用全血干化学和免疫浓缩技术测定骨碱性磷酸酶

骨碱性磷酸酶（ＢＡＬＰ）是由成骨细胞合成的，当小儿体内维生素缺乏时，骨钙化不足，成骨细胞活跃，ＢＡＬＰ上升，其改变先于影像学变化，被认为是诊断小儿佝偻病最特异、最敏感的指标。采用干化学和免疫浓缩技术的原理设计的骨碱性磷酸酶试剂盒，只需３０μｌ全血即可半定量读出骨碱性磷酸酶浓度。测定范围为１７０￣４２０Ｕ／Ｌ，其目测结果与ＷＧＡ亲和沉淀法的符合率为９２．２％。肝、胆和肠碱性磷酸酶对目测小儿血浆骨碱性     

辛伐他汀在人骨髓基质干细胞向成骨细胞分化过程中的作用

目的观察辛伐他汀对体外培养的人骨髓基质干细胞（Human Bone Marrow Stromal cells,hBMCs）成骨分化功能的影响,探讨其刺激成骨的作用机制。方法体外培养来自于外伤所致股骨颈骨折患者的骨髓基质干细胞,传代后实验组加入1×10^-7mol/L的辛伐他汀,在不同时间点采用ELISA检测核转录因子1（Core Binding Factor1,Cbfa1）与DNA的结合活性,碱性磷酸酶（Alkaline Phosphatase,ALP）比活性,及放射免疫法检测骨钙素（Osteocalcin,OCN）含量。结果在辛伐他汀作用后,实验组与对照组比较,实验组Cbfa1因子与DNA的结合活性增高,ALP比活性增高且骨钙素含量增加。结论1×10^-7mol/L辛伐他汀能够促进人骨髓基质干细胞成骨分化,此种促进作用可能与辛伐他汀增加其分化过程中相关转录因子的活性有关。     

续断骨伤合剂在促进骨折愈合中对血生化指标的影响

目的 观察续断骨伤合剂在促进骨折愈合中对血生化指标方面的影响.方法 将64只SD大鼠分为模型组和治疗组.均造成右胫骨骨缺损模型后分笼饲养,治疗组用续断骨伤合剂浓缩液灌胃,术后4d、7d、14d、28d取血样本和骨折标本.行CR撮片和血生化指标检测.结果 治疗组的血清ALP在第7、14、28大升高显著均高于对照组(P<0...     

骨质疏松患者血清FSH、E2、T、ALP水平的观察

目的:观察骨质疏松女性患者血清FSH 、E2、 T、ALP的含量.方法:根据临床诊断将81名妇女分为骨质疏松组和非骨质疏松组,分别检测血清FSH 、E2、 T、ALP水平.结果:骨质疏松的女性血清E2、 T水平显著低于非骨质疏松组(P＜0.01),血清FSH、ALP水平显著高于非骨质疏松组(P＜0.01).结论:中年后妇女如发现雌激素水平明显下降,应积极防治骨质疏松.     

辛伐他汀对正畸牙齿保持阶段骨形成的影响

目的:探讨全身给予辛伐他汀对大鼠正畸牙齿移动后保持阶段骨形成生化指标和骨密度的影响。方法:选用40只雄性Wistar大鼠,随机分成5组:基本对照组、阴性对照组(生理盐水)、低剂量组(2.5mg/kg)、中剂量组(5.0mg/kg)、高剂量组(10.0mg/kg),牵引其上颌第一磨牙向近中移动。实验组在加力装置去除前1d开始,腹膜下注射辛伐他汀;阴性对照组注射生理盐水,1次/d。4周后处死大鼠,取血,检测血清骨钙素(BGP)、碱性磷酸酶(ALP)、钙(Ca)、磷(P),测定上颌第一磨牙区牙槽骨密度(BMD)等。结果:实验组BGP、ALP水平较阴性对照组明显升高(P<0.05),低剂量组升高最明显,随剂量增加,BGP、ALP水平减低;实验组与对照组血清Ca、P无明显差别(P>0.05)。上颌第一磨牙区牙槽骨BMD对照组明显高于其它各组(P<0.05);实验组与阴性对照组比较,低剂量组最高,差异有显著性(P<0.05)。结论:辛伐他汀全身给药能够有效地促进局部牙槽骨骨合成代谢,最适有效剂量为2.5mg/kg。     

Multiple proliferation signaling pathways are modulated by octacalcium phosphate in osteoblasts

Octacalcium phosphate (OCP), a type of bioactive ceramics, may be associated with dentine, tooth apatite, and especially bone generation, and promotes wound healing after fracture. Recently, commercial bone grafting products containing a large amount of OCP material have been released because OCP can be synthesized in large quantities. It is reported to increase cell proliferation, but the interaction between OCP and cell signaling pathways is still unclear. In this study, first, we demonstrated OCP mediated cell signaling pathways with only purified OCP materials. OCP regulated P38, JNK (c-Jun N-terminal kinase), Src, and AKT (protein kinase B) signaling pathways. OCP crystals appeared in the characteristic ribbon shape but varied by several tens of micrometers in size. The X-ray diffraction pattern was the same as previously reported. We studied two concentrations of OCP (10 mg/ml and 20 mg/ml) to understand whether the effect of OCP on cell signaling pathways is dose dependent. We confirmed that OCP treatment affected cell proliferation and alkaline phosphatase and disrupted Src phosphorylation but did not change the total protein level. P38 phosphorylation was activated with OCP treatment and inhibited by SB203580, but P38 total protein level did not change. OCP inhibited JNK phosphorylation signaling, whereas PD98509 inhibited JNK phosphorylation with or without OCP. Interestingly, the AKT total level decreased after OCP treatment, but AKT phosphorylation increased considerably. Our results demonstrate that OCP materials modulate cell signaling pathways and increase cell proliferation.     

Clonogenicity of normal and malignant hematopoietic progenitor cells after exposure to synthetic alkyl-lymphospholipids

Summary Alkyl-lysophospholipids (ALP) are synthetic analogues of natural lysophosphatidylcholine and represent a new class of anti-tumor agents. They are cytotoxic in vitro with a high selectivity for neoplastic cells which, in contrast to normal cells, lack an alkyl-cleavage enzyme to degrade the adsorbed ALP molecules. As ALP accumulates, it interferes with normal membrane phospholipid turnover and eventually causes disruption of membrane integrity. To evaluate the potential value of ALP in eliminating leukemic cells from remission marrows prior to autologous transplantation, we tested the effect of various ALPs on the clongenicity of normal human marrow cells and on promyelocytic leukemia HL-60. A remarkable difference in the dose response to ALP of normal marrow cells an HL-60 was observed. After an incubation period of 24 h, the same inhibition of clonogenicity in HL-60 occurred at ALP concentrations 4 times lower than in normal marrow cells. Reducing the exposure time to 6 h enhanced the selectivity further: whereas HL-60 colonies were nearly completely inhibited at 16 g ALP/ml, more than 50% of normal CFU-c and BFU-E were recovered after incubation with 48 g/ml. No further increase in selectivity was achieved by changing the incubation temperature. Both thioether- and alkyl-analogues were active and no difference was observed between methoxy- and acylamino-substituted ALPs. We conclude that this selective cytotoxicity makes ALP compounds worth further study as purging agents in autologous bone marrow transplantation programs.Supported by the Deutsche Forschungsgemeinschaft An 111/3     

Osteoinductive capacity and heat stability of recombinant human bone morphogenetic protein-2 produced by <Emphasis Type="Italic">Escherichia coli</Emphasis> and dimerized by biochemical processing

One problem associated with clinical application of CHO-derived recombinant human bone morphogenetic protein (C-BMP-2) is its high cost due to the need for use of high doses. To solve this problem, Escherichia coli-derived BMP-2 (E-BMP-2) has been examined using the technique of molecular unfolding and refolding. However, it is unclear whether the characteristics of E-BMP-2 are appropriate for clinical application. In this study, we examined the biological activity of E-BMP-2 and its heat tolerance in in vitro and in vivo systems. SDS-polyacrylamide gel electrophoresis (SDS-PAGE) confirmed the high purity of E-BMP-2. E-BMP-2-induced alkaline phosphatase expression in osteoprogenitor cells (C2C12, ST2, and primary murine calvarial osteoblast cells) was dose-dependent, and consistently elicited ectopic new ossicles of significant size in mice, also in dose-dependent fashion. In addition, E-BMP-2 induced phosphorylation of Smad1/5/8 and mRNA expression of osteoblastic differentiation markers to the same extent as C-BMP-2. On the other hand, when E-BMP-2 was exposed to increasing heat over time, its bone-inducing capacity was maintained until reaching 70°C for 2 h or 90°C for 15 min. Thus, E-BMP-2 will exhibit a decrease in activity with the sterilization procedures required prior to use in surgery. These findings indicate that the biological capacity and heat stability of E-BMP-2 are almost equivalent to those of currently available C-BMP-2, and suggest that E-BMP-2 might, thus, solve current problems of cost impeding routine clinical use of rhBMP-2.     

Semichronic oral toxicity of cadmium: 1. Studies on rats

Cadmium (in the form of CdCl₂) was fed to groups of 20 male and 20 female rats each over a period of 3 months in concentrations of 0, 1, 3, 10 and 30 ppm. Appearance, behaviour, food consumption, growth and mortality of the treated rats of all groups were not affected during the 3-month period. The cadmium concentrations did not cause blood, liver or kidney damage. The systolic blood pressure of the treated animals was not increased. Autopsies and histopathological investigation of the animals showed no sign of any alterations. Cadmium accumulated dose-dependently in the kidneys and liver. Concentrations of cadmium up to 30 ppm in their feed were tolerated by rats over a period of 3 months without harm.