PPARγ对ECV-304细胞ICAM-1与VCAM-1表达的影响

目的 观察过氧化物酶体增殖物激活型受体γ(PPARγ)对糖基化终末产物(AGEs)所诱导的血管内皮细胞表达ICAM-1、VCAM-1的影响,探讨PPARγ在糖尿病血管并发症及动脉粥样硬化中的可能作用.方法 体外培养人内皮细胞系ECV-304,应用PPARγ的反义寡核苷酸及激动剂处理细胞,采用流式细胞仪及Western印迹技术检测细胞中ICAM-1及VCAM-1蛋白质的表达.结果 反义阻断PPARγ在mRNA水平上的表达,ECV-304细胞ICAM-1及VCAM-1的表达上调,而PPARγ激动剂Ciglitizone则降低ICAM-1及VCAM-1的表达.结论 PPARγ激活可负调控AGEs诱导的血管内皮细胞ICAM-1和VCAM-1的表达,这是PPARγ抑制糖尿病血管并发症及动脉粥样硬化发生发展的可能机制之一.     

糖基化终产物对人血管内皮细胞基质金属蛋白酶-2表达和活性的影响

目的探讨糖基化终产物(AGEs)对人血管内皮细胞ECV304基质金属蛋白酶-2(MMP-2)表达和活性的影响。方法应用AGEs作用ECV304细胞后,采用RT-PCR法及明胶酶图分析方法分析MMP-2表达和活性与AGEs浓度和时间的关系。结果 RT-PCR结果显示,AGEs能够诱导ECV304细胞表达MMP-2,其诱导表达具有浓度和时间依赖性(P<0.01)。明胶酶图分析显示,MMP-2降解明胶的活性与AGEs具有浓度依赖性。结论 AGEs能够促进ECV304细胞MMP-2的表达和活性,推测AGEs调控MMP-2表达和活性可能在血管壁内皮细胞影响动脉粥样硬化的进展中发挥一定的作用。     

糖基化终末产物对ECV-304细胞ICAM-1及VCAM-1表达的影响

目的探讨糖基化终产物(AGEs)与其受体(RAGE)对ECV-304细胞上血管细胞黏附分子-(1ICAM-1)和细胞间黏附分子-(1VCAM-1)的影响,为研究AGEs在动脉粥样硬化发生发展中的作用提供实验依据。方法采用体外制备的糖基化终产物——糖基化的牛血清白蛋白(AGE-BSA)和反义RAGE寡核苷酸处理ECV-304细胞,通过流式细胞仪技术、Western印迹技术检测ECV-304细胞上ICAM-1、VCAM-1蛋白质的表达。结果AGEs处理ECV-304细胞后,可诱导ECV-304细胞上VCAM-1和ICAM-1的表达增加,且这种效应呈剂量依赖性。采用反义技术阻断RAGE的mRNA水平的翻译后,ECV-304细胞ICAM-1和VCAM-1的表达呈剂量依赖性降低。结论AGEs可通过与其膜受体RAGE的结合激活血管内皮细胞VCAM-1和ICAM-1的表达,这可能是AGEs促动脉粥样硬化发生发展的机制之一。     

血管紧张素受体1阻断剂替米沙坦、厄贝沙坦具有激活PPARα的作用

目的 通过观察替米沙坦、厄贝沙坦对PPARa转录活性的影响以探讨其改善糖脂代谢的机制.方法 将构建的PPARa表达质粒、PPAR反应元件(PPRE)调控的荧光素酶表达质粒与pRL表达载体以脂质体(SuperFect)瞬时共转染COS-7细胞后,分别加入不同浓度的替米沙坦及厄贝沙坦,继续培养细胞不同时间,用双荧光索酶基因报告系统检测荧光素酶活性以反映PPARa转录活性.不同浓度的厄贝沙坦及替米沙坦孵育3T3-L1脂肪细胞,分别应用RT-PCR和Western印迹测定细胞的PPARα mRNA和蛋白表达水平.结果 (1)替米沙坦和厄贝沙坦均呈剂量和时间依赖性地增加COS-7细胞的PPARα转录活性,在60 h作用均达高峰,两者在100μmol/L浓度时PPRE调控的荧光素酶活性较对照组增高3.8倍和2.6倍(均P<0.01);(2)替米沙坦及厄贝沙坦激活PPARα的作用不能被PPARγ脚特异性抑制剂GW9662抑制;(3)替米沙坦和厄贝沙坦呈剂量依赖性增加3T3-L1脂肪细胞PPARα mRNA和蛋白表达水平.结论 血管紧张素受体阻断剂替米沙坦和厄贝沙坦增加PPARα转录活性,可能通过此途径改善糖脂代谢.     

激活过氧化物酶增殖物激活受体对血管紧张素Ⅱ诱导内皮细胞中单核细胞趋化因子-1表达的影响

目的探讨血管紧张素Ⅱ（AngⅡ）对人脐静脉内皮细胞分泌单核细胞趋化因子1（MCP-1）的影响，以及激活过氧化物酶增殖物激活受体（PPARs）中α，γ亚型对MCP-1的影响。方法以RT—PCR和ELISA方法测定AngⅡ在不同浓度（10^-8～10^-6mol／L）对人脐静脉内皮分泌MCP-1的影响以及血管紧张素Ⅱ1型受体（AT1R）拮抗剂缬沙坦的干预作用；同时分析PPARa配体非诺贝特及PPAR7配体罗格列酮的干预对AngⅡ诱导MCP-1表达的效应。结果 AngⅡ显著刺激人脐静脉内皮细胞系（HUVEC-12）表达MCP1，增加的程度与AngⅡ的浓度呈正相关，罗格列酮和非诺贝特均可呈浓度依赖的抑制HUVEC12中由AngⅡ所诱导的MCP-1的表达。结论 AngⅡ可刺激HUVEC-12细胞表达MCP-1，其作用与AT1R受体相关；激动剂PPARa、PPAR7均可明晁加制AngⅡ所诱导的内古纲晌巾MCP-1嘉亍大     

糖基化终末产物诱导大鼠视网膜微血管内皮细胞表型和功能改变

目的 研究糖基化终末产物(AGEs)诱导大鼠视网膜微血管内皮细胞表型和功能的变化及相关机制.方法 SPF级雄性Wistar大鼠20只,7～8周龄,体重260～280 g,分离培养大鼠视网膜内皮细胞,采用免疫荧光染色、流式细胞术和体外形成毛细血管样网络结构的方法进行鉴定.视网膜内皮细胞在AGEs刺激后,用噻唑蓝(MTT)法分析细胞的增殖能力;用膜连蛋白V/碘化丙啶(Annexin V/PI)双染法检测细胞凋亡;采用逆转录聚合酶链反应(RT-PCR)和流式细胞术检测细胞AGEs受体、人蛋白激酶C(PKC)、细胞间黏附分子1(ICAM-1)和诱导型一氧化氮合酶(iNOS)的表达变化.采用t检验进行统计学分析.结果 分离纯化的大鼠视网膜内皮细胞表达血管性血友病因子(vWF)并在人工基膜上形成毛细血管网络结构.AGEs以时间和剂量依赖的方式抑制大鼠视网膜内皮细胞增殖能力,在200 mg/L的AGEs组培养至第5天时细胞增殖能力低于对照组(t=8.9,P＜0.05),7 d和9 d时抑制作用更明显(t值分别为15.7和46.1,均P＜0.01).400 mg/L AGEs组在第3天开始出现细胞增殖减慢(t=12.5,P＜0.05),从第5天开始增殖速度明显低于对照组(t值分别为22.4、41.5和77.7,均P＜0.01).并且AGEs诱导视网膜内皮细胞凋亡.进一步分析发现AGEs上调了大鼠视网膜内皮细胞AGEs受体、PKC、ICAM-1和iNOS的mRNA表达(t值分别为91.8、9.22、16和42,均P＜0.01)和蛋白水平的表达(t值分别为20.2、12.3、7.7和13.9,均P＜0.01).结论 AGEs可能通过上调AGEs受体诱导大鼠视网膜微血管内皮细胞表型和功能的改变.     

糖基化终产物对人脐静脉内皮细胞血管内皮生长因子mRNA及蛋白表达的影响

目的研究糖基化终产物（AGEs）对人脐静脉山皮细胞血管内皮生长凼子（VEGF）mRNA及蛋白表达的影响，探讨AGEs在糖尿病动脉粥样硬化中的作用。方法将人脐静脉内皮细胞株ECV304用BSA及小同浓度的AGEs孵育24h及400mg／L的AGEs孵育0、12、24及36h，采用原位杂交及Westem blot方法检测VEGFmRNA及蛋白的表达。结果人脐静脉内皮细胞在100、200及400mg／LAGEs孵育24h后，VEGFmRNA及蛋白表达均显著高于BSA对照组（P〈0．05），在用400mg／LAGEs分别孵育12．24及36h后，各组内皮细胞VEGF mRNA及蛋白表达量也明显高于0h对照组（P〈O．05）。结论AGEs可增加人脐静脉内皮细胞VEGF mRNA及蛋白的表达，且与浓度和时间呈正相关。     

脂肪酸影响血管内皮细胞纤溶酶原激活物抑制剂-1表达的机制初探

目的 探讨脂肪酸影响血管内皮细胞纤溶酶原激活物抑制剂－1 机制。方法 以PAI－1启动子控制表达氯霉素转移乙酰酶（CAT）报告基因的重组质粒－PAI－pCAT转染人血管内皮细胞株ECV304，并且部分共转染不同量的过氧化增殖物激活型肥体（PPAR）α或PPARγ表达载体。分别以亚麻酸，亚油酸，油酸，硬脂酸诱导转染细胞，酶联免疫吸附CAT表达量显示启动子片面转录活性。结果 亚麻酸，亚油酸，油酸诱导以及增加PPARα表达可提高PAI－1启动子转录活性，硬脂酸诱导和增加PPARγ表达无影响。结论 不饱和脂肪酸可通过提高PAI－1转录活性诱导其在血管内皮细胞的表达，此作用涉及PPARγα对PAI-1基因转录的诱导调节。     

过氧化物酶体增殖物激活受体γ对糖基化终产物诱导大鼠血管平滑肌细胞增殖的作用

目的观察吡格列酮对糖基化终产物（AGEs）刺激下大鼠血管平滑肌细胞（VSMCs）增殖的作用及对过氧化物酶体增殖物激活受体γ（PPARγ）基因及蛋白表达水平的影响,探讨PPARγ在AGEs诱导VSMCs增殖中的作用。方法（1）MTY法观察不同浓度、不同时间的AGEs对VSMCs增殖的影响及吡格列酮（1．0、10、100μmol／L）与AGEs共孵育对VSMCs增殖的干预作用。（2）用半定量逆转录聚合酶链反应测定VSMCs中PPARγ mRNA的表达。（3）用Western blot法检测PPARγ的蛋白表达。结果AGEs作用导致VSMCs增殖,AGEs抑制PPARγ mRNA和蛋白表达水平,这种抑制作用随着AGEs干预的时间延长和浓度的增加而增强（P〈0．05）。PPARγ激活剂吡格列酮通过增加PPARγ的表达,抑制AGEs诱导的VSMCs增殖。结论PPARγ表达的下降可能是糖尿病易患动脉粥样硬化的重要原因之一。     

替米沙坦对人血管内皮细胞炎性反应的影响及其相关机制

目的探讨替米沙坦对晚期糖基化终末产物(AGEs)诱导人脐静脉内皮细胞(HUVECs)炎性反应的影响及与过氧化物酶体增殖物激活受体γ(PPARγ)的关系。方法酶消化法获取HUVECs。实验分8组:BSA组;AGEs组;以不同浓度替米沙坦刺激细胞30 min,再加AGEs,依次为TM 1组(1 nmol/L)、TM 2组(10 nmol/L)、TM 3组(100 nmol/L)、TM 4组(1000 nmol/L);GW9662组:GW9662预先刺激细胞30 min后,加替米沙坦和AGEs;15d-PGJ_2组:15d-PGJ_2预先刺激细胞30 min后,加AGEs。各组均刺激细胞24 h。采用RT-PCR法检测PPARγ、AGEs受体(RAGE)和单核细胞趋化蛋白1(MCP-1)mRNA的表达;荧光探针2,7-二氯二氢荧光素乙酰乙酸检测细胞内活性氧的变化。结果替米沙坦呈浓度依赖性抑制AGEs诱导的MCP-1 mRNA表达。与AGEs组比较,TM 4组和15d-PGJ_2组PPARγ mRNA水平明显升高,RAGE mRNA和MCP-1 mRNA水平明显下降,活性氧荧光信号强度明显减弱。与TM 4组比较,GW9662组PPARγ mRNA的表达水平明显下降,RAGE mRNA和MCP-1 mRNA的表达水平明显升高,活性氧荧光信号强度明显增强。结论替米沙坦可抑制AGEs诱导的HUVECs炎性反应;替米沙坦可能通过激活PPARγ抑制HUVECs炎性反应。     

胰岛素瘤的解剖分布特点分析

目的胰岛素瘤是最常见的胰腺神经内分泌肿瘤，因其临床表现多样，导致诊断困难。影像学诊断尤其是超声内镜(EUS)在胰岛素瘤的诊断中起着重要作用，拥有较高的敏感性和特异性。本研究拟通过明确胰岛素瘤的解剖分布特点，以期有助于提高影像学的诊断准确率和降低漏诊率，尤其是在教育和培训实践中对于EUS的学习者更具有指导价值。 方法回顾性分析解放军总医院第一医学中心病案资料数据库1993年1月至2019年11月经外科手术、病理确诊为胰岛素瘤的患者的临床资料，检索方法采取搜索术后病理诊断为"胰岛素瘤"的病例，通过查阅病例的方法，提取出胰岛素瘤的大小和解剖分布等数据，进一步分析其特点。 结果共检索到确诊为胰岛素瘤的患者116例，其中，男45例、女71例，年龄13～76岁，平均年龄(44.4±14.85)岁。胰岛素瘤单发110例(94.8%)、多发6例(5.2%)。位置分布：头颈部46例(39.7%)，单发45例、多发1例；体尾部68例(58.6%)，单发65例、多发3例；全胰腺多发2例(1.7%)。病变大小特点：最大径0.4～3.4 cm，平均大小(1.53±0.58)cm。≤1 cm 29例、>1 cm而≤1.5 cm41例、>1.5 cm而≤2.0 cm28例，≤3 cm 15例，>3 cm 3例。年龄与肿瘤的大小相关，≤44岁患者肿瘤平均大小为(1.36±0.51)cm、>44岁患者肿瘤平均大小为(1.70±0.60)cm，P<0.05。头颈部的肿瘤大于体尾部的肿瘤，头颈部肿瘤平均大小(1.66±0.63)cm，体尾部(1.42±0.52)cm，P<0.05。 结论胰岛素瘤在胰腺体尾部较头颈部更好发；绝大多数单发，但可以全胰腺多发；多数小于1.5 cm，肿瘤的大小与患者年龄和肿瘤的解剖分布相关。     

Pathogenesis of carcinoma of the papilla of Vater

Most adenomas and carcinomas of the small intestine and extrahepatic bile ducts arise in the region of the papilla of Vater. In familial adenomatous polyposis (FAP) it is the main location for carcinomas after proctocolectomy. In many cases symptoms due to stenosis lead to diagnosis at an early tumor stage. In about 80%, curative intended resection is possible. Operability is the most relevant prognostic factor. Most ampullary carcinomas resp. carcinomas of the papilla of Vater develop from adenomatous or flat dysplastic precursor lesions. They can be sited in the ampulloduodenal part of the papilla of Vater, which is lined by intestinal mucosa. They also can develop in deeper parts of the ampulla, which are lined by pancreaticobiliary duct mucosa. Intestinal-type adenocarcinoma and pancreaticobiliary-type adenocarcinoma represent the main histological types of ampullary carcinoma. Furthermore, there exist unusual types and undifferentiated carcinomas. Many carcinomas of intestinal type express the immunohistochemical marker profile of intestinal mucosa (keratin 7?, keratin 20+, MUC2+). Carcinomas of pancreaticobiliary type usually show the immunohistochemical profile of pancreaticobiliary duct mucosa (keratin 7+, keratin 20?, MUC2?). Even poorly differentiated carcinomas, as well as unusual histological types, may conserve the marker profile of the mucosa they developed from. These findings underline the concept of histogenetically different carcinomas of the papilla of Vater which develop either from intestinal- or from pancreaticobiliary-type mucosa of the papilla of Vater. Molecular alterations in ampullary carcinomas are similar to those of colorectal as well as pancreatic carcinomas, although they appear at different frequencies. In future studies, molecular alterations in ampullary carcinomas should be correlated closely with the different histologic tumor types. Consequently, the histologic classification should reflect the histogenesis of ampullary tumors from the two different types of papillary mucosa.     

Demonstration of the mechanism of action of biguanides

Summary Palmitic acid oxidation in rat diaphragm homogenate is depressed by biguanide concentrations that are still incapable of inhibiting oxidative phosphorylation. Glucose oxidation is not directly effected by the same biguanide concentrations: however, the inhibitory effect of palmitic acid on glucose oxidation is partly removed by biguanides. Inhibition of fatty acid oxidation, which accounts for most of the metabolic effects caused by these drugs, can be regarded as the fundamental mechanism of action of biguanides. There is some evidence suggesting that these drugs might interact with carnitine, thus preventing long-chain fatty acids from being transported across the mitochondrial membrane to the site of oxidation. Traduzione a cura degli AA.     

Lack of correlation of degree of villous atrophy with severity of clinical presentation of coeliac disease

BACKGROUND AND AIM: Both the clinical presentation and the degree of mucosal damage in coeliac disease vary greatly. In view of conflicting information as to whether the mode of presentation correlates with the degree of villous atrophy, we reviewed a large cohort of patients with coeliac disease. PATIENTS AND METHODS: We correlated mode of presentation (classical, diarrhoea predominant or atypical/silent) with histology of duodenal biopsies and examined their trends over time. RESULTS: The cohort consisted of 499 adults, mean age 44.1 years, 68% females. The majority had silent coeliac disease (56%) and total villous atrophy (65%). There was no correlation of mode of presentation with the degree of villous atrophy (p=0.25). Sixty-eight percent of females and 58% of males had a severe villous atrophy (p=0.052). There was a significant trend over time for a greater proportion of patients presenting as atypical/silent coeliac disease and having partial villous atrophy, though the majority still had total villous atrophy. CONCLUSIONS: Among our patients the degree of villous atrophy in duodenal biopsies did not correlate with the mode of presentation, indicating that factors other than the degree of villous atrophy must account for diarrhoea in coeliac disease.     

血吸虫童虫生长发育及其机制的研究进展

血吸虫童虫是宿主免疫系统攻击的重要靶标,包括皮肤型、肺型和肝门型童虫。宿主分子对童虫生长发育具有重要作用。童虫生长发育机制包括免疫调节、信号转导、性别发育及凋亡等。肌动蛋白、组织蛋白酶、烯醇化酶和葡萄糖基转移酶等分子为血吸虫童虫生长发育的重要分子。本文对血吸虫童虫生长发育及其机制的研究进展做一综述。     

124例耐多药结核分枝杆菌基因突变特征分析

目的对临床分离的耐多药结核分枝杆菌相关基因的突变特征进行分析。方法对124例耐多药结核分枝杆菌以及50株敏感株的耐药相关基因(包括异烟肼inh A、kat G、oxyR-ahp C间隔区以及利福平rpo B)进行序列测定,分析其基因突变情况。结果异烟肼耐药inh A基因突变率为14.5%;kat G基因突变率为70.2%(87/124),主要位于315位;oxyR-ahp C间隔区突变率为15.3%;inh A、kat G两种基因同时突变率75.0%,三种基因同时突变率为89.5%。利福平rpo B基因突变的检出率高达95.2%,突变主要发生在531、526、516位点。结论我省耐多药菌异烟肼耐药相关基因最常见突变为kat G 315、inh A C-T(-15)、axyR-ahp C间隔区(-10)C-T,利福平为rpo B531、526、516。结合MDR-TB耐药相关基因的特征分析,可以建立一种快速、准确、特异的适合于我省的检测结核菌耐多药性的新方法。     

氯硝柳胺悬浮剂的毒性评价

目的评价氯硝柳胺悬浮剂的毒性，为现场大规模应用灭螺提供依据。方法按照中华人民共和国国家标准GB 15670-1995《农药登记毒理学试验方法》和鱼类毒性试验方法进行。结果经口、经皮肤的LDso雌、雄性大鼠均>5 000 mg/kg，经呼吸道的LCso雌、雄性大鼠均>5 000mg/m3，该药经口、经皮肤、经呼吸道毒性均属微毒类药物；兔眼用药后，观察期内无不良反应，对眼无刺激性；皮肤用药后对皮肤无刺激性。与氯硝柳胺原药、氯硝柳胺乙醇胺盐原药和氯硝柳胺乙醇胺盐可湿性粉剂相比，氯硝柳胺悬浮剂对鱼急性毒性最低。结论氯硝柳胺悬浮剂属微毒类药物，对鱼的毒性低于其乙醇胺盐可湿性粉剂，适合于现场应用。     

Assessment of quality of life of parents of children with juvenile chronic arthritis

The aim of the study was to assess the quality of life (QOL) and the psychological status of parents of children with juvenile chronic arthritis (JCA). The QOL, anxiety and depression of the parents of 28 children with JCA were evaluated and compared to those of the parents of 28 healthy children. Mothers of JCA children and mothers of healthy children reported similar QOL. The reported anxiety and depression levels were similar for mothers and fathers in both groups. The parents of children with pauciarticular-type JCA reported lower QOL and higher levels of anxiety and depression than the parents of children with other types, namely polyarticular and systemic JCA. These findings may be explained by the fact that the pauciarticular patients had shorter disease duration and were less frequently seen in the outpatient clinic. The QOL of mothers of children with JCA was found to be slightly impaired in the group of children with pauciarticular JCA. Future larger studies are needed to confirm these results, as the number of subjects in the three groups was rather low. Received: 26 September 2001 / Accepted: 8 February 2002     

Numerical Simulation of the Effect of Rate of Change of Glucose on Measurement Error of Continuous Glucose Monitors

Background

A 5-day in-patient study designed to assess the accuracy of the FreeStyle Navigator® Continuous Glucose Monitoring System revealed that the level of accuracy of the continuous sensor measurements was dependent on the rate of glucose change. When the absolute rate of change was less than 1 mg•dl⁻¹•min⁻¹ (75% of the time), the median absolute relative difference (ARD) was 8.5%, with 85% of all points falling within the A zone of the Clarke error grid. When the absolute rate of change was greater than 2 mg•dl⁻¹•min⁻¹ (8% of the time), the median ARD was 17.5%, with 59% of all points falling within the Clarke A zone.

Method

Numerical simulations were performed to investigate effects of the rate of change of glucose on sensor measurement error. This approach enabled physiologically relevant distributions of glucose values to be reordered to explore the effect of different glucose rate-of-change distributions on apparent sensor accuracy.

Results

The physiological lag between blood and interstitial fluid glucose levels is sufficient to account for the observed difference in sensor accuracy between periods of stable glucose and periods of rapidly changing glucose.

Conclusions

The role of physiological lag on the apparent decrease in sensor accuracy at high glucose rates of change has implications for clinical study design, regulatory review of continuous glucose sensors, and development of performance standards for this new technology. This work demonstrates the difficulty in comparing accuracy measures between different clinical studies and highlights the need for studies to include both relevant glucose distributions and relevant glucose rate-of-change distributions.     

治疗高血压药物的经济学评价

重视高血压治疗中的经济学评价,对利用我国有限的卫生资源来遏制高血压对人民群众的危害有着重要的现实意义。药物经济学对于药物治疗的成本和治疗的结果给予同样的关注。因为治疗高血压的费用,不仅涉及药物价格,还包括患者的危险水平,降压疗效和对临床终点事件的影响,以及治疗的依从性和安全性。因此药物经济学更强调整体成本和价-效比。低危病人,若非药价低廉,治疗的价-效比不够理想。而在高危的患者,价-效比越小越经济而不是药费越便宜越好。