EMMPRIN在大鼠肾间质纤维化过程中的表达变化

目的 检测细胞外基质金属蛋白酶诱导因子(EMMPRIN)在大鼠肾间质纤维化模型中的表达变化.方法 64只Sprague-Dawley大鼠随机分为假手术组(Sham组)和模型组(UUO组),每组16只.假手术组,仅游离输尿管不结扎,模型组,行单侧输尿管结扎术,分别于术后3、7、14 d处死大鼠并留取肾脏组织.肾脏标本行HE、Masson染色方法检测间质纤维化损伤的程度.应用免疫组织化学方法检测肾组织中EMMPRIN,α-SMA,E-cadherin的表达.结果 HE染色显示:UUO术后肾间质肾小管萎缩、扩张,胞外基质沉积,炎细胞浸润增加.Masson染色显示:UUO术后大鼠肾组织胶原纤维组织增生,出现明显的间质纤维化改变.免疫组织化学结果显示:与假手术组相比,UUO模型3d组的EMMPRIN表达显著增高,同时有α-SMA表达升高和E-cadherin表达显著下降(P＜0.05);随着梗阻时间的延长,模型7d组EMMPRIN的表达逐渐减弱(P＜0.05).结论 EMMPRIN可能参与了UUO大鼠肾间质纤维化的早期上皮间质转分化过程,而随着纤维化程度加重,后期表达逐渐减少.     

单侧输尿管梗阻的大鼠内质网应激相关蛋白表达上调

目的探究内质网应激(ERS)相关凋亡通路在单侧输尿管梗阻(UUO)大鼠肾间质纤维化中的作用机制。方法将大鼠随机分为假手术组及UUO模型组,UUO组行左侧输尿管结扎术,于术后3、7和14 d HE和Masson染色观察肾脏病理变化;眼底静脉丛取血,分离血清测定血尿素氮及肌酐;Western blot检测葡萄糖调节蛋白78(GRP78)、内质网源性转录因子(CHOP)、凋亡相关蛋白半胱氨酸天门冬氨酸蛋白酶3(caspase-3)及caspase-12蛋白表达。结果与假手术组相比,UUO模型组可见:1)肾小管扩张和肾间质纤维化程度随输尿管梗阻时间延长而渐进性加重;2)GRP78、CHOP、caspase-3及caspase-12蛋白表达在术后3 d均有上调,随着梗阻时间延长,上述蛋白表达更显著(P0.01)。结论内质网应激相关标志性蛋白在UUO大鼠肾间质纤维化的早期即存在表达上调,可能促进了肾间质纤维化不断进展。     

血清应答因子抑制剂CCG-1423减轻单侧输尿管结扎大鼠肾小管间质纤维化

目的探讨血清反应因子(SRF)抑制剂CCG-1423对单侧输尿管结扎(UUO)大鼠肾间质纤维化的影响。方法将72只无特定病原体(SPF)级SD大鼠中54只行单侧输尿管结扎术(UUO组),另外18只为假手术组(sham组)。UUO组54只大鼠又均分为不注射抑制剂组(空白对照组)、注射0.015 mg/(kg·d)CCG-1423低剂量组[腹腔注射CCG-1423,0.015 mg/(kg·d)]、0.03 mg/(kg·d)CCG-1423高剂量组[腹腔注射CCG-1423,0.03 mg/(kg·d)]。每组分别于术后第3、7、14天,各处死6只大鼠并留取肾脏组织,应用Masson染色检测肾脏纤维化程度,免疫组织化学染色检测肾小管上皮细胞中血清应答因子(SRF)、磷酸化的血清应答因子(p-SRF)、α平滑肌肌动蛋白(α-SMA)、转录因子Snail、上皮性钙黏素(E-cadherin)的表达变化。结果在UUO大鼠模型中随着时间延长纤维化程度逐渐加重,SRF、p-SRF、α-SMA、Snail的表达逐渐增强,E-cadherin的表达逐渐下降。在腹腔注射CCG-1423后Masson染色显示肾脏纤维化减轻,SRF、p-SRF、α-SMA、Snail表达下降,E-cadherin表达增强。结论CCG-1423可抑制UUO大鼠肾组织中SRF的表达,并可减轻肾小管间质纤维化。     

p27和TGF-β在大鼠肾间质纤维化中的表达及关系

目的:研究单侧输尿管梗阻(UUO)大鼠肾组织p27的表达, 同时观察TGF-βmRNA在各组的变化, 探讨UUO模型中 TGF-β与p27的关系.方法:60只SD大鼠随机分成假手术组(SOR)、 UUO模型组.于术后第7、 14、 21天分别处死各组大鼠10只.用HE染色动态观察肾脏病理变化, 免疫组织化学法测定p27的蛋白表达及动态变化, RT-PCR法测定p27mRNA和TGF-βmRNA的水平.结果:SOR组肾小管上皮细胞p27蛋白及肾皮质p27mRNA的表达均强, UUO组随着间质纤维化程度的加重, p27的表达逐渐减弱, 而TGF-βmRNA随着肾间质纤维化程度的加重, 其表达呈上升趋势;UUO组TGF-βmRNA与p27mRNA成负相关.结论:p27参与了UUO大鼠肾间质纤维化的发病过程;UUO大鼠肾间质纤维化模型中TGF-β促纤维化机制可能与p27的下调相关.     

单侧输尿管梗阻大鼠模型中p38MAPK表达与细胞凋亡

目的： 探讨p38MAPK在大鼠单侧输尿管梗阻(UUO)模型肾组织中的表达及其在肾小管间质细胞凋亡和纤维化过程中可能发挥的作用。方法: 将25只Wistar大鼠中的18只行左侧输尿管结扎术，另外7只行假手术。分别于术后第3、7和14 d处死各组大鼠，行HE和Masson染色，观察肾脏病理变化；免疫组织化学方法测定增殖细胞核抗原（PCNA）表达；原位末端标记法（TUNEL）与DNA电泳观察肾小管间质细胞凋亡情况；Western blotting检测肾组织半胱氨酸天门冬氨酸蛋白酶-3（caspase-3）和p38丝裂素激活蛋白激酶（p38MAPK）及其磷酸化产物（p-p38MAPK）的表达。结果: 与假手术组比较，UUO模型组肾脏病理改变加重，TUNEL染色及DNA琼脂糖凝胶电泳可见大量的肾小管间质细胞凋亡，肾间质细胞PCNA表达明显增加，肾组织caspase-3的表达也显著增加（P<0.05）。p38MAPK蛋白水平在各组之间比较没有明显差别（P>0.05）。p-p38MAPK蛋白在正常肾脏有低水平表达，UUO模型组随着梗阻时间延长表达逐渐增多；第7 d达高峰，第14 d开始下降（P<0.05）。结论: p38MAPK信号通路可能参与了UUO致肾小管间质细胞凋亡和肾间质纤维化过程。     

单侧输尿管梗阻大鼠肾脏岩藻糖基转移酶8的表达变化

目的: 探讨岩藻糖基转移酶8(FUT8)在单侧输尿管梗阻(UUO)大鼠模型肾脏中表达及意义。方法: 90只雄性Wistar 大鼠随机分为正常对照组(control组)、假手术组(sham组)和UUO组,分别在术后1 d、3 d、7 d、14 d和21 d处死,检测各组大鼠肾功能情况;PAS与Masson染色观察大鼠肾间质病理形态学改变;RT-PCR和Western blotting检测各组大鼠肾组织不同时点FUT8 mRNA及蛋白的表达;免疫荧光检测大鼠肾组织不同时点FUT8和活化素受体样激酶5(ALK5)的分布以及表达;免疫组化检测大鼠肾脏纤维连接蛋白(FN)、Ⅰ型胶原(Col I)和α-平滑肌肌动蛋白(α-SMA)表达。结果: 与control组相比,UUO组大鼠血肌酐及尿素氮于术后3 d开始升高(P<0.05),第21 d升至最高(P<0.01);UUO3 d开始肾间质可见明显炎症细胞浸润,14 d出现明显肾小管萎缩,21 d可见明显肾间质纤维化。FUT8的mRNA表达与蛋白水平表达于术后3 d出现上升并随梗阻时间延长逐渐增加(P<0.05),分别于14 d及21 d达到最高峰(P<0.01); ALK5在肾小管的表达与FUT8一致,呈正相关(r=0.87,P<0.05),且两者分布一致,共表达区域随着肾纤维化进展逐渐增加。FN、Col I和α-SMA表达于术后第3 d出现上升并随梗阻时间延长逐渐增加(P<0.05),且与FUT8呈正相关(r=0.80,r=0.76,r=0.89,P<0.05)。结论: 在肾间质纤维化的进展中,FUT8与ALK5表达呈时空一致性升高,可能与肾间质纤维化程度密切相关。     

单侧输尿管结扎对大鼠肾间质PI3K/AKT表达的影响

目的探讨PI3K/AKT信号通路是否参与肾间质纤维化的发病过程。方法将30只SD大鼠随机分为假手术组、UUO模型组,模型组行左侧输尿管结扎术。术后5、10和15 d分别处死各组中的5只大鼠,取左肾行HE和Masson染色,并用免疫组化方法检测肾小管-间质中TGF-β1、AKT1、磷酸化AKT1的表达。结果模型组大鼠肾小管-间质细胞增多,肾小管基底膜增厚皱缩,纤维化明显,随时间进展而加重。不同时间的UUO模型组肾组织AKT1、磷酸化AKT1、TGF-β1表达显著高于假手术组(P<0.01);于第10天模型组AKT1表达达高峰,于第15天磷酸化AKT1、TGF-β1表达达高峰;UUO大鼠肾组织第5天和第15天TGF-β1的表达与AKT1活性呈正相关,第10天和第15天梗阻肾的AKT1活性与肾小管-间质纤维化指数呈正相关(均P<0.05)。结论AKT1、磷酸化AKT1在大鼠梗阻性肾病模型肾小管-间质中表达及AKT1活性明显增加,提示PI3K/AKT信号通路可能参与肾小管-间质纤维化的过程。     

PAX2-siRNA在梗阻性肾病大鼠体内的作用研究

目的 通过RNA干扰和基因转染技术,研究沉默PAX2基因对UUO大鼠肾间质纤维化的影响.方法 将PAX2-siRNA转染至UUO大鼠肾脏被膜下,将64只幼年雄性大鼠随机分为:阴性对照组(NC)32只;沉默组(RNAi) 32只,于转染后3、5、7、14天分为4组,每组8只,留取肾组织标本,应用Real-Time PCR及Western blot检测肾皮质PAX2 mRNA及其蛋白的沉默情况,HE染色和Masson染色方法在光镜下观察肾小管损伤情况和肾间质纤维化程度.结果 ①沉默组与对照组比较PAX2 mRNA和PAX2蛋白表达量降低,有统计学意义(P＜0.05),②HE和Masson染色观察到对照组随梗阻时间延长,肾小管损伤逐渐明显,肾间质中可见胶原蛋白逐渐增加.沉默组与对照组比较在7d以前无明显变化(P＞0.05);在梗阻14d沉默组与对照组比较肾小管损伤减轻、肾间质病变减低、胶原纤维沉积减少(P＜0.05).结论 在UUO大鼠模型中,梗阻早期沉默PAX2基因对肾间质纤维化进程无明显影响,梗阻晚期可减轻肾小管损伤及肾间质纤维化病变,对肾间质纤维化有明显治疗作用.     

NO对梗阻性肾脏疾病非血流动力学保护作用机制的研究

目的探讨一氧化氮(nitric oxide,NO)对单侧输尿管梗阻(unilateral ureteral obstruction,UUO)大鼠肾脏间质纤维化的作用及其可能的非血流动力学作用机制。方法成熟Sprague-Dawley雄性大鼠24只,随机分为4组:Ⅰ组假手术组、Ⅱ组模型组、Ⅲ组L-精氨酸(L-Arginine)组、Ⅳ组L-NG-硝基精氨酸甲酯(L-NAME)组。UUO术后28d处死各组大鼠,应用病理染色技术检测肾组织小管间质损伤指数,免疫组织化学法检测肾组织纤维连接蛋白(fibronectin,FN),RT-PCR技术检测肾组织精氨酸酶(Ar-ginase)mRNA的表达。结果与Ⅰ组相比,Ⅱ组肾脏小管间质损伤指数及肾脏组织FN的表达增加(P<0.01);与Ⅱ组相比,Ⅲ组损伤指数及FN的表达减少(P<0.01),Ⅳ组增加(P<0.01)。与Ⅰ组相比,Ⅱ组肾组织Arginase mRNA表达升高(P<0.01);与Ⅱ组相比,Ⅲ组Arginase mRNA表达降低(P<0.05),Ⅳ组升高(P<0.01)。结论NO通过减少细胞外基质(extracellular matrix,ECM)增生延缓肾脏间质纤维化,其对肾脏的保护机制可能部分是通过降低肾脏组织中Arignase mRNA的表达而实现的。     

促红细胞生成素对大鼠肾间质纤维化发动蛋白-1表达影响

目的探讨促红细胞生成素（EPO）对大鼠肾间质纤维化发动蛋白．1（Drp-1）表达的影响。方法81只sD大鼠,分为假手术组、对照组和EPO组,每组各27只。EPO组、对照组采用单侧输尿管结扎并剪断建立肾间质纤维化模型,假手术组仅游离输尿管而不结扎和剪断;EPO组予EPO皮下注射;假手术组和对照组给予等量生理盐水皮下注射。各组于术后7、14和21d取动脉血分离血清检测血肌酐和尿素氮水平。取梗阻侧肾组织行苏木精．伊红和Masson染色,观察肾脏病理学变化。用免疫组化方法检测肾组织Drp一1的表达。结果@EPO组各时点血清肌酐和尿素氮水平显著低于对照组（P〈0．01）,但高于假手术组（P〈0．05）。②EPO组各时点肾组织病理改变较对照组明显减轻,肾小管间质损伤评分和肾间质纤维化相对面积均低于对照组（P均〈0．05）,但高于假手术组（P〈0．05）。③EPO组肾组织的Drp．1表达显著低于对照组（P〈0．05）,但高于假手术组（P〈0．05）。结论Drp-1在大鼠肾间质纤维化肾组织中表达增加,参与。肾脏纤维化过程;EPO可以通过抑制Drp一1表达,从而延缓肾间质纤维化的进展。     

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.     

Intestinal helminthiasis in school children in Haiti in 2002

A survey on intestinal helminths in school children was conducted in Haiti in 2002. This first nationwide study involving the entire country was stratified by department according to urban and rural zones using the cluster method. Focusing on elementary school children (n=5792; age range 3 to 20 years), it involved 26 urban and 49 rural schools randomly selected. Stools were preserved in formalin and examined by the Ritchie technique. Thirty-four per cent of stools (1981/5792) tested positive for intestinal helminths with the following parasites identified: Ascaris lumbricoides (27.3%), Trichuris trichiura (7.3%), Necator americanus (3.8%), Hymenolepsis nana (2%), Taenia sp. (0.3%) and Strongyloides stercoralis (0.2%). The helminth prevalence was higher in rural (38.4%) compared to urban areas (30%). There was no significant difference in prevalence by sex and age. The importance of geohelminths changed from one department to another with the highest prevalence found in the Southern department of Grande Anse (73.7%) and the lowest prevalence in the Center department (20.6%). Five out of the country's nine departments had a similar prevalence varying from 25.5% to 28.2%. Intestinal helminthic polyparasitism was observed in a percentage of infested school children comprise between 3.4% and 28.6% according in relation to the geographical area. A program to fight against geohelminths in school children should be initiated as a public health priority. Albendazole is the drug of choice. Frequency of drug distribution should be based on the prevalence of geohelminths in each department.     

Experience in adult population in dengue outbreak in Delhi

A dengue outbreak has recently hit the Indian capital. We studied the clinical profile of adult patients. Five hundred and sixty patients of dengue infection were admitted in a specially created ward according to the criteria laid down by WHO. Haematemesis (28.28%), epistaxis (26.78%) and malena (14.28%) were some of the common presentations. Similarly lymphadenopathy, especially cervical (30.89%), palatal rashes (26.96%) and hepatomegaly (23.75%) were the most commonly encountered findings on physical examination. Most of the cases were of dengue fever with haemorrhage and only 2.5% cases were classified under dengue haemorrhagic fever or dengue shock syndrome. The average hospital stay was 3.4 days but only 9.8 hours in the eleven patients who died, suggesting their late arrival in preterminal situation giving little time for resuscitation. Thrombocytopenia was not a feature and only 12.85% patients had platelet count less than 70,000/cmm. Most of the patients who were admitted with thrombocytopenia, showed normalization in their platelet counts in next few days. Serological examination demonstrated evidence of recent dengue infection in 41.17% patients. Few patients required blood or platelet concentrate transfusion. Eleven patients died, three due to DIC, one of intracranial haemorrhage and seven due to massive gastric haemorrhage. Rest of the patients recovered completely. Thus we can conclude that recent outbreak in Delhi was of dengue fever with haemorrhage and mortality was very low in patients who came early to the hospital.     

Changes in the hypothalamic nuclei in in experimental allergy

Summary In rabbits subjected to prolonged sensitization and in which the Arthus phenomenon was induced there was a marked reaction of the hypothalamic nuclei. Staining by Gomori's method indicated a cellular swelling, loss of granules, and protoplasmic vacuolization in the supraoptic nucleus. There was a considerable increase in the size of the cross-sectional area of the cells. The same effects were much less well shown in the paraventricular nucleus. These results show that marked signs of increased neurosecretion developed in the animals at the height of the Arthus phenomenon.(Presented by Active Member AMN SSSR V. V. Parin) Translated from Byulleten Éksperimental'noi biologii i Meditsiny, Vol. 55, No. 4, pp. 110–113, April, 1963.     

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.