抗菌处理对猪肺动脉瓣膜细胞活性及组织结构的影响

目的 探讨抗菌处理对肺动脉瓣膜细胞活性和组织结构的影响因素及优化同种瓣膜的制备方法。方法 3组猪肺动脉瓣叶(n=6)浸泡在含抗生素的DMEM液中，在不同温度下孵育6h或24h。然后通过XTT比色法测定瓣膜细胞活性，用免疫组织化学荧光染色检测瓣膜细胞和细胞外基质，并行光镜观察。结果 抗菌时间为24h，37℃抗菌组与4℃抗菌组的瓣膜细胞活性与瓣膜组织结构无明显区别。抗菌6h组的瓣膜细胞活性与瓣膜组织结构明显优于抗菌24h组。结论 抗菌时间为24h时，抗菌温度对瓣膜细胞活性及结构完整性无明显影响。抗菌时间缩短至6h，有利于瓣膜细胞活性与结构完整性的保持。     

获得性纯红再障患者外周血T细胞亚群分析

目的　本研究通过分析获得性纯红再障患者外周血T细胞亚群 ,进一步探讨该病的细胞免疫发病机制。方法　用抗人CD3、CD4、CD8、IFN γ及IL 4单抗结合人外周血单个核细胞 ,流式细胞术分析单个核细胞中CD3 +、CD4+、CD8+细胞数Th1、Th2、Tc1、Tc2细胞数以及CD4+/CD8+、Th1/Th2、Tc1/Tc2比值。结果　贫血组患者 ,外周血CD3 +细胞和CD8+细胞明显增加 (P <0 .0 1) ,CD4+/CD8+比例倒置 (P <0 .0 5 ) ;Tc2细胞明显增加 (P <0 .0 5 ) ,Tc1/Tc2比值降低(P <0 .0 5 ) ,治疗后血象恢复的患者各增高的亚群细胞均有所恢复 ,接近正常。结论　在获得性纯红再障患者T细胞明显增高 ,其中主要是CD8+细胞 ,而在CD8+细胞中 ,是Tc2细胞发挥了重要作用。经治疗有效的患者 ,其增高的T细胞亚群降低 ,提示以纠正病人细胞免疫异常为主的治疗方案有益于获得性纯红再障的治疗。     

登革病毒感染后小鼠细胞免疫功能的动态变化

从登革Ⅱ型病毒感染成年ＢＡＬＢ／ｃ小鼠为模型，观察感染后不同时间机体细胞免疫反应的变化。结果表明：感染后７－１４ｄ免疫功能处于激活状态，表现为腹腔巨噬细胞的吞噬功能和脾淋巴细胞对ＣｏｎＡ的反应及ＩＬ－２产生水平都明显高于对照组，感染２１ｄ后免疫功能则转为抑制状态，上棕免疫反应明显低于对照组，脾中的Ｌ３Ｔ４＾＋细胞亚群的百分比逐渐下降；相反，ＬＹＴ２＾＋细胞亚群的百分比逐渐升高，这种抑制现象可维持到     

Induction of Donor-Specific Allograft Tolerance by Short-Term Treatment with LF15-0195 After Transplantation. Evidence for a Direct Effect on T-Cell Differentiation

A 20-day treatment with LF15-0195, a deoxyspergualine analog, induced long-term heart allograft survival in the rat without signs of chronic rejection. LF15-0195-treated recipients did not develop an anti-donor alloantibody response. Analysis of graft-infiltrating cells, IL10, TNFalpha, IFNgamma mRNA and iNOS protein expression in allografts, 5 days after transplantation, showed that they were markedly decreased in allografts from LF15-0195-treated recipients compared with allografts from untreated recipients. Surprisingly, spleen T cells from LF15-0195 recipients, 5days after grafting, were able to proliferate strongly in vitro, when stimulated with donor cells, but had reduced mRNA expression for IFNy compared with spleen T cells from untreated graft recipients. Furthermore, when T cells from naive animals were stimulated in vitro, using anti-CD3 and anti-CD28, LF15-0195 also increased T-cell proliferation in a dose-dependent fashion: however, these cells expressed less of the Th1 -related cytokines, IFNgamma and IL2, compared with untreated cells, suggesting that LF15-0195 could act on T-cell differentiation. In conclusion, we show here that a short-term treatment with LF15-0195 induced long-term allograft tolerance, decreasing the in situ anti-donor response, and we illustrate evidence for the development of regulatory mechanisms.     

In vivo MRI of cancer cell fate at the single-cell level in a mouse model of breast cancer metastasis to the brain.

Metastasis (the spread of cancer from a primary tumor to secondary organs) is responsible for most cancer deaths. The ability to follow the fate of a population of tumor cells over time in an experimental animal would provide a powerful new way to monitor the metastatic process. Here we describe a magnetic resonance imaging (MRI) technique that permits the tracking of breast cancer cells in a mouse model of brain metastasis at the single-cell level. Cancer cells that were injected into the left ventricle of the mouse heart and then delivered to the brain were detectable on MR images. This allowed the visualization of the initial delivery and distribution of cells, as well as the growth of tumors from a subset of these cells within the whole intact brain volume. The ability to follow the metastatic process from the single-cell stage through metastatic growth, and to quantify and monitor the presence of solitary undivided cells will facilitate progress in understanding the mechanisms of brain metastasis and tumor dormancy, and the development of therapeutics to treat this disease.     

Reperfused myocardial infarctions on T1- and susceptibility-enhanced MRI: Evidence for loss of compartmentalization of contrast media

The purpose of this study was to characterize the contrast caused by a susceptibility MRI contrast agents, on spin echo T₂-weighted imaging of reperfused myocardial infarction. Our interest in this model focused on the expected requirement that such agents be compartmentalized in the tissue to cause signal loss on spin echo images, a condition which may not be present in reperfused infarcted myocardium. Accordingly, nine rats were subjected to 2 h of left coronary artery occlusion followed by 3 ± 0.5 h of reperfusion prior to administration of contrast media. Three sets of MR images were acquired: (a) baseline axial images at the midventricle, both T₁-weighted (TR/TE = 300/20) and T₂-weighted (TR/TE = 1500/60); (b) T₁-weighted images after administering a T₁-enhancing agent, Gd-DTPA-BMA (0.2 mmol/kg), to document that contrast media is delivered to the reperfused infarction; and (c) T₂-weighted images after administering the susceptibility agent, Dy-DTPA-BMA (1.0 mmol/kg). Gadolinium-enhanced T₁ images depicted reperfused infarction as regions with greatly enhanced signal intensity compared with unin-farcted myocardium, indicating that contrast agent was delivered to the infarcted zone. Dysprosium-enhanced T₁ images depicted the injury as a region of persistent signal intensity relative to depletion of signal in normal myocardium, consistent with failure of the contrast agent to cause signal loss. Similar infarction sizes were observed for unenhanced T₂-weighted images (33 ± 5%), gadolinium-enhanced T₁ weighted images (36 ± 5%) and postmortem staining (30 ± 6%); strong correlations (r > 0.9) were noted in comparisons of these data. Dysprosium-enhanced images exhibited a smaller region of differential signal presumed to be infarction (20 ± 5%, P < 0.05) and weak correlations (r < 0.75) with the other measurements. We conclude that the smaller infarction depicted on dysprosium-enhanced images is a subregion of the true infarction in which myocardial necrosis is sufficiently advanced that the agent is homogeneously distributed throughout all tissue compartments, preventing T₂*-dependent phase loss on spin echo images.     

pH control in rat skeletal muscle during exercise,recovery from exercise,and acute respiratory acidosis

We used ³¹P magnetic resonance spectroscopy to compare the response of rat skeletal muscle to three kinds of proton load. During exercise (tetanic sciatic nerve stimulation), protons from lactic acid were buffered passively and consumed by net hydrolysis of phosphocreatine (PCr). During recovery from exercise, the pH-dependent efflux of protons produced by PCr resynthesis could be partially inhibited by amiloride or 4,4′-diisothiocyanostilbene-2,2′-disulphonate (DIDS), implicating both sodiudproton and bicarbonatelchloride exchange, but was not inhibited by simultaneous respiratory acidosis. In early recovery, up to 30% of proton efflux was mediated by lactatelproton cotransport. During acute respiratory acidosis at rest, the eventual change in muscle pH was consistent with passive buffering and was unaffected by amiloride or DIDS, implying no significant contribution of proton fluxes.     

Anthracycline-induced cardiotoxicity: Overview of studies examining the roles of oxidative stress and free cellular iron

The risk of cardiotoxicity is the most serious drawback to the clinical usefulness of anthracycline antineoplastic antibiotics, which include doxorubicin (adriamycin), daunorubicin or epirubicin. Nevertheless, these compounds remain among the most widely used anticancer drugs. The molecular pathogenesis of anthracycline cardiotoxicity remains highly controversial, although the oxidative stress-based hypothesis involving intramyocardial production of reactive oxygen species (ROS) has gained the widest acceptance. Anthracyclines may promote the formation of ROS through redox cycling of their aglycones as well as their anthracycline-iron complexes. This proposed mechanism has become particularly popular in light of the high cardioprotective efficacy of dexrazoxane (ICRF-187). The mechanism of action of this drug has been attributed to its hydrolytic transformation into the iron-chelating metabolite ADR-925, which may act by displacing iron from anthracycline-iron complexes or by chelating free or loosely bound cellular iron, thus preventing site-specific iron-catalyzed ROS damage. However, during the last decade, calls for the critical reassessment of this “ROS and iron” hypothesis have emerged. Numerous antioxidants, although efficient in cellular or acute animal experiments, have failed to alleviate anthracycline cardiotoxicity in clinically relevant chronic animal models or clinical trials. In addition, studies with chelators that are stronger and more selective for iron than ADR-925 have also yielded negative or, at best, mixed outcomes. Hence, several lines of evidence suggest that mechanisms other than the traditionally emphasized “ROS and iron” hypothesis are involved in anthracycline-induced cardiotoxicity and that these alternative mechanisms may be better bases for designing approaches to achieve efficient and safe cardioprotection.     

Magnetic nanoparticle labeling of mesenchymal stem cells without transfection agent: cellular behavior and capability of detection with clinical 1.5 T magnetic resonance at the single cell level.

The purpose of this work was to evaluate the efficacy of labeling human mesenchymal stem cells (hMSCs) by ionic superparamagnetic iron oxide (SPIO) without a transfection agent and verifying its capability to be detected with clinical 1.5 T magnetic resonance (MR) at the single-cell level. Human hMSCs were incubated for 24 h with an ionic SPIO, Ferucarbotran. The labeling efficiency of hMSCs was determined by iron content measurement spectrophotometrically, and the influence of labeling on cell behavior was ascertained by examination of cell viability using the trypan blue exclusion method, cell proliferation analysis using MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay, mitochondrial membrane potential (MMP) change, differentiation capacity, and reactive oxygen species (ROS) production measured by dichlorofluorescein diacetate (DCFDA) fluorescent probe. Labeled hMSCs were scanned under 1.5 T MRI with three-dimensional (3D) and two-dimensional (2D) T(2)-weighted gradient echo (GRE) pulse sequences. Human hMSC labeling without transfection agent was efficient. The iron content in hMSCs was 23.4 pg Fe/cell. No significant change was found in viability, proliferation, MMP change, ROS production, or differentiation capacity. About 45.2% of the hMSCs could be detected using 1.5 T MRI at the single cell level with 3D GRE and four repetitions.     

子宫内膜异位症患者血清及腹腔液中血小板衍生生长因子、血管细胞粘附分子水平的变化及其意义

目的通过检测子宫内膜异位症(EMs)患者血清及腹腔液中血小板衍生生长因子(PDGF)、血管细胞粘附分子-(VCAM-1)浓度,探讨PDGF和VCAM-1在EMs发病中的作用。方法选取手术后病理证实为EMs的40例患者为EMs组,其中I~II期17例,III~IV23例;非EMs组20例作为对照组。应用酶联免疫吸附法(ELISA)检测血清及腹腔液中PDGF和VCAM-1水平。结果EMs组血清及腹腔液中PDGF、VCAM-1水平均明显高于对照组(P<0.01)。随EMs期别的增加,其血清和腹腔液中PDGF、VCAM-1含量呈上升趋势;EMsIII~IV期水平显著高于I~II期(P<0.05)。EMs患者PDGF与VCAM-1呈正相关性(P<0.05)。结论EMs患者PDGF、VCAM-1表达水平升高,在EMs的发生发展中具有重要作用。