藻酸钙凝胶-成骨细胞-骨粉构建工程化骨修复兔颅骨缺损的实验研究

目的探讨藻酸钙凝胶、成骨细胞、骨粉复合构建的可塑形组织工程骨修补兔颅骨缺损后的形态学变化和成骨效果.方法28只日本大耳白兔,随机分为A(n=20)、B(n=8)两组,手术在兔颅顶骨矢状缝两侧分别各建立一个直径1cm的圆形全层缺损,用两种方法藻酸钙凝胶、成骨细胞、骨粉和藻酸钙凝胶、骨粉分别构建组成可塑形的组织工程骨复合材料,分别填补修复A组兔颅骨左右两侧的缺损,B组为空白对照组,通过大体、组织学、X线观察材料的形态变化、成骨情况,并对X线片和组织学切片进行评分.结果材料植入后,局部未见红肿、积液、渗出等异常反应.①藻酸钙凝胶-成骨细胞-骨粉组修补后12周颅骨缺损基本被硬性组织所修复,镜下见修复材料大多被骨组织替代,骨粉基本被吸收,有块状凝胶残留其中,组织学评分为(5.50±1.00).X线片见兔颅骨缺损处有高密度骨痂影存在,布满整个缺损区,X线片评分为(3.25±0.95).②藻酸钙凝胶-骨粉组修补后12周部分颅骨缺损被硬性组织所修复,镜下见修复材料部分转变成骨组织,骨粉基本被吸收,有凝胶残留其中,组织学评分为(3.25±1.50).X线片见兔颅骨缺损处有高密度骨痂影存在,主要分布在缺损区的边缘部位,X线片评分(2.25±0.25).③空白对照组骨缺损主要被膜样纤维组织修复,在紧邻骨缺损边缘处有硬性组织形成,镜下见修复组织边缘有骨组织存在,中央大部为膜状致密纤维组织,组织学评分为(1.50±0.50),X线片仅见靠近骨缺损边缘的部位存在致密骨痂影,X线片评分为(1.00±0.57).结论藻酸钙凝胶、成骨细胞、骨粉构建的可塑形组织工程骨可根据颅骨缺损的形态进行塑形填补,在体内有良好的成骨能力,可达到对兔颅骨缺损的骨性修复,但部分藻酸盐凝胶吸收缓慢,手术后12周仍不能满意吸收.     

TAT和GMP-140在急性心肌梗塞患者溶栓治疗中的应用价值

目的探讨急性心肌梗塞溶栓治疗中凝血酶-抗凝血酶复合物(TAT)和血小板α-颗粒膜蛋白(GMP-140)动态变化及临床应用价值.方法以健康人为正常对照组(n=48),随机选择6h内急性心肌梗塞患者48例,用150万U尿激酶进行溶栓治疗,并按临床再通标准分为再通组(n=30),未通组(n=18),分别在溶栓前、溶栓后2h、4h、24h、48h采用酶联免疫吸附双抗体夹心法测定血浆TAT和GMP-140水平,观察其动态变化.结果患者组治疗前TAT水平(14.56±1.04)μg/L、GMP-140水平(12.50±1.05)ng/ml,较正常对照组(2.15±0.89)μg/L、(8.39±1.33)ng/ml显著升高(P<0.05),治疗后2h再通组GMP-140水平达到峰值(22.17±2.05)ng/ml,4h后TAT到达峰值(46.67±5.79)μg/L(P<0.05)后逐渐下降,持续至少3天,但仍高于正常对照组.未通组二项指标持续缓慢上升,48h达到峰值.结论急性心肌梗塞溶栓治疗前后TAT、GMP-140水平变化对判断疗效有一定指导意义.     

弓形虫真核表达质粒pcDNA3/GRA1的构建及序列测定

目的 :构建弓形虫致密颗粒抗原 1 (GRA1 )真核表达质粒 ,为进一步开展DNA疫苗的保护性研究打下基础。方法 :采用PCR扩增出编码GRA1目的基因 ,用EcoRⅠ /XhoⅠ分别对扩增产物和真核表达质粒pcDNA3进行双酶切 ,将GRA1定向克隆到pcDNA3EcoRⅠ /XhoⅠ位点 ;对重组质粒进行PCR ,双酶切初步鉴定后做序列测定。结果 :特异扩增出预计的GRA1片段 ,大小为 5 73bp ;扩增产物双酶切后成功连接到pcDNA3中 ,经PCR ,双酶切及序列测定结果表明重组质粒中含有GRA1读框。结论 :成功构建弓形虫GRA1真核表达质粒。     

风痛灵颗粒制备工艺研究

目的：为了优化风痛灵颗粒的制各工艺。方法：采用正交试验以阿魏酸、多糖等为指标。结果：得出最佳的浸提工艺参数，同时考察了最佳的喷雾干燥及制粒条件。结论：本方法是风痛灵颗粒的最佳的制备方法。     

Activity-dependent survival and enhanced turnover of calcium in cultured rat cerebellar granule neurons

Neurons survive when their activity is maintained. An influential hypothesis on the cellular mechanism underlying this phenomenon is that there is an appropriate range of intracellular Ca²⁺ concentration ([Ca²⁺]i) for survival. The rat cerebellar granule neuron in culture serves as the most often used model system for the analysis of activity-dependent survival, since it does not survive unless an excitant (KCl or glutamate) is added to the culture medium. Against the above-mentioned hypothesis, we found in our previous examination no difference between steady-state [Ca²⁺]i in granule neurons cultured under high KCl (i.e., survival) and low KCl (i.e., death) conditions. In this report, we present the quantitative background of unchanged [Ca²⁺]i between the two culture conditions. Influx of Ca²⁺ due predominantly to L-type voltage-dependent calcium channels was higher in high KCl cultures than in low KCl cultures. At the same time, efflux of Ca²⁺ due to the activity of Ca²⁺/Na⁺ antiport was also higher in high KCl cultures. Additionally, we found that the endocytotic activity was greater in high KCl cultures than in low KCl cultures, as monitored by the rate of uptake of horseradish peroxidase added to medium. Since the uptake was blocked by an internal Ca²⁺ chelator, the increased endocytotic activity in high KCl cultures might be a consequence of the enhanced Ca²⁺ turnover.     

Toxic effect of glutamate causes mitochondria damage in granule cells of dissociated cultures of rat cerebellum

It is shown that treating rat cerebellum with glutamate in a neurocytotoxic concentration causes a drop of the mitochondrial membrane potential in granule cells and leads to ultrastructural alterations of mitochondria in these neurons. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 119, N ^o 4, pp. 378–380, April, 1995. Presented by the late Member of the Russian Academy of Medical Sciences     

Engulfment of mast cell secretory granules on skin inflammation boosts dendritic cell migration and priming efficiency

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Dimebon and Tacrine Inhibit Neurotoxic Action of β-Amyloid in Culture and Block L-type Ca2+ Channels

Dimebon, a Russian-made drug, inhibited toxic effects of beta -amyloid on cultured neurons. Excessive accumulation of beta-amyloid in the brain is characteristic of Alzheimer dementias. Antialzheimer preparations tacrine and dimebon improve survival of cerebellar granule cells during long-term incubation with Ab25-35, the neurotoxic fragment of beta-amyloid. Both preparations can block potential-dependent Ca²⁺ entry into neurons by about 20%, which is explained by their selective action on L-type Ca²⁺ channels. It was assumed that the neuroprotective effect of dimebon and tacrine against Ab25-35 partially depends on inhibition of potential-dependent Ca²⁺ entry.