转化生长因子-β1,2在不同发育阶段皮肤组织中的定位及表达量的变化

目的 :探讨转化生长因子 - β1,2 (TGF - β1和TGF - β2 )与α -平滑肌肌动蛋白 (α -ASMA)在胎儿和成人皮肤中的分布和表达变化规律。方法 :2 0例被测标本中包括不同胎龄的胎儿皮肤 15例和成人皮肤 5例。用免疫组化方法和病理技术确定这 3种蛋白在不同发育阶段皮肤中的定位和表达量。结果 :在妊娠早期的胎儿皮肤中 ,TGF - β1,2 和α-ASMA呈弱阳性表达 ,随着胎儿生长发育 ,这 3种蛋白的阳性细胞率逐渐增大 ;在妊娠晚期胎儿和成人皮肤中这 3种蛋白的表达量明显多于妊娠早期胎儿皮肤 (P <0 0 1)。TGF - β1,2 的阳性信号主要存在于表皮细胞 ,内皮细胞和部分成纤维细胞中 ,α -ASMA蛋白定位于肌成纤维细胞和汗腺上皮细胞内。结论 :TGF - β1和TGF - β2 可能在皮肤的发生、结构和功能的维持以及皮肤损伤后的修复中起重要作用。     

不同胎龄手掌侧皮肤皮嵴的分化特征

按常规石蜡包埋、连续切片,H—E染色和用3%KOH孵化除去表皮,然后用0.05%甲苯胺蓝染色两种方法,观察了胎龄10—22周胚胎手掌侧皮肤皮嵴分化、发育的形态学特征。(1) 12～14周。腺褶和乳头原基开始形成.(2) 14～16周,腺褶分化为汗腺原基,表皮开始角化。(3) 16～18周,沟褶发生,汗腺开始分化。(4)18～22周,真皮嵴明显,表皮嵴渐外显。     

不同胎龄的胎儿和少儿皮肤中bax,bcl-2和p53基因表达的变化

目的：探讨凋亡相关基因bax, bcl-2和p53在不同胎龄的胎儿皮肤和少儿皮肤组织中表达的变化特征及其可能的生物学意义。方法： 运用末端脱氧核糖转移酶介导的生物素化脱氧尿嘧啶缺口标记技术(TUNEL)检测18例不同胎龄(13-32周)的胎儿皮肤和6例少儿皮肤组织中细胞凋亡的变化后，提取这些皮肤组织中的总RNA，分离mRNA，用RT-PCR方法检测bax, bcl-2和p53基因在不同组织中的表达变化特征。结果： 随着胎儿的生长发育，皮肤组织中的细胞凋亡率逐渐增加。在早期妊娠胎儿的皮肤中，bcl-2基因表达水平较高，随着胎龄的增加，bcl-2基因的转录本含量逐渐降低，在少儿的皮肤组织中，这种基因的表达量明显低于早期妊娠胎儿皮肤（P<0.01）。与bcl-2基因不同，在早期妊娠胎儿皮肤组织中，p53基因表达水平较低，而在晚期妊娠胎儿和少儿的皮肤内，该基因表达较强，而bax基因在不同发育时期的胎儿和少儿皮肤组织中表达差异不显著（P>0.05）。结论： 晚期妊娠胎儿和少儿皮肤组织中细胞增殖减缓，细胞趋向分化或凋亡的增加可能与p53基因表达增强，bcl-2表达降低相关；而p53表达降低，bcl-2表达升高可能是早期妊娠胎儿皮肤中细胞凋亡较少的机制之一。     

胎儿皮肤中CK20的表达及与Merkel细胞分布的相关性研究

目的研究不同发育时期胎儿皮肤表皮基底层中细胞角蛋白20（CK20）的分布、数量,以揭示Merkel细胞分布的规律。方法CK20是Merkel细胞特异而敏感的标记物,分别取42例不同胎龄（16—30周）胎儿掌、跖、胸、背部的全层皮肤,采用免疫组化SP法观察CK20。结果掌、跖部位CK20阳性细胞的数量显著高于胸、背部;胚胎16～20和21—25周显著高于26—30周。结论（1）表皮Merkel细胞的数量存在解剖部位的差异。（2）胚胎发育中期Merkel细胞的数量显著多于胚胎发育的晚期。（3）Merkel细胞在表皮突内的高表达可能与表皮突结构的形成有关。     

人胎肝大小与胎龄的关系

目前,对胎儿肝的形态结构和组织发生报道较多[1],但对于胎肝小同发育阶段肝的左右最大径和胎龄之间关系的观察还未见报道.为了进一步探讨胎肝大小与胎龄的关系,我们对不同胎龄的人胎肝外形进行观察和测量.计算了相关系数,旨在为影像学动态观察胎儿的发育[5]以及胎儿外科提供参考.     

利用载有角质细胞生长因子微囊的组织工程皮肤修复裸鼠皮肤缺损

背景：组织工程皮肤作为一项新兴技术拥有良好的应用前景。有研究表明,角质细胞生长因子可以促进表皮细胞增殖。 目的：观察荷载角质细胞生长因子纳米微囊的新型组织工程皮肤修复裸鼠皮肤缺损的效果和特点。 方法：构建荷载角质细胞生长因子的脱细胞真皮基质复合物;将人表皮干细胞群和成纤维细胞分离、培养,并且进行鉴定;将表皮干细胞群接种于复合物之上,观察其生长状况;将荷载角质细胞生长因子纳米微囊的组织工程皮肤移植于裸鼠皮肤缺损处,将无角质细胞生长因子纳米微囊的组织工程皮肤作为空白组,将其自体皮肤移植修复缺损组作为对照组,于移植后2,4,6周时观察皮片挛缩及组织学愈合情况,并应用抗人角蛋白10及β1-整合素免疫荧光检测修复区表皮和真皮层细胞来源、分化及生长情况。 结果与结论：表皮干细胞在复合物表面生长良好,黏贴紧密,可见有连接成片趋势的小圆形的表皮干细胞及多角形的终末表皮细胞,部分形成克隆团块。移植后第2,4,6周,荷载角质细胞生长因子纳米微囊组织工程皮肤修复裸鼠皮肤缺损的结果均优于空白组及对照组,移植的皮肤边缘与邻近皮肤完全融合,但存在一定程度的挛缩。修复区组织工程皮肤的表皮细胞分层良好并能产生角质层,同时,移植后8,10周,组织工程皮肤切片免疫荧光染色可以鉴别出少量β1-整合素阳性细胞,均为表皮干细胞或短暂扩充细胞。结果证实,荷载角质细胞生长因子纳米胶囊的新型组织工程皮肤修复裸鼠皮肤缺损的效果较好,优于普通组织工程皮肤及自体全厚皮片移植修复。 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

ES细胞源性表皮干细胞与类真皮构建组织工程皮肤的体内分化

目的以ES细胞源性表皮干细胞为种子细胞与类真皮构建组织工程皮肤,探讨其在体内的分化。方法胎鼠皮肤成纤维细胞和大鼠骨髓基质干细胞(BMSCs),分别与复合凝胶-明胶海绵构建类真皮(类真皮Ⅰ、Ⅱ),植入小鼠全层皮肤缺损创面,以生物膜为载体,把羊膜诱导后带有核标记的表皮干细胞覆盖在类真皮上,术后1～8周连续取材,苏木精-伊红染色,β1整合素、CK15、CK19、CK10和CEA免疫荧光双标和免疫组化观察。结果两组组织工程皮肤植入皮肤缺损3～4周后,创面完全长合,较厚新生皮完全覆盖创面,基底层细胞增生,形成短的细胞柱突向真皮层。新生表皮中可见核标记的细胞呈β1整合素、CK15、CK19阳性,真皮中的管腔样结构呈核荧光和CEA免疫组化双标阳性,4～8周新生表皮基底层细胞呈CK19、CK10阳性,新生表皮下可见毛囊样、皮脂腺样结构。结论ES细胞源性表皮干细胞为种子细胞与类真皮构建的两种组织工程皮肤在体内具有修复缺损皮肤及分化为表皮及毛囊样、皮脂腺样和汗腺样等皮肤附属结构的潜能。     

组织工程皮肤的现状和展望

皮肤损伤及修复是临床工作者面临的重大难题。自体皮肤移植因供体不足而受限,而异体移植则面临免疫排斥反应。利用组织工程技术制作组织工程皮肤因而成为有效的解决途径。随着种子细胞和支架材料等的不断完善,组织工程皮肤的基础研究将更加丰富,临床应用的前景更加广阔。本文从组织工程皮肤的结构、分类、应用、产品等方面对该领域的国内外最新研究现状进行综述,分析了组织工程皮肤目前存在的问题,并提出未来发展方向即再生出含有毛囊、汗腺等附属器官的完整有功能的人类皮肤,对今后组织工程皮肤的研究和应用具有一定的参考价值。     

灵芝多糖抗氧化、抗皮肤衰老

背景：研究结果表明,灵芝多糖具有抗病毒、抗肿瘤、提高免疫力、抗氧自由基、抗衰老等生物活性。 目的：观察灵芝多糖对D-半乳糖致衰老小鼠皮肤组织抗氧化能力的生物学效应和延缓皮肤衰老作用。 方法：将44只2月龄昆明种小鼠随机分为4组,即正常组、衰老模型组、维生素E组、灵芝多糖组,后3组颈背皮下注射D-半乳糖建立小鼠衰老模型,同时灌胃给予相应药物或生理盐水,42 d后取小鼠背部皮肤作病理切片,观察皮肤组织形态变化,检测表皮、真皮厚度,测定超氧化物歧化酶含量及CuZn-SODmRNA在皮肤中的表达水平。 结果与结论：维生素E组和灵芝多糖组小鼠表皮、真皮厚度均较衰老模型增加。灵芝多糖组小鼠皮肤超氧化物歧化酶活力明显高于其他组。灵芝多糖组小鼠皮肤Ct值降低显著低于其他组。说明灵芝多糖能增加表皮和真皮厚度,改善皮肤组织结构;提高皮肤组织超氧化物歧化酶水平及CuZn-SOD mRNA的表达。     

转化生长因子-β1,2在不同发育阶段皮肤组织中的定位及表达量的变化

目的:探讨转化生长因子-β_{1, 2}(TGF-β₁和TGF-β₂)与α-平滑肌肌动蛋白(α-ASMA)在胎儿和成人皮肤中的分布和表达变化规律。方法:20例被测标本中包括不同胎龄的胎儿皮肤15例和成人皮肤5例。用免疫组化方法和病理技术确定这3种蛋白在不同发育阶段皮肤中的定位和表达量。结果:在妊娠早期的胎儿皮肤中, TGF-β_{1, 2}和α-ASMA呈弱阳性表达, 随着胎儿生长发育, 这3种蛋白的阳性细胞率逐渐增大;在妊娠晚期胎儿和成人皮肤中这3种蛋白的表达量明显多于妊娠早期胎儿皮肤(P＜0.01)。TGF-β_{1, 2}的阳性信号主要存在于表皮细胞, 内皮细胞和部分成纤维细胞中, α-ASMA蛋白定位于肌成纤维细胞和汗腺上皮细胞内。结论:TGF-β₁和TGF-β₂可能在皮肤的发生、结构和功能的维持以及皮肤损伤后的修复中起重要作用。     

功能性组织工程皮肤附属物再生对排汗和新陈代谢的意义

背景：排汗可将一些溶解于汗液的代谢产物带出体外,在排泄中起辅助作用。现代医学目前还无法恢复严重创伤或烧伤患者的汗腺,如何解决皮肤汗腺的再生及如何培养具有皮肤附属物的“功能性组织工程皮肤”问题已成为当今医学研究中的热点。 目的：探讨机体的新陈代谢、汗液的代谢方式,分析组织工程皮肤功能性附属物汗腺再生研究的现状。 方法：收集组织工程皮肤功能性附属物再生,特别是排汗的相关研究,分析机体的新陈代谢与排泄方式,汗液代谢的物质和方式,以及目前组织工程皮肤附属物汗腺再生研究的现状。 结果与结论：汗液作为排泄系统中重要的一员在维持机体内部的动态平衡,防止代谢产物对身体的损害方面具有不可忽视的地位。理想的功能性组织工程皮肤应具有分泌汗液、排泄废物、调节体温和维持皮肤动态平衡的作用,在大面积烧伤后皮肤创伤修复和重建过程中起着至关重要的作用。已有研究将干细胞与汗腺细胞进行共培养,然后移植到瘢痕创面上进行重建汗腺组织,取得了一定的成功。     

Immunolocalization of keratin proteins in sweat gland tumours by the use of monoclonal antibody

A total of 34 cases (eccrine poroma: 2, cylindroma: 2, eccrine spiradenoma: 4, syringocystadenoma papilliferum: 1, hydroadenoma papilliferum: 1, clear cell hydroadenoma: 7, mixed tumour: 16) of sweat gland tumours of the skin were described in terms of immunohistochemical distribution of keratins using polyclonal anti-keratin antiserum (TK, detecting 41-56 KDa keratins) and monoclonal antibodies (KL1, 55-57 KDa; PKK1, 40, 45, 52.5 KDa). Keratin expression in eccrine poroma, spiradenoma and syringocystadenoma was similar to that in the ductal segment of normal sweat glands. Cylindroma showed usually slight staining for kertins. Tumour cells of hydroadenomas showed not so prominent staining for any of the keratins; however, histologically, tumour cells indicated marked variation, and the degree of keratin proteins also was different among these histological variants. Mixed tumours of the skin were strongly decorated with anti-keratin antibodies along the luminal surface cells of typical structures, while no staining occurred in outer side cells. Luminal tumour cells may be differentiated from secretory coil cells, whereas outer side cells may have a myoepithelial origin, as outer layer cells found in pleomorphic adenoma of salivary glands.     

Prenatal development of human major salivary glands and immunohistochemical detection of keratins using monoclonal antibodies.

The major salivary glands were examined from 69 human fetuses ranging from 10 to 40 weeks of gestation. Prenatal growth curves of developing salivary glands could be established by histological scoring, and development was divided into the early developmental stage (EDS) from 10 to 18 weeks, early intermediate developmental stage (EIDS) from 19 to 24 weeks, late intermediate developmental stage (LIDS) from 15 to 32 weeks, late developmental stage (LDS) from 33 to 40 weeks. Characteristic morphogenesis and cytodifferentiation occurred in glandular duct cells during the period of EIDS and LIDS. In the LDS, acini and ducts of the salivary glands histologically developed into a mature state similar to adult glands. Immunohistochemical staining with monoclonal antibodies (MoAbs) PKK1, KL1, K8.12, K8.13, K4.62, RPN 1160, 1162, 1163, 1164, and 1165 was performed. During the fetal period, keratin expression as revealed by MoAbs PKK1, KL1, K8.12 was well established, and the staining pattern for each of these antibodies was comparable. Other antibodies showed rare or negative staining except K8.13 which had a diffuse, non-specific staining pattern. Accordingly, the proliferation and cytodifferentiation of fetal stage keratin staining in ductal cells as revealed by MoAbs PKK1, KL1, and K8.12 showed a heterogenic distribution in both luminal and basal cells. It is a characteristic finding that the cytodifferentiation of ductal luminal cells precedes ductal basal cells. Ductal basal cells stained with MoAb K8.12 and show heterogeneity of keratin distribution continuously until the full term of gestation. The keratin staining of oral epithelium was also examined to compare with distribution of salivary gland ductal cells and oral epithelial cells. In the present study, the developmental sequence of salivary gland cells and the immunohistochemical properties of keratin proteins in these cells were described in relation to the histogenesis of salivary gland tumours.     

Matrigel‐Induced Tubular Morphogenesis of Human Eccrine Sweat Gland Epithelial Cells

Human eccrine sweat glands are tubule‐structured glands of the skin that are vital in thermoregulation, secretion, and excretion of water and electrolytes. A study of tubular morphogenesis in vitro would facilitate the development of a tissue engineering model for eccrine sweat glands and other tubule‐structured glands. Matrigel, a basement membrane matrix, has been shown to promote differentiation and morphogenesis of many different cell types, including tubular cells. This study investigated the growth, differentiation, and tubular morphogenesis of human eccrine sweat gland epithelial cells cultured in Matrigel. Human eccrine gland epithelial cells were isolated and cultured in vitro. The cell growth in Matrigel was evidenced by the formation of cell clusters, which were observed under an inverted microscope. The internal structure of the cell clusters was further investigated by hematoxylin–eosin (HE) staining and confocal laser scanning microscopy (CLSM) of propidium iodide‐stained nuclei. The results demonstrated that although on a plastic surface or in a collagen gel the cells could not form tubular structures, they formed tubular structures when cultured in Matrigel. Consequently, we conclude that Matrigel can promote tubular morphogenesis of human eccrine sweat gland epithelial cells. Anat Rec, 2011. © 2011 Wiley‐Liss, Inc.     

胎儿下颌下腺内瘦素和瘦素受体的表达及定位

目的：观察胎儿下颌下腺内瘦素和瘦素受体的表达、分布及发育规律。方法：HE染色法和免疫组织化学SABC法。结果：胎儿下颌下腺内纹状管和小叶问导管上皮细胞呈瘦素及瘦素受体免疫阳性反应,免疫反应产物分布于导管上皮细胞胞质内,细胞核星阴性反应。在胚胎发育的不同阶段,免疫反应强度亦不等。结论：瘦素和瘦素受体表达于胎儿下颌下腺内,可能参与调节胎儿下颌下腺和胃肠的发育及功能活动。     

小鼠下颌下腺发育中转化生长因子β受体的表达

背景：小鼠的下颌下腺是研究唾液腺的发育的良好模型,转化生长因子β是器官发育和疾病中重要的生长因子,但是在下颌下腺中转化生长因子β受体的表达以及作用机制至今并不明确。 目的：观察胚胎小鼠下颌下腺发育过程中转化生长因子βⅠ型受体和Ⅱ型受体以及p-ERK1/2的表达,揭示转化生长因子β在小鼠涎腺发育中的作用。 方法：取C57BL/6J小鼠胚胎期第14.5天的标本,使用转化生长因子βⅠ型受体和Ⅱ型受体以及p-ERK1/2抗体,对小鼠的下颌下腺进行免疫组化染色。取新生小鼠标本,大体观察下颌下腺,并且使用苏木精-伊红染色观察其形态。 结果与结论：①小鼠出生时,下颌下腺位于下颌骨下方;苏木精-伊红染色发现小鼠下颌下腺的腺泡、导管和闰管细胞也已经分化完成。②在胚胎期第14.5天,转化生长因子βⅠ型和Ⅱ型受体在腺泡上皮和导管上皮内高表达,而腺体上皮细胞外的间充质没有表达。③p-ERK1/2主要也是表达在下颌下腺的上皮细胞中,与转化生长因子βⅠ型受体和Ⅱ型受体在下颌下腺中的表达基本一致。说明在小鼠下颌下腺的发育过程中,转化生长因子β蛋白可能通过与上皮细胞表面的受体结合,激活ERK信号通路来调节涎腺腺泡和导管的发育。     

Hepatitis C virus replicates in sweat glands and is released into sweat in patients with chronic hepatitis C

Hepatitis C virus (HCV) replicates in salivary glands of chronic hepatitis C patients and is released into the saliva, suggesting that HCV may replicate in other exocrine glands. The presence of positive and negative HCV RNA strands was demonstrated by in situ hybridization, and of HCV core protein by immunohistochemistry, in sweat glands and keratinocytes in healthy skin biopsies from 15 patients with chronic hepatitis C and 10 anti-HCV negative patients with chronic liver disease. Positive and negative HCV RNA strands were detected in 9.6 +/- 5.2% and 4.2 +/- 3.8%, respectively, of the epithelial cells of eccrine sweat glands. Core protein was detected in 6.0 +/- 3.93% of these cells. HCV RNA resistant to RNase digestion (encapsidated HCV RNA) was detected in 10/10 sweat samples from HCV-infected patients. Positive and negative HCV RNA strands were detected in 6.7 +/- 2.97% and 3.0 +/- 3.08% of the keratinocytes, respectively. HCV core protein was found in 4.5 +/- 2.76% of these cells. No HCV RNA or HCV core protein was detected in the skin biopsies from the 10 anti-HCV negative patients. In conclusion, HCV replicates in eccrine sweat glands cells and keratinocytes in healthy skin and is released into the sweat.     

Neuroblastic nodules and giant epithelial cells in suprarenal gland in fetuses of different gestational age groups

Introduction: Microscopic neuroblastic nodules are found in the suprarenal gland of newborns and young infants. The purpose of this study was to determine the morphology and derivation of neuroblastic nodules and giant epithelial cells of human fetal suprarenal gland. Materials and methods: The study consisted of 30 spontaneously aborted human fetal specimens from 11th weeks to 28th weeks of gestational ages. The suprarenal glands were taken from fetal specimens for histological study. The staining was done by hematoxylin and eosin. The following points about the neuroblastic nodule and giant epithelial cells were noted: number, location, size, any specific change like cystic degeneration with increasing age in the cells, distribution pattern of neuroblasts and giant epithelial cells, and their relation to the capsule, cortex, and central vein. Results: The neuroblastic nodules and giant epithelial cells increased in number as the gestation advanced but after 20 weeks of gestation, their number started decreasing. At the gestational age 25–28 weeks, both the giant epithelial cells and neuroblastic cells were absent. Conclusion: Both giant epithelial cells and neuroblastic cells are an integral part of the normal development of the suprarenal gland and they disappear with the advancement of the gestational age. These cells are important because some authors consider them to be the precursor of malignancy in later life.     

A simple method of preparation of keratin filaments and production of a polyclonal broad-spectrum anti-cytokeratin antiserum for immunohistochemical application in fixed and unfixed epithelial tissues

Rabbits were immunized with 10 nm filaments of a mixture of cytokeratins which has been isolated from human heel callus material and reconstituted to filaments in vitro. The antisera to keratins (ASK) have been tested histologically at fixed and unfixed tissue samples by means of the indirect immunofluorescence and PAP technique. The ASK recognized specifically only the epithelial cells of skin, of the mucous membranes of mouth and digestive tract, of salivary glands, sweat gland and mammary gland, but did not react with hepatocytes or kidney cells. The following tumors, tested till now, reacted with the antikeratin antisera: epithelial and lymphoepithelial carcinomas of skin, mouth and digestive tract, carcinomas of salivary glands, mammary gland and thyroid gland, adamantinoma, basalioma of skin, and metastases from carcinomas.