正畸力作用下大鼠牙周组织中骨桥蛋白的表达

目的：观察SD大鼠正畸牙齿移动过程中牙周组织内骨桥蛋白（osteopontin,OPN）的表达,探讨OPN在正畸牙齿移动过程及成骨方面的作用和机制。方法：选用6～8周龄雄性SD大鼠40只,以左侧上颌第一磨牙为实验组,右侧上颌第一磨牙不加力为对照组,通过自制的加力装置牵引移动大鼠的磨牙,分别在加力后8h、24h、3d、7d处死实验动物,即刻取上颌骨制备组织标本,通过免疫组织化学方法检测SD大鼠牙周组织中OPN的表达。结果：实验组大鼠牙周膜中张力侧OPN表达明显高于对照组,3d、7d组差异有统计学意义（P〈0.01）。结论：在正畸治疗牙齿移动过程中,OPN参与牙周组织的改建并与新骨的形成相关。     

复方氯己定含漱液对种植义齿后种植体周围龈沟液中骨桥蛋白和碱性磷酸酶水平的影响

目的：探讨种植义齿后复方氯己定含漱液含漱对种植体周围龈沟液中骨桥蛋白（OPN）和碱性磷酸酶（ALP）水平的影响。方法：20例种植义齿患者随机分为A、B两组,每组10例,每位患者种植义齿1颗。A组患者义齿种植术后应用0.12%复方氯己定含漱液漱口,每次15 ml,早晚刷牙后含漱。B组患者只于早晚刷牙后清水漱口。两组均在种植术前2h及种植术后1.5个月采集种植体龈沟液1次,采用Western-boltting法检测OPN的表达,酶联免疫吸附法（ELISA）检测ALP水平。结果：两组患者种植义齿前OPN和ALP的表达差异无显著性（P〉0.05）,种植术后A组OPN和ALP的水平明显低于B组术后,差异有统计学意义（P〈0.05）。结论：种植义齿术后用复方氯己定含漱液含漱能够明显降低种植体周围龈沟液中OPN及ALP表达,有利于种植牙的成功种植。     

Pre-clinical in vitro and in vivo safety evaluation of bovine whey derived osteopontin,Lacprodan® OPN-10

Lacprodan® OPN-10 is a proprietary whey-based protein product that contains bovine-derived osteopontin (OPN), found in human milk and other bodily tissues. In vitro genotoxicity tests conducted according to accepted guidelines at up to 5000 μg/plate OPN failed to induce genetic mutations in Salmonella typhimurium strains and Escherichia coli strain and did not induce chromosomal aberrations or cytotoxicity in human lymphocytes. Administration of an acute dose of Lacprodan® OPN-10 (2300 mg/kg body weight) to male and female mice did not induce chromosomal damage or mitotic apparatus damage to erythroblasts from bone marrow. Lacprodan® OPN-10 was evaluated in a 13-week oral toxicity study in which rats were fed diets containing 0.5%, 1.0% and 2.0% Lacprodan® OPN-10. No test-article-related clinical observations or toxicological effects on body or organ weights, food consumption, ophthalmic effects, locomotor activity, hematology, clinical chemistry, urinalysis, or pathology were identified. In a teratogenicity study, administration of Lacprodan® OPN-10 up to 2500 mg/kg bw/day via gavage to pregnant rats had no effect on dams or pups. The No Observed Adverse Effect Level (NOAEL) for Lacprodan® OPN-10 in the 13-week toxicity study was 2.0% of the diet (equivalent to 1208 mg/kg bw/day in male rats and 1272 mg/kg bw/day in female rats).     

凉血养阴法对寻常型银屑病患者外周血VEGF和OPN表达的影响

目的 观察凉血养阴法对寻常型银屑病患者的临床疗效及外周血血管内皮细胞生长因子(VEGF)和骨桥蛋白(OPN)表达的影响,探讨凉血养阴法治疗银屑病的作用机制.方法 选取70例寻常型银屑病患者,随机分为治疗组(银屑平丸)和对照组(复方青黛丸)各35例.分别予以口服银屑平丸和复方青黛丸,疗程均为8周,同时设立20名健康体检者作对照.观察2组患者治疗前后的银屑病面积及严重程度指数(PASI)评分,同时采用酶联免疫吸附法(ELISA)检测2组患者治疗前后外周血VEGF和OPN含量变化,并与相应的PASI评分作相关性分析.结果 2组治疗后PASI积分均有明显下降,治疗组下降较对照组明显;治疗组总有效率为85.71％,对照组总有效率为58.82％,治疗组疗效优于对照组;2组患者治疗前外周血VEGF和OPN含量明显高于正常人,治疗后2组外周血VEGF和OPN均有下降,且治疗组较对照组下降更明显;银屑病外周血VEGF和OPN与其相应PASI呈正相关,但VEGF和OPN无明显相关性.结论 凉血养阴法治疗寻常型银屑病安全,疗效确切;寻常型银屑病患者外周血中VEGF和OPN表达异常,银屑平丸纠正异常表达的VEGF和OPN的水平,从而达到治疗银屑病的目的.     

依达拉奉对实验性自身免疫性脑脊髓炎大鼠的保护作用

目的观察依达拉奉对实验性自身免疫性脑脊髓炎大鼠的保护作用。方法将40只健康成年Wistar大鼠随机分成正常组、EAE组、LDE组、HDE组各10只,EAE组、LDE组、HDE组建立EAE模型,LDE组、HDE组分别腹腔注射相应剂量的药物。正常组、EAE组仅腹腔注射生理盐水0.5ml/d,所有大鼠24h后处死。结果EAE组神经功能缺损评分>LDE组>HDE组,组间比较有显著性差异(P<0.05);EAE组iNOS、OPN阳性细胞表达>LDE组>HDE组,组间比较有显著性差异(P<0.05)。结论依达拉奉对EAE的保护作用可能与其清除自由基、减轻炎症反应、减少i-NOS和OPN表达有关。     

脂多糖作用下腹膜间皮细胞骨桥蛋白表达及乌司他丁的影响

目的：观察脂多糖对大鼠腹膜间皮细胞（RPMC）中骨桥蛋白（OPN）表达的影响;观察乌司他丁（ulinastatin）作用下RPMC的OPN细胞分布及mRNA表达的影响。方法：胰蛋白酶法行RPMC的原代培养和传代,经鉴定第3代细胞融合至80%时分组。正常对照组：不同浓度LPS（50,100μg/ml）组;50μg/mlLPS作用不同时间（8,12,24,34,48h）组;乌司他丁组：（160,320,640U/ml）与LPS50mg/L作用24h,RealTime-RT-PCR技术检测骨桥蛋白（OPN）、TGF-β1mRNA的表达。结果：LPS可刺激RPMC的骨桥蛋白表达显著增加,呈剂量、时间依赖性（P〈0.05）;乌司他丁可降低LPS引起的RPMC中的骨桥蛋白表达,呈剂量依赖性（P〈0.05）。结论：脂多糖通过上调骨桥蛋白的表达,引起腹膜损伤导致腹膜纤维化,乌司他丁可以一定程度抑制骨桥蛋白引起的腹膜纤维化过程。     

Difference of osteopontin gene regulation between bone and kidney

 Osteopontin is a sialoprotein that is expressed in various cells. It plays a variety of important roles in cell adhesion, migration, signaling, calcification, and immunity. Its diverse functions indicate that the regulation of osteopontin may also vary extensively among tissues. Although osteopontin promoter has been studied in vitro, in vivo analyses may be more appropriate for elucidating osteopontin's functions. In an attempt to investigate osteopontin gene expression, we generated transgenic mice in which the bacterial β-galactosidase reporter gene was conjugated downstream of osteopontin promoter. The osteopontin promoter was a mouse −910 bp upstream fragment, which we had previously found functional in 3T3 cells. Among 34 transgenic founders, 13 mice were transgenic, as determined with the polymerase chain reaction. Osteopontin and β-galactosidase signals were evaluated with in situ hybridization. Among the 13 transgenic mice, 3 were β-galactosidase-positive. In these transgenic mice, osteopontin signals were observed in bones and kidneys, whereas β-galactosidase message was detected only in bones. This suggests that the −910 bp osteopontin promoter is active in bones but not in kidneys. These data imply that the promoter region required for osteopontin expression in kidneys may differ from that in bones. Received: March 22, 2002 / Accepted: December 5, 2002 RID="*" ID="*" Offprint requests to: T. Sakuma Acknowledgments. We greatly appreciate the excellent histotechnological assistance of Mr. Kenji Morihana. This work was supported in part by a grant from The Ministry of Education, Science, and Culture, Japan (no. 12470056).     

膝骨关节炎患者关节滑液中骨桥蛋白和软骨寡聚基质蛋白水平与疾病严重程度相关性

目的:探讨膝骨关节炎（knee osteoarthritis,KOA）患者关节滑液中骨桥蛋白（osteopontin,OPN）和软骨寡聚基质蛋白（cartilage oligomeric matrix protein,COMP）水平与疾病严重程度的相关性。方法：选取2018年2月至2020年5月收治的59例KOA患者作为KOA组,其中男25例,女34例;年龄60~75（65.57±1.56）岁;体质量指数（body mass index,BMI）21.4~30.7（26.12±1.54） kg/m²。采用Kellgren-Lawrence（K-L）分级对X线结果进行评估,其中Ⅱ级（K-L2组）14例,Ⅲ级（K-L3组）27例,Ⅳ级（K-L4组）18例。另选取18例因韧带或半月板疾病进行关节镜检查且无软骨损伤患者作为对照组,男7例,女11例;年龄61~78（64.88±1.60）岁;BMI 22.8~29.9（25.89±1.49） kg/m²。治疗前采集研究对象关节滑液样本,采用酶联免疫吸附试验检测关节滑液OPN、COMP水平,比较KOA组与对照组关节滑液OPN、COMP水平。比较不同K-L分级KOA患者性别、年龄、BMI等临床资料,采用酶联免疫试验检测其关节滑液中白细胞介素-1β（interleukin -1β,IL-1β）,OPN,COMP,基质金属蛋白酶3（matrix metalloproteinase -3,MMP-3）水平,比较不同K-L分级KOA患者临床资料和生化指标,采用Logistic回归分析影响KOA患者K-L分级的因素,采用ROC曲线下面积（area under the curve,AUC）预测KOA疾病严重程度。结果：59例KOA患者获得随访,时间8~27（15.75±3.27）个月。KOA组关节滑液OPN、COMP水平高于对照组（P<0.001）。K-L2组、K-L3组、K-L4组IL-1β、OPN、COMP、MMP-3水平比较差异有统计学意义（P<0.001）;与K-L2组比较,K-L3、K-L4关节滑液关节滑液IL-1β、OPN、COMP、MMP-3 水平均升高（P<0.05）;与K-L3组比较,K-L4关节滑IL-1β、OPN、COMP、MMP-3水平均升高（P<0.05）。多因素Logistic回归分析结果显示：关节滑液OPN、COMP、MMP-3水平是影响KOA患者K-L分级的独立危险因素（OR=6.653, 4.229,1.579,P<0.001）。关节滑液OPN预测K-L4级KOA的AUC为0.720[95%CI（0.588-0.851）],灵敏度为94.4%,特异度为65.9%;关节滑液COMP预测K-L4级KOA的AUC为0.731[95%CI（0.592-0.870）],灵敏度为88.9%,特异度为63.4%;关节滑液OPN联合COMP预测K-L4级KOA的AUC为0.839[95%CI（0.724-0.953）],灵敏度为94.4%,特异度为51.2%;OPN联合COMP预测K-L4级KOA的AUC大于单独OPN、COMP的AUC（Z=4.037,3.540,P<0.05）。结论：KOA患者关节滑液OPN、COMP水平升高,并随着K-L分级增加而升高。关节滑液OPN、COMP是影响KOA患者K-L分级的独立危险因素,二者预测K-L4级KOA 的AUC、灵敏度、特异度高,可用于评估KOA疾病进展。     

骨桥蛋白和P21在原发性肝癌中的表达及其临床意义

目的　探讨骨桥蛋白(OPN)和P2 1在原发性肝癌组织中的表达及它们在肝癌转移中的作用。方法　采用免疫组化S P法检测69例原发性肝癌组织中OPN和P2 1的表达情况。结果　①OPN和P2 1的阳性表达主要定位于癌细胞的细胞浆;OPN、P2 1在69例肝癌组织中阳性表达率分别为72 .5 % (5 0 / 69)和62 .3 % (4 3 / 69)。②在发生转移的肝癌组织中OPN和P2 1阳性表达率分别为84.2 % (3 2 / 3 8)和76.3 % (2 9/ 3 8) ,高于无转移的肝癌组织中OPN(5 8.1% ,18/ 3 1)和P2 1(4 5 .2 % ,14 / 3 1)阳性表达率,差异具有显著性,P <0 .0 5。③两者均阳性表达的3 5例肝癌中有2 4例(74.3 % )已发生转移,与两者均阴性表达的11例肝癌中仅有1例(9.1% )发生转移比较差异有显著性,P <0 .0 5 ,提示OPN和P2 1均阳性表达与肿瘤转移的发生密切相关。④OPN和P2 1的表达与肿瘤的大小、AFP水平、有无HBV感染、肿瘤分化、性别、年龄等因素无关,但与肿瘤侵袭转移的发生有关,P <0 .0 5。⑤OPN和P2 1的表达呈显著正相关(r =0 .2 5 7,P <0 .0 5 )。结论　OPN和P2 1的表达与肝癌侵袭转移的发生有密切关系,两者均阳性表达时对预测肝癌已发生转移具有指导意义。