色素内镜诊断早期胃癌组织和癌旁组织基因表达差异

目的探讨早期胃癌组织及癌旁组织ZO-1和ZO-2表达差异。方法将活检的早期胃癌组织和癌旁组织液氮冷冻保存后,分别进行RT-PCR、Western blot来观察ZO-1和ZO-2表达。结果ZO-1和ZO-2的表达广泛存在于人类胃癌和癌旁组织中,ZO-1和ZO-2在人类胃癌组织的表达量均较癌旁组织有显著下降。结论ZO-1和ZO-2可能与人类胃癌的发生和转移有密切的关系。     

腹腔镜胆囊切除术治疗胆囊颈结石嵌顿32例无并发症经验总结

目的总结LC治疗胆囊颈管嵌顿结石防止并发症发生的经验。方法回顾分析2010年2月至2011年1月32例胆囊颈管嵌顿结石患者行LC的临床资料。结果 32例均顺利完成LC,无1例胆管损伤、胆漏等手术并发症发生及死亡病例。术后住院3～9d,平均术后住院4d。术后随访6～11个月无并发症发生。结论为了减少并发症,应严格把握手术时机和适应证及手术技巧与技术要点,严格预防胆管损伤、胆漏、出血等并发症发生,必要时应及时中转开腹手术功。     

应用两种方法评价表皮神经的对比研究

目的 对比研究计数法与评分法在评价表皮神经中的价值.方法 使用计数法和评分法同时评价7名正常人和10例色素痣患者表皮神经.计数法:计数表皮神经纤维密度(IENFD),以单位表皮面积内的蛋白基因产物9.5(PGP 9.5)阳性神经纤维数表示.评分法:按照神经数量多少分别评为3、2、1、0分;同时观察神经形态好坏,分别评为3、2、1、0分,2项指标得分的平均值即为该标本的最终评分.结果 计数法评价正常人IENFD为550～1 125根/mm2,平均(589±236)根/mm2.应用评分法评价正常标本,7份标本的形态和数量评分都是3分.应用评分法评价10例色素痣表皮神经,最好的3例标本是6、10、4号;最差的3例标本是1、5、3号.应用计数法评价表皮神经最好的3例标本是6、10、8号;最差的3例标本是9、1、3号.有8例标本使用2种方法 得出的结果 是一致的,只有8、9号标本的结果 差异很大.2种方法 评价表皮神经的符合率为80%.结论 2种方法 均可作为表皮神经的评价方法 .在多数情况下,计数法较评分法更为精确;在少数情况下,评分法较计数法更能够客观地反映表皮神经的实际情况.     

人视网膜色素上皮细胞原代培养冻存和复苏方法改进

目的 优化建立一种简单有效的人视网膜色素上皮细胞(RPE)的原代培养、冻存和复苏的方法,对细胞进行鉴定,为研究RPE 的功能和治疗有关眼病提供细胞来源.方法 采用0.25%胰酶+0.02%乙二胺四乙酸(EDTA)二次消化分离细胞并进行传代,以0.125%胰酶进行传代,用免疫组织化学检测进行角蛋白与S100 表达鉴定RPE.利用-80 ℃冰箱对细胞冻存,复苏后观察细胞活性.结果 RPE 体外生长旺盛、1 h 贴壁.初期为圆型,胞壁内充满黑色色素,传代后细胞呈圆型、不规则形,细胞透亮.免疫组织化学角蛋白与S100 均呈强阳性,证实培养的是RPE 细胞,所获细胞纯度100%.冻存细胞复苏后生长旺盛,细胞形态与复苏前相比无区别.结论 二次胰蛋白酶消化分离细胞数量多,能成功建立RPE 的体外培养模式,用-80 ℃冻存RPE 的方法简便,成本低,冻存复苏后细胞存活率及细胞形态不受影响,是目前较理想的RPE 细胞保存方法.     

强脉冲光对烧伤深Ⅱ°创面愈合后瘢痕以及色素预防性护理

[目的]探讨强脉冲光对烧伤深Ⅱ°创面愈合后瘢痕以及色素预防性护理.[方法]选取2011年6月至2012年3月烧伤深Ⅱ °创面愈合患者60例,随机分为实验组和对照组.实验组采用强脉冲光加辅助性弹力压迫进行预防性护理,对照组仅采用辅助性弹力压迫.[结果]两组患者各种皮肤瘢痕类型的总有效率差异均有显著性(P＜0.05);两组患者的皮肤瘢痕色素沉着的总有效率有显著差异(90.00％vs 80.00％,P＜ 0.05).[结论]强脉冲光对烧伤深Ⅱ°创面愈合后瘢痕以及色素预防性护理效果佳、较安全,值得推广应用.     

Paclitaxel interrupts TGF-beta1 signaling between gallbladder epithelial cells and myofibroblasts

BACKGROUND: The cellular and molecular mechanisms of fibrogenesis in the extrahepatic biliary epithelium are not known. Transforming growth factor-beta1 (TGF-beta1) is a cytokine implicated in signaling pathways that mediate collagen formation. An observation that paclitaxel (PT), applied topically into the rat common bile duct, inhibited stricture formation led us to hypothesize that PT's effects might be due to interruption of TGF-beta1 signaling between biliary epithelial cells and subepithelial myofibroblasts. MATERIALS AND METHODS: We tested this hypothesis using an in vitro cell-culture model in which murine gallbladder epithelial cells (GBEC) are cultured separately or cocultured with human gallbladder myofibroblasts (GBMF). RESULTS: Exposure to Escherichia coli lipopolysaccharide (LPS) enhanced TGF-beta1 mRNA expression and stimulated TGF-beta1 protein secretion into both apical and basolateral compartments in GBEC. This effect was more prominent with basolateral secretion and was also more pronounced in the coculture system. In GBMF, collagen I mRNA expression and protein secretion were stimulated by treatment with LPS or TGF-beta1. GBMF also expressed TGF-beta1 mRNA, whose levels were enhanced by exposure to either LPS or exogenous TGF-beta1. PT inhibited LPS-induced TGF-beta1 mRNA expression and protein secretion in GBEC in both culture systems. Tumor necrosis factor-alpha mRNA expression and protein secretion were not affected by PT in GBEC, demonstrating that the effects were specific for TGF-beta1. PT also inhibited LPS- and TGF-beta1-induced collagen I mRNA expression and protein secretion in GBMF. CONCLUSIONS: These findings support a model in which GBEC communicate with subepithelial GBMF via TGF-beta1, leading to collagen deposition and fibrosis, and in which GBMF possess autocrine mechanisms involving TGF-beta1 that could regulate collagen production. PT inhibits these fibrogenic pathways.     

色素上皮衍生因子与肿瘤

色素上皮衍生因子(PEDF)是丝氨酸蛋白酶抑制剂(serpin)基因家族的成员,是一种强效的内源性血管生成抑制剂.PEDF在多种肿瘤组织中表达下调或缺失,与预后有很大相关性.PEDF的生物学特性、抗肿瘤活性机制的研究为其在肿瘤的治疗提供了理论基础.     

九里香叶色素的提取及其稳定性研究

研究九里香叶色素的提取条件和稳定性。以水作提取剂,在室温下浸提24 h,用紫外分析法研究色素的稳定性。色素在330 nm有最大吸收峰,该色素水溶性较好,pH值在1≤pH≤11对此色素的稳定性影响不是很大,但热稳定性良好。淀粉对九里香叶色素影响较大,苯甲酸钠、柠檬酸、醋酸、蔗糖、葡萄糖、维生素C对九里香叶色素稳定性无明显影响,耐光性较差。氧化剂、还原剂对色素的影响明显,其它均无明显影响,金属离子对其影响较小。     

Elucidating the phenomenon of HESC-derived RPE: anatomy of cell genesis, expansion and retinal transplantation

Healthy Retinal Pigment Epithelium (RPE) cells are required for proper visual function and the phenomenon of RPE derivation from Human Embryonic Stem Cells (HESC) holds great potential for the treatment of retinal diseases. However, little is known about formation, expansion and expression profile of RPE-like cells derived from HESC (HESC-RPE). By studying the genesis of pigmented foci we identified OTX1/2-positive cell types as potential HESC-RPE precursors. When pigmented foci were excised from culture, HESC-RPE expanded to form extensive monolayers, with pigmented cells at the leading edge assuming a precursor role: de-pigmenting, proliferating, expressing keratin 8 and subsequently re-differentiating. As they expanded and differentiated in vitro, HESC-RPE expressed markers of both developing and mature RPE cells which included OTX1/2, Pax6, PMEL17 and at low levels, RPE65. In vitro, without signals from a developing retinal environment, HESC-RPE could produce regular, polarised monolayers with developmentally important apical and basal features. Following transplantation of HESC-RPE into the degenerating retinal environment of Royal College of Surgeons (RCS) dystrophic rats, the cells survived in the subretinal space, where they maintained low levels of RPE65 expression and remained out of the cell cycle. The HESC-RPE cells responded to the in vivo environment by downregulating Pax6, while maintaining expression of other markers. The presence of rhodopsin-positive material within grafted HESC-RPE indicates that in the future, homogenous transplants of this cell type may be capable of supporting visual function following retinal dystrophy.