阿托伐他汀对高糖高脂环境下人脐静脉内皮细胞与人肾小球系膜细胞相互作用的影响

 目的 探讨高糖高脂环境下人脐静脉内皮细胞与人肾小球系膜细胞间的相互作用及阿托伐他汀对内皮细胞与系膜细胞相互作用的影响。方法人脐静脉内皮细胞与人肾小球系膜细胞共培养和人肾小球系膜细胞单独培养,分为对照组、甘露醇组、高糖高脂组、BN52021组、阿托伐他汀组。ELISA法检测细胞上清液纤维联接蛋白（Fn）、血小板活化因子（PAF）含量,实时荧光定量检测系膜细胞血小板活化因子受体（PAF-R）mRNA表达。结果 (1)高糖高脂条件下,共培养和单培养Fn、PAF升高（P <0.05）;高糖高脂条件下,共培养Fn、PAF较单培养升高（P <0.05）,BN52021和阿托伐他汀可抑制高糖高脂引起的Fn升高（P <0.05）,阿托伐他汀可抑制高糖高脂引起的PAF升高（P <0.05）;（2）高糖高脂可上调系膜细胞PAF-R mRNA表达（P <0.05）,阿托伐他汀可显著抑制PAF-R mRNA表达上调（P<0.05）。结论 (1) 高糖高脂环境下,系膜细胞和内皮细胞存在异常的相互作用。(2)阿托伐他汀可影响高糖高脂环境下内皮细胞与系膜细胞的相互作用。     

细胞相互作用对PAF、系膜细胞paf-R基因表达的影响及阿托伐他汀的干预作用

目的:探讨高糖高脂条件下内皮细胞与系膜细胞相互作用对PAF产生、系膜细胞paf-R基因表达的影响及阿托伐他汀的干预作用。方法:内皮细胞与系膜细胞共培养和系膜细胞单独培养后随机,分为对照组、甘露醇组、高糖高脂组、阿托伐他汀组,培养24h后,ELISA法检测各组细胞上清液PAF含量,实时荧光定量检测系膜细胞paf-RmRNA表达。结果:(1)高糖高脂促进共培养组和单培养组PAF升高(P0.05),共培养组PAF均较单培养组升高(P0.05);(2)高糖高脂可上调系膜细胞paf-R mRNA表达(P0.05);(3)共培养和单培养条件下,阿托伐他汀可抑制高糖高脂引起的PAF升高(P0.05),并可抑制高糖高脂引起的系膜细胞paf-RmRNA表达上调(P0.05)。结论:(1)高糖高脂环境下,系膜细胞和内皮细胞存在异常的相互作用,这种作用促进PAF产生;(2)高糖高脂促进系膜细胞paf-R基因表达,使PAF的生物效应进一步放大;(3)阿托伐他汀可影响高糖高脂条件下内皮细胞与系膜细胞之间的相互作用。     

血小板活化因子受体拮抗剂对实验性肾病综合征的疗效观察及机制探讨

目的:探讨血小板活化因子受体拮抗剂对实验性肾病综合征的疗效及机制。方法: 实验大鼠均复制成阿霉素(ADR)肾病模型,分为2组:ADR肾病组和ADR肾病+血小板活化因子(PAF)受体拮抗剂(BN52021)组。结果: ADR肾病+BN52021组大鼠各期尿蛋白量、血清总蛋白下降幅度、血清胆固醇上升幅度均显著低于ADR肾病组(P＜0.05),第21 d时血清肌酐含量显著低于ADR肾病组(P＜0.05);电镜下肾组织病理改变显著轻于ADR肾病组。ADR肾病组中,肾皮质PAF最大产量在14 d(先于最大蛋白尿量),而肿瘤坏死因子(TNFα)最大含量在21 d,而且, ADR肾病+BN52021组肾皮质内PAF、TNFα含量显著低于ADR肾病组。结论: PAF可能直接或间接通过刺激肾小球固有细胞(肾小球系膜细胞、上皮细胞等)产生TNF导致肾小球损伤,PAF拮抗剂可能通过抑制肾皮质内PAF、TNFα合成,而减轻肾小球损伤。     

血小板活化因子受体在SAP胰腺组织的变化及银杏苦内酯B的影响

目的探讨血小板活化因子受体(PAF-R)在重症急性胰腺炎(SAP)胰腺组织的动态变化及银杏苦内酯B(BN52021)干预前后的影响。方法Wistar大鼠随机分为阴性对照组(NC组)、SAP模型组(SAP组)、BN52021治疗组(BN组),每组按手术后不同时相点(1、2、3、6、12和24h)分为6小组,分别测定血淀粉酶的变化,并对胰腺组织进行病理学观察和评分,用Westernblot检测胰腺组织PAF-R蛋白质表达的变化。结果BN52021能降低SAP的血淀粉酶和改善其病理变化;SAP组和BN组胰腺组织PAF-R在3h达到高峰(P<0.05),除1h外其余各时相点与NC组有显著性差异(P<0.05);BN组与SAP组之间无显著性差异。结论PAF-R在SAP过程中显著升高,对SAP的病情发展有重要意义,BN52021对PAF-R蛋白表达无显著影响,可能通过竞争性抑制上升的PAF与表达量同时上升的PAF-R相结合,实现它的治疗作用。     

高糖诱导大鼠系膜细胞CTGFmRNA的表达及机制探讨

目的： 观察高糖状态对肾小球系膜细胞中p38丝裂原活化蛋白激酶（p38 MAPK）及其下游因子cAMP反应元件结合蛋白1（CREB1）的表达以及对系膜细胞结缔组织生长因子（CTGF）和细胞外基质蛋白表达的影响。方法: 体外培养大鼠肾小球系膜细胞,分别给予高糖和p38 MAPK抑制剂SB203580干预,采用Western印迹检测p38 MAPK和CREB1及其磷酸化蛋白(p-p38 MAPK、p-CREB1)的表达,半定量RT-PCR检测CTGF和纤维黏连蛋白（FN） mRNA的表达。放免法测定细胞上清液中层黏连接蛋白(LN)和IV型胶原的分泌含量。结果: 与低糖组和低糖加甘露醇组相比,高糖组系膜细胞p-p38 MAPK、p-CREB1表达明显上调,CTGF和FN mRNA的表达增加,细胞上清液中FN和IV型胶原含量增加。SB203580能够明显抑制高糖培养系膜细胞p38 MAPK和CREB1的磷酸化,下调CTGF和FN mRNA表达,抑制细胞上清LN和Ⅳ型胶原的分泌。结论: p38 MAPK信号途径可能参与高糖诱导的肾小球系膜细胞CTGF的表达和细胞外基质的分泌。     

Caveolae在高糖诱导的大鼠肾小球系膜细胞细胞外基质合成中的作用

 目的：探讨胞膜小凹(caveolae)在高糖(HG)诱导大鼠肾小球系膜细胞(MCs)细胞外基质(extracellular matrix,ECM)表达过程中的作用。方法：传代培养的大鼠MCs同步化后分为：(1)正常糖组;(2)HG组;(3)HG+甲基-β-环糊精(β-MCD)组;(4)HG+β-MCD+胆固醇组。用Western blotting检测小凹蛋白1(Cav-1)、磷酸化小凹蛋白1(p-Cav-1-Y14)和Ⅰ型胶原(ColⅠ)蛋白的表达水平,用实时定量PCR检测细胞中Cav-1 mRNA表达变化,用ELISA方法检测上清中纤维连接蛋白(FN)的蛋白含量。结果： (1)高糖状态下, 系膜细胞 FN和ColⅠ 蛋白表达水平均显著增加(均P<0.05)。(2)高糖培养显著增加p-Cav-1-Y14水平(P<0.01),而对Cav-1 mRNA及蛋白表达无明显影响(P>0.05)。(3)β-MCD预处理抑制了高糖诱导的 p-Cav-1-Y14升高(P<0.01)及FN表达(P<0.05),但对高糖诱导的Col I表达无影响;β-MCD的作用可被胆固醇抑制。结论：高糖可显著增加系膜细胞1型胶原及纤维连接蛋白的合成。高糖诱导的纤维连接蛋白合成与Cav-1磷酸化水平增加有关。     

Decorin 基因转染抑制高糖培养的大鼠肾系膜细胞细胞外基质mRNA表达

目的： 观察核心蛋白聚糖Ⅱ（DCN）基因转染对高糖培养的大鼠肾系膜细胞（RMCs）细胞外基质（ECM）mRNA表达的影响。方法： 用已构建的AD-DCN腺病毒感染高糖环境培养的大鼠肾系膜细胞，采用半定量RT-PCR测定decorin、转化生长因子β₁（TGF-β₁）、纤维粘连蛋白（fibronectin）和IV型胶原（collagen IV） mRNA水平。结果： 高糖培养的RMCs感染AD-DCN后，DCN mRNA水平显著增高，24 h至第7 d，该组DCN mRNA水平始终显著高于高糖组及AD－LacZ感染组(P<0.05)，decorin mRNA水平增高后表现出对TGF-β₁ mRNA 水平的抑制作用，在mRNA水平，AD-DCN感染的RMCs细胞外基质fibronectin和collagen IV随TGF-β₁ mRNA水平下降而下降。结论： DCN重组腺病毒感染大鼠肾系膜细胞后能高效表达目的基因decorin并拮抗 TGF-β₁ 的生物活性，其下调ECM成分 mRNA水平的作用与使用TGF-β抗体的效果相似。     

脂蛋白对肾小球膜细胞产生血小板活化因子的影响

我们利用体外培养的正常人肾小球系膜细胞，观察了纯化的人低密度脂蛋白和高密度脂蛋白对该细胞产生上板活化因子的影响。结果发现：经刺激２０ｈ在一定浓度范围内，低密度脂蛋白能增加增减系膜细胞上清液中血小板活化因子的含量，且在浓度为１００μｇ／ｍｌ时，培养细胞上清液中血小板活化因子的上升显著，在２００μｇ／ｍｌ，达到高峰，而高密度脂蛋白则在高浓度（３５０μｇ／ｍｌ）时有轻度刺激作用，这些结果说明：Ｗ氏密度脂     

缬沙坦抑制高糖培养的大鼠肾小球系膜细胞细胞外基质及CTGF的表达

目的　探讨高糖状态下体外培养的肾小球系膜细胞中结缔组织生长因子（CTGF）的表达以及缬沙坦的影响。方法 体外培养大鼠系膜细胞,分别给予高糖和缬沙坦干预,采用Western blot检测CTGF、p38丝裂原活化蛋白激酶 （p38 MAPK）、cAMP反应元件结合蛋白1（CREB1）及各自磷酸化蛋白的表达,RT-PCR检测CTGF mRNA和纤维黏连蛋白（FN）mRNA的表达。放免法测定细胞上清液中层黏连蛋白(LN)和IV型胶原的含量。结果 与低糖组相比,高糖组系膜细胞CTGF、p-p38 MAPK、p-CREB1表达明显上调,CTGF mRNA和FN mRNA表达增加,细胞上清中LN和IV型胶原含量增加。缬沙坦组CTGF、p-p38 MAPK、p-CREB1的表达明显下调,CTGF mRNA和FN mRNA表达降低, LN和IV型胶原的含量减少。 结论　缬沙坦抑制高糖状态系膜细胞CTGF表达 和细胞外基质的分泌可能部分是通过影响p38 MAPK及其下游核因子CREB1的激活而实现。     

高糖对人肾系膜细胞GSH-PX和PDGF-B表达影响及茶多酚干预作用

目的探讨高糖条件下系膜细胞氧化失衡与血小板源性生长因子—B（PDGF—B）表达之间的关系,通过抗氧化剂茶多酚证实氧化应激在早期糖尿病中的作用。方法将培养的系膜细胞分为正常对照组、高糖组、茶多酚对照组和茶多酚干预组,分别在0、12、24、36、48h采用分光光度法检测上清液还原型谷胱甘肽过氧化物酶（GSH—PX）活性,以及免疫细胞化学法和RT—PCR法检测0、12、36h的PDGF—B mRNA和蛋白的表达情况。结果①高糖条件下GSH—PX活性下降,抗氧化能力下降,氧化失平衡;茶多酚可增加抗氧化酶GSH—PX的活性。②高糖组系膜细胞PDGF—B mRNA和蛋白的表达比正常对照组均增加（P〈0．05）;茶多酚干预组比高糖组有明显下降（P〈0．05）。结论高糖能促进系膜细胞活性氧和PDGF—B的表达增加,茶多酚可抑制该作用。     

Modes of reinforcement and gender and culture of experimenter and subject: effects of "alien" and external reinforcement for Japanese and American men and women

In this study, we examined external and "alien" reinforcement (ER and AR. respectively) as a factor in social learning, and studied the combined effects of culture and reinforcement mode. A female (Experiment 1) and a male (Experiment 2) experimenters conducted experimental sessions. Both men and women, who grew up in the same culture as the experimenter, participated and performed the experimental task. A three-way interaction effect of experimenter gender, culture, and reinforcement mode was found on task performance. And the effect was more pronounced for a Japanese experimenter. A female and a male experimenters conducted Experiments 3 and 4, respectively; however participants this time were men and women who grew up in different cultures than the experimenter. Results indicated that the pattern of the subject gender and reinforcement mode interaction effect, when the experimenter was Japanese with American subjects, was exactly opposite to that when the experimenter was American. These experiments showed that AR was as effective for social learning as ER, and that the cultural backgrounds of experimenter and subject influenced AR and ER effectiveness.     

Rates of oxygen consumption and of anaerobic glycolysis in renal cortex and medulla of adult and new-born rats and guinea-pigs

1. Rates of oxygen uptake and of anaerobic glycolysis were estimated in slices from the renal cortex and medulla (a) of adult rats and guinea-pigs, (b) of new-born (1-, 5- and 21-day-old) rats and of guinea-pigs of 1, 12, 21, 24 and 120 hr age.2. In the adult rat, Q(O2) values for the cortex were 12.55 +/- 0.20 (22) and for the medulla: 8.56 +/- 0.17 (22) mul./hr.mg dry weight, while in the new-born rat (24 hr old) they were 10.99 +/- 0.46 (12) and 9.33 +/- 0.18 (9) mul./hr.mg dry weight respectively.3. Values for Q(CO2) (N2) (anaerobic glycolysis) in the 14 hr old new-born rat were in the renal cortex 9.65 +/- 0.35 (5) and in the medulla 7.39 +/- 0.43 (5) mul./hr.mg dry weight; while in the adult they were 2.25 +/- 0.08 (16) and 5.76 +/- 0.14 (16) mul./hr.mg dry weight, respectively.4. In the adult guinea-pig values for Q(CO2) (N2) were of the same order as in the adult rat, though the rate of O(2) uptake was for the cortex 8.12 +/- 0.22 (12) and for the medulla 5.02 +/- 0.23 (11) mul./hr.mg dry weight.5. Though the Q(O2) values in the renal cortex and medulla were smaller in the 1 hr old new-born guinea-pig, they were already increasing in the 12 hr old neonate.6. The results are discussed in the light of enzyme changes occurring during the process of maturation of the nephron as indicated by histochemical observations.     

Timing and intensity of early fevers and the development of allergies and asthma

BACKGROUND: Early childhood fevers appear to protect against later allergies and asthma. What is not known is the time in which fevers exert this effect and whether the degree of temperature increase is important. OBJECTIVE: We sought to examine the relationship between the time and degree of early fevers and later allergies and asthma. METHODS: Eight hundred thirty-five children from southeast Michigan were enrolled at birth. Clinic records from their first 2 years were abstracted for episodes of fever. At age 6 to 7 years, children underwent allergy testing. We examined fevers occurring within 6-month intervals in the first 2 years of life and outcomes at age 6 to 7 years. The primary outcome measures were allergic sensitization, asthma, asthma with allergic sensitization, and asthma without allergic sensitization. RESULTS: In the unadjusted analysis each episode of fever between 7 and 12 months of age was associated with a lower odds of allergic sensitization (odds ratio [OR], 0.71; 95% CI, 0.54-0.93) and asthma with allergic sensitization (OR, 0.43; 95% CI, 0.21-0.90) at age 6 to 7 years. Likewise, every 1 degrees C increase in the maximum temperature between 7 and 12 months was associated with a lower odds of allergic sensitization (OR, 0.77; 95% CI, 0.61-0.96) and asthma with allergic sensitization (OR, 0.62; 95% CI, 0.40-0.94). After adjusting for potential confounders, each episode of fever between 7 and 12 months was associated with a lower likelihood of asthma with allergic sensitization (adjusted OR, 0.33; 95% CI, 0.11-0.94) at age 6 to 7 years. CONCLUSIONS: Both the timing and intensity of childhood fevers appear to be important factors in the development of allergies and asthma.     

纳米水平分子成像与诊疗一体化研究进展与挑战

分子成像能以非侵入性的方式重现活体细胞的生理功能和生物学过程,提高疾病的早期和特异性诊断水平。纳米颗粒/材料具有物理性质可控性高、易于表面修饰、血液循环时间长和可功能化等优点,在疾病诊断与治疗中显示出巨大潜力。但如何阐明纳米材料多功能间的内在联系、解决其代谢及安全性等关键机制难题、实现纳米颗粒/材料多功能性到临床多功能...