• 2005年第5期文章目次
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    • >南京医科大学学报(自然科学版)
    • 色素上皮衍生因子与年龄相关性黄斑变性的研究进展

      2005, 25(5). DOI: 10.7655

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      摘要:

    • 转化生长因子-β1对人眼眶成纤维细胞增殖的影响

      2005, 25(5). DOI: 10.7655

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      摘要:目的:观察转化生长因子β1(TGF-β1)对人眼眶成纤维细胞(orbital fibroblasts,OFs)增殖的影响,探讨增殖性玻璃体视网膜病变(proliferxtive vitreoretinopathy,PVR)的发病机制.方法:OFs体外培养,用不同浓度TGF-β1(0.1、1.0、5.0、10.0、100.0 μg/L)对细胞进行处理,计算细胞的数量,绘制细胞生长曲线;椎虫蓝染色计算活细胞比例、MTT比色法测定吸光度(A)值;流式细胞仪(FCM)检测细胞周期分布.结果:0.1、1.0、5.0、10.0μg/L的TGF-β1作用后OFs生长曲线上移,活细胞数增加,A值上升,处于增殖期(S+G2)的细胞比例增多.100.0μg/L的TGF-β1对OFs作用相反.结论:TGF-β1对人眼眶成纤维细胞增殖有双相调节性,其不同作用的发挥依赖TGF-β1的浓度,TGF-β1的促成纤维细胞的增殖作用可能诱发PVR.

    • 糖尿病患者22例眼的电生理变化研究

      2005, 25(5). DOI: 10.7655

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      摘要:目的:观察糖尿病患者图形视觉诱发电位(PVEP)、图形视网膜电图(PERG)的改变.方法:对10例(20只眼)正常人及22例(37只眼)糖尿病患者(无糖尿病视网膜病变8眼,单纯型糖尿病视网膜病变21眼,增殖型糖尿病视网膜病变8眼)进行PVEP、PERG检测.结果:糖尿病患者与正常对照组PVEP之间的差异有显著性(P<0.001),表现为P100波潜伏期延长和幅值降低;糖尿病患者与正常对照组、PDR组与其他各组PERG之间的差异均有显著性(P<0.001),表现为q波潜伏期延长和幅值降低,糖尿病组PVEP的P100波潜伏期延长和糖尿病患者空腹血糖呈正相关.结论:联合检测PVEP、PERG对糖尿病患者早期诊断和预后评估有一定的临床应用价值.PERG能比PVEP更好的反映糖尿病视网膜病变的损害程度.

    • 表面麻醉下糖尿病患者白内障超声乳化手术

      2005, 25(5). DOI: 10.7655

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      摘要:目的:观察表面麻醉下糖尿病患者行白内障超声乳化及人工晶体植入的临床效果.方法:临床确诊的糖尿病患者220例260眼,在表面麻醉下经透明角膜隧道切口,行白内障超声乳化及折叠式人工晶体植入手术.结果:所有患者均能耐受手术,未发生麻醉并发症.术后1天视力≥0.5者148眼,占56.92%;术后1周视力≥0.5者206眼,占79.23%.手术并发症主要有:角膜水肿、前房纤维素性渗出、后囊破裂.结论:表面麻醉下糖尿病患者白内障的超声乳化手术具有简化操作过程,并发症少,视力恢复快的特点,术后效果满意.

    • 新生兔与成兔角膜缘上皮移植于纤维蛋白胶载体的实验研究

      2005, 25(5). DOI: 10.7655

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      摘要:目的:利用组织工程技术体外重建角膜上皮组织,为重建角膜表面提供良好的移植材料和方法.方法:以等量凝血酶和纤维蛋白原作用生成的纤维蛋白胶为载体,分别种植新生兔和成兔角膜缘组织块,体外培养重建角膜上皮层,观察两组细胞的生长覆盖面积,t检验比较两组细胞的生长差异,并从生长特性、形态学特点及免疫细胞化学方面进行观察检测和比较.结果:两组兔角膜缘上皮细胞在纤维蛋白胶上黏附生长增殖,生长覆盖面积差异无统计学意义.体外培养7天左右即能较好地形成单层上皮细胞,2周左右形成的复层上皮细胞与纤维蛋白胶构成角膜上皮组织经检测与生理状态下的角膜上皮组织相近.结论:以纤维蛋白胶为载体、新生兔和成兔体外培养的角膜上皮组织片有些可作为眼表重建角膜上皮移植良好的来源之一.

    • 兔眼非穿透性滤过手术联合羊膜移植的实验研究

      2005, 25(5). DOI: 10.7655

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      摘要:目的:探讨两种不同术式的羊膜移植在兔眼非穿透性滤过手术中的抗瘢痕作用.方法:选取新西兰白兔15只,将30只眼随机分为3组,第1组行非穿性滤过手术联合巩膜瓣下羊膜移植,第2组行非穿透性滤过手术联合结膜瓣下羊膜移植,对照组行非穿透性滤过手术.术后观察术眼结膜及前房反应,30天时行眼球组织切片及减压腔处组织学检查,测定其胶原的平均吸光度(A)值.结果:术后30天对照组减压腔处的胶原相对含量A为(62.72±10.75).非穿透性手术巩膜瓣下羊膜移植组为(49.91±8.60).对照组与两实验组间差异显著(P<0.05),两实验组间差异不明显(P>0.05).结论:巩膜瓣下,结膜瓣下放置羊膜组织均能抑制兔眼非穿透性滤过手术区瘢痕形成,且对兔眼组织无毒副作用.

    • SITA-Fast与Full Threshold两种视野检查程序在开角型青光眼中的应用探讨

      2005, 25(5). DOI: 10.7655

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      摘要:目的:评价SITA-FAST视野阈值程序在开角型青光眼检查及视功能评价中的应用价值.方法:采用Humphery视野(POAG)计的SF程序与FT程序对正常人、早、中晚期开角型青光眼POAG患者及可疑患者进行检查,将两种检查时间、图形、视野指数(MD、PSD)及视野缺损点数进行比较和分析.结果:①SF程序检查时间仅为FT程序的1/3,P<0.01;②可疑POAG组中SF程序所得MD值、PSD值及早期POAG组中SF程序所得MD值均比FT程序低,P<0.01;③在正常组、POAG组中二种程序所得视野指数具有明显的正相关性,在可疑POAG组中不具有明显的相关性.结论:在青光眼筛查、POAG患者的视野检查中SF程序可取代FT程序,在可疑POAG患者的诊断中最好采用FT程序.

    • 保存人羊膜组织在兔非穿透性小梁切除术中应用的组织学观察

      2005, 25(5). DOI: 10.7655

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      摘要:目的:探讨羊膜移植应用在兔非穿透性小梁切除术中的抗瘢痕化作用.方法:选取新西兰白兔10只,双眼行非穿透性小梁切除术,每只兔随机选取一眼术中联合6 mm×6 mm羊膜移植于巩膜瓣下.结果:术后第7天时,实验组与对照组均可见弥散隆起的滤过泡;第14天时,实验组滤过泡存留有5只眼;对照组仅有1眼,差异有统计学意义.组织学检查显示,实验组滤过区纤维组织形成明显减少,术后14天仍有空腔存在:对照组滤过区见大量纤维瘢痕组织形成,术后14天滤过腔基本消失.结论:羊膜组织对免眼非穿透性滤过手术有抗瘢痕形成作用.

    • 三羟异黄酮对人乳腺癌细胞株MCF-7细胞增殖和细胞周期的影响

      2005, 25(5). DOI: 10.7655

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      摘要:目的:研究三羟异黄酮(genistein,Gen)对乳腺癌细胞株MCF-7细胞增殖和细胞周期的影响.方法:采用MTT比色法观察Gen对MCF-7细胞增殖的影响,流式细胞仪分析细胞周期分布,Western blot分别检测周期素B(cyclin B)、周期素E(cyclin E)蛋白表达情况.结果:Gen(≥10 μmol/L)对MCF-7细胞生长有显著抑制作用,细胞生长阻滞于G2/M期,cyclin B蛋白表达增加,且呈剂量-效应关系;但对cyclin E蛋白表达无影响.结论:Gen抑制MCF-7细胞增殖、诱导G2/M期阻滞与其导致cyclin B蛋白积累有关,与cyclin E蛋白表达无明显关系.

    • 维拉帕米对前体脂肪细胞3T3-L1分化及脂质积聚的影响

      2005, 25(5). DOI: 10.7655

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      摘要:目的:观察钙离子通道阻滞剂盐酸维拉帕米对3T3-Ll前体脂肪细胞分化及脂质积聚的影响.方法:体外培养3T3-Ll前体脂肪细胞,在常规诱导分化方案基础上加用维拉帕米(终浓度30μmol/L),采用油红O染色各分化时段脂肪细胞,计数含脂滴脂肪细胞数.结果:维拉帕米干预后,脂肪细胞分化速度低于正常对照,虽未影响脂滴出现的时间(第4天),但含脂滴脂肪细胞数均显著低于同期正常对照(P<0.001).结论:维拉帕米具有抑制脂肪细胞分化及脂质积聚的作用.

    • 淡色库蚊抗药性相关新基因--视蛋白基因的克隆与分析

      2005, 25(5). DOI: 10.7655

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      摘要:目的:获取淡色库蚊抗药性相关视蛋白基因(NYD-OP)全长cDNA序列.方法:根据抑制性差减杂交(SSH)结合cDNA芯片分离获得的淡色库蚊抗药性相关视蛋白基因的289 bp片段,设计上、下游引物,分别以cDNA文库和反转录二链产物为模板,采用快速扩增cDNA末端法(rapid amplification of cDNA end,RACE)扩增视蛋白基因5′、3′端,经对位拼接获得全长序列,并用相应的软件进行分析.结果:获得6个淡色库蚊视蛋白基因全长序列,开放阅读框分别为1116 bp(GeneBank/NCBI AY297441,AY297442)及1119 bp(GeneBank/NCBI AY297443,AY297444,AY299337,AY299338),以上序列经推导分别编码371及372个氨基酸.进行蛋白质序列分析,与烟草天蛾视蛋白的同源性是77%.结论:获得6个淡色库蚊抗药性相关视蛋白基因全长cDNA序列.

    • 乙型肝炎病毒核心抗原及Flt3配体双表达核酸疫苗的构建与体外表达

      2005, 25(5). DOI: 10.7655

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      摘要:目的:构建可同时表达乙型肝炎病毒核心抗原(HBcAg)和Flt3配体(FL)胞外段的双表达核酸疫苗.方法:将HBcAg和FL胞外段的基因用PCR方法扩增并分别引入相应的限制性内切酶酶切位点,然后克隆入真核双表达载体pIRES,获得含双基因片段的表达质粒;再取该表达质粒中的内部核糖体切入位点(IRES)序列和基因片段克隆入pJW4303载体中,获得相应的双表达核酸疫苗,经筛选鉴定后,以293T细胞瞬时表达检测两基因的表达水平.结果:构建成功以pIRES、pJW4303为载体的4种HBcAg和FL胞外段的双表达核酸疫苗,体外表达证实HBcAg和FL胞外段均能高效表达,基因位于IRES上游时表达水平较高.结论:用IRES元件实现了HBcAg和FL胞外段双表达核酸疫苗的构建,pJW4303为载体的核酸疫苗两基因表达水平优于pIRES为载体的核酸疫苗.

    • 小鼠结肠Cajal间质细胞超微结构及其与肠神经的关系

      2005, 25(5). DOI: 10.7655

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      摘要:目的:探讨小鼠结肠组织Cajal间质细胞超微结构及其与肠神经系统的关系.方法:用透射电镜观察健康成年雄性ICR小鼠结肠组织Cajal间质细胞的超微结构、分布及其与肠神经系统的关系.结果:小鼠结肠组织Cajal间质细胞主要结构特点:细胞呈纺锤形,有巨大的卵圆形核及向外伸展的长突起,胞质内有丰富的线粒体、大量光面内质网和粗面内质网;环肌层和纵肌层内Cajal间质细胞与肠神经元之间形成突触样连接,Cajal间质细胞突起包绕神经纤维.结论:Cajal间质细胞具有独特的超微结构,其与肠神经系统解剖与结构上的特殊关系可能是Cajal间质细胞调节多种胃肠功能的基础.

    • OK-432激活抗原负载的树突细胞对CIK细胞的作用

      2005, 25(5). DOI: 10.7655

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      摘要:目的:探讨OK-432对人类外周血单个核细胞(PBMC)来源的树突状细胞(DC)体外培养的影响,及由此产生的DC对CIK细胞抗瘤效果的影响.方法:从PBMC中分离出CD14+的DC前体细胞,经粒细胞-单核细胞集落刺激因子(GM-CSF)和白细胞介素4(IL-4)的作用,得到不成熟DC,再予以负载肿瘤抗原及经促成熟因子(MPF)OK-432或LPS的刺激后得到成熟DC.同时将PBMC在细胞因子诱导下培养成细胞因子诱导的杀伤细胞(CIK).通过对DC表面标志物的分析、经DC作用的CIK增殖活性、抗瘤效果的检测,研究了不同MPF对DC成熟和功能的影响.结果:OK-432可促进体外培养的DC高表达CD1a、CD80、CD83、HLA-DR,其作用强于LPS;经OK-432作用的负载抗原的DC可促进CIK增殖、加强CIK的抗瘤效率.结论:OK432可促进体外培养的DC成熟,并借此增强CIK的抗瘤作用.

    • 人子宫内膜基质细胞及腺上皮细胞的分离纯化和体外培养

      2005, 25(5). DOI: 10.7655

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      摘要:目的:建立人子宫正常内膜、内膜异位症在位及异位子宫内膜的基质及腺上皮细胞分离、培养的方法,为进一步研究子宫内膜异位症发生、发展的分子生物学机制提供一个理想的实验模型.方法:15例正常子宫内膜、15例在位子宫内膜及10例异位内膜经胶原酶消化、筛网过滤、差时贴壁等技术进行分离、纯化和体外培养,光镜观察,应用鼠抗人波形蛋白抗体、鼠抗人细胞角蛋白单克隆抗体免疫组化染色对间质及上皮细胞进行鉴定.结果:正常子宫内膜和子宫内膜异位症在位子宫内膜标本分离、培养均成功;5例异位内膜标本获得成功,基质细胞和腺上皮细胞纯度均可达95%以上,并均可传代.结论:采用酶解、筛网分离及贴壁能获得纯度较高的基质与腺上皮细胞.人子宫内膜细胞分离体外培养的成功,有助于研究子宫内膜异位症的发病机制,为临床实验提供重要的理论依据.

    • 人类系统性红斑狼疮血清对内皮(逆)穿越模型中树突状细胞表型成熟的影响

      2005, 25(5). DOI: 10.7655

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      摘要:目的:研究人类系统性红斑狼疮(systemic lupus erythematosus,SLE)血清对正常人单核细胞(monocyte,Mo)分化形成的树突状细胞(dendritic cell,DC)的表型成熟的影响.方法:分离正常人外周血单个核细胞(PBMC),加入内皮细胞穿越模型中,分别以正常人血清和SLE血清加入培养体系中,36 h后收集2次穿越内皮单层得到的,经形态学鉴定的DC细胞,流式细胞术测DC的CD80、CD86、HLA-DR的表达情况.结果:SLE血清诱导的DC其CD80的表达水平明显低于正常人血清刺激组,而二者均高表达CD86、HLA-DR.结论:DC的表型成熟和血清环境有一定关系.

    • 邻苯二甲酸二丁酯诱发大鼠尿道下裂模型的建立

      2005, 25(5). DOI: 10.7655

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      摘要:目的:尝试用邻苯二甲酸二丁酯(DBP)建立尿道下裂大鼠模型.方法:孕鼠30只,随机分成2组:①实验组(DBP)15只;②对照组(大豆油)15只.在大鼠怀孕12~20天期间,2组每天分别给予灌胃,实验组予DBP 500mg/(kg·d),对照组予大豆油,灌胃容积均为每只1 ml/d.待孕鼠分娩完毕,即对新生大鼠计数并在解剖显微镜下测量雄性新生鼠的肛门生殖器距离(AGD)和称体重.在雄性仔鼠出生后70天逐个检查尿道下裂的发生情况.结果:①DBP成功诱导出了雄性大鼠尿道下裂畸形,对照组大鼠的生殖器未见异常改变;②实验组新生大鼠的AGD缩短,但无统计学差异;实验组新生大鼠体重明显减轻,有显著统计学差异.结论:①成功建立了尿道下裂大鼠模型;②为人类小儿尿道下裂的病因学研究和防治提供科学的实验依据.

    • 脓毒症早期大鼠肝脏Toll样受体4基因表达及干预性研究

      2005, 25(5). DOI: 10.7655

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      摘要:目的:探讨严重腹腔感染所致脓毒症大鼠肝脏Toll样受体4(Toll like receptor4,TLR4)基因表达变化规律及对其调节机制进行初步探讨.方法:采用大鼠盲肠结扎穿孔(CLP)模型造成脓毒症.将96只大鼠随机分为正常对照组、假手术组、CLP组、地塞米松(DXM)干预组,于CLP后2、4、6、12、24 h处死动物,留取肝脏标本,以半定量逆转录多聚酶链反应技术及相关软件分析不同组TLR4 mRNA、TNF-αmRNA的表达,检测血清丙氨酸转移酶(ALT)、天冬氨酸转氨酶(AST)及肝脏组织病理,评估肝脏损伤情况.结果:CLP组2 h后肝脏TLR4 mRNA、TNF-αmRNA及血清ALT、AST比正常对照组明显增多(P<0.01);DXM干预组各时间点TLR4 mRNA、TNF-α mRNA的表达及血清ALT、AST较单纯CLP组显著降低(P<0.01),24 h病理切片显示肝损伤轻于CLP组.结论:严重腹腔感染可致大鼠肝脏TLR4 mRNA持续高表达,与肝脏的损伤密切相关;早期应用DXM可抑制TLR4 mRNA的表达,减轻肝脏损伤.DXM抑制腹腔感染所致的炎症反应可能与抑制TLR4的表达有关.

    • 重建肝动脉血供大鼠部分肝移植模型的建立及其意义

      2005, 25(5). DOI: 10.7655

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      摘要:目的:探讨重建肝动脉血供的大鼠部分肝移植模型的手术操作方法及其意义.方法:SD大鼠120只,采用重建肝动脉血供的大鼠部分肝移植60例(实验组),同期行不吻合肝动脉的经典"二袖套法"大鼠部分肝移植60例(对照组),分别于术后第1、2、4和7天处死后行肝功能、组织学检查,应用流式细胞仪检测移植肝细胞增殖活性.结果:两组肝功能ALT、TB第1天即开始明显增高,以后逐渐降低,第4天、第7天实验组ALT低于对照组,差异有显著性(P<0.05).第4天实验组TB低于对照组,差异有显著性(P<0.05),组织学检查术后实验组可见更多的二倍体和多倍体肝细胞(P<0.05),第2天和第4天实验组增殖指数分别为(32.81±3.45)%和(36.33±2.52)%,较对照组(24.08±4.66)%和(29.75±1.82)%明显增高,差异有显著性(P<0.01).结论:肝动脉重建可明显改善大鼠部分肝移植后移植肝的功能,促进移植肝的再生.

    • 乌斯他丁对肝移植术中血浆细胞因子和SOD、MDA的影响

      2005, 25(5). DOI: 10.7655

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      摘要:目的:研究背驮式肝移植手术中乌斯他丁(ulinastatin,UTI)对血浆细胞因子和丙二醛(MDA)、超氧化物歧化酶(SOD)的影响.方法:行背驮式原位肝移植(PBOLT)手术病人20例,随机分为对照组(C组,n=10),UTI持续泵注组(U组,n=10).于麻醉后开腹前、无肝前期120 min、无肝期30 min、新肝期5、60、120 min 6个时点分别采颈内静脉血比较两组血浆肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-1、IL-6、MDA和SOD水平.结果:与无肝前期相比,在无肝期和新肝期对照组血中上述指标浓度较术前显著升高(P<0.05),应用UTI组浓度水平无明显变化(P>0.05).结论:①背驮式肝移植手术特别是新肝期,体内发生了急性炎性反应;②乌斯他丁能减轻此类手术中的急性炎性反应.

    • 多发性骨髓瘤20例细胞遗传学研究

      2005, 25(5). DOI: 10.7655

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      摘要:目的:分析多发性骨髓瘤(MM)细胞遗传学变化及其与MM的分型、分期及其他预后参数的关系.方法:采用骨髓细胞24 h短期培养法及R显带技术进行染色体核型分析.结果:初治的20例MM患者中染色体核型异常者9例(45%),均有数目异常;6例有结构异常;7例发生于Ⅲ期,2例发生于Ⅱ期;5例IgG型,2例IgA型,2例轻链型;有异常核型的MM患者表现为高β2微球蛋白(β2-MG)、高C反应蛋白(CRP)、低血红蛋白(Hb)、低血小板(Plt)、骨髓瘤细胞计数超过30%,高血钙(Ca2+)、高乳酸脱氢酶(LDH).与无核型改变的MM患者比较差异有统计学意义.结论:近半数MM患者存在染色体异常,与部分预后参数密切相关.预后较差.

    • 基因GABRB3多态性与非综合征性唇腭裂的遗传易感性

      2005, 25(5). DOI: 10.7655

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      摘要:目的:探讨中国汉族人GABRB3基因的多态性对非综合征性唇腭裂(NSCL/P)的遗传易感性的作用.方法:对76例非综合征性唇腭裂患儿和60例正常对照儿童用PCR-SSCP方法分别测定GABRB3的等位基因片段,采用病例对照研究进行分析.结果:GABRB3等位基因共检测出9个,病例对照研究结果显示:所有等位基因频率在病例组和对照组中经X2检验,P值均大于0.05.结论:GABRB3基因在中国华东地区人群NSCL/P中不起主要作用,可能不是它的易感基因.

    • 南京市五年间乳母放弃纯母乳喂养原因分析

      2005, 25(5). DOI: 10.7655

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      摘要:目的:了解南京地区影响纯母乳喂养的原因,提出有效的干预措施.方法:1998年、2000年、2002年分别对4个月以内婴儿母亲进行放弃纯母乳喂养原因问卷调查,数据采用SPSS软件进行分析和显著性检验.结果:放弃纯母乳喂养的首要原因是自觉奶不足,0~28天组,3次放弃纯母乳喂养的原因顺位一致,0~4个月组,因母亲问题放弃纯母乳喂养的由1998年的第4位上升到2002年的第3位,差异有显著性(P<0.05).结论:南京市乳母放弃其婴儿纯母乳喂养的常见原因是自觉奶不足、婴儿生病、母亲问题.加大卫生宣教覆盖面,做好孕产妇及婴儿的保健工作,可提高该市婴儿纯母乳喂养率.

    • >南京医科大学学报(英文版)
    • Jak1/STAT3 pathway mediates the inhibition of lipoxin A4 on TNF-α-induced DNA synthesis of glomerular mesangial cells in rats

      2005, 19(5). DOI: 10.7655

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      摘要:Objective: To examine whether lipoxin A4 (LXA4) has an inhibitory effect on tumor necrosis factor-α(TNF-α)-induced DNA synthesis of glomerular mesangial cells of rat, and explore the molecular mechanisms of LXA4 action. Methods: Glomerular mesangial cells of rat were cultured and preincubated with LXA4 at different concentrations, and then treated with TNF-α( 10 ng/ml). DNA synthesis was assessed by the incorporation of [3H]-thymidine in mesangial cells. Expression of cyclin E protein was determined by Western blotting analysis. Activities of signal transducers and activators of transcription-3 (STAT3) were analyzed by electrophoretic mobility shift assay (EMSA). Results: TNF-α-stimulated DNA synthesis of mesangial cells, upregulation of cyclin E protein and STAT3 activities were inhibited by LXA4 in a dose-dependent manner. Conclusion: TNF-α-induced DNA synthesis of mesangial cells can be inhibited by TXA4probably through the mechanism of Jak1/STAT3 pathway-dependent signal transduction.

    • Intrathecal co-administration of ketamine and morphine preventing activation of astrocytes and decreasing releases of IL-1β and IL-6 from spinal cord in rats of morphine tolerance

      2005, 19(5). DOI: 10.7655

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      摘要:Objective: To investigate the effects of intrathecal administration of ketamine, a non-competitive N-methy-D-aspartate receptor antagonist, combined with morphine on the activation of astrocytes and releases of IL-1β and IL-6 from spinal cord in the rats of morphine tolerance. Methods: Twenty-seven Sprague-Dawley male rats were randomly divided into sham-operated, morphine tolerance, and morphine plus ketamine group. The subarachnoid catheterization of all the rats was prepared by the method of Jianping Yang.Morphine 20 μg in 10 μl was administrated intrathecally to induce spinal morphine tolerance once daily for 5 consecutive days. Morphine and ketamine 250 μg in 10 μl total volume was given in morphine plus ketamine group. Three groups all received intrathecal morphine 5 μg in 10 μl for morphine challenge test at 24 h after last administration of the morphine. After morphine challenge test, lumbar spinal tissues were taken for measurement of glial fibrillary acidic protein (GFAP) of astrocyte in lumbar spinal horn cord by immunohistochemistry and IL-1βand IL-6 of spinal cord by ELISA. Results: The decrease of %MPE induced by chronic intrathecal morphine was inhibited by ketamine and hyperalgesia and allodynia induced by morphine-withdrawl were alleviated. The average areas, the average absorbency (-A), the integral absorbency (A) of GFAP immuno-reactive cells in the dorsal horn, and IL-1β and IL-6 of spinal cord were significantly larger in morphine tolerance group than in morphine plus ketamine group. Conclusion: Co-administration of ketamine and morphine enhance antinociceptive effect of morphine and prevent the development of morphine tolerance. Ketamine might attenuate the activation of astrocytes and inhibit the release of IL-1β and IL-6 from spinal cord in repeated intrathecal morphine rats.

    • The antiapoptotic effect of insulin against anoxia/reoxygenation injury in cultured cardiomyocyte of neonatal rat

      2005, 19(5). DOI: 10.7655

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      摘要:Objective: To study protective effect of insulin against cardiomyocyte apoptosis in anoxia/reoxygenation (A/R)injury of neonatal rat. Methods: The model of A/R injury was finished through receiving anoxia for 2 h and reoxygenation for 4 h in cultured cardiomyocytes of neonatal rat. The cardiomyocytes were divided randomly into 3 groups: control group (CON), anoxia/reoxygenation group (A/R) and insulin-treated group (INS). At the end of reoxygenation of 4 hours, activities of lactate dehydrogenase (LDH),contents of malondialdehyde (MDA) were assessed through spectrophotometric procedures, myocyte apoptosis were detected through TUNEL and DNA Ladder. Results: MDA, LDH, and Apoptosis Index were significantly decreased in INS group compared with A/R group (P<0.01). Conclusion: Insulin has a protective effect against A/R injury in cultured cardiomyocyte of neonatal rat; the protective mechanism may contribute to antiapoptosis of insulin.

    • Adeno-associated virus mediated apoA-I and apoA-IMilano expression in skeletal muscular cells

      2005, 19(5). DOI: 10.7655

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      摘要:Objective: To construct recombinant adeno-associated virus type 2(rAAV2) vectors encoding human apoA-I/apoA-IMilano protein, and explore an effective and safe method to prevent and treat the atherosclerotic diseases. Methods: Human apoA-I cDNA with a His-tag in the upward stream of cDNA sequence were obtained with RT-PCR and PCR, and human apoA-IMilano cDNA was prepared by QuikChange[ ] Site-Directed Mutagenesis Kit. After extracted rAAV vectors with a most economic and convenient method, the particle numbers of rAAV vectors were measured by Dot-blot, and the purity was assayed by SDS-Page. The expression efficiency of the apoA-I or apoA-IMilano in C2C12 infected by rAAV vectors were detected by ELISA method. Results: ApoA-I cDNA was gained by RTPCR and a His-tag was added in the upward stream of apoA-I cDNA successfully. ApoA-I cDNA was mutanted to apoA-IMilano cDNA successfully by QuikChange[ ] Site-Directed Mutagenesis Kit. The both titres of the rAAV vectors of apoA-I and apoA-IMilano were about 2×1014/L, and the result of SDS-Page showed that the purity of the rAAV vectors was satisfied. The expression level of apoA-I was (0.39±0.04) μg/ml and the apoA-IMilano was (0.31 ±0.03) μg/ml in the DMEM culture medium at the first 24 h after transfection.Conclusion: The success of the rAAV vectors construction, purification and the expression of apoA-I and apoA-IMilano in C2C12 cells mediated by these vectors, makes possible to inject rAAVA and rAAVAM vectors into mice muscle, and rises a new hope on finding a new way to prevent and treat atherosclerotic diseases and cardiovascular disease.

    • The effect of hyperglycemia on blood brain barrier of rats with focal cerebral ischemia/reperfusion injury

      2005, 19(5). DOI: 10.7655

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      摘要:Objective: To determine whether hyperglycemia could aggravate the microvascular damage in ischemic stroke.Methods: Hyperglycemia model was made by injection of streptozocin through subcutaneous injection in wistar rats. Using the suture model, the rats were subjected to 3 h of focal ischemia and different times of reperfusion,including 6,12,24,48,96 h and 7 d. TTC dyeing was used to show the infarction area of rats. The infarctive volume of rats were calculated by computer imaging analysis system;Matrix metalloproteinase (MMP-2) and (MMP-9)were detected by immunohistochemistry and in situ hybridization histochemistry in Wistar rats.Results: The infarctive volume was siginificantly larger in hyperglycemic rats than that of nonhyperglycemic rats. The level of MMP-2,MMP-9 expression in the group of hyperglycemic rats was higher than that of nonhyperglycemic rats. Conclusion: Hyperglycemia aggravated the injury of focal ischmia-reperfusion in wistar rats and the higher expression of MMP-2, MMP-9 might be one of the mechanism in aggravation of focal ischemia/reperfusion injury.

    • Minocycline protects the apoptosis of PC12 cells induced by 1-methyl-4-phenylpyridinium

      2005, 19(5). DOI: 10.7655

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      摘要:Objective: To explore the protective effect of minocycline on the apoptosis of cellular parkinsonism models induced by MPP+ . Methods: Using PC12 cells as the apoptotic model of dopaminergic neurons, MC and MPP+ were added into the culture medium of PC12 cells, and using MTT to assay the cell viability and metabolic state; The cells apoptosis was assayed by electrophoresis method and using flow cytometry FACS to assay the apoptosis ratio. Results: Added the MPP+ to get the concentration of 10μmol/L, the cellular parkinsonism model of apoptosis had been prepared. The pre-treatment of MC (100 μmol/L) could significantly increase the PC12 cell viability. The apoptosis ratio of MC + MPP+ group was significantly lower than that of MPP+ group, but was still significantly higher than that of control group. Conclusion: MC may protect the cell apoptosis induced by MPP+ to some extent.

    • Release of myocardial noradrenaline during acute hibernation in the isolated rat heart

      2005, 19(5). DOI: 10.7655

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      摘要:Objective: To explore the release of myocardial noradrenaline during acute hibernation. Methods: The hearts were gained from rats and set up as modified Langendorf preparations beating isometrically. They were perfused with modified Krebs-Henseleit buffer under controlled pressure. Mechanical measurements and coronary effluent were recorded simultaneously at 30 min intervals for 150 min. Lactate dehydrogenase in coronary effluent was assayed at the beginning, 60 min and 120 min low-flow ischemia. Noradrenaline in coronary effluent was determined at the beginning of low-flow and 120 min of low-flow ischemia and also in control, during hibernation and after 30 min reperfusion during stimulation, myocardial noradrenaline response on tyramine was investigated in absence or presence of desipramine after 30 min reperfusion. Results: In the control, there was nosignificant chant in noradrenaline overflow during 120 min perfusion; In the acute myocardial hibernation group, there was also nosignificant difference in noradrenaline overflow between the beginning and 120 min low-flow ischemia. The electrical field stimulation-induced overflow of noradrenaline during hibernation myocardium was significantly less than preischemia or after reperfusion, but there was nosignificant difference between preischemia and reperfusion group. Tyramine induced significant noradrenaline release in absence of desipramine after 30 min reperfusion, but this increase in noradrenaline release had nosignificant in the presence of desipramine. These studies indicated that there was not significant spontaneous noadrenaline overflow during acute myocardial hibernation in isolated rat hearts, the stimulation-induced noradrenaline overflow decreased during hibernation and restored to the level of preischemia after reperfusion, myocardial noradrenaline response to tyramine remained after 30 min reperfusion.Conclusion: Myocardial noradrenaline overflow may not contribute to the development of acute myocardial hibernation and the function of sympathetic nerve may also maintain in hibernation as myocardium does during acute myocardial hibernation, reperfusion of myocardium may contribute to restoring the function of sympathetic nerve.

    • Lithium decreased NR2B tyrosine phosphorylation and interactions of NR2B and PSD-95 with Src in rat hippocampus following cerebral ischemia

      2005, 19(5). DOI: 10.7655

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      摘要:Objective: To study the effects of chronic lithium on N-methyl-d-aspartate receptor subunit 2B (NR2B) tyrosine phosphorylation and the interactions of NR2B and PSD-95 with Src induced by cerebral ischemia/reperfusion (I/R). Methods: Transient (15 min) cerebral ischemia was induced by four-vessel occlusion procedure in SD rats. Immunoprecipitation (IP) and immunoblotting (IB)were performed to investigate the phosphorylation and interactions of proteins. The effects of lithium on tyrosine phosphorylation of NR2B and its interactions with PSD-95 and Src were examined. Results: Transient cerebral ischemia 15 min followed by reperfusion 6 h (I/R 6h) caused a significant increase in tyrosine phosphorylation of NR2B. Administration of LiCl for 7days before ischemia caused a profound decrease in tyrosine phosphorylation of NR2B. Similiarly, the interactions of NR2B and PSD-95 with Src were also enhanced by I/R 6 h.moreover, these interactions were also inhibited by chronic lithium. Conclusion: Pretreatment with lithium decrease tyrosine phosphorylation of NR2B and interactions of NR2B and PSD-95 with Src during cerebral I/R.

    • The clinical application of multi-slice spiral CT angiography in abdominal aortic disease

      2005, 19(5). DOI: 10.7655

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      摘要:Objective: To evaluate the clinical application of multi-slice spiral CT angiography(MSCTA) in the assessment of abdominal aortic disease. Methods: Fifty-four patients underwent multi-slice spiral CT angiography of abdomen. Contrast agent (Omnipaque 300 I g/L) 1.5 ml/kg was injected and the injection rate was 3 ml/s. The delay time was determined by bolus tracking technique,Tll level abdominal aorta was set as the target vessel and the threshold was 180-200 Hu, slice width was 3 mm and with a pitch of 4-6.Original data were transferred to working-station to perform functional reconstruction. Results: Ten cases were normal, twenty-eight cases were abdominal aortic aneurysms, five abdominal aortic dissecting aneurysms (Debakay type Ⅲ ) and eleven aortic sclerosis. SSD showed the body of aneurysm and the relationship between aneurysm and adjacent blood vessel, MIP better displayed calcification of blood vessel wall and condition of the stent, MPR demonstrated true and false lumen, rupture site of abdominal aorta intima and mural thrombus. Conclusion: MSCTA axial and reconstruction image can show the extent of abdominal aortic disease and the relationship with adjacent blood vessels. It is a safe, simple and non-invasive examination method.

    • Radiological analysis of image on geriatric pulmonary tuberculosis

      2005, 19(5). DOI: 10.7655

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      摘要:Objective: To evaluate the radiological features in the cases with geriatric pulmonary tuberculosis(GPTB). Methods: One hundred and two elderly patients with tuberculosis were studied in this paper. All patients had an examination on chest X-ray, and 63 of them had a chest scan of CT. Results: The pulmonary tuberculosis infiltration, caseous pneumonia and cavitary lesions were found in 85, 11 and 19 cases with GPTB respectively. Acute miliary tuberculosis was diagnosed in 2 cases. Pleural effusion was in 26 cases. In addition, Hilar and mediastinal lymphadenopathy was presented in the other 13 cases. Conclusion: Radiological findings of GPTB were atypical in general.

    • Evaluation of speech function on repairing defects of maxilla and palate with temporalis muscle flap

      2005, 19(5). DOI: 10.7655

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      摘要:Objective: To evaluate the speech function on repairing defects of maxilla and palate with temporalis muscle flap after benign or malignant turmor resection. Methods: The lateral cephalogram and speech intelligibility were detected in 19 cases with the operations of repairing defects of maxilla and palate by temporalis muscle flap, and their recovery of the speech function were analyzed.Results: Among the 19 patients, there were 15 cases (78.00%) with complete velopharynx, 3 cases (15.80 % ) with mariginal velopharynx, and 1 case (5.26%) with insufficient velopharynx. The average speech intelligibility was 94.3%, close to the normal speech intelligibility. Conclusion: The operation of repairing defects of maxilla and palate with temporalis muscle flap can reconstruct the phonatory structure, preserve the palate function and restore the speech function after operation.

    • Anesthetic considerations for the patients with off-pump coronary artery bypass grafting

      2005, 19(5). DOI: 10.7655

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      摘要:New surgical techniques for coronary artery bypass graft (CABG) have been explored in order to minimize disfiguring scars, avoid cardiopulmonary bypass and decrease hospital residence and costs etc. Improved surgical expertise can perform the procedures in some high-risk patients such as poor left ventricular function, redo CABG, advanced age and renal dysfunction etc. Intraoperatively there is an obligatory period of myocardial ischemia, which may lead to significant hemodynamic consequences. It is a quite challenge to anesthetist in managing the intraoperative patient.