HITASY组G0/G1期细胞由0.6297分别增加至0.7369和0.7034；3M6P/IGF2R在重组CREG蛋白的生物学功能中的调控作用：①免疫共沉淀和免疫荧光双染色分析结果显示，CREG蛋白与M6P/IGF2R存在直接结合；②应用抗体阻断实验：将不同浓度的anti- M6P/IGF2R(2、4、8μg /mL)与两种CREG蛋白同时加入培养液中，CREG蛋白抑制VSMC增值、迁移和合成细胞外基质、促进分化的效应减弱，而且与加入anti- M6P/IGF2R浓度正相关。

Endocellular enzymes of Arthrobacter nicotinovorus D-97 showed a weak hydrolytic activity towards α-1, 4 linkage and glucose, maltose, sucrose and lactose could not be converted into trehalose.

综合采用薄层层析、高效液相电喷雾电离质谱联用(HPLC-ESI／MS)及核磁共振分析手段，在酶反应产物不必被纯化的前提下，最终证明食尼古丁节杆菌D-97对麦芽寡糖的酶反应产物中含有海藻糖。

After being digested with EcoR I and Xho I, they were inserted to the vectors pGEX-4T-l orientally, and then transformed into E.coli BL21(DE3) cells. The monoclone were selected and identified by means of restriction analysis, PCR and sequencing. As the result, the two recombinant plasmids with the different DNA fragements of the PrP gene were constructed and designated as pMY01[Ov rPrP(23-244)] and pMY02[Ov rPrP(91 -244)], respectively.

挑选单个菌落，提取重组质粒，经酶切、PCR扩增后琼脂糖凝胶电泳分析和测序鉴定，成功地构建了2种绵羊重组朊蛋白的表达质粒pMY01[OvrPrP(23～244)]和pMYO2[Ov rPrP(91～244)]。

The PCR products were examined by agarose gel electrophoresis. The target gene fragments were purified by gel extraction kit and ligated to cloning vector pMD18-T. The recombinant vectors were transformed into host strain E. coli K802 by lithium chloride method, screened and identified with PCR and restrictive enzymatic digestion. Their sequences were confirmed by DNA sequencing.(2) sTWEAK1 gene was subcloned into expression vector pProEx HTb and transformed into E. coli BL21. sTWEAK2 gene was subcloned into expression vector pMAL-C2x and transformed into E. coli TB1. The recombinant vectors were screened and identified with PCR and restrictive enzymatic digestion. The recombinant fusion proteins were induced to express with IPTG, detected by coomassie brilliant blue-stained SDS-polyacrylamide gel electrophoresis , and confirmed by Western blot analysis.(3) The sTWEAK1 fusion protein was purified with Ni-NTA Spin Kit.(4) The biological activity was assayed on transformed and tumor cells by microplate photometer after crystal violet or sulfur rodamine B staining.(5) The contents of IL-8 in the supernatant of 1990 cell cultures were determined by ELISA.(6) The morphological changes of the sensitive cells were observed by light and transmission electron microscopies.(7) The cell cycle and apoptotic rate were assayed by flow cytometry in 1990 and M85 cells.(8) The effect of fusion proteins on induction of NF-κB in 1990 and LOVO cells was detected with Dual-Luciferase Reporter Assay system.(9) The TWEAK gene was subcloned into Adeno-X Viral DNA with pShuttle vector and transfected into HEK293 cells by lipofectamine method.

(1)本研究用RT-PCR方法，从人组织细胞总RNA中扩增可溶性TWEAK胞外区（sTWEAK1和sTWEAK2）的cDNA序列及全长编码序列，用琼脂糖凝胶电泳分析PCR产物，胶回收目的基因片段，连接到pMD18-T克隆载体中，转化大肠杆菌K802，PCR和酶切筛选阳性克隆，全自动DNA测序验证序列；(2)sTWEAK1和sTWEAK2分别亚克隆到pProEx HTb和pMAL-C2x表达载体中，分别转化大肠杆菌BL21和TB1，PCR筛选和酶切鉴定，阳性克隆用IPTG诱导表达，表达产物用SDS-PAGE分析和Western blot验证融合蛋白；(3)用NTA-Ni Spin试剂盒初步分离纯化sTWEAK1融合蛋白；(4)用体外培养的肿瘤细胞和正常对表达产物进行活性检测，贴壁细胞用结晶紫染色法，悬浮细胞用磺酰罗丹明B染色法，酶标仪检测OD值；(5)敏感细胞用ELISA法检测细胞培养上清中IL-8的含量；(6)用光镜和电镜观察敏感细胞死亡和细胞凋亡情况；(7)用流式细胞仪分析表达产物对敏感细胞凋亡率和细胞周期的影响；(8)用双荧光素酶报告基因检测法，测定表达产物对敏感细胞NF-κB的影响；(9)用pShuttle穿梭质粒将TWEAK重组到腺病毒载体上，用脂质体转染法转染HEK293细胞，PCR鉴定重组质粒。

For a big chunk of credit-card losses; the number of filings (and thus charge-off rates) would be rising again, whether

年美国个人破产法的一个改动使得破产登记急速下降，而后引起了信用卡大规模的亏损。

Eph. 4:23 And that you be renewed in the spirit of your mind

弗四23 而在你们心思的灵里得以更新