激光共聚焦显微成像技术对亚细胞位点光动力损伤的动态观察

目的探讨应用激光共聚焦显微成像技术对光动力效应所致亚细胞位点损伤进行动态观察的可行性。方法实验分为HMME＋Rhodamine-123组、单加HMME组、单加Rhodamine-123组和单纯细胞组，后3组作为对照组。传代培养鼠肺毛细血管内皮细胞，将血卟啉单甲醚（HMME）与内皮细胞共同孵育24h后，加入细胞探针Rhodamine-123对线粒体进行染色。应用激光共聚焦显微镜并采用细胞器-细胞荧光强度比值法对HMME进行亚细胞定位，随后采集Rhodamine-123的荧光图像动态序列。结果HMME＋Rhodamine-123组的Rhodamine-123荧光图像在光照过程中逐渐发生变化：典型的线粒体形态特征逐渐消失，并伴有荧光强度下降，荧光在胞浆内趋于弥散分布，细胞核内出现Rhodamine-123的荧光；而单加Rhodamine-123组的Rhodamine-123荧光图像未发生明显变化。结论应用激光共聚焦显微成像技术能实现亚细胞水平光敏损伤位点的动态光学检测，线粒体和核膜可能是HMME介导的光动力损伤亚细胞位点。     

应用超高灵敏度荧光显微成像及共聚焦显微成像观察光敏剂细胞内分布的对比研究

背景：光敏剂亚细胞分布研究多采用激光共聚焦显微镜来开展。激光共聚焦显微成像系统能够获得细节清晰的细胞断层荧光图像，可以进行精确的光敏剂亚细胞定位。但其样品准备过程复杂，费用不菲，而且对于荧光效率较低的光敏剂探测难度很大。目的：应用带像增强器的冷电感耦合装置(intensified charge-coupled device，ICCD)的超高灵敏度荧光显微成像系统及共聚焦显微成像进行光敏剂细胞内分布的对比研究，深入探讨光动力疗法的损伤机制，为拓展其在临床康复治疗领域的适应证奠定实验基础。设计：非随机非对照的实验研究。地点、材料和干预：实验地点为北京理工大学光电工程系实验室。传代培养内皮细胞、食管癌细胞和肺癌细胞，将不同浓度血卟啉单甲醚(hematoporphrin monomethyl ether，HMME)与细胞共同孵育不同时间。采用荧光显微镜及ICCD组成的荧光显微成像系统采集不同浓度及不同孵育时间条件下HMME的荧光图像，并采用计算机图像处理技术进行图像增强、滤波后计算其细胞浆与细胞核的平均荧光强度比值。同时应用激光共聚焦显微镜图像采集进行对比。主要观察指标：不同荧光显微成像系统采集到的细胞的光敏剂荧光图像的细节特点及胞浆与胞核区域光强比值的对比观察。结果：HMME浓度为5mg／L时，荧光显微镜采集到HMME的荧光图像；HMME浓度升高到160mg／L，激光共聚焦显微镜获得HMME的荧光图像。两组图像的特点都为胞浆中荧光强度较高，细胞核区荧光较弱；细胞浆与细胞核的比值约为2～3：1。结论：荧光显微镜和ICCD采集细胞内光敏剂的荧光图像灵敏度高，方法可靠、实用，可用来对荧光效率较低的光敏剂进行细胞内分布研究。HMME较多分布在细胞质中，细胞核吸收较少。从而得出HMME介导的内皮细胞及肺癌细胞光动力损伤的初次损伤位点可能是细胞浆内多种细胞器。     

两种荧光显微成像系统亚细胞定位的对比

目的:应用高分辨率荧光显微成像系统采集细胞器探针图像,并与激光共聚焦显微成像系统进行对比。方法:实验于2003-05/2004-01在解放军总医院完成。①实验材料:鼠肺毛细血管内皮细胞株(1H11)由上海复旦张江生物公司提供;荧光探针Rhodamine-123,Lucifer Yellow,DiOC6[3],BODIPY(美国Sigma公司)。②细胞培养及荧光探针染色:细胞培养采用含体积分数为0.2胎牛血清的低糖DMEM培养基,密度5×107L-1。选择Rhodamine-123作为细胞线粒体特异性荧光探针,选择DiOC6[3]作为细胞内质网特异性荧光探针,选择BODIPY作为细胞高尔基体特异性荧光探针,选择Lucifer Yellow作为细胞溶酶体探针。前3个探针在完全避光条件下与培养的细胞共同孵育0.5h,后者则共同孵育15h。③高分辨率荧光成像系统的图像采集:线粒体荧光图像采集,选取经Rhodamine-123共孵育完成的细胞,选择激发滤色镜为BP460-490,吸收滤色镜为BA515,分光镜为DM500,另加一绿通道液晶滤光片,激发出Rhodamine-123的荧光。电荷耦合器件采集图像并送入计算机。重复上述步骤,采用DiOC6[3]标记内质网,BODIPY标记高尔基体,Lucifer Yellow标记细胞溶酶体,激发条件同Rhodamine-123。分别采集同一视野靶细胞DiOC6[3]、BODIPY或Lucifer Yellow的荧光图像,完成全部图像采集并储存在计算机中。④激光共聚焦显微成像系统的图像采集:选择经4种探针染色的靶细胞,使用氩离子激光器在488nm激发Rhodamine-123,Rhodamine-123荧光通过配置有530/60-G发射滤光片的通道1探测。重复上述步骤,在488nm激发DiOC6[3]和BODIPY,在457nm激发Lucifer yellow,3种荧光均由通道1探测,后2个探针的发射滤光片的配置为515/30-G,DiOC6[3]选择530/60-G。由光电倍增管接收信号并传输入计算机成像。结果:①高分辨率荧光成像系统所采集图像,靶细胞中由荧光探针Rhodamine-123染色的线粒体呈多个典型的小棒状或卵圆状,聚集在核周;Lucifer yellow染色的溶酶体呈多个非对称球型,在胞浆内随机分布,颗粒尺寸通常大于线粒体;荧光探针DiOC6[3]着色的内质网占据胞浆的很大空间,以囊状聚集为特征;BODIPY特异性地结合在高尔基体上,荧光图像显示围绕在细胞核周围呈条索状。②与高分辨率荧光成像系统比较,激光共聚焦显微成像系统所采集的图像其荧光强度基本相同,但分辨率低、细节显示模糊、胞浆中细胞器的准确分布信息和形态特征显示效果欠佳。结论:两种荧光显微成像系统均可采集到细胞器探针的荧光图像。但高分辨率荧光成像系统采集的荧光图像具有细节清晰、分辨度高、准确显示胞浆中细胞器的分布信息和形态特征等优点。     

应用超高灵敏度荧光显微成像及共聚焦显微成像观察光敏剂细胞内分布的对比研究

背景光敏剂亚细胞分布研究多采用激光共聚焦显微镜来开展.激光共聚焦显微成像系统能够获得细节清晰的细胞断层荧光图像,可以进行精确的光敏剂亚细胞定位.但其样品准备过程复杂,费用不菲,而且对于荧光效率较低的光敏剂探测难度很大.目的应用带像增强器的冷电感耦合装置(intensified charge-coupled device,ICCD)的超高灵敏度荧光显微成像系统及共聚焦显微成像进行光敏剂细胞内分布的对比研究,深入探讨光动力疗法的损伤机制,为拓展其在临床康复治疗领域的适应证奠定实验基础.设计非随机非对照的实验研究.地点、材料和干预实验地点为北京理工大学光电工程系实验室.传代培养内皮细胞、食管癌细胞和肺癌细胞,将不同浓度血卟啉单甲醚(hematoporphrin monomethyl ether,HMME)与细胞共同孵育不同时间.采用荧光显微镜及ICCD组成的荧光显微成像系统采集不同浓度及不同孵育时间条件下HMME的荧光图像,并采用计算机图像处理技术进行图像增强、滤波后计算其细胞浆与细胞核的平均荧光强度比值.同时应用激光共聚焦显微镜图像采集进行对比.主要观察指标不同荧光显微成像系统采集到的细胞的光敏剂荧光图像的细节特点及胞浆与胞核区域光强比值的对比观察.结果HMME浓度为5 mg/L时,荧光显微镜采集到HMME的荧光图像;HMME浓度升高到160 mg/L,激光共聚焦显微镜获得HMME的荧光图像.两组图像的特点都为胞浆中荧光强度较高,细胞核区荧光较弱;细胞浆与细胞核的比值约为2～31.结论荧光显微镜和ICCD采集细胞内光敏剂的荧光图像灵敏度高,方法可靠、实用,可用来对荧光效率较低的光敏剂进行细胞内分布研究.HMME较多分布在细胞质中,细胞核吸收较少.从而得出HMME介导的内皮细胞及肺癌细胞光动力损伤的初次损伤位点可能是细胞浆内多种细胞器.     

光动力治疗促进反义bcr-ab1寡核苷酸K562细胞核内转染的实验观察

目的：体外研究光动力治疗(photodynamic therapy，PDT)对b3a2型反义bcr．abl寡核苷酸(assntlsense oligonucleotides，ASO)慢性髓细胞性白血病(chronic myelogenous leukemia，CML)细胞株K562转染的影响，为PDT。合并反义技术应用于CML患者治疗以及体外骨髓净化提供实验基础。方法：半导体激光，波长650nm，剂量9J／cm^2垂直照射。光敏剂为血卟啉单甲醚(hematoperphyrin monomethyl ether,HMME)，采用体外细胞培养技术，荧光显微镜检测荧光标记的反义bcr-abl寡核苷酸在K562细胞中的分布情况。结果：PDT 6h后，细胞内荧光物质主要以明亮的点状聚集在胞核和部分胞浆中，而直接转染时，胞内荧光物质则弥散分布于胞浆中。结论：PDT可以使反义ber-ab1寡核苷酸K562细胞转染主要集中于细胞核内。     

问：激光扫描共聚焦显微镜有何优越性？

答：激光扫描共聚焦显微镜（Laser scanning Confocal Microscopy，LSCM）为近代生物医学图像仪器的最重要发展之一。它是在荧光显微镜成像基础上加装激光扫描装置，使用紫外光或可见光激发荧光探针，利用计算机进行图像处理，从而得到细胞或组织内部微细结构的荧光图像。此外。其还可从亚细胞水平观察诸如Ca^2＋、pH值、膜电位等生理信号及细胞形态的变化。     

非小细胞肺癌EML4／ALK融合基因检测

目的建立荧光原位杂交方法（fluorescent in situ hybridization,FISH）检测EML4／ALK融合基因的方法。方法依据ALK和EML4断裂重排方式,设计、制备双色荧光探针。以人外周血培养细胞为检测对象评价EML4／ALK融合基因检测探针的灵敏度和特异性,以非小细胞肺癌（non-small cell lungcancer,NSCLC）组织样本为对象进行EML4／ALK融合基因检测探针的性能评价。结果本研究建立的FISH检测方法在人外周血培养细胞检测中的特异性和灵敏度均达到100％。在对10例NSCLC腺癌患者石蜡包埋组织样本检测中,筛查出1例EML4／ALKFISH阳性样本。结论本研究制备的双色荧光探针可用于非小细胞肺癌EML4／ALK融合基因检测。     

大肠早癌自体荧光检测系统研究Ⅰ．大肠癌自体荧光光谱诊断研究

本课题采用氙灯作为激发光源，用固定波长同步荧光扫描法对离体大肠癌及正常大肠组织固体样品进行自体荧光扫描分析，样品来自于３７例大肠癌手术后标本。结果显示：大肠癌组织与正常大肠组织其荧光光谱存在差异，表现在４７０±１．１ｎｍ及６４４．３±５．７ｎｍ处荧光波峰强度差异，前者癌组织荧光强度低于正常大肠组织，而后者癌组织荧光强度高于正常大肠组织（Ｐ＜０．０５），另外４７０±１．１ｎｍ处为荧光波形变异，正常大肠组织表现为２种波形变化（即波型Ⅰ、波型Ⅱ、而大肠癌却只表现为波型Ⅰ）。用二值回归法在ＳＡＳ软件包中建模，确立判别函数，并对新样品进行判别分析，结果：敏感性、特异性、阳性预期值分别为９５．７％、７９．３％、７８．６％。根据上述结果，我们认为大肠癌组织其荧光光谱分析能从正常大肠组织中区分。     

细胞涂片间期荧光原位杂交在血液系统疾病中的应用

本研究探讨细胞涂片间期荧光原位杂交(FISH)在血液系统疾病中的临床应用价值。以BCR/ABL双色双融合探针检测Ph染色体为例,同时应用了甲醇/冰醋酸处理的外周血、骨髓细胞涂片法间期FISH与常规细胞悬液滴片法间期FISH,检测20例经证实存在Ph染色体的慢性髓系白血病或急性淋巴细胞白血病患者。结果表明:与常规间期FISH相比,细胞涂片法间期FISH能够获得同样可靠的检测结果。结论:细胞涂片法间期FISH的检测结果可靠,检测周期短,易于进行回顾性研究,值得进一步开展。     

光动力治疗促进反义bcr-abl寡核苷酸K562细胞核内转染的实验观察

目的:体外研究光动力治疗(photodynamictherapy,PDT)对b3a2型反义bcr-abl寡核苷酸(assntisenseoligonucleotides,ASO)慢性髓细胞性白血病(chronicmyelogenousleukemia,CML)细胞株K562转染的影响,为PDT合并反义技术应用于CML患者治疗以及体外骨髓净化提供实验基础。方法:半导体激光,波长650nm,剂量9J/cm2垂直照射。光敏剂为血卟啉单甲醚(hematoperphyrinmonomethylether,HMME),采用体外细胞培养技术,荧光显微镜检测荧光标记的反义bcr-abl寡核苷酸在K562细胞中的分布情况。结果:PDT6h后,细胞内荧光物质主要以明亮的点状聚集在胞核和部分胞浆中,而直接转染时,胞内荧光物质则弥散分布于胞浆中。结论:PDT可以使反义bcr-abl寡核苷酸K562细胞转染主要集中于细胞核内。     

Hematoporphyrin Monomethyl Ether Enhances the Killing Action of Ultrasound on Osteosarcoma In Vivo

Objectives. Osteosarcoma is one of the most common malignant cancers afflicting young adults. Ultrasound is a new therapeutic modality for controlling malignant cancers. Enhancing the efficacy of ultrasound treatment will improve clinical outcomes. The aim of this study was to investigate the killing action of ultrasound on osteosarcoma enhanced by hematoporphyrin monomethyl ether (HMME) in vivo. Methods. An animal model of an osteosarcoma xenograft was set up to investigate the inhibitory effect of sonoactivating HMME on osteosarcoma. High-performance liquid chromatography was used to analyze the time course of HMME in the osteosarcoma xenograft. Three hours after intravenous (IV) administration of HMME, ultrasound radiation was administered in the osteosarcoma xenograft. On day 7 after ultrasound radiation, the tumor volume and weight were measured and calculated for effect assessment, hematoxylin-eosin staining for histopathologic examination, immunohistochemical staining for proliferating cell nuclear antigen (PCNA) expression, and terminal deoxyuridine nick end-labeling (TUNEL) staining for apoptosis examinations. Results. The peak value of HMME in osteosarcoma tissues increased with time after IV administration of HMME and reached a plateau at 3 hours. The increasing rates of the tumor volume and weight in the control group were very fast, but the increasing rates in the ultrasound-alone group were slower, and those in the ultrasound-HMME group were the slowest throughout the observation period. There was a significant difference between the HMME-ultrasound, ultrasound-alone, and control groups (P < .01). Hematoxylin-eosin staining showed that some cells had typical cell death such as pyknosis and nuclear fragmentation after ultrasound radiation alone. More cells with pyknosis, nuclear fragmentation, and even karyolysis were found after HMME-ultrasound treatment. Immunohistochemical staining showed that the percentage of PCNA-positive cells decreased and that of TUNEL-positive cells increased after ultrasound treatment alone, and the changes in the PCNA- and TUNEL-positive percentages were significantly enhanced by pretreatment with HMME (20 mg/kg, IV) for 3 hours and ultrasound radiation (10.5 MHz) for 120 seconds at an intensity of 0.8 W/cm² (P < .05). Conclusions. Hematoporphyrin monomethyl ether pretreatment could substantially enhance the growth inhibition of ultrasound on osteosarcoma, which suggests that HMME is an efficient sonosensitizer, and ultrasound radiation with HMME could be developed as a new modality for treating osteosarcoma.     

SonoLiver^R CAP造影新软件开拓乳腺肿块诊断的新视野

目的通过新的 SonoLiver^R CAP伪彩色造影分析软件对乳腺肿块的应用研究,探讨乳腺良恶性肿块的诊断新方法。方法对本组2006年11月至2009年5月经手术病理证实64个乳腺肿块常规检查后进行超声造影检查,并以 SonoLiver^R CAP伪彩色造影分析。结果64例乳腺肿块CAP软件分析显示红色为主的高增强区37例,蓝色为主的低增强区27例,肿瘤性质与回声增强的关系,恶性肿瘤中76．5％为高增强,23．5％为低增强,良性肿瘤中26．9％为高增强,73．1％为低增强。乳腺炎症均显示为高增强。高增强肿块以恶性为主,低增强肿块以良性为主,恶性组中以整体不规则团块状及散在分布的斑片状为主,达84．6％,良性组以点线状为主,达85．7％。而且峰值强度时相测定高增强紫红色区域占整个肿块造影区域的面积百分比不同：恶性（58．39±18．20）％,良性（32．78±16．30）％（P〈0．05）。结论 SonoLiver^R CAP造影分析软件能将组织结构造影的灰阶高低用彩色编码,清晰显示,直观,简单,开拓了乳腺肿块诊断的新视野。     

病房护理活动调查分析

目的通过分析某综合医院病房护理工作内容厦分类构成厦病房不同岗位的人均任务负荷，为人员配备与绩效考核提供参考依据。方法调查对象为某医院病房所有护理人员210人。调查病房护理人员所从事的各项护理活动内容、耗时与频次。主要采用工作日志法，结合专家咨询法。结果病房护理任务量中白班是夜班的1倍多。病房白班护理任务中直接护理、间接护理任务耗时都略低于夜班，而白班的非护理任务耗时高于夜班。内、外科护理任务各大项目的构成排序基本一致，而ICU和妇产科略有不同，内外科护理任务构成排在首位的是治疗给药，IcU和妇产科则是病情评估。ICU直接护理任务耗时比例高于其它病房，非护理任务耗时最低。病房各岗的人均每班护理任务负荷，夜班与责护人均每班护理任务负荷较高，主班岗则较低。结论该院病房护理任务分类基本合理，但具体任务如治疗给药等应加强调研与改进，以提高效率，减少耗时；部分岗位任务负荷重，需增加护理人员配置。     

Transport of bacterial lipopolysaccharide to the golgi apparatus.

Addition of lipopolysaccharide (LPS) to cells in the form of LPS-soluble (s)CD14 complexes induces strong cellular responses. During this process, LPS is delivered from sCD14 to the plasma membrane, and the cell-associated LPS is then rapidly transported to an intracellular site. This transport appears to be important for certain cellular responses to LPS, as drugs that block transport also inhibit signaling and cells from LPS-hyporesponsive C3H/HeJ mice fail to exhibit this transport. To identify the intracellular destination of fluorescently labeled LPS after its delivery from sCD14 into cells, we have made simultaneous observations of different organelles using fluorescent vital dyes or probes. Endosomes, lysosomes, the endoplasmic reticulum, and the Golgi apparatus were labeled using Texas red (TR)-dextran, LysoTrackertrade mark Red DND-99, DiOC6(3), and boron dipyrromethane (BODIPY)-ceramide, respectively. After 30 min, LPS did not colocalize with endosomes, lysosomes, or endoplasmic reticulum in polymorphonuclear leukocytes, although some LPS-positive vesicles overlapped with the endosomal marker, fluorescent dextran. On the other hand, LPS did appear to colocalize with two markers of the Golgi apparatus, BODIPY-ceramide and TRITC (tetramethylrhodamine isothiocyanate)-labeled cholera toxin B subunit. We further confirmed the localization of LPS in the Golgi apparatus using an epithelial cell line, HeLa, which responds to LPS-sCD14 complexes in a CD14-dependent fashion: BODIPY-LPS was internalized and colocalized with fluorescently labeled Golgi apparatus probes in live HeLa cells. Morphological disruption of the Golgi apparatus in brefeldin A-treated HeLa cells caused intracellular redistribution of fluorescent LPS. These results are consistent with the Golgi apparatus being the primary delivery site of monomeric LPS.     

Use of a blocking antibody method for the flow cytometric measurement of ZAP-70 in B-CLL

BACKGROUND: In this study we developed a method to measure the amount of ZAP-70 [zeta accessory protein] in B-CLL cells without relying on the ZAP-70 expression of patient B or T cells to normalize fluorescence intensity. METHODS: B-CLL cells were fixed with formaldehyde before surface staining with gating antibodies CD19PC5 and CD5FITC. The cells were permeabilized with saponin, and the ZAP-70 antigen was blocked in one tube with unlabeled antibody to ZAP-70 [clone 1E7.2]. Zap-70-PE was then added to this tube. ZAP-70-PE was added to a second tube without unlabeled antibody to ZAP-70. The mean fluorescence intensity of the ZAP-70 in the tube without unlabeled antibody divided by the mean fluorescence intensity of the ZAP-70 in the tube with unlabeled antibody equals the RATIO of total fluorescence to non-specific ZAP-70 fluorescence in the B-CLL cells. In a second method of analysis, a region is created in the histogram showing ZAP-70 fluorescence intensity in the tube with unlabeled antibody to ZAP-70. This region is set to 0.9% positive cells. This same region is then used to measure the % positive [%POS] ZAP-70 cells in the tube without unlabeled antibody to ZAP-70. The brighter the ZAP-70 fluorescence above the non-specific background, the higher the %POS. RESULTS: Due to the varying amount of non-specific staining between patient B-CLL cells and other cells, the blocking antibody method yielded a more quantitative and reproducible measure of ZAP-70 in B-CLL cells than other methods, which use the ratio of B-CLL fluorescence to normal B or T-cell fluorescence. Using this improved method, ZAP-70 was determined to be negative if the RATIO was less than 2:1 and positive if the RATIO was greater than 2:1. ZAP-70 was determined to be negative if the %POS was less than 5% and positive if the %POS was greater than 5%, a cut-off value lower than previous values published, due to exclusion of non-specific staining. Both cut-offs were based upon patient specimen distribution profiling. CONCLUSIONS: Use of a blocking antibody resulted in a robust, reproducible clinical B-CLL assay that is not influenced by the need to measure the amount of ZAP-70 in other cells. ZAP-70 results segre gate patients into indolent and aggressive groups suggested by published clinical outcomes.     

A novel amphiphilic fluorescent probe BODIPY–O-CMC–cRGD as a biomarker and nanoparticle vector

Fluorescent probes have been demonstrated to be promising candidates as biomarkers and biological carriers. Our study focuses on the development of a novel amphiphilic fluorescent probe with good photostability, high water solubility, excellent specificity and promising loading capability for tumor diagnosis and treatment. At first, BODIPY dye and O-carboxymethyl chitosan were prepared via a chemical reaction. Then, the prepared BODIPY dye and cRGD were bonded to O-carboxymethyl chitosan successively via an acylation reaction. Finally, we obtained the desired amphiphilic fluorescent probe: BODIPY–O-CMC–cRGD, which was based on the fluorescence resonance energy transfer (FRET) principle for selective visualization of tumors in vitro. Through a series of experiments, we found that this fluorescent probe possessed better fluorescence characteristics and tumor targeting properties. Simultaneously, by self-assembly, the amphiphilic probe encapsulated the other flexible structure of BODIPY2 and the rigid structure of porphyrin, which formed distinct nanoparticles with different particle sizes. Hence, we could observe different phagocytosis processes of the two nanoparticles in the tumor cells via the fluorescence of dyes by confocal laser scanning microscopy. Therefore, the results suggest that the fluorescent probe has advantages in tumor detection, and the constructed tumor-specific nanoparticles show high clinical potential to be utilized not only in visual and precise diagnosis but also in excellent drug delivery for tumor treatment. Henceforth, we will prepare new targeted and visualized pharmaceuticals by replacing BODIPY2 and porphyrin with antineoplastic drugs for future tumor treatment.

The amphipathic fluorescence probe, BODIPY–O-CMC–cRGD, can be applied in visualized diagnoses and as drug delivery vehicles of visualized therapies in the future.     

快速波谱成像用于急性缺血性脑卒中

目的评价快速波谱成像(TSI)技术用于急性缺血性脑卒中(AIS)的价值。方法对40例AIS患者行TSI及常规多体素化学位移波谱成像(CSI),采用对应频谱拟合脚本获得N-乙酰天冬氨酸(NAA)、肌酸复合物(Cr)、胆碱复合物(Cho)、乳酸(Lac)峰下面积及NAA、Cho、Lac与Cr的比值;将TSI和CSI拟合后获得的NAA和Lac分布伪彩图分割后与弥散加权成像(DWI)相融合,分析其匹配程度。结果相比周边正常脑组织,梗死边缘区NAA稍降低,梗死核心区NAA降低程度大于梗死边缘区,Lac则呈相反趋势;梗死周边正常脑组织存在少量Lac分布。TSI与CSI数据经拟合的代谢物分布差异无明显统计学意义(P均>0.05)。TSI与CSI分别显示33例及32例NAA伪彩图暗区分布小于DWI高信号区,Lac伪彩图高亮区明显大于DWI高信号区。结论采用TSI可在较短时间内完成采集,并获得与常规CSI基本一致的代谢物分布数据。AIS患者代谢物分布伪彩图可能与DWI高信号区不匹配。     

血卟啉单甲醚对人宫颈癌Hela细胞体外影响的研究

目的:初步探讨血卟啉单甲醚(hematoporphyrin monomethyl ether,HMME)介导的光动力疗法(HMME-PDT)对体外培养的人宫颈癌Hela细胞的杀伤效应及影响宫颈癌细胞光动力疗法(photodynamic therapy,PDT)杀伤效应的主要因素。方法:通过采用不同光敏剂浓度、光照能量密度及细胞孵育时间介导的光动力作用于人宫颈癌Hela细胞,MTT法检测24h后的细胞生长抑制率。结果:光敏剂浓度<2.5μg/mL的不同组细胞生长抑制率有明显差异,光照能量密度≥1.5J/cm2及孵育时间≥4h的细胞之间生长抑制率差异无统计学意义。HMME 2.5μg/mL、光照能量密度1.5J/cm2和孵育时间≥4h可能是PDT对Hela细胞杀伤作用的最佳参数。结论:特定光源下,光敏剂的浓度、光照剂量及孵育时间是影响HMME-PDT效应的主要因素。     

声动力学治疗对MDA-MB-231细胞增殖及侵袭能力的影响

目的 探讨超声辐照血卟啉单甲醚和声诺维对乳腺癌细胞株MDA-MB-231增殖及侵袭能力的影响。方法 将对数生长期的MDA-MB-231细胞制成细胞悬液,加入HMME和SonoVue后用超声进行辐照。分3组：对照组,不接受治疗;辐照组1：输出声强0.3 W/cm²,辐照时间60 s;辐照组2：输出声强0.6 W/cm²,辐照时间30 s。采用MTS法绘制细胞生长曲线,平板克隆方法观察细胞形成克隆的能力,并用Transwell小室法检测细胞侵袭能力。结果 对照组的生长曲线陡直,而辐照组的曲线平缓。接种2周后对照组出现肉眼可见的细胞克隆,而辐照组未出现。结晶紫染色后观察,辐照组仅可见由少许细胞聚集成的细胞团。接种后24 h、48 h观察,对照组、辐照组1、辐照组2穿越小室的细胞数分别为42.00±13.52、3.52±1.26、2.75±0.96（P<0.05）及104.25±12.61、36.25±3.40、28.75±4.99（P<0.05）。对照组穿越小室的细胞数显著高于辐照组（P<0.05）。结论 超声辐照联合HMME及SonoVue能有效抑制MB-231细胞增殖,降低细胞形成克隆的能力,并显著降低细胞的侵袭能力。