细胞角质素CK18和CK19在肝细胞癌组织中的表达

目的:观察和探讨细胞角质素CK18和CK19在不同肝组织中的表达及其意义.方法:采用免疫组化SP法观察正常肝组织8例.肝硬化组织27例和肝细胞癌(hepatocellular carcinoma,HCC)组织43例中CK18,CK19的表达.结果:肝CK18和CK19在肝硬化组与正常肝组织的表达率均无明显差异,但两者在肝细胞癌组与肝硬化组之间差异有显著性(65.1%vs29.6%.69.8%vs 25.9%,P＜0.01).在20/27例肝硬化组织和35/43例HCC组织中可见到CK18,CK19标记的卵圆细胞,且在两种组织中卵圆细胞的数量有显著性差异(CK18:6.57-4-1.69vs 10.70±2.31:CK19:5.37±1.17vs10.45±2.15,P＜0.01).结论:CK18和CK19参与了肝硬化到HCC的癌变过程;CK18和CK19在卵圆细胞中的强阳性表达,支持HCC的卵圆细胞起源学说.     

黄芪汤对肝硬化大鼠肝脏卵圆细胞肝向分化的作用

目的:研究黄芪汤对肝硬化大鼠肝脏卵圆细胞肝向分化的作用机制。方法:应用二甲基亚硝胺制备大鼠肝硬化模型,用黄芪汤治疗,进行肝功能、肝脏病理学检测;应用共聚焦免疫荧光技术检测肝脏卵圆细胞与肝细胞及胆管细胞共定位染色。结果:黄芪汤能显著降低模型大鼠血清AST、TBil水平和肝组织Hyp含量,减少肝组织α-SMA的表达,在模型大鼠肝纤维化逆转过程中,黄芪汤可使Thy1.1与CK19共定位细胞数量显著增加,使肝脏卵圆细胞(HOC)表型和功能发生改变。结论:黄芪汤通过诱导肝脏卵圆细胞肝向分化作用来促进肝硬化逆转。     

Twist、E-cadherin在胆管细胞癌中的表达及意义

目的:探讨转录因子Twist及皮黏附分子E-cadherin在人肝内外胆管细胞癌、癌旁和正常胆管中的表达及其意义.方法:收集2008-01/2009-03华中科技大学同济医学院附属协和医院肝胆外科和胰腺外科中心手术切除的胆管细胞癌患者49例,其中肝内胆管细胞癌(intrahepatic cholangiocarcinoma,ICC)29例.肝外胆管细胞癌(extrahepatic cholangiocarcinoma,ECC)20例:另取24例癌旁组织(ICC与ECC各12例)、20例正常的胆管组织作为对照.免疫组织化学法检测Twist及E-cadherin的表达,分析其临床病理意义.结果:Twist蛋白在ICC及ECC中表达明显强于癌旁组织或正常胆管(79.31%,80.0%vs20.83%,1 5.0%,均P<0.05):E-cadherin在ICC及ECC中表达明显弱于癌旁组织或正常胆管组织(44.83%,40.0%vs 95.24%,100%,均P<0.05),组间表达差异有显著性:Twist和E-cadherin在胆管细胞癌中表达呈高度负相关(r=0.67,P<0.01);Twist蛋白表达水平与年龄、性别及组织病理分级无关(P>0.05).而与CA19-9水平、肿瘤浸润及淋巴结转移有关(P<0.01).结论:Twist与E-cadherin的异常表达与胆管细胞癌的发生发展相关;Twist可能参与了胆管细胞癌浸润转移的过程.     

肝细胞癌中DcR3表达与卵圆细胞增殖的可能关系

目的研究DcR3（一种新的凋亡相关分子）在肝细胞癌（HCC）中表达情况及其与卵圆细胞增殖之间可能存在的关系,以探讨卵圆细胞增殖活化的分子机制。方法对52例HCC组织,用免疫组织化学EnVision法同时标记肝细胞分化标记物hepatocyte paraffinl（HEP）、胆管标记物CK7/CK19、细胞增殖标记物Ki-67以及DcR3,半定量分析卵圆细胞形态数量和凋亡细胞数量的关系。结果在所有HCC肿瘤旁硬化组织的反应性小胆管中,均可见CK7、CK19和DcR3的表达,而在52例HCC中有23例见部分肿瘤细胞表达CK7和/或CK19,36例部分肿瘤细胞表达DcR3。其中23例CK7和/或CK19阳性病例中,21例同时有DcR3阳性（91.3%）,而29例CK7/CK19阴性病例中仅有15例（51.7%）DcR3阳性表达,两组间差异有统计学意义（P=0.002）。结论HCC中存在CK7和/或CK19阳性表达的卵圆细胞,表明卵圆细胞在部分HCC的发病中起重要作用,而这些病例多数有DcR3高表达,同时可见DcR3高表达于癌旁硬化组织反应性小胆管中,这些均提示DcR3的抗凋亡作用可能与卵圆细胞的增殖分化有关。     

大鼠酒精性肝病细胞凋亡与细胞色素P4502E1和氧化应激的关系

目的:观察大鼠酒精性肝病组织病理形态学改变,探讨细胞凋亡与细胞色素P4502E1的表达以及和氧化应激的关系.方法:用酒精灌胃法制备酒精性肝病大鼠模型,模型组给予酒精8 g/kg,每天分2次灌胃连续8 wk,对照组给予等量的生理盐水灌胃.实验8 wk末,观察肝组织的病理形态学改变,用原位末端标记法(TUNEL)检测肝细胞凋亡,用免疫组化法检测肝组织中Caspase-3蛋白表达,用全自动生化仪检测ALT和AST的含量,用PCR法测定肝细胞色素P4502E1的基因表达,分别用硫代巴比妥酸法(TBA法)和黄嘌呤氧化酶法测定肝组织丙二醛(MDA)的含量和超氧化物歧化酶(SOD)的活力.结果:模型组凋亡的肝细胞明显增多,主要分布在中央静脉周围、点状和灶状坏死区;Caspase-3主要分布于中央静脉及肝细胞坏死灶周围细胞的胞质中.模型组肝细胞凋亡指数(AI)和Caspase-3蛋白表达强度明显高于对照组(AI:6.2%±1.7% vs 1.7%±0.8%;Caspase-3:66.7% vs 9.5%,P<0.05,P<0.01).CYP2E1表达:对照组c1基因频率为91.6%,c2基因频率为8.4%;模型组c1基因频率为53.4%,c2基因频率为46.6%,均有显著性差异(P<0.05).长期酒精摄入大鼠血清MDA含量增加(41.53±7.43μmol/L vs 15.72±2.06μmol/L,P<0.05),SOD活力下降(353.12±61.02 kU/L vs 636.82±138.60 kU/L,P<0.05),与酒精性肝病肝细胞凋亡程度有相关性(r=0.644,r=-0.511).结论:长期酒精摄入可引起大鼠酒精性肝病及及肝功能损伤,肝细胞凋亡明显增加.CYP2E1基因PstⅠ及RsaⅠRFLPs与酒精性肝病有关,其中c2基因可能与大鼠酒精性肝病的发生有关.MDA含量和SOD活力在酒精性肝病的肝细胞凋亡过程及脂质过氧化反应中发挥重要作用.     

原发性肝癌组织芯片中乙酰肝素酶的表达及意义

目的:探讨乙酰肝素酶(heparanase,HPA)蛋白在原发性肝细胞癌(HCC)组织芯片中的过度表达及临床意义.方法:125例HCC患者肝组织、48例肝癌患者癌旁组织、62例肝硬化患者肝组织及23例肝血管瘤患者相应正常肝组织构建组织微阵列.应用免疫组织化学检测HPA蛋白的表达水平,并分析其与HCC临床病理特征的关系.结果:HCC组织中的HPA蛋白的阳性率45.83%明显高于癌旁组织27.08%(x~2=2.23,P<0.05),肝硬化6.45%(x~2=5.262,P<0.05)和正常肝组织4.35%(x~2=3.895,P<0.05).癌旁组织中的HPA蛋白阳性率明显高于肝硬化(x~2=2.882,P<0.05)及正常肝组织(x~2=2.361,P<0.05);HCC中临床TNM分期ⅠⅡ期HPA阳性率明显低于ⅢⅣ期(29.41% vs 67.31%,x~2 =4.111,P<0.05);HCC中无转移组HPA阳性率明显低于转移组(14.71% vs 63.33%,x~2= 3.978,P<0.05);HPA表达率在AFP≥400μg/L和AFP<400μg/L组(52.05% vs 36.17%,x~2= 2.071,P<0.05)、有无门脉癌栓组(71.74% vs 29.73%,x~2=4.472,P<0.05)、多个和单个肿瘤结节组(73.91% vs 28.38%,x~2=4.847,P<0.05)以及肿瘤直径≥5 cm和<5 cm组(57.89% vs 25%,x~2=3.471,P<0.01)分别具有显著性意义.HPA表达与年龄、性别、分化程度、有无肝硬化及肿瘤包膜浸润无关.结论:HPA高表达在HCC的发生、发展及转移中起重要作用.检测HPA蛋白指标有助于HCC诊断和判断患者预后.     

过渡型CK19阳性表达细胞的研究

目的探讨慢性乙型肝炎患者肝组织过渡型CK19阳性表达细胞与肝癌前病变的关系。方法用LSAB免疫组织化学染色方法观察慢性乙型肝炎患者肝组织CK19的表达，每3个月对患者检查B超和血清甲胎蛋白，随访至1年。结果肝组织出现卵圆细胞分化为肝细胞的过渡型CK19阳性表达细胞的患者，大多数出现血清甲胎蛋白的明显升高。随访至1年，有1例患者出现小肝癌，1例出现肝实质低回声结节。结论过渡型CK19阳性表达细胞有可能作为一种新的肝癌前病变的病理学标志。     

CK-19与GPC-3联合检测在肝细胞癌和胆管细胞癌诊断中的价值

目的探讨免疫组化联合检测细胞角蛋白-19(cytokeratin-19,CK-19)及磷脂酰肌醇蛋白多糖-3(glypican-3,GPC-3)在肝细胞癌和胆管细胞癌诊断中的价值。方法采用CK-19及GPC-3免疫组化法对85例肝细胞癌组织和72例胆管细胞癌组织进行检测,比较CK-19及GPC-3在不同组织中的表达情况。结果在GPC-3免疫组化检测中,肝细胞癌组织中检测出75例阳性,阳性表达率高达88.24%,且中低分化组织阳性表达率明显高于高分化组织(P0.01)。而胆管细胞癌组织中仅有11例表达阳性,阳性表达率为15.28%,两种组织阳性表达率比较差异有统计学意义(P0.01)。在CK-19免疫组化检测中,胆管细胞癌组织呈现高阳性表达,表达率为94.44%,且中低分化组织阳性表达率明显高于高分化组织,而肝细胞癌表达均为阴性,与胆管细胞癌阳性表达率比较差异有统计学意义(P0.01)。结论GPC-3在肝细胞癌中呈现高表达,而CK-19仅在胆管细胞癌表达,联合检测CK-19与GPC-3对鉴别诊断肝细胞癌和胆管细胞癌有参考价值。     

蛋白激酶CK2β在肝细胞癌中的表达及临床意义

目的 研究蛋白激酶CK2β在肝细胞癌组织中的表达及临床意义,并探讨其表达与患者预后的关系。方法 收集2012年1月-2013年6月于郑州大学第一附属医院肝胆胰外科确诊的127例肝细胞癌患者癌组织及癌旁组织,采用免疫组化法检测其CK2β的表达,分析癌组织中CK2β表达强度与肝癌患者临床特征的关系。另收集本院2018年3-6月期间手术切除的20例肝细胞癌癌组织、癌旁组织、肝硬化组织、正常肝组织标本并提取蛋白和mRNA,运用Westen Blot和real-time PCR检测其CK2β表达情况。多组间均数比较采用单因素方差分析,进一步两两比较采用Bonferroni检验,计数资料组间比较采用χ2检验。CK2β在肝细胞癌的表达差异与临床特征参数的关系采用Mann-Whitney U检验或Kruskal-Wallis H检验。生存分析采用Kaplan-Meier法,组间比较采用log-rank检验。结果 Westen Blot和real-time PCR结果证实CK2β在癌组织中表达率显著高于癌旁组织、肝硬化组织和正常肝组织(P值均0. 05);癌旁及肝硬化组织中CK2β表达无差异(P值均 0. 05),且均高于正常组织表达(P值均0. 05)。免疫组化染色结果发现127例肝细胞癌患者的癌组织和癌旁组织中CK2β阳性表达率分别为85. 8%和63. 0%,差异有统计学意义(P 0. 001)。肝癌组织中CK2β的表达分布在不同年龄(Z=-2. 277,P=0. 023)、肝硬化有无(Z=-2. 144,P=0. 032)、不同肿瘤大小(Z=-2. 289,P=0. 004)、不同Edmondson-Steiner病理分级(χ2=8. 210,P=0. 016)的肝细胞癌患者中差异均有统计学意义。Kaplan-Meier生存曲线发现,CK2β表达强阳性组术后生存期均明显低于中阳性组、弱阳性组及阴性组(P值均0. 001)。结论CK2β蛋白可能参与肝细胞癌的发生发展,其阳性表达与肝细胞癌患者预后有关。     

胆汁性肝硬化大鼠肾细胞凋亡及其机制

目的:探讨胆汁性肝硬化大鼠肾脏病理组织学改变、肾脏细胞凋亡及其机制.方法:应用胆总管结扎(CBDL)技术制备大鼠胆汁性肝硬化模型.将SD大鼠随机分为:假手术组(SO)、CBDL 2 wk组与5 wk组.显微镜下观察肝脏和肾脏组织学改变;生化法测定内生肌酐清除率(Ccr),比色法测定肾组织丙二醛(MDA)含量,TUNEL法检测肾组织细胞凋亡,免疫组化测定Bcl-2,Bax蛋白的表达.结果:CBDL大鼠2 wk发生肝纤维化,5 wk则形成肝硬化;5 wk时肾小管上皮细胞有脱落、坏死;电镜下基底膜不规则增厚,系膜增宽,上皮细胞足突融合.模型大鼠5 wk组Ccr明显降低,较2 wk组与SO组有显著性差异(1.10±0.03vs 1.40±0.03,1.80±0.02,P＜0.05或0.01),肾组织丙二醛5 wk组较2 wk组与SO组显著性增高(4.07±0.51vs 2.32±0.85,0.79±0.37,均P＜0.01);模型大鼠5 wk组肾组织凋亡指数显著性高于2 wk组和SO组(68.36%±8.71%vs 24.08%±2.59%,11.43%±2.77%;均P＜0.01). SO组、2 wk组及5 wrk组Bcl-2逐渐下降,Bax表达则逐渐升高,各组间有显著差异(p＜0.05).结论:胆汁性肝硬化大鼠存在肾病理组织学改变与细胞凋亡:凋亡抑制蛋白Bcl-2低表达和凋亡诱导蛋白Bax高表达在肾损害中起作用.     

Medical and Psychology Students' Knowledge and Attitudes Regarding Aging and Sexuality

The current study surveys medical and doctoral psychology students (N = 100) from an urban northeastern university regarding knowledge and attitudes toward elderly sexuality and aging using the Facts on Aging Quiz, the Aging Sexuality Knowledge and Attitudes Scale, and measures of interest in gerontology, academic/clinical exposure to aging and sexuality, and contact with elders. The current study found that psychology students demonstrated greater aging knowledge than medical students; however, both groups showed gaps in knowledge about sexuality. Married students had greater academic/clinical exposure and greater knowledge about aging but less permissive attitudes toward elderly sexuality. Generally, knowledge about aging was the strongest correlate of knowledge about sexuality. Level of knowledge about sexuality was not associated with attitudes. Attitudes toward sexuality and aging may be more strongly tied to demographic variables reflective of religious beliefs or adherence to sociocultural norms.     

Leptin and cellular and innate immunity in abstinent alcoholics and controls

BACKGROUND: Basic studies indicate that in vitro and in vivo doses of leptin modulate cellular immune responses. Given evidence that concentrations of leptin are altered in alcoholics who also show immune abnormalities, this study examined the relationships between circulating levels of leptin and markers of cellular and innate immunity. METHODS: Circulating levels of leptin, natural killer cell (NK) activity, interleukin-2 (IL-2)-stimulated NK activity, and concanavalin A-stimulated production of IL-2, IL-6, IL-10, and IL-12 were compared between abstinent DSM-IV alcohol-dependent men (n = 27) and age- and gender-matched controls (n = 34). RESULTS: As compared with controls, alcoholics showed lower NK activity (p < 0.01) and a trend for lower levels of leptin (p = 0.055). In the total sample, leptin predicted NK activity (beta = 0.33; p < 0.05) after controlling for the confounding influence of body mass index, alcohol intake, and smoking. Leptin was not correlated with any of the cytokine measures. To examine whether the effects of leptin were mediated by its direct action on NK, additional studies examined in vitro effects of leptin on NK activity in healthy volunteers (n = 10); leptin doses (0.1, 1, and 10 nM) yielded levels of NK activity comparable to those with media alone. CONCLUSIONS: These data show that circulating levels of leptin are associated with NK activity in humans and suggest that abnormal in vivo concentrations of leptin may contribute to the declines of NK activity in alcoholics who are at risk for infectious diseases.     

自身免疫与动脉粥样硬化的发生发展

在动脉粥样硬化的发生发展过程中,自身免疫具有重要作用,本文就近年有关动脉粥样硬化的免疫机制的研究进展作一简要综述.     

Cement and Concrete Nanoscience and Nanotechnology

Concrete science is a multidisciplinary area of research where nanotechnology potentially offers the opportunity to enhance the understanding of concrete behavior, to engineer its properties and to lower production and ecological cost of construction materials. Recent work at the National Research Council Canada in the area of concrete materials research has shown the potential of improving concrete properties by modifying the structure of cement hydrates, addition of nanoparticles and nanotubes and controlling the delivery of admixtures. This article will focus on a review of these innovative achievements.     

慢性饮酒、吸烟与骨质疏松的相关性研究

目的 探讨长期大量慢性饮酒、吸烟等不良生活方式与骨质疏松的关系.方法 以乏力、腰背痛、双下肢酸困为主诉,且有长期大量慢性饮酒史、吸烟史,无其他慢性疾病史的年轻男性患者45例为观察对象,测定其血钙(Ca)、血磷(P)和血肌酐(Cr)、尿素氮(BUN)、血糖(Glu)、天门冬酸氨基转移酶(AST)、丙氨酸基转移酶(ALT)、血清碱性磷酸酶(ALP)、甲状腺功能、全段甲状旁腺激素(iPTH)、1,25-二羟维生素D3[1,25-(OH) 2D3]、双能X线骨密度仪测定腰椎L1-4及左侧股骨近段(包括颈、股骨颈上部、大粗隆)骨密度(BMD),分析饮酒、吸烟对骨量的影响.以感冒发热就诊、年龄相当、无饮酒、吸烟史及慢性疾病史的45例男性为对照组.结果 观察组1,25(OH)2D3水平均降低,血钙水平偏低,血磷正常,碱性磷酸酶呈不同程度地升高,骨密度T值提示观察组27例骨质疏松,12例骨量减少,6例正常骨量;而对照组仅有6例为骨量减少,1例骨质疏松,38例为正常骨量,两组比较差异有统计学意义(P＜0.05).结论 长期慢性大量饮酒吸烟影响骨量,甚至易导致骨质疏松.     

Immune and serologic profiles of HIV-infected and noninfected hemophilic children and adolescents

Objectives: To assess relationships among the effects of HIV on hemophilic children and adolescents' immunologic parameters and vaccine-related serology. Methods: We analyzed data from extensive baseline immunologic evaluations of 207 HIV antibody-positive (HIV+) and 126 HIV antibody-negative (HIV?) hemophilic children and adolescents. Results: HIV+ and HIV- participants differed significantly in T-lymphocyte subpopulation numbers, immunoglobulin levels, and seroprevalence rates for diphtheria toxoid, measles, and mumps antigens. IgG levels, IgM levels, and serologic titers to vaccine antigens showed little correlation with T-cell parameters. Proportionately more HIV+ participants were nonreactive to each and all of a panel of 7 skin test antigens (71 % vs 28% anerglc, RR 2.6). The odds of anergy increased 1.6 times for every decline of 200 CD4 ± cells/μTl. Conclusions: HIV had significant, largely independent T- and B-lymphocyte effects on this pediatric cohort. © 1994 Wiley-Liss, Inc.     

ADH and ALDH Polymorphisms and Alcohol Dependence in Mexican and Native Americans

Background: Ethanol is primarily metabolized in the liver by two rate-limiting reactions: conversion of ethanol to acetaldehyde by alcohol dehydrogenase (ADH) and subsequent conversion of acetaldehyde to acetate by aldehyde dehydrogenase (ALDH). ADH and ALDH exist in multiple isozymes that differ in their kinetic properties. Notably, polymorphisms within the genes that encode for these isozymes vary in their allele frequencies between ethnic groups, and thus, they have been considered as candidate genes that may differentially influence risk for the development of alcohol dependence across ethnic groups. Objectives and methods: Associations between alcohol dependence and polymorphisms in ADH1B, ADH1C, and ALDH2 were compared in a community sample of Native Americans (n 791) living on reservations and Mexican Americans (n 391) living within the same county. Results: Two Mexican Americans and no Native Americans possessed one ALDH2*2 allele. Presence of at least one ADH1B*2 allele was found in 7% of the Native Americans and 13% of the Mexican Americans, but was only associated with protection against alcohol dependence in the Mexican Americans. Presence of at least one ADH1B*3 allele was found in 4% of the Native Americans and 2% of the Mexican Americans, but was associated with protection against alcohol dependence only in the Native Americans. No associations between alcohol dependence and polymorphisms in ADH1C were found. Conclusions and Scientific Significance: Polymorphisms in ADH1B are protective against alcoholism in these two populations; however, these findings do not explain the high prevalence of alcoholism in these populations.