摘要:目的:观察不同浓度地塞米松对体外正常和缺氧培养的人视网膜色素上皮细胞(hRPE)生长及血管内皮生长因子(VEGF)表达的影响.方法:体外正常(氧浓度95％)和缺氧(氧浓度2％～5％)下培养视网膜色素上皮细胞,分别加入不同浓度地塞米松,用四唑溴盐(MTT)和RT-PCR的方法,分别检测药物对细胞生长的影响和细胞VEGF的表达.结果:同一药物浓度下,缺氧刺激后,细胞较正常培养细胞呈现增殖状态,VEGF表达增强;1～200 μg/ml药物浓度可刺激细胞增殖,VEGF表达增强;200～400 μg/ml药物浓度逐渐出现细胞增殖率下降,VEGF表达降低趋势.各组与对照组相比差异性显著(P＜0.05).结论:对于体外正常培养或缺氧培养视网膜色素上皮细胞,地塞米松在低浓度可刺激细胞生长,高浓度可抑制细胞生长,这一过程可能与VEGF的表达有关.

摘要:目的:进一步明确金雀异黄素(Gen)抑制缺氧人视网膜色素上皮细胞(RPE)血管内皮生长因子(VEGF)表达的机制.方法:以体外正常情况为对照,建立人RPE缺氧模型并经不同浓度Gen干预后,通过RT-PCR检测VEGF及其转录调节因子c-fos、c-jun、缺氧诱导因子(HIF)-1α、HIF-2α、HIF-3α mRNA的表达水平,了解Gen是否通过对VEGF的转录因子活性调节而影响VEGF的表达.结果:缺氧能使人RPE细胞的VEGF、c-fos、c-jun及HIF-2α、HIF-3α mRNA水平增高,对HIF-1α无影响;Gen能抑制缺氧诱导的VEGF表达;在所检测的VEGF转录因子中,Gen只对HIF-2α的转录有抑制作用.结论:Gen抑制VEGF表达的作用部分是通过抑制其转录调节因子HIF-2α转录实现的.

摘要:目的:探讨玻璃体腔注射睫状神经营养因子(ciliary neuyotrophic factor,CNTF)对视网膜色素变性RD小鼠视网膜形态结构的保护作用.方法:将出生10天(P10)的RD小鼠随机分成3组:CNTF治疗组、伪治疗组和空白对照组,治疗组和伪治疗组的玻璃体腔分别注射鼠CNTF和磷酸缓冲液(PBS),分别在出生后14、18、22、30天过量麻醉处死小鼠,光镜测量RD小鼠视网膜外核层厚度并做TUNEL检测.结果:TUNEL检测,RD小鼠视网膜外核层有TUNEL染色阳性细胞分布.光镜下观察显示,出生后第14、18、22、30天RD小鼠CNTF治疗组视网膜外核层厚度显著高于其他两组,差异有显著性(P＜0.05).结论:CNTF玻璃体腔注射延缓了感受器细胞的凋亡,对视网膜色素变性RD小鼠有治疗作用.

摘要:目的:初步研究近红外光谱分析法在鉴别周围神经运动束和感觉束的应用,探讨建立一种快速鉴别周围神经束性质的方法.方法:取Beagle犬的脊神经前根和后根(前根为运动神经,后根为感觉神经),利用近红外漫透射方式进行快速采样,并以它们的近红外吸收图谱用光谱模式识别法中的聚类分析法进行定性分类鉴别.结果:脊神经的前根和后根其一二阶导数光谱较为相近,不能直观进行鉴别;经聚类分析可以将前根和后根归为两类,达到直观鉴别的目的.结论:近红外光谱分析技术有可能成为一种快速鉴别周围神经束性质的方法.

摘要:目的:观察胎鼠神经干细胞移植入海人酸毁损成年大鼠海马中的迁移和分化情况.方法:体外培养胎鼠海马源性神经干细胞.立体定位注射海人酸毁损大鼠海马CA1区锥体细胞.毁损1周后,将Hoechst 33342标记的神经干细胞移植毁损区,分别于术后1、2、4、8周取材,利用荧光技术和免疫组织化学方法,追踪移植的神经干细胞在毁损侧海马中的存活、迁移和分化情况.结果:移植的神经干细胞在海马锥体层呈链状迁移,并分化为MAP2阳性细胞和GFAP阳性细胞.结论:移植的神经干细胞在海马锥体层呈链状迁移,大部分分化为胶质细胞.

摘要:目的:研究碱性成纤维细胞因子(bFGF)和转化生长因子β1(TGF-β1)在单一和联合应用时,对体外培养关节软骨细胞增殖的影响,初探符合组织工程需要的合适浓度的bFGF与TGF-β1的组合.方法:第四代幼兔关节软骨细胞,以bFGF(1,5,10 ng/ml)或TGF-β1(0.1、0.5、1.0 ng/ml)及bFGF(10 ng/ml)+TGF-β1(1 ng/ml)的单独和联合刺激组为实验组,以常规培养组为对照组,通过CellTiter96(R)AQueous单溶液细胞增殖分析法测定细胞增殖效应.结果:单独应用时,10 ng/ml的bFGF和1 ng/ml的TGF-β1分别为各自最有效的促增殖浓度(P＜0.01);以此浓度联合刺激条件下的细胞增殖活性与对照组及单因子浓度组相比,均有显著升高(P＜0.05).结论:合适浓度的bFGF与TGF-β1联合使用,能有效促进体外培养软骨细胞的增殖,有助于组织工程种子细胞问题的解决.

摘要:目的:研究碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGF)、胰岛素样生长因子-Ⅰ(insuline-like growth factor-Ⅰ,IGF-Ⅰ)单独及联合应用对体外培养多次传代的兔关节软骨细胞Ⅱ型胶原表达的影响,探讨传代多次的软骨细胞作为软骨组织工程种子细胞的可行性.方法:第3、5、7代体外培养的兔关节软骨细胞,以常规培养组作为对照组,bFGF(5 ng/ml)或和IGF-Ⅰ(100 ng/ml)单独刺激及联合刺激作为实验组,免疫组化法测定Ⅱ型胶原的定性表达,并通过图像分析作半定量分析.结果:对照组细胞第5代以后无阳性表达.第3、5、7代细胞在bFGF或IGF-Ⅰ刺激下,Ⅱ型胶原表达强度显著高于对照组(P＜0.01);bFGF和IGF-Ⅰ联合作用下,Ⅱ型胶原表达强度均显著高于单独以bFGF或IGF-Ⅰ刺激组(P＜0.05).结论:bFGF、IGF-Ⅰ能促进多次传代软骨细胞Ⅱ型胶原的表达,且联合应用时促进作用更为明显.多次传代软骨细胞在生长因子刺激下,仍有再分化的能力,而有成为软骨组织工程种子细胞的可能.

摘要:目的:探讨幽门螺旋杆菌Hp感染胃部疾病患者Th1/Th2及Treg细胞表达量的变化及其相关性.方法:收集Hp感染无症状者,慢性浅表性胃炎、慢性萎缩性胃炎、肠的化生、不典型增生及胃癌患者的全血,利用胃黏膜尿素酶试验、ELISA和免疫印迹检测Hp的感染;采用流式细胞术检测Th1细胞、Th2细胞的表达水平,以及Treg细胞的表达.结果:在不同胃部疾病伴Hp感染者中,随着疾病的进展,Th1细胞的表达没有显著性变化,而Th2细胞的表达逐渐增加,Th1/Th2逐渐降低,而Treg细胞的表达逐渐增加,且Th1/Th2与Treg细胞的表达二者之间呈显著负相关.结论:在Hp感染者中,Th1/Th2及Treg细胞在调节Hp感染的免疫应答中起重要作用,且与Treg细胞的表达增多呈显著负相关,与癌症的发生有一定的相关性.

摘要:目的:筛选血管内皮生长因子(vascular endothelial growth factor,VEGF)高表达肝癌细胞株,为进一步研究VEGF在肝癌治疗中的作用提供理论依据,并探讨VEGF受体在肝癌细胞株的表达及意义.方法:ELISA法及Western blot法分别检测人肝癌细胞株培养上清及细胞内VEGF蛋白的表达,免疫细胞化学方法检测VEGF R在人肝癌细胞中的表达.结果:5株肝癌细胞株均见VEGF蛋白表达,其中SMMC-7721的VEGF表达量最高,VEGF特异性受体Flt-1在HepG2、HHCC、SMMC-7721、Bel-7402见阳性表达,KDR在HepG2、HHCC、Bel-7402、QGY-7701见阳性表达.结论:肝癌细胞株中VEGF表达量不尽一致,VEGF在肝癌发生发展中可能存在自分泌作用方式.

摘要:目的:探讨环氧化酶(COX)-2抑制剂塞来昔布(celecoxib)对肺癌A549细胞的增殖抑制、COX-2表达和化疗敏感性的影响.方法:用四甲基偶氮唑盐微量酶反应比色法(MTT法)检测塞来昔布单药及联合顺铂(DDP)对COX-2高表达的人肺腺癌细胞A549的增殖抑制,流式细胞术分析塞来昔布联合DDP对细胞周期及细胞凋亡的影响,Western blot法分析不同浓度塞来昔布干预后对A549细胞内COX-2蛋白表达的影响.结果:MTT法检测显示,塞来昔布(0～100 μmol/L)对肺腺癌A549细胞的抑制作用呈时间依赖性、剂量依赖性效应,联合用药组与单药组相比差异有显著性(P＜0.001).流式细胞术分析显示,塞来昔布12.5 μmol/L与25 μmol/L联合DDP0.5mg/L时,A549细胞明显阻滞于G0-G1期,S期细胞比例减少,联合用药组的凋亡率明显高于单药组(P＜0.001).Western blot法分析显示,塞来昔布浓度超过50 μmol/L时,COX-2蛋白的表达受到了抑制.结论:塞来昔布的抗肿瘤作用机制涉及到COX-2依赖性途径.塞来昔布能增加DDP的化疗敏感性,塞来昔布与DDP联用对人肺腺癌A549细胞有明显协同抗肿瘤效应,该作用可能是通过增强对A549细胞的增殖抑制、诱导凋亡来实现的.

摘要:目的:确认新等位基因HLA-B*0740、DRB1*1449;DRB1*1609的血清学表型.方法:应用标准NIH微量淋巴细胞毒试验进行新等位基因血清学分型.结果:新等位基因HLA-B*0740、DRB1*1449、DRB1*1609的核苷酸序列与它们相应的同源等位基因不同,造成相应编码氨基酸残基差异,但是它们的血清学特异性仍为B7、DR14和DR16.结论:上述HLA新等位基因具有免疫学表型.

摘要:目的:观察咪喹莫特对小鼠哮喘模型气道炎症和肺组织Th2型胸腺和活化调节趋化因子(thymus and activation regulated chemokine,TARC)和其受体CCR4表达的影响.方法:建立哮喘模型,自第14天雾化吸入OVA前0.5 h,干预组分别雾化吸入咪喹莫特30 min及腹腔注射地塞米松.OVA雾化结束后24 h,每组小鼠眼球摘除采血收集血清,采用酶联免疫吸附法测定血清中TARC水平;收集肺泡灌洗液进行细胞计数和分类;HE染色观察肺组织炎症改变;用免疫组化方法检测肺组织CCR4和TARC的表达;用逆转录-聚合酶链反应半定量检测肺组织IL-4、IFN-γ、CCR4和TARC mRNA表达水平.结果:①哮喘小鼠HE染色可见气道上皮不完整,黏膜下有大量的炎细胞浸润,以淋巴细胞为主.地塞米松组和咪喹莫特组炎症程度减轻.②哮喘组小鼠支气管肺泡灌洗液(BALF)中细胞总数及各种炎症细胞较正常组显著增加,咪喹莫特治疗组的嗜酸性粒细胞、单核细胞和淋巴细胞比哮喘组明显减少.③哮喘组小鼠血清TARC水平较正常组升高,咪喹莫特治疗组小鼠血TARC水平较哮喘组降低,与正常组比较差异无显著性.④CCR4和TARC均在气道上皮细胞表达,咪喹莫特和地塞米松治疗组能减少CCR4和TARC的表达.⑤哮喘组小鼠肺组织IL-4、CCR4和TARC mRNA表达较正常组增强,咪喹莫特和地塞米松治疗组小鼠肺组织IL-4、CCR4和TARC mRNA表达较哮喘组降低,比正常组表达增加.结论:雾化吸入咪喹莫特可在一定程度上减轻哮喘小鼠的气道炎症,抑制肺组织CCR4和TARC蛋白和mRNA的过度表达.

摘要:目的:探求肾素-血管紧张素系统(RAS)基因多态性与冠心病(CHD)患者行经皮冠状动脉腔内成形术(PTCA)支架置入术后再狭窄(ISR)的相关性.方法:随机选取351例行PTCA加支架置入术的上海籍汉族患者,分为再狭窄和未狭窄两组.抽提患者外周血基因组DNA,利用PCR-RFLP和PCR-APLP鉴定方法进行血管紧张素原(AGT)基因、血管紧张素Ⅰ转换酶(ACE)基因和血管紧张素Ⅱ受体1(AT1R)基因分型,结合病史、临床指标,探讨RAS系统基因型和ISR的关系.结果:PTCA支架置入后再狭窄患者AGT基因三种基因型频率分别为MM型10.7％,MT型57.1％,TT型32.1％;未狭窄患者MM型、MT型、TT型的频率分别为4.7％、56.3％、39.1％.AGT基因多态性与ISR无明显相关性(χ2=1.34,P＞0.05).PTCA支架置入后再狭窄患者ACE基因3种基因型频率分别为:DD型30％、DI型43.3％、Ⅱ型26.7％;未狭窄患者3种基因型频率分别为:DD型8.2％、DI型41.1％、Ⅱ型50.7％.卡方检验结果显示再狭窄组D等位基因频率(51.7％)明显高于未狭窄组D等位基因频率(28.8％).PTCA支架置入后再狭窄患者AT1R基因3种基因型的频率分别为:AA型56％、AC型38％、CC型6％;未狭窄患者AA型、AC型、CC型的频率分别为50％、40％和10％,经比较两组差别无统计学意义.结论:中国上海汉族冠心病人群中,RAS系统中ACE DD基因多态型可能是PTCA后ISR发生的遗传指标.D等位基因可能是ISR的预测因子.多支狭窄的患者较单支狭窄的患者更易患ISR.

摘要:目的:探讨依达拉奉(Edaravone)对新生大鼠缺氧缺血性脑损伤(HIBD)的保护作用.方法:7日龄新生SD大鼠122只随机分为缺氧缺血组(HI组)、大剂量治疗组、小剂量治疗组和假手术对照组,制成HIBD动物模型,在缺氧缺血(hypoxia-ischemia,HI)后即刻分别腹腔注射依达拉奉或生理盐水,连续注射5天.HI后24、48、72 h各时间点检测脑组织匀浆MDA和SOD含量;电镜观察海马超微结构变化;TUNEL法检测HI后72 h各组脑皮层神经细胞凋亡;Morris水迷宫实验评价35天学习和记忆能力变化.结果:HI后24、48、72 h各时间点,大剂量治疗组与HI组MDA和SOD含量相比差异有极其显著性(P＜0.01),小剂量治疗组与HI组相比差异无显著性(P＞0.05);TUNEL染色结果示HI后72 h各组缺血侧脑皮层凋亡细胞数,大剂量治疗组[(206.83±21.94)个/2500个细胞]较HI组[(317.67±24.27)个/2500个细胞]明显减少,差异有显著性(P＜0.01),而小剂量治疗组[(301.30±17.61)个/2500个细胞]较HI组也减少,但差异无显著性(P＞0.05).结论:缺氧缺血后腹腔注射大剂量依达拉奉对新生鼠HIBD有明显保护作用.

摘要:目的:邻苯二甲酸二丁酯(DBP)孕晚期染毒诱导子代雄鼠隐睾的发生和探讨隐睾睾丸、附睾病理组织学的改变.方法:孕鼠40只,随机分组,在怀孕14～18天期间,每天分别灌胃给予大豆油(A组)、DBP 100 mg/kg(B组)、500 mg/kg(C组)、800 mg/kg(D组),出生70天后观察雄仔鼠隐睾发生率及隐睾睾丸、附睾病理组织学改变.结果:C、D组雄仔鼠隐睾发生率为10.8％和63.5％;隐睾睾丸、附睾的脏器系数显著减轻;睾丸生精上皮萎缩,生精细胞层减少甚至消失,曲精小管体积减小;同时附睾管腔中精子缺如.电镜下隐睾睾丸中出现异常的支持细胞.结论:睾丸是DBP主要作用的靶器官;中、高剂量DBP孕晚期染毒可致雄仔鼠隐睾的发生、睾丸生精上皮的损害,从而影响其生育能力.

摘要:目的:探讨严重左室功能低下的冠心病患者进行非体外循环冠状动脉搭桥术(OPCAB)的可行性.方法:回顾性分析2000年2月至2004年10月进行的选择性OPCAB手术34例的临床资料、手术资料、手术结果,总结围手术期治疗体会.结果:全部病例均完成OPCAB手术,34例患者行旁路移植2～5支,平均(3.4±0.7)支,使用主动脉内球囊反搏18例,其中16例均在手术中安放,应用主动脉内气囊泵(IABP)后血流动力学指标明显改善.围手术期死亡1例(2.9％).有2例术后出现急性左心心力衰竭,安放主动脉内球囊反搏后保守治疗好转.结论:严重左室功能低下患者实施OPCAB手术有良好的可行性和近期效果.

摘要:目的:建立肾、带血管蒂肾上腺移植大鼠模型,探索肾、肾上腺联合移植对移植肾的免疫保护作用.方法:分别建立假手术、肾移植和肾、带血管蒂肾上腺联合移植模型,对各组在血皮质醇、肾功能、移植肾病理等方面进行比较.结果:肾、带血管蒂肾上腺联合移植组血皮质醇浓度明显高于肾移植组(P＜0.05),血尿素氮及肌酐明显低于肾移植组(P＜0.05),移植肾排斥反应明显减轻.结论:肾、肾上腺联合移植对移植肾可产生免疫保护作用,其机制可能是通过移植肾上腺分泌内源性皮质激素产生免疫抑制作用.

摘要:目的:测量股骨后髁角,为全膝置换手术正确确定股骨假体旋转对线提供解剖数据,以减少术后髌股关节的并发症.方法:选择70例正常人的膝关节作为研究对象,年龄21～40岁.应用多层螺旋CT扫描(层厚0.5 mm)获取股骨远端的精确的横断面图像,Autocad软件测量股骨后髁角.此外,测量9个因肢体恶性肿瘤而行髋关节离断或股骨中上段截肢的膝关节,男6例,女3例.结果:股骨后髁角,男性:(6.5±1.3)°,女性:(7.1±1.7)°,性别之间差异无显著性(P＞0.05).结论:国人的股骨后髁角较西方人大,术中如以股骨后髁轴外旋3°来放置股骨假体,容易出现假体内旋.

摘要:目的:比较中老年与青壮年患者急性颈髓损伤的损伤类型的差异,为临床提供参考.方法:回顾性分析48例颈髓损伤患者,分为两组:A组(≥50岁)17例,B组(＜50岁)31例,均在伤后12 h内行MRI检查.结果:①A组无骨折无脱位型脊髓损伤7例(41.2％),B组4例(12.9％),两组差异有显著性(P＜0.05).②A组17例、(100％),B组8例(25.8％)伴有椎体骨质增生、椎间盘突出、黄韧带肥厚、后纵韧带钙化等改变,组间差异有显著性(P＜0.05).③A组:单纯颈髓肿胀8例,颈髓扭曲2例,颈髓受压6例,脊髓断裂1例.B组:单纯颈髓肿胀4例,颈髓扭曲2例,15例颈髓受压;7例骨片嵌入髓内,3例脊髓断裂.组间损伤类型差异有显著性(P＜0.05).结论:中老年颈髓损伤的类型与青壮年有显著不同:中老年无骨折无脱位颈髓损伤显著高于中青年组,颈髓损伤以较轻类型多见.MRI能有效显示急性期颈髓损伤的特点,为急诊治疗提供必要的依据.

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摘要:Objective: To explore the impact of V5-epitope tag inserted in the commercial pcDNA5/FRT/V5-His TOPO expression vector on the metabolic activation of AFB1 by human CYP2A13. Methods: A C-terminal 6×Histag was first introduced into CYP2A13 cDNA by PCR and subsequently transferred into the expressing vector pcDNA5/FRT. Another commercial pcDNA5/FRT/V5-His TOPO expression vector was used to develop the construct directly via PCR. Both of the constructs were then transfected into Flp-In CHO and allowed for the stable expression of CYP2A13. The mouse CYP2A5 and the vector alone were used as positive and negative control, respectively. The presence of CYP2A5 and CYP2A13 cDNA and their protein expression in the stable transfectant cells were determined by immunoblotting assay using a monoclonal antibody against 6×Histag. The AFB1-induced cytotoxicity in these tranfected CHO cells were conducted by MTS assay and the IC50 of cell viability was used to compare the CYP enzyme metabolic activity in AFB1 metabolism among these cells. Results: In accordance with the Flp-In system working mechanism, all the transfectant cells presented same protein expression level. The CHO cells expressing CYP2A5 was more sensitive to AFB1 treatment than those cells expressing CYP2A13, there was about 30-fold IC50 difference between the two cells (2.1 nmol/L vs 58 nmol/L). Interestingly, CYP2A13 fused with V5-Histag had the lost of metabolic activity to AFB1 than that fused with Histag alone, the IC50 of the viability in CHO-2A13-His-V5 cells was about 20-fold less than CHO-2A13-His (＞1 000 nmol/L vs 58 nmol/L). However, there was no change between CYP2A5 fused with V5-Histag and Histag alone (2.4 nmol/L vs 2.1 nmol/L). Conclusion: The results demonstrate that CYP2A13 fused with V5-epitope has a significant impact on its metabolic activation to AFB1, which indicated that it should be careful to select a new expressing vector for evaluating the enzyme activity in carcinogen metabolism.

摘要:Objective: To investigate the protective effect of CTLA4Ig in rejection of pancreaticoduodenal transplantation model of rat. Methods: Pancreaticoduodenal transplantion models were established from the donor F344 rats to the Lewis recipients. The models were divided into 2 groups: Group A and B with 12 rats in each group.2 days after transplantation, reciepients in group A were treated with i.p. injection of sailine, and those in group B CTLA4I were injected(200 μg). On day 1,4,7,10after transplantation, the grafts were harvested for histopathological examination. On day 4 after transplantation, the CD4+CD25+Tcells in the grafts were detected by Flow Cytometry. Results: Compared with group A: the degree of the rejection of grafts in group B was lower. The number of CD4+CD25+ T cells of graft was (7.91±1.26)﹪ in group A and (13.81±1.71)﹪ in group B, which had significant difference(P＜0.01). Conclusion: CTLA4Ig could inhibit T cell costimulatory pathway, prevent acute rejection, which might be mediated by increasing the number of CD4+CD25+ regulatory T cells.

摘要:Objective: To establish a rapid and precise continuous flow-injection chemiluminescence (CL) method to determine ribavirin. Methods: The ribavirin could restrain strongly CL reaction of luminol in sodium hydroxide (NaOH) solution with potassium chlorate(KClO3). The different experimental parameters affecting the CL intensity were studied carefully. Results:Under optimum conditions(The concentrations of luminol,KClO3, and NaOH were at 0.1 mmol/L,0.5 μmol/L,and 0.2 mol/L, respectively), the linear range of the working curves was 0.01-7.00 μg/ml with a detection limit of 0.004 μg/ml. Conclusion: The method is simple, rapid and sensitive, and successfully applied to the determination of ribavirin in pharmaceutical preparations and biological fluids.

摘要:Objective: To investigate V264M polymorphism of tissue factor pathway inhibitor (TFPI) in patients with acute coronary syndrome (ACS) in Chinese. Methods: The genotypes of V264M were detected by polymerase chain reaction (PCR) and restriction fragment length polymorphism (PCR-RFLP) in 136 patients with ACS and 106 healthy controls with their plasmal f-TFPI tested by enzyme linked immunosorbent assay (ELISA). Results: Two genotypes were found in V264M: GG and GA. The distribution frequencies of alleles and genotypes were in accordance with those predicted by Hardy Weinberg equilibrium in the present study (χ2 = 0.437, P＞0.05). Plasma f-TFPI level was lower in A allele bearer than that in non-A allele carriers and higher in ACS than control subjects (P＜0.05). No relationship was found between ACS and V264M polymorphism (P＞0.05).Conclusion: The V264M polymorphism may have an impact on the plasma level of free TFPI(f-TFPI), but it has no relationship with ACS.

摘要:Objective: To construct an eukaryotic expression vector that contains Smac gene, which is regulated by human Uroplakin Ib (UpIb) prommer. Methods: For the directionality of Smac expression in the transitional cell carcinoma of bladder, internal CMV and T7 promoter sequences in eukaryotic expression vector pcDNA3.1-Smac were replaced with UpIb promoter to consstuct a new plasmid. The plasmid DNA was identified by gel electrophoresis after being double digested at respective sites,and then the sequence was analyzed. The expression of Smac mRTA and protein in BIU87 cell line were detected after the transfection by using the newly constructed vector. Results: The Smac gene-carrying and UpIb promoter-regulated eukaryotic expression vector pcDNA3-UpIb-promoter-Smac was successfully constructed. The expression of Smac mRNA was approximately increased by 2.1 times and the expression of Smac protein was increased in about 71﹪ BIU87 cells. Conclusion: The new vector can be effectively expressed in bladder cancer cells and be of great significance for bladder cancer-targeted genc therapy.

摘要:Objective: To determine whether a mutation of mitochondrial DNA induces familial dilated cardiomyopathy in Chinese families with cardiomyopathy, and analyzed the correlation between the genotype and phenotype. Methods: Affected members in three Chinese families of the familial dilated cardiomyopathy underwent clinical evaluation and DNA analysis. Polymerase chain reaction and direct DNA sequencing were used to screen for mitochondrial DNA mutation. The type of mtDNA vairations and clinical situation were analysed on the patients with mitochondrial DNA mutation. Results: The mitochondrial A3434G mutation was identified in one of the three families,the 3434 th nucleotide A was replaced by G, which led to change of amino acid. No mutations were identified in the clinically unaffected members of the family and all members of the other two families.Conclusion: This study indicates that the mitochondrial A3434G mutation maybe related with familial dilated cardiomyopathy and deafness.

摘要:Objective: To investigate the frequency of mitochondrial 12S rRNA gene A1555G and 961 insC mutations among Chinese with sensorineural deafness. Methods: Blood samples from 78 sporadic cases with sensorineural deafness were obtained and DNA was extracted from the leukocytes, then the mitochondrial DNA target fragments were amplified by polymerase chain reaction(PCR). The 1555G mutations were detected by BsmA I restriction endonuclease digestion, every fragment was analyzed by sequencing; All the 961 insC mutation were detected by direct sequencing. Results: The percent age of A1555G mutation and mt961C insertion were 6.4﹪ and 2.6﹪ in the hearing-impaired Chinese subjects respectively. Conclusion: A1555G and 96linsC mutations in mitochondrial DNA 12S rRNA gene regions may play a role in the pathogenesis of hearing loss in the sporadic cases.

摘要:Objective: To explore the survival and migration of bone mesenchymal stem cells transplantated in intervertebral disc of rabbits and expression of the exogenic genes. Methods: Thirty-two rabbits were used, A randomized block design was used and discs in the same rabbit were one block,the lumbar discs from L2-3 to L5-6 were randomly divided into blank group, saline group, cell transplantation group Ⅰ and cell transplantation group Ⅱ. The fluorescence microscopy was used to determine the fluorescence of the maker protein GFP and DNA-PCR was used to analyze the copies of DNA of neomycin-resistant gene at 1, 3, 6, months after transplantation. Results: There was fluorescence in cell transplantation group Ⅰ and Ⅱ and none in blank group, saline group at 1, 3, 6 months after transplantation. In cell transplantation groups,the fluorescent distribution was more scatter with time, but no significant difference between cell groups Ⅰ and Ⅱ. The test of neomycin resistant gene expressed in cell transplantation group Ⅰ and Ⅱ and quantitative analysis showed that there was no significant difference between the cell groups Ⅰ and Ⅱ (P＞0.05). Conclusion: The transplanted bone mesenchymal stem cells can survive, migrate and the transfer genes can express efficiently, it suggests that the BMSC therapy may be effective to prevent and treat intervertebral disc degeneration.

摘要:Objective: To investigate the influence of Danshen Injection on airway inflammation and CD4+CD25+ regulatory T cells(CD4+CD25+ Tr) of asthmatic rats, and elucidate the possible mechanism of Dansben Injection in treatment of asthma.Methods: 30 Wister rats were randomly divided into control group, asthma group and Danshen Injection treated group. Bronchoalveolar lavage fluids (BALF) were collected, and cytology studies were conducted. Lung tissues were obtained and pathologic analyses were done with hematoxylin and eosin stain (HE). Flow cytometry was used to detect the CD4+CD25+ Tr ratio in peripheral blood mononuclear cells (PBMCs). Results: Total cell, the percentage of lymphocytes, neutrophils and eosinophils (Eos) in BALF of Danshen Injection-treated group were lower than that in asthma group (P＜0.05, P＜0.01). Compared with asthma group, less infiltration of inflammatory cells in lung tissues was observed in Danshen Injection-treated group. CD4+CD25+ Tr of asthma group was lower than that of control and Danshen Injection treated group (P＜0.05). Conclusion: Danshen Injection can suppress airway inflammation of asthmatic rats, probably by increasing the number of CD4+CD25+ Tr.

摘要:Objective: To evaluate the diagnostic and prognostic values in patients with delayed encephalopathy after acute carbon monoxide poisoning. Methods: The tibial nerve somatosensory evoked potentials (SEPs), vision evoked potentials (VEPs), and brain stem audition evoked potentials(BAEPs) were performed in 32 healthy adults and 43 patients with delayed encephalopathy after acute carbon monoxide poisoning. Results: This paper indicated abnormalities of tibial nerve SEPs in 31 patients (31/43, 72.1﹪), VEPs in 17 patients (17/28, 60.7﹪), and BAEPs in 14 patients (14/43, 32.6﹪). These results showed that the greatest diagnostic value was SEPs, followed by VEPs and, BAEPs with the lowest sensitivity. Conclusion: Multimodality evoked potentials (EPs) can be used for evaluating the diagnostic and prognostic values in cases of delayed encephalopathy after acute carbon monoxide poisoning.

摘要:Objective: Hyperthermia is an attractive addition to multidisciplinary approaches to clinical cancer treatment.The efficiency of hyperthermia depends on the elevation of the temperature and the duration of treatment. It has been reported that in vitro and in vivo hyperthermia enhanced the cytotoxic effect of certain anticancer drugs. However, this enhancement varies,depending on the drug used and the scheduling of treatments. Thus, the combination effect of chemotherapy and hyperthermia remains unclear. In this study, we aimed to investigate whether concurrent exposure of human hepatocellular carcinoma cells SMMC-7721 to chemotherapeutic agents andhypenhermia could increase anticancer effects. Methods: Two chemotherapeutic agents, cisplatin and hydroxycamptothecin, were applied. The MTT assay was performed to evaluate the growth inhibition of SMMC-7721 induced by anticancer drugs with and without hyperthermia. Flow cytometric analysis was used for the assessment of apoptosis after treatments. Results: The percentages of growth inhibition of SMMC-7721 induced by cisplatin (10 μg/ml) alone,hydroxycamptothecin (1 μg/ml) alone, hyperthermia alone, cisplatin and hyperthermia, hydroxycamptothecin and hyperthermia,were 20.77﹪, 13.65﹪, 32.46﹪, 62.76﹪, 71.89﹪, respectively. The percentages of apoptosis of five treatments are 5.56﹪,3.96﹪, 10.16﹪, 24.32﹪, 20.42﹪, respectively. Conclusion: While both hyperthermia and anticancer drugs can individually induce apoptosis and anti-proliferation effect, the combination of the two treatments induce significantly higher apoptosis and cytotoxicity than hyperthermia or anticancer drugs treatment alone. These data suggest a synergistic benefit when hyperthermia and anticancer drugs used concurrently.

摘要:Objective: To explore the loss of heterozygosity(LOH) on chromosome 6q in ovarian cancer, and localize a minimum area in deletion region. Methods: 93 ovarian tumors were analyzed for LOH studies with 10 microsatellite markers spanning chromosome 6q. To further localize a minimum area in deletion region. Nineteen microsatellite markers were used to refined a minimum area. Results: Forty three tumors (46﹪) were demonstrated allelic losses, which spanned less than two megabase areas, franked by a distal marker D6S311 and a proximal marker D6S1649, and likely harbored ovarian tumor suppressor gene (s). With analysis of density of LOH, increased DNA copy number at loci of 6q was demonstrated between D6S1649 and D6S311. Conclusion: It is possible that duplication after the allelic loss might be a main mechanism that leads to carcinogenesis in ovarian tumor. The refinement of these candidate tumor suppressor genes loci might facilitate future loss of heterozygosity studies and enable the isolation of candidate genes from this region.

摘要:Objective: To evaluate the results of repairing large defect on abdominal wall with artificial meshes (expansible polytetrafluoroethylene, e-PTFE and Composix Mesh). Methods: Four cases with large defect of abdominal wall caused by abdominal wall tumors or injuries were repaired with artificial meshes and myocutaneous flaps. Results: The cases were followed up 7 months to 2.5 years with no complications such as incisional hernia, bowel adhesion and exposure of the prosthetic materials.Conclusion: The combined use of artificial meshes contained e-PTFE and myocutaneous flaps is a reliable and effective method for repairing large defect on abdominal wall.

摘要:Objective: To investigate the relationship between different sedation depth and the restlessness after using midazolam as an adjuvant during regional anesthesia. Methods: One hundred and fifty patients undergoing regional anesthesia were randomly divided into three groups. In group A, the patients were sedated at the level of OAA/S Ⅳ with midazolam during regional anesthesia. In group B and C, the sedation depths were kept at the levels of Ⅲ and Ⅱ respectively. The changes of BP, HR and SpO2 were recorded before and after midazolam.The rate of restlessness was also evaluated. Results: Blood pressures decreased in certain degree at 5 and 10 min after midazolam compared with those before in all three groups. BP decreased over 20﹪ of the baseline in 4 cases in group A, in 5 cases in group B, and in 5 cases in group C. There was no significant difference in BP dropping among three groups. Compared with group B and C SpO2 dropped significantly at 3 and 5 min after midazolam in group C, in which SpO2 was less than 93﹪ in 8 cases. The restlessness rate in group C was 22﹪, which was significantly higher than those in group A (2﹪ ,P＜0.01) and group B(4﹪ ,P＜0.05). Conclusion: The sedation depth is related to the rate of restlessness when midazolam is used as an adjuvant during regional anesthesia. It is suggested that the appropriate sedation depth for the patients under regional anesthesia is OAA/S Ⅲ.

摘要:As common acute abdomen, most of acute pancreatitis(AP) are self-restricted. Only a few patients may develop into worse state with local complications or organ failures, and finally neastic acute pancreatitis (NAP). With the change of people's dietaries, cholelithiasis morbidity and popularization of wine in recent years, the number of AP patients has increased.Although people conducted enormous studies on pathogenesis of AP and brought forward many be valuable theories, yet the exact mechanism is still unclear by far. There are many therapies of AP which should be unexceptionally classified as operative therapy and non-operative therapy. With the increasing understanding of the disease in recent years, we found many defects of operation and good therapeutic effects of traditional Chinese medicine in AP. Traditional Chinese medicine as an auxiliary therapy has been generally paid close attention in clinical practices. Traditional Chinese medicine is a treasure-house of China. This article summarizes the main mechanisms of AP treatment by traditional Chinese medicine and the progress of laboratory studies. It aims to help people recognize the multiple-target treatment effects and conspicuous efficacy of traditional Chinese medicine and promote the popularization of traditional Chinese medicine in AP treatment.