人外周血T淋巴细胞凋亡与胀亡研究及地塞米松的诱导作用

目的:观察分离培养的人外周血T淋巴细胞的调亡与胀亡.方法:密度梯度离心法及尼龙棉柱法分离健康成年人外周血T淋巴细胞,分空白组及地塞米松(DXM)组,培养后观察细胞光镜及电镜形态学、FDA/P1荧光染色,并用流式细胞仪检测凋亡和胀亡细胞比例变化.结果:①人外周血T淋巴细胞经体外培养,可自然出现典型的细胞凋亡与胀亡形态学改变.②人外周血T淋巴细胞在地塞米松诱导作用下,凋亡率与胀亡率皆有显著增高.③随着培养时间的增加人T淋巴细胞凋亡与胀亡率亦有升高.结论:人外周血T淋巴细胞存在凋亡与胀亡现象,DXM可诱导人外周血T淋巴细胞凋亡与胀亡.     

牙龈卟啉单胞菌诱导T淋巴细胞活化、凋亡的体外研究

目的检测牙龈卟啉单胞菌对人外周血中T淋巴细胞活化及凋亡的作用,并检测Fas/FasL在牙龈卟啉单胞菌(Porphyrom onas gingivalis,Pg)诱导的T淋巴细胞凋亡中的表达。方法选取10例全身及牙周组织健康受试者,分离外周血中T淋巴细胞,在有/无Pg情况下培养0~96 h,用荧光探针(Annexin V-FITC、PI、CD69)及特殊的单克隆抗体(Fas、FasL)进行标记,并进行流式细胞仪检测。结果 CD69+淋巴细胞+Pg组Annexin V+/PI-细胞百分数在各个时间点上都明显高于T淋巴细胞+Pg组(P0.01)。Fas和FasL的表达量明显上调。用抗Fas单克隆抗体阻滞Fas-FasL相互作用导致T细胞凋亡的明显减少,百分比为(20.56±2.43)%,未加抗体的为(50.41±2.68)%。但残余的细胞凋亡活动与阴性对照相比仍高。结论 Pg能够诱导人外周血中T淋巴细胞活化,并且能够通过活化促进其凋亡,Pg诱导T淋巴细胞凋亡主要通过Fas-FasL途径,并具有时间依赖性。     

氢化可的松诱导特发性血小板减少性紫癜淋巴细胞凋亡和增殖抑制

探讨氢化可的松对儿童急性特发性血小板减少性紫癜（AITP）外周血淋巴细胞增殖和凋亡的影响,对12例确诊的儿童AITP外周血淋巴细胞进行体外培养,用流式细胞术观察糖皮质激素处理后外周血淋巴细胞凋亡数量的变化,并用Wst-1（四氮唑盐）测定代表增殖的活细胞数。结果显示,激素处理组淋巴细胞凋亡率明显高于对照组,而活细胞数则明显低于对照组;激素处理后的外周血淋巴细胞凋亡率明显高于对照组,而活细胞数则明显低于对照组;激素处理后的外周血淋巴细胞凋亡率也明显高于处理前和正常对照组,均具有显性差异。结果表明,氢化可的松具有明显的诱导淋巴细胞凋亡和增殖抑制作用,说明糖皮质激素可通过诱导AITP外血淋巴细胞凋亡和增殖抑制发探治疗作用。     

TRAIL联合多西紫杉醇诱导喉鳞癌Hep-2 细胞凋亡的实验研究

目的：观察肿瘤坏死因子相关凋亡诱导配体( TRAIL) 联合多西紫杉醇应用于人喉鳞癌Hep-2 细胞生长的抑制增殖和诱导 凋亡作用。方法：实验分四组,1 组对照组,2 组为应用TRAIL组,3 组单独应用多西紫杉醇,4 组联合应用TRAIL及多西紫杉醇。 分别应用MTT、流式细胞仪检测细胞凋亡率,倒置显微镜观察细胞的形态学改变。结果：TRAIL 与多西紫杉醇联合作用于Hep-2 细胞,能显著增强对Hep-2 细胞的杀伤、抑制增殖及诱导凋亡作用,其联合应用的凋亡抑制率明显高于单独应用TRAIL组和多西 紫杉醇组( P<0.05)。结论：TRAIL与多西紫杉醇联用能显著提高对喉鳞癌Hep-2 细胞的生长抑制和诱导凋亡作用。     

TRAIL联合多西紫杉醇诱导喉鳞癌Hep-2细胞凋亡的实验研究

目的：观察肿瘤坏死因子相关凋亡诱导配体（TRAIL）联合多西紫杉醇应用于人喉鳞癌Hep-2细胞生长的抑制增殖和诱导凋亡作用。方法：实验分四组,1组对照组,2组为应用TRAIL组,3组单独应用多西紫杉醇,4组联合应用TRAIL及多西紫杉醇。分别应用MTT、流式细胞仪检测细胞凋亡率,倒置显微镜观察细胞的形态学改变。结果：TRAIL与多西紫杉醇联合作用于Hep-2细胞,能显著增强对Hep-2细胞的杀伤、抑制增殖及诱导凋亡作用,其联合应用的凋亡抑制率明显高于单独应用TRAIL组和多西紫杉醇组（P〈0．05）。结论：TRAIL与多西紫杉醇联用能显著提高对喉鳞癌Hep-2细胞的生长抑制和诱导凋亡作用。     

HIV-1 Tat蛋白通过增加线粒体ROS产生诱导人外周血B淋巴细胞凋亡

目的:探讨HIV-1 Tat蛋白对人外周血B淋巴细胞增殖、凋亡的影响及其机制。方法:采用流式细胞分选术分离HIV阳性患者外周血单核细胞的B淋巴细胞,分别转染pTat或pcDNA3.1各10μg(分别为pTat组与pcDNA3.1组),采用MTT实验检测细胞胞增殖情况,流式细胞术检测凋亡情况,DCHF-DA测定ROS水平,彗星试验检测细胞DNA损伤情况。结果:pTat组转染24h、48h的细胞增殖抑制率、细胞凋亡率及线粒体ROS水平均显著高于pcDNA3.1组(P0.05)。pcDNA3.1组细胞的DNA大部分呈圆形荧光团,无拖尾现象;pTat组的细胞DNA拖尾现象,呈现典型彗星图像。与pcDNA3.1组相比,pTat组细胞DNA尾长、尾部DNA比例均显著增加(P 0.05)。结论:HIV-1 Tat蛋白可能通过增加线粒体ROS产生,诱导DNA损伤,进而抑制人外周血B淋巴细胞增殖并促进其凋亡。     

地塞米松诱导培养的大鼠大脑皮质星形胶质细胞凋亡

目的　研究地塞米松诱导纯化培养的大鼠大脑皮质星形胶质细胞凋亡的作用。方法　不同浓度的地塞米松(浓度为 10 -3 、 10 -4、 10 -5mol/L)与纯化培养的大鼠大脑皮质星形胶质细胞共同孵育 18小时后 ,吖啶橙染色荧光显微镜观察细胞凋亡形态学改变 ,流式细胞仪检测细胞凋亡和结晶紫比色法酶标仪测定活细胞数。结果　 (1)吖啶橙染色荧光显微镜观察 :10 -4组偶见细胞凋亡 ,10 -3 组可见许多细胞有典型的凋亡形态学改变核固缩 ,深染 ,或肿胀 ,碎裂 ,并可见凋亡小体。 (2 )流式细胞仪检测细胞凋亡 :10 -3 组细胞凋亡率为 15 99% ,与其它三组相比明显增高 ,有显著性差异 (P <0 0 1)。 (3)结晶紫法酶标仪测定活细胞数 :10 -3 组OD值为 0 . 185与其它三组相比明显下降 ,有显著性差异 (P <0 0 1) ,表明活细胞数明显减少。结论　大剂量地塞米松可诱导体外的星形胶质细胞凋亡。     

罗仙子提取物对肝癌的体内外抑制作用研究

本文观察中药罗仙子提取物在离体细胞水平以及整体动物模型中对肝癌的治疗作用。在罗仙子提取物干预人肝癌SMMC-7721细胞24 h后,采用MTT法检测细胞活率;采用光学显微镜、荧光显微镜、透射电镜观察细胞形态学改变,流式细胞仪检测细胞凋亡率;以Heps肝癌小鼠模型观察罗仙子提取物体内对肿瘤生长的抑制作用。研究发现不同浓度罗仙子提取物可抑制SMMC-7721细胞增殖同时诱导典型的细胞凋亡形态学变化,与对照组相比细胞凋亡率显著升高,且具有浓度依赖性;不同浓度罗仙子提取物可显著抑制Heps肝癌小鼠体内肿瘤生长。实验证明罗仙子提取物可显著抑制Heps肝癌小鼠体内肿瘤生长,该作用可能与罗仙子提取物抑制肝癌细胞增殖、诱导细胞凋亡有关。     

甲壳胺诱导人肝癌HepG2细胞凋亡的实验研究

目的:通过体外实验探讨甲壳胺是否对人肝癌细胞HepG2具有生长抑制及诱导凋亡作用.方法:在HepG2细胞培养液中加入不同浓度的甲壳胺,培养48 h,于倒置相差显微镜下观察甲壳胺处理组及对照组细胞形态学变化;用流武细胞术(FCM)检测HepG2细胞的凋亡率,Western印迹检测甲壳胺处理组及对照组Bcl-2和p53蛋白...     

江浙蝮蛇蛇毒蛋白诱导K562细胞调亡的研究

目的：研究江浙蝮蛇蛇毒蛋白诱导K562细胞调亡。方法：通过电镜观察蛇毒蛋白作用后K562细胞的形态变化;MTT检测蛇毒蛋白对细胞增值的影响,同时应用流式细胞仪检测细胞凋亡数及其对细胞周期的影响;采用琼脂糖凝胶电泳观测凋亡片断。结果：蛇毒蛋白作用K562细胞后,能显著抑制细胞增值;LC50为4．96μg／mL,电镜可观察到凋亡形态学改变;电泳呈现典型的阶梯状条带,流式细胞仪检测到凋亡峰。结论：江浙蝮蛇蛇毒蛋白可诱导K562细胞调亡。     

白蚁与动物及微生物的共生类型及其机制

综述了白蚁螱客的主要种类、共生关系及相关机制的研究进展。白蚁螱客中,已报道的动物种类达170种。在与动物的共生关系中存在偏利共生（宾主共栖和异种共栖）、互利共生和无关共生三种;在与微生物的共生关系中,存在与内生菌（原生动物、细菌、真菌和放线菌）和外生菌（蚁巢伞菌等）间的互利关系。指出了白蚁与螱客研究中存在的问题,给出了解决方案,并提出了今后可能的研究热点或方向,为白蚁的综合利用（如纤维素酶）及今后研究物种间的协同进化提供了基础资料。     

Relative and combined effects of ethanol and protein deficiency on strontium and barium bone content and fecal and urinary excretion

To elucidate accumulation of minerals in human iliac arteries with aging, the content of minerals was analyzed by inductively coupled plasma atomic emission spectrometry. Bilateral common, internal, and external iliac arteries of 16 men and 8 women, ranging ages from 65 to 93 yr, were examined. It was found that an extremely high accumulation of calcium and phosphorus occurred in the common iliac artery at old age, being higher than that of the internal and external iliac arteries. It should be noted that the accumulation of calcium and phosphorus is the highest in the common iliac artery among the human arteries examined to date. Regarding sexual differences, the content of calcium and phosphorus in the common and internal iliac arteries was higher in women than in men, whereas their content in the external iliac artery was lower in women than in men.     

New amino-dithiaphospholanes and phosphoramidodithioites and their rhodium and iridium complexes

New sulfur derivatives of phosphoramidite ligands were synthesized and the impact of the sulfur unit on the spectroscopic properties of their rhodium and iridium complexes was investigated. The new ligands Bn₂NPSCH₂CH₂S^a(P-S^a) (Bn = benzyl, 4), Bn₂NPSCHCHS^a(CH₂)₃C^aH₂(P-S^a)(C^a-S^a) (6) and Bn₂NP(4-XC₆H₄OMe)₂ (X = S, 7a; X = O, 7b) were converted to the rhodium and iridium complexes trans-[Rh(CO)Cl(L)₂] (L = 4, 6, 7), [RhCl(COD)(L)] (L = 4, 6, 7), [IrCl(COD)(7a)] and [IrCl₂Cp∗(6)]. For comparison, some phosphoramidite complexes of these formulations also were synthesized. The new metal complexes were spectroscopically analyzed. For the carbonyl complexes, the ν_CO IR stretching frequencies were lower than for the corresponding phosphite and phosphoramidite ligands. The ¹J_PRh coupling constants for the rhodium complexes with the new ligands were also smaller than for the respective phosphoramidite and phosphite complexes. Finally, the ¹J_PSe coupling constants of the selenides of the new ligands were lower than those of the phosphoramidite ligands but higher than for PPh₃. The spectroscopic data reveal that the new thio ligands 4, 6 and 7a are more electron donating than phosphites and phosphoramidites but less electron donating than PPh₃.     

Astrocytes and Synaptosomes Transport and Metabolize Lactate and Acetate Differently

Astrocytes transport the monocarboxylate acetate, but synaptosomes do not. The reason for this is unknown, because both preparations express monocarboxylate transporters (MCT). The transport and metabolism of lactate, another monocarboxylate, was examined in these two preparations, and the results were compared to those for acetate. Lactate transport is more rapid in astrocytes than in synaptosomes, but of lower affinity (Kms of 17 and 4 mM, respectively). Lactate (0.2 mM) is metabolized to CO2 more rapidly in synaptosomes than in astrocytes (rates of 0.37 and 0.07 nmol x mg protein(-1) x min(-1), respectively). The reason for this is unclear, but cellular differences in lactate dehydrogenase isotype expression may be involved. Acetate is metabolized to CO2 more rapidly in astrocytes than in synaptosomes (rates of 0.43 and 0.02 nmol x mg protein(-1) x min(-1), respectively). This is likely due to cellular differences in the expression of monocarboxylate transporter subtypes.     

Rapporteur report: Basics and technology and metrology and standards

The first and second sessions of the Workshop focussed on the basics of ultrasound and infrasound, their applications in both industry and medicine, and metrology and protection standards for ultrasound applications.     

Prostaglandin and thromboxane production by human and guinea-pig macrophages and leucocytes

The ability of partially purified human and guinea-pig haematogenous cell populations, when cultured in vitro, to metabolise arachidonic acid (AA) has been studied. Supernatants from 24 hour cell culture have been subjected to analysis for products of AA metabolism by gas chromatography with electron-capture detection.The cell types studied were human peripheral blood monocytes (both glass adherent and non-adherent), neutrophils, eosinophils and leukemic leucocytes; thoracic duct lymphocytes and lung alveolar macrophages. From the guinea-pig, induced and non-induced macrophage or neutrophil enriched peritoneal exudate populations, lymph node cells, peritoneal eosinophils and peripheral blood platelets were examined. Supernatants were assayed for the presence of PGE₂, PGD₂, PGF_2α, TXB₂ and 6-keto-PGF_1α. In all types studied PGE₂ and TXB₂ were the major products formed. The identification of PGE₂ and TXB₂ was confirmed by GC/MS with multiple ion monitoring.The results have been compared with other reports and their possible significance discussed in relation to the proposed role of prostaglandins as mediators and modulators in immunopathology.     

Cytoskeleton and mitochondrial morphology and function

It has been well established that the cytoskeleton is an essential modulator of cell morphology and motility, intracytoplasmic transport and mitosis, however cytoskeletal linkage to the organelles has not been unequivocally demonstrated. Indeed, cytoskeleton appears to be essential in determining and modulating gene phenotype as a function of cellular environment. According to recent studies, the organization of the cytoskeleton network together with associated protein(s) could be essential in regulating mitochondrial function and particularly the permeability of the mitochondrial outer membrane to ADP. The aim of this chapter is to summarize the main properties of the cytoskeletal environment of mitochondria and the possible role(s) of this network in mitochondrial function in myocytes.     

Cross-reactivity between storage and dust mites and between mites and shrimp

Many patients have sensitivities to multiple species of storage and house dust mites. It is not clear if this is because patients have multiple sensitivities to species-specific mite allergens or if these mites share many cross-reacting allergens. Our objective was to further define the cross-allergenicity between several species of storage and house dust mites using crossed-immunoelectrophoresis (CIE), crossed-radioimmunoelectrophoresis (CRIE), immunoblotting, and ELISA. CIE and CRIE reactions revealed that storage mites shared two cross-antigenic molecules and one of these bound IgE in a serum pool from mite allergic patients. Antibody in anti-sera built to each species of mite recognized many SDS–PAGE resolved proteins of other mite species and this suggested the potential for other cross-reactive allergens. Among patient sera, IgE bound to many different proteins but few had IgE that bound to a protein with common molecular weights across the mite species and this suggested mostly species-specific allergens. Antiserum built to each mite species precipitated one protein in shrimp extracts that bound anti-Der p 10 (tropomyosin) and IgE in the serum pool. Anti-Der p 10 showed strong binding to shrimp tropomyosin but very little to any of the mite proteins. ELISA showed the mite extracts contained very little tropomyosin. The storage and dust mites investigated contain mostly species-specific allergens and very small amounts of the pan-allergen tropomyosin compared to shrimp and snail.