Oxidized LDL downregulates ATP-binding cassette transporter-1 in human vascular endothelial cells via inhibiting liver X receptor (LXR)

OBJECTIVE: ATP-binding cassette transporter-1 (ABCA1) mediates the active efflux of cholesterol and phospholipids, playing an important role in cholesterol homeostasis and atherogenesis. Oxidized low density lipoprotein (oxLDL) is an atherogenic molecule associated with the vascular endothelial dysfunction and development of atherosclerotic plaque. This report describes the effect of copper-catalyzed oxLDL on the regulation of ABCA1 in human endothelial cells (ECs). METHODS AND RESULTS: oxLDL downregulated ABCA1 at both mRNA and protein levels in a dose-dependent manner. This inhibitory effect of oxLDL was observed with both minimally and extensively oxLDL. Transfection of the ABCA1 promoter luciferase revealed oxLDL to substantially decrease ABCA1 promoter activity at basal conditions and after stimulation by overexpressing the liver X receptor LXRalpha and retinoid X receptor RXRalpha. oxLDL also attenuated LXR activation by blocking LXR ligand binding and interfering with the generation of 27-hydroxycholesterol, an LXR endogenous ligand. Furthermore, oxLDL inhibited exogenous cholesterol- and oxysterol-induced endothelial ABCA1 induction. CONCLUSION: oxLDL downregulated ABCA1 by inhibiting LXR activation in endothelial cells. Such an effect may contribute to endothelial dysfunction and plaque formation.     

Genetic alterations of hepatocellular carcinoma by random amplified polymorphic DNA analysis and cloning sequencing of tumor differential DNA fragment

AIM: To study the genetic alterations and their association with clinicopathological characteristics of hepatocellular carcinoma (HCC), and to find the tumor related DNA fragments. METHODS: DNA isolated from tumors and corresponding noncancerous liver tissues of 56 HCC patients was amplified by random amplified polymorphic DNA (RAPD) with 10 random 10-mer arbitrary primers. The RAPD bands showing obvious differences in tumor tissue DNA corresponding to that of normal tissue were separated, purified, cloned and sequenced. DNA sequences were analyzed and compared with GenBank data. RESULTS: A total of 56 cases of HCC were demonstrated to have genetic alterations, which were detected by at least one primer. The detestability of genetic alterations ranged from 20% to 70% in each case, and 17.9% to 50% in each primer. Serum HBV infection, tumor size, histological grade, tumor capsule, as well as tumor intrahepatic metastasis, might be correlated with genetic alterations on certain primers. A band with a higher intensity of 480 bp or so amplified fragments in tumor DNA relative to normal DNA could be seen in 27 of 56 tumor samples using primer 4. Sequence analysis of these fragments showed 91% homology with Homo sapiens double homeobox protein DUX10 gene. CONCLUSION: Genetic alterations are a frequent event in HCC, and tumor related DNA fragments have been found in this study, which may be associated with hepatocarcin-ogenesis. RAPD is an effective method for the identification and analysis of genetic alterations in HCC, and may provide new information for further evaluating the molecular mechanism of hepatocarcinogenesis.     

Histopathology and TB-PCR kit analysis in differentiating the diagnosis of intestinal tuberculosis and Crohn��s disease

AIM: To compare the histopathologic features of intestinal tuberculosis (ITB) and Crohn’s disease (CD) and to identify whether polymerase chain reaction for Mycobacterium tuberculosis (TB-PCR) would be helpful for differential diagnosis between ITB and CD.METHODS: We selected 97 patients with established diagnoses (55 cases of ITB and 42 cases of CD) who underwent colonoscopic biopsies. Microscopic features of ITB and CD were reviewed, and eight pathologic parameters were evaluated. Nine cases of acid fast bacilli culture-positive specimens and 10 normal colonic tissue specimens were evaluated as the positive and negative control of the TB-PCR test, respectively. PCR assays were done using two commercial kits: kit detected IS6110 and MPB64, and kit detected IS6110 only; a manual in-house PCR method was also performed on formalin-fixed, paraffin-embedded colonoscopic biopsy specimens.RESULTS: Statistically significant differences were noted between ITB and CD with regard histopathologic criteria: size of granulomas (P = 0.000), giant cells (P = 0.015), caseation necrosis (P = 0.003), confluent granulomas (P = 0.001), discrete granulomas (P = 0.000), and granulomas with lymphoid cuffs (P = 0.037). However, 29 cases (52.7%) of ITB showed less than five kinds of pathologic parameters, resulting in confusion with CD. The sensitivities and specificities of the TB-PCR test by kit , kit , and the in-house PCR method were 88.9% and 100%, 88.9% and 100%, and 66.7% and 100% in positive and negative controls, respectively. The PCR test done on endoscopic biopsy specimens of ITB and CD were significantly different with kit (P = 0.000) and kit (P = 0.000). The sensitivities and specificities of TB-PCR were 45.5% and 88.1%, 36.4% and 100%, and 5.8% and 100%, for kit and kit and in-house PCR method on endoscopic biopsy specimens. Among the 29 cases of histopathologically confusing CD, 10 cases assayed using kit and 6 cases assayed using kit were TB-PCR positive. A combination of histologic findings and TB-PCR testing led to an increase of diagnostic sensitivity and the increase (from 47.3% to 58.2) was statistically significant with kit (P = 0.000).CONCLUSION: The TB-PCR test combined with histopathologic factors appears to be a helpful technique in formulating the differential diagnosis of ITB and CD in endoscopic biopsy samples.     

应用吊瓶式输液器拆分输注甘露醇

用一次性使用吊瓶式输液器准确拆分全量甘露醇包装液,代替普通输液器给患者输注甘露醇。结果一次性使用吊瓶式输液器能利用全量甘露醇包装液准确拆分输注不同剂量的甘露醇,减轻了护士工作量并使输注的甘露醇剂量准确,杜绝了由于超剂量使用甘露醇导致的不良反应,无1例发生医疗纠纷。提示使用一次性吊瓶式输液器拆分输注甘露醇原包装液,能确保应用甘露醇剂量的准确性。     

含H5N1禽流感病毒核酸序列的病毒样颗粒的制备及应用

目的构建耐RNase酶的内含禽流感病毒H5N1亚型部分核酸序列的病毒样颗粒。方法构建中间载体pET32a-MS2,将禽流感病毒H5N1亚型的HA、NA和M基因片断连接到中间载体上,构建原核表达载体pET32a-MS2-HA,pET32a-MS2-NA和pET32a-MS2-M,分别转化宿主菌,诱导表达制备病毒样颗粒。结果表达载体经PCR、酶切鉴定和测序分析后证实构建成功。电镜观察到了病毒样颗粒,荧光定量RT-PCR试验表明颗粒稳定性良好。结论成功构建了含禽流感病毒H5N1部分核酸序列的病毒样颗粒且稳定性良好,可作为禽流感H5N1亚型RNA检测的标准品和质控品。     

2008年黑龙江省安达市改水降氟工程运行现状调查分析

目的 了解2008年黑龙江省安达市降氟改水工程分布、运行现状和存在问题.方法 对安达市全部降氟改水工程进行全球定位系统(GPS)定位及工程运行和管理现状逐项调查.结果 共调查降氟改水工程317处,由于改水工程超期使用、管理不善,停用工程51处,占16.09%,报废工程77处,占24.29%;供水设备未配套、供水管网未铺造成工程闲置83处,占26.18%;目前运行的改水工程106处,占33.44%,水氟达标的工程只有46处,占14.51%,受益人口数3.6万人.结论 安达市降氟改水工程运行状况令人十分担忧,报废、停用、闲置现象严重,急需改进.     

陕西省世界银行贷款/英国赠款结核病控制项目6年效果分析

目的分析陕西省世界银行贷款/英国赠款结核病控制项目实施6年的效果,为我省结核病防治工作可持续发展提供可参考。方法收集2002—2007年陕西省107个区(县)项目季报表,对全省DOTS策略覆盖率、肺结核可疑症状者就诊率、肺结核患者登记率和治愈率等主要指标进行统计分析。结果全省2002年启动DOTS策略,2005年以县为单位DOTS策略覆盖率达到了100%。2002—2007年,全省接诊肺结核可疑症状者386281例,可疑症状者就诊率逐年增高;发现涂阳肺结核患者61293例,其中新涂阳肺结核患者45123例,新涂阳肺结核登记率平均为20.45/10万,2004—2007年,新涂阳肺结核发现率平均为87.72%。初治涂阳肺结核治愈率平均为90.6%,复治涂阳患者治愈率平均为81.7%。结论该项目促进了DOTS策略的快速扩展,提高了肺结核患者的发现率和治愈率,产生了明显的社会效益和经济效益。     

帕金森病患者非运动症状的发生状况及影响因素

目的 研究帕金森病(PD)非运动症状(non-motor symptoms,NMS)的各项发生率、发生数及可能影响因素.方法 对131例门诊确诊为PD的患者进行NMS问卷(NMSQuest)调查,研究各项发生率、总发生数的状况及其与各种可能因素间的关系.结果所有入组PD患者均有NMS,分布于PD的各个阶段,平均每例患者有11项NMS;各项发生率中以自主神经症状如夜尿、便秘等的发生率最高,入组年龄≥65岁患者组在嗅觉减退、吞咽困难和便秘等多项发生率上高于＜65岁组,强直-少动型患者在抑郁相关项目上发生率高于震颤型患者;NMS发生数与病程、患者入组年龄、统一帕金森病评定量表(UPDRS)总分和Hoehn-Yahr(H-Y)分级呈正相关.结论 NMS普遍存在于各期的PD患者,各项发生率随PD患者的临床特点不同而变化,NMS发生数与PD的疾病严重程度呈正相关,应提高对NMS诊断和合理治疗的重视.     

鞘磷脂酶在柯萨奇病毒B3感染心肌细胞中的作用

目的检测柯萨奇病毒B3(CVB3)感染心肌细胞后鞘磷脂酶活性的变化、鞘磷脂酶mRNA、CVB3RNA的表达、病毒滴度的变化及心肌细胞表型的变化。初步探讨鞘磷脂酶在CVB3感染的心肌细胞中的作用。方法 CVB3感染心肌细胞后不同时间点,用薄层色谱法(TLC)检测鞘磷脂酶活性;用Real time-PCR检测心肌细胞中鞘磷脂酶mRNA与CVB3RNA的表达;同时取细胞培养上清液检测CVB3滴度。用Annexin V-FITC,PI双染法观察心肌细胞表型变化。设正常心肌细胞对照组,每个时间点设三个复孔。结果病毒感染组与正常心肌细胞相比,酸性鞘磷脂酶在4h、12h时活性有升高,其mRNA表达在2h时有升高趋势(P0.05);中性Mg2+非依赖性鞘磷脂酶活性在4h时有明显升高,在12h时活性下降,而其mRNA表达在20h时升高(P0.05);中性Mg2+依赖性鞘磷脂酶活性于病毒感染后4h、12h无明显变化,其mRNA表达在各组间也无明显变化。心肌细胞内病毒RNA表达在4h时有升高趋势,8h后开始明显升高(P0.01)。CVB3病毒滴度在2h时明显升高(P0.05)。心肌细胞在病毒感染2h后早期凋亡及坏死开始增加,4h时凋亡坏死增加更明显,到24h时坏死心肌细胞明显增多。以上实验均重复3次。结论中性Mg2+非依赖性鞘磷脂酶与酸性鞘磷脂酶在CVB3侵入心肌细胞及病毒的扩散增殖过程中可能起重要作用;而中性Mg2+依赖性鞘磷脂酶在此过程中可能不起作用。     

动脉硬化性闭塞症坏死期保守治疗适应指标观察

目的探讨动脉硬化性闭塞症坏死期保守治疗预后与治疗前血小板、球蛋白、C反应蛋白、凝血四项、血液流变学、踝肱指数的关系,为患者选择保守治疗提供依据。方法根据中医内、外治法及基础治疗的疗效,将150例患者分为愈显组59例、好转组71例、未愈组20例。对3组患者治疗前的血小板、球蛋白、C反应蛋白、凝血四项、血液流变学、踝肱指数检测数据进行统计学分析。结果 3组患者治疗前全血低切黏度差异有统计学意义（χ2=15.99）,两两组间比较差异有统计学意义（P〈0.01）。3组踝肱指数差异有统计学意义（χ2=8.56）,两两组间比较差异有统计学意义（P〈0.01）。3组纤维蛋白原、C反应蛋白均明显高于正常值。结论动脉硬化性闭塞症坏死期患者全血低切黏度、踝肱指数值可以作为选择保守治疗的参考指标。