抗Fas的抗体在实验性病毒性心肌炎中作用机制的研究

目的探讨中和型抗Fas的抗体对小鼠柯萨奇病毒性心肌炎的治疗作用及作用机制。方法60只BAIB/c小鼠随机分为4组：1组空白对照组，2组病毒对照组、3组IgG对照组、4组抗Fas抗体治疗组。于接种后第10天每组随机处死小鼠8只，心肌组织切片HE染色了解心肌损伤情况，电镜技术及原位末端标记法（TUNEL）检测心肌细胞凋亡情况，免疫组化方法检测casimse-3的表达，逆转录．聚合酶链反应（RT-PCR）检测心肌组织中easpase-3的mRNA表达。实时定量PCR（RQ-PCR）定量心肌柯萨奇病毒B3（CVB3）mRNA拷贝数。结果（1）病毒对照组可见caspase-3表达和心肌细胞凋亡，且均与心肌病变积分成正相关（r=0．81，P〈0．01；r=0．73，P〈0．05）。（2）抗Fas抗体治疗组小鼠心肌组织病变积分、心肌细胞凋亡率、caspase-3蛋白和mRNA表达及心肌内CVB3 mRNA拷贝数均明显低于病毒对照组（P〈0．05）和IgG对照组（P〈0．05）。结论Fas/FasL途径介导的心肌细胞凋亡参与了病毒性心肌炎的发病过程，凋亡是导致病毒性心肌炎心肌损伤的重要机制之一，抗Fas抗体可降低caspase-3活化和mRNA表达，减少心肌细胞凋亡，降低心肌细胞内病毒复制，使心肌损伤明显减轻。     

先天性心脏病肺动脉高压对左室心肌细胞凋亡的影响及与心肌AngⅡ含量的相关分析

目的探讨先天性心脏病（CHD）合并肺动脉高压（PAH）对左室心肌细胞凋亡的影响以及与心肌局部血管紧张素Ⅱ（angiotensinⅡ，AngⅡ）含量变化的相关关系。方法采用心肌细胞原位末端标记、左室心肌凋亡基因免疫组织化学染色及半定量分析和HPLC．RIA法分别检测了2001年10月至2003年2月因先天性心脏病、先天性心脏病合并肺动脉高压（PASP〉30mmHg，PAMP〉20mmHg）死亡的14例患儿心肌细胞凋亡和5种相关凋亡基因蛋白表达及心肌AngⅡ的含量。结果与对照组相比较，CHD组的左室心肌细胞凋亡指数和凋亡基因的蛋白表达明显高于对照组（P〈0．01），而CHD合并PAH组又高于CHD组（P〈0．05），单纯CHD及CHD合并PAH患儿心肌组织AngⅡ均显著高于对照组，CHD合并PAH患儿高于单纯CHD，并且CHD合并PAH患儿组织AngⅡ与心肌细胞凋亡指数和凋亡基因的表达量呈显著相关（相关系数分别为0．522，P〈0．01，0．546，P〈0．05），而CHD合并PAH患儿PASP与心肌细胞凋亡指数亦呈正相关（相关系数分别为0．591，P〈0．05）。结论肺动脉压和多种因素直接和或间接引起的组织AngⅡ升高在CHD合并PAH左室心肌细胞凋亡的过程中共同发挥了直接和或间接的作用，为PAH对左心功能的影响提供了实验依据。     

红细胞生成素对大鼠阿霉素性心肌病的预防

目的 探讨红细胞生成素（EPO）对阿霉素（DOX）性心肌病的预防性保护作用及可能机制.方法 31只Wistar大鼠随机分为DOX组（12只）、DOX＋EPO组（11只）和对照组（8只）,前2组采用腹腔注射DOX建立扩张型心肌病模型,并在腹腔注射DOX前分别予生理盐水或EPO预防性干预.测量3组大鼠血流动力学以了解左室收缩功能变化;Masson氏染色法观察心脏组织病理学变化;TUNEL法分析心肌细胞的凋亡;Western印迹法检测Bax和Bcl-2的蛋白表达.结果 DOX组和DOX＋EPO组左室收缩功能差异有统计学意义（P〈0.05）,DOX＋EPO组的心肌纤维化面积比率（7.49±1.11）%明显比DOX组（12.14±1.07）%降低（P〈0.05）.DOX组和DOX＋EPO组凋亡指数分别为（0.93±0.08）%和（0.16＋0.04）%（P〈0.05）.Western印迹显示,DOX组中Bcl-2蛋白表达水平低于对照组,DOX＋EPO组Bcl-2的表达水平明显高于DOX组,差异均有统计学意义（P〈0.05）.结论 EPO可能通过上调Bcl-2蛋白表达发挥抗凋亡作用,从而对阿霉素性心肌病发挥预防性保护作用.     

法洛四联征右室流出道肥厚心肌细胞凋亡、Bax及Bcl-2蛋白表达情况

目的研究法洛四联征右室流出道肥厚心肌细胞凋亡、Bax及Bcl-2蛋白表达情况，并探讨其对手术时机的选择的意义。方法36例法洛四联征按年龄分成三个组。A组：≤2岁组；B组：〉2岁，且≤10岁；C组：〉10岁。用末端标记原位细胞凋亡法和免疫组化法检测其右室流出道肥厚心肌细胞凋亡指数，Bax及Bcl-2蛋白表达水平。结果心肌细胞凋亡指数，A组与B组和C组相比较（P〈0．05），A组最小。随年龄增长，Bax表达明显增多（P〈0．05），Bcl-2表达先增后减，但是Bcl-2／Bax比值随年龄增长而逐渐减少（P〈0．05）。结论不同年龄阶段法洛四联征右室流出道肥厚心肌细胞凋亡指数、Bax、Bcl-2蛋白表达存在明显差异。2岁内进行根治手术，预后较好。     

苯甲酸雌二醇致雌性大鼠胸腺退化作用的研究

目的探讨苯甲酸雌二醇对大鼠胸腺bcl-2和bax蛋白表达的影响。方法雌性大鼠行卵巢切除后给以苯甲酸雌二醇皮下注射，分别于注射后1、4、7天取胸腺，计算胸腺指数，用半薄切片甲苯胺蓝染色检测胸腺细胞凋亡情况，免疫组织化学法检测胸腺组织中bcl-2和bax蛋白的表达情况，统计学处理采用t检验。结果卵巢切除组大鼠胸腺指数明显增加，给雌二醇组大鼠胸腺指数明显减小（P〈0．05）；雌二醇处理组大鼠胸腺中可发现凋亡细胞和凋亡小体；大鼠胸腺中bcl-2蛋白表达量均低于对照组，（P〈0．001），而bax蛋白表达量均高于对照组，（P〈0．001）。结论苯甲酸雌二醇可抑制胸腺组织中bcl-2蛋白的表达，促进bax蛋白的表达，促进雌性大鼠胸腺退化，诱导大鼠胸腺细胞凋亡。     

食管癌组织中KAI1、EphA2蛋白的表达及其意义

目的探讨KAI1蛋白和EphA2蛋白的表达与食管癌发生及浸润转移的关系以及两者表达的相关性。方法采用免疫组化方法,检测160例食管癌组织及其150例正常黏膜中KAI1和EphA2蛋白的表达。结果KAI1蛋白在癌组织中的表达明显低于其正常黏膜（P〈0．05）,在有淋巴结转移病例中的表达明显低于无淋巴结转移病例（P〈0．05）,KAI1蛋白的表达与肿瘤的分化程度、浸润深度无关（P〉0．05）。EphA2蛋白在食管癌组织中的表达明显高于其正常黏膜（P〈0．05）,在深层浸润组的表达明显高于浅层浸润组（P〈0．05）,在有淋巴结转移组中的表达明显高于无淋巴结转移组（P〈0．05）,EphA2蛋白的表达与肿瘤的分化程度无关（P〉0．05）。KAI1蛋白和EphA2蛋白的表达在有淋巴结转移的病例中有显著负相关（P〈0．05）。结论 KAI1蛋白的低表达和EphA2蛋白的高表达与食管癌的癌变及转移的发生有关,两者对于食管癌的淋巴结转移具有负向调节作用。     

氯化镧对大鼠生长素分泌调节影响的体外研究

目的：研究氯化湖（LaCl3）对大鼠生长素分泌调节轴的影响。方法：采用荧光法检测离体脑片培育液中单胺类神经递质含量;用放射免疫法测垂体脑片育液中生长素（GH）含量。结果：①0．01mmol／LLaCl3垂体组育液中GH含量明显高于对照组（P＜0．05）;而1mmol／LLaCl3垂体组有液中GH含量明显低于对照组（P＜0．05）。②0.01mmol／LLaCl3下丘脑＋垂体组,其有液中GH含量则明显高于对照组（P＜0．01）和同剂量的单纯垂体组（P＜0．01）;但1mmol／L下丘脑＋垂体组育液中GH含量与对照组比较无显著差异（P＞0．05）。③0．01mmol／LLaCl3下丘脑组有液中NE含量明显高于对照组（P＜0．01）,5－HT明显低于对照组（P＜0.05）。④1mmol／L下丘脑组有液中NE和DA含量明显少于对照组（P＜0．01和0．05）,而5－HT含量则明显高于对照组（P＜0．001）。结论：低剂量氯化调可通过下丘脑和腺垂体双重作用促进GH分泌。     

转染Akt-1基因对大鼠心肌缺血-再灌注损伤的保护作用研究

目的通过观察转染Akt-1基因对大鼠心肌缺血-再灌注（I-R）损伤心脏功能及细胞凋亡相关蛋白的影响,探讨Akt-1基因对心肌I-R损伤保护作用的机制。方法使用结扎／松解左冠状动脉前降支法,结扎冠状动脉前降支30min,再灌注2h,建立大鼠在体心肌I-R模型。40只雄性SD大鼠随机分为五组：假手术组（S组）,缺血-再灌注组（I—R组）,转染Akt-1基因组（Gene组）,空载体组（VC组）,抑制剂组（AB组）,每组8只。Gene组术前48h开胸心肌内直接分点注射脂质体Akt-1质粒复合物,S组和I-R组分别注射同等体积PBS,VC组注射等体积脂质体,AB组注射等体积脂质体Akt-1质粒LY294002混合物。所有大鼠经颈动脉插管至左心室记录HR、LVSP、LVEDP和±do／dtmax变化,TTC染色法测定心肌梗死范围,TUNEL法原位标记凋亡心肌细胞,Western Blot测定Akt-1、Casepase-3蛋白表达,免疫组化检测Bcl-2、Bax表达。结果1．Gene组较I—R组、VC组和AB组左室心功能（HR、LVSP、LVEDP、±dp／dtmax）改善（P〈0．05）。2．Gene组凋亡指数（AI）及心肌梗死范围较I-R组、VC组及AB组均显著减少（P〈0．05）,但仍高于S组（P〈0．05）;S组细胞凋亡阳性细胞几无表达,心肌细胞凋亡指数（AI）明显低于其余各组（P〈0．05）。3．各组均可见Akt-1、Casepase-3蛋白表达,但S组中蛋白较其余各组减少（P〈0．05）;Gene组Akt-1蛋白表达较I-R组、VC组及AB组均增加（P〈0．05）;Gene组Casepase-3蛋白表达则较I-R组、VC组及AB组减少。4．S组Bcl-2和Bax表达较其余各组减少（P〈0．05）;Gene组较I-R组、VC组及AB组Bcl-2表达上调而Bax表达下调（P〈0．05）。结论Akt-1基因转染对I—R心肌具有保护作用,该作用可能与促进Bcl-2表达,抑制Bax和Casepase-3表达从而抑制凋亡的发生有关。     

慢性乙型肝炎患者外周血淋巴细胞CD8+CD38+的检测意义

目的：通过测定慢性乙型肝炎患者外周血淋巴细胞CD8^＋CD38^＋的表达及门冬氨酸氨基转移酶（AST）、总胆红素（TBIL）、凝血酶原时间（PT），旨在为治疗慢性乙型肝炎提供有用的参考指标。方法：流式细胞术检测38例慢性乙型肝炎、14例肝硬化患者外周血淋巴细胞CD8^＋CD38^＋细胞的百分率；同时测定AST、TBIL和PT。结果：（1）慢性乙型肝炎组CD8^＋CD38^＋、CD38^＋细胞均明显高于正常组（P〈0．01，P〈0．01），肝硬化组CD8^＋CD38^＋、CD38^＋细胞均明显高于正常组（P〈0．05，P〈0．05）。肝硬化组CD8^＋CD38^＋、CD8^＋细胞均明显低于慢性乙型肝炎组（P〈0．05，P〈0．05）。（2）慢性乙型肝炎轻、中、重各组之间比较，中度组CD8^＋CD38^＋高于轻度组（P〈0．05），重度组CD8^＋CD38^＋、CD8^＋、CD38^＋均高于轻度组（P〈0．05，P〈0．05，P〈0．05）；重度组CD38^＋高于中度组（P〈0．05）；肝硬化组CD8^＋CD38‘均明显低于轻、中、重各组（P〈0．05，P〈0．01，P〈0．01），肝硬化组CD8^＋低于重度组（P〈0．01）。（3）慢性乙型肝炎患者AST、TBIL、PT不正常组CD8^＋CD38^＋均高于AST、TBIL、PT正常组。结论：慢性乙型肝炎患者CD8^＋CD38^＋细胞明显升高，表明慢性乙型肝炎患者细胞免疫处于异常激活状态，并与肝功能损伤有一定的相关性。慢性乙型肝炎患者外周血淋巴细胞CD8^＋CD38^＋的测定，对病情分析和诊断有一定的临床参考价值。     

细胞增殖和凋亡失调在肺曲菌病发病中的意义

目的 观察细胞增殖与凋亡在肺曲菌病中的表达特征,探讨其在发病中的意义。方法 选用成人正常肺组织10例(对照组1)、肺曲菌病周围相对正常肺组织20例（对照组2）及肺曲菌病20例(病例组),采用免疫组化法检测KI-67、BCL-2和BAX的表达。结果 肺曲菌病病变区支气管上皮细胞KI-67和BAX的表达明显强于两对照组（P<0.01）;BCL-2的表达明显弱于两对照组（P<0.05）;BAX/BCL-2的比值明显高于两对照组（P<0.01）;肺曲菌病3种亚型中,BAX/BCL-2的比值在IPA中显著高于ABPA和曲菌球（P<0.05）。结论 细胞增殖和凋亡的失调在肺曲菌病发生发展中起重要的作用。     

Development in in vitro toxicology

There are three principal pressures driving the development of in vitro toxicology: (1) the need for more efficient testing systems to cope with the large number of xenobiotics currently being developed; (2) public pressure to reduce animal experimentation; and (3) a need for a better understanding of the mechanisms of toxicity. Within this, in vitro toxicology is focused on local, systemic, and target-organ toxicity. It is becoming increasingly apparent that a step or decision-tree approach using input of a variety of experimental data (physicochemical properties, biokinetics, cytotoxicity) provides the most efficient system for predicting toxicity. Examples of the use of in vitro toxicity systems for prediction of systemic toxicity and target-organ (liver) toxicity are presented.Originally presented at ECCP 93.     

Seasonal variations in acute toxoplasmosis in pregnant women in Slovenia

Between December 1999 and December 2004, 40 081 pregnant women were examined for toxoplasmosis with Toxo-IgG, Toxo-IgM enzyme immunoassay. Women with positive results were then retested with the Toxo-IgG avidity assay for recent toxoplasmosis. Recent acute toxoplasmosis in pregnant women was found to be significantly more frequent (p < 0.01) during winter than summer. The incidence of acute toxoplasmosis during winter-spring was also significantly more frequent (p < 0.025) than summer-autumn. This phenomenon should be taken into account when formulating preventive measures for toxoplasmosis, especially for pregnant women.     

Age-related changes in polyamines in memory-associated brain structures in rats

Polyamines putrescine, spermidine and spermine are positively charged aliphatic amines and have important roles in maintaining normal cellular function, regulating neurotransmitter receptors and modulating learning and memory. Recent evidence suggests a role of putrescine in hippocampal neurogenesis, that is significantly impaired during aging. The present study measured the polyamine levels in memory-related brain structures in 24- (aged), 12- (middle-aged) and 4- (young) month-old rats using liquid chromatography/mass spectrometry and high performance liquid chromatography. In the hippocampus, the putrescine levels were significantly decreased in the CA1 and dentate gyrus, and increased in the CA2/3 with age. Significant age-related increases in the spermidine levels were found in the CA1 and CA2/3. There was no difference between groups in spermine in any sub-regions examined. In the parahippocampal region, increased putrescine level with age was observed in the entorhinal cortex, and age did not alter the spermidine levels. The spermine level was significantly decreased in the perirhinal cortex and increased in the postrhinal cortex with age. In the prefrontal cortex, there was age-related decrease in putrescine, and the spermidine and spermine levels were significantly increased with age. This study, for the first time, demonstrates age-related region-specific changes in polyamines in memory-associated structures, suggesting that polyamine system dysfunction may potentially contribute to aged-related impairments in hippocampal neurogenesis and learning and memory.     

休克过程中肾上腺髓质素变化的研究进展

Adrenomedullin (AM) is a new peptidergic regulator of vascular function. AM serves as a hormone, which has many biological properties, plays an important role in the many pathophysiological processes, especially shock. This review will highlight the structure, biological properties of AM and the relationship between AM and shock.     

Secular change in menarche in women in Madrid

The age at menarche was estimated by recollection in 1617 women between the ages of 18 and 60 in Madrid and a nearby suburb, Pinto. The population of Pinto is working-class and the Madrid group, taken from residential neighbourhoods , belongs to the upper middle class. In both groups we found a diminution in average age at menarche, from 14.04 to 13.02 years in Madrid and from 14.55 to 13.16 years from about 1935 to about 1965 in Pinto. These changes have been more intense in the group which is less well-off economically, where living conditions have varied much more drastically.     

Uteroglobin in the developing rabbit conceptus in vivo and in vitro

Summary Uteroglobin (UGL) was measured in day- 4 to day-10 rabbit conceptuses by a competitive ELISA. Levels in blastocyst fluid, tissues, coverings and in the early fetus were determined separately. The total amount of UGL increased from 18.4 ng to 6.8 g per conceptus. The UGL content of individual day-6 blastocysts was studied in vitro. Culturing was carried out up to 60 h in Ham's F10 medium with polyvinylpyrrolidone as macromolecular component, with and without progesterone, and with progesterone plus estradiol. UGL was determined in the blastocyst fluids, tissues with coverings and in the culture media. After labelling with [³⁵S]-methionine, protein patterns of total blastocysts and of culture media were analysed by two-dimensional gel electrophoresis and fluorography. The morphology of cultured blastocysts was examined by electron microscopy. During 60 h of culture, the blastocysts expanded in diameter by 84%, and released 19% of their initial UGL content into the medium, independent of the hormonal substitution. Neither de novo synthesis, nor degradation of UGL was found: the protein remained unlabelled in fluorography, and its total quantity was not significantly different from that of non-cultured controls. Trophoblast, endoderm and embryoblast cells showed well preserved cell organelles and intercellular junctions, while the morphological differentiation of the germ layer was inhibited.