可吸收性医用珊瑚人工骨的致突变性研究

目的评价珊瑚人工骨的遗传毒性。方法采用Ames试验;细胞染色体畸变试验和小鼠骨髓细胞微核试验。结果不同浓度的浸提液加与不加S9mix条件下Ames试验;细胞染色体畸变试验以及微核试验与阴性对照组比较无显著差异,结果为阴性。结论在本试验系统条件下,可吸收性珊瑚人工骨无致突变作用。     

珊瑚型人工髋关节置换术的临床随访及松动原因的分析

自１９８３年至今应用珊瑚型人工髋关节置换术共５４例６０髋。其中人工全髋关节置换术３４例４０髋，人工股骨头置换术２０例２０髋。在５４例６０髋珊瑚型人工髋关节置换术中，有３７髋随访６个月～１３年，平均随访时间约７年，疗效满意率为８５．１％，其中假体松动４例占１４．８％，髋臼松动２例７．４％。假体松动原因是假体与股骨负重界面之间没有达到稳定接触，假体与髓腔形状不相匹配，修整髓腔松质骨时，髓腔扩大器应比假体小一号，避免假体与界面存留缝隙。对于髋臼发育不良的患者应避免髋关节旋转中心向外侧移位，应向内上方加深髋臼以减少水平移位距离，其次髋臼植骨加盖勿在负重区。     

自体骨髓基质干细胞复合珊瑚修复犬下颌骨节段缺损的初步研究

目的应用自体骨髓基质干细胞(bonemarrowstromalcells,BMSCs)复合珊瑚构建组织工程化骨,修复犬下颌骨节段性缺损。方法体外扩增培养、成骨诱导犬BMSCs。将第二代细胞复合珊瑚后修复犬自体右侧3cm的下颌骨节段缺损(n=6);以单纯珊瑚植入缺损处为对照(n=6),术后12、32周分别通过影像学,大体形态观察,组织学和生物力学的方法检测骨缺损的修复效果。结果成骨诱导的BMSCs在珊瑚支架上生长良好。X线片显示12周时实验组骨痂较多,对照组材料明显吸收;32周时CT、X线片和大体观察显示术后实验组骨愈合良好,对照组为骨不连;骨密度检测示实验组显著高于对照组(P<0.05);组织学示实验组有较多成熟骨呈骨性愈合,对照组为纤维性愈合;生物力学测试实验组与正常下颌骨力学强度差异无统计学意义(P>0.05)。结论自体成骨诱导BMSCs复合珊瑚形成的组织工程化骨可修复犬下颌骨节段缺损。     

珊瑚涂层人工骨的制备及溶血实验研究

目的根据生物工程及化工等原理,制备珊瑚人工骨,并检测此珊瑚(Coral)复合人工骨的血液相容性。方法将制备的珊瑚复合人工骨、按照国际标准化组织和我国国家标准规定要求,采用将珊瑚复合人工骨及其浸提液与抗凝稀释兔血直接接触,测定红细胞释放的血红蛋白量(OD值),将测得的OD值换算为溶血率。结果珊瑚复合人工骨材料浸提液组的溶血率为1．81％,材料与血液直接接触组溶血率为1．41％,阴性对照组和阳性对照组溶血率分别为0％、100％。结论珊瑚复合人工骨材料无溶血作用,血液相容性良好。     

Pharmacological and structural characterization of a novel phospholipase A2 from Micrurus dumerilii carinicauda venom.

We have isolated a new phospholipase A2 (MiDCA1) from the venom of the coral snake Micrurus dumerilii carinicauda. This toxin, which had a molecular mass of 15,552Da, shared high sequence homology with the PLA2 toxins MICNI A and B from Micrurus nigrocinctus venom (77.7% and 73.1%, respectively). In chick biventer cervicis preparations, MiDCA1 produced concentration- and time-dependent neuromuscular blockade that reached 100% after 120 min (2.4 microM, n = 6); contractures to exogenously applied carbachol (8 microM) and KCl (13 mM) were still seen after complete blockade. In mouse phrenic-nerve diaphragm preparations, MiDCA1 (2.4 microM; n = 6) caused triphasic changes followed by partial neuromuscular blockade. Intracellular recordings of end-plate potentials (EPPs) and miniature end-plate potentials (MEPPs) from mouse diaphragm preparations showed that MiDCA1 increased the quantal content by 386+/-12% after 10 min (n = 14; p<0.05) and caused a triphasic change in the frequency of MEPPs. MiDCA1 also decreased the resting membrane potential, an effect that was prevented by tetrodotoxin and/or low extracellular calcium, but not by d-tubocurarine. The toxin increased the amplitude of mouse sciatic-nerve compound action potentials by 30+/-9% (0.6 microM; p<0.05). Potassium currents elicited in freshly dissociated dorsal root ganglia neurones were blocked by 31+/-1% (n = 4; p<0.05) in the presence of 2.4 microM MiDCA1. These results show that MiDCA1 is a new presynaptic phospholipase A2 that produces neuromuscular blockade in vertebrate nerve-muscle preparations. The triphasic effects seen in mammalian preparations and the facilitatory response were probably caused mainly by the activation of sodium channels, complemented by the blockade of nerve terminal potassium channels. The inability of d-turocurarine to prevent the depolarization by MiDCA1 indicated that cholinergic nicotinic receptors were not involved in this phenomenon.     

Functional characterization of Vitellogenin_N domain,domain of unknown function 1943, and von Willebrand factor type D domain in vitellogenin of the non-bilaterian coral Euphyllia ancora: Implications for emergence of immune activity of vitellogenin in basal metazoan

Our understanding of the function of vitellogenin (Vg) in reproduction has undergone a transformation over the past decade in parallel with new insights into the role of Vg in immunity. However, the time when Vg was endowed with immunological activities during animal evolution remains elusive. Here we demonstrate for the first time that the recombinant proteins rVitellogenin_N, rDUF1943, and rVWD from Vg of the basal metazoan coral Euphyllia ancora not only interact with Gram-positive and negative bacteria as well as their conserved surface components LTA and LPS but also enhance phagocytosis of bacteria by macrophages. Moreover, challenge with LPS results in a marked up-regulation of vg in the coral E. ancora. These data suggest that E. ancora Vg, like that described in the bilaterian oviparous animals fish and amphioxus, is a molecule related to antibacterial defense, indicating that the timing of the emergence of immune role of Vg predates the divergence of the cnidarian (non-bilaterian) and bilaterian lineages.     

In Vitro Regulation of CaCO3 Crystal Growth by the Highly Acidic Proteins of Calcitic Sclerites in Soft Coral,Sinularia Polydactyla

Acidic proteins are generally thought to control mineral formation and growth in biocalcification. Analysis of proteinaceous components in the soluble and insoluble matrix fractions of sclerites in Sinularia polydactyla indicates that aspartic acid composes about 60% of the insoluble and 29% of the soluble matrix fractions. We previously analyzed aspartic acids in the matrix fractions (insoluble = 17 mol%; soluble = 38 mol%) of sclerites from a different type of soft coral, Lobophytum crassum, which showed comparatively lower aspartic acid-rich proteins than S. polydactyla. Thus, characterization of highly acidic proteins in the organic matrix of present species is an important first step toward linking function to individual proteins in soft coral. Here, we show that aspartic-acid rich proteins can control the CaCO₃ polymorph in vitro. The CaCO₃ precipitates in vitro in the presence of aspartic acid-rich proteins and 50 mM Mg²⁺ was verified by Raman microprobe analysis. The matrix proteins of sclerites demonstrated that the aspartic-acid rich domain is crucial for the calcite precipitation in soft corals. The crystalline form of CaCO₃ in the presence of aspartic acid-rich proteins in vitro was identified by X-ray diffraction and, revealed calcitic polymorphisms with a strong (104) reflection. The structure of soft coral organic matrices containing aspartate-rich proteins and polysaccharides was assessed by Fourier transform infrared spectroscopy. These results strongly suggest that the aspartic acid-rich proteins within the organic matrix of soft corals play a key role in biomineralization regulation.     

A Comparative Study on Morphochemical Properties and Osteogenic Cell Differentiation within Bone Graft and Coral Graft Culture Systems

The objective of this study was to compare the morphological and chemical composition of bone graft (BG) and coral graft (CG) as well as their osteogenic differentiation potential using rabbit mesenchymal stem cells (rMSCs) in vitro. SEM analysis of BG and CG revealed that the pores in these grafts were interconnected, and their micro-CT confirmed pore sizes in the range of 107-315 µm and 103-514 µm with a total porosity of 92% and 94%, respectively. EDS analysis indicated that the level of calcium in CG was relatively higher than that in BG. FTIR of BG and CG confirmed the presence of functional groups corresponding to carbonyl, aromatic, alkyl, and alkane groups. XRD results revealed that the phase content of the inorganic layer comprised highly crystalline form of calcium carbonate and carbon. Atomic force microscopy analysis showed CG had better surface roughness compared to BG. In addition, significantly higher levels of osteogenic differentiation markers, namely, alkaline phosphatase (ALP), Osteocalcin (OC) levels, and Osteonectin and Runx2, Integrin gene expression were detected in the CG cultures, when compared with those in the BG cultures. In conclusion, our results demonstrate that the osteogenic differentiation of rMSCs is relatively superior in coral graft than in bone graft culture system.     

二十五味珊瑚丸联合西比灵对偏头痛患者血浆内皮素的影响及疗效观察

目的观察二十五味珊瑚丸联合西比灵治疗偏头痛的疗效和不良反应。方法将158例偏头痛患者随机分为两组,观察组(84例)给予二十五味珊瑚丸联合西比灵治疗,对照组(74例)单用西比灵治疗,两组疗程均为4周。对两组患者治疗前后头痛的改善情况进行疗效评定,并用汉密尔顿抑郁量表(HAMD)和汉密尔顿焦虑量表(HAMA)对患者进行评分。采用放射免疫分析技术测定血浆内皮素含量。结果与单用西比灵治疗的对照组比较,二十五味珊瑚丸和西比灵联合治疗的观察组临床疗效好,总有效率高,可以更有效地减轻头痛的程度,缓解伴随的非头痛症状如焦虑、抑郁,不仅如此,联合治疗的观察组中血浆内皮素水平在治疗后下降更明显。在治疗过程中两组均未出现明显不良反应。结论二十五味珊瑚丸和西比灵联合治疗偏头痛临床疗效好,值得推广使用。     

Dietary coral calcium and zeolite protects bone in a mouse model for postmenopausal bone loss

In patients diagnosed with osteoporosis, calcium is lost from bones making them weaker and easily susceptible to fractures. Supplementation of calcium is highly recommended for such conditions. However, the source of calcium plays an important role in the amount of calcium that is assimilated into bone. We hypothesize that naturally occurring coral calcium and zeolite may prevent ovariectomy-induced bone loss. We have measured bone loss in ovariectomized mice supplemented with coral calcium and Zeolite. Female C57BL/6 mice were either sham-operated or ovariectomized and fed diets containing coral calcium or zeolite for 6 months. Serum was analyzed for bone biochemical markers and cytokines. Bones were analyzed using dual x-ray absorbtiometry, peripheral quantitative computed tomography, and micro–computed tomography densitometry. In the distal femoral metaphysis, total bone and cortical bone mass was restored and the endocortical surface was significantly decreased in coral calcium and zeolite fed ovariectomized (OVX) mice. Trabecular number and the ratio of bone volume to total volume was higher in OVX mice after coral calcium and zeolite feeding, while trabecular separation decreased in the different treatment OVX groups. Coral calcium protected bone to a lesser extent in the proximal tibia and lumbar vertebrae. Overall, coral calcium and zeolite may protect postmenopausal bone loss.