高脂饮食上调兔子血管内膜与血清炎性因子表达

目的:分析高脂饮食致肥胖或动脉粥样硬化兔子血管内膜白细胞介素-1β(IL-1β)和肿瘤坏死因子-α(TNF-α)水平变化。方法:12只健康新西兰大白兔,分为普通饮食组(OD组)和高脂饮食组(HFD组)。依据动脉粥样硬化造模方法,高脂饮食喂养兔8周后,获取兔子血液或颈动脉组织。检测血细胞相关参数和血液生化参数及动脉内膜的厚度,同时用ELISA检测IL-1β和TNF-α水平,Western Blot法检测血管内膜IL-1β和TNF-α蛋白的表达。比较分析两组各检测参数的差异。结果:两组兔子外周血细胞检测项目无统计学差异(P>0.05)。HFD组脂质检测指标甘油三酯(TG)、总胆固醇(TC)、低密度脂蛋白胆固醇(LDL-C)和载脂蛋白B(ApoB)均显著高于OD组(P<0.05,P<0.01),而高密度脂蛋白胆固醇(HDL-C)则显著低于OD组(P<0.05)。其它血液生化项目在两组间的水平无显著差异(P>0.05)。ELISA检测OD组血浆IL-1β和TNF-α的水平显著低于HFD组(P<0.05)。以高脂饮食喂养健康兔子发现HFD组兔血管内膜的厚度明显高于OD组(P<0.05),且血管内膜炎性因子IL-1β和TNF-α蛋白表达具有同样的趋势。结论:以高脂饮食喂养健康兔子发现动脉粥样硬化模型兔子炎性因子IL-1β和TNF-α在血液中和血管内膜均有表达增高,或可与动脉粥样硬化相关。     

氯吡格雷对兔髂腹动脉球囊损伤后新生内膜的影响

目的： 观察氯吡格雷对兔髂腹动脉球囊损伤后内膜增生的影响和作用机制。方法: 37只新西兰兔随机分为正常组、模型组、氯吡格雷组及阿托伐他汀组。高脂喂养加髂动脉内膜剥脱建立动脉粥样硬化模型。检测血脂和高敏C反应蛋白（hsCRP）。12周后观察动脉病理组织学改变并测量内膜、中膜厚度和面积，原位杂交检测血小板源性生长因子（PDGFmRNA）的表达，免疫组化测定凋亡基因Bcl-2和Bax的表达。结果： ①模型组血清hs-CRP、髂动脉内膜厚度和面积、PDGFmRNA、凋亡基因Bcl-2/Bax比值较正常组显著增加（P＜0.05,P＜0.01）；②氯吡格雷组血清hs-CRP、内膜厚度与面积、PDGFmRNA的表达、Bcl-2/Bax的比值小于模型组（P＜0.05,P＜0.01），血脂水平和模型组无明显差异；③氯吡格雷对上述指标的改变与阿托伐他汀无显著差异。结论： 氯吡格雷降低炎症反应、抑制PDGFmRNA表达、促进细胞凋亡，减轻高脂和球囊损伤后动脉内膜增殖。     

T淋巴细胞对高脂饮食引起的肠道炎症的影响

目的:探讨T淋巴细胞在高脂饮食导致的肠道炎症中所起的作用。方法:采用了T淋巴细胞缺失(CD3ε敲除)的小鼠模型,C57BL/6J野生型小鼠作对照,高脂喂养16周。通过实时荧光定量PCR(qRT-PCR)检测结肠组织中的炎性因子水平,苏木精-伊红(HE)染色直接观察肠道形态,检测肠道肌层厚度,同时通过免疫组织化学技术(IHC)观察肠道组织中免疫细胞的浸润情况。结果:高脂饮食饲喂小鼠16周后,小鼠体重明显增加。qRT-PCR检测显示,与正常饮食的野生型小鼠组相比,高脂饮食的野生型小鼠结肠组织中IL-1β(P0. 01)、IL-5(P0. 05)、TNF-α(P0. 05)、IFN-γ(P0. 05)以及IL-6(P0. 05)表达水平明显上调; HE染色显示高脂饮食组结肠肌层明显变薄,同时免疫组化的结果显示结肠组织中巨噬细胞浸润增强。以高脂饮食野生型小鼠为对照,高脂饮食的T淋巴细胞缺失小鼠的结肠组织中IL-1β(P0. 01)、IL-6(P0. 05)、IFN-γ(P0. 05)、IL-5(P0. 05)以及TNF-α(P0. 01)基因水平下调; HE染色显示高脂饮食的T淋巴细胞缺失小鼠结肠肌层增厚,同时免疫组化结果显示结肠组织中巨噬细胞浸润减少。结论:T淋巴细胞对高脂饮食导致的肠道炎症有促进作用。     

高脂饲料喂养加静脉注射小牛血清白蛋白建立兔动脉粥样硬化模型**

背景：目前单纯饲喂高脂饲料建立动脉粥样硬化狭窄模型较常见。 目的：采用高脂饲料喂养加静脉注射小牛血清白蛋白建立兔动脉粥样硬化模型。 方法：分别单纯高脂饮食、高脂饲料+脂肪乳灌胃以及高脂饲料+小牛血清白蛋白3种不同的方法喂养兔构建动脉硬化模型,设立正常对照组,予普通饲料喂养。 结果与结论：各组兔血清胆固醇、三酰甘油、低密度脂蛋白和高密度脂蛋白水平较正常对照组显著升高(P < 0.01)。单纯高脂组可出现高脂血症,未发现典型的动脉粥样硬化病变;高脂+脂肪乳灌胃组可形成纤维斑块;高脂+小牛血清白蛋白组可形成较成熟的动脉粥样硬化斑块。结果证实,高脂饲料加静脉注射小牛血清白蛋白可形成较成熟的动脉粥样硬化斑块,可成功建立动脉粥样硬化兔模型。关键词：动脉粥样硬化;小牛血清白蛋白;高脂饲料;兔;动物模型;组织工程 doi:10.3969/j.issn.1673-8225.2012.20.018     

中药赤芍对球囊损伤术后血管重构的干预研究

目的 观察中药赤芍防止球囊损伤术后血管重构作用。方法 新西兰白兔随机分为对照组、单纯高脂组、赤芍高剂量组、低剂量组。高脂喂养6周建立动脉粥样硬化模型,行颈动脉球囊损伤术,8周时取材作病理形态学检查。结果(1)与高脂组比较,赤芍高、低剂量组增生内膜面积、中层面积,内膜、中膜、外膜PCNA阳性着色均显著减少(P<0.05或P<0.01)。(2)各组内皮增生成分主要为平滑肌细胞;巨噬细胞阳性着色主要分布在外膜,与高脂组比较,赤芍高、低剂量组阳性着色较少。(3)高脂组动脉损伤侧外膜Ⅰ型胶原增多,赤芍组Ⅰ型胶原增加较少。结论 赤芍对高脂喂养兔颈动脉球囊损伤术后血管重构有显著防止作用。     

诱发糖尿病饲料致新西兰兔动脉粥样硬化的作用

目的 :糖尿病是动脉粥样硬化的主要危险因素之一。建立糖尿病促发动脉粥样硬化的动物模型有助于阐明糖尿病促进动脉粥样硬化发生的机制。目前 ,这方面的模型非常稀少 ,为此 ,我们用高脂高糖饲料喂养新西兰兔 ,拟建立一种新的糖尿病并发动脉粥样硬化动脉模型。方法 :将新西兰兔分为二组 :正常饲料组 (ｎ =7) ;高脂高糖饲料喂养组 (ｎ =1 0 ,喂含 1 0 %猪油、36%白蔗糖混合饲料 )。共观察 4个月 ,每月取动物禁食过夜空腹血测血糖、胰岛素、总胆固醇、ＨＤＬ -胆固醇、甘油三酯等。结果 :本研究发现 ,实验组一月后即出现高甘油三酯、高血糖、实…     

载脂蛋白E基因敲除小鼠动脉粥样硬化早期血管外膜成纤维细胞增殖活性增强

目的:探讨动脉外膜成纤维细胞增殖与早期动脉粥样硬化病灶形成的关系。方法:选择6周龄载脂蛋白E基因敲除[apoE(-/-)]小鼠和野生型C57BL/6小鼠,高脂喂养2、4和10周后,在各个时点处死动物前24 h经腹腔注射5-溴-2-脱氧尿嘧啶(BrdU),后选取升主动脉制备连续切片,通过HE染色观察组织形态学的变化,用免疫组化方法观察不同时点血管外膜及内膜BrdU的表达变化。体外培养高脂喂养2周的apoE(-/-)小鼠和C57BL/6小鼠动脉外膜成纤维细胞,通过BrdU掺入法测定细胞增殖活性,流式细胞术测定细胞周期。结果:体内实验发现apoE(-/-)小鼠高脂喂养2周后,在无可见内膜病灶形成之前,首先在主动脉外膜发现BrdU标记的阳性细胞,之后才在损伤内膜观察到BrdU标记细胞。而C57BL/6小鼠在任何时点都未检测到BrdU标记的细胞。体外实验观察到apoE(-/-)小鼠血管外膜成纤维细胞BrdU标记的细胞数显著多于C57BL/6小鼠(P0.01),apoE(-/-)小鼠血管外膜成纤维细胞S期及G2/M期所占百分比明显高于对照组(P0.05)。结论:血管外膜成纤维细胞增殖可能参与早期动脉粥样硬化病灶形成。     

细胞外基质金属蛋白酶诱导因子与尿激酶型纤溶酶原激活物在载脂蛋白E 敲除大鼠动脉粥样硬化斑块表达的意义及相关性*

目的：探究细胞外基质金属蛋白酶诱导因子（ EMMPRIN）与尿激酶型纤溶酶原激活物（ μPA）在载脂蛋 白E 基因（ ApoE）敲除大鼠动脉粥样硬化斑块表达意义及相关性。方法：选择ApoE 敲除大鼠,将其分为对照组 与高脂饮食组,对照组大鼠实施正常饮食喂养,高脂饮食组实施高脂饮食喂养,并分别于喂养的第6、10、14、 18 周处死部分大鼠,采用H-E 染色观测动脉粥样硬化斑块形态, 免疫印迹及RT-PCR 检测主动脉粥样硬化斑块内 EMMPRIN和μPA 的表达及其mRNA表达,并实施组间比较。结果：高脂饮食组大鼠动脉出现明显动脉粥样硬化 斑块,且随着时间的推进,逐渐加重,而对照组大鼠并未出现明显的动脉粥样硬化斑块;喂养第6、10、14、18 周时高脂饮食组大鼠主动脉内EMMPRIN和μPA 的表达及其mRNA表达均明显高于对照组;EMMPRIN与μPA 表 达呈正相关。结论： EMMPRIN与μPA 在动脉粥样斑块中的表达水平呈正相关关系,两者可能参与动脉粥样硬化 斑块的形成并发挥促进作用。     

免动脉粥样硬化病理定量分析对血管内超声显像结果的验证

目的 :应用光镜和电镜病理检查技术验证血管内超声显像 (introvascularultrasound ,IVUS)对动脉粥样硬化的诊断。方法 :以IVUS技术探测粥样斑块的位置、性质和大小 ,取材后进行HE、Hart法染色 ,用测微尺测量血管壁及斑块厚度 ,然后进行量化分析 ,辅以电镜技术观察病变的形态学改变。结果 :经组织形态学观察 ,对照IVUS探测结果 ,弹性动脉是否存在斑块的符合率达 94%。对照组 12个血管位点均未见斑块 ,实验组 33个位点病理发现 32个有斑块 ,IVUS发现 30个有斑块。对斑块厚度的测量二者相关性较好 (r =0 835 ) ,差异有显著性 (P <0 0 5 ) ;对血管中膜厚度的测量二者亦有相关性 (r =0 814) ,差异有显著性 (P <0 0 5 ) ;对正常血管内膜测量二者相关性差 (r =0 0 5 5 ) ,不具有差异显著性 (P >0 0 5 )。结论 :从组织形态学的验证和量化分析可知IVUS能够准确判断弹性动脉粥样硬化及动脉中膜厚度 ,但对于动脉内膜厚度的测量 ,IVUS不如病理测量准确。     

紫薯花色苷提取物对ApoE~(-/-)小鼠动脉粥样硬化和高脂血症的作用

目的:研究紫薯花色苷提取物对Apo E-/-小鼠动脉粥样硬化和高脂血症的作用。方法:将8周龄雄性ApoE~(-/-)小鼠随机分为对照组、高脂饮食组、花色苷低剂量组、花色苷高剂量组和立普妥组,共5组(n=10),除对照组小鼠饲喂普通饲料外,其它4组在饲喂高脂饲料8周后,再给予蒸馏水、紫薯花色苷提取物(1. 67 mg·kg~(-1)·d~(-1)和8. 35 mg·kg~(-1)·d~(-1))或立普妥(阿托伐他汀钙,2 mg·kg~(-1)·d~(-1))灌胃8周。心脏取血检测小鼠血清的总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)及高密度脂蛋白胆固醇(HDL-C)水平;油红O和HE染色评估主动脉根部斑块面积和肝脏的脂肪变性程度; RT-q PCR方法检测主动脉和肝脏中炎症因子的表达。结果:与对照组比较,高脂饮食组小鼠血清的TC、LDL-C和HDL-C水平明显升高(P 0. 01),动脉的斑块面积明显增大(P 0. 01),肝脏的脂肪变性明显,白细胞介素1β(IL~(-1)β)、IL-6和单核细胞趋化蛋白1(MCP~(-1))的表达增加(P 0. 01),而过氧化物酶体增殖物激活受体α(PPAR-α)的表达降低(P 0. 05);高剂量紫薯花色苷提取物可以明显降低TC、TG和LDL-C水平(P 0. 01),减少动脉的斑块面积(P 0. 01),减轻肝脏的脂肪变性程度,同时还可以降低IL~(-1)β、IL-6和MCP~(-1)的表达(P 0. 05),提高PPAR-α的表达(P 0. 05)。结论:紫薯花色苷提取物可调节血脂,降低炎症细胞因子表达,从而抑制动脉粥样硬化的发展和高脂血症引起的肝组织脂肪变性。     

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.     

Pitfalls in TRAP assay in routine detection of malignancy in effusions

Telomerase has been found to be reactivated in a majority of cancers but is inactive in most somatic cells. Our principal goal was to determine the potential use of the telomeric repeat amplification protocol (TRAP) assay as marker for malignancy in cytological effusions. The simple selection criterion was the cytological diagnosis, and routine samples were classified into malignant (58 samples) and nonmalignant (233 samples). Of the malignant samples, 44/58 (76%) were positive by TRAP assay. Of the 14 telomerase-negative cytology-positive samples, RNA integrity was poor in 9, indicating suboptimal sample conservation for molecular analysis. In 3 of the remaining 5 samples with a negative TRAP assay, a high number of malignant cells was observed, and these cells might have been telomerase-negative. Thus, the sensitivity of TRAP assay for the presence of malignant cells was about 76%. In the cytologically nonmalignant effusions, the presence of telomerase activity was observed in 24% (55/233). Of these, 6% were highly suspicious for malignancy, 9% were doubtful, and 9% were cytologically nonmalignant effusions confirmed by a follow-up of 12 mo or more. According to these data, the specificity of the TRAP assay to detect tumor cells in effusions ranged only between 82-91%. Our results indicate that, although the TRAP assay is positive in 6-15% of putative malignant effusions, the relatively high number of TRAP false-negative and false-positive cases renders this test unsuitable for routine diagnostic purposes.