后成质

Results Compared to those before treatment the volume of the implanted keloid of Group D began to decrease since 14 days after treatment time-dependently (all P 《 0.05), and the volumes of the other 3 groups continued to increase and peaked on days 21, 14, or 7 respectively (all P 《 0.05).Microscopy showed infiltration of a larger quantity of histiocyte in the keloid tissue, and more obvious collagen disorganization and apoptosis of fibroblasts in Group D than in the other 3 groups.The protein expression of Bcl-2 was more remarkable and the protein expression of BAX was less remarkable in Group D than iu the other 3 groups.

结果 用药前和用药后7、14、21、28、35 d，D组瘢痕疙瘩组织块体积(mm3)分别为173±5、172±5、147±5、125±6、112±7和84±9，从用药后14 d开始明显缩小(均P＜0.05)；而其他3组瘢痕疙瘩组织块体积均明显增大，从用约后7 d开始各时点测得的体积均明显大于D组(均P＜0.05)。D组瘢痕疙瘩组织中有大量小鼠组织细胞浸润，胶原结构破坏和成纤维细胞凋亡明显重于其他3组，Bcl-2蛋门质表达明显弱于而Bax蛋白质表达明显强于其他3组。

On the basis of our results,IFN-θnot only has the sequence homology to other typeⅠinterferons,but also matches the primary properties of typeⅠinterferons in antiviral, IFNαR receptor binding and virus inducing,etc.

首先，将PoIFN-α基因克隆入pcDNA3真核表达载体中，筛选阳性克隆后大量抽提质粒，通过脱水化-再水化步骤将重组质粒包裹入阳离子脂质体内制备成核酸佐剂(IFN-adjuvant)。

RESULTS: Twelve days after mandibular vertical functional positioning the condylar height increased. Nine days after mandibular vertical functional positioning the thickness of prechondroblast layer and chondroblast layer increased significantly compared with control groups. However the thickness of hypertrophic layer decreased significantly after 6 days' mandibular repositioning. No change was found in the thickness of mesenchymal layer during the experimental period.

结果：下颌垂直向功能性移位后12d，髁突高度较对照组显著增加，前斜面更倾斜；髁突后上区前成软骨细胞及成软骨细胞层厚度在实验第3~6天无明显差异，第9天开始出现显著变化，实验组较对照组显著增厚，这种变化持续到实验第12天；肥大软骨细胞层厚度在实验第3、9、12天，实验组与对照组无显著差异，在实验第6天较对照组显著减少，间充质细胞层变化不大。

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。

MAIN OUTCOME MEASURES: Morphological changes in primary cultured and subcultured mouse BMSCs were measured. BMSC surface antigen of the second passage of mice (CD105, CD44, CD25, CD34) were determined using flow cytometry. The modified method was assessed to harvest the purity of stem cells. Activity of mouse BMSCs was identified using adipogenic and osteoplastic differentiation. The strength of fluorescent cells following multiple passage was observed.

主要观察指标：原代及传代小鼠骨髓间充质干细胞形态变化；对第2代小鼠骨髓间充质干细胞表面抗原，CD105，CD44，CD25，CD34进行流式细胞仪检测，评价改良方法获取干细胞纯度；通过成脂成骨分化，鉴定小鼠骨髓间充质干细胞的活性；观察多次传代后荧光细胞强度。

The human cartilages are composed of chondrocyte and extracellular matrix , the form of chondrocytes are hypertrophy and the quantity are less; the ECM of cartilage are compised of type Ⅱ collagen and proteoglycan. Articular cartilages are all hyaline with little fibers. Trauma and arthritis are the main cause of cartilage injury, the ommilayer injury ofcartilage can be recovered by marrow, but because of without stimulation mechanism, the new tissues are merely fibrocartilages, they can not be coincide with hyaline cartilage in menchanics; the purely damage of articular cartilage can not stimulate chondrocyte to regenerate because of without blood circulation, thus, the plerosis of articular catilage can not depend on the proliferation of local chondrocyte. Ever since, people tried their best to find a way to reconstruct articular cartilage.

中文题名人骨髓基质干细胞成软骨诱导及多孔复合材料作为细胞载体的体外实验研究副题名外文题名 Cartilage induction of human mesenchymal stem cells and experiment on compound porous materials as cells' scaffold in vitro 论文作者刘晓岚导师周江南学科专业外科学研究领域\研究方向学位级别博士学位授予单位中南大学学位授予日期2003 论文页码总数68页关键词骨组织工程软骨细胞骨髓基质干细胞壳聚糖高分子外消旋聚乳酸馆藏号BSLW /2003 /R68 /10 造成人体关节软骨损伤的原因主要为创伤和关节炎，关节软骨全层损伤可由于骨髓中间充质干细胞的高速增殖修复，但这种修复由于缺乏相应的刺激机制，只能形成纤维软骨，而不能形成符合关节生理、力学要求的透明软骨；单纯软骨部分损伤软骨组织内无血管，软骨细胞迁移迟缓，无法使损伤区域软骨细胞再生，因此，关节炎及关节创伤后的软骨修复不能依赖于软骨细胞的增殖和迁移。

In order to enhance the miscibility between the inorganic and the organic phases,the silanol coupling agent is made from the slane VTMS and the colloid silica. Then the silanol coupling agent and nano-particles of ZnSb2O6 as the conductive filler are successfully bonded and the nano-particles of ZnSb2O6 will become more organic-like. The colloid silane-SiO2-ZnSb2O6 precursor solution is subsequently mixed with a homogeneous lsopropylalcohol solution of SR-399 acrylic monomers (Dipentaerythritol pentaacrylate) and the active free radical agent709. Under nitrogen purging, this reaction solution prepolymerize at 60℃. Then, the solution is spin-coated with the spin coator at room temperature. At last, the coated thin film is cured and crosslinked by the UV curing reactor to form the highly transparent thin film with good optoelectronic properties from hybrid organic-inorganic nano-composite.

首先以矽氧烷VTMS和colloid silica制备矽氧偶合剂，再以此偶合剂对分散於异丙醇溶液之奈米级的colloid ZnSb2O6做有机化改质，以增进其与有机高分子基材的键结相容性，我们将此colloid silane-SiO2 - ZnSb2O6前驱物与SR-399有机基材单体(Dipentaerythritol pentaacrylate)的异丙醇溶液与活性自由基光起始剂709均匀掺混后，在充满氮气反应系统下，加热至60℃进行预聚合反应，接著在室温下以旋转涂布机旋转涂布成厚度均匀薄膜，最后将此薄膜直接高温硬化或以UV硬化机网状键结硬化成透明与具部份导电性等光电特性之无机/有机奈米混成薄膜。

Methods:Human CD80 cDNA was transfected into B16 melanoma cells by lipofectin-mediated gene transfer before the expansion of tumor cells were tested by MTT method in vitro;C57BL/6 mice were inculated subcutaneously either mock-transfected B16 cells (B16-neo)or CD80-transfected B16 cells (B16-CD80),then the latency,survival times and tumor mass growth were investigated.The lymphocytes were examined for both proliferation indices and specific cytotoxic activity by MTT method and improved LDH-releasing method,after syngenic Mixed Lymphocyte tumor cultures.

通过脂质体介导将CD80基因导入小鼠黑色素瘤B16细胞后，利用SABC法检测CD80的表达；MTT法测定肿瘤细胞体外增殖能力；将肿瘤细胞接种至同源小鼠皮下后观察成瘤期、荷瘤小鼠存活期及肿瘤结节生长速度；在同源淋巴细胞肿瘤细胞混合培养后，通过测定淋巴细胞增殖指数和CTL杀伤活性检测肿瘤细胞的免疫原性。

Damage hair is qualitative must first daub of the ability after attenuant emollient , also can allocate with 3 agents, scale chooses according to the damage level of the hair, if discover after bate judgement error brings about bate excessive should pour water instantly, develop the 3rd agent, if the hair is sere only OK redo, if the hair has resembled cotton shrink into oneself is washed-up.

受损发质一定要先稀释软化剂后才涂抹，也可以用三剂调配，比例则根据头发的受损程度来选择，如在软化后发现判断失误导致软化过度则应立即冲水，冲水后涂抹上第三剂，如头发只是干枯则可以再做，如头发已经像棉花一样缩成一团就不行了。

Nerve and ligament tissues could be clearly observed from light and soft patch, and this might and prevent hurt by mistake, improve repairing operability, enhance correct putting of patch, and decrease incidence of pain.

超薄轻质补片更柔软、更透明、操作容易，透过网片能清楚地观察到网片后方的神经和韧带等组织，可预防误伤，提高了修补中的易操作性，并且便于网片的正确放置，减少了修补后疼痛的发生率，而且网片的结构更有利于成纤维组织的长入，能很好地适应患者修补后因体位变动和肌肉收缩而产生的纵向和横向移动，患者修补后的不适感也极大地降低等优点。

The split between the two groups can hardly be papered over.

这两个团体间的分歧难以掩饰。

This approach not only encourages a greater number of responses, but minimizes the likelihood of stale groupthink.

这种做法不仅鼓励了更多的反应，而且减少跟风的可能性。