成组织细胞

Cumulative data have been documented that mesenchymal stem cells can differentiate into osteoblasts, chondrocytes, adipocytes, tenocytes, myotubes, neural-like cells, hematopoietic-supporting stroma and other non-hematopoietic cells derived from mesoblast or ectoblast.

大量研究表明，体外培养的骨髓间质干细胞在合适的诱导条件下，可分化为成骨细胞、成软骨细胞、脂肪细胞、成肌细胞、成腱细胞和神经元样细胞等多种非造血组织特别是中胚层和神经外胚层来源的组织细胞。

The palisade tissue cells of common type were 2 layers, while those of spur-type and columnar type were 3 or 4 layers. The ratio of palisade tissue and spongy tissue of common type was less than 1, and that of spur type and columnar type was more than 1. Moreover, the number of palisade tissue cell layers and the ratio of P/S were increased with the dwarfing degrees.

普通型苹果的叶片栅栏组织细胞一般排列成2层，其栅栏组织与海绵组织的厚度比值小于l；而柱型和短枝型的栅栏组织细胞通常排列成3层或4层，其栅栏组织与海绵组织的比值大于1，而且，随着矮化程度的加强，栅栏组织细胞层数和栅/海比有增加趋势。

Tumor cells grew actively in the tumor tissues of the control group. Prodrug therapy group: small amount of tumor cells were denatured vacuously,others were infiltrated by lympholeukocyte,and the growth of tumor cells were surspressed.Prodrug themochemotherapy group: what we can see that tumor cells were denatured vacuously, mesoplasts crinkled, cellular boundary disappered, only a small number of tumor cells remained, fibroblast were seen scatteredly, tumor cells were invaded by a lot of lympholeukocyte and eosinophilic granulocyte. Normal liver tissues, stomach tissues, lung tissues, pancreas tissues, small intestine tissues, large intestine tissues showed normal shape.

对照组肿瘤组织见肿瘤细胞生长活跃；前药治疗组肿瘤组织见有少量细胞空泡变性，少量淋巴细胞浸润，肿瘤细胞生长受到抑制；前药热疗组肿瘤组织可见肿瘤细胞空泡变性，细胞核皱缩，边集甚至消失，仅残留少量肿瘤细胞，并可见散在的成纤维细胞，大量淋巴细胞和嗜酸性粒细胞浸润；3组裸鼠正常肝组织、胃、肺、胰腺、小肠、大肠组织均呈正常形态学，无病理性损伤改变。

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。

Autologous MSC can be obtained easily,their source in human are plenty , own active proliferationand potential to difference into chondroblast.

自体骨髓基质干细胞来源丰富，取材容易，具有旺盛的增殖能力及向成软骨细胞分化的潜能，已成为软骨组织工程的首选种子细胞，但MSC诱导分化为成软骨细胞且应用于软骨组织工程的技术尚不十分完善，目前主要采用的技术为细胞团块离心管培养，其构建的软骨组织在力学性能上很难达到关节软骨的要求。

Autologous MSC can be obtained easily, their source in human are plenty, own active proliferation and potential to difference into chondroblast. Now MSC become the chief seed-cell in cartilage tissue engineer, but the technology to stimulate MSC differentiate into chondroblast is not very successful, the standard technology is culture the cell-ball in centrifuge tube, the reconstructured cartilage-like tissue is not achieve the mechanics of hyaline cartilage.

自体骨髓基质干细胞来源丰富，取材容易，具有旺盛的增殖能力及向成软骨细胞分化的潜能，已成为软骨组织工程的首选种子细胞，但MSC诱导分化为成软骨细胞且应用于软骨组织工程的技术尚不十分完善，目前主要采用的技术为细胞团块离心管培养，其构建的软骨组织在力学性能上很难达到关节软骨的要求。

Autologous MSC can be obtained easily,their source in human are plenty , own active proliferationand potential to difference into chondroblast. Now MSC become the chief seed-cell in cartilage tissue engineer , but the technology to stimulate MSC differentiate into chondroblast is not very successful,,the standard technology is culture the cell-ball in centrifuge tube,the reconstructured cartilage-like tissue is not achieve the mechanics of hyaline cartilage.

自体骨髓基质干细胞来源丰富，取材容易，具有旺盛的增殖能力及向成软骨细胞分化的潜能，已成为软骨组织工程的首选种子细胞，但MSC诱导分化为成软骨细胞且应用于软骨组织工程的技术尚不十分完善，目前主要采用的技术为细胞团块离心管培养，其构建的软骨组织在力学性能上很难达到关节软骨的要求。

Special staining methods, such as Masson and the Van Gieson staining were used to study the distribution of collogen fibers and elastic fibers. ResultsBy HE staining, the subepithelial connective tissues and vessels in the pterygium were more prominent than normal conjunctival tissues. An amorphous subepithelial superficial hyalinized zone and coarse eosinophilic granular materials were observed in the pterygia, but they were not found in normal conjunctival specimens. Coarse fibers were visible only in the deeper subepithelial connective tissues of pterygial samples. With Masson′s staining, the dense staining of collagen fibers was also more prominent in the pterygium than in the subepithelial connective tissues of normal conjunctiva. Abnormal collagen fibers were visible in the deeper sub-epithelial connective tissues of pterygial samples. With Van Gieson staining, abnormal collagen fibers were visible in the deeper subepithelial connective tissues. Dark coarse elastic fibers were found in the abnormal fibers only in the subepithelial deep connective tissues of pinguecula in the pterygia but not in the conjunctiva. With immunohistochemistry staining, MMP-3 was strong in the pterygial epithelium, moderate in fibroblast and absent from pterygial vascular walls. LN was strongly expressed in the blood vessel wall, moderately in the epithelial basement membrane and absent from the entire stroma.

结果HE染色：翼状胬肉组织上皮下基质中存在结缔组织的增生和血管形成；基质浅层存在一无定形物质透明区及粗糙的颗粒样嗜酸性物质，在翼状胬肉体部深层基质中存在粗糙的纤维组织；正常球结膜组织细胞排列整齐；基质为疏松结缔组织，胶原纤维平行排列，其间可见成纤维细胞，散在少量中性粒细胞、毛细血管；Masson染色：翼状胬肉浅层基质中存在致密的胶原纤维染色，深层基质中的胶原纤维存在变性样改变；VG染色：翼状胬肉组织深层基质中存在大量变性的胶原纤维，其间夹杂黑色的弹性纤维；免疫组化染色法：MMP-3在翼状胬肉上皮细胞中呈强表达，成纤维细胞中呈中等强度表达，血管内皮细胞中未见表达；LN在血管壁中呈强表达，在上皮细胞基底膜中呈中等强度表达，在整个基质中未见明显表达；col Ⅲ在整个翼状胬肉基质中呈强表达。

Objective: To explore the mechanisms resulting in the recurrence of urethral scar which make urethral strictures difficult to be cured, a series experiments were conducted to find potential effective factors involved in urethral scar formation and degradation, including the studies of extracellular matrix component of urethral stricture scar, the characteristics of urethral scar fibroblast, and the effects of urine on urethral fibroblast in vitro, as well as the studies to compare the difference of collagenase activity, type Ⅰ collagen and tissue inhibitor of metalloproteinase-1 (TIMP-1) in the tissues and cultured fibroblasts from normal urethra and strictured urethra respectively, and the studies to investigate the effect of antisense TIMP-1 oligodeoxyonucleotide on cell proliferation and collagenase activity of urethral scar fibroblast.

中文题名尿道瘢痕基础研究副题名胶原酶活性，TIMP-1的表达及其反义基因治疗外文题名 Experimental study on urethral scar-activity of collagenase，expression of TIMP-1，and antisense TIMP-1 gene transfection of urethral scar fibroblast 论文作者黄翔导师杨宇如教授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四川大学学位授予日期2002 论文页码总数104页关键词尿道手术瘢痕胶原酶成纤维细胞尿道瘢痕馆藏号BSLW /2003 /R699 /12 目的：研究尿道瘢痕的细胞外基质的组成，尿道瘢痕成纤维细胞的生物学特性以及尿液对其生长的影响；比较胶原酶活性，金属蛋白酶组织抑制因子-1（TIMP-1）以及Ⅰ型胶原含量在尿道瘢痕和正常尿道组织及体外培养的成纤维细胞中的差异；研究反义TIMP-1寡核苷酸对尿道瘢痕成纤维细胞增殖以及胶原酶活性的影响。

To understand the infectivity by porcine endogenous retrovirus with porcine skin fibroblast cell in vitro and in vivo, porcine skin fibroblast cell established by our laboratory were co-cultured with neo/HEK293 cell for the infection of RERV in vitro, and were subcutaneously transplantated to SCID (severe combined immuno-deficiency) mice for the infection of PERV in vivo, laying the foundation for valuation of biologic safety of xenotrans-plantation. The event of neo/HEK293 cells infected by PERV occurred during co-culture of porcine skin fibroblast cells with neo/HEK293 cells, expanding the rang of the infection of porcine endogenous retrovirus. Afterpig cells transplantated subcutaneously in SCID mice, the microchimerism (78.57%) of pig cells occurred widel, and there was phenomena of integration of PERV provirus (85.71%) in several organs or tissues remote from the injected sites, indicating infection of PERV in SCID mice in vivo. yet, there is no evidence of active viral replication in analysis of PERV env RNA of these tissues or organs.

为了解猪皮肤成纤维细胞PERV在体外和体内的感染性，通过建立猪皮肤成纤维细胞系，将所建细胞系与人胚胎肾293细胞体外共培养，并移植于严重联合免疫缺陷鼠皮下进行猪皮肤成纤维细胞PERV的体外和体内感染性实验，结果表明，猪皮肤成纤维细胞与人胚胎肾细胞共培养过程中，猪内源性逆转录病霉感染人胚胎肾细胞，进一步证实和拓宽了猪细胞PERV感染人细胞的范畴；猪皮肤成纤维细胞移植SCID鼠皮下后，导致SCID鼠发生猪细胞微嵌合(78.57%)和PERV在体内感染(85.71%)并且波及远离移植部位的多种组织或器官，但是并未检测出SCID鼠组织中表达PERV env RNA。

Cynanchum Lingtai apricot production in the average weight 65 grams, the brightly-colored fruit, juicy rich, sweet-sour taste, sweet from the nucleolus, when the late Qing Dynasty famous Shaanxi, Gansu provinces, the Qing imperial court Tongzhi tribute for years.

灵台生产的牛心杏平均单果重65克，果实色泽鲜艳，汁多味浓，甜酸适口，离核仁甜，清末时就驰名陕、甘两省，清同治年间曾为朝廷贡品。

Chenopodium album,Solanum nigrum, and Amaranthus retroflexus were very susceptible to the herbicides. Polygonum persicaria and Abutilon theophrasti were relatively less susceptible to the herbicides, and Lycopersicon esculentum was not susceptible to it. The relationship between reduction rates of weed biomass and PPM values of weed leaves 2,4, and 6 days after treatment was established.

供试的6种杂草对该混剂的敏感性存在显著差异：红心藜Chenopodium album、龙葵Solanum nigrum和反枝苋Amaranthus retroflexus对该混剂最敏感，ED90值分别为47.65、71.67和29.17g/hm2；春蓼Polygonum persicaria和苘麻Abutilon theophrasti敏感，ED90值分别为96.91、114.20g/hm2；而番茄不敏感。