三种斑蝥素盐类物质对肝癌QGY-7703细胞生长的抑制作用

目的 考察斑蝥素酸镁、斑蝥素酸钠及斑蝥素酸钾,三种斑蝥素的盐类物质对肝癌QGY-7703细胞增殖的抑制作用.方法 采用WST-1比色法测定三种斑蝥素盐类物质对肝癌QGY-7703细胞的生长抑制率.结果 三种斑蝥素盐类物质对肝癌QGY-7703细胞的生长具有显著的抑制作用,抑制率随药物浓度升高其抑制作用增强,呈剂量效应关系.其半数抑制率IC50分别为9.48、33.80、35.04 μmol/L.结论 斑蝥素酸镁对肝癌QGY-7703细胞的抑制作用明显好于斑蝥素酸钠和斑蝥素酸钾,具有开发潜力.     

5-Aza-CdR对肝癌细胞DAPK基因甲基化的影响

目的探讨去甲基化药物5-氮杂-2’-脱氧胞苷(5-Aza-CdR)对人肝癌细胞株SMMC-7721中死亡相关蛋白激酶(DAPK)基因启动子CpG岛甲基化的影响。方法不同浓度(0.5μmol/L、5.0μmol/L、50.0μmol/L)5-Aza-CdR处理人肝癌细胞株SMMC-772172小时后,用焦磷酸测序法检测处理前后DAPK基因启动子CpG岛甲基化水平。以未经处理的人肝癌细胞株SMMC-7721作为对照组。结果未经5-Aza-CdR处理的肝癌细胞SMMC-7721细胞株DAPK基因启动子高度甲基化,平均水平为76.71%。经3种不同浓度药物处理72小时后,各组间甲基化平均水平分别为78.29%、77.57%和66.00%,各组间甲基化水平差异均有统计学意义(F=39.71,P<0.01)。其中,50.0μmol/L处理组细胞甲基化水平最低,与其他各组差异均有统计学意义(P<0.01)。结论 SMMC-7721细胞株DAPK基因启动子区呈高度甲基化状态;5-Aza-CdR能够逆转人肝癌细胞SMMC-7721DAPK基因启动子区域的甲基化状态。     

内质网应激-自噬肝癌SMMC-7721细胞模型的复制及金刚藤的干预

背景内质网应激(endoplasmic reticulum stress, ERS)-自噬是肝癌细胞一种自我保护性机制.内质网(endoplasmic reticulum, ER)应激诱导剂二硫苏糖醇(dithiothreitol,DTT)能诱导肝癌细胞ERS-自噬.课题组前期研究证实金刚藤具有抑制肝癌细胞的作用,那么肝癌细胞ERS-自噬是否影响金刚藤抑制肝癌细胞的作用尚不清楚.目的探讨ERS-自噬肝癌SMMC-7721细胞模型的复制及金刚藤的干预.方法ERS-自噬肝癌SMMC-7721细胞模型的复制:采用不同剂量的DTT(50、200、500μmol/L)处理SMMC-7721细胞24 h,设DTT 0μmol/L组为control组,透射电镜观察各模型组细胞自噬小体,运用western blot法测定各组细胞微管相关蛋白1轻链3-Ⅱ(microtubuleassociated protein 1 light chain 3-Ⅱ, LC3-Ⅱ)蛋白的表达.金刚藤对ERS-自噬肝癌SMMC-7721细胞存活率的影响:将SMM C-7721细胞分为空白组(control组)、金刚藤组(JGT组)、DTT+JGT组, control、JGT组分别给予空白血清、金刚藤含药血清处理, DTT+JGT组予DTT构建的ERS模型后给予金刚藤含药血清处理,采用CCK-8法检测细胞的存活率.结果 DTT(50、200、500μmol/L)组细胞的细胞质内均可见自噬小体,随着DTT浓度的增加,自噬小体数量逐渐增加, DTT 500μmol/L组可见大量自噬小体;各模型组细胞LC3-Ⅱ蛋白的表达:与control组比较,DTT(50、200、500μmol/L)组细胞中自噬相关蛋白LC3-Ⅱ的表达均显著升高(P均0.05);与DTT 200μmol/L组比较, 500μmol/L组LC3-Ⅱ蛋白的表达显著升高(P0.05).金刚藤对ERS-自噬肝癌细胞模型存活率的影响:JGT、DTT+JGT组细胞存活率均显著低于control组(P均0.05); JGT组细胞存活率显著低于DTT+JGT组(P 0.05).结论 DTT可诱导肝癌SMMC-7721细胞ERS-自噬;金刚藤可抑制SMMC-7721细胞的存活率, DTT诱导的ERS-自噬可抵抗金刚藤抑制SMMC-7721细胞的存活率.     

来那度胺对人肝癌SMMC-7721细胞生长的影响

目的探讨来那度胺对人肝癌SMMC-7721细胞增殖的抑制作用及其可能机制。方法利用MTS法检测来那度胺对SMMC-7721细胞增殖的影响;实时定量PCR、Western印迹检测来那度胺作用于SMMC-7721细胞后血管内皮生长因子(VEGF)的表达。结果 50μmol/L的来那度胺对肝癌细胞SMMC-7721增殖有抑制作用;实时定量PCR和Western印迹结果显示,以25、50、100μmol/L来那度胺处理SMMC-7721细胞12 h后,VEGF mRNA表达水平显著减低(P<0.05);Western印迹结果表明,25、50、100μmol/L来那度胺处理SMMC-7721细胞24 h后,VEGF蛋白表达水平明显下调(P<0.05)。结论来那度胺可能通过抑制肿瘤细胞增殖、抑制肿瘤血管生成发挥双重抗肿瘤作用。     

三氧化二砷对人肝癌SMMC-7721细胞增殖、迁移及侵袭能力的影响

目的 探讨三氧化二砷(As2O3)对人肝癌SMMC-7721细胞增殖、迁移和侵袭能力的影响及其分子机制.方法 利用MTS/PMS法检测As2O3对SMMC-7721细胞增殖的影响;划痕愈合和侵袭实验检测As2O3对SMMC-7721细胞迁移与侵袭能力的影响;实时定量PCR、Western印迹检测As2O3作用SMMC-7721细胞后MMP-9的表达.结果 2.0 μmol/L的As2O3对肿瘤细胞增殖有抑制作用;划痕实验结果显示,2.0μmol/LAs2O3处理SMMC-7721细胞12和24 h后,较未处理组划痕距离明显增加(P＜0.05),表明As2O3可抑制SMMC-7721细胞迁移.Transwell实验结果显示,2.0μmol/LAs2O3处理SMMC-7721细胞12h和24 h后,穿膜细胞数较未处理组明显减少(P＜0.05),表明As2O3可抑制SMMC-7721细胞侵袭能力.实时定量PCR和Western印迹结果显示,以2.0μmol/L As2O3处理SMMC-7721细胞后,MMP-9 mRNA和蛋白表达均明显下调(P＜0.05).结论 As2O3可通过抑制肿瘤细胞增殖、迁移及侵袭,抑制肿瘤的转移.     

3种选择性环氧合酶-2抑制剂对肝细胞癌生长的影响

背景肝细胞癌(HCC)中有环氧合酶(COX)-2表达,非甾体抗炎药阿司匹林可能通过抑制COX-2的表达而抑制HCC生长.目的比较3种选择性COX-2抑制剂美洛昔康、赛来昔布和罗非昔布对人肝癌细胞株SMMC-7721 生长的影响,观察高选择性COX-2抑制剂罗非昔布对裸鼠HCC原位移植瘤生长的影响.方法采用3H-胸腺嘧啶核苷(3H-TdR)掺入检测SMMC-7721细胞的DNA合成情况;采用免疫细胞化学染色检测增殖细胞核抗原(PCNA)的表达;采用DNA原位末端标记(TUNEL)染色检测细胞凋亡.给予HCC原位移植瘤裸鼠罗非昔布每日30 mg/kg 8周,测量肿瘤体积和重量.结果美洛昔康、赛来昔布和罗非昔布均能显著抑制SMMC-7721细胞的3H-TdR掺入,其抑制作用呈剂量依赖性,50%抑制浓度(IC50)分别为8.55×10-8mol/L、1.22×10-8mol/L和6.27×10-9 mol/L.3种选择性OX-2抑制剂(1×10-5 mol/L)作用24 h均可明显降低SMMC-772I细胞的PCNA表达,使细胞凋亡指数较对照组显著增高(14.6%±2.8%、21.6%±3.6%和27.1%±3.5%对1.0%±0.7%,P＜0.01),COX-2抑制剂的选择性越高,凋亡指数也越高(P＜0.01).罗非昔布组裸鼠的HCC原位移植瘤显著小于对照组,体积抑瘤率和重量抑瘤率分别为73.2%和78.1%.结论3种选择性COX-2抑制剂均能在体外有效抑制SMMC-7721细胞的生长,选择性越高,抑制作用越强.罗非昔布在体内能抑制HCC的生长,可能成为治疗HCC的有效药物.     

熊果酸诱导人肝癌SMMC-7721细胞凋亡的研究

目的：观察熊果酸（UA）对人肝癌SMMC-7721细胞的增殖抑制及诱导其凋亡作用。方法：MTT法检测5、10、20、30、40、50μmol/L UA对SMMC-7721细胞生长的抑制作用,吖啶橙（AO）荧光染色、电镜和流式细胞仪检测细胞凋亡。结果：UA能显著抑制SMMC-7721细胞的增殖,其作用呈剂量依赖性。35.2μmol/L UA作用SMMC-7721细胞48小时后AO染色,荧光显微镜下可见细胞出现体积缩小,核碎裂,染色质凝集等凋亡形态改变;电镜下SMMC-7721细胞出现明显的细胞凋亡的形态学改变,细胞核染色质出现边聚和中聚,细胞内部分线粒体肿胀;SMMC-7721细胞凋亡率为（67.91±5.24）%,与对照组（2.95±0.56）%比较差异有显著性意义（P〈0.05）。结论：UA通过诱导SMMC-7721细胞凋亡抑制其生长。     

人参皂苷Rg3对人肝癌细胞Pim-3及Bad凋亡蛋白表达的影响

目的:研究人参皂苷Rg3对人肝癌细胞SMMC-7721中Pim-3及磷酸化Bad蛋白pBad(Ser112),pBad(Ser136)表达的影响.方法:用浓度为0、5、10、20、40和80 μmol/L的人参皂苷Rg3处理SMMC-7721细胞24 h后.采用四甲基偶氮唑盐(MTT)方法检测人参皂苷Rg3对SMMC-7721细胞增殖的抑制作用,倒置显微镜和流式细胞术观察人参皂苷Rg3对SMMC-7721细胞凋亡的诱导作用,Western blot方法检测不同浓度人参皂苷Rg3处理后SMMC-7721细胞中Pim-3,pBad(Ser112)和pBad(Ser136)的表达情况.结果:5、10、20、40和80μmol/L的人参皂苷Rg3对SMMC-7721细胞增殖的抑制率分别为4.69%、15.53%、22.17%、50.97%、61.65%.5.80 μmol/L的人参皂苷Rg3处理细胞呈现明显的凋亡形态学改变,80μmol/L人参皂苷Rg3处理SMMC-7721细胞24 h后,处理组比正常对照组凋亡明显增加,差异有统计意义(16.5%±4.3% VS 0.4%±1.3%,P<0.01).人参皂苷Rg3对细胞中Bad总蛋白的表达没有明显影响.Pim-3的表达随人参皂苷Rg3浓度的增加而逐渐减弱,而pBad(Ser112)的表达随人参皂苷Rg3浓度的增加而逐渐增强;pBad(Ser136)不表达,结论:人参皂苷Rg3的抗癌活性与其促进磷酸化Bad蛋白表达有关.     

三磷酸腺苷结合盒G2调控肝癌细胞多药耐药的作用

目的探讨三磷酸腺苷结合盒(ABC)G2在肝癌耐药细胞中的表达及其在肝癌细胞耐药中的作用。方法 MTT法检测阿霉素(ADM)作用肝癌SMMC-7721细胞24 h后半数生长抑制浓度(IC50)。利用药物浓度递增细胞培养方法建立肝癌耐药细胞,在SMMC-7721细胞培养液中加入ADM,逐渐提高ADM浓度,浓度从0.001μg/ml提高至0.100μg/ml,使细胞在0.100μg/ml ADM中稳定生长,命名为SMMC-7721/ADM细胞。倒置显微镜下观察SMMC-7721/ADM及其亲代SMMC-7721细胞形态。MTT法检测ADM作用肝癌耐药细胞SMMC-7721/ADM 24 h后的IC50,计算耐药指数。流式细胞术(FCM)检测SMMC-7721/ADM及其亲代SMMC-7721细胞凋亡、周期及细胞中ABCG2蛋白表达水平。结果 ADM对SMMC-7721细胞的IC50=(1.11±0.09)μg/ml。历时3个月时间成功使SMMC-7721/ADM细胞在含0.100μg/ml ADM的细胞培养液中稳定生长,细胞命名为SMMC-7721/ADM。倒置显微镜下观察,SMMC-7721/ADM细胞体积增大,细胞形态变得更不规则。SMMC-7721/ADM与SMMC-7721细胞相比细胞凋亡率无显著差异(P0.05),细胞G0/G1期显著降低(P0.05),细胞S及G2/M期无显著差异(P0.05)。SMMC-7721/ADM与SMMC-7721细胞相比,细胞中ABCG2蛋白表达水平显著增高(P0.05)。结论 ABCG2在肝癌多药耐药细胞中异常增高,高表达的ABCG2参与了肝癌多药耐药的形成。     

蛹虫草小分子肽增强胃癌细胞株SGC-7901对氟尿嘧啶化疗的敏感性

目的探讨蛹虫草小分子肽增强胃癌细胞株SGC-7901对氟尿嘧啶化疗的敏感性。方法测定SGC-7901对5-FU化疗敏感性及不同浓度蛹虫草小分子肽(5,10,20 mg/ml)对其的影响并观察细胞形态变化,计算IC50值。结果蛹虫草小分子肽对SGC-7901的抑制作用具有剂量依赖性,在实验所采用的剂量范围内,蛹虫草小分子肽的浓度越高,对肿瘤细胞增殖的抑制作用越明显(P0.05)。对照组SGC-7901对5-FU化疗的IC50为(1 206.3±53.2)μmol/L;实验组中加入5,10,20 mg/ml的蛹虫草小分子肽后,其IC50分别为(1 069.4±79.1)μmol/L,(987.8±40.5)μmol/L,(943.1±49.5)μmol/L;组间比较显示,各实验组的IC50值明显低于单用5-FU化疗组(P0.05),蛹虫草小分子肽浓度越高,IC50下降越明显(P0.05)。结论蛹虫草小分子肽对SGC-7901的增殖具有抑制作用,其浓度越高,对SGC-7901的抑制作用越明显。     

Control of intraabdominal hemorrhage and shock: a comparison of fluid resuscitation, MAST, and balloon occlusion

Our study examined the efficacy of four treatment modalities in controlling hemorrhage and achieving hemodynamic stabilization in hemorrhagic shock: intravenous fluid replacement (IV); military antishock trousers used concomitantly with fluids (MAST); balloon occlusion at the level of the diaphragm with concomitant fluid replacement (balloon); and a combination of MAST inflation, balloon occlusion, and fluid resuscitation (MAST and balloon). Twenty-eight mongrel dogs were anesthetized, and the spleen was exposed and completely crushed. The abdomen was closed, and treatment was initiated and continued for four hours or until the dog died. For all conditions the hematocrit dropped during the course of the experiment; balloon occlusion was effective at slowing this drop (P less than .0001), but MAST had no statistically significant effect. Animals with balloons bled more slowly into the abdominal cavity than did animals in the other two groups (P less than .0001). MAST also were effective at slowing the bleeding (P less than .05). Of the balloon and the MAST and balloon dogs, all except one survived the entire four hours; this difference between balloon and nonballoon dogs is significant (P = .002). MAST did not have a statistically significant effect on survival. Perfusion pressure (PP) declined during the course of the experiment, and the balloon was effective at slowing this decline (P less than .0001); none of the other comparisons was statistically significant.     

Use of fab fragments of digoxin-specific antibodies in the therapy of massive digoxin poisoning

A case of massive digoxin ingestion with multiple arrhythmias, consisting of high grade A-V block and ventricular ectopy not responsive to lidocaine, is described. The arrhythmias ceased following administration of digoxin-specific Fab fragments. The patient improved and was transferred to the psychiatric unit.     

Comparison of intravenous and oral routes of theophylline loading in acute asthma

In a prospective, randomized clinical trial, 19 patients with an acute exacerbation of asthma were given a loading dose of aminophylline by the IV (n = 10) or oral route (n = 9) of administration following treatment with epinephrine. Plasma concentrations of theophylline were measured prior to giving the loading dose, and one, two, three, and 24 to 48 hours later. Therapeutic effectiveness was evaluated by analyzing spirometric measurements prior to giving the loading dose, and one, three, and 24 to 48 hours later. Side effects also were recorded. In the IV group, the mean peak plasma theophylline concentration was 15.1 micrograms/mL one hour after loading, and in the oral group the mean peak serum theophylline concentration was 14.2 micrograms/mL three hours after loading. There was no correlation between theophylline concentrations and normalized change in spirometric values. There was no significant difference in spirometric values between the IV and oral groups. Nausea was slightly more common in the IV group. We conclude that there is no therapeutic advantage to giving a loading dose of aminophylline by the IV route rather than orally in patients with mild-to-moderate exacerbation of asthma initially treated with epinephrine.     

Role of microRNA in regulation of myeloma-related angiogenesis and survival

Multiple myeloma (MM) is a malignant disease caused by clonal proliferation of plasma cells that result in monoclonal gammopathy and severe end organ damage. Despite the uniform clinical signs, the disease is very diverse in terms of the nature and sequence of the underlying molecular events. Multiple cellular processes are involved in helping the malignant cells to remain viable and maintain proliferative properties in the hypoxic microenvironment of the bone marrow. Specifically, the process of angiogenesis, triggered by the interactions between the malignant MM cells and the stroma cells around them, was found to be critical for MM progression. In this review we highlight the current understanding about the epigenetic regulation of the proliferation and apoptosis of MM cells and its dependency on angiogenesis in the bone marrow that is carried out by different microRNAs.     

安徽巢湖市丝虫病控制、消除历程和监测报告

【摘要】 目的  回顾并总结巢湖市丝虫病的流行、控制,以及消除丝虫病的历程。 方法  统计分析1970~1989年在该市采取以消灭传染源、切断传播途径为主的防治策略,以及展开大规模防治的各个阶段。 结果  1989年,采用分层整群抽样调查方法调查,微丝蚴率下降至0.032%,以行政村为单位,微丝蚴率已控制在1.0%以下,达到卫生部规定的基本消灭丝虫病标准;1991年,经省级调查考核,认定全市已达到卫生部规定的基本消灭丝虫病标准。1996年,省级审评确认已达到卫生部颁布的消灭丝虫病标准。经过10年的监测和跟踪治疗,到2005年,全市丝虫病病原学监测未查见微丝蚴阳性者,解剖镜检人房内淡色库蚊1 348只,蚊体内均未查见人体幼丝虫。 结论  巢湖市达到了消除丝虫病的标准。     

RNA-Seq在老年神经退行性疾病研究中的应用

阿尔茨海默病及帕金森病是老年人最常见的两种神经退行性疾病,但其发病机制及治疗是研究的热点。随着高通量测序技术的进步及成本的下降,RNA-Seq也成为神经退行性疾病机制研究及生物标志物发现的有力手段。RNA-Seq相对于microarray具有高灵敏度、高准确性、高重复性以及噪声低等优势,在阿尔茨海默病及帕金森病研究中有较为广泛的应用,包括检测差异表达基因,可变剪接、新长链非编码RNA预测分析和miRNAs调控等,但是容易受病理复杂性及样本等因素影响。目前阿尔茨海默病及帕金森病转录组研究相比于癌症等还不够深入,在临床诊断及治疗应用还面临较大挑战。但是随着新技术及新方法的发展,RNA-Seq将进一步推动神经退行性相关疾病的研究和临床转化。     

岳阳市城区小学生血吸虫病防治健康教育现状调查

目的 调查湖南省岳阳市城区小学生血吸虫病防治知识、行为现状及健康教育需求情况,为制定科学有效的小学生血吸虫病健康教育方案提供参考依据。方法 采用分层整群抽样方法,抽取岳阳市城区洞庭湖湖畔学校和中心城区学校各2所,每所再从五、六年级分别抽取2个班的学生,通过调查问卷了解小学生血吸虫病相关知识、预防行为及健康教育需求,并进行统计分析。结果 共调查湖畔小学353人、中心城区小学363人,两组学校小学生年龄、性别、年级构成差异无统计学意义（t=-0.494,χ²_性别=1.615,χ²_年级=2.152;P均>0.05）。学生血吸虫病防治知识总知晓率为42.60%（305/716）,其中,湖畔学校小学生血防知识知晓率（52.97%）高于中心城区（32.51%）,差异有统计学意义（χ²=30.661,P<0.05）;学生行为正确率为76.68%（549/716）,血防知识知晓组行为正确率（81.31%）高于不知晓组（71.24%）,差异有统计学意义（χ²=6.384,P<0.05）。学生血防知识主要来源于老师（47.49%）;91.90%的学生愿意了解更多的血防知识,最喜欢的血防知识学习方式是参加课外活动（50.42%）。结论 岳阳市城区小学生的血防知识知晓率及行为正确率偏低,对血吸虫病防治健康教育需求较高。建议针对学生开展形式多样的血防健康教育,以增强学生的血吸虫病防护意识,提高自我防护能力。     

Advanced Evaluation of the Freeze–Thaw Damage of Concrete Based on the Fracture Tests

This paper presents the results of an experimental program aimed at the assessment of the freeze–thaw (F–T) resistance of concrete based on the evaluation of fracture tests accompanied by acoustic emission measurements. Two concretes of similar mechanical characteristics were manufactured for the experiment. The main difference between the C1 and C2 concrete was in the total number of air voids and in the A300 parameter, where both parameters were higher for C1 by about 35% and 52%, respectively. The evaluation of the fracture characteristics was performed on the basis of experimentally recorded load–deflection and load–crack mouth opening displacement diagrams using two different approaches: linear fracture mechanics completed with the effective crack model and the double-K model. The results show that both approaches gave similar results, especially if the nonlinear behavior before the peak load was considered. According to the results, it can be stated that continuous AE measurement is beneficial for the assessment of the extent of concrete deterioration, and it suitably supplements the fracture test evaluation. A comparison of the results of fracture tests with the resonance method and splitting tensile strength test shows that all testing methods led to the same conclusion, i.e., the C1 concrete was more F–T-resistant than C2. However, the fracture test evaluation provided more detailed information about the internal structure deterioration due to the F–T exposure.     

欧洲儿童肝豆状核变性诊疗推荐意见简介

正肝豆状核变性(Wilson’s disease,WD)是常染色隐性遗传相关的铜代谢障碍性疾病,估计患病率为1:30000。该病是由于编码P型ATP酶的ATP7B基因突变,影响铜的分泌及排泄所致铜沉积。儿童肝豆状核变性的临床表现可以从无症状肝脏疾病到肝硬化或者肝衰竭,但是神经精神症状却非常少见。常规的检测方法包括血清铜蓝蛋白、24 h尿铜。最终的诊断需要基于症状、评估铜代谢的生化学检