骨桥蛋白在不同分期糖尿病肾病患者肾组织中的表达

目的:探讨骨桥蛋白在不同分期糖尿病肾病患者肾组织的表达及其临床意义.方法:用免疫组化方法检测无糖尿病肾病组(n=20)、早期糖尿病肾病组(n=20)、临床糖尿病肾病组(n=20)肾组织骨桥蛋白的表达情况.结果:在不同分期糖尿病肾病组的骨桥蛋白表达不同,无糖尿病肾病组骨桥蛋白表达最低,临床糖尿病肾病组骨桥蛋白表达最高.结论:骨桥蛋白可能是糖尿病肾病的重要发病因素之一,抑制糖尿病患者肾脏骨桥蛋白的表达,可能延缓糖尿病肾病的进展.     

OPN、MMP-9在胰腺癌发展及侵袭转移中的意义

目的探索OPN、MMP-9在胰腺癌组织中的表达情况。方法采用免疫组织化学方法检测46例胰腺癌患者手术切除的正常胰腺组织和胰腺癌组织中的OPN、MMP-9的表达情况,并对不同肿瘤分化程度、临床分期及淋巴结转移的标本作对比分析。结果胰腺癌与正常组织之间OPN、MMP-9阳性表达率均有显著性差异（P〈0.05）,正常组织显著高于胰腺癌组织;胰腺癌与正常组织之间,两种大分子蛋白的表达与胰腺癌的组织分化程度、临床分期密切相关（P〈0.05）。有淋巴结转移的OPN的表达与无淋巴结转移的有显著性差异（P〈0.05）。结论 OPN、MMP-9两种大分子蛋白与胰腺癌的发生、发展密切相关。     

维生素K3对尿路结石病人尿中骨桥蛋白浓度的影响

目的：探讨维生素l（3对尿路结石病人尿中骨桥蛋白浓度的影响;从而观察维生素K3对尿路结石病人的治疗和预防复发的效果。方法：随机选取30例尿路结石患为维生素K组,肌注维生素K312mg／d,连续7天;收集用药前后的尿标本;随机选30例正常人做对照组,收集尿标本;利用westernblotting技术测定各尿标本中的骨桥蛋白含量;比较尿路结石患者用药前后及与正常人尿中骨桥蛋白含量的差别。统计学方法：配对资料均数t检验。结果：维生素K组用药后比用药前尿中骨桥蛋白含量高18．7％（P〈0．05）;用药前组比正常对照组尿中骨桥蛋白含量高93％,但差别无统计学意义（P=0．036）;用药后组比正常对照组尿中骨桥蛋白含量明显升高129％（P=0．0045,〈0．01）。结论：临床应用维生素K可上调肾骨桥蛋白的表达,抑制结石的形成,对尿路结石的治疗和预防有一定效果,值得推广。     

杜仲对大鼠骨髓基质细胞骨桥蛋白和骨保护素表达的影响:水提液与醇提液有区别吗？

背景：以往实验证实杜仲能促进骨髓基质细胞向成骨细胞分化,但对促进成骨分化的分子机制报道很少。 目的：观察中药杜仲水/醇提取物对大鼠骨髓基质细胞骨桥蛋白和骨保护素表达的影响 方法：盐杜仲2 g,分别加蒸馏水或甲醇至15 mL,室温振荡1 h,静置过夜后,15 000 r/min离心10 min,弃沉淀。水浸上清不经处理即得水提取物;甲醇浸上清经真空浓缩,再加水至15 mL溶解剩余物后,得醇提取物水、醇提取物,用0.22 μm滤膜过滤后于-20 ℃保存。取2月龄SD 大鼠6 只,体外培养至第3代SD大鼠骨髓基质细胞进行杜仲水/醇提取物诱导,诱导物分为1×10-2,1×10-3,1×10-4和1×10-5 4个稀释度;阴性对照组加入PBS;诱导培养6 d。以免疫细胞化学法测定诱导6 d的骨髓基质细胞骨桥蛋白和骨保护素表达。应用反转录-聚合酶链反应方法测定1×10-3,1×10-4稀释度诱导3 d的骨髓基质细胞骨桥蛋白和骨保护素的基因表达。 结果与结论：细胞免疫化学显示,杜仲水/醇提取物对大鼠骨髓基质细胞骨桥蛋白的表达均有显著的刺激作用,以 1×10-4的稀释度作用最强,且醇提取物的作用大于水提取物;骨保护素的表达却无显著变化。反转录-聚合酶链反应显示,水提取物对诱导3 d后大鼠骨髓基质细胞骨桥蛋白的基因表达量显著高于醇提取物,骨保护素未检测出表达。证实杜仲水/醇提取物诱导骨髓基质细胞成骨分化与刺激骨桥蛋白表达相关,与骨保护素无关;水提取物促进骨髓基质细胞骨桥蛋白表达早于醇提取物。     

骨桥蛋白在新生大鼠背部皮肤创伤愈合过程中的表达

目的：检测骨桥蛋白（osteopontin,OPN）在新生大鼠皮肤及创伤愈合过程中的表达,探讨OPN参与皮肤创伤愈合的可能机制。方法：在7d龄SD大鼠背部皮肤做直径为1ClTI的圆形全层皮肤切口,运用免疫组织化学方法检测OPN阳性细胞在皮肤及创伤愈合过程中的分布情况,并分别运用RT—PCR及Westernblot检测OPN在创伤愈合不同时间点（1、3、5及7d）的表达情况。结果：在大鼠正常皮肤,免疫组织化学结果显示OPN阳性细胞分布在毛囊、皮脂腺及汗腺,表皮则无表达;RT—PCR及Westernblot均未检测到OPN在正常皮肤中的表达。从创伤后第1天起,OPN阳性细胞开始集中分布在创伤边缘的真皮部位,且在第3天达到高峰,第、5天表达开始减弱。RT—PCR及Westemblot结果一致．OPN相对表达量在第3天达到高峰。结论：研究结果表明OPN参与了皮肤创伤愈合,尤其在创伤修复的炎症期及细胞外基质沉积期。     

Full length amelogenin binds to cell surface LAMP-1 on tooth root/periodontium associated cells

Objectives

Lysosome-associated membrane protein-1 (LAMP-1) has been suggested to be a cell surface receptor for a specific amelogenin isoform, leucine-rich amelogenin peptide or LRAP. However, it is unclear if LAMP-1 is an amelogenin receptor for dental mesenchymal cells. The goal of this study was to determine if LAMP-1 serves as a cell surface binding site for full length amelogenin on tooth root/periodontium associated mesenchymal cells.

Design

Murine dental follicle cells and cementoblasts (OCCM-30) were cultured for 2 days followed by addition of full length recombinant mouse amelogenin, rp(H)M180. Dose-response (0-100 μg/ml) and time course (0-120 min) assays were performed to determine the optimal conditions for live cell surface binding using immunofluorescent microscopy. A competitive binding assay was performed to determine binding specificity by adding Emdogain^® (1 mg/ml) to the media. An antibody against LAMP-1 was used to detect the location of LAMP-1 on the cell surface and the pattern was compared to cell surface bound amelogenin. Both amelogenin and cell surface LAMP-1 were immuno-co-localized to compare the amount and distribution pattern.

Results

Maximum surface binding was achieved with 50 μg/ml of rp(H)M180 for 120 min. This binding was specific as demonstrated by competitive inhibition (79% lower) with the addition of Emdogain^®. The binding pattern for rp(H)M180 was similar to the distribution of surface LAMP-1 on dental follicle cells and cementoblasts. The high co-localization coefficient (0.92) for rp(H)M180 and LAMP-1 supports rp(H)M180 binding to cell surface LAMP-1.

Conclusions

The data from this study suggest that LAMP-1 can serve as a cell surface binding site for amelogenin on dental follicle cells and cementoblasts.     

康莱特对人结肠癌裸鼠种植瘤肝转移的影响

目的：探讨中药康莱特对人结肠癌裸鼠种植瘤肝转移的抑制作用。方法：将结肠癌LoVo细胞注入裸鼠脾脏,建立结肠癌肝转移模型。将成功制模的裸鼠随机分为4组：对照组,CTX组,康莱特组及CTX＋康莱特组。治疗45天后,处死裸鼠,计算肝转移结节数。采用ELISA法测定肿瘤组织上清液骨桥蛋白（OPN）的浓度。结果：在肝转移结节数目上,与对照组相比其余3组肝转移结节个数均降低且有统计学意义。在组织骨桥蛋白浓度上,其余3组组织骨桥蛋白浓度降低且有统计学意义。与CTX组相比,CTX＋康莱特组组织骨桥蛋白浓度降低且有统计学意义,但CTX组与康莱特组二者无统计学差异P=0.176。结论：康莱特可以抑制裸鼠结肠癌的肝转移,康莱特联合CTX具有协同作用。     

血清骨桥蛋白和CA125联合检测鉴别良恶性腹水

目的:探讨联合检测血清骨桥蛋白(OPN)和CA125在良恶性腹水中的诊断价值。方法:取39例恶性腹水患者、48例结核性腹水患者及40例健康妇女血清样品,分别用双抗体夹心酶联免疫吸附法(ELISA)及微粒子化学发光法测定其OPN和CA125水平,观察其在诊断良、恶性腹水时的敏感性和特异性。结果:恶性腹水患者血清OPN水平明显高于结核性腹水患者及健康妇女组。OPN检测恶性腹水的敏感度、特异度分别为84.6%、95.0%;CA125检测敏感度、特异度为76.9%、87.5%;联合检测敏感度97.4%、特异度为95.0%。联合检测OPN、CA125较单独检测OPN及CA125其敏感度高(P<0.05);特异度无明显差异。结论:OPN与CA125联合测定是鉴别卵巢癌和结核性腹水的有效方法,联合检测CA125可提高其临床应用价值。     

TGFBR2 but not SPP1 genotype modulates osteopontin expression in Duchenne muscular dystrophy muscle

A polymorphism (rs28357094) in the promoter region of the SPP1 gene coding for osteopontin (OPN) is a strong determinant of disease severity in Duchenne muscular dystrophy (DMD). The rare G allele of rs28357094 alters gene promoter function and reduces mRNA expression in transfected HeLa cells. To dissect the molecular mechanisms of increased disease severity associated with the G allele, we characterized SPP1 mRNA and protein in DMD muscle biopsies of patients with defined rs28357094 genotype. We did not find significant differences in osteopontin mRNA or protein expression between patients carrying the T (ancestral allele) or TG/GG genotypes at rs28357094. The G allele was significantly associated with reduced CD4⁺ and CD68⁺ cells on patient muscle biopsy. We also quantified transforming growth factor‐β (TGFB) and TGFB receptor‐2 (TGFBR2) mRNA in DMD muscle biopsies, given the ability of TGFB and TGFBR2 to activate SPP1 promoter region and their role in DMD pathogenesis. The amount of TGFB and TGFBR2 mRNA did not predict the amount of SPP1 mRNA or protein, while a polymorphism in the TGFBR2 gene (rs4522809) was found to be a strong predictor of SPP1 mRNA level. Our findings suggest that OPN mediates inflammatory changes in DMD and that TGFB signalling has a role in the complex regulation of osteopontin expression. Copyright © 2012 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.