云南白药对血小板活化的研究

为探讨云南白药对血小板活化的影响 ,应用单克隆抗体法 ,并用流式细胞仪对 2 0 6例男性腹股沟斜疝患者血小板活化分子标志物进行检测 ,同时检测血浆凝血物质和 D-二聚体含量。结果显示 :服药组血小板明显活化与对照组比较有显著性差异 ( P<0 .0 5 ) ,凝血物质和 D-二聚体与对照组比较无差异 ( P>0 .0 5 )。结论 :云南白药能增加血小板活化 ,促进局部止血 ,但它不影响凝血物质和 D-二聚体的含量 ,不增加血栓形成的危险。     

脾原发性淋巴瘤超微结构观察与流式细胞分析

目的 探讨诊断电镜（DEM）对脾原发性淋巴瘤的病理分型的超微结构特征。流式细胞术（FCM）测定瘤细胞DNA倍性与恶性程度的关系。方法 应用透射电镜和流式细胞术观察分析41例脾原发性淋巴瘤的超微结构和瘤细胞DNA倍性。结果41例脾原发性淋巴瘤中霍奇金淋巴瘤5例,电镜下R-S细胞呈双叶不规则核和对称致密大核仁,核内常染色质明显,胞质内见中等量线粒体,较长的粗面内质网。非霍奇金B细胞性淋巴瘤21例,裂细胞性5例,细胞核大,核伴有较深的裂沟,胞浆少,胞质见少数肿胀线粒体,无核裂细胞性7例,细胞核规则,呈圆或卵圆型,异染色质多呈条块状,核仁中等大,胞质少,见少量线粒体及长粗面内质网。混合细胞性,瘤细胞大小悬殊,大畸形核较多,核扭曲,不规则深沟及不对称分叶,形成多叶核。非霍奇金T免疫母细胞淋巴瘤11例,瘤细胞较大,核小胞质相对多,胞质透明,核多形性,有的呈圆或卵圆形,核膜薄,有深浅不一的凹陷,可见小核仁。淋巴母细胞性5例,淋巴母曲核性淋巴瘤,细胞为中小形,呈锯齿状核和扭曲状核,异染色质沿核膜聚集,核仁小。毛细胞性淋巴瘤1例,毛细胞的绒毛宽基底,核仁大,胞浆无核糖体板层复合物。组织细胞性淋巴瘤3例,细胞浆丰富,核圆或椭圆形,偶见核膜呈锯齿状或折叠,线粒体和溶酶体丰富,无细胞连接和Birbeck颗粒。FCM     

表皮生长因子影响肿瘤患者舌苔变化的分子机制研究

目的 研究表皮生长因子（EGF）对食管癌细胞Eca-109细胞周期及表皮生长因子受体（EGF－R）表达的影响，探讨EGF促进肿瘤患者舌苔增厚的分子机制。方法 应用流式细胞术，检测EGF－R的表达和细胞周期。结果 EGF能明显促进Eca-109细胞膜上EGF－R的表达，使细胞增殖活性大大增强。结论EGF有是通过促进EGF－R的表达影响舌苔形成。     

Effect of Gardenia extract ZG on the adsorption quantity of herpes simplex virus type 1 （HSV-1）

To observe the effect of Gardenia extract ZG on the adsorption quantity of herpes simplex virus type 1 (HSV-1) so as to explore the mechanism of its antiviral activity, fluorescein isothiocyanate (FITC) was used as the fluorescent probe to label viruses and heparin sodium was used as control. Meanwhile , the effect of Gardenia extract ZG on the adsorption quantity on the surface of Hep-2 cells was determined by flow cytometry. It was demonstrated that adsorption of HSV-1 on the surface of Hep-2 cells exhibited the character of saturation and specificity and heparin sodium could prevent attachment of viruses on these cells. These results are in accord with those reported previously. It was also proved that the manner of drug-use prior to adsorption or simultaneous use of drug and adsorption was better than adsorption prior to drug-use, and the inhibition rates of the former and latter manner were 84. 76% and 82.92% respectively. Three manners of drug-use with Gardenia extract ZG were all effective to reduce the adsorption quantity of viruses, especially the manner of simultaneous use of drug and adsorption with an adsorption inhibition rate of 68.46% . From the above observation, it is apparent that the mechanism of anti-viral activity of Gardenia extract ZG may be via several steps involved in the HSV-1 adsorption.     

应用FISH、FCM检测APL PML/RARA融合基因及临床应用的研究

目的　研究应用荧光原位杂交技术 (FISH)、流式细胞技术 (FCM)检测急性早幼粒细胞白血病 (APL)PML/RARa融合基因及残留白血病细胞的意义。方法　应用CG、M -FISH、FCM对 30例APL患者初发和缓解期的骨髓标本进行分析 ,分别检测t(15 ;17)易位和PML/RARa融合基因及残留白血病细胞的存在。结果　对初发期骨髓标本进行CG和FISH分析发现 ,10 0 %具有t(15 ;17)易位 ,阳性核型占 6 8%～ 10 0 % ,其中有 8例还伴有其他异常 ;10 0 %具有PML/RARa融合基因 ,且阳性中期相的比例高达 96 %～ 10 0 % ;结果具有显著差异 (P <0 0 0 1)。对CR期的骨髓标本进行CG、FISH分析 ,CG为正常核型 ,FISH仍可检出 0 %～ 4 5 %的PML/RARa融合基因。对CR后 12个月的标本进行CG、FISH分析 ,CG均为正常核型 ;FISH仍可检出 0 %～ 37%的PML/RARa融合基因 ,较CR期有所降低。对这 30例APL患者初发期、完全缓解期及完全缓解后 12个月时进行了FCM检测 ,并得出定性结果。完全缓解期检测结果与M -FISH分析一致 ;对完全缓解后 12个月时M -FISH结果进行定性分析发现 30例患者中有 6例为融合基因阴性 ,相应的FCM定性分析结果却发现只有其中的 2例呈现阴性 ,其余 4例仍然为阳性。随访至CR后 2年 ,发现 7例PML/RARa融合基因阳性中期相比例较高 (CG分     

海生素对K562细胞生长及凋亡基因表达的影响

目的 探讨海生素对白血病K562细胞生长及凋亡基因表达的影响. 方法实验分为对照组、10mg/L海生素组和20rag/L海生素组.采用流式细胞术(FcM)、四唑蓝(MTT)和免疫细胞化学法测定海生素对K562细胞周期、细胞生长及细胞凋亡基因bcl-2和bax表达的影响. 结果海生素浓度为10mg/L和20mg/L时,可阻止K562细胞周期于G1/S期;海生素浓度为20mg/L时对K562细胞有抑制作用,在此浓度下随时间的延长细胞存活率有下降趋势;海生素浓度为20mg/L和40mg,L时,可下调凋亡抑制基因bcl-2的表达,上调凋亡促进基因hax的表达.结论 海生素可明显抑制K562细胞的增殖,使其阻滞于细胞周期的G1/S期;海生素还通过减少bcl-2的表达及增加bax的表达从而促进K562细胞的凋亡.     

血小板对ECV304细胞PTA1表达和PDGF释放的影响

目的:探讨血小板对妊高征患者血清刺激下的ECV304细胞(ECV304)表达人血小板/T细胞活化抗原1(PTA1)和释放血小板衍生生长因子(PDGF)的影响。方法①用间接免疫荧光染色结合流式细胞仪分析观察血小板与妊高征患者血清刺激的ECV304细胞孵育前后PTA1的表达②用ELISA方法检测血小板与妊高征患者血清刺激的ECV304细胞孵育前后PDGF的量。结果:①血小板与妊高征患者血清刺激(24h、48h)的ECV304细胞孵育后PTA1表达的百分率(6.32%、4.13%)明显低于对照组(15.51%、5.64%)(P<0.05)。②妊高征患者血清刺激ECV304细胞8h、24h、48h释放PDGF的量(1593、2625、2175ng/L)明显高于正常孕妇组(235、405、133.5ng/L)差异显著(P<0.01)。③血小板与妊高征患者血清刺激的ECV304细胞孵育后,24h、48h细胞培养上清液中PDGF-B的量又进一步增加(3266、2360ng/L),与对照组比差异显著(P<0.05)。结论:ECV304细胞与血小板之间的黏附可能是通过PTA1分子发挥作用的,同时导致了PDGF的进一步释放。     

驻极体诱导3T3细胞内游离钙离子浓度的变化

目的：研究公主体对3T3细胞内游离Ca^2 浓度变化的,体诱导3T3细胞凋亡与游离Ca^2 浓度变化之间的关系。方法：选用－300V和－500V驻极体作用3T3细胞48h,即时依次加入CaCl2,A23187,EDTA,通过流式细胞术检测细胞内Ca^2 浓度的变化和3T3细胞的凋亡量。结果：（1）负极性驻极体作用3T3细胞后,3T3细胞出现10％的凋亡量;（2）驻极体作用3T3细胞以后,细胞质游离Ca^2 浓度升高,Ca^2 浓度的升高不是由于细胞内钙池的释放,而是由于细胞外Ca^2 内流所引起,结论：驻极体诱导3T3细胞凋亡的机制可能是细胞外Ca^2 内上起的。     

卵巢癌细胞DNA倍体、p21及p53基因表达产物定量分析

目的　探讨卵巢癌DNA倍体、p2 1及 p5 3基因定量表达情况及其临床意义。 方法　应用流式细胞术 (FCM )和荧光免疫技术 ,对 3 9例卵巢癌组织的卵巢癌细胞和 9例正常卵巢组织的卵巢细胞的染色体倍体、p2 1基因和 p5 3基因表达产物进行定量分析。结果　在卵巢癌组织中DNA的异倍体率、p2 1、p5 3基因蛋白产物的阳性表达率均明显高于对照组 ,两者比较均有非常显著性差异 (P <0 .0 1)。结论　应用FCM定量测定卵巢癌细胞中的DNA倍体、p2 1及 p5 3基因表达产物 ,在临床上具有重要的参考价值     

Pathological analyses of early recurrence and malignant transformation in meningiomas

We investigated factors of the early recurrence and malignant transformation of histologically benign meningiomas using immunohistochemistry for MIB-1 positive indices (PI) and p53 protein expression, a flow cytometric DNA analysis, and the examination of numerical chromosomal aberrations detected by fluorescence in situ hybridization using an α-satellite DNA probe and abcr gene locus-specific probe. Twenty-six meningiomas of 23 patients were classified into two groups: the 3 patients in whom a recurrence was defined within two years after initial surgery and who showed histologically malignant features were classified as the early recurrent group, and the other 20 patients in whom recurrence did not develop during the same period were classified as the nonrecurrent group. DNA aneuploidy was observed in 40% of the nonrecurrent patients and in 67% of the early recurrent patients. Loss of chromosome 22 was the most common numerical aberration, but the aberrations characteristic of early recurrent meningiomas were not detected. The MIB-1 PI values of the early recurrent meningiomas were higher than those of nonrecurrent meningiomas, suggesting that MIB-1 PI is very important for biological and histopathological analyses and prediction of the future recurrence of meningiomas.