谷胱甘肽保护的金纳米团簇对HeLa细胞毒性研究

目的 探究由谷胱甘肽作为表面保护剂的金纳米团簇(GSH-Au NCs)对宫颈癌HeLa细胞株的毒性影响.方法 利用荧光分光光度计测定用含有GSH-Au NCs的培养基处理HeLa细胞后不同时间点的荧光强度,观察HeLa细胞对GSH-Au NCs在1、2、6、12、24 h内的摄取情况.同时采用BALB/c荷瘤小鼠进行体内实验,分别腹腔注射0.2 ml浓度为3 mmol/L的GSH-Au NCs和等体积的蒸馏水(对照组)后24 h取出肿瘤组织,通过电感耦合等离子体质谱(ICP-MS)检测组织中的金元素含量以探究纳米团簇在肿瘤处随时间的摄取情况.最后用噻唑蓝(MTT)比色法研究不同浓度(0.003～0.3 mmol/L)的GSH-Au NCs处理HeLa细胞24、48 h的细胞毒性.结果 HeLa细胞对GSH-Au NCs的摄取率在24 h内不断升高,24h时达峰值73.13％.荷瘤小鼠实验表明,腹腔注射GSH-Au NCs 24 h后,肿瘤组织对GSH-Au NCs的摄取量(320±15) ng/g较对照组(腹腔注射蒸馏水)高,差异具有统计学意义(P＜0.05).用不同浓度的GSH-Au NCs处理HeLa细胞24 h,对细胞存活率有轻微影响,随浓度升高对细胞的抑制作用更为明显,GSH-Au NCs浓度为0.3 mmol/L时的HeLa细胞存活率降为对照组(GSH-Au NCs浓度为0)的86％(P＜0.05);处理48 h时,各浓度组的细胞存活率与对照组间差异均无统计学意义(P＞0.05).结论 虽然GSH-Au NCs在体外和体内均易被细胞和肿瘤组织摄取,但其本身对HeLa细胞并无明显细胞毒性,可安全应用于影像、载药及靶向给药等生物医药领域.     

性别对L-精氨酸诱发的昆明小鼠慢性胰腺炎的影响

目的:通过比较雌雄小鼠L-精氨酸诱发的慢性胰腺炎(CP)程度的差异,探讨性别对CP模型形成的影响。方法:健康昆明小鼠雌雄各42只,分为雌性对照组、雌性CP组、雄性对照组和雄性CP组(对照组n=18,每时点6只;CP组n=24,每时点8只)。CP模型组小鼠腹腔注射20%L-精氨酸(3 g/kg,每周1次,每次2轮,间隔1 h)诱发CP。分别于造模后第2、4、6周处死动物,取小鼠胰腺组织,HE及Masson染色检测各组小鼠胰腺形态学改变及纤维化程度;免疫组织化学观察胰腺组织F4/80的阳性染色率;real-time PCR检测胰腺组织白细胞介素6(interleukin-6,IL-6)、α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)及纤维连接蛋白(fibronectin,FN)的mRNA表达;Western blot检测胰腺组织α-SMA及FN蛋白的表达。结果:20%L-精氨酸腹腔注射后的第2、4和6周,HE及Masson染色显示雌雄CP组胰腺组织均可见病理损伤,但是同一时点雌雄小鼠胰腺损伤程度存在明显差异,雄性小鼠胰腺病变程度明显较重;胰腺F4/80染色显示雄性小鼠胰腺F4/80的表达水平在造模后各时点均明显高于同一时点的雌性小鼠;造模后第2和4周胰腺IL-6的mRNA表达在雌雄CP组均有所升高,但是雄性组表达水平明显高于雌性组(P0.05)。造模后α-SMA和FN在mRNA水平和蛋白质水平均可见高表达,但雄性小鼠表达时点更早,水平更高(P0.05)。结论:采用腹腔注射20%L-精氨酸可成功复制小鼠CP模型,但复制的模型在雌雄小鼠存在病变程度差异。L-精氨酸诱导的CP模型在雄性小鼠成模更早、纤维化程度更明显,其原因可能与雄性小鼠对L-精氨酸更敏感,引发的炎症反应程度更明显有关。雄性小鼠更适合于复制CP动物模型。     

不同性别肾缺血再灌注损伤模型小鼠MicroRNA的表达谱

背景：有研究表明肾缺血再灌注损伤存在性别差异,但其具体机制有待进一步研究。 目的：观察雌雄性小鼠缺血前后肾脏组织中MicroRNA表达及其差异,以期进一步研究MicroRNA在肾缺血再灌注损伤性别差异中的作用及机制。 方法：将雄性和雌性小鼠分别肾缺血45 min再灌注损伤24 h,同时设置雄性和雌性假手术组作对照。采用MicroRNA基因芯片技术检测雌雄小鼠肾缺血45 min再灌注损伤24 h及假手术后肾组织MicroRNA表达的差异,样品间表达差异的阈值为2倍。 结果与结论：在雌性肾缺血再灌注与雄性肾缺血再灌注组对比发现有表达上调的MicroRNA有5个;雌性肾缺血再灌注组与雌性假手术组对比发现有差异表达的MicroRNA有29个,其中上调的有MicroRNA有25个,下调的MicroRNA有4个;雄性肾缺血再灌注组与雄性假手术组对比发现有差异表达的MicroRNA有38个,其中上调的MicroRNA有9个,下调的MicroRNA有29个;雌性假手术组与雄性假手术组对比发现有差异表达的MicroRNA有102个,其中上调MicroRNA的有22个,下调的MicroRNA有80个。结果说明,不同性别小鼠肾缺血再灌注前后肾组织中微小核糖核酸表达存在差异,这些MicroRNA的差异表达可能导致不同性别小鼠肾脏对缺血再灌注损伤的敏感性及耐受性存在差异。 中国组织工程研究杂志出版内容重点：肾移植;肝移植;移植;心脏移植;组织移植;皮肤移植;皮瓣移植;血管移植;器官移植;组织工程全文链接：     

2.128大鼠性别及动情周期对内脏感觉影响的研究

目的研究生理状态下大鼠内脏感觉的性别差异,及雌性大鼠动情周期对内脏感觉的影响. 方法选取清洁级成年SD大鼠,雌雄各8只.根据阴道涂片,将雌性大鼠分为两期:动情前期和动情期归为A组(体内雌激素高水平组),动情后期和动情间期归为B组(体内雌激素低水平组),雄性大鼠为C组.采用结肠直肠气囊注气扩张法,分别测定A、B、C三组大鼠的行为反射,重复2个动情周期.     

四氯化碳致小鼠急性肝损伤的性别差异及其与雄激素受体表达的关系

目的比较CCl4对不同性别小鼠急性肝损伤的诱导作用及其对肝脏雄激素受体(AR)表达的影响。方法健康成年昆明小鼠70只,随机分为CCl4雄性组、CCl4雌性组、雄性对照组,雌性对照组。50%CCl4-粟米油溶液皮下注射,0.6ml/100g体重,1周2次,诱导小鼠急性肝损伤。造模第7天处死各组动物取材固定,常规HE染色比较不同组别小鼠肝组织病理变化;免疫组织化学法检测各组小鼠肝脏AR的表达,根据阳性细胞反应强度及百分比设定0级(阴性)及1-3级(阳性)标准,双盲计分法统计各组AR表达分值。结果实验组CCl4造模过程中雄性组小鼠死亡率为60%(15/25),明显高于雌性组(12%,3/25),对照组小鼠无死亡(0/10,0/10)。肝组织HE染色结果可见CCl4诱导的急性肝损伤主要部位为肝小叶周围的门管区及界板区,损伤部位肝细胞出现脂肪变,少数肝组织出现点状坏死。免疫组化显示雄性及雌性对照组小鼠肝细胞AR呈弱阳性表达,不同性别小鼠肝细胞AR表达差别无统计学意义;实验组各组别肝小叶周围受损肝细胞出现较强的AR表达,其中CCl4雄性组AR表达分值明显高于CCl4雌性组小鼠(P<0.01)。结论雄性小鼠对急性肝损伤的耐受性低于雌性小鼠,其中AR的高表达与CCl4诱导的急性肝损伤性别差异密切相关。     

蜜炼川贝枇杷膏灌胃给药SD大鼠14天毒性探索研究

目的 研究京都念慈庵蜜炼川贝枇杷膏可能的毒性反应,为临床或家庭用药安全性提供参考。方法 采用大鼠14 d重复给药毒性试验,SD大鼠,按体重随机分成对照组、枇杷膏低、中、高剂量组,每组10只,雌雄各半,共40只,给药剂量分别为0、2.5、5、10 ml/（kg·d）,给药体积为10 ml/（kg·d）,相当于人临床用量的2.5、5、10倍。每天经口灌胃给药1次,连续14 d。第15天测定血液学、血液生化学各指标,同时测定心、肝、脾、肾脏器重量并计算脏器系数。结果 雌雄大鼠各剂量组体重正常增长,与对照组比较,差异无统计学意义（P＞0.05）。雄性大鼠各给药组血液学检测指标与对照组比较,差异无统计学意义（P＞0.05）;雌性大鼠中剂量组MCV偏低,与对照组比较,统计学意义显著（P＜0.01）;雄性大鼠中、高剂量组BUN降低,与对照组比较,统计学意义显著（P＜0.01）;雌性大鼠各给药组血液生化学检测指标与对照组比较,差异无统计学意义（P＞0.05）。雌性、雄性大鼠的脏体比数据中,中剂量组肝系数升高,与对照组比较,差异具有统计学意义（P＜0.05）。结论 蜜炼川贝枇杷膏SD大鼠重复给药14d未提示有明显毒性反应。     

小鼠子宫肥大细胞在给天花粉后定量、组化,电镜的研究

小鼠子宫肥大细胞在给天花粉后定量、组化，电镜的研究龚凤英杨美林（山西医科大学组胚教研组）中国１号雌性小鼠３６只，分为实验组和对照组，均为动情期。对照组不作任何处理，实验组分别腹腔注射天花粉，于注药后２０ｈ，２１ｈ，２２ｈ，２４ｈ处死，取子宫组织，作光...     

雌激素受体β在成年C57小鼠脑内的表达与性别差异

目的:观察雌激素受体β(ER-β)在成年小鼠脑内的性别差异.方法:成年C57小鼠用硫酸镍铵增强显色的免疫组织化学SP法.结果:ER-β在脑内有广泛的分布,ER-β免疫阳性产物主要表达于细胞核,但在个别脑区也可在胞质甚至突起内检测到.ER-β的表达具有一定的性别差异,在部分脑区ER-β的表达表现为雄性强于雌性,而在另一些脑区则相反,并且该受体的脑核团定位方面表现出某些性别差异.结论:在不同性别的动物同一核团,ER-β发挥的调节作用可能不一样,这可能是造成脑的性别差异的原因之一.     

雌性小鼠骨髓移植给雄性小鼠后内源性骨髓细胞残存状态的研究

 目的:以雌性小鼠骨髓移植给雄性小鼠的方法,通过检测雄性小鼠血细胞的Y染色体来明确内源性骨髓细胞的残存状态。方法:将雌性或雄性C57BL/6小鼠作为受体,实验组用［137Cs］照射,6 h后每只经尾静脉注射供体小鼠骨髓细胞1×107。统计骨髓移植后14 d动物的存活率,并通过眶静脉采血观察外周血白细胞数量的变化,检测受体雄性小鼠体内Y染色体基因水平的变化以明确骨髓移植的效果。结果:分别用1 000、950和900 rad的照射剂量对受体小鼠进行照射后将供体小鼠的骨髓移植到受体小鼠体内,1 000和950 rad剂量时雌性受体小鼠可迅速恢复造血功能,而雄性受体小鼠则仅有48%的存活率。900 rad照射剂量骨髓移植后,雄性受体小鼠迅速恢复了造血功能,13 d后外周血白细胞计数基本恢复正常。移植后的雄性受体小鼠在5周内已检测不到外周血细胞Y染色体基因,表明雄性受体小鼠的骨髓被完全破坏,雌性供体小鼠的骨髓可完全替代受体雄性小鼠的骨髓并且发挥造血功能。结论: 在照射剂量900 rad照射后,雄性小鼠可以作为骨髓移植受体,为将来应用雄性小鼠作为骨髓移植受体动物开展有关心血管疾病的研究奠定了实验基础。     

恩替卡韦抑制乙型肝炎病毒复制型小鼠体内病毒的复制

目的 了解乙型肝炎病毒（HBV）复制型小鼠模型能否用于抗HBV药物的筛选。方法 采用高压注射体内转染法将HBV复制型重组质粒pHBV4.1导入雄性BALB/C小鼠,转染后24h按体重、年龄、血清HBeAg水平对小鼠进行配对,分为实验组和对照组（n=4）,2组小鼠分别连续3d（每日1次）给恩替卡韦和生理盐水灌胃。最后1次给药后24h处死小鼠。分别用Northern、Southern杂交检测小鼠肝脏HBV mRNA和HBV DNA复制中间体;ELISA检测血清中HBeAg和HBsAg。结果 重复2次实验的结果显示,实验组小鼠HBV DNA复制中间体水平明显低于对照组,而两组小鼠HBV mRNA、HBeAg和HBsAg水平无明显差异。结论 恩替卡韦抑制HBV复制型小鼠体内HBV的复制,提示该小鼠模型可以用于抗HBV药物的初步筛选。     

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.     

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.     

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.     

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.