颧骨缝牵张成骨过程中组织超微结构变化的研究

目的:观察天然骨缝牵张成骨过程中组织和细胞超微结构的变化。方法:对牵张成骨的缝区组织经过系列处理,进行超薄切片、透射电镜观察。结果:间充质细胞在牵引早期大量增生,并不断分化为成纤维细胞和成骨细胞,后2种细胞的超微结构显示出具有活跃的合成和分泌功能。1周标本成纤维细胞沿牵引力的长轴方向排列,胞核增大,在其周围包绕着发达并扩张的内质网系统;3周标本大量增生活跃的成骨细胞和成纤维细胞,成骨细胞核仁增大、粗面内质网扩张、核糖体丰富、线粒体增多、富含紧密排列的嵴;5、8周标本中骨细胞形成并发育成熟,骨基质逐渐矿化,清晰可见新形成的胶原纤维、哈氏管,以及骨基质矿化的过程。结论:三维牵张过程中,成纤维细胞、成骨细胞活跃,缝区形成新骨。     

大鼠肠缺血-再灌注损伤中FGFR2表达的规律

目的观察酸性成纤维细胞生长因子受体2（acid fibroblast growth factor receptor,FGFR2）在肠缺血-再灌注损伤中表达规律。方法以大鼠肠系膜上动脉（SMA）夹闭造成肠缺血-再灌注损伤模型,并将动物随机分为假手术组（C组）、生理盐水对照组（R组）、改构体aFGF治疗组（F组）。除假手术组外,其余各组动物均于缺血45min后于2、6、12、24h活杀,取小肠组织标本,免疫组化和RT—PCR检测FGFR的表达规律。结果在正常大鼠,FGFR分布在小肠绒毛上皮细胞的肠腔侧、侧壁和小肠隐窝朝向隐窝腔的-侧的细胞膜上。缺血和再灌注的初期,FGFR的表达未发生明显变化,随着再灌注时间的延长逐渐增强。FGF使FGFR的表达有明显的增强和提前表达。缺血和再灌注使FGFRmRNA的表达迅速增加,在再灌注后6h达到高峰。F组FGFRmRNA的表达较R组相应时间点显著增加（P〈0．05）。结论FGFR在肠缺血-再灌注损伤的修复中起积极作用。     

低氧诱导因子-1α在成纤维细胞中的表达及对毛囊的影响

目的：探讨外源性人低氧诱导因子（HIF）-1α基因在成纤维细胞中的表达及对体外培养人毛囊的影响。方法：通过脂质体将含有HIF—1α cDNA的真核表达载体pcDNA3．0瞬时转染成纤维细胞,应用反转录（RT）-PCR、免疫组化等方法检测HIF-1α在成纤维细胞中的表达。同时通过ELISA检测转染细胞上清液中血管内皮生长因子（VEGF）的表达情况。进一步将该上清加至体外培养的人毛囊和细胞中,显微镜下测量毛囊的平均生长长度,并观察毛囊的形态学变化。并通过四甲基偶氮唑蓝（MTT）法检测加入转染上清的细胞活性。结果：成功地将HIF—1α真核表达载体HIF—1α pcDNA3．0瞬时转染了成纤维细胞,用RT-PCR、免疫组化染色可检测出细胞中HIF—1α的表达,MTT检测转染后细胞活性增强,并且该上清液可以检测出VEGF的表达。该上清可以明显促进体外培养的人毛囊生长．延缓其进入退行期。结论：应用脂质体能够成功地将外源性人HIF—1α基因转染成纤维细胞,并进行有效表达,其表达的HIF—1α可增强细胞活性,且可诱导转染细胞上清液中VEGF的表达,在体外具有促进人毛囊生长的生物学活性。     

Influence of Indium (III) Chloride on Human Dermal Fibroblast Cell Adhesion on Tantalum/Silicon Oxide Nano-Composites

Cell adhesion is an essential biological function for division, migration, signaling and tissue development. While it has been demonstrated that this cell function can be modified by using nanometer-scale surface topographic structures, it remains unknown how contaminants such as indium (III) ion might influence this specific cell behavior. Herein, the influence of indium chloride on human dermal fibroblast (GM5565) adhesion characteristics was investigated, given the frequent contact of contaminants with skin. The morphology of the adherent cells and their mitochondrial reticulum was characterized on cell culture dishes and nanopatterned surfaces by using fluorescence confocal microscopy and scanning electron microscopy. Results showed a significant proportion of cells lost their ability to align preferentially along the line axes of the nanopattern upon exposure to 3.2 mM indium chloride, with cells aligned within 10° of the pattern line axes reduced by as much as ~70%. Concurrent with the cell adhesion behaviors, the mitochondria in cells exposed to indium chloride exhibit a punctate staining that contrasts with the normal network of elongated tubular geometry seen in control cells. Our results demonstrate that exposure to indium chloride has detrimental effects on the behavior of human fibroblasts and adversely impacts their mitochondrial morphology. This shows the importance of evaluating the biological impacts of indium compounds.     

肾衰保肾胶囊对肾间质纤维化大鼠肾脏MMP-1表达及活性的影响

目的:研究肾衰保肾胶囊对慢性肾间质纤维化大鼠基质金属蛋白酶1(MMP-1)表达及活性的影响。方法:将Wistar大鼠60只随机分为空白对照组、模型对照组、肾衰保肾胶囊低剂量组和肾衰保肾胶囊高剂量组,采用腺嘌呤灌胃复制慢性肾间质纤维化大鼠模型,各组分别给药8周,采用免疫组化法、RT-PCR及酶谱分析法检测肾组织中的I、Ⅲ型胶原、MMP-1表达及MMP-1活性。结果:肾衰保肾胶囊组大鼠肾组织中的I、Ⅲ型胶原表达降低,MMP-1表达及活性增强,与模型对照组比较有显著性差异(P<0.01)。结论:肾衰保肾胶囊可能是通过增强MMP-1表达及活性的途径来减少肾组织中I、Ⅲ型胶原的积聚,进而发挥减轻肾间质纤维化的作用。     

Quantitative Study of Basic Fibroblast Growth Factor in Vitreous with Proliferative Vitreoretinopathy

Objective: To quantitatively study basic fibroblast growth factor (bFGF) in the vitreous of proliferative vitreoretinopathy (PVR) in orderto understand the role of bFGF in the development of PVR.Method: High sensitive sandwich enzyme immunoassay technique(ELISA) was used to measure bFGF level in vitreous of normal eyes, the eyes of PVR-C or PVR-D grade, eyes of vitreous hemorrhage and the serum levels of bFGF in PVR-D patients. Results: The levels of bFGF in the vitreous were: median 5. 20ng/L, quartile 15. 47 ng/L in 20 normal eyes; median 3. 12ng/L, quartile 10. 48 ng/L in 35 PVR-C eyes; median 46. 56 ng/L, quartile 113. 96 ng/L in 26 PVR-D eyes; median 1. 40 ng/L, quartile 6. 25 ng/L in 25 vitreous hemorrhage eyes. The vitreous bFGF level in PVR-D group was significantly higher than that in the normal group, PVR-C group and vitreous hemorrhage group ( P < 0. 01). The mean of serum-bFGF level was 18. 33 ± 3. 39 ng/ L. The vitreous bFGF level of PVR-D group was significantly higher than serum-bFGF lev     

Role of chymase on growth of cultured canine Tenon's capsule fibroblasts and scarring in a canine conjunctival flap model

Chymase is a chymotrypsin-like serine protease contained in the secretory granules of mast cells. Recently, we reported that chymase activity and the number of chymase-positive mast cells in conjunctival tissues were significantly increased during the wound healing process in a hamster model of glaucoma surgery. However, it has been unclear the role of chymase on conjunctival scarring. In the present study, we evaluated the effect of dog chymase on cell proliferation of fibroblasts established from canine Tenon's capsule and the effect of a chymase inhibitor on scarring in a canine conjunctival flap model. After a fibroblast cell culture was established from canine Tenon's capsules, the fibroblasts were incubated in the presence of dog chymase (5-20 ng ml(-1)). Cell proliferation was evaluated by bromodeoxyuridine incorporation. In a canine conjunctival flap model, a sponge treated with a chymase inhibitor, Suc-Val-Pro-Phe(P)(OPh)(2), or placebo was placed in between the conjunctiva and sclera and the conjunctival incision was closed. One week after the surgery, adhesion degree was assessed, and chymase activities in the conjunctival lesion and in the areas of the conjunctiva and sclera were measured. In cultured canine Tenon's capsule fibroblasts, dog chymase significantly increased cell proliferation, and this chymase-dependent proliferation was completely suppressed by the chymase inhibitor. In the canine surgical model, chymase activity in placebo-treated eyes was significantly increased compared to control eyes, while it was significantly decreased by treatment with the chymase inhibitor. Scores for adhesion degree in the chymase inhibitor-treated eyes were significantly decreased in comparison with those in placebo-treated eyes. The conjunctival area in the chymase inhibitor-treated eyes was also suppressed to 52.6% compared with that in placebo-treated treated eyes. In conclusion, chymase stimulates proliferation of fibroblasts derived from canine Tenon's capsule and chymase may play an important role in scarring after glaucoma surgery.     

酸性成纤维细胞生长因子在宫颈癌中的表达及其信号传导途径

目的探讨酸性成纤维细胞生长因子(aFGF)在宫颈癌发生发展中的作用及其信号传导机制.方法应用逆转录-聚合酶链反应技术(RT-PCR)对36例宫颈癌组织aFGF 及其受体FGFR1的表达进行了分析;并以不同浓度的aFGF 和酪氨酸蛋白激酶(TPK)抑制剂genistein诱导宫颈癌细胞株HeLa细胞,[γ-32P]ATP掺入外源性底物的方法,液体闪烁测定蛋白激酶C(PKC)及细胞外信号调节激酶(ERK)的活性.结果 aFGF mRNA 和FGFR1 mRNA在宫颈癌组织中的半定量检测结果分别为(1.233±0.064)和(1.168±0.103),与正常宫颈组织相比,差异有显著性(P<0.05),且在Ⅲ～Ⅳ期宫颈癌中的表达水平明显高于Ⅰ～Ⅱ期(P<0.05);随着aFGF浓度的增加,HeLa细胞PKC及ERK 活性随之升高,与aFGF浓度呈剂量依赖效应;genistein抑制细胞内PKC及ERK 活性,与genistein 浓度亦呈剂量依赖效应.结论提示aFGF 与宫颈癌的发生、发展、浸润呈正相关,其受体具有TPK活性,TPK激活后可进一步激活PKC和ERK,进一步证明PKC及ERK确是TPK的下游信号分子.     

中药散血明目片抑制兔视网膜静脉阻塞后视网膜VEGF和bFGF的表达

目的:探讨活血通脉利水明目作用的中药散血明目片防治视网膜静脉阻塞(retinal vein occlusion,RVO)并发新生血管的作用。方法:将家兔48只96眼随机分为4组,每组12只24眼。A健康空白组;B模型组;C血栓通组;D散血明目片组。采用激光光凝法建立兔RVO模型,分别于术后3,7,14,21d用空气栓塞法处死动物,即刻摘取眼球石蜡包埋切片。免疫组化法观察兔视网膜组织中血管内皮生长因子(VEGF)和碱性成纤维细胞生长因子(bFGF)的表达。结果:成功建立了RVO模型。D组与B组比较,视网膜组织中VEGF含量(21d时分别为0.442±0.034和0.583±0.056)和bFGF含量(21d时分别为0.419±0.040和0.514±0.056)明显减少,差异有统计学意义(P<0.05),与C组比较差异无统计学意义。结论:活血通脉利水明目之散血明目片能抑制视网膜面VEGF和bFGF的高表达。     

Astrocytes in the damaged brain: Molecular and cellular insights into their reactive response and healing potential

Long considered merely a trophic and mechanical support to neurons, astrocytes have progressively taken the center stage as their ability to react to acute and chronic neurodegenerative situations became increasingly clear. Reactive astrogliosis starts when trigger molecules produced at the injury site drive astrocytes to leave their quiescent state and become activated. Distinctive morphological and biochemical features characterize this process (cell hypertrophy, upregulation of intermediate filaments, and increased cell proliferation). Moreover, reactive astrocytes migrate towards the injured area to constitute the glial scar, and release factors mediating the tissue inflammatory response and remodeling after lesion. A novel view of astrogliosis derives from the finding that subsets of reactive astrocytes can recapitulate stem cell/progenitor features after damage, fostering the concept of astroglia as a promising target for reparative therapies. But which biochemical/signaling pathways modulate astrogliosis with respect to both the time after injury and the type of damage? Are reactive astrocytes overall beneficial or detrimental for neuroprotection and tissue regeneration? This debate has been animating this research field for several years now, and an integrated view on the results obtained and the possible future perspectives is needed. With this Commentary article we have attempted to answer the above-mentioned questions by reviewing the current knowledge on the molecular mechanisms controlling and sustaining the reaction of astroglia to injury and its stem cell-like properties. Moreover, the cellular/molecular mechanisms supporting the detrimental or beneficial features of astrogliosis have been scrutinized to gain insights on possible pharmacological approaches to enhance astrocyte neuroprotective activities.