激光上皮下角膜磨镶术后角膜上皮瓣临床观察

目的 观察及探讨准分子激光上皮下角膜磨镶术(Laser subepithel ialkeratomileusis，LASEK)后，角膜上皮瓣的成活率及其影响因素。方法 对行LASEK治疗的42例(80眼)于术后1、2、3天，1、2、3、4周在裂隙灯显微镜下进行角膜上皮瓣的观察。结果 34例(68眼)，角膜上皮瓣成活，成活占85％(68／80)；未成活8例(12眼)，未成活占15％(12／80)。结论 LASEK术后角膜上皮瓣成活率的高低，决定着LASEK的临床疗效，影响其成活的因素是多方面的。其中角膜上皮瓣边缘不整齐、破裂、对位不良、操作时间过长可能是其主要原因。     

枸橼酸减轻草酸钙晶体对大鼠肾小管上皮细胞损伤作用的实验研究

目的 通过研究枸橼酸能否减轻一水草酸钙（calcium oxalate monohydrate，COM）晶体对体外培养的Wistar大鼠肾小管上皮细胞造成的损伤，以探讨枸橼酸在尿路结石防治中的作用及其机制。方法分离、培养正常雄性Wistar大鼠的肾小管上皮细胞，在原代培养第5天时，将细胞分成3组（空白对照组、COM组及枸橼酸组），分别于各组培养液中培养2h后用比色法测定各组细胞培养基中丙二醛（malonaldehyde，MDA）的含量及细胞的钙镁ATP酶（Ca`2＋-Mg`2＋-ATPase）和钠钾ATP酶（Na`2＋K`＋-ATPase）的活性。结果①COM组和枸橼酸组细胞Ca2＋-Mg`2＋ATPase和Na`＋＋K`＋-ATPase活性均明显低于对照组（P〈0．05）；②与枸橼酸组相比，COM组细胞Ca`2＋-Mg`2＋-ATPase和a`＋＋K`＋-ATPase活性下降更加明显（P〈O．05）；③COM组和枸橼酸组细胞培养基中MDA的含量均明显高于对照组（P〈O．05）；④与枸橼酸组相比，COM组细胞培养基中MDA的含量更高（P〈O．05）。结论外源性枸橼酸能够减轻一水草酸钙晶体对体外培养的Wistar大鼠肾小管上皮细胞造成的脂质过氧化损伤。     

Expression of β-catenin in normal breast tissue and breast carcinoma: a comparative study with epithelial cadherin and α-catenin

Expression of β-catenin was investigated in normal breast tissue and 66 breast carcinomas in conjunction with expression of epithelial cadherin (E-CD) and α-catenin. In normal mammary ducts and acini, intense β-catenin immunoreactivity was present at the basolateral surfaces of luminal epithelium and weak immunoreactivity was observed at the lateral borders of myoepithelial cells. No β-catenin was revealed at the myoepithelial basal surface. The intercellular expression of β-catenin, as well as of E-CD and α-catenin, was also observed in carcinoma tissues with varying staining intensity. Almost all of 10 intraductal carcinomas and approximately 70% of 41 invasive ductal carcinomas expressed the three molecules at the same level as in normal glands, whereas approximately 80% of 13 invasive lobular carcinomas showed severe deficiency of them. Two lobular carcinomas in situ showed complete absence of all of the proteins. Some of these findings were confirmed biochemically by immunoblotting analysis. In invasive ductal carcinomas, α-catenin was reduced more frequently in diffuse than in solid type tumours, whereas the level of expression of β-catenin and E-CD was unchanged between them. No correlation was present between reduced expression of the adhesion molecules and lymph node metastasis.     

人腭扁桃体隐窝上皮结构的研究

采用扫描电镜、透射电镜及光镜免疫金银法观察了31例慢性扁桃体炎患者和8例胎儿的扁桃体隐窝上皮结构及其中细胞的分布。电镜观察发现隐窝上皮表面存在三种类型微隐窝开口,其腔内有浸润细胞、异物及细菌。有三种特化上皮细胞(M细胞)覆盖于Ⅲ型微隐窝开口处,其结构与肠道M细胞相似,其数量随扁桃体炎的反复发作而减少。形态表明微隐窝是浸润细胞和外来抗原的出入口。M细胞与抗原的摄取及传递有关。光镜免疫金银法观察证明上皮浸润细胞中多数为OKT_s~+细胞,其中OKT_4~+者又占多数而OKT_s~+细胞较少,这些细胞是隐窝上皮参于免疫应答的结构基础。     

Effects of topical anti-inflammatory drugs on eosinophil survival primed by epithelial cells. Additive effect of glucocorticoids and nedocromil sodium

Background Eosinophil infiltration is a hallmark of the inflammatory response in rhinitis and in nasal polypcsis. Objective We studied the effect of steroids and nedocromil sodium on eosinophil survival primed by epithelial cells from healthy (nasal mucosa) and inflamed (nasal polyp) respiratory tissue. Methods Blood eosinophils were incubated with increasing concentrations (10^-11 10^-5 M) of topical steroids (fiuticasone propionate, budesonide, triamcinolone acetonide and beclomethasone dipropionate) and/or nedocromil sodium prior to the addition of human epithelial cell conditioned media (HECM), eosinophil viability was measured and IC₅₀ for each drug was calculated. Results All four steroids and nedocromil sodium caused a dose-related inhibition of HECM-induced eosinophil survival. The IC₅₀ of steroids were lower in eosinophils primed by mucosa HECM than on those primed by polyp HECM (fluticasone, 4nM vs 114nM: budesonide, 21 nM vs 280 nM; triamcinolone, 7 nM vs 853 nM; and beclomethasone, 171 nM vs 181 nM). The combined inhibitory effect of 10^-7M budesonide plus 10^-5M nedocromil (43.8 ± 10.8%, P < 0.03) was significantly higher than budesonide (28.5 ± 9.2%) or nedocromil (16.7 ± 5.4%) alone and close to 10^-5M budesonide (52.3 ± 11%). No differences were found in cytokine (IL-8, IL-6, GM-CSF, TNFα, IL-lβ and RANTES) concentrations between HECM from mucosa and polyps. Conclusion These results suggest that topical anti-inflammatory drugs may diminish airway eosinophilic infiltration by decreasing eosinophil viability, that nasal polyp epithelial cell secretions may induce steroid resistance in eosinophils, and that nedocromil sodium has additive effects with steroids.     

Kinetic study of glomerular epithelial cells associated with segmental glomerular sclerotic lesions with adhesion in spontaneously diabetic WBN/Kob rats

Background We previously found that glomerular epithelial cells play an important role in the formation of adhesive lesions. Glomerular sclerotic lesions develop after the inital adhesive lesions. Methods Two series of experiments were done with spontaneously diabetic WBN/Kob rats. These rats develop segmental glomerular sclerotic lesions with aging. The first series of experiments was intended to clarify the kinetics of glomerular cells on progressive glomerular damage in these rats. The second series of experiments was designed to study the relationship between proliferation (judged by % bromodeoxyuridine-positive cells) of glomerlar epithelial cells and sclerotic lesions with adhesions. Results In the first series, rats having increased proteinuria showed segmental glomerular sclerotic lesions with adhesions. At the same time, increased labeling indices of tuft cells and epithelial cells of Bowman's capsule were observed. In the second series, no significant increase in the labeling indices of tuft cells with sclerotic lesions was observed, compared to tuft cells without sclerotic lesions. In sclerotic lesions with adhesion, bromodeoxyurdine-positive cells were observed that were not distinguishable as podocytes or epithelial cells of Bowman's capsule. The highest labelling index was noted in the epithelial cells of Bowman's capsules with sclerosis. Conclusion This study shows that the proliferation of glomerular epithelial cells (mainly epithelial cells of Bowman's capsule) occurs in glomerular sclerotic lesions with adhesions.     

羊膜负载骨髓间充质干细胞与表皮细胞对放创性皮肤损伤促愈合研究

目的探讨治疗放创性全厚皮肤缺损创面的方法及效果. 方法贵州小香猪8只,每只背部脊柱两侧均有放创性全层皮肤缺损圆形创面(Ф3.67cm)各3个,共48个创面.将经处理的人羊膜(human amniotic mambrane, HAM)分别负载自体骨髓间充质干细胞(mesenchymal stem cells, MSCs)和表皮细胞,移植到其左侧24个创面作为实验组(A组);以单纯无种植细胞的HAM敷盖其右侧前16个创面(B组);以单纯油纱布敷盖其右侧后8个创面(C组).B、C作为对照组.观察移植后1～3周内各组创面愈合、肉芽组织生长及上皮化等情况,并进行创面组织HE染色及vWF免疫组织化学检测.用图像分析法测算各组各时间点创面平均面积(cm2),并计算其愈合百分率. 结果 C组于伤后 22～23天愈合,B组于伤后19～21天愈合;A组于伤后15～17天愈合,较B、C组分别提前6～7天和5～6天,愈合质量好.移植15～17天,A组与B、C组创面平均残留面积及愈合面积百分率比较,差异有统计学意义(P＜0.01). A组创面的新生上皮已完全覆盖整个创面,肉芽组织生长旺盛,肉芽组织中vWF、成纤维细胞和毛细血管含量丰富,可见胶原沉积;B、C组创面仍见许多炎性细胞浸润,肉芽组织中vWF、成纤维细胞和毛细血管含量少,胶原沉积不明显. 结论 HAM负载自体MSCs和表皮细胞植入对放创性全厚皮肤缺损创面有较好的促愈合作用,愈合质量较高.     

人骨髓间充质干细胞向多巴胺神经元分化的体外研究

目的探讨人骨髓间充质干细胞(hMSC)向神经元和多巴胺神经元分化的潜能。方法分离和纯化hMSCs;在体外以WHI-P131预处理和碱性成纤维细胞生长因子预诱导后,全反式维甲酸和胶质细胞源性神经营养因子联合诱导hMSCs向神经元和多巴胺神经元分化。光镜下观察其分化过程中hMSCs的形态变化,免疫组化检测诱导前后细胞是否表达神经元和多巴胺能神经元标志蛋白。结果诱导后的hMSCs能分化成为具有典型神经元形态的细胞,并明显表达抗人神经巢蛋白(nestin)[(54.2±3.7)%]和神经元特异性烯醇化酶(NSE)[(77.0±5.7)%],低表达胶质纤维酸性蛋白(GFAP)[(8.8±2.4)%];对照组细胞这些表达均为阴性;而且相当部分hMSCs表达酪氨酸羟化酶(TH)[(36.5±15.8)%]和多巴胺转运体(DAT)[(26.0±14.2)%]。结论在适当条件下,hMSCs可分化成为神经元样细胞和多巴胺神经元样细胞。     

VEGF基因体外转染大鼠骨髓间充质干细胞的实验研究

目的:探讨脂质体介导血管内皮细胞生长因子(VEGF)基因转染大鼠骨髓间充质干细胞(MSCS)应用于基因治疗的可行性、安全性。方法:体外分离、培养、鉴定MSCs,PcDNA3.1(-)/VEGF165质粒转染MSCs,转染后用免疫荧光和ELISA检测MSCs表达VEGF蛋白的情况,MTT检测MSCs对VEGF质粒转染的敏感性。结果:骨髓中分离得到MSCs,流式细胞检测显示MSCs不表达CD34和CD45,但表达CD90。透射电镜观察可见细胞浆中含大量粗面内质网和分泌颗粒。VEGF基因转染MSCs后第5天抗VEGF免疫荧光染色约90%的MSCs呈阳性,ELISA检测结果显示PcDNA3.1(-)/VEGF165质粒转染组细胞培养上清液中VEGF含量明显高于对照组,并于转染后第5天达到峰值。MTT检测结果显示VEGF质粒转染对MSCs增殖无影响。结论:MSC可作为VEGF基因转染的靶细胞用于基因治疗。