用基因芯片研究不同皮肤组织创面愈合过程中同源异形框基因的差异表达

目的 探讨人胎儿及成人皮肤创面愈合过程中同源异形框基因的差异表达.方法 采用包含有14000个靶基因cDNA克隆的基因芯片技术,对正常成人和胎儿及其创面愈合过程中同源异形框基因族的表达进行对比观察.结果 正常成人和胎儿及其创面愈合过程中同源异形框基因族呈现差异表达,其中以PRX-2、HOXB13、HOXB6和HOXB7的差异表达为显著.结论 同源异形框基因的差异表达表明正常成人和胎儿皮肤的发育生物学状态存在差异,可能是胎儿和成人皮肤不同创面愈合方式的根本原因.     

胎儿和成人皮肤及其创面愈合过程中碱性成纤维细胞生长因子的表达及意义

目的　探讨胎儿和成人皮肤及其创面愈合过程中碱性成纤维细胞生长因子 (b FGF)的表达及其意义。方法　将孕龄 2 0～ 2 4周胎儿皮肤移植至 BAL B/ C裸鼠背部皮下 ,皮片成活后制造创面 ,建立胎儿无瘢痕愈合动物模型 ,定期获取相应标本。对临床所取正常成人皮肤及创面愈合皮肤标本 ,采用免疫组织化学染色方法 ,观察 b FGF的表达情况。　结果　正常胎儿皮肤及创伤后胎儿皮肤中均未见明显的 b FGF阳性表达。正常成人皮肤中血管周围可见阳性表达 ;创伤后成人皮肤也可见阳性表达 ,尤其成纤维细胞和血管内皮细胞创伤后表达明显增强。高倍镜视野随机观察计数b FGF阳性表达细胞数 ,正常胎儿皮肤为 2 .1± 0 .1,创伤后 12小时 ,1、3天和 1周胎儿皮肤分别为 2 .2± 0 .1、2 .1± 0 .3、2 .1± 0 .3和 2 .0± 0 .1;正常成人皮肤为 2 3.2± 4 .2 ,创伤后成人皮肤为 4 0 .5± 3.6 ,胎儿正常皮肤和创伤皮肤 b FGF表达与正常成人皮肤和创伤后皮肤 b FGF表达比较 ,差异有统计学意义 (P<0 .0 1)。　结论　 b FGF的阴性表达可能是胎儿皮肤无瘢痕愈合的重要原因之一。     

同源框基因NKX3.1在前列腺癌组织中的表达及意义

目的　探讨同源框基因NKX3.1在前列腺癌中的表达意义。　方法　采用半定量RT PCR检测前列腺癌、良性前列腺、非前列腺组织标本NKX3.1mRNA表达状况并对其扩增产物进行测序。　结果　 76例前列腺组织中 ,NKX3.1表达 75例 ,表达率 98.7%。 96例非前列腺组织标本中 ,除 2例睾丸组织、1例乳腺组织有NKX3.1表达外 ,肾、膀胱、肝、回肠、脂肪、皮肤组织无 1例表达。前列腺癌组织NKX3.1表达量明显增高 ,良性前列腺增生组织表达量明显减弱 (P <0 .0 1) ;晚期前列腺癌组织NKX3.1表达明显高于早期前列腺癌 (P <0 .0 5 ) ;低分化前列腺癌表达高于中高分化前列腺癌 (P <0 .0 5 ) ;激素依赖性前列腺癌表达高于激素非依赖性前列腺癌 (P <0 .0 1)。NKX3.1表达高低与前列腺体积和血清总PSA浓度无关 (P >0 .0 5 )。　结论　NKX3.1基因具有前列腺组织特异性 ,其表达状况与前列腺癌分期、分级相关。NKX3.1表达对于判断前列腺癌病理生物学特性和前列腺癌治疗有重要意义。     

Np63α在创面愈合过程中表达改变的研究

目的：探讨p63基因编码的蛋白亚型△Np63α在创面愈合表皮再生中的表达及其生物学意义。方法：①在3周龄小鼠背部用直径1．6cm的环钻压切制备出一个2．0cm~2全层皮肤缺损的圆形创面,并将切除的皮肤原位缝合以覆盖创面。②将18只昆明小鼠随机分成两组,实验组12只小鼠均在背部用环钻制备圆形创面；而对照组6只小鼠不致伤。分别于1d、2d、3d、7d、14d、21d在实验组创缘和对照组小鼠背部相同部位切取全层皮肤组织标本,用抗鼠多克隆抗体P40进行抗△Np63免疫组化染色。结果：△Np63α在正常皮肤中的特征性表达模式是强阳性染色限于生发层的角质形成细胞核。伤后1～2d,表皮开始迁移,与对照组正常皮肤相比,迁移表皮舌基底层细胞△Np63α表达下调。伤后3d,△Np63α有较强的表达出现在创缘和迁移的表皮舌中,阳性染色限于基底层和其上方1～2层的角质形成细胞,而创面尚未被上皮覆盖处则没有阳性细胞。伤后5～7d,△Np63a有更强的表达出现在创缘和迁移表皮舌的基底层角质形成细胞。伤后14～21d创面愈合,△Np63α在覆盖创伤区表皮的基底层和棘层的细胞中高表达,并且其表达在表皮中长期和持续的存在。结论：△Np63α可能通过调控表皮修复中的细胞增殖和细胞迁移而参与正常皮肤的创面愈合过程。     

胎儿及成人皮肤创面愈合过程中PDGF和EGF表达的对比性研究

目的 探讨人胎儿和成人皮肤及其创面愈合过程中PDGF和EGF的表达及意义。方法 利用已建立的人胎儿无瘢痕愈合动物模型，获取相应标本，结合临床所取成人皮肤标本，采用免疫组化染色方法，观察PDGF，EGF的表达情况。结果 ①正常胎儿皮肤中未见明显的PDGF阳性染色；创伤后12h、1d的胎儿表皮及真皮浅层可见PDGF的弱阳性表达；创伤后3d、1周的胎儿皮肤中PDGF的表达呈阴性；正常成人皮肤可在成纤维细胞，巨噬细胞及毛细血管见到阳性表达；创伤后表达加强；②正常胎儿皮肤表皮全层和毛囊，皮脂腺及汗腺细胞可见EGF的阳性表达；创伤后的胎儿皮肤中EGF的表达未见到明显变化；正常成人皮肤可见表皮基底层有中度阳性表达，毛囊，汗腺细胞也可见到轻度表达，创伤后表达有所减弱。结论 生长因子在胎儿和成人皮肤创面愈合过程中的差异表达可能是胎儿无瘢痕愈合的重要原因之一。     

△Np63α在创面愈合过程中表达改变的研究

目的:探讨p63基因编码的蛋白亚型△Np63α在创面愈合表皮再生中的表达及其生物学意义.方法:①在3周龄小鼠背部用直径1.6cm的环钻压切制备出一个2.0cm2全层皮肤缺损的圆形创面,并将切除的皮肤原位缝合以覆盖创面.②将18只昆明小鼠随机分成两组,实验组12只小鼠均在背部用环钻制备圆形创面;而对照组6只小鼠不致伤.分别于1d、2d、3d、7d、14d、21d在实验组创缘和对照组小鼠背部相同部位切取全层皮肤组织标本,用抗鼠多克隆抗体P40进行抗△Np63免疫组化染色.结果:△Np63α在正常皮肤中的特征性表达模式是强阳性染色限于生发层的角质形成细胞核.伤后1～2d,表皮开始迁移,与对照组正常皮肤相比,迁移表皮舌基底层细胞△Np63α表达下调.伤后3d,△Np63α有较强的表达出现在创缘和迁移的表皮舌中,阳性染色限于基底层和其上方1～2层的角质形成细胞,而创面尚未被上皮覆盖处则没有阳性细胞.伤后5～7d,△Np63α有更强的表达出现在创缘和迁移表皮舌的基底层角质形成细胞.伤后14～21d创面愈合,△Np63α在覆盖创伤区表皮的基底层和棘层的细胞中高表达,并且其表达在表皮中长期和持续的存在.结论:△Np63α可能通过调控表皮修复中的细胞增殖和细胞迁移而参与正常皮肤的创面愈合过程.     

创面愈合过程中EGF基因表达变化的研究

创面愈合是一个复杂的过程。文献中关于创面愈合过程中，上皮细胞生长因子（ｅｐｉｄｅｒｍａｌｇｒｏｗｔｈｆａｃｔｏｒ，ＥＧＦ）基因表达变化的研究较少，为此设计了本项研究。采用Ｗｉｓｔａｒ大鼠，在脊柱两侧制作１．５ｃｍ×１．５ｃｍ中厚断层皮肤缺损创面４个，于伤后第４，８，１２和１６天取材，用地高辛标记ＥＧＦｃＤＮＡ探针原位杂交方法，观察创面愈合过程中，ＥＧＦ基因活化表达的ｍＲＮＡ在组织中的分布变化。发现：在伤口愈合的全过程中，ＥＧＦ基因均有明显表达，以伤后第８天表达最强。提示：应用某种方法促进ＥＧＦ基因表达，可能会促进伤口愈合     

结直肠癌组织中同源异形盒基因B13与c-myc的表达及意义

目的:探讨同源异形盒基因B13(HOXB13)和c-myc在结直肠癌中表达的变化及意义。方法:用RT-PCR法和Western blot法检测54例结直肠癌患者癌组织及相应癌旁组织中HOXB13与c-myc的表达,并分析两者表达与结直肠癌的临床病理特征的关系。结果:结直肠癌组织中的HOXB13 m RNA和蛋白的表达明显低于癌旁正常结直肠组织,而c-myc m RNA和蛋白的表达明显高于癌旁正常结直肠组织(均P<0.05);两者的表达均与淋巴转移以及TNM分期有关(均P<0.05);结直肠组织中HOXB13蛋白与c-myc蛋白的表达呈负相关(r=-0.572,P<0.001)。结论:HOXB13的表达降低可能促进了原癌基因c-myc的表达,从而促进结直肠癌的发生、发展和转移。     

基因转染与创面愈合

基因治疗是最近几年兴起的一种新的治疗方法,应用基因工程技术促进创面愈合,为烧伤或慢性溃疡提供了一种全新的手段,共技术关键是将目的基因导入靶细胞并在起有效的表达,进而发挥其表达蛋白的生物学作用,本文就近年与创面愈合有关的基因转染,基因转染受体细胞的选择,转染功体特点,已转染的基因及其生物学作用进行综述。     

前列腺特异性同源框基因NKX3．1研究进展

NKX3．1最新近发现的前列腺特异性和雄激素调节基因，属于同源框基因家族，人类NKX3．1基因位于染色体8p21上，在人前列腺组织高度表达，为前列腺组织特异性，与前列腺组织发生和发育有关。是前列腺特异性肿瘤抑制基因，NKX3．1基因在前列腺组织表达状态呈现雄激素时间和浓度依赖性，并与前列腺癌发生发展以及前列腺癌的类型，分期和体积关系密切，有望成为诊断和治疗前列腺癌的有效工具。     

烧伤创面愈合的理论探索与临床实践

The basic and clinical research in wound healing have made great progress in China in the past 50 years. The method of " intermingle skin transplantation" which was first advocated by surgeons of Ruijin Hospital in 1966 greatly reduced the amount of autologous donor skin, thus making the coverage of an extensive burn wound possible. This method is al so known as " Chinese therapy". In 1986,doctors of Jishuitan Hospital reported successful coverage of an extensive burn wound with mieroautografts and allogeneic skin. The basic research of wound healing has been carried out since 1992,a series of studies showed the characteristics of biological behaviours of cells in concern, extracellular matrix and growth factor, the mechanism underlying progressive injury in deep second burn wound, the effect of " skin island" and the local immune tolerance induced by it (which are the key factors of intermingle transplantation).The induction of local immune tolerance has now become the re search hot subject of skin transplantation immunology. Stem cell research in the field of wound healing has been extensively car ried out. The theory of " dermal template defection" has been proposed as one of the mechanisms of scar formation. On the other hand, great progress has been achieved in the treatment of bums on the basis of clinical researches. Doctors of PLA 304 hospital found that excision of eschar on patients with extensive deep burn injury at early shock stage greatly decreased the occurrence of complications and mortality. Doctors of Ruijin Hospital reported that healing of deep second burn wound could be improved by tangential excision of burn eschar within 24 hours after burn injury. Doctors of Xiang ya Hospital reported patients suffering from deep bums of the hands got satisfied functional restoration when treated with tangential excision of eschar while degraded dermal tissue could be retained with transplantation of autoskin grafts.     

Stem cells in burn wound healing: A systematic review of the literature

IntroductionSevere burns are often associated with high morbidity and unsatisfactory functional and esthetic outcomes. Over the last two decades, stem cells have generated great hopes for the treatment of numerous conditions including burns. The aim of this systematic review is to evaluate the role of stem cell therapy as a means to promote burn wound healing.MethodsComprehensive searches in major databases were carried out in March 2017 for articles on stem cell therapy in burn wound healing. In total 2103 articles were identified and screened on the basis of pre-determined inclusion and exclusion criteria.ResultsFifteen experimental and two clinical studies were included in the review. The majority of studies reported significant improvement in macroscopic burn wound appearance as well as a trend toward improved microscopic appearance, after stem cell therapy. Other parameters evaluated, such as re-vascularization, collagen formation, level of pro- and anti-inflammatory mediators, apoptosis and cellular infiltrates, yielded heterogeneous results across studies.ConclusionStem cell therapy appears to exert a positive effect in burn wound healing. There is, therefore, justification for continued efforts to evaluate the use of stem cells as an adjunct to first-line therapies in burns.     

封闭负压引流技术对创面愈合过程中原癌基因表达的影响

目的 研究封闭负压引流技术(vacuum-assisted closure，VAC)对启动创面愈合过程和减少细胞凋亡的作用。方法 用免疫组化法检测猪急性皮肤缺损创面和人慢性创面原癌基因c-myc、c-jun和Bcl-2表达变化，计算阳性表达细胞数和标记指数，观察创面愈合过程。结果 ①VAC治疗组猪急性皮肤缺损创面清洁无明显渗出，第6天即有较多新生上皮和肉芽组织，25d完全愈合。对照组创面有较多渗出和血痂，第6天出现少量新生上皮和肉芽组织，30d愈合。伤后即刻c-myc、c-jun和Bcl-2表达量少，主要位于基底细胞的胞浆或胞核，伤后及VAC治疗后表达迅速显著增加，但表达至峰值后迅速下降。在伤后或VAC治疗后的12d内实验组表达始终高于对照组。②人慢性创面VAC治疗后分泌物明显减少，较快形成健康的肉芽组织。c-jun主要表达在表皮基底细胞、真皮成纤维细胞和炎性细胞的胞浆，VAC治疗后阳性细胞数和标记指数显著减少。c-myc和Bcl-2主要表达在基底细胞的胞浆，VAC治疗后表达量显著增多，标记指数显著增大。结论 VAC能快速启动猪急性皮肤创面和人慢性创面的愈合过程，减少修复细胞凋亡，使创面愈合加速。     

mRNA and protein expression of transcription factor c—fos in burned rats and their effects on wound healing

Objective:To explore the expression of mRNA and its protein in burned rats and their effects of burn wound healing.Methods:A partial-thickness burn of 30% total body surface area was created on the back of 40 Wistar rats.In situ hybridization and immunohistochemical methods were used to exaluate the location and the amount of the c-fos mRNA and its protein in normal skin and the burned skin,respectively,at 3h,6h,1d,3d,7d and 14d after burn.Results:Under a light microscope,both the expression of c-for mRNA and its protein could be found in the normal skin,but their induction levels were much higher in the burned skin.The level of for protein expression reached peak at 3h after burn while that of c-for mRNA reached peak at 6h after burn.Conclusions:The expression of c-fos can be induced by burns.And the peak level expression of c-for mRNA comes later than that of c-fos protein.It indicates that the action of fos protein is induced by post-translational modification of pre-existing fos molecules.     

金属蛋白酶组织抑制因子与皮肤创伤愈合

基质金属蛋白酶 (ｍａｔｒｉｘＭｅｔａｌｌｏｐｒｏｔｅｉｎａｓｅｓＭＭＰｓ)是参与细胞外基质 (ＥＣＭ)降解的主要蛋白酶 ,金属蛋白酶组织抑制因子 (ＴｉｓｓｕｅＩｎｈｉｂｉｔｏｒｏｆＭｅｔａｌｌｏｐｒｏｔｅｉｎａｓｅｓＴＩＭＰｓ)是一族内源性抑制因子 ,可以高度地、有选择地抑制ＭＭＰｓ或其前体而表现出许多生物学活性。ＴＩＭＰｓ和ＭＭＰｓ的平衡失控可导致创伤延迟愈合和其它许多疾病。现就有关ＴＩＭＰｓ的种类、结构和生物学作用及在皮肤创伤愈合中作用等研究综述如下。1　ＴＩＭＰｓ的产生和结构目前已知ＴＩＭＰｓ家族有…     

不同浓度法舒地尔促进皮肤创面愈合作用的研究

目的：观察法舒地尔（HA1077）促进创面愈合的效果,并探讨HA1077与创面愈合的量效关系,寻找出促进创面愈合的最佳剂量。方法：18只Wistar大鼠背部左右两侧致直径为2cm的圆形皮肤缺损。随机3只动物6个创面为一组,分为6组,分别给予10、20、40、80和160μmol/L盐酸法舒地尔和生理盐水（对照组）创面喷洒,每个创面0.5ml,隔日创面追加喷洒。实验3、7、10d计算伤口面积,实验10d取创面组织,观察组织学变化。结果：各组大鼠创面面积随伤后时间延长而逐渐缩小,20μmol/L组创面面积明显小于同时间点其他各组,除实验7d与80μmol/L组创面面积比较差异无显著性外,其余各时间点差异均有显著性（P〈0.05）。病理学变化显示,创伤后10d,应用20μmol/L法舒地尔组创面新生肉芽组织生长及新生表皮生长速度明显优于其他各组。结论：HA1077可促进皮肤缺损创面愈合,以20μmol/LHA1077效果更为明显。     

创伤愈合过程中酸性粘多糖的定位分布

观察胚胎兔、成兔皮肤在正常发育及创伤愈合过程中羧基、硫酸基粘多糖的定位分布。利用胎兔创伤模型,切取皮肤组织标本,用Ｓｐｉｃｅｒ的特殊染色方法切片观察。随胎兔不断成熟,表皮层中除基底层外羧基粘多糖逐渐减少,真皮层呈一致弥漫的分布,成兔表皮层主要为硫酸基粘多糖,真皮网状层的羧粘多糖多围绕胶原纤维分布,密度较深,两者羧基粘多糖染色最浓的部位位于基底层附近。创伤后,胎及成兔均表现出表皮基底层及真皮层羧基粘多糖合成增加,两者的差别不甚显著,而胎兔则在创伤后第５天完全愈合。硫酸基粘多糖的分布部位主要在表皮浅层和真皮层的毛囊细胞、血管及腺体上皮之中,创伤后反应不明显。羧基粘多糖是参与创伤愈合反应的主要细胞外基质,在创伤愈合及瘢痕形成领域具有十分重要的作用。     

Nerve growth factor and burn wound healing: Update of molecular interactions with skin cells

Burn wound healing is a very intricate and complex process that conventionally includes three interrelated and overlapping stages of hemostasis/inflammation, proliferation and remodeling. This review aims to explore the molecular interactions of NGF with the most prominent cell types in the skin and their respective secretory products during wound healing, particularly burn wound healing. Different types of cells such as, nerve cells, endothelial cells, mast cells, macrophages, neutrophils, keratinocytes and fibroblasts all come into play through a plethora of cytokines and growth factors including nerve growth factor (NGF). NGF is a pleiotropic molecule that exerts its effects on all the aforementioned cells using two types of receptors (TrkA and p75) and affects wound healing by decreasing healing time and improving the quality of the scar. Both receptors mediate cellular proliferation, survival and apoptosis through complex signaling molecules. During the inflammatory phase, macrophages and mast cells secrete ample cytokines and growth factors, including NGF, which participate in the inflammatory reaction and induction of other cells targeting a homeostatic state. The proliferative phase follows, and NGF promotes angiogenesis through VEGF and FGF expression in endothelial cells. NGF also stimulates keratinocyte proliferation and neurite extension through the TrkA-PI3K/Akt pathway. Other molecules such as TGF-β1, IL-1β and TNF-α increase NGF expression in fibroblasts through dynamic interactions with Smads and MAPK molecules. Stimulated fibroblasts induce new collagen production to form the granulation tissue. In the remodeling phase, NGF regulates fibroblasts and induces their differentiation into myofibroblasts ultimately leading to wound contracture. In addition, NGF stimulates melanocytes and enhances hair growth and pigmentation. Such data depict the mechanisms of action of NGF implicated in the various stages of the healing process and support its applicability as a new targeted therapeutic molecule effective in burn wound healing but with some limitations.     

更进一步提高深度烧伤创面修复质量

This article summarizes methods of repair of massive and deep wounds, elucidates how to improve wound healing quality and avoid scar deformity after deep hum. A part of denatured dermis (non-necrotic)in deep partial-thickness burn, "mixed degree" burn, even in full-thickness burn wounds before forming eschar can be preserved and covered with autolo-gous skin, thereby to avoid secondary damage to the structure of subcutaneous tissue and the junction of dermis-adipose, thus to result in good functions, appearance, and survival rate. After skin grafting, wound healing quality and appearance are im-proved, joint function and elasticity of skin are enhanced, the degree of scar contracture is relieved due to preservation of nor-mal adipose tissue after escharectomy. The study of composite artifical skin will be actively developed in the future. Tissue-en-gineering skin and stem cells can be successfully used in pa-tients with deep burns for starless healing with restoration of physiological functions in a short period.