纳米血管生成素-2小干扰RNA质粒制备及功能研究

[目的]制备血管生成素-2小干扰RNA(Ang-2siRNA)质粒纳米微粒,并观察其基因转染能力和基因沉默效果.[方法]应用分子克隆的方法构建Ang-2siRNA质粒,将其与聚乳酸、O羧甲基壳聚糖用水/油/水(W/O/W)双乳化溶剂蒸发法制备纳米微球,扫描电镜观察其形态和粒径.然后转染原代人脐带静脉内皮细胞,观察纳米Ang-2siRNA微粒干扰Ang-2基因表达的效果及其细胞保护功能.[结果]扫描电镜观察到Ang2siRNA纳米微球呈球形和椭球形,粒径150～200 nm,大小分布均匀,包封完整,分散性良好.有较强转基因能力,良好的RNA干扰效果和血管内皮细胞保护功能.[结论]成功制备纳米Ang-2siRNA微粒,并证实其载基因能力和RNA干扰效果,对血管内皮细胞损伤因素有良好保护作用,为进一步研究心肌梗死的血管修复机制奠定了坚实的基础.     

聚磷酸钙纤维和左旋聚乳酸软骨支架复合自体骨髓间充质干细胞构建组织工程化人工关节软骨

目的应用组织工程学技术，体外初步构建组织工程化人工关节软骨。方法制备三维多孔软骨支架材料CPP／PLLA，体外诱导兔MSCs向软骨细胞表型分化，免疫组织化学染色检测软骨特异性Ⅱ型胶原表达，将诱导细胞与软骨支架材料CPP／PLLA复合，体外培养构建人工关节软骨，1周后终止培养，扫描电镜观察组织工程化人工软骨的微观结构；同时将构建人工软骨移植于兔大腿皮下，3周后处死动物，甲苯胺蓝染色观察。结果扫描电镜观察可见该复合材料CPP／PLLA为高孔隙率的网状、连通、微孔结构，微孔分布均匀，孔径大小为300～400Ⅳn之间；兔MSCs经体外软骨表型定向诱导后，Ⅱ型胶原免疫组化染色阳性。诱导后的MSCs可在支架材料内良好贴附生长，细胞被分泌的胶原基质包裹；从体内获取的培养物组织切片观察可见大量的软骨细胞生成，甲苯胺蓝染色阳性。结论经软骨起源诱导后的MSCs与CPP／PLLA复合培养可以构建自体软骨移植的替代物，为应用软骨组织工程方法修复关节软骨缺损和功能重建提供一种新材料，具有较大的潜在应用价值。     

与甲壳素缝线体外复合培养大鼠成肌细胞株L6细胞的形态学

目的：探讨甲壳素作为骨骼肌组织工程支架材料的可行性。 方法：实验于2002-06／12在南方医科大学应用解剖学研究所完成。大鼠成肌细胞株L6与甲壳素医用缝合线体外复合培养。倒置相差显微镜下观察细胞的形态和排列情况。并分别在体外复合培养的第1，2，3，4，5，6天，取出细胞支架复合体，扫描电镜观察成肌细胞的形态和排列情况。 结果：①扫描电子显微镜观察显示，甲壳素医用可吸收缝线的表面比较粗糙。②倒置相差显微镜下可见大鼠L6细胞与甲壳素制备的医用可吸收缝合线间黏附良好，细胞沿缝线排列呈现串珠样，并可以见到细胞聚集现象。③扫描电子显微镜观察，接种24h后，细胞主要为圆球形，随后细胞迁移、伸展、并见突起；第3天，成肌细胞主要为梭形，并见融合趋势；4d后，细胞沿材料纵轴排列并相互融合，并见肌小管样结构形成及细胞外基质分泌。 结论：甲壳素具有作为支架以构建组织工程化骨骼肌的潜能，支架的平行排列有助于工程化骨骼肌纤维良好方向性的形成。     

乳腺良恶性肿块的高频彩色多普勒超声分析

目的:探讨彩色多普勒多参数分析对乳腺良、恶性肿块鉴别诊断的价值。材料和方法:应用高频彩色多普勒对96例104个乳腺实质肿块进行扫查,观察肿块的彩色多普勒血流信号及多普勒血流频谱形态,与手术病理结果对照分析。结果:64.4%的良性肿块可检出血流信号,血流信号多为Ⅰ~Ⅱ级,良性肿块中血流频谱以低阻力搏动型及静脉型多见,91.1%的乳腺恶性肿块可检出血流信号,血流信号多为Ⅱ~Ⅲ级,恶性肿块的血流频谱形态较多样化,即同时有多种血流频谱,且以高阻力搏动型和湍流型为多见。结论:高频彩色多普勒超声多参数综合分析有助于临床对乳腺肿块的诊断及鉴别诊断。     

Individualized Mycophenolate Mofetil Dosing Based on Drug Exposure Significantly Improves Patient Outcomes After Renal Transplantation

Efficacy and safety of mycophenolate mofetil (MMF) may be optimized with individualized doses based on therapeutic monitoring of its active metabolite, mycophenolic acid (MPA). In this 12-month study, 137 renal allograft recipients from 11 French centers receiving basiliximab, cyclosporine A, MMF and corticosteroids were randomized to receive either concentration-controlled doses or fixed-dose MMF. A novel Bayesian estimator of MPA AUC based on three-point sampling was used to individualize doses on posttransplant days 7 and 14 and months 1, 3 and 6. The primary endpoint was treatment failure (death, graft loss, acute rejection and MMF discontinuation). Data from 65 patients/group were analyzed. At month 12, the concentration-controlled group had fewer treatment failures (p = 0.03) and acute rejection episodes (p = 0.01) with no differences in adverse event frequency. The MMF dose was higher in the concentration-controlled group at day 14 (p < 0.0001), month 1 (p < 0.0001) and month 3 (p < 0.01), as were median AUCs on day 14 (33.7 vs. 27.1 mg*h/L; p = 0.0001) and at month 1 (45.0 vs. 30.9 mg*h/L; p < 0.0001). Therapeutic MPA monitoring using a limited sampling strategy can reduce the risk of treatment failure and acute rejection in renal allograft recipients 12 months posttransplant with no increase in adverse events.     

Adjuvant chemotherapy in completely resected non-small-cell lung cancer.

Surgery alone has long been the standard treatment for patients with operable non-small-cell lung cancer (NSCLC). However, despite complete resection, 5-year survival rates have been disappointing, with about 50% of patients eventually suffering relapse and death from disease. Randomized trials conducted in the 1980s hinted at a survival benefit for postoperative cisplatin-based regimens, but they were underpowered. A meta-analysis published in 1995 found a nonsignificant 13% reduction in the risk of death associated with cisplatin-based chemotherapy, with an increase of survival of 5% at 5 years. This led to renewed interest in adjuvant chemotherapy in resected NSCLC. Thousands of patients have been included in a new generation of randomized trials in the last 10 years. Most of these recent studies have now been reported and several have demonstrated a clear survival advantage for patients treated with platin-based adjuvant therapy. These results also suggest a greater benefit with modern two-drug regimens. In view of the most recent data, postoperative platin-based chemotherapy can now be considered the standard of care for completely resected NSCLC patients with good performance status.     

Measurement of the residual radioactivity induced in the front foil of a target assembly in a modern medical cyclotron.

The levels of residual radioactivity induced in Havar foils at the entrance of a high-pressure 18O-enriched water target used for the production of 18F- in a medical cyclotron with 16 MeV protons have been determined using high-resolution gamma-ray spectrometry. Whole body and skin dose rates arising from exposure to these foils during their periodic replacement have been estimated. The results indicate that irradiated foils do not represent a significant radiological hazard for the cyclotron operating staff and that no waste disposal difficulties should be encountered after an appropriate 'cooling' period of 2 years.     

Does systemic inflammation trigger local exercise-induced oxidative stress in COPD?

Inflammatory abnormalities may be involved in the inadequate basal oxidant/antioxidant balance and local exercise-induced oxidative stress in chronic obstructive pulmonary disease (COPD) patients. The time course of oxidative stress and inflammation was investigated in 10 COPD patients and seven healthy subjects before and after local dynamic quadriceps endurance exercise at 40% of maximal strength. Venous samples were collected before, immediately after and up to 48 h after exercise. At rest, levels of an oxidant released by stimulated phagocytes, the superoxide anion, were significantly higher in patients, as were plasma levels of C-reactive protein, tumour necrosis factor-alpha and interleukin-6, inflammatory markers. An inverse relationship was found between baseline C-reactive protein levels and endurance time in patients. Six hours after exercise, superoxide anion release and levels of protein oxidation products, an index of oxidative stress, increased similarly in both groups, whereas thiobarbituric acid reactive substance levels, another index of oxidative stress, increased significantly only in patients. Plasma nonenzymatic antioxidant and inflammatory cytokine levels were unchanged by the exercise protocol. The increased baseline systemic inflammation in chronic obstructive pulmonary disease patients could be related to disturbed oxidant/antioxidant balance, and, together, these may have triggered the exercise-induced oxidative stress. The absence, however, of local exercise-induced systemic inflammation suggests that additional mechanisms explain local exercise-induced oxidative stress.     

Diagnosis of diffuse and localized arrhythmogenic right ventricular dysplasia by gated blood-pool SPECT.

This study aimed to assess the ability of global and local systolic parameters measured with gated blood-pool SPECT (GBPS) to diagnose and characterize the severity of diffuse or localized arrhythmogenic right ventricular dysplasia (ARVD). METHODS: Fifty-nine subjects with symptomatic ventricular arrhythmias were prospectively included in the study. With the International Society and Federation of Cardiology criteria for ARVD as a gold standard, these subjects were classified as subjects without ARVD (21 control subjects) and patients with localized ARVD (16 patients) or diffuse ARVD (22 patients). Right ventricular volumes, right ventricular ejection fractions (EF), the SD of local EF (sigma-EF), and the SD of the local times of end systole (sigma-TES) were computed from GBPS data and compared among the groups in the study population. RESULTS: sigma-EF did not differ between control subjects and patients with diffuse or localized ARVD. Right ventricular EF and volumes differed between patients with diffuse ARVD and control subjects, with similar areas under the receiver-operating-characteristic curves, but right ventricular EF and volumes failed to differentiate patients with localized ARVD. In contrast, sigma-TES differed between patients with diffuse or localized ARVD and control subjects. Regression analysis showed that the systolic parameter most strongly associated with the diagnosis of ARVD was sigma-TES. The probabilities of a randomly chosen patient in the diffuse ARVD group and of a randomly chosen patient in the localized ARVD group having sigma-TES values greater than that of a randomly chosen control subject were 98.5% and 96.7%, respectively. For the diagnosis of localized ARVD, a threshold of 80 ms for sigma-TES corresponded to sensitivity, specificity, and positive and negative predictive values of 100%, 81%, 80%, and 100%, respectively. CONCLUSION: With GBPS, both diffuse ARVD and localized ARVD can be accurately diagnosed by computing sigma-TES for all of the pixels on the surface of the right ventricle.