周质

P1 BMSCs, P3 CCs, and P3 ADAS were implanted into collagen/hyaluronic acid scaffold at different concentrations of 10%, 20%, 50%, 80%, and 100%, and the cells were then three-dimensionally cultured in serum-free culture media. Two weeks later, SO staining and immunohistochemical staining were employed to observe the synthesis between of chondroitin sulfate and type II collagen.

取P1骨髓间充质干细胞，P3软骨细胞和P3脂肪源性成熟基质细胞种植入不同比例(10%，20%，50%，80%，100%)的胶原/透明质酸支架中，在无血清培养液中三维培养。2周后，用SO染色和免疫组织化学染色分析，观察硫酸软骨素和Ⅱ型胶原合成情况。

Alternative etiologies for the development of this disorder include intrauterine insult (ischemic, metabolic, or viral such as CMV) during the 12th to 24th week of embryologic development. This results in damage to the germinal matrix and radial glial cells. Radial glial cells act as mechanical guides by which neurons are transported to the outer cortex during development.

另一种病因可能是由于第12-24周期间的宫内损伤（缺血、代谢异常、或病毒感染如CMV），其结果导致生发基质和放射状胶质细胞的损伤，在神经元移行至外皮层的过程中，放射状胶质细胞充当着机械性向导的作用。

In order to develop a safe and effective immunoadjuvant to enhance the immunity and resistance of animals against infection, a novel CpG Oligodeoxynucleotides containing 11 CpG motifs was synthesized and inserted into the VR1012 plasmid, designated as VR1C. Then the recombinant VR1C was entrapped with Chitosan nanoparticles prepared by the method of ionic cross linkage, and employed to inject muscularly 3-weeks old Kunming mice; the blank VR1012 packed with CNP and saline were used to inoculate mice as the control groups. 28 days after inoculation, all mice were orally fed with 0.4ml 2x108CFU/ per mouse virulent hemorrhagic enteritis E. coli to challenge the resistance against infection.

为研制安全高效免疫调节剂增强动物免疫抗病能力，本实验设计合成含11个C pG基序的寡聚核苷酸，重组构建含CpG的VR1012质粒(VR1C)；制备壳聚糖纳米颗粒包裹重组质粒(CNP-VR1C)，肌注接种3周龄昆明小白鼠，设壳聚糖包裹空质粒和生理盐水对照组；接种后28天口服大肠杆菌攻毒观察小鼠天然免疫的变化和对强毒感染的抵抗力，Sandwich ELISA测定血清免疫球蛋白和白细胞介素含量。

The PSCA_3 fragment was selected for its superior expression level in eukaryotic cells.Then the sig-PSCA_3-Fc-GPI genetic fragment was cloned into pVAX1-neo-IRES-GM/B7 vector to construct the final immunological inhanced DNA vaccine pVAX1-PSCA_3-FcGB. Immunofluorescence and flow cytometry were used to confirm the expression of PSCA_3 fragment by transfected into Cos7 cell.Finally,the anti-tumor effect of pVAX1-PSCA_3-FcGB was tested in murine prostate cancer model generated by RM-1 cell line.The animal was immunized with pVAX1-PSCA_3-FcGB DNA vaccine by intramuscular injection plus electroporation,pVAX1 and pVAX1-PSCA_1-FcGB plasmid were used as control.The inhibitory effect of tumor was investigated by observion of forming time,volume and inhibition ratio of tumor.Results:DNA sequencing conformed that the heterological PSCA fusion antigen fragment which was synchronized by overlapping-extending-PCR,was consistent to design.Enzyme digestion analysis showed that the 1 to 4 copies heterological PSCA fusion antigen fragments were constructed successfully.

方法(1)检索GenBank，选择包含人主要T细胞抗原表位序列的人PSCA基因片段，应用异种化抗原设计技术，保留人T细胞抗原表位，设计异种化PSCA融合抗原片段；(2)根据核酸序列按中心模板法设计引物，应用重叠延伸PCR技术拼接合成异种化PSCA融合抗原片段基因，以PCR、限制性酶切和DNA序列测定法进行鉴定：(3)利用DNA限制性内切酶BssHⅡ和MluⅠ酶切后粘端互补的特点，采用同尾酶法构建1—4拷贝异种化PSCA融合抗原片段(PNCA_1-PSCA_4)，并将上述片段分别插入真核表达载体pCI-neo-Fc-GPI中，转染293T细胞，借助免疫荧光+流式细胞术考察插入片段表达效率，最终选定PSCA_3片段进行下一步研究；(4)将sig-PSCA_3-Fc-GPI基因片段自pCI-PSCA_3-Fc-GPI质粒上切下，插入pVAX1-neo-IRES—GM/B7载体中，构建免疫增效DNA疫苗pVAX1-PSCA_3-FcGB，并应用转染Cos7细胞+免疫荧光/流式细胞术方法鉴定其在真核细胞中的表达情况；(5)给8周龄雄性C57BL/6小鼠皮下种植RM-1细胞，制备小鼠前列腺癌模型，并采用股四头肌肌肉注射+电脉冲法(Electroporation，EP)接种DNA疫苗质粒pVAX1-PSCA_3-FcGB，同时接种pVAX1空载体质粒和pVAX1-PSCA_1-FcGB质粒作为对照，通过观察计算免疫动物的成瘤时间、肿瘤体积和抑瘤率，来评价该DNA疫苗在小鼠体内的抑瘤效果。

We study the express of heparinase,matrix metalloproteinase MMP-2,MMP-14 and cadherin in the gastric cancer tissue and perigastric normal mucosal tissue,and make a statistics analysis between heparinase, matrix metalloproteinase MMP-2,MMP-14,E-cadherin and gastric cancer biological behavior.We hope to knom the possibility that heparinase,matrix metalloproteinase MMP-2,MMP-14,E-cadherin are regared as clinical screen index of the gastric cancer recrudescence or metastasis.

本文就肝素酶、基质金属蛋白酶MMP-2、MMP-14、钙粘蛋白在胃癌组织及胃周正常粘膜的表达进行了研究，并就肝素酶、基质金属蛋白酶MMP-2、MMP-14、钙粘蛋白与胃癌的生物学行为进行了统计学分析，以期了解肝素酶、基质金属蛋白酶MMP-2、MMP-14、钙粘蛋白作为胃癌复发或转移的临床筛选指标的可能性。

Methods DCs were prepared from peripherad blood mononuclear induce d with gra-nulocyt e/macrophage colony-stimulating factor and interleukin-4. Apoptosis of glioma cells were induced with γ-radiation. We design these experiment groups including (1) coculture of DCs and apoptotic glioma cells and T cells,(2) coculture of DCs and U937 cells and T cells,(3) coculture of DCs and cultured glioma cell and T cells,(4) coculture of Des and T cell.

用粒－巨噬细胞集落刺激因子加白介素－4(IL-4)从人外周血分化、诱导DCs、γ-射线在体外诱导培养的人脑胶质瘤细胞凋亡，将DCs、T淋巴细胞和凋亡胶质瘤细胞共培养，同时设计不同类型肿瘤细胞（U937及培养胶质瘤细胞）作对照，分离、富集DCs、T淋巴细胞进行免疫应答及肿瘤细胞杀伤试验。

When aborted rats in group E were treated with the dose of 50 IU/g IFN-γ, expression of TNF-αin this group was compared with the other four groups, the results were as follows: Compared with group A, the nucleus pre-opticus medialis, nucleus pre-opticus mango celluaris, nucleus periventricularis hypothalami, nucleus ventromedialis hypothalami, nucleus dorsomedialis hypothalami, nucleus arcuatus hypothalami and pars intermedia had a highly expression of TNF-α, in the other nuclei of hypothalamus, neurohypophsis and adenohypophysis, the change was not significantly, the expression of TNF-αin follicle and corpus luteum were decreased distinctly, the variation in uterus was slight and no difference was exsited; Compared with group B, the expression of TNF-αin nucleus nucleus lateralis hypothalami, nucleus ventromedialis hypothalami, nucleus arcuatus hypothalami, adenohypophysis and uterus presented a remarkable enhancement, conversely, the TNF-αexpression in the nucleus pre-opticus medialis, nucleus pre-opticus suprachiasmaticus, nucleus supra-opticus, nucleus pre-opticus mango celluaris, nucleus periventricularis hypothalami, nucleus paraventricularis hypothalami, neurohypophsis, follicle and corpus luteum had been significantly decreased; Compared with group C, the nucleus pre-opticus medialis, nucleus ventromedialis hypothalami, nucleus dorsomedialis hypothalami, nucleus arcuatus hypothalami, adenohypophysis, pars intermedia and uterus had upregulated the expression of TNF-α, however, it was decreased in the nucleus periventricularis hypothalami and corpus luteum; When compared with group D, the expression of TNF-αin nucleus lateralis hypothalami, nucleus dorsomedialis hypothalami, nucleus ventromedialis hypothalami, pituitary, follicle and ovary stroma were obviously enhanced, while in the nucleus supra-opticus and nucleus arcuatus hypothalami, the situation was definitely opposite, in the other parts, the change was not significant.

外源腹腔注射50 IU/g IFN-γ后，同A组相比：E组TNF-α免疫阳性产物在下丘脑视前内侧核、视前大细胞核、室周核、腹内侧核、背内侧核、弓状核、垂体中间部显著增高，在下丘脑其它核团、神经垂体、垂体前叶较A组差异不显著，在卵泡、黄体中阳性产物表达均较A组显著降低，在子宫中表达较A组变化不明显；同B组相比：E组TNF-α免疫阳性产物在下丘脑外侧核、腹内侧核、弓状核、垂体前叶、子宫均显著增高，在视前内侧核、视交叉上核、视上核、视前大细胞核、室周核、室旁核、神经垂体、卵泡、黄体中阳性物质表达均显著降低；同C组相比：E组下丘脑视前内侧核、腹内侧核、背内侧核、弓状核、垂体前叶与中间部、子宫均显著增高，在室周核、黄体中均显著降低；同D组相比：E组下丘脑外侧核、背内侧核、腹内侧核、垂体各部、卵泡、卵巢基质均显著增高，在视上核、弓状核显著降低，在其它部位表达变化差异不显著。

Results The articular cartilage constructed by the technique of centrifuge tube culture was hyaline cartilage.

结果在体外培养第2周时，组织切片可见软骨样组织结构；第3周时软骨发育趋于成熟，经阿新蓝染色基质丰富；第16周时，组织形态无明显变化，未见钙化。

Four weeks later, in the baicalin group, some spindle-shaped UCB-MSCs began to present shrinkage, with slender processes on the cell edge, and some UCB-MSCs tended to be spherical-, conical-, and triangle-shaped appearance, with many slender processes on the pseudopodia.

细胞形态学观察：脐血单个核细胞培养第2~3天开始贴壁生长，2周左右达高峰，3周后细胞生长达80%~90%融合，此时脐血原始间充质干细胞呈较均一的长梭形。4周后，黄芩苷组原来呈梭形的胞体一部分发生收缩，细胞边缘出现细的突起；一部分胞体逐渐近似圆形、锥形、三角形，伪足有多个细长突起。

It also had a pretest of object control skills in TGMD-2 (Test of Gross Motor Development 2nd Edition : including: Strike a Stationary Ball, Stationary Dribble, Catch, Kick, Overhead Throw, Underhand Roll), followed by another six weeks of lessons in games, at last there was a posttest a week after the lessons. This research analyzed the data by repeated measures t-test, and the level of significance was α=.05. Regarding quality, we promoted the teachers specialty through the process of designing games, teaching, practicing, self-examining and improving.

并於游戏课程介入前一周进行TGMD-2操作性动作技能的前测（包括定点击球、原地运球、踢球、接球、高手投掷、低手滚球），接著进行为期六周的游戏课程，且於课程结束后一周进行后测，研究所得资料以相依样本t考验进行统计分析，并将研究之统计显著水准介定在α=。05；在质的部份，藉由游戏教学设计、实践、反思、修正的历程，来提升教师之专业。

I can not make it blossom and suits me

我不能让树为我开花

When temperatures are above approximately 80 °C discolouration of the raceways or rolling elements is a frequent feature.

当温度高于 80 °C 左右时，滚道或滚动元件褪色是很常见的特征。