新型骨与软骨组织工程支架材料制备及其性能研究

目的：研究符合骨与软骨组织工程制备技术要求的新型支架复合材料；并确定其性能与结构优化设计的主要参数指标。方法：按不同CPPf(聚磷酸钙纤维）／PLL（L－聚乳酸）重量比及相同CPPfPLLA/NaCl颗粒重量比，采用溶铸颗粒沥取（Solventcasting particulte-leaching)法轩系列CPPf/PLLA支架复合材料；液体置换法，压缩性能测试及扫描电子显微镜观察其密度，孔隙度压缩模量及微结构特征；材料样品体外人工降解液直接浸渍法判定其生物降解性能，并观测其生物降解率，压缩碍及微结构由微孔海绵状过渡为纤维网状。随着支架材料在人工降解液中降解时间的延长，其降解率以0％－13．2％线性递增；压缩模量呈线性降低至1.10-2.78MP；材料横截面微结构则发生相应改变。结论 CPPf/PLLA支架复合材料具有高孔隙度的三维立体结构，良好的抗压缩性能和和解性能，经进一步优化设计后，可成为结构和性能符合各项要求的新与软骨组织工程支架材料。     

Cobalt-dependent potentiation of net inward current density in Helix aspersa neurons.

A low concentration of transition metal ions Co2+ and Ni2+ increases the inward current density in neurons from the land snail Helix aspersa. The currents were measured using a single electrode voltage-clamp/internal perfusion method under conditions in which the external Na+ was replaced by Tris+, the predominant external current carrying cation was Ca2+, and the internal perfusate contained 120 mM Cs+/0 K+; 30 mM tetraethylammonium (TEA) was added externally to block K+ current. In the presence of Co2+ (3 mM) or Ni2+ (0.5 mM) inward Ca2+ currents were stimulated normally by voltage-dependent activation of Ca2+ channels. There was a 5-10% decrease in the rate of rise of the inward current. The principal effect of Co2+ and Ni2+ in increasing the current density seems to be a decrease in the rate at which the inward currents decline during a depolarizing voltage pulse. The results may be due to a decrease in a voltage-dependent or Ca(2+)-dependent outward current and/or an inhibition of Ca2+ channel inactivation. Outward current under these conditions (zero internal K+) was significant and most likely due to Cs+ efflux through the voltage-activated or Ca(2+)-activated nonspecific cation channels. Co2+ is an extremely effective blocker of this outward current. These results are not an artifact of internal perfusion or the special ionic conditions. Intracellular recording of unperfused neurons in normal Helix Ringer's solution showed that the Ca(2+)-dependent action potential duration was increased significantly by low concentrations of Co2+. This result is consistant with the Co(2+)-dependent increase in inward (depolarizing) current seen in voltage-clamp experiments.     

Influence of a calcium dependent protease inhibitor on platelet activation and secretion

Continuous proteolysis resulting in consumption of major cytoskeletal proteins may be essential for platelet activation and aggregation. In this study we evaluated the effect of a known protease inhibitor, Leupeptin, on agonist induced platelet aggregation and secretion. Platelets exposed to 10 ugs/ml of Leupeptin did not aggregate in response to the action of thrombin (0.2u/ml). However, a concentration of Leupeptin as high as 250 ugs/ml did not prevent arachidonate induced aggregation and secretion. Leupeptin (100 ugs/ml) effectively blocked thrombin (0.2 u/ml) induced elevation of cytosolic calcium, but did not affect arachidonate induced elevation of platelet intracellular calcium levels. At a concentration of 100 ug/ml, Leupeptin effectively blocked thrombin (0.5u/ml) induced clot formation of platelet poor plasma, suggesting that it can exert its effect on thrombin by preventing fibrinogen degradation. Effective K_i for the competitive inhibition of thrombin induced hydrolysis of a chromogenic substrate, S2238, by Leupeptin was 2.4 uM. Leupeptin inhibition of platelet function was reversible by washing platelets free of the polypeptide. Results of our study demonstrate that Leupeptin inhibits thrombin induced platelet activation, probably by interfering with its proteolytic activity on the platelet surface membrane. However, inhibition of platelet surface membrane associated proteases did not prevent activation of platelets by other agonists.     

甲基黄酮醇胺盐酸盐对异丙肾上腺素正性频率作用的影响

本文比较了甲基黄酮醇胺盐酸盐(MFOA)、普萘洛尔(Pro)、维拉帕米(Ver)对异丙肾上腺素(Iso)所致兔离体右心房正性频率作用的影响。Pro使Iso累积浓度反应曲线平行右移,不抑制最大反应,属于典型的竞争性抑制剂,其pA2=8.43;MFOA和Ver使Iso量效曲线向下右移,抑制最大反应,为非竞争性拮抗。MFOA(2×10~(-5)M)和Ver(2×10~(-7)M)分别使最大反应下降19.28％和48.57％,其pD'2值分别为4.07±0.14、6.68±0.15。结果表明MFOA的作用不同于Pro,和Ver相似。     

Effect of intraperitoneally injected fluoride on plasma calcium in suckling and adult rats

Summary Sodium fluoride 10, 20, or 40 mg/kg body weight was given intraperitoneally to rats (6–11 days old and 90–95 days old). Blood analyses showed an initial increase in plasma fluoride concentration. The subsequent decrease in fluoride was paralleled by a decrease in total plasma calcium. These plasma concentrations were normal at blood collection 4 days after fluoride injection. The baby rats differed from the older rats in that their initial plasma calcium was higher and that the drop in plasma calcium concentration was less pronounced than in the old rats. A diet low in calcium and phosphate enhanced the effects of fluoride on total plasma calcium. The data indicate that the effect of large doses of fluoride on lowering the plasma calcium level is modified by the calcium intake.     

Distribution of cytochrome oxidase and parvalbumin in the primary visual cortex of the adult and neonate monkey,Callithrix jacchus

The anatomical distributions of the mitochondrial enzyme cytochrome oxidase (CO) and of the calcium binding protein parvalbumin (PV) were studied in the striate cortex of adult and neonate New World monkeys (Callithrix jacchus). In the adult marmoset, both proteins were found in laminar arrangements similar to those described for the macaque monkey, with prominent bands of PV-like immunoreactive (PV-LI) puncta in layers IV and IIIb, and fairly evenly distributed PV-LI nonpyramidal neurons. Furthermore, the pattern of CO activity in area 17 of the neonate marmoset was almost identical to the CO pattern described in neonate macaque and squirrel monkeys. It came, therefore, as a surprise to find that the adult pattern of PV-like immunoreactivity (PV-LI) in the marmoset striate cortex arises from a neonatal pattern strikingly different from that seen in any developmental stage of the macaque, or in any other mammal studied so far. In the deep layers IV through VI of the neonate marmoset, a large number of PV-LI neurons was stained in bandlike patterns, their number in layers IV and V exceeding the number of PV-LI neurons present in these layers of the adult marmoset area 17. Staining of layers IV and VI was restricted to area 17 and involved nonpyramidal cells and their exceeding the number of PV-LI neurons present in these layers of the adult marmoset area 17. Staining of layers IV and VI was restricted to area 17 and involved nonpyramidal cells and their processes. The stained band of layer V, in contrast, continued throughout most of the neocortex. In area 17, an estimated 10 to 20% of the stained cells in layer V exhibited pyramidal shapes. The findings show that the expression of PV by visual cortical cells occurs before birth and suggest that the comparatively early onset of PV expression is not dependent on the onset of textured vision. The exuberant number of stained cells in some layers, and particularly the staining of pyramidal cells, in the neonate marmoset, suggest that a considerable number of cells possesses the stainability for PV-LI only transiently, i.e., in the marmoset, these cells have a specific demand for parvalbumin during this phase of their development. © 1994 Wiley-Liss, Inc.     

钙和鸡冠花黄酮提取物对大鼠废用性骨质疏松的作用

目的探讨钙和鸡冠花黄酮提取物对废用性骨质疏松大鼠的作用,为骨质疏松的防治提供依据.方法选用健康SD大鼠40只,雄性.随机分为5组,第1、2组分别为正常对照和阳性对照,灌胃蒸馏水5 ml/(kg@d),第2～5组制造废用性骨质疏松大鼠模型,第3、4、5组为预防组,分别灌胃等量鸡冠花提取物,碳酸钙,鸡冠花提取物加钙溶液.实验8周后,测骨密度(BMD),股骨CT值,测血清MDA、SOD及24 h尿钙、羟脯氨酸(HOP)、尿肌酐含量等.结果阳性组与正常对照组比较股骨重、灰分重、CT值、血清钙下降,尿钙、尿HOP升高(P＜0.05或0.01),预防组与阳性组比较股骨重、CT值显著增加,碳酸钙组、鸡冠花加钙组灰分重显著增加(P＜0.05).与阳性组比较各预防组血清钙增加,尿钙、尿HOP减少(P＜0.05或＜0.01).鸡冠花组、鸡冠花加钙组与阳性组比较股骨BMD、血清SOD升高,MDA降低,单纯补钙组仅BMD升高(P＜0.05或＜0.01).结论鸡冠花提取物、钙对废用性骨质疏松大鼠骨质代谢均有促进作用,两者合用具有更好的预防效果,能延缓或减轻废用性骨质疏松的发生.     

Effects of Clodronate on Cortical and Trabecular Bone in Ovariectomized Rats on a Low Calcium Diet

The aim of this study was to evaluate the contribution of a low calcium diet to the cortical and trabecular osteoporosis seen in ovariectomized rats after 7 weeks on a low calcium diet and to investigate the effects of the bisphosphonate clodronate on this development of osteoporosis. Thirty-six mature, female Wistar rats were randomized into four groups: Ovx−B (bisphosphonate) and Ovx−C (control) were ovariectomized, and Sham−Ca (low calcium) and Sham+Ca (normal calcium) were sham operated. The first three groups were fed a low calcium diet (0.01%) and Sham+Ca normal rat chow (Ca 1.1%). The Ovx−B received 10 mg/kg s.c. clodronate daily for nine weeks, and Ovx−C, Sham−Ca, and Sham+Ca received the same volumes of saline. Bone mineral turnover measured as ⁸⁵Sr-uptake was increased in all low calcium groups compared to Sham+Ca. The Sham+Ca femora had higher dry weight and ash weight than the other groups, and Ovx−C had higher dry weight compared with Ovx−B and Sham−Ca. Calcium content was lower in both Ovx groups compared to both Sham groups. Magnesium was lower in all groups compared to Sham+Ca and higher in Ovx−B compared with Ovx−C. In the femoral shaft, Sham+Ca had significantly higher ultimate bending moment, energy absorption, and deflection compared to the other three groups. Ultimate bending moment was higher in Sham−Ca than in Ovx−C. Stiffness was increased in both Sham+Ca and Ovx−B compared to Ovx−C. The maximum stress in the femoral midshaft was higher in Sham+Ca than in the other groups, and higher in Ovx−B than in Ovx−C. Histomorphometry showed increased medullary area in all low calcium groups compared to Sham+Ca and larger cortical area in Sham+Ca and Ovx−B compared to Ovx−C. Compared to Sham+Ca the trabecular bone volume was decreased to 30% in Sham−Ca and to 9% in Ovx−C, but was unchanged in Ovx−B. The low calcium diet generally increased bone mineral turnover and reduced the tibial bone volume. Femoral changes led to a reduction of cortical fracture strength and maximal stress. Ovariectomy in addition to a low calcium diet reduced femoral strength even more. Daily injections of clodronate to ovariectomized rats on a low calcium diet increased femoral shaft stiffness and maximum stress, and clodronate preserved both trabecular and cortical tibial bone volume completely. Received: 11 June 1996 / Accepted: 5 March 1997     

一种中西药复方制剂(PESM)对大鼠实验性肾结石形成的抑制作用

用乙二醇法诱导大鼠产生草酸钙结石，用45~Ca示踪测定法了解中草药复方制剂（PESM）对草酸钙结石形成的抑制作用。结果显示，实验组肾脏放射性强度明显减少（实验组3.47±3.52cpm／mg，对照组10.89±14.24cpm／mg）。干肾重量明显下降（实验组306.0±47.0mg，对照组343.1±59.4mg），说明PESM能明显抑制肾脏草酸钙结石的形成。