Hypophosphatasia (HPP) is an inherited skeletal disease caused by mutation of the gene encoding tissue non-specific alkaline phosphatase (TNSALP). Odonto-HPP is well known as the mildest of HPP. The manifestations inv...Hypophosphatasia (HPP) is an inherited skeletal disease caused by mutation of the gene encoding tissue non-specific alkaline phosphatase (TNSALP). Odonto-HPP is well known as the mildest of HPP. The manifestations involve only the teeth, such as premature primary teeth exfoliation caused by reduction of alveolar bone, enlarged dental pulp chamber, and dental defects. We report a case of a 9-years-old boy who developed HPP. He was observed from the primary dentition to the mixed dentition period. At initial presentation at our hospital, he had multiple premature exfoliation of primary teeth and reduction of the alveolar bone. HPP was suspected due to the low level of ALP activity in serum, his oral manifestation, and dental history. He was referred to a physician for the final diagnosis. Therefore his compound heterozygote mutations, c.1559 delT (T/delT) and c.407G > A (G/A), were found in TNSALP and he diagnosed with odonto-HPP. Even though these mutations were reported as being involved in odonto-HPP, his mineral densities tended to be lower than that of his age. It is therefore necessary to investigate the bone mineralization level in odonto-HPP without other bone symptoms. Moreover, ongoing enzyme-replacement therapy in odonto-HPP might improve dental abnormality and bone disorders.展开更多
Background: Alkaline phosphatase has 4 isozymes, tissue non-specific alkaline phosphatase (TNAP), placental alkaline phosphatase (PLAP), intestinal alkaline phosphatase and germ-cell alkaline phosphatase. Hypophosphat...Background: Alkaline phosphatase has 4 isozymes, tissue non-specific alkaline phosphatase (TNAP), placental alkaline phosphatase (PLAP), intestinal alkaline phosphatase and germ-cell alkaline phosphatase. Hypophosphatasia (HPP) is an inherited skeletal disease caused by mutations of the gene encoding TNAP. Although TNAP is expressed in various tissues, the primary HPP symptoms appear in bones and teeth. The clinical severity of HPP varies widely from the most severe (perinatal, infantile and childhood) to the mildest forms (adult, and odonto-hypophosphatasia). We reported that gene therapy using a single injection of lentiviral vector expressing bone-targeted TNAP (TNAP-D10) is effective in preventing all the skeletal of HPP in TNAP knockout (Alpl–/–) mice as the model of infantile HPP. Objective: In this study we focus on evaluating the efficacy of treatment with gene therapy on the bone and teeth using TNAP-D10 and also we investigate the feasibility of gene therapy using bone-targeted PLAP (PLAP-D10). Methods and Findings: We used Alpl–/–mice that develop skeletal disease at postnatal days 6-8 mimicking the infantile form of human HPP. We injected 100 μl of lentiviral vectors harboring TNALP-D10 (5.0 × 107 TU) or PLAP-D10 (5.0 × 107 TU) to 1-day-old Alpl–/–?mice via the jugular vein. We performed histological analysis and micro-CT evaluation on bone and mandible of Alpl–/–?mice. The alveolar bone, enamel and dentin defects were corrected on treated Alpl–/–?mice by this treatment. Additionally the long bone growth rates (LGR) of long bones were encouraged on treated Alpl–/–?mice compared with untreated mice. Conclusions: These results indicate that the bone-targeted TNAP treatment mediated by lentivirus can correct not only the bone disorder but also the dental symptoms in Alpl–/–. This study also shows that PLAP-D10 can potentially be used to correct HPP disease.展开更多
目的探讨Preptin在2型糖尿病性骨质疏松患者骨密度(bone mineral density,BMD)的相关性。方法纳入2型糖尿病患者88例,根据骨密度T值分为3组:骨量正常组(T值>-1.0)(N组)、骨量减少组(-2.5<T值≤-1.0)(R组)和骨质疏松组(T值≤-2.5)...目的探讨Preptin在2型糖尿病性骨质疏松患者骨密度(bone mineral density,BMD)的相关性。方法纳入2型糖尿病患者88例,根据骨密度T值分为3组:骨量正常组(T值>-1.0)(N组)、骨量减少组(-2.5<T值≤-1.0)(R组)和骨质疏松组(T值≤-2.5)(OP组),比较3组患者一般临床资料、临床检验指标、骨密度、血浆Preptin以及结缔组织生长因子(connective tissue growth factor,CTGF)、骨特异性碱性磷酸酶(bone specific alkaline phosphatase,BAP)水平,利用Pearson相关分析研究BMD与一般资料、临床检验资料及骨代谢相关指标间的相关性,并采用SPSS 19.0统计学软件进行Preptin与CTGF、BAP相关曲线的绘制。结果3组患者体重、体质量指数差异有统计学意义(P<0.05)。3组患者骨密度以及骨代谢相关指标均存在显著差异,随着骨密度的降低,3组间Preptin、CTGF、BAP浓度逐渐降低,差异均有统计学意义(P<0.05)。L2~L4 BMD与Preptin、ALP-B、CTGF均呈正相关,Preptin与ALP-B、CTGF均呈正相关(P<0.05)。结论Preptin通过调节CTGF、BAP水平,在2型糖尿病性骨质疏松症的发生发展中发挥重要作用。展开更多
背景:中医药可有效防治糖皮质激素性骨质疏松,但其防治机制尚不明确。DKK1是Wnt/β-catenin信号通路的抑制剂,糖皮质激素可诱导其上调。因此,DKK1蛋白是防治糖皮质激素性骨质疏松的重要靶点。目的:探讨左归丸防治糖皮质激素性骨质疏松中...背景:中医药可有效防治糖皮质激素性骨质疏松,但其防治机制尚不明确。DKK1是Wnt/β-catenin信号通路的抑制剂,糖皮质激素可诱导其上调。因此,DKK1蛋白是防治糖皮质激素性骨质疏松的重要靶点。目的:探讨左归丸防治糖皮质激素性骨质疏松中对DKK1的调控作用。方法:18只3月龄雌性SD大鼠随机均分为3组:空白组、模型组和左归丸组。模型组和左归丸组大鼠皮下注射地塞米松建立糖皮质激素性骨质疏松模型;空白组给予等体积生理盐水;左归丸组皮下注射地塞米松同时给予左归丸水提液灌胃。1个月后取大鼠腰椎进行micro-CT检测骨量及骨微细结构、压缩试验检测生物力学性能,q PCR测定腰椎DKK1、Runx2和CTSK m RNA表达;血清检测碱性磷酸酶活性。结果与结论:(1)与空白组比较,模型组体积骨密度、相对骨体积、骨小梁数量和骨小梁厚度显著降低(P<0.05),骨小梁间距和结构模型指数较空白组显著增大(P<0.05),血清碱性磷酸酶活性较空白组呈降低趋势,DKK1 m RNA表达显著上调(P<0.05),成骨相关因子Runx2 m RNA表达呈下调趋势,破骨相关因子CTSK m RNA呈上调趋势;(2)与模型组比较,左归丸组体积骨密度、相对骨体积和骨小梁数量显著提升(P<0.05),结构模型指数显著降低(P<0.05),骨小梁间距有减小趋势但差异无显著性意义,血清碱性磷酸酶活性较模型组呈升高趋势,DKK1 m RNA表达显著下调(P<0.05),Runx2 m RNA表达呈上调趋势,CTSK m RNA呈下调趋势;(3)与模型组比较,左归丸组椎体压缩强度显著提升(P<0.05);(4)结果说明,左归丸可能通过下调DKK1的表达防治糖皮质激素性骨质疏松。展开更多
猪肌肉组织中碱性磷酸酶经预冷的正丁醇提取、硫酸铵分级沉淀、DEAE-52纤维素离子交换柱层析、Sephacryl S-200凝胶过滤层析得到碱性磷酸酶。纯化倍数为57.34,比活力为28.77U/mg。通过SDS-聚丙烯酰胺凝胶电泳(polyacrylamide gel electr...猪肌肉组织中碱性磷酸酶经预冷的正丁醇提取、硫酸铵分级沉淀、DEAE-52纤维素离子交换柱层析、Sephacryl S-200凝胶过滤层析得到碱性磷酸酶。纯化倍数为57.34,比活力为28.77U/mg。通过SDS-聚丙烯酰胺凝胶电泳(polyacrylamide gel electrophoresis,PAGE)分析可得该酶的分子质量为66kDa。该酶催化底物对硝基苯磷酸二钠(p-NPP)的最适pH值为9.7,最适温度为30℃,Mg2+和Ca2+对其有激活作用,Zn2+和EDTA对其有抑制作用。展开更多
文摘Hypophosphatasia (HPP) is an inherited skeletal disease caused by mutation of the gene encoding tissue non-specific alkaline phosphatase (TNSALP). Odonto-HPP is well known as the mildest of HPP. The manifestations involve only the teeth, such as premature primary teeth exfoliation caused by reduction of alveolar bone, enlarged dental pulp chamber, and dental defects. We report a case of a 9-years-old boy who developed HPP. He was observed from the primary dentition to the mixed dentition period. At initial presentation at our hospital, he had multiple premature exfoliation of primary teeth and reduction of the alveolar bone. HPP was suspected due to the low level of ALP activity in serum, his oral manifestation, and dental history. He was referred to a physician for the final diagnosis. Therefore his compound heterozygote mutations, c.1559 delT (T/delT) and c.407G > A (G/A), were found in TNSALP and he diagnosed with odonto-HPP. Even though these mutations were reported as being involved in odonto-HPP, his mineral densities tended to be lower than that of his age. It is therefore necessary to investigate the bone mineralization level in odonto-HPP without other bone symptoms. Moreover, ongoing enzyme-replacement therapy in odonto-HPP might improve dental abnormality and bone disorders.
文摘Background: Alkaline phosphatase has 4 isozymes, tissue non-specific alkaline phosphatase (TNAP), placental alkaline phosphatase (PLAP), intestinal alkaline phosphatase and germ-cell alkaline phosphatase. Hypophosphatasia (HPP) is an inherited skeletal disease caused by mutations of the gene encoding TNAP. Although TNAP is expressed in various tissues, the primary HPP symptoms appear in bones and teeth. The clinical severity of HPP varies widely from the most severe (perinatal, infantile and childhood) to the mildest forms (adult, and odonto-hypophosphatasia). We reported that gene therapy using a single injection of lentiviral vector expressing bone-targeted TNAP (TNAP-D10) is effective in preventing all the skeletal of HPP in TNAP knockout (Alpl–/–) mice as the model of infantile HPP. Objective: In this study we focus on evaluating the efficacy of treatment with gene therapy on the bone and teeth using TNAP-D10 and also we investigate the feasibility of gene therapy using bone-targeted PLAP (PLAP-D10). Methods and Findings: We used Alpl–/–mice that develop skeletal disease at postnatal days 6-8 mimicking the infantile form of human HPP. We injected 100 μl of lentiviral vectors harboring TNALP-D10 (5.0 × 107 TU) or PLAP-D10 (5.0 × 107 TU) to 1-day-old Alpl–/–?mice via the jugular vein. We performed histological analysis and micro-CT evaluation on bone and mandible of Alpl–/–?mice. The alveolar bone, enamel and dentin defects were corrected on treated Alpl–/–?mice by this treatment. Additionally the long bone growth rates (LGR) of long bones were encouraged on treated Alpl–/–?mice compared with untreated mice. Conclusions: These results indicate that the bone-targeted TNAP treatment mediated by lentivirus can correct not only the bone disorder but also the dental symptoms in Alpl–/–. This study also shows that PLAP-D10 can potentially be used to correct HPP disease.
文摘背景:中医药可有效防治糖皮质激素性骨质疏松,但其防治机制尚不明确。DKK1是Wnt/β-catenin信号通路的抑制剂,糖皮质激素可诱导其上调。因此,DKK1蛋白是防治糖皮质激素性骨质疏松的重要靶点。目的:探讨左归丸防治糖皮质激素性骨质疏松中对DKK1的调控作用。方法:18只3月龄雌性SD大鼠随机均分为3组:空白组、模型组和左归丸组。模型组和左归丸组大鼠皮下注射地塞米松建立糖皮质激素性骨质疏松模型;空白组给予等体积生理盐水;左归丸组皮下注射地塞米松同时给予左归丸水提液灌胃。1个月后取大鼠腰椎进行micro-CT检测骨量及骨微细结构、压缩试验检测生物力学性能,q PCR测定腰椎DKK1、Runx2和CTSK m RNA表达;血清检测碱性磷酸酶活性。结果与结论:(1)与空白组比较,模型组体积骨密度、相对骨体积、骨小梁数量和骨小梁厚度显著降低(P<0.05),骨小梁间距和结构模型指数较空白组显著增大(P<0.05),血清碱性磷酸酶活性较空白组呈降低趋势,DKK1 m RNA表达显著上调(P<0.05),成骨相关因子Runx2 m RNA表达呈下调趋势,破骨相关因子CTSK m RNA呈上调趋势;(2)与模型组比较,左归丸组体积骨密度、相对骨体积和骨小梁数量显著提升(P<0.05),结构模型指数显著降低(P<0.05),骨小梁间距有减小趋势但差异无显著性意义,血清碱性磷酸酶活性较模型组呈升高趋势,DKK1 m RNA表达显著下调(P<0.05),Runx2 m RNA表达呈上调趋势,CTSK m RNA呈下调趋势;(3)与模型组比较,左归丸组椎体压缩强度显著提升(P<0.05);(4)结果说明,左归丸可能通过下调DKK1的表达防治糖皮质激素性骨质疏松。
文摘猪肌肉组织中碱性磷酸酶经预冷的正丁醇提取、硫酸铵分级沉淀、DEAE-52纤维素离子交换柱层析、Sephacryl S-200凝胶过滤层析得到碱性磷酸酶。纯化倍数为57.34,比活力为28.77U/mg。通过SDS-聚丙烯酰胺凝胶电泳(polyacrylamide gel electrophoresis,PAGE)分析可得该酶的分子质量为66kDa。该酶催化底物对硝基苯磷酸二钠(p-NPP)的最适pH值为9.7,最适温度为30℃,Mg2+和Ca2+对其有激活作用,Zn2+和EDTA对其有抑制作用。
