后成质

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。

Materials and Methods: Six patients with left frontal lobe tumors (4 gliomas, 1 germinoma, 1 intra-axial cavernoma) close to Brocca and Wernicke language areas were studied. BOLD and DTI were performed on a 3.0T magnet GE Signa Excite~(? HD_X, BOLD using the word association paradigm developed in Part I and DTT technique developed in PartII. Language activation maps and fiber tracts were superimposed upon high resolution T1 weighted anatomic images of the brain for displaying the relationship between tumor, language functional areas and white matter tracts.

材料和方法：使用GE 3.0 T Singa Excite~ HD_x超导磁共振成像系统，对6例左侧语言相关脑区肿瘤(Ⅲ-Ⅳ级胶质瘤1例，Ⅱ级胶质瘤3例，生殖细胞瘤1例，海绵状血管瘤1例)行BOLD-fMRI及扩散张量成像扫描，BOLD应用语义相关判断作为任务刺激，所有数据进行离线后处理，应用AFNI软件处理BOLD图像，观察语言功能皮层的激活；DTT采用日本东京大学的Volume-one1.56和diffusion TENSOR Visualizer 1.5进行纤维束成像处理。

The location and structures of sex-pheromone-producing gland in female H.insularis were studied by EAG,GC,SEM,and TEM.These studies showed that thegland situate in the intersegmental membrane between the eighth and ninthabdominal segments,and is an eversible abdominal fold;Many plump cones disturbon the surface of the gland.The glandular cells of 2-day old virgin female H.insularis are arranged in one layer,among which the central cells are columnarepithelial cells and flat on two sides.The nucleus is irregular elliptical.There isevident conjugation between cells and the involution is more in the basal membraneof cell.Microvilli are distributed on the cytoplasmic membrane and linked withendocuticle on which there are many layers of chitin,and the outer cuticule is staineddeeper.The cell contains bubbles,mitochondria,glycogen deposits,roughendoplasmic reticulum and smooth endoplasmic reticulum.

结合触角电位、毛细管气相色谱、扫描电镜、透视电镜等技术对小线角木蠹蛾雌蛾腹尖末端不同组织部位提取物的测定分析以及腺体位置和形态结构的观察发现：小线角木蠹蛾性信息素分泌腺位于腹部末端8～9节之间，是一个由节间膜特化而成的上皮结构，为一可外翻的腹褶，腺体表面分布着饱满的锥形体，羽化后2天未交尾的雌蛾腺体细胞呈单层排列，腹面中央由密集的柱形上皮细胞组成，细胞排列向两侧延伸至背部，其形状由柱形逐渐变为扁平形，细胞核为椭圆形，细胞与细胞间有明显的胞连接，细胞基底膜基褶较多，质膜上分布着微绒毛，并与内表皮连接，内表皮上有多层几丁质，外角质层染色较深，细胞质中含有空泡、线粒体、脂质粒、粗面内质网和光面内质网。

Bone marrow-derived mesenchymal stem cells are capable of chondrogenesis, making them a possible source of cells for injectable cartilage tissue engineering. There exist different ideas on the ability of mesenchymal stem cells's chondrogenesis in monolayer culture. Because of this, the effect of adult rabbit's bone marrow-derived mesenchymal stem cells chondrogenesis in monolayer culture was studied. The mesenchymal stem cells was isolated from adult rabbit's bone marrow and monolayer cultured. TGF-β1, Vit-C and Dexamethasone were used. Immunohistochemistry analyses and histological staining of H-E, Methylaniline blue and Alcian blue were performed to identify the expression of collagen type Ⅱ and cartilage associated matrix. The results showed that the induced cells expressed and produced collagen type Ⅱ and cartilage associated matrix. This suggests that the differentiation of adult rabbit's marrow-derived mesenchymal stem cells into chondrocyte in monolayer culture is feasible and may be induced by TGF-β1, Vit-C and Dexamethasone.

骨髓基质干细胞的软骨分化潜能使其可能成为可注射组织工程化软骨研究的种子细胞，为探讨体外培养的骨髓基质干细胞在平面诱导条件下软骨分化的可行性，我们进行下列实验：获取并体外平面培养成体兔骨髓基质干细胞，应用TGF-β〓、Vit-C和地塞米松对其软骨分化诱导，诱导后的骨髓基质干细胞行细胞爬片组织学和Ⅱ型胶原免疫组化，结果证实，诱导后骨髓基质干细胞可分泌Ⅱ型胶原，组织学染色可见类似于软骨细胞，由此证明体外培养的骨髓基质干细胞在平面诱导条件下可以软骨分化，其软骨诱导因子为TGF-β〓、Vit-C和地塞米松。

Results Infected by this peptide, cell viability decreased 28.9%. Under light microscope, cells were shrinked and rounded, many cells were divorced from plate wall, some neuraxon shortened and broke. Apoptotic cells which nucleolus shrinked and rounded could be coloured orange by fluorescent colouration. Under electron microscope, chromatin gathered along the inside of the nuclear membrane, vacuole bodies appeared. Apoptotic peak was detected by flow cytometry and the ladder band appeared in DNA electrophoresis.

结果：细胞接触肽段后存活率下降28.9％；光镜下可见细胞贴壁不良，胞体缩小，细胞突起断裂缩短；荧光染色可见细胞突起缩短、胞核固缩、胞质染成橘红色的凋亡细胞；电镜下可见胞质中出现空泡样结构，细胞染色质浓集于核膜内侧并裂解成碎块状；流式细胞仪检测细胞出现亚二倍体峰，DNA电泳出现梯状带。

They are most abundant in quartz of pegmatitic leucosomes and granitic rocks, and represent the fluid appearing in their cooling stage. The DL(H2O) of such fluid is 0.93～0.96g/cm3 corresponding to P≈0.6GPa which is compatible with condition when the rapid decompression in this region was ended. 3 CO2-H2O two or three phases inclusions. They are the most widespread, and more frequently in clusters and in intragranular tails in quartz. The relative content of CO2+CH4 and H2O is considerably variable. They are also characterized with lower density (0.6～0.8g/cm3) and low pressure about 0.3～0.4GPa, and may be originated by mixture of carbonic liquid from breaking of most inclusions of peak stage and aqueous liquid of magmatic source. After entrapment, the further decrease of temperature to lower than 330℃ made such fluid separated to two or three phases.

主要集中于伟晶质脉体和花岗岩中，是这期岩浆冷凝过程析出的流体相当时就被封闭所成，密度为0.93～0.96g/cm3，相应压力约0.6GPa，这与峰期后迅速减压过程结束时的条件相符。3CO2-H2O两相和多相包体，分布最广，成簇状和拖尾状包体群，碳质和H2O相对含量很不均匀，整体密度相对最低，一般为0.6～0.8g/cm3，相应压力为0.3～0.4GPa，它们可能是大幅度减压过程中第一类包体大量爆裂析出的碳质流体与第二类H2O溶液流体在各处以不同比例混溶所成，它们被封闭后在降温至330℃以下时分裂成两相或三相包体。

The results indicated that the typical apoptosis was induced by PRRSV in lungs and uteri. The ultrastructural changes showed different characterizations at different stages of the infection. During the early stage from 8th h to day 3 postinfection,the apoptosis cells shrank,the cytoplasm concentrated,the chromatin condensed,the kernel disaggregated and the endoplasmic reticulum dilated. During the middle stage from day 5 to day 9 postinfection,the apoptosis cells evidently shrank,the cell membranes protu berated,the plastosome manifolded,and apoptosis bodies were found. During the last stage from day 10 upwards postinfection,the apoptosis bodies degenerated and disappeared,and no inflammation occurred. Few apoptosis cells were necrotic.

结果表明，PRRSV可诱导宿主肺和子宫发生典型的细胞凋亡，表现为细胞的超微形态结构随着病毒感染进程出现不同的特征性变化，早期（感染后第8 h至第3 d）凋亡细胞多表现为细胞体积缩小，细胞质密度增强，染色质浓缩，核仁解体，内质网扩张；中期（感染后第5 d至第9 d）凋亡细胞体积显著缩小，细胞膜突起，线粒体增多，有凋亡小体形成；晚期（感染后第10 d以后）凋亡细胞形态多表现为凋亡小体降解和消失，少数凋亡细胞在凋亡晚期表现为坏死细胞。

Results The mortality rate of mice in 80 mg/kg, day cyclophosphamide group was 16.7%, and T level [ at 30th day :( 1.38 ± 0.31 );45th day:( 1.15 ± 0.26 ) ] and T/LH ratio [ at 30th day:(0.163 ± 0.014); 45th day:(0.127 ± 0.023 ) ] were significantly decreased (all P＜0.05) at 30th day after induction;The concentration of MDA [at 15th day:(2.70 ± 0.41);30th day:(2.710.36);45th day:(2.67 ±0.43) ] was maintained at a high level (all P＜0.05) during the 45 days ; Number of Leydig's cells [ at 15th day:(9.65 ± 0.75 ); 30th day:( 14.05 ± 0.67 ); 45th day:(8.49 ± 072)] and layers of spermatogenetic epithelia [ at 15th day:(4.75 ± 0.82);30th day:(3.60 ± 0.49);45th day:(3.74 ± 0.43 ) ] were significantly decreased ( all P ＜ 0.01 ) and stabilized in a low level. The induced model was stable and the mortality rate was acceptable. In the 60 mg/kg, day cyelophosphamide group, the T level and T/LH ratio had no significant change (P ＞ 0.05 ), and the concentration of MDA ,number of Leydig' s cell and layers of spermatogenetic epithelia recovered at 30th day after induction. The induced model was unstable.

结果 剂量每日为80 mg/kg体重小鼠成模后死亡率为16.7%，血清T[30 d：(1.38±0.31)；45 d：(1.15±0.26)]及T/LH比值[30 d：(0.163±0.014)；45 d：(0.127±0.023)]于诱导后第30天出现显著下降(P均＜0.05)，而诱导后睾丸组织内MDA含量[15 d：(2.70±0.41)；30 d(2.71±0.36)；45 d：(2.67±0.43)]维持高水平(P均＜0.05)，生精上皮层次[15 d：(4.75±0.82)；30 d：(3.60±0.49)；45 d：(3.74±0.43)]和间质细胞[15 d：(9.65±0.75)；30 d：(14.05±0.67)；45 d：(8.49±0.72)]均显著减少(P均＜0.01)并稳定于低水平，模型稳定，死亡率适当；每日60mg/kg体重组小鼠血清T及T/LH比值于不同时段并未出现明显变化(P＞0.05)，且睾丸组织内MDA含量、生精上皮层次和间质细胞计数在30 d后有所恢复，模型不稳定；每日100 rag/ks体重组死亡率为30.0%，死亡率过高。

Pristine Li-Al LDHs are synthesized by hydrothermal process in different reaction conditions by varying the aging time for 1 and 24 hours. Both the Li-Al layered double hydroxides (abbreviated as Li-Al LDH1 and Li-Al LDH24 for aging time 1 hour and 24 hours, respectively) were modified by using sulphanilic acid sodium salt hydrate, modified agent to form modified Li-Al LDH1-SAS and Li-Al LDH24-SAS. The morphology of the pristine layered double hydroxides are investigated by using wide angle X-rays diffraction, scanning electron microscopy and particle size analyzer which indicates that the crystallinity and aspect ratio is increased with increasing the aging time from 1 hour to 24 hours. The particle size of Li-Al LDH24 and Li-Al LDH1 are found to be ~1000 nm and ~250 nm, respectively. Both the pristine and modified LDHs are characterized by XRD, FTIR spectra and thermogravimetric analysis.

本研究目的在於改变长晶时间的长短，合成出同径大小的Li-Al LDHs，再经由改质剂sulphanilic acid sodium salt hydrate将Li-Al LDHs无机层材表面有机官能化制备出改质型Li-Al LDHs-SAS，於是进一步藉由扫描式电子显微镜、射径仪和穿透式电子显微镜等仪器分别观察LDHs改质前和改质后其主层结构型态上的变化；从中可以发现Li-Al LDHs随著长晶时间的增加其径有随之增大的趋势，但经有机化改质后其径会由於改质环境的影响明显低许多，针对此现象本实验将未改质和改质后之Li-Al人工无机层材制备成复材进一步探讨其在热性质和难燃特性上的为表现。

METHODS: BMSCs were obtained from Japanese big-eared rabbits, and in vitro cultured. Then the subcultured BMSCs were transfected by pCDNA3.1 plasmid, followed by incubation on swine SIS to construct the tissue-engineered skin. The growth of cells and phenotype of BMSCs were detected by flow cytometry.

日本大耳兔骨髓间充质干细胞进行体外培养扩增后，通过pCDNA3.1质粒将碱性成纤维细胞生长因子基因转染至生长状态良好的骨髓间充质干细胞，并将转染后的细胞接种于制备好的猪小肠黏膜下层上，进行体外联合培养，构建组织工程皮肤。

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。