Subacute cutaneous lupus erythematosus leukoderma treated with melanocyte‐keratinocyte transplantation procedure

Depigmented lesions may occur as postinflammatory sequelae of subacute cutaneous lupus erythematosus (SCLE), leading to great psychosocial impact. A 53‐year‐old male patient presented with post‐SCLE depigmented facial lesions after five years of disease stability. We proposed surgical treatment with melanocyte‐keratinocyte transplantation procedure (MKTP), and after five months the patient achieved 90% repigmentation, without Koebner phenomenon (KP). In theory, KP is a possible complication of MKTP procedure since the preparation of the receptor area involves the use of dermabrasion. In an attempt to avoid it, we suggest to maintain the treatment of the underlying disease and wait for a minimum period of disease stability before the procedure.     

不同区域晶状体上皮细胞基因表达差异分析

【目的】比较不同区域猪晶状体上皮细胞的基因表达在转录组水平上的差异。【方法】解剖显微镜下分离晶状体前囊膜，将附着于其上的上皮细胞分为中央（直径为10．56mm）和周边两部分。分别提取两个样本的总RNAs并经PCR扩增，以Cy3和Cy5分别标记扩增的中央与周边部分的cDNA。与含7548个基因的表达谱芯片杂交，经图像分析，生物信息学处理获得基因表达在转录水平差异的相关信息。【结果】中央与周边区域的猪晶状体上皮细胞在转录组水平共鉴定出952个有效表达的基因点，其中差异表达基因261个．以中央区域为参照，周边上皮细胞mRNA上调137个，下调124个。差异表达基因主要涉及的功能有：细胞周期与凋亡、细胞骨架蛋白及细胞外基质、转录、细胞信号分子等。【结论】中央与周边区域猪晶状体上皮细胞基因表达在转录组水平上差异明显。这类差异呈明显的功能聚类。     

人食管不典型增生上皮与原位癌的计算机纹理分析研究

采用计算机图象纹理分析和相关点阵检测技术,对人食管正常粘膜、不典型增生上皮及原位癌的不同纹理特征进行了观察。观察样品为常规病理切片,用计算机图象分析系统检测了组织的纹理特征。对受检图象建立了三种灰色分层关系矩阵,同时计算了8种纹理测度。结果显示,在重度不典型增生上皮和原位癌之间,其纹理测度和相关点阵检测数据均有显著性差异(P<0.05)。全部测量数据经计算机多元逐步判别分析,其正判率达90%以上。本研究结果表明,计算机纹理分析方法可正确地判别食管癌前病变和原位癌的组织结构异型性。提示本技术在食管癌的早期诊断方面具有肯定的实用性价值。     

纤维蛋白胶体外重建兔角膜上皮的实验研究

目的利用组织工程技术体外重建角膜上皮组织,为重建角膜表面提供良好的移植材料和方法。方法以等量凝血酶和纤维蛋白原作用生成的纤维蛋白胶为载体(实验组),种植兔角膜缘组织块,体外培养重建角膜上皮层,每天记录细胞生长覆盖面积,从生长特性、光镜特征、超微结构特点以及免疫细胞化学方面进行观察检测。结果角膜缘上皮细胞在纤维蛋白胶上黏附生长增殖,体外培养7d左右即形成单层上皮细胞,2周左右形成的复层上皮细胞与纤维蛋白胶构成角膜上皮组织经检测与生理状态下的角膜上皮组织相近。实验组与对照组(置盖玻片为载体)比较细胞生长覆盖面积第4天至第7天差异有显著性(P<0.05)。结论以纤维蛋白胶为载体,体外培养的角膜上皮组织片可作为眼表重建角膜上皮移植良好的来源。     

Polarized expression of Na+/H+ exchange activity in LLC-PK1/PKE20 cells: II. Hormonal regulation

LLC-PK₁/PKE₂₀ cells (a continuous epithelial cell line) has two different Na/H exchange activities: Na/H-1 located in the basolateral membrane and Na/H-2 located in the apical membrane [Casavola et al. (1989) Biochem Biophys Res Commun 165:833–837; Haggerty et al. (1988) Proc Natl Acad Sci USA 86:6797–6801]. In the present report we have studied hormone regulation of these exchange activities by measuring Na-dependent recovery of pH_i from an acid load (by using microspectrofluorometry and 2,7-bis(carboxyethyl)-5,6-carboxyfluorescein) in response to activation of regulatory cascades by either pharmacological agents or by vasopressin or calcitonin. Agents leading to activation of protein kinase A (cAMP-dependent), such as forskolin (10 M), 8-Br-cAMP (0.25 mM), and isobutylmethylxanthine (0.5 mM), inhibited Na/H-2 and Na/H-1 by an average of 49%. Stimulation of protein kinase C by a phorbol ester (phorbol 12-myristate 13-acetate, TPA, 100 nM) inhibited Na/H-2 (by an average of 48%) and stimulated Na/H-1 (by an average of 38%); these effects of TPA were also observed in the presence of forskolin (100 M). Addition of either vasopressin (2 M) or calcitonin (0.3 M) onto both sides of the monolayer decreased the activity of Na/H-2 by an average of 26.3% and 27.7% respectively, and stimulated the activity of Na/H-1 by an average of 17.4% and 38.7% respectively; exposure of cells to either hormone stimulated production of cAMP and inositol trisphosphate, respectively. Separate hormone additions to either the apical or basolateral cell surface led to effects similar to those produced by simultaneous hormone additions onto both cell surfaces, although the relative response of Na/H exchangers to either agonist is variable. In summary, these results suggest that in LLC-PK ₁/PKE₂₀ cells, vasopressin and calcitonin can act via receptor systems coupled either to adenylate cyclase or to phospholipase C. Activation of these receptor systems can lead to inhibition of Na/H-2 and stimulation of Na/H-1.     

Melanocytes in the Testes of Eupemphix nattereri (Anura,Leiuperidae): Histological,Stereological, and Ultrastructural Aspects

Ectothermic vertebrates have a well‐developed system of melanin‐containing cells, which localize in several organs and tissues and compose an extracutaneous pigmentary system. This research aimed at characterizing histological and ultrastructural patterns of pigmented cells in the testes of the anura Eupemphix nattereri (Steindachner, 1963), including the stereological and quantitative evaluation of this cell type in the gonads. Ten adult males were collected in Nova Itapirema, São Paulo, Brazil, and submitted to morphological studies with light and transmission electron microscopy. The testis presents a great number of large cells with many brown granules and long cytoplasmic processes. The pigmented cells found in the testis are structurally similar to melanocytes, characterized by large amounts of melanosomes. The cells may be in intimate contact with the same cell type, with myoid cells surrounded by a large amount of collagen fibers, Leydig cells, and next to fibroblasts. The distribution and amount of extracutaneous melanocytes is variable when other organs and membranes are analyzed, allowing the establishment of species‐specific patterns for the extracutaneous pigmentary system. Anat Rec, 2007. © 2007 Wiley‐Liss, Inc.     

Properties and regulation of chloride channels in cystic fibrosis and normal airway cells

The present study examines the properties of Cl^–channels in cultured respiratory cells of cystic fibrosis (CF) patients and normal (N) individuals. In excised membrane patches the conductances for CF and N Cl^– channels were larger at positive as compared to negative clamp voltages (V _c): 74±2.6 (V _c > 0) and 47±2.0 pS (V _c < 0) for CF (n= 57) and 69±3.6 (V _c > 0) and 45±2.3 pS (V _c < 0) for N (n=35). The open probability (P _o) of the channel increased markedly with depolarization. Both the voltage dependence of the conductance and of P _o contribute to the outward rectification of the channel. The time histogram analysis reveals two open and two closed time constants. The selectivity of the channel was Cl^–=Br^– =I^– > NO ₃ ^– gluconate. The channel was inhibited reversibly by 5-nitro-2-(3-phenylpropylamino)-benzoate (NPPB) at 10^–7 mol/l to 10^–5 mol/l. While Cl^– channels were present in cell attached patches of N cells, they were absent in those of CF cells. The mean conductance for cell attached (N) Cl^– channels was 76±3.2 pS for positive clamp voltages (V _c) and 46±3.9 pS for negative V _c (n=8). When the membrane patches were excised from CF cells Cl^– currents appeared spontaneously (n=19). The immediate appearance (within 1 s) of Cl^– channels after excision was observed at positive (n=6) as well as at negative clamp voltage (n=13). Excision activation of CF Cl^– channels was observed at low (< 10^–9 mol/l) or high (10^–3 mol/l) calcium activities on the cytosolic side of the excised patch. Variation of the Ca⁺ activity (< 10^–9–10^–3 mol/l) or pH (6.5–8.5) on the cytosolic side exerted no effects on these Cl^– channels. These results suggest that Cl^– channels are present in the apical membrane of CF and N respiratory cells but they seem to be inhibited in intact CF cells. Excision of the patch and hence removal of the cytosolic inhibitor leads to an activation of Cl^– channels. The Cl^– channels in excised patches of N and CF cells have identical properties.     

Increased chloride permeability of amphibian epithelia treated with aldosterone

The transepithelial flux of chloride was increased by aldosterone treatment of amphibian skin and bladder and this was reflected by increased shunt conductance. The hormonal effect depended on the presence of chloride on the epithelial side of the preparation. These changes in tissue conductance and chloride permeability appear to be a direct effect of aldosterone as they did not occur when sodium transport was stimulated with vasopressin or hypotonicity.Chloride efflux was reduced in magnitude by indacrinone and DIDS, as well as after removal of chloride from the solution on the epithelial side of the preparations. These results suggest that, rather than merely diffusing along (a) paracellular pathway(s), chloride flows through (a) cellular structure(s), notably mitochondria-rich cells. These cells can therefore be considered as targets for aldosterone.Presented in part before the Belgian Physiological Society (Beauwens et al. 1985)     

Pigmentary degeneration indwed by N-methyl-N-nitrosourea and the fate of pigment epithelial cells in the rat retina

Pigmentary degeneration of the retina was induced by a single intraperitoneal Injection of 75mgkg of N-methyl-N-nitrosourea (MNU) In female Brown-Norway colored rats at 50 days of age, which were then observed at 24, 48 and 72 h and 7, 21,35 and 150 days after the treatment. MNU-treated rats showed selective destruction of the photoreceptor cells by an apoptotic mechanlsm 24 h after the treatment, and the destruction was completed by day 7. During the photoreceptor cell degeneration, proliferation of Miller cells and infiltratlon of macrophages was prominent 72h and 21 days aRttr the treatment, respectively. Müller cell proliferation and macrophage infiltratbn corresponded to degenerative photo-receptor cell phagocytosis, and prollferating Müller cell processes responded to stabilize the damaged retina. Pigment epithelial cell detachment from the Bruch's membrane was seen 72 h after the treatment, and migration within all layers of the retina was seen at day 7 when photoreceptor Cells were lost. At 21, 35 and 150 days after the treatment, lack of photoreceptor cells and deposition of pigment epithelial cells within the retina but not in contact to vascular endothe-lial cells were characteristic. MNU-induced photoreceptor apoptosis followed by Miiller cell and macrophage reaction then pigment epithellal cells deposition withln the retina partially resembles retinitis pigmentosa in humans.     

Regulatory volume decrease in a renal distal tubular cell line (A6) II. Effect of Na+ transport rate

A6 epithelia, a cell line originating from the distal tubular part of the kidney ofXenopus laevis, were cultured on permeable supports and mounted in an Ussing-type chamber. Cell thickness (T _c), short-circuit current (I _sc) and transepithelial conductance (G _t) were recorded while tissues were bilaterally incubated in NaCl solutions and the transepithelial potential was clamped to zero. Effects of inhibition and stimulation of transepithelial Na⁺ transport on cell volume and on its regulation during a hyposmotic challenge were investigated. Under control conditions a slow spontaneous decrease ofT _c described by a linear baseline was recorded. The reduction of the apical osmolality from 260 to 140 mosmol/kg did not alter cell volume significantly, demonstrating a negligible water permeability of the apical barrier. The inhibition of Na⁺ uptake by replacing apical Na⁺ byN-methyl-d-glucamine (NMDG⁺) did not affect cell volume under isotonic conditions. An increase ofT _c by 12.1% above the control baseline was recorded after blocking active transport with ouabain for 60 min. The activation of Na⁺ transport with insulin or oxytocin, which is known to activate the apical water permeability in other epithelia, did not alter cell volume significantly. The insensitivity of cell volume to alterations in apical Na⁺ uptake or Na⁺ pump rate confirms the close coupling between apical and basolateral transport processes. The blockage of basolateral K⁺ channels by 5 mM Ba²⁺ elicited a significant increase inT _c of 16.3% above control. Quinine, a potent blocker of volume-activated K⁺ channels, did not changeT _c significantly. Basolateral hypotonicity elicited a rapid rise inT _c followed by a regulatory volume decrease (RVD). An RVD was also recorded after blocking apical Na⁺ uptake as well as after stimulating apical Na⁺ uptake with oxytocin or insulin. Inhibition of active transport with ouabain as well as blocking K⁺ efflux at the basolateral side with Ba²⁺ or quinine abolished the RVD. The inhibition of the RVD by ouabain seems to be caused by a depletion of cellular K⁺, whereas the effects of Ba²⁺ and quinine are most likely due to the blockage of the basolateral K⁺ pathway.