成纤维细胞的

Methods: CF were isolated and cultured from Neonatal SD rats.The cells were divided into different groups and treated with Ang-(1-7), TGF-β1, TGF-β1+ Ang-(1-7), and the acceptor inhibiter of TGF-β1 respectively.

分离培养SD仔鼠心脏成纤维细胞，以Ang-(1-7)、TGF-β1、Ang-(1-7)+TGF-β1组、TGF-βⅠ型受体抑制剂等干预，通过WST-1法检测细胞增殖，RT-PCR测定心脏成纤维细胞Smad3、Smad 7mRNA的表达。

AFGF shows greatpotential for clinical applications for therapy of a variety of diseases, such as Parkinson'sdisease, spinal cord contusion injury, neural regeneration in reimplantation of broken-offfinger, brain ischemia, renal ischemia, myocardial infarction, occlusive vascularitis, retinalischemia, gastric ulcer and nonhealing wound and so on. We expressed human acid fibroblast growth factor gene transiently in tobacco byAgro-bacterium mediated vacuum infiltration and studied the factors, which influenced theexpression level.

人类酸性成纤维细胞生长因子(aFGF，FGF-1)是成纤维细胞生长因子超家族成员，来自于三个胚层的多种细胞都可以表达aFGF.aFGF在治疗帕金森综合症、急性脊柱扭曲性损伤、断指中神经功能重建、脑缺血、肾缺血、心肌梗塞、闭塞性脉管炎、视网膜缺血、胃溃疡及难愈合性伤口等多种临床应用方面具有巨大潜力。

Methods:(1) ADM was produced from swine skins treated with trypsin followed by Triton X-100.(2) type I collagen of mouse tail was extracted by 0.5mol/L acetoacetic acid.(3) Two kinds of ADM were got by being soaked with 0.02% glutaraldehyde for 5~10 min or being soaked with mouse type I collagen for 24h, then preserved at 4℃.

(1)应用胰蛋白酶消化-去污剂法制备ADM；(2)用0.5mol/L乙酸溶液提取制备I型鼠尾胶原；(3)用0.02%戊二醛溶液浸泡ADM5-10min，制成交联型ADM；另一部分ADM用质量分数0.25%的I型鼠尾胶原浸泡24h，制成胶原包埋型ADM，冰冻保存；(4)用酶消化法培养原代成纤维细胞，取第三代增殖期细胞，将其调整至浓度为2×105/ml的成纤维细胞悬液。

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。

AIM: To investigate the effects of dibairen decoctum on the proliferation of keratinocytes and fibroblasts as well as the potential effects of DBRD on fibroblast contraction.

目的：观察地白忍煎剂对角朊细胞和成纤维细胞增殖的影响以及对成纤维细胞收缩功能的潜在作用。

By the cooperation of 8-MOP/UVA, the photoaging-characteristic biological markers of the dermic fibroblasts in vivo and in intro were changed as follows:① with a permanent switch of mitotic to stably postmitotic phentypes, fibroblasts displayed growth suppression and morphological changes of cell senescence;② increasing expression of SA-β-galactosidase;③ increasing expression of p16 protein;④ continuous up-regulation of mRNA of MMP-1 and MMP-3, while the protein of TIMP-1 was only slightly induced;⑤ high levels of the 4977bp deleted mtDNA accumulated in dermis and cultured fibroblasts, and large accumulation of a A→C base transversion of 414 position of 〓 of human mtDNA control region for replication of cultured fibroblasts also.

2.8-MOP/UVA作用下，体内外真皮成纤维细胞生物学特性出现具有光老化特征性的改变：①细胞由具有分裂活性的分裂表型转化为不具有分裂能力的分裂后表型，形态学出现细胞衰老的相应改变；②SA-β-Gal表达增加；③p16蛋白表达增加；④MMP-1、MMP-3 mRNA持续表达而TIMP蛋白表达仅被轻微诱导；⑤mtDNA 4977bp缺失大量累积，培养成纤维细胞mtDNA复制控制区〓片段414碱基迅速出现大量A→C的点突变。

Adding dividedly the solution of two drugs with different concentration into the heperplastic scar fibroblasts and keloid fibroblast which were in logarithmic growth phase. After 48 hours in vitro culture, hydroxyproline colorimetric test was conducted, then statistics comparison was carried out in accordance with hydroxyproline content which was calculated based on OD value.

向对数生长期的增生性瘢痕成纤维细胞及瘢痕疙瘩成纤维细胞中加入不同浓度的两种药液，培养48小时后进行羟脯氨酸比色试验，并将 OD 值依公式换算成羟脯氨酸含量，进行统计学比较。

The study was done to compare the induction efficiency of human embryonic stem cells to neuroepithelial progenitor cells in human embryonic fibroblasts,mouse embryonic fibroblasts and feeder free systems.

中文摘要：［摘要］比较人胚胎干细胞在人胚胎成纤维细胞、鼠胚胎成纤维细胞饲养层和无饲养层3个诱导体系下形成神经上皮祖细胞的能力。

Coculturing of the transfected fibroblasts with neonatal rat ventricular myocyte cultures resulted in a significant reduction (68%) in the spontaneous beating frequency of the cultures compared with baseline values and cocultures seeded with naive fibroblasts.

结果发现，与基线、与幼稚成纤维细胞共培养相比，转染的成纤维细胞和新生大鼠心室肌细胞共培养导致培养物自发搏动频率的下降(68%)。

Methods: Fibroblast cell were treated with -300,-500 and -1 000V PTFE electrets for 24, 48 and 72 h, respectively, and the influence of negative electrets on cell apoptosis was studied by means of flow cytometry and transmission electron microscope.

选用-300、-500和-1 000 V PTFE驻极体作用于成纤维细胞24、48和72 h，利用流式细胞仪和透射电子显微镜研究负极性驻极体对成纤维细胞凋亡的影响。

According to the clear water experiment, aeration performance of the new equipment is good with high total oxygen transfer coefficient and oxygen utilization ratio.

曝气设备的动力效率在叶轮转速为120rpm～150rpm时取得最大值，此时氧利用率和充氧能力也具有较高值。

The environmental stability of that world - including its crushing pressures and icy darkness - means that some of its most famous inhabitants have survived for eons as evolutionary throwbacks, their bodies undergoing little change.

稳定的海底环境─包括能把人压扁的压力和冰冷的黑暗─意谓海底某些最知名的栖居生物已以演化返祖的样态活了万世，形体几无变化。