Laser induced autofluorescence studies of animal skin used in modeling of human cutaneous tissue spectroscopic measurements

BACKGROUND/PURPOSE: Laser-induced autofluorescence spectroscopy provides excellent possibilities for medical diagnostics of different tissue pathologies including cancer. However, to create the whole picture of pathological changes, investigators collect spectral information from patients in vivo or they study different tumor models to obtain objective information for fluorescent properties of every kind of healthy and diseased tissue. Therefore, it is very important to find the most appropriate, and close to the human skin, animal samples from the fluorescence point of view, which will allow the extrapolation of the animal data to human spectroscopic diagnostics. METHODS: In the present work, we examined the autofluorescence properties of different animal skin tissues, which are considered as the most common skin models. A nitrogen laser was used as an excitation source. Samples of healthy mouse, chicken and pig skin in vivo and/or ex vivo were studied and were compared with results obtained from investigations of healthy human skin in vivo. RESULTS AND CONCLUSION: Specific features of the recorded spectra are discussed and the possible origin of the obtained fluorescence signals is proposed. Quantitative evaluation of data extrapolation for each skin type is also depicted.     

特异性荧光偏振免疫法监测521例肾移植后环孢素A全血浓度3275次

目的通过监测肾移植后病人环孢素A(CsA)全血浓度 ,提出CsA在三联免疫抑制用药方案中的理想治疗窗。方法用特异性荧光偏振免疫法测定CsA全血浓度 ,对521例病人监测3275次 ,按术后时间及临床表现分组比较。结果肾移植后<1 ,、1～3、3～6、6～12个月、1～2和>2年的CsA全血谷浓度的理想治疗窗应分别为250～450、200～400、150～300、100～250、100～200和100～180μg/L。结论CsA全血浓度在上述范围内 ,中毒反应和排异反应明显减少     

单波长荧光分光光度计测定细胞内Ca^2＋浓度的方法探索

在使用单波长荧光分光光度计测定细胞内游离钙浓度时，通过快速（４～６ｓ）手动转换激发波长（ＥＸ），分别测定ＥＸ３４０和３８０ｎｍ时的荧光强度变化，并计算出３４０ｎｍ与３８０ｎｍ时的荧光强度比率（Ｒ），然后也采用双波长荧光分光光度计测定细胞内Ｃａ２＋浓度的计算公式计算细胞内游离钙浓度。结果显示单波长荧光分光光度计按比例法测得的细胞内游离Ｃａ２＋浓度与使用双波长荧光分光光度计测得的结果相似。     

Use of commercially available cell culture inserts for primary culture and electrophysiologic studies of guinea pig gastric mucous epithelial cells

Summary The adherence, growth, and electrophysiologic properties of guinea pig gastric mucous epithelial cells were investigated using porous membrane filters. We also tested three commercially available Ussing-type chambers that were designed to be used with the various porous membrane supports. Overall, the 0.45-µm Falcon-Cyclopore porous membrane was found to be very favorable for the consistent attachment and growth of our cells. This same filter also gave good results in the detection of periodic acid Schiff-positive mucous glycoprotein and Nile red neutral lipid fluorescence in the gastric mucous cells. Our cells grew poorly on collagen-coated Costar-Snapwells and Millipore Millicell-CM porous filters. For measurement of transepithelial potential difference resistance, and short-circuit current, the Costar-Snapwell with the Costar-Snapwell Diffusion-chamber system was superior in design and operation when compared to the Costar Transwell-COL, Falcon-Cyclopore, or Anotec-Anocell porous inserts used with conventional Ussing-chambers. The gastric mucous cells grew best on ICN-Cellagen membranes, but these filters routinely detached from their plastic holder and therefore could not be used for Ussing-chamber studies. The large 24.5-mm, 0.40-µm pore size Costar-Transwell-COL and the 24.1-mm, 0.45-µm Falcon-Cyclopore membranes gave good results when used in a modified horizontal-chamber for microelectrode analysis of membrane potentials and resistances of the gastric mucous cell monolayers.     

荷瘤鼠乳腺癌组织的飞秒双光子荧光诊断

目的：检测荷瘤鼠乳腺癌组织的双光子荧光光谱。方法：乳腺癌荷瘤鼠腹腔注射血卟啉（HpD)。以飞秒激光照射小鼠的癌组织部位及周围正常组织部位。观察HpD双光子荧光光谱。结果：小鼠的癌组织部位可观察到外源性HpD双光子荧光光谱，而正常组织部位未观察到明显的双光子荧光光谱。结论：结论：飞秒激光外源性血卟啉双光子荧光诊断可作为癌症诊断的新方法。     

电镜和荧光显微技术在细胞凋亡研究中的应用

目的：探讨荧光显微技术在检测细胞凋亡中的作用。方法：应用拓扑异构酶Ⅱ抑制剂VP-16及化学毒物叠氮钠分别诱导HL-60细胞凋亡和死亡，用透射电镜和经Hoechst 33258染色的荧光显微镜对凋亡及死亡动态观察和定量分析。结果：HL-60细胞在VP16处理后，荧光显微镜下可见核浓缩、染色质凝集、核碎裂等凋亡特征；而叠氮钠诱导的细胞死亡则出现核溶解、核染色质弥散，但无上述改变；两者电镜的观察与荧光显微镜的改变一致；凋亡细胞及坏死细胞荧光显微镜下呈不同的形态特征。对处理的不同时间点细胞的荧光显微镜观察发现：处理后4h核形态开始变化，有核浓缩的细胞比例在处理8h达高峰，然后下降，核碎裂细胞的比例在24h达高峰，约占80％，表明核浓缩发生在核碎裂之前。结论：荧光显微镜技术可明确观察判断细胞凋亡及坏死，并可进行动态及定量分析，是良好的凋亡检测方法。     

糖尿病肾脏胶原非酶糖化与氨基胍阿斯匹林的治疗作用

本实验系统观察了ＳＴＺ所致糖尿病鼠在１、３、６个月后肾脏组织胶原发生非酶糖化的情况，及氨基胍、阿斯匹林对肾脏胶原非酶糖化的阻断作用。结果表明：各病程糖尿病动物肾组织的胶原含量和非酶糖化早期产物（５－ＨＭＦ）及胶原相联荧光值均明显高于正常对照。氨基胍能够减弱糖尿病动物肾组织胶原含量的增加和荧光产物的生成，但对５－ＨＭＦ含量无影响。阿斯匹林未显示出对胶原非酶糖化的影响。     

外围由小分子修饰的树枝状聚酰胺的合成及其荧光性能研究

合成了1－5代外围由小分子荧光体修饰的树枝状聚酰胺，通过红外，紫外，核磁等表征了其结构，并对其荧光性能进行了研究，修饰后的固体产物的荧光较修饰前粘稠体产物的荧光强得多，树枝状高分子荧光受组分比，代数，溶液浓度，溶剂等因素的影响，有机硅对树状高分子的端氨基与3，5二羟基苯甲酸按1／1摩尔比反应时，其产物荧光最强，对于端氨基与3，5－二羟基苯甲酸摩尔比为1／1的不同代树枝状聚酰胺而言，2代产物荧光最强。