增殖细胞核抗原在大小鼠肠、肝和肾的表达

目的探讨增殖细胞核抗原(PCNA)在大小鼠肠、肝、肾的表达及其在大小鼠同一器官的表达差异。方法分别取体重为(111.6±16.0)gSD大鼠6只和(17.1±2.0)g昆明小鼠7只的肠、肝、肾,常规石蜡切片,苏木精伊红(HE)染色观察形态,SABC法检测PCNA的表达。结果PCNA在大小鼠肠绒毛固有层、肠腺细胞、肝细胞、肾小球和肾小管中有不同程度的表达,PCNA免疫反应阳性物质在肠、肝、肾的积分光密度在大小鼠同一器官的差异没有统计学意义(P>0.05)。结论正常情况下大小鼠肠、肝、肾均有一些细胞处于增殖状态,同一器官中PCNA的表达差异不明显。     

人ATP7B基因对Wilson病动物模型LEC大鼠暴发性肝炎的治疗

目的 探讨人ATP7B基因对Wilson病动物模型LEC大鼠暴发性肝炎的治疗效果。方法 将构建的7．1kb含有鸡β肌动蛋白启动子的人正常ATP7B cDNA，经显微注射法导人人Wilson病动物模型LEC(long-Evans Cinnamon)大鼠受精卵，建立转基因功能恢复大鼠模型。以Western bolt检测人ATP7B在转基因大鼠肝内的表达，用同窝无转基因大鼠及正常野生型LEA大鼠作对照，对6～16周龄的转基因大鼠的血清AST、ALT和总胆红素水平进行连续测定，同时取13周龄转基因大鼠的肝组织进行病理学和组织化学分析。结果 人ATP7B基因在转基因大鼠的肝组织中获得正确和完整的表达：12周龄前后，与同窝无转基因大鼠相比，转基因大鼠的血清AST、ALT和总胆红素水平明显降低，肝组织的炎性病变显著减轻，肝细胞内铜的蓄积减少；至16周龄，转基因大鼠的临床表型未见异常，存活率达100％。说明人ATP7B的导入，成功地抑制了Wilson病动物模型LEC大鼠的暴发性肝炎的发生。结论 Wilson病肝炎的发生与ATP7B基因功能缺陷引起的铜蓄积有直接关系。基因转移有可能是一种有效的治疗Wilson病的方法。     

大鼠肝癌发生过程中肿瘤坏死因子相关的凋亡诱导配体及核转录因子-κB的表达

目的:探讨肿瘤坏死因子相关的凋亡诱导配体(TRAIL)及核转录因子κB(NF-κB)在大鼠肝癌发生过程巾的表达及意义.方法:应用免疫组织化学SP法,对二乙基亚硝胺(DEN)诱发的大鼠肝癌发生过程中TRAIL及NF-κB的动态表达进行检测.结果:二乙基亚硝胺诱发的肝癌为肝细胞癌,大鼠肝癌癌变过程大致经过肝细胞损伤期、肝细胞增生-硬化期和肝细胞癌变期3个阶段.在正常大鼠肝、损伤期及增生硬化期偶见少量肝细胞呈TRAIL阳性表达,进入癌变期,TRAIL阳性细胞数量增多,主要位于癌结节内.在正常大鼠肝,偶见少量肝细胞呈NF-κB阳性表达,随着肝癌发生发展,阳性表达细胞逐渐增多,至诱癌晚期,可见大量NF-κB阳性表达细胞,均比正常肝组织表达高.结论:TRAIl及NF-κB可能与肝癌的发生发展密切相关.     

Hyper-IL-6重组腺病毒治疗大鼠急性肝衰竭的实验研究

目的 探讨重组腺病毒介导的超级白细胞介素6(Hyper-IL-6)对大鼠急性肝衰竭的治疗作用及机制.方法 80只急性肝衰竭模型大鼠随机分为:未感染组、AdO组、AdlL-6组、AdHIL-6组,分别予腹腔注射生理盐水、AdO、AdIL-6、AdHIL-6,后进行肝功能指标、肝脏病理学及肝细胞原位凋亡检测,并观察增殖细胞核抗原(PCNA)、caspase-3在肝组织中的表达情况及大鼠14 d存活率.结果 AdHIL-6组血清丙氨酸氨基转移酶(ALT)、总胆红素(TB)含量及凋亡指数明显降低,而肝组织PCNA表达明显增高(P<0.01),肝组织病变减轻,caspase-3表达阳性,14 d存活率较高.结论 Hyper-IL-6对急性肝衰竭大鼠有明显的促肝细胞增殖及抗肝细胞凋亡效应,能有效改善肝功能及肝组织学状况,提高肝衰竭大鼠存活率.     

层粘蛋白及其受体、增殖细胞核抗原与肝细胞癌肿瘤生物学特性的关系

目的:探讨层粘蛋白(LN)、层粘蛋白受体(LN-R)及增殖细胞核抗原(PCNA)与原发性肝细胞癌(HCC)恶性程度的关系.方法:采用SABC法分别检测42例肝细胞癌及癌旁组织中LN、LN-R及PCNA的表达情况.结果:LN、LN-R及PCNA在肝细胞癌中的表达均显著高于癌旁组织(P＜0.05),且与肝细胞癌Edmondson分级呈正相关;浸润型(IHCC)生长、有转移、肿瘤直径＞3cm者LN、LN-R及PCNA的表达高,而膨胀型(EHCC)生长、无转移、肿瘤直径≤3cm者多表达较低;在高增殖指数组LN、LN-R表达高于低增殖指数组(P＜0.05).结论:细胞内LN、LN-R及PCNA可作为评判肝细胞癌恶性程度及临床预后的良好指标.     

肝细胞癌组织中Glypican-3基因表达及其调控机制

目的：探讨肝细胞癌中Glypican-3（GPC3）基因表达及调控机制。方法:采用RT-PCR和聚合酶链反应-单链构象多态性(PCR-SSCP)方法分别检测48例肝细胞癌的癌组织、39例癌旁组织和31例正常肝组织中GPC3 mRNA的表达和基因突变；免疫组化S-P法检测GPC3、P53和PCNA蛋白的表达。结果:GPC3 mRNA在肝细胞癌的癌组织中的阳性表达率为77.1%，但其在癌旁和正常肝组织中不表达；肝细胞癌的癌组织中未发现GPC3基因突变；GPC3与P53蛋白表达无相关性（r＝-0.12574，P＞0.05），肝细胞癌的癌组织中GPC3 表达阳性和阴性的增殖细胞核抗原(PCNA)平均指数分别为（46.32±27.54）%、（39.83±21.47）%,P＞0.05。结论:肝细胞癌组织中GPC3 mRNA高表达与基因突变无关，没有直接影响P53和PCNA蛋白表达。     

可溶型B7-H3在原发性肝细胞癌患者血清中的表达及临床意义

目的 检测协同刺激分子B7-H3在肝细胞癌患者血清及组织中的表达,并探讨其临床意义.方法 选取原发性肝细胞癌63例患者的癌组织和血清标本;5例肝血管瘤的瘤旁组织作为正常肝组织对照;同期收取50例健康体检人员外周血标本为正常对照.采用ELISA检测肝癌患者和正常人血清中可溶型B7-H3(sB7-H3)的含量,免疫组化检测正常肝组织和肝细胞癌组织中B7-H3的表达.结果 肝细胞癌患者外周血清sB7-H3含量为(4143.47±976,27) pg/mL,显著高于正常对照组(2076.18±605.42)pg/mL(P ＜0.05);且其表达水平与临床分期,是否远处转移及肝癌组织中是否阳性表达B7-H3等参数有关(P＜0.05),与年龄、性别、组织学类型、淋巴结转移及肿瘤大小等临床病理参数无关;与血清学指标CA19-9呈明显正相关(P＜0.05),但与AFP和CEA水平无相关性.结论 肝癌患者血清中sB7-H3表达明显高于正常人,B7-H3表达与疾病临床病理指标相关,表明检测sB7-H3可能对原发性肝癌的诊断有一定价值.     

富硒螺旋藻对肝叶切除大鼠肝细胞再生的促进作用

目的: 探讨富硒螺旋藻对肝叶切除大鼠肝细胞再生的促进作用。方法: 用Wistar大鼠复制67%肝叶切除动物模型，术前灌胃富硒螺旋藻7 d，于手术时(0 h)及术后12 h、48 h取鼠肝细胞，检测谷胱苷肽过氧化物酶(GPx)和硫氧还蛋白还原酶(TR)活性，免疫组化方法测定肝细胞增生细胞核抗原(PCNA)，放射免疫法测定肝细胞DNA合成率。实验设150(H)、50(M)、15(L) mg·kg^-1·d^-1剂量组以及安慰剂(P)组和假手术对照组(F)。结果: 与P、F组相比，实验组大鼠肝细胞GPx和TR活性及PCNA指数和［³H］-TDR掺入水平均明显升高(P<0.05)。相关分析发现，GPx和TR活性与PCNA表达指数均有明显正相关(r²=0.77和0.87,P<0.05)，与［3H］-TDR掺入水平也具有高度正相关(r²=0.73和0.84,P<0.05)。结论：结果提示Se-SP对肝叶切除大鼠具有明显促肝细胞再生效应，这些硒酶活性上调可能在增强肝细胞再生增殖中起重要作用。     

层粘蛋白及其受体、增殖细胞核抗原与肝细胞癌肿瘤生物学特性的关系

目的:探讨层粘蛋白(LN)、层粘蛋白受体(LN-R)及增殖细胞核抗原(PCNA)与原发性肝细胞癌(HCC)恶性程度的关系。方法: 采用SABC法分别检测42例肝细胞癌及癌旁组织中LN、LN-R及PCNA的表达情况。 结果:LN、LN-R及PCNA在肝细胞癌中的表达均显著高于癌旁组织(P＜0.05),且与肝细胞癌Edmondson分级呈正相关;浸润型(IHCC)生长、有转移、肿瘤直径＞3 cm者LN、LN-R及PCNA的表达高,而膨胀型(EHCC)生长、无转移、肿瘤直径≤3 cm者多表达较低;在高增殖指数组LN、LN-R表达高于低增殖指数组(P＜0.05)。结论: 细胞内LN、LN-R及PCNA可作为评判肝细胞癌恶性程度及临床预后的良好指标。     

法尼酯X受体在肝硬化大鼠肝切除术后肝再生中的作用

目的 探讨法尼酯X受体(FXR)在肝硬化大鼠肝切除术后肝再生中的作用.方法 30只肝硬化大鼠肝切除术后按完全随机法分为对照组、胆酸组和消胆胺组饲养1周,每组10只.对照组给以标准饲料饲养,胆酸组给以含0.2%胆酸的饲料饲养,消胆胺组以含2%消胆胺的饲料饲养.1周后检测各组大鼠胆汁分泌速度及总胆汁酸(TBA)含量,检测肝功能、有丝分裂指数(MI)、增殖细胞核抗原(PCNA)标记指数和细胞核DNA含量,并应用RT-PCR和Western免疫印迹分别检测肝组织FXR、胆固醇7α羟化酶(CYP7A1)两者mRNA和蛋白的表达.结果 胆酸组胆汁分泌速度、TBA含量、血清白蛋白(ALB)含量、MI、PCNA标记指数、细胞核DNA含量均高于对照组(P<0.05),血清谷草转氨酶(AST)、谷丙转氨酶(ALT)均低于对照组(P<0.05).而消胆胺组胆汁分泌速度、胆汁TBA含量、血清ALB含量、MI、PCNA标记指数、细胞核DNA含量均低于对照组和胆酸组(P<0.05),血清谷草转氨酶(AST)、谷丙转氨酶(ALT)均高于对照组和胆酸组(P<0.05).胆酸组肝组织FXR mRNA和蛋白表达均高于对照组(mRNA:0.671±0.027比0.528±0.017,蛋白:0.702±0.039比0.566±0.020,P<0.05).肝组织CYP7A1mRNA和蛋白表达含量均低于对照组(mRNA:0.237±0.020比0.325±0.076,蛋白:0.264±0.015比0.325±0.084,P<0.05).而消胆胺组肝组织FXR mRNA(0.468±0.023)和蛋白表达(0.502±0.021)均低于对照组和胆酸组(P<0.05),肝组织CYP7A1 mRNA(0.411±0.021)和蛋白表达(0.476±0.018)则高于对照组和胆酸组(P<0.05).结论 胆汁酸与其受体FXR结合激活细胞内信号转导对肝硬化大鼠肝切除术后肝再生起重要作用,升高胆汁酸水平促进肝再生,降低胆汁酸水平则抑制肝再生.     

Rates of oxygen consumption and of anaerobic glycolysis in renal cortex and medulla of adult and new-born rats and guinea-pigs

1. Rates of oxygen uptake and of anaerobic glycolysis were estimated in slices from the renal cortex and medulla (a) of adult rats and guinea-pigs, (b) of new-born (1-, 5- and 21-day-old) rats and of guinea-pigs of 1, 12, 21, 24 and 120 hr age.2. In the adult rat, Q(O2) values for the cortex were 12.55 +/- 0.20 (22) and for the medulla: 8.56 +/- 0.17 (22) mul./hr.mg dry weight, while in the new-born rat (24 hr old) they were 10.99 +/- 0.46 (12) and 9.33 +/- 0.18 (9) mul./hr.mg dry weight respectively.3. Values for Q(CO2) (N2) (anaerobic glycolysis) in the 14 hr old new-born rat were in the renal cortex 9.65 +/- 0.35 (5) and in the medulla 7.39 +/- 0.43 (5) mul./hr.mg dry weight; while in the adult they were 2.25 +/- 0.08 (16) and 5.76 +/- 0.14 (16) mul./hr.mg dry weight, respectively.4. In the adult guinea-pig values for Q(CO2) (N2) were of the same order as in the adult rat, though the rate of O(2) uptake was for the cortex 8.12 +/- 0.22 (12) and for the medulla 5.02 +/- 0.23 (11) mul./hr.mg dry weight.5. Though the Q(O2) values in the renal cortex and medulla were smaller in the 1 hr old new-born guinea-pig, they were already increasing in the 12 hr old neonate.6. The results are discussed in the light of enzyme changes occurring during the process of maturation of the nephron as indicated by histochemical observations.     

Timing and intensity of early fevers and the development of allergies and asthma

BACKGROUND: Early childhood fevers appear to protect against later allergies and asthma. What is not known is the time in which fevers exert this effect and whether the degree of temperature increase is important. OBJECTIVE: We sought to examine the relationship between the time and degree of early fevers and later allergies and asthma. METHODS: Eight hundred thirty-five children from southeast Michigan were enrolled at birth. Clinic records from their first 2 years were abstracted for episodes of fever. At age 6 to 7 years, children underwent allergy testing. We examined fevers occurring within 6-month intervals in the first 2 years of life and outcomes at age 6 to 7 years. The primary outcome measures were allergic sensitization, asthma, asthma with allergic sensitization, and asthma without allergic sensitization. RESULTS: In the unadjusted analysis each episode of fever between 7 and 12 months of age was associated with a lower odds of allergic sensitization (odds ratio [OR], 0.71; 95% CI, 0.54-0.93) and asthma with allergic sensitization (OR, 0.43; 95% CI, 0.21-0.90) at age 6 to 7 years. Likewise, every 1 degrees C increase in the maximum temperature between 7 and 12 months was associated with a lower odds of allergic sensitization (OR, 0.77; 95% CI, 0.61-0.96) and asthma with allergic sensitization (OR, 0.62; 95% CI, 0.40-0.94). After adjusting for potential confounders, each episode of fever between 7 and 12 months was associated with a lower likelihood of asthma with allergic sensitization (adjusted OR, 0.33; 95% CI, 0.11-0.94) at age 6 to 7 years. CONCLUSIONS: Both the timing and intensity of childhood fevers appear to be important factors in the development of allergies and asthma.     

Modes of reinforcement and gender and culture of experimenter and subject: effects of "alien" and external reinforcement for Japanese and American men and women

In this study, we examined external and "alien" reinforcement (ER and AR. respectively) as a factor in social learning, and studied the combined effects of culture and reinforcement mode. A female (Experiment 1) and a male (Experiment 2) experimenters conducted experimental sessions. Both men and women, who grew up in the same culture as the experimenter, participated and performed the experimental task. A three-way interaction effect of experimenter gender, culture, and reinforcement mode was found on task performance. And the effect was more pronounced for a Japanese experimenter. A female and a male experimenters conducted Experiments 3 and 4, respectively; however participants this time were men and women who grew up in different cultures than the experimenter. Results indicated that the pattern of the subject gender and reinforcement mode interaction effect, when the experimenter was Japanese with American subjects, was exactly opposite to that when the experimenter was American. These experiments showed that AR was as effective for social learning as ER, and that the cultural backgrounds of experimenter and subject influenced AR and ER effectiveness.     

纳米水平分子成像与诊疗一体化研究进展与挑战

分子成像能以非侵入性的方式重现活体细胞的生理功能和生物学过程,提高疾病的早期和特异性诊断水平。纳米颗粒/材料具有物理性质可控性高、易于表面修饰、血液循环时间长和可功能化等优点,在疾病诊断与治疗中显示出巨大潜力。但如何阐明纳米材料多功能间的内在联系、解决其代谢及安全性等关键机制难题、实现纳米颗粒/材料多功能性到临床多功能...