NF-κB在肝细胞癌组织中的表达及意义

目的：探讨NF-κB基因在肝细胞癌组织中的表达及生物学意义。方法：采用逆转录聚合酶链反应(RT-PCR)、Western-blot免疫组织化学技术检测32例肝癌组织及癌旁肝组织中的NF-κB基因的mRNA和蛋白表达水平及蛋白定位。结果：NF-κB基因在肝癌组织中的表达较癌旁肝组织显著性增高(P<0.05)。免疫组化结果显示，NF-κB蛋白在肝癌细胞胞浆和胞核中均有表达，而在癌旁肝细胞中仅在胞浆中有表达。结论：NF-κB基因在肝癌组织中呈显著高表达，提示其可能参与了肝癌的发生发展。NF-κB表达定位在癌细胞和癌旁肝细胞中的差异，提示NF-κB活化后，进入胞核，通过调节下游基因的转录，促进肝癌的发生。     

survivin与nmp22对诊断尿路上皮肿瘤价值的比较

目的 :评价尿脱落细胞中survivin的表达和尿nmp2 2的表达对诊断尿路上皮肿瘤的价值。 方法 :对4 8例尿路上皮肿瘤患者 ,在行膀胱镜检查或手术前留新鲜尿液分别行尿脱落细胞survivin、尿nmp2 2和尿脱落细胞检查 ,并分别比较各方法的敏感性、特异性。结果 :尿路上皮肿瘤患者尿脱落细胞涂片中survivin表达 4 8例中4 6例阳性 ,尿NMP2 2有 38例阳性 ,而尿脱落细胞学仅 15例阳性 ,survivin的敏感性高于nmp2 2及脱落细胞学 (P <0 .0 5及P <0 .0 1)。三者的特异性分别为 10 0 %、90 %和 10 0 % ,差异无统计学意义。结论 :检测尿脱落细胞中survivin的表达方法简单无创敏感性、特异性高对诊断尿路上皮肿瘤的价值优于尿nmp2 2。     

JC病毒样颗粒可直接转运进入细胞核

目的探讨JC病毒(JCV)病毒样颗粒(VLP)是否可以直接转运进入细胞核。方法心用JCV主要外壳蛋白VP1体外表达、重组VIP，在其表面标记异硫氰基荧光素(FTTC)，同时在其内部包裹荧光染料Cy3，感染培养的HeLa细胞和SVG细胞，荧光显微镜观察VLP入核转运。结果HeLa和SVG细胞感染包裹Cy3的FTTC-VLP时，FTTC与Cy3同时出现于细胞核内相同部位；而感染FTTC—VP1与Cy3混合物时，FTTC虽可在细胞核内检测到，但Cy3信号几乎消失。包裹Cy3的VIP用SDSPAGE展开，荧光显像后行考马斯亮蓝(CBB)染色，发现Cy3和VLP移行至不同部位，证明Cy3不能与VP1结合，提示VLP以完整的颗粒形式转运进入细胞核。应用包裹外源性DNA的VLP感染培养的HeLa和SVG细胞，发现包裹的DNA在细胞浆和细胞核内均可检测到，提示JCV入核过程与VIP相同。结论VLP可以不经裂解直接转运进入细胞核，JCV入核转运可能与VLP相同。     

2型糖尿病肾病患者血清中TNF-α、NO与ET的水平变化及其临床意义

目的：探讨2型糖尿病肾病患者血清肿瘤坏死因子仅（TNF-α）、一氧化氮（NO）和内皮素（ET）的水平变化及其临床意义。方法：用ELISA法检测91例2型糖尿病患者血清TNF-α水平，NO与ET的水平分别用硝酸还原酶法和放射免疫法测定。结果：2型糖尿病肾病各组患者TNF-α和ET水平较对照组明显升高，其中ODN组最高（P〈0．01）。而2型糖尿病肾病各组患者血清NO水平较对照组明显减少（P〈0．01）。显性糖尿病肾病患者血清TNF-α和ET呈正相关；血清TNF-α和NO、ET和NO均成负相关。结论：TNF-α、NO与ET可能参与2型糖尿病肾病的发病及病程变化过程，检测患者TNF-α、NO与ET水平可作为判断预后、指导治疗的指标。     

糖尿病大鼠骨折后骨形态发生蛋白-2、胰岛素样生长因子-1的变化

目的 观察糖尿病大鼠骨折后骨形态发生蛋白-2(BMP-2)、胰岛素样生长因子-1(IGF-1)的变化,探讨糖尿病对骨折愈合的影响.方法 70只大鼠随机分为对照组和糖尿病组,均造成左侧胫骨骨折.定期摄X线片,取骨痂HE染色,免疫组化检测骨痂BMP-2、IGF-1,ELISA法检测血清BMP-2、IGF-1.结果 2周BMP-2灰度值:实验组为149±8,对照组为107±7(P＜0.01);2周IGF-1灰度值:实验组为137±9,对照组为103±8(P＜0.01).2周血清BMP-2含量:实验组为(3.45±0.12) ng/ml,对照组为(5.60±0.11) ng/ml(P＜0.01);2周IGF-1含量:实验组为(5.89±0.12) ng/ml,对照组为(8.36±0.11) ng/ml(P＜0.01).结论 糖尿病大鼠骨折后血清及骨痂中BMP-2、IGF-1减少是导致糖尿病大鼠骨折愈合差的原因之一;BMP-2与IGF-1可能存在相互作用.     

The effect of single and repeated UVB radiation on rabbit cornea

Background Cumulative effect of ultraviolet radiation (UVR) is an important aspect of UV corneal damage. The purpose of this study was to apply high resolution magic angle spinning proton nuclear magnetic resonance (HR-MAS ¹H NMR) spectroscopy to evaluate the effect of single and repeated UV radiation exposure of the same overall dose on the rabbit cornea.Methods Corneal surfaces of 24 normal rabbit eyes were examined for the effects of UVB exposure (312 nm). In the first group (UVB1), animals were irradiated with a single dose (3.12 J/cm²; 21 min) of UVB radiation. The animals in the second group (UVB2) were irradiated three times for 7 min every other day (dose of 1.04 J/cm²; days 1, 3, 5) to give the same overall dose (3.12 J/cm²). The third group served as an untreated control group. One day after the last irradiation, the animals were sacrificed, and the corneas were removed and frozen. HR-MAS ¹H NMR spectra from intact corneas were obtained. Special grouping patterns among the tissue samples and the relative percentage changes in particular metabolite concentrations were evaluated using modern statistical methods (multivariate analysis, one-way ANOVA).Results The metabolic profile of both groups of UVB-irradiated samples was significantly different from the control corneas. Substantial decreases in taurine, hypo-taurine and choline-derivatives concentrations and substantial elevation in glucose and betaine levels were observed following the UVR exposure. There was no significant difference between the effect of a single and repeated UVB irradiation of the same overall dose.Conclusions For the first time, the effects of single and repeated UVR doses on the metabolic profile of the rabbit cornea were analysed and compared. The combination of HR-MAS ¹H NMR spectroscopy and modern statistical methods (multivariate analysis, one-way ANOVA) proved suitable to assess the overall view of the metabolic alterations in the rabbit corneal tissue following UVB radiation exposure.     

ApoE-/-小鼠肾动脉粥样硬化斑块破裂的肾损害

目的 探讨ApoE^-/-小鼠肾动脉粥样硬化斑块破裂对下游肾脏的损伤机制。 方法 采用ApoE^-/-小鼠建立粥样硬化性肾动脉狭窄(ARAS)动物模型。选择肾动脉狭窄程度 <50％的ApoE^-/-小鼠，按斑块分为破裂组和未破裂组；选择同条件喂养的C57BL/6J 野生型小鼠为对照组。常规检测Scr及尿 N-乙酰-β-氨基葡萄糖苷酶(NAG)活性；Western印迹检测细胞核中核转录因子κBp65（NF-κBp65）、细胞间黏附分子1（ICAM-1）及P-选择素（P-sel）表达； RT-PCR检测白细胞介素6 （IL-6）mRNA表达；免疫组化染色检测巨噬细胞浸润情况。 结果破裂组Scr和尿NAG活性明显升高（均P < 0.01）；肾组织出现病理改变，肾间质中巨噬细胞浸润增加（P < 0.05）；细胞核中NF-κBp65表达增加（P < 0.05）；ICAM-1、P-sel、IL-6 mRNA表达增加（P < 0.05）。未破裂组上述指标与对照组比较，差异无统计学意义（P > 0.05）；肾脏未见明显病理改变。 结论 肾动脉粥样硬化斑块破裂可引起肾脏病理改变和肾功能受损；在粥样硬化性肾动脉狭窄的肾损害机制中，炎性反应是重要因素之一。     

吡咯烷二硫氨基甲酸酯对大鼠重症急性胰腺炎的影响

目的　探讨核转录因子 (NF κB)抑制剂吡咯烷二硫氨基甲酸酯 (PDTC)对大鼠重症急性胰腺炎 (SAP)的影响。方法　将 3 6只Wistar大鼠随机分组 :假手术组 (n =6) ,假手术 静脉注射组 (n =6) ,SAP组 (n =12 )和试验组 (n =12 )。各组于造模 6h后 ,测定血清淀粉酶及脂肪酶含量 ,取胰腺组织进行病理学评分 ,采用免疫组织化学法检测NF κB激活水平及胰腺细胞凋亡情况。结果　SAP组和试验组的胰腺细胞内NF κB呈激活状态 ,存在胰腺细胞凋亡 ,与前两组差异有显著性 (P <0 .0 1) ,试验组大鼠的胰腺组织病理学评分、血清淀粉酶及脂肪酶含量、NF κB激活及细胞凋亡水平与SAP组差异存在显著性 (P <0 .0 5 )。结论　在SAP发病机制中 ,NF κB是多种炎症介质的始动因子 ,PDTC可以有效抑制胰腺细胞中NF κB的激活 ,促进胰腺细胞凋亡 ,减少胰酶释放 ,减轻胰腺组织的病理损害。     

The estimation of mean nuclear volume in the diagnosis of breast carcinoma.

Sixty-two cases of breast pathology were randomly selected from the files of the Dunedin Public Hospital for evaluation of mean epithelial nuclear volume. The cases were comprised of both benign and malignant ductal epithelial disease, diagnosed in cytological smears or in histological sections. Nuclear volume in histological preparations was estimated by the stereological technique of point-line intercept measurements to derive volume-weighted mean volumes (vV). An index of the nuclear volume (Vi) of cytology smears was calculated from measurements of nuclear areas by either image analysis Vi(e) or point-line intercepts Vi(p). By all methods of analysis a clear distinction of nuclear volume was found between the benign [means for the cytology were 148 microns 3 (Vi(e)) or 246 microns 3 (Vi(p)), and 203 microns 3 (Vv) for the histology specimens] and malignant diseased cases [means for cytology: 524 microns 3 (Vi(e)) or 886 microns 3 (Vi(p)), and 587 microns 3 (vV) for the histology specimens].     

NF-κB对尿白蛋白诱导肾小管上皮细胞骨桥蛋白转录的影响

目的研究NF-κB对尿白蛋白诱导肾小管上皮细胞骨桥蛋白转录的影响。方法去脂的小牛血清白蛋白(BSA)20mg/ml分别加入到体外培养的肾小管上皮细胞和加入NF-κB抑制剂(PDTC)共同培养半小时的肾小管上皮细胞内,EMSA、RT-PCR检测不同时限NF-κB活性、骨桥蛋白mRNA表达情况。结果去脂的小牛血清白蛋白可刺激肾小管上皮细胞内NF-κB活性增加,骨桥蛋白mRNA表达上调,而提前加入NF-κB抑制剂(PDTC)共同培养半小时的肾小管上皮细胞内NF-κB活性、骨桥蛋白mRNA表达明显下降。结论尿白蛋白诱导小管上皮细胞损伤的机制与诱导肾小管上皮细胞内NF-κB活性、骨桥蛋白mRNA增加有关,而且骨桥蛋白转录由NF-κB调控。