Morphologic characterization of osteoblast-like cell cultures isolated from newborn rat calvaria

Summary Two methods for harvesting osteoblast-like cell populations from newborn (10 days) rat calvaria were compared. The first one consisted in culturing the periosteum-free bones and then trypsinizing the cells on the bone surface. The second one involved the migration of the osteoblasts on glass fragments before trypsinization. Since the plating efficiency, the proportion of alkaline phosphatase-positive cells, the population doubling time, and the calcium deposition were more adequate, the second method was used to further characterize the behavior of the cultures. During the first week of culture, the cells featured shapes similar to those observedin vivo on the surface of periosteum-free calvaria. They formed multilayers and, in the presence of ascorbic acid, synthetized an organic matrix containing exclusively type I collagen. Later, small amounts of type III collagen appeared. The cells were embedded in the matrix and progressively acquired the morphologic phenotype of osteocyte-like cells. The matrix mineralized in the presence of β-glycerophosphate. The technique of dropinoculation (high concentration of cells in a small volume of medium) promoted the multilayer formation and the achievement of large mineralized plates (about 1 cm²) in 3 weeks of culture.     

固生蛋白tenascin对肝细胞癌血管生成及浸润转移的影响

目的　探讨固生蛋白(tenascin ,TN)在肝细胞癌(hepatocellularcarcinoma ,HCC)中的表达和与HCC血管生成及浸润转移的关系。方法　应用免疫组织化学法检测4 2例HCC、10例肝硬化和7例正常肝组织内TN的表达情况,观察与HCC病理学特点及微血管密度(microvesseldensity ,MVD)之间的关系。结果　①TN在HCC中的阳性表达率和强度显著高于肝硬化和正常肝脏组织( χ2 =4 15 ,P <0 0 5 ;t=2 4 73,P <0 0 5 ) ;②TN在有包膜侵犯组、病理分级Ⅲ～Ⅳ组及有转移HCC中的阳性表达率及强度明显高于无包膜侵犯组( χ2 =5 4 7,P <0 0 5 ;t=2 138,P <0 0 5 )、病理分级Ⅰ～Ⅱ级组( χ2 =6 87,P <0 0 1;t=2 4 79,P <0 0 5 )以及无转移组( χ2 =10 6 ,P <0 0 1;t=2 6 93,P <0 0 1) ;③肝癌>5cm、有包膜侵犯、有转移、病理分级Ⅲ～Ⅳ级组的MVD值明显高于肝癌≤5cm(t=2 0 36 ,P <0 0 5 )、无包膜侵犯(t =2 32 8,P <0 0 5 )、无转移(t =2 4 94 ,P <0 0 5 )及病理分级Ⅰ～Ⅱ级组(t =2 2 71,P <0 0 5 )。④TN阳性表达HCC中的MVD值明显高于TN阴性表达HCC中的MVD值(t =2 4 87,P <0 0 5 )。结论　TN在HCC组织中的表达上调与其血管生成和浸润转移有关     

低密度及氧化低密度脂蛋白对内皮细胞分泌肾上腺髓质素的影响

目的探讨低密度及氧化低密度脂蛋白对内皮细胞分泌肾上腺髓质素(ADM)的影响.方法利用培养的内皮细胞株ECV-304细胞,分别与不同浓度的低密度(50 100mg/L)、氧化低密度脂蛋白(50 100mg/L)及低密度脂蛋白(50 mg/L) 氧化低密度脂蛋白(50 mg/L)进行孵育,24 h后分别收集培养上清及细胞,用放免分析检测上清及细胞内肾上腺髓质素的含量.结果低密度脂蛋白对肾上腺髓质素的分泌无影响,而氧化型低密度脂蛋白能明显刺激ADM的分泌,二者合用的作用接近于100mg/L的OX-LDL.结论OX-LDL可能具有氧化LDL使其成为OX-LDL的作用,ADM的分泌可能是对细胞损伤的一种反应.     

Relationship Among MRTA, DXA, and QUS

Dual-energy X-ray absorptiometry (DXA) and quantitative ultrasound (QUS) are the accepted modalities for the evaluation of fracture risk in the clinical setting. However, neither method provides a direct measurement of bone mechanics. In this study, we investigated a prototype device, known as a mechanical response tissue analyzer (MRTA), which provides direct mechanical measurements of mechanical properties of bone. A total of 56 healthy volunteers (20 men and 36 women) between the ages of 18 and 83 were recruited. The MRTA was used to measure the cross-sectional bending stiffness (EI) of the ulna bone. Axial speed of sound (SOS) at the ulna bone was determined by QUS; bone mineral content (BMC) and bone mineral density (BMD) were determined by DXA. Correlations, regression analysis, and analyses of variance (ANOVAs) were used to compare the three modalities. These analyses revealed that although there are strong linear relationships among the data collected by the various technologies, the bone properties reflected by MRTA are not fully explained by DXA and QUS. We conclude that the total information conveyed by MRTA measurements is unique. Further research is needed to delineate the different qualities of bone strength that are captured by MRTA, but not by DXA or QUS.     

Prevalence of silent myocardial ischemia in asymptomatic individuals with subclinical atherosclerosis detected by electron beam tomography

BACKGROUND: Electron beam tomography coronary calcium imaging is an evolving technique for the early detection of coronary atherosclerosis, and recent studies have established its prognostic value in asymptomatic individuals. The relationship of coronary artery calcium scores (CAC) to obstructive coronary artery disease (CAD) has been poorly studied but is clinically relevant because it determines which individuals are likely to benefit from revascularization procedures. Hence, we prospectively evaluated the prevalence of myocardial ischemia in asymptomatic patients with cardiovascular risk factors and subclinical atherosclerosis. METHODS AND RESULTS: We studied 864 asymptomatic patients with no previous CAD but with cardiovascular risk factors, referred for electron beam tomography coronary calcium imaging to our institution over an 18-month period. From this group, 220 consecutive patients (85% men; mean age, 61 +/- 9 years; age range, 31-84 years) with moderate to severe atherosclerotic disease (coronary calcium score > or =100 Agatston units) were prospectively evaluated by technetium 99m sestamibi single photon emission computed tomography (SPECT). Patients were followed up (mean follow-up, 14 months) and data regarding their subsequent clinical management recorded. Of the 220 patients, 119 had moderate atherosclerosis (CAC score of 100-400 Agatston units) and 101 had severe atherosclerosis (CAC score > or =400 Agatston units). Abnormal SPECT findings were seen in 18% of patients with moderate atherosclerosis (n = 21) and 45% of patients with severe atherosclerosis (n = 45). Increasing severity of atherosclerosis was related to increasing ischemic burden (summed difference score = 1 +/- 0.2 for CAC score of 100-400 Agatston units and 3.2 +/- 0.5 for CAC score > or =400 Agatston units). In a multivariate linear regression model incorporating risk factors, CAC was the only predictor of silent ischemia. CONCLUSION: In comparison to previously published data, we detected a higher prevalence of silent ischemia even in patients with moderate coronary atherosclerosis (18%). This may reflect the differing risk factor profile of our patient population. When coronary calcium screening is used to preselect asymptomatic patients with cardiovascular risk factors for myocardial perfusion imaging, the optimum coronary calcium score threshold will depend on the population prevalence of risk factors and asymptomatic obstructive CAD.     

微血管密度与胃癌侵袭和转移的关系

目的 :探讨胃癌组织中微血管密度 (MVD)与胃癌生物学行为的关系。方法 :应用免疫组织化学S -P法和抗人因子Ⅷ相关抗原 (FⅧ -RAg)的抗体 ,标记 6 8例胃癌组织中的微血管 ,检测MVD。分析其与胃癌组织学类型、分化水平、浸润深度及淋巴结转移的关系。结果 :6 8例胃癌组织中FⅧ -RAg阳性表达 5 2例 ,占 76 .4 7%。FⅧ -RAg和MVD与分化水平、浸润深度 (P <0 .0 1)及淋巴结转移 (P <0 .0 0 1)密切相关。不同组织学类型中FⅧ-RAg的表达具有显著相关性 (P <0 .0 1) ,而MVD则与胃癌组织学类型无显著差异 (P >0 .0 5 )。 结论 :血管生成对胃癌的发展具有一定作用 ,FⅧ -RAg的检测和MVD可作为判断胃癌恶性程度和预后的生物学指标。     

新疆出血热病毒与流行性出血热病毒的生物学特性比较

本文通过免疫荧光(IF)和电镜(EM)技术对新疆出血热(XHF)及流行性出血热(EHF)病毒的生物学特性在Vero E_6及LLC-MK_2两株传代细胞上进行了比较。两种病毒感染两株细胞后免疫荧光染色有两种不同的荧光形态特征,两种病毒对细胞的敏感性明显不同,XHF病毒在LLC-MK_2细胞上感染后6～8天可出现明显的细胞病变,而EHF病毒感染此细胞后则不引起病变。电镜观察到两种病毒在LLC-MK_2细胞中都能形成与布尼病毒相似的形态特征。     

冷冻(断裂)复型电镜技术的应用

本文简单介绍冷冻断裂复型电镜技术在国内、外的概况,並阐明了该项新技术的独特作用和具体的操作步骤;在制作生物样品时所需的仪器设备和试剂的配制等内容。     

Bone mineral density of the proximal tibia following uncemented arthroplasty

Bone mineral density (BMD, g/cm2) was measured using dual-photon absorptiometry (DPA) in selected areas of the proximal tibia following uncemented PCA knee prosthesis. In nine patients with 14 alloplastic operations, measurements were taken at 3-6-month intervals for the first 3 1/2 years after operation. There was a significant increase in BMD of about 15% during the first 6 months after operation. The following year it remained increased, although not significantly, compared with the initial values, then gradually diminished. Increased bone density after arthroplasty may be explained mainly by stimulation of bone formation from weight bearing due to improved walking ability. Stress shielding of the proximal part of the supporting tibial bone did not seem to occur.     

The cholinergic innervation of the rat fascia dentata: identification of target structures on granule cells by combining choline acetyltransferase immunocytochemistry and Golgi impregnation

A monoclonal antibody against choline acetyltransferase (ChAT), the acetylcholine-synthesizing enzyme, was used to study cholinergic synapses on identified (Golgi stained) granule cells in the rat fascia dentata. Choline acetyltransferase immunocytochemistry was applied to 40-microns Vibratome sections cut perpendicular to the longitudinal axis of the hippocampus. Light microscopy revealed fine varicose ChAT-immunoreactive axons in all layers of the fascia dentata, i.e., in the stratum moleculare, the stratum granulosum, and the subgranular polymorph zone. Most fibers were observed in the vicinity of granule cell bodies where they ran mainly parallel to the granular layer. Next, the immunostained Vibratome sections were sandwiched between small pieces of Parafilm and piled to form a block that was covered with agar and Golgi stained. After that, the sections were separated by cutting away the agar and removing the Parafilm. Sections containing well-impregnated granule cells were gold-toned (Fairén et al., '77), embedded in Araldite, and subjected to ultrathin sectioning for electron microscopy. A total of 14 gold-toned granule cells were examined in the electron microscope for synaptic contacts with cholinergic afferents. Choline acetyltransferase-immunoreactive axon terminals were observed that established symmetric synaptic contacts with the cell bodies and dendritic shafts of the gold-toned identified granule cells. Two types of contact were observed on spines arising from gold-toned granule cell dendrites. Immunoreactive terminals established asymmetric synaptic contacts with the head of small spines and symmetric contacts with the stalk of large, complex spines. The boutons forming asymmetric synaptic contacts with the cup-shaped spine head of the complex spines were not found to be immunoreactive. Our results demonstrate that cholinergic fibers to the rat fascia dentata establish characteristic types of synaptic contact with different postsynaptic elements of granule cells, suggesting a complex function of this afferent system.