Adipose tissues differentiated by adipose-derived stem cells harvested from transgenic mice

Objective: To induce adipocyte differentiation in vitro by adipose-derived stromal cells (ASCs) harvested from transgenic mice with green fluorescent protein (GFP) and assess the possibility of constructing adipose tissues via attachment of ASCs to typeⅡcollagen scaffolds. Methods: Inguinal fat pads from GFP transgenic mice were digested by enzymes for isolation of ASCs (primary culture). After expansion to three passages of ASCs, the cells were incubated in an adipogenic medium for two weeks, and the adipocyte differentiation by ASCs in vitro was assessed by morphological observation and Oil Red O staining. Then they were attached to collagen scaffolds and co-cultured for 12 hours, followed by hypodermic implantation to the dorsal skin of nude mice for 2 months. The newly-formed tissues were detected by HE staining. Results: The cultured primary stem cells were fibroblast-like and showed active proliferation. After being incubated in an adipocyte differentiation medium, the lipid droplets in the cytoplasm accumulated gradually and finally developed into mature adipocytes, which showed positive in Oil Red O staining. A 0. 5-cm3 new tissue clot was found under the dorsal skin of the nude mice and it was confirmed as mature adipose tissues by fluorescent observation and HE staining. Conclusions: ASCs can successfully differentiate adipose tissues into mature adipocytes, which exhibit an adipocyte-like morphology and express as intracytoplasmic lipid droplets. It is an efficient model of adipose tissues engineered with ASCs and type I collagen scaffolds.     

类固醇类激素联合携带Fas基因重组腺病毒治疗瘢痕疙瘩的体内实验研究

目的应用成功制备的携带人Fas基因的两种重组腺病毒,联合类固醇激素进行瘢痕疙瘩的动物实验研究,判断携带人Fas基因的重组腺病毒与类固醇激素联合治疗瘢痕疙瘩的效果。方法构建瘢痕疙瘩裸鼠模型。使用Ad-Fas(B),Ad-Fas(T)两种构建成功的腺病毒注射及其它辅助治疗手段,实施针对植入裸鼠皮下的瘢痕疙瘩组织的体内治疗方案。通过大体观察,常规病理及电镜观察检测瘢痕疙瘩组织块的变化。结果①单纯使用腺病毒注射后的瘢痕疙瘩组织块体积仅轻度缩小。②前期注射Ad-Fas(B)或Ad-Fas(T)后,使用类固醇激素作为后续治疗因素,其瘢痕疙瘩组织块均明显缩小。③使用腺病毒治疗后,能有效地减少曲安缩松的使用,从而减轻类固醇类激素治疗的副作用。结论①裸鼠为免疫缺陷动物,所以该结果并不能否认病毒的直接治疗作用,在免疫性动物体内直接注射腺病毒的治疗效果尚无结论。②重组腺病毒Ad-Fas(B)及Ad-Fas(T)的瘢痕疙瘩基因治疗的动物实验为瘢痕疙瘩的治疗展示了一条全新的途径。     

瘢痕瘤的病因研究进展

瘢痕瘤是目前整形外科面临的重大难题,因此其病因研究也就成了目前整形外科研究的热点。本文介绍瘢痕瘤患者的遗传背景、动物模型的建立、成纤维细胞生物活性、胶原代谢异常、以及细胞因子的调控等瘢痕瘤病因研究领域内的研究进展。     

Differences between the gap junctional communication function of fibroblasts derived from pathological scars and normal skin

Keloid is a fibrotic disease Of the hUInan dends thattakes the form of benign fibIOus tumors resulting from anabnolmal response to wound. Unlike hyperplastic scarewith which keloid is freqUently collfused, keloid lesionusually extends beyond the bountyyes of the originalwound by invasive growth without rendssion with time,which is diffictilt to cormct s~cally and does not proVOke scar .on~tionll'2].It is noted that there ~ vallous patterns of intereellular interachon, as junchonal interoell…     

肿瘤标记物和CT联检对肺癌的诊断价值

目的:探讨多种肿瘤标记物检测联合CT扫描在肺癌诊断中的价值。方法:90例肺癌和20例肺良性病变患者与20名正常人,检测其血清NSE、CEA和CYFRA21-1三项肿瘤标记物含量,同时行肺部CT扫描检查。结果:肺癌组血清三项肿瘤标记物含量均高于正常对照组和肺良性病变组,P=0·000;NSE在小细胞肺癌中高表达(36·21±19·01)μg/L,CEA在肺腺癌中高表达(50·09±25·36)μg/L,CYFRA21-1在肺鳞癌中高表达(17·72±12·05)μg/L,三项肿瘤标记物的血清浓度与肺癌病理类型有关,P=0·001;肿瘤标记物和CT扫查对肺癌分型诊断总体符合率分别为79·2%和62·5%,联合检测的总体符合率为87·8%,联合检测优于单项检测,P=0·039;三项肿瘤标记物的含量在肺癌各期间差异无统计学意义,P>0·05。结论:多种肿瘤标记物联检,可提高肺癌分型的诊断符合率,结合CT分期有助于选择针对性的治疗方案。肿瘤防治杂志,2005,12(19):1480-1482     

缺氧对成熟脂肪细胞去分化影响的实验研究

目的 研究缺氧环境下成熟脂肪细胞体外培养的形态学变化,探讨缺氧环境对成熟脂肪细胞去分化的影响.方法 从成人吸脂术后抽吸物提取脂肪组织来源的干细胞,诱导为成熟脂肪细胞,在缺血缺氧的环境下培养.观察各时间段细胞的形态变化.结果 由脂肪组织来源的干细胞诱导而成的成熟脂肪细胞,在缺氧环境培养数日后,细胞从圆形变为可持续分裂的成纤维细胞状细胞.结论 成熟的脂肪细胞在一定条件下可以去分化为成纤维细胞状细胞.     

纤维蛋白胶促进自体脂肪移植存活的实验研究

[摘要]目的:探讨纤维蛋白胶是否能有效促进自体脂肪游离移植的存活率。方法：①选取清洁级新西兰大白兔10只,切取腹股沟区皮下脂肪,制成颗粒状。②以纤维蛋白胶结合自体脂肪颗粒（实验组）及生理盐水混合的自体脂肪颗粒（对照组）分别注射于兔腰背筋膜区对称部位的皮下间隙。③术后6个月取出脂肪移植物,比较两组移植物湿重,并切片HE染色比较微观形态差异。结果：①移植物湿重测量,实验组存活率为（64.9±12.2）%,远大于对照组,（29.2±9.3）%,差异有统计学意义（P<0.01）。②组织学切片HE染色显示实验组存留了更多的正常脂肪组织,实验组血管密度为（20.1±4.4）个/HF,大于对照组,（12.2±3.4）个/HF,差异有统计学意义（P<0.01）。结论：纤维蛋白胶基于其粘附性及促进新生血管形成的特性,可提高自体游离脂肪移植物的存活率。     

ARNT调控低氧通路促进小鼠骨骼肌再生的实验研究

目的　探索芳香烃受体核转运子（aryl hydrocarbon receptor nuclear translocator,ARNT）对骨骼肌再生的调控作用,为提高老龄人口骨骼肌再生能力提供方向。 方法　构建小鼠骨骼肌冰冻损伤模型,观察幼龄鼠与老龄鼠、骨骼肌特异性ARNT基因敲除小鼠与野生鼠、加入低氧通路激活剂（ML228）与DMSO老龄鼠间骨骼肌再生能力差别;分析ARNT、低氧通路、肌再生因子表达含量及小鼠下肢血流差异。 结果　衰老导致骨骼肌再生能力减弱;敲除ARNT基因后,小鼠骨骼肌的再生能力显著下降,低氧通路因子及相关基因表达下调;ML228可使受损的骨骼肌再生能力得到恢复。 结论　衰老引起的ARNT含量下降抑制低氧通路是导致老龄骨骼肌再生能力降低的主要原因。低氧通路激活剂可改善受损骨骼肌的再生能力,有望成为药物重塑衰老骨骼肌再生能力的新靶点。     

瘢痕疙瘩Fas基因(外显子6～9)突变的检测

瘢痕疙瘩是损伤愈合过程的结果 ,它们以超过原损伤范围的大量瘢痕组织形成为特征 ,而导致这种过度增殖的原因至今未明。近年来其它领域的研究证明 :Ｆａｓ系统对控制细胞增生与凋亡起重要作用 ,Ｆａｓ基因的体细胞突变与白血病、多发性骨髓瘤等疾病的发生、发展有着密切的关系。瘢痕疙瘩属良性肿瘤 ,是否也存在Ｆａｓ基因突变国内外尚未见报道。我们先前的实验研究提示了这种突变存在的可能性。本研究目的是检测瘢痕疙瘩成纤维细胞Ｆａｓ基因外显子 69是否存在突变及其突变位点。方法 :取 1 5例瘢痕疙瘩成纤维细胞进行ＰＣＲ ＳＳＣＰ检测 ,…