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。     

Development and experimental in vivo validation of mathematical modeling of laser coagulation

Most clinical procedures using the laser are based on thermal laser-tissue interactions. The treatment often consists of inducing damage of given degree and extent by heating the tissue. The aim of this study was to develop a model called HELIOS. The ability of HELIOS to predict thermal coagulation was evaluated by comparison with in vivo experimental results. Conversion of laser light in tissue was studied using the beam-broadening model. Temperature was described by the heat conduction equation solved using the finite difference method. The tissue dena-turation was modeled by the Henriques equation leading to the determination of the damage coefficient ω. For a given set of laser and tissue parameters, HELIOS makes a graphic representation of coagulation necrosis and temperature evolution in tissue. HELIOS was validated by experimental studies in vivo on rat liver using a CW Nd:YAG laser, a CO₂ laser, and an argon laser. For given sets of laser parameters, temperature measurements were performed using an infrared camera. Histological examinations were carried out on samples to quantify the depth of coagulation necrosis. Experimental data obtained in vivo were compared with those calculated using HELIOS and similar sets of parameters. The difference between the predicted temperature evolution on tissue surface and that measured by the infrared camera was < 5°C in all cases. The difference between the predicted coagulation necrosis depth and the corresponding experimental one was < 10%. In conclusion, HELIOS allows good prediction of tissue temperature and coagulation necrosis. © 1994 Wiley-Liss, inc.     

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。     

Skin reactions to irritants assessed by non-invasive bioengineering methods

Pathophysiological components of irritant contact dermatitis caused by 3 chemically-different irritants were investigated. 20 healthy volunteers were patch tested with sodium lauryl sulphate, nonanoic acid and hydrochloric acid on the flexor side of the upper arm. The skin response was evaluated after 24, 48 and 96 h by visual scoring and measured by the following bioengineering methods: transepidermal water loss measurement, electrical conductance for measurement of skin hydration, laser Doppler flowmetry for measurement of cutaneous blood flow and 20 MHz ultrasound A-scan for measurement of skin thickness. In spite of homogeneous inflammatory responses, significant differences in the severity of the injury to the skin barrier function caused by the different irritants were found. Also significant differences between irritants were found in the time course of development of maximum irritant reactions. Bioengineering methods indicating inflammatory responses (measurement of blood flow and skin thickness) were helpful in quantifying the irritant response in general, while bioengineering methods indicating epidermal damage (measurement of TEWL and electrical conductance) were helpful in classifying the individual irritants.     

经尿道非接触式激光前列腺切除技术的应用与改进

１９９３年１１月至１９９６年５月应用改进的经尿道非接触式激光治疗前列腺增生１２０例，并对病人进行术前术后的症状评分，尿动力学，残余尿检查，随访病人１－２４个月，结果显示病人无论是症状评分，还是尿流率，残余尿等各项指标均有明显的改善，此种手术具有简单，安全，副作用小，术后恢复快，治疗效果好等优点。     

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。     

母血中胎儿有核红细胞的荧光原位杂交法在产前诊断中的可行性探讨

探讨荧光原位杂交法（FISH）对母血中胎儿有核红细胞（NRBC）进行无创性产前诊断的可行性。20例孕龄15-20周的孕妇外周血经不连续密度梯度离心、制片、显微镜下识别并共计数NRBC及定位，然后行Y染色体的FISH检测。结果发现10例孕男性胎儿的孕妇外周血细胞涂片中每例均有阳性杂交信号出现；阳性率为60％（24/40）。10例孕女性胎儿的孕妇外周血细胞涂片中1例出现阳性杂交信号；阴性率为95％（38/40），假阳性率仅为5％（2/40）。结果提示FISH法对于用母血中分离到的胎儿细胞进行染色体异常的无创性产前遗传学检查具有重要意义。     

用荧光原位杂交技术快速诊断孕早期胚胎染色体数目异常

目的探讨荧光原位杂交(FISH)技术在快速产前诊断孕早期胚胎染色体数目异常中的价值.方法采用13,18,21,X和Y染色体特异性DNA探针,对46例孕47～65 d高危孕妇的绒毛间期细胞进行FISH检测,同时行常规染色体核型分析平行诊断.结果与染色体核型分析结果一致的染色体数目正常43例,异常3例(47,XY 21;47,XY 18和45,X).3例异常核型胚胎经治疗性流产后再分别对其绒毛行常规染色体核型分析,结果与产前诊断相符.结论FISH技术用于产前诊断孕早期胚胎染色体数目异常具有快速、准确等优点.     

MMP-2、TIMP-2在内毒素诱导性葡萄膜炎中的表达

目的　探讨基质金属蛋白酶 2 (MMP 2 )及其组织型抑制剂 (TIMP 2 )在内毒素诱导性葡萄膜炎 (EIU)模型中的表达及意义。方法　 2 0 0 μg伤寒杆菌内毒素注射于SD大鼠双后足垫 ,建立EIU模型。用免疫组织化学方法检测MMP 2、TIMP 2在内毒素注射后不同时间点 (0、 6、 12、 18、 2 4、 4 8、 72、 96h和 7d)的表达。结果　内毒素注射后 6h开始出现炎症 ,于 2 4h炎症达到高峰 ,以后逐渐减轻 ,7d时基本消退。虹膜、睫状体的上皮细胞和渗出的炎性细胞表达MMP 2和TIMP 2。MMP 2在 6h表达开始增高 ,18～ 2 4h达高峰 ,以后逐渐下降。TIMP 2于 12h出现表达 ,4 8h达高峰 ,以后逐渐下降。TIMP 2和MMP 2平均吸光度值的比值与炎症程度呈负相关。结论　MMP 2是与炎症反应相关的调节因子 ,其过量表达参与葡萄膜炎的发病。减低MMP 2的活性或降低MMP 2 /TIMP 2的比值可能为治疗葡萄膜炎的新思路。     

荧光偏振检测HBV DNA C区BCP双突变

目的：建立简便、灵敏的HBV DNA序列中BCP双突变点的检测方法．方法：采用终点终止法和偏振光检测技术进行点突变的检测．首先对HBV C区基因进行PCR扩增，然后用特异探针与扩增产物中待测核苷酸的下游序列杂交，使探针的3’端可以在DNA聚合酶的催化下，依据其互补链上的待测核苷酸连接上一个标有特定荧光素的ddNTP，然后检测该3’端带有荧光素的探针，根据检测到的荧光素种类和偏振光的强度可以判定待测点是何种核苷酸．结果：该方法可以检测出HBV基因序列中BCP双突变核苷酸类型以及检测1个拷贝的模板，并且可以从BCP野性株DNA序列中检出5％BCP双突变DNA序列．结论：该技术可以检测血清中HBV DNA C区BCP双突变．