食管鳞状细胞癌组织中酪氨酸激酶受体B和脑源性神经生长因子蛋白及mRNA的表达及意义

目的 探讨酪氨酸激酶受体B( TrkB)和脑源性神经生长因子(BDNF)蛋白及mRNA在食管鳞状细胞癌(ESCC)组织中的表达及其意义.方法 应用免疫组织化学及原位杂交法检测59例ESCC、27例癌旁不典型增生组织及36例正常食管黏膜组织中TrkB和BDNF蛋白及mRNA的表达.结果 ESCC组织中TrkB蛋白及mRNA的阳性表达率分别为71.2％和64.4％,显著高于癌旁不典型增生组织(阳性率分别为48.1％和33.3％)及正常食管黏膜组织(阳性率均为0.0％),组间比较差异有统计学意义(P＜0.05);此外,BDNF蛋白和mRNA在ESCC组织中的阳性表达率分别为76.3％和69.5％,也显著高于癌旁不典型增生组织(阳性率分别为55.6％和40.7％)及正常食管黏膜组织(阳性率均为0.0％),组间比较差异有统计学意义(P＜0.05).TrkB和BDNF蛋白及mRNA的表达均与ESCC的分化程度、浸润深度和淋巴结转移密切相关(P＜0.05).进一步相关分析结果显示,TrkB mRNA和蛋白的表达均与BDNF mRNA和蛋白的表达呈正相关(P＜0.05).结论 TrkB和BDNF蛋白及mRNA表达与ESCC的发生、发展和转移密切相关.     

Ezrin在胃癌组织中的表达及其临床意义

目的 探讨Ezrin在胃癌发生及发展过程中的作用.方法 收集2008年6月至2009年5月第三军医大学西南医院全军普通外科中心收治的60例胃癌患者的胃癌组织及正常胃黏膜组织标本,分别采用RT-PCR和Western blot法检测标本中Ezrin mRNA及蛋白的表达情况,分析Ezrin与患者性别、年龄、肿瘤分化程度、病理分期、浸润深度及淋巴结转移间的关系.采用t检验、x2检验、Spearman等级相关检验进行统计学分析.结果 60例正常胃黏膜组织中有33例(55％) Ezrin mRNA表达水平升高,45例(75％)Ezrin蛋白表达水平升高;60例胃癌组织中有21例(35％) Ezrin mRNA表达水平升高,22例(37％) Ezrin蛋白表达水平升高.Ezrin mRNA及蛋白在正常胃黏膜组织中的表达水平分别为1.30±0.04和3.57±0.45,在胃癌组织中的表达水平分别为0.53±0.36和0.96±0.18,两者比较,差异有统计学意义(t=5.309,22.617,P＜0.05).胃癌组织中Ezrin mRNA及蛋白表达水平呈正相关(r=0.602,P＜0.05).Ezrin mRNA和蛋白表达在胃癌不同病理分期、不同浸润深度、有无淋巴结转移方面差异有统计学意义(x2=6.41,6.49,4.62;5.40,8.87,4.12,P＜0.05),而在性别、年龄和分化程度方面差异无统计学意义(x2=0.50,0.07,1.07;0.01,1.16,1.96,P＞0.05).结论 胃癌组织中Ezrin表达水平降低,这可能是胃癌的发生、发展及转移的机制之一.     

蛋白激酶激活受体-2在人类结直肠癌中表达的研究

目的研究蛋白酶激活受体-2(PAR-2)在结直肠癌中的蛋白表达和mRNA表达的水平,并探讨其临床价值。方法采用免疫组织化学法、免疫荧光分析法和实时荧光定量PCR的方法,检测38例结直肠肿瘤患者的正常肠黏膜组织、癌旁组织和癌组织PAR-2的蛋白和mRNA的表达情况。结果(1)PAR-2在结直肠正常肠黏膜、癌旁组织和癌组织中表达阳性率分别为25%、45%和85%;(2)PAR-2的表达强度的IOD值由弱到强依次为正常肠组织(681±101)、癌旁组织(6627±971)和癌组织(15861±2709),三组间经方差分析差异有统计学意义(F=132.6,P<0.05);(3)PAR-2 mRNA在正常肠组织、癌旁组织和癌组织的表达水平分别为1.57±0.69、2.91±1.4和4.09±1.99,三组间的差异有统计学意义(F=9.688,P=0.00);结论PAR-2的表达在结直肠肿瘤中高表达;PAR-2的表达和结直肠肿瘤的发生和发展密切相关。     

Survivin mRNA和蛋白在肝母细胞瘤组织中的表达

目的 观察肝母细胞瘤组织中Survivin mRNA与蛋白表达.方法 采用逆转录-聚合酶链反应(RT-PCR)方法检测16例肝母细胞瘤及相应癌旁组织中Survivin mRNA的表达,免疫组织化学SP法及Western blot法检测Survivin蛋白在肝母细胞瘤中的表达.结果 在肝母细胞瘤及其相应的癌旁组织中Survivin mRNA表达的阳性率分别为62.5％和11.1％,Survivin蛋白表达的阳性率分别为81.3％和22.2％,肝母细胞瘤组均远高于癌旁组,差异有统计学意义(P＜0.01);Survivin蛋白在肝母细胞瘤组织中的表达随肿瘤分期增高而增加;Ⅲ、Ⅳ期肝母细胞瘤组织Survivin mRNA 的表达量分别为2.390±0.071和4.506±0.309,明显高于早期肝母细胞瘤组织(P＜0.05).结论 Survivin 基因的高表达可能在肝母细胞瘤的发生及发展中起重要作用.     

PKM2和YAP在人肝癌组织中的表达及其相关性分析

目的探讨丙酮酸激酶M2(PKM2)和yes相关蛋白(YAP)在肝癌组织中的表达及其意义,并探索肝癌组织中PKM2蛋白和YAP蛋白表达的相关性。方法回顾性收集2010年4月至2013年10月期间于中国医科大学附属第四医院行手术切除的120例肝癌及其癌旁组织标本,采用SP免疫组织化学染色方法检测120例肝癌组织及癌旁组织PKM2和YAP蛋白的表达,并分析肝癌组织中PKM2蛋白和YAP蛋白表达与临床病理学特征的关系。其中,50例肝癌及癌旁组织标本具备手术时立即取材的新鲜冰冻组织标本,置液氮冷却后送-70°C冷冻冰箱内保存。采用Western blot与实时荧光定量PCR方法分别检测50例肝癌组织及癌旁组织中PKM2、YAP蛋白及其RNA的相对表达水平。结果 1免疫组化染色结果显示:肝癌组织中PKM2蛋白的表达阳性率为67.50%(81/120),YAP蛋白的表达阳性率为71.67%(86/120),均高于癌旁组织〔PKM2蛋白:20.83%(25/120);YAP蛋白:29.17%(35/120)〕,差异均有统计学意义(P0.050)。2肝癌组织中,PKM2蛋白和YAP蛋白的表达呈正相关(r=0.519,P0.001)。3肝癌组织中PKM2蛋白的表达与肿瘤直径、TNM分期、肿瘤分化程度及甲胎蛋白(AFP)水平均相关(P0.050),YAP蛋白的表达与肿瘤分化程度和AFP水平均有关(P0.050)。4 Western blot结果显示,肝癌组织中PKM2蛋白和YAP蛋白的表达水平分别为1.25±0.11和1.08±0.10,癌旁组织分别为0.38±0.01和0.41±0.02,肝癌组织中PKM2蛋白和YAP蛋白的表达水平均高于癌旁组织,差异均有统计学意义(P0.050)。5实时荧光定量PCR结果表明,以癌旁组织作为基准(即为1),肝癌组织中PKM2 m RNA和YAP m RNA的表达水平分别为11.38±0.35和19.96±0.48,均高于癌旁组织,差异均有统计学意义(P0.050)。结论 PKM2蛋白和YAP蛋白与肝癌的发生相关,并均与肝癌的分化程度密切相关。     

肾透明细胞癌组织中泛素连接酶Pirh2蛋白的表达及其临床意义

目的 探讨肾透明细胞癌组织中泛素连接酶Pirh2蛋白(p53-induced RING-H2 protein,Pirh2)的表达及与临床病理特征和预后的关系. 方法 肾透明细胞癌标本92例,G1 29例、G2 36例、G3 24例、G4 3例,T1 55例、T2 16例、T3 20例、T4 1例;正常肾组织标本20例作为对照组.采用免疫组织化学方法检测两组标本组织中Pirh2蛋白的表达,并结合临床资料进行分析. 结果 正常肾组织中Pirh2蛋白表达率为20.0％ (4/20),肿瘤组织为52.2％(48/92),且随组织病理分级和分期增加,Pirh2蛋白表达量逐渐增高.Pirh2蛋白在G1、G2、G3、G4组织中的高表达阳性率分别为24.1％、30.6％、75.0％和66.7％,T1、T2、T3、T4期肿瘤组织中高表达阳性率分别为25.5％、50.0％、75.0％和100.0％,分级间分期间差异有统计学意义(P=0.001).随访7～70个月,平均42个月,复发10例,复发组与未复发组中 Pirh2蛋白高表达阳性率分别为80.0％和36.6％,差异有统计学意义(P=0.022).Kaplan-Meier曲线显示,Pirh2蛋白高表达组与低表达组患者术后总生存率(71.1％与94.4％)和无复发生存率(78.9％与96.3％)比较,差异均有统计学意义(P＜0.01). 结论 Pirh2蛋白高表达与肾透明细胞癌的恶性程度及预后相关,Pirh2与肾透明细胞癌的发生和发展存在相关性.     

短发夹RNA干扰质粒与粉防己碱对结肠癌耐药细胞多药耐药性的逆转作用

目的 比较短发夹RNA干扰质粒与粉防已碱对结肠癌LoVo/5-氟尿嘧啶(5-FU)耐药细胞多药耐药性(MDR)的逆转作用.方法 用粉防己碱和靶向MDR1的短发夹RNA干扰质粒分别对结肠癌LoVo/5-FU耐药细胞进行处理,实验分为对照组(空白对照)、粉防己碱组和转染组(短发夹RNA干扰质粒).应用MTT法检测细胞对5-FU的敏感性;流式细胞仪检测细胞周期变化、凋亡情况及P-糖蛋白的阳性表达率;RT-PCR法检测MDR1 mRNA的表达情况;Western blot检测P-糖蛋白的表达情况.多组间的比较采用单因素方差分析,两两比较采用SNK-q检验.结果 (1)细胞对5-FU的敏感性:对照组、粉防己碱组、转染组的药物半数抑制浓度( IC50)分别为(7.3±0.3)、(4.4±0.7)、(2.4±0.4) mmol/L,3组比较,差异有统计学意义(F =65.27,P＜0.05);粉防己碱组与转染组比较,差异有统计学意义(q=6.67,P＜0.05).(2)细胞周期的变化:对照组、粉防己碱组、转染组G1期和S期细胞所占比例分别为38.13％±3.75％、51.36％±2.76％、59.24％±4.31％和20.46％±2.23％、14.32％±1.91％、9.40％±1.65％,3组比较,差异有统计学意义(F =25.23,24.37,P＜0.05);粉防己碱组与转染组比较,差异有统计学意义(q=3.67,4.35,P＜0.05).(3)细胞凋亡情况:对照组、粉防己碱组、转染组细胞凋亡率分别为1.32％±0.47％、3.24％±0.26％、5.88％±0.44％,3组比较,差异有统计学意义(F=99.26,P＜0.05);粉防己碱组与转染组比较,差异有统计学意义(q=11.48,P＜O.05).(4)P-糖蛋白的表达:对照组、粉防己碱组、转染组细胞P-糖蛋白的阳性表达率分别为96.9％±2.3％、61.6％±4.9％、76.6％±3.6％,3组比较,差异有统计学意义(F =67.83,P＜0.05);粉防己碱组与转染组比较,差异有统计学意义(q=6.97,P＜0.05).(5)MDR1 mRNA表达情况:对照组、粉防己碱组、转染组细胞MDR1 mRNA的相对表达量分别为1462±161、570±85、233±81,3组比较,差异有统计学意义(F=90.59,P＜0.05);粉防己碱组与转染组比较,差异有统计学意义(q=5.12,P＜0.05).结论 粉防己碱和靶向MDR1的短发夹RNA干扰质粒均能逆转LoVo/5-FU耐药细胞的MDR,且后者逆转作用更强,逆转机制与抑制P-糖蛋白表达及下调MDR1 mRNA表达有关.     

KAP-1对胰腺癌细胞Capan-2上皮细胞间质转化的促进作用

目的 研究KAP-1对胰腺癌细胞Capan-2上皮细胞间质转化(EMT)的调节作用.方法 构建LV-plenti-GFP-KAP-1慢病毒载体.将Capan-2细胞分为实验组(转染LV-plenti-GFP-KAP-1慢病毒载体)、阴性对照组(转染空载体)以及空白对照组1(含10％小牛血清的1640培养基);而后再将实验组Capan-2细胞分为抑制剂组(转染化学合成的miR-100-5p抑制剂)、空载体对照组(转染无关序列micro-RNAs抑制剂)、空白对照组2(含10％小牛血清的1640培养基).用双酶切及测序鉴定慢病毒并计算病毒滴度.LV-plenti-GFP-KAP-1慢病毒载体稳定转染至Capan-2细胞后,观察细胞的形态学改变.应用实时定量PCR检测各组细胞中KAP-1、EMT标志蛋白以及miR-100-5pmRNA的表达情况.应用Western blot法检测各组细胞中KAP-1、EMT标志蛋白表达水平.计量资料采用(x)±s表示,组间比较均采用方差分析.结果 成功构建LV-plenti-GFP-KAP-1慢病毒载体,病毒滴度为2×108 TU/ml.慢病毒载体转染Capan-2细胞48 h后,实验组细胞与对照组比较其形态有明显间质样改变.实验组、阴性对照组、空白对照组1细胞中各目标蛋白mRNA的相对表达量:KAP-1分别为1.77±0.83、5.03 ±0.29、5.13±1.14,N-钙黏蛋白分别为2.62 ±0.71、5.07 ±1.53、5.81 ±1.49,波形蛋白分别为2.50 ±0.21、3.83±0.57、4.92±0.90,E-钙黏蛋白分别为7.20±1.17、7.83 ±0.78、3.07±0.36,miR-100-5p分别为1.81 ±0.40、7.01 ±0.96、6.87±0.35,3组比较,差异有统计学意义(F=5.99,7.62,7.88,6.62,4.64,P＜0.05).抑制剂组、空载体对照组、空白对照组2细胞中KAP-1 mRNA的相对表达量分别为1.56 ±0.42、4.89 ±0.61、5.20 ±0.38,3组比较,差异有统计学意义(F=5.14,P＜0.05);波形蛋白mRNA的相对表达量分别为3.10±1.37、3.44 ±0.94、3.08±1.16,3组比较,差异无统计学意义(F=0.49,P＞0.05).Western blot检测结果表明:实验组、阴性对照组、空白对照组1细胞中各目标蛋白的相对表达量:KAP-1蛋白分别为2.77 ±1.99、0.83 ±0.46、0.71 ±0.26,N-钙黏蛋白分别为1.31 ±0.38、0.41 ±0.37、0.08 ±0.04,波形蛋白分别为4.25±0.63、1.03 ±0.33、1.37±0.92,E-钙黏蛋白分别为0.62 ±0.06、1.17 ±0.45、3.04±0.65,3组比较,差异有统计学意义(F=5.54,4.68,3.19,8.18,P＜0.05).抑制剂组、空载体对照组、空白对照组2细胞中KAP-1蛋白的相对表达量分别为2.27 ±0.71、0.56 ±0.43、0.61 ±0.39,3组比较,差异有统计学意义(F=4.81,P＜0.05);波形蛋白分别为3.19±0.55、3.93±0.06、3.61 ±0.73,3组比较,差异无统计学意义(F =0.04,P＞0.05).结论 KAP-1转录因子通过特异性调控其下游miR-100-5p表达,从而促进人类胰腺癌细胞Capan-2 EMT.     

MTA1表达与胸中段食管鳞癌预后的相关性

目的 探讨MTA1表达与胸中段食管鳞癌预后的相关性.方法 回顾2002年1月至2004年1月间,手术治疗165例胸中段食管癌病人临床资料.采用免疫组化进行肿瘤转移相关基因1(metastasis-associated gene 1,MTA1)检测,采用Kaplan-meier法进行生存分析、用Cox回归分析判定独立预后因素.结果 T2和T3病例MTA1表达率分别为13.3%和50%,两组差异有统计学意义(x2=13.2,P=0.00).有、无淋巴结转移者MTA1表达率分别为53.4%和31.2%,两组差异有统计学意义(x2=8.2,P=0.04).有、无MTA1蛋白表达者的5年生存率分别为18.3%和45.7%,两组差异有统计学意义(P=0.00).T2病例中,有、无MTA1蛋白表达者5年生存率分别为25.0%和50.0%,两组差异无统计学意义(P=0.20);T3病例中,有、无MTA1蛋白表达者的5年生存率分别为15.6%和40.6%(P=0.01),两组差异有统计学意义.pN0者中,有、无MTA1蛋白表达病例5年生存率分别为29.2%和54.7%(P=0.03),两组差异有统计学意义;pN1者中,有、无MTA1蛋白表达者5年生存率分别为12.8%和34.1%(P=0.04),两组差异有统计学意义.Cox回归分析结果显示,N分类和MTA1蛋白表达是独立的预后危险因素.结论 食管鳞癌不同的T、N分类中MTA1蛋白表达存在差别;MTA1蛋白过度表达者5年生存率降低;淋巴结转移和MTA1蛋白过度表达是独立的不利预后因素.     

Hedgehog信号通路在缺氧肠上皮屏障功能调控的作用

目的 探讨Hedgehog(Hh)信号通路在缺氧肠上皮屏障功能调控的作用.方法 大鼠小肠上皮细胞系IEC-6细胞分为3组:常氧组(21％氧浓度)、缺氧组(2％氧浓度)、缺氧+环巴胺组(用5 mmol/L的环巴胺预处理30 min后,再给予2％氧浓度进行缺氧处理).RT-PCR检测Hh信号通路IHH、PTCH、GLI-1 mRNA的表达变化,电阻测定仪检测跨上皮电阻(TER),Western blot检测IHH、紧密连接蛋白经典表达分子胞质附着蛋白(ZO-1)、咬合蛋白(Occludin)、闭合蛋白(Claudin-1)的表达情况.组间比较采取单因素方差分析,两两比较采用LSD-t检验.结果 RT-PCR检测结果表明:常氧组Hh信号通路IHH、PTCH、GLI-1 mRNA的相对表达量分别为0.056±0.009、0.459±0.087、0.142±0.023;缺氧组分别为0.303±0.052、0.678±0.073、0.483±0.061,两组比较,差异有统计学意义(t=-14.05,-11.85,-6.52,P＜0.05).Western blot检测结果显示:常氧组和缺氧组的IHH相对蛋白表达量分别为0.39±0.06和0.91±0.15,两组比较,差异有统计学意义(t=-8.08,P＜0.05).常氧组、缺氧组和缺氧+环巴胺组的TER分别为(134±5) Ohm/cm2、(100±6)Ohm/cm2、(118±5)Ohm/cm2,3组比较,差异有统计学意义(F=1.04,P＜0.05).与常氧组比较,缺氧组下降约27.7％(t=7.84,P＜0.05);与缺氧组比较,缺氧+环巴胺组回升约16.4％,但仍低于常氧组(t=4.23,P＜0.05).常氧组胞质附着蛋白-1、咬合蛋白、闭合蛋白-1的表达分别为1.18±0.24、0.80 ±0.13、0.90±0.09,缺氧组分别为0.58±0.08、0.32±0.05、0.50±0.09,缺氧+环巴胺组分别为0.92 ±0.21、0.43 ±0.10、0.82±0.11,3组比较,差异有统计学意义(F=4.95,2.88,10.09,P＜0.05).缺氧组较常氧组分别降低48.7％、40.0％、55.6％(t=12.86,9.35,18.90,P＜0.05);缺氧+环巴胺组较缺氧组分别提高59.9％、35.2％、65.1％(=5.63,2.92,6.66,P＜0.05).结论 缺氧条件可刺激激活Hh信号通路,从而引起紧密连接蛋白表达降低,导致肠上皮屏障功能损害.     

Presenteeism and Absenteeism Before and After Total Hip and Knee Arthroplasty

BackgroundAbsenteeism is costly, yet evidence suggests that presenteeism—illness-related reduced productivity at work—is costlier. We quantified employed patients’ presenteeism and absenteeism before and after total joint arthroplasty (TJA).MethodsWe measured presenteeism (0-100 scale, 100 full performance) and absenteeism using the World Health Organization’s Health and Work Performance Questionnaire before and after TJA among a convenience sample of employed patients. We captured detailed information about employment and job characteristics and evaluated how and among whom presenteeism and absenteeism improved.ResultsIn total, 636 primary, unilateral TJA patients responded to an enrollment email, confirmed employment, and completed a preoperative survey (mean age: 62.1 years, 55.3% women). Full at-work performance was reported by 19.7%. Among 520 (81.8%) who responded to a 1-year follow-up, 473 (91.0%) were still employed, and 461 (88.7%) had resumed working. Among patients reporting at baseline and 1 year, average at-work performance improved from 80.7 to 89.4. A Wilcoxon signed-rank test indicated that postoperative performance was significantly higher than preoperative performance (P < .0001). The percentage of patients who reported full at-work performance increased from 20.9% to 36.8% (delta = 15.9%, 95% confidence interval = [10.0%, 21.9%], P < .0001). Presenteeism gains were concentrated among patients who reported declining work performance leading up to surgery. Average changes in absences were relatively small. Combined, the average monthly value lost by employers to presenteeism declined from 15.3% to 8.3% and to absenteeism from 16.9% to 15.5% (ie, mitigated loss of 8.4% of monthly value).ConclusionAmong employed patients before TJA, presenteeism and absenteeism were similarly costly. After, employed patients reported increased performance, concentrated among those with declining performance leading up to surgery.     

The German emergency and disaster medicine and management systemdhistory and present

As well for optimized emergency management in individual cases as for optimized mass medicine in disaster management, the principle of the medical doctors approaching the patient directly and timely, even close to the site of the incident, is a long-standing marker for quality of care and patient survival in Germany. Professional rescue and emergency forces, including medical services, are the “Golden Standard” of emergency management systems. Regulative laws, proper organization of resources, equipment, training and adequate delivery of medical measures are key factors in systematic approaches to manage emergencies and disasters alike and thus save lives. During disasters command, communication, coordination and cooperation are essential to cope with extreme situations, even more so in a globalized world. In this article, we describe the major historical milestones, the current state of the German system in emergency and disaster management and its integration into the broader European approach.     

Work and career experiences and emotional well-being of managerial and professional women

This study examined a research model developed to understand emotional well-being among managerial and professional women. Data were collected from 792 women using questionnaires completed anonymously. Although considerable diversity was present in the sample, most women were in early career, married but still without children. Four groups of predictor variables identified in previous research were considered: personal demographic variables. Organizational and situational characteristics, work experiences associated with job and career satisfaction and work outcomes. Work experiences and work outcomes were fairly consistently and significantly related to self-reported emotional well-being. Implications for managerial women and their employing organizations are offered.     

Differences in Osteocyte Density and Bone Histomorphometry Between Men and Women and Between Healthy and Osteoporotic Subjects

Bone defects related to osteoporosis develop with increasing age and differ between males and females. It is currently thought that the bone remodeling process is supervised by osteocytes in a strain-dependent manner. We have shown an altered response of osteocytes from osteoporotic patients to mechanical loading, and osteocyte density is reduced in osteoporotic patients, which might relate to imperfect bone remodeling, leading to lack of bone mass and strength. Hence, information on osteocyte density will contribute to a better understanding of bone biology in males and females and to the assessment of osteoporosis. Osteocyte density as well as conventional histomorphometric parameters of trabecular bone were determined in cancellous iliac crest bone of healthy postmenopausal women and men and of osteoporotic women and men. Osteocyte density was higher in healthy females than in healthy males and lower in osteoporotic females than in healthy females. Bone mass was reduced in osteoporotic patients, both male and female. In females, trabecular number was reduced, whereas in males, trabecular thickness was reduced and eroded surface was increased. There were no correlations between the parameter groups bone architecture, bone formation, bone resorption, and osteocyte density. These results are consistent with impaired osteoblast function in osteoporotic patients and with a different mechanism of bone loss between men and women, in which osteocyte density might play a role. The reduced osteocyte numbers in female osteoporotic patients might relate to imperfect bone remodeling leading to lack of bone mass and strength. M. G. Mullender and S. D. Tan contributed equally to this work.