磷酸化肌球蛋白轻链在大鼠慢性移植肾肾病早期病变中的作用及其机制

目的 探讨磷酸化肌球蛋白轻链(pMLC)在慢性移植肾肾病(CAN)中的作用与机制.方法 依照标准的CAN大鼠模型进行左肾原位移植,受者为LEW大鼠,供者为F344大鼠,另取雄性F344大鼠和LEW大鼠仅行单肾切除术,分别作为对照.分别于术后4、8及12周时,收集各组大鼠的24 h尿量,并检测其尿肌酐水平,计算肌酐清除率.然后取各组大鼠血液标本,测定血清肌酐水平.采用Banff分级标准评定各组肾小球肾病、肾小管萎缩、间质纤维化及血管内膜增厚程度.采用免疫组织化学法和采用蛋白质印迹法检测肾组织中磷酸化MLC(pMLC)和整合素连接激酶(ILK)的表达部位及表达水平.结果 移植组大鼠术后4周时肾间质可见单个核细胞浸润,12周时可见血管平滑肌细胞的移行与增殖.移植组大鼠各时相肾组织pMLC和ILK表达水平显著高于Lewis对照组及F344对照组,且随着移植时间的延长有逐渐增高趋势.大鼠移植肾组织中pMLC表达水平与24 h尿蛋白定量、血清肌酐水平、肾间质单个核细胞浸润、肾小动脉血管平滑肌细胞数量、Banff评分等呈显著正相关,相关系数(r)分别为0.273(P＜0.05)、0.434(P＜0.01)、0.525(P＜0.01)、0.676(P＜0.01)、0.570(P＜0.01),在移植后4周时,pMLC表达水平与肾小管间质ILK表达水平呈显著正相关,r=0.778(P＜0.01).结论 pMLC在慢性移植肾病早期病理变化中发挥重要作用,移植肾肾小管间质及肾小动脉中pMLC表达上调与ILK的作用机制相关.

Abstract：

Objective To investigate the role and mechanism of phosphate myosin light chain (pMLC) in the rat kidney of chronic allograft nephropathy (CAN) model. Methods The left donor kidneys from Fisher (F344) rats were orthotopically transplanted into Lewis recipients. Meanwhile, the F344 rats and LEW rats with resection of the right kidney served as control groups. Animals were harvested respectively at the 4th, 8th and 12th week after transplantation. The creatinine clearance rate (CCr) was calculated by urine creatinine of 24-h urine. Blood samples were collected from rats for determination of serum creatinine. The expression of pMLC was detected by using Western blotting and immunohistochernistry, and that of integrin-linked kinase (ILK) by using immunohistochemistry. Results Mononuclear cells infiltration of allografts was markedly aggravated as compared to the controls. Allografts got severe interstitial fibrosis and tubular atrophy at 12th week after transplantation. The expression of pMILC and ILK was up-regulated in the kidney of CAN rats after transplantation, and increased more significantly as the time went on. The expression of pMILC was significantly correlated with 24-h urine protein excretion (r= 0. 273, P＜0. 05), serum creatinine levels (r = 0. 434, P＜0. 01 ), the number of tubulointerstitial infiltrated mononuclear cells (r = 0. 525, P＜0. 01 ), the number of smooth muscle cells (SMC) in vascular wall (r= 0. 676, P＜0. 01 ) and the extent of interstitial fibrosis (r= 0. 570, P＜0. 01 ).There was a significantly positive correlation between ILK and pMLC in CAN rats at the 4th week after transplantation (r= 0. 778, P＜0. 01 ). Conclusion pMLC might play an key role in CAN, and the over-expression of ILK might be involve in the pathogenesis of CAN.     

膀胱移行细胞癌中p53、nm23和bcl-2的表达及临床意义

目的探讨膀胱移行细胞癌组织中p53、nm23和bcl-2的表达及临床意义。方法应用免疫组织化学ABC方法,检测60例膀胱移行细胞癌中p53、nm23和bcl-2的表达情况。结果60例膀胱移行细胞癌组织中,p53阳性率48．3％,nm23的阳I眭率50．0％,bcl-2的阳性率55．0％,p53、nm23和bcl-2的表达与膀胱移行细胞癌的病理分级和临床分期显著相关（P〈0．05）。结论p53、nm23和bcl-2的表达,提示膀胱移行细胞癌预后不良,可以作为评价膀胱癌患者预后的指标。     

肾结石合并肾盂鳞状细胞癌的早期诊断与治疗（附2例报告）

目的探讨。肾结石合并。肾盂鳞状细胞癌的早期诊断与治疗。方法报告肾结石合并肾盂鳞状细胞癌2例,结合相关文献探讨其早期诊断及治疗方法。结果2例均有典型肾绞痛病史及合并血尿,CT检查提示肾盂癌2例。2例术中经冰冻病理确诊后,行根治性患肾切除术,1例术后行全身化疗,5个月后死于全身衰竭,另一类术后随访18个月无复发。2例病理诊断均为鳞状细胞癌。结论肾结石合并肾盂鳞状细胞癌恶性程度高、预后差,临床表现缺乏特异性,易误诊和漏诊,提高对本病的认识,对可疑病例进行多项必要的辅助检查,有助于明确诊断。治疗以根治性患。肾切除为主。     

携带超抗原SEA基因的白蛋白纳米载体的构建及其治疗膀胱癌潜能研究

目的 构建一种非病毒载体基因投递系统-携带超抗原葡萄球菌肠毒素A(SEA)基因的白蛋白纳米载体,观察白蛋白纳米粒表征并探讨其潜在的靶向基因投递作用和强大的抗瘤机制.方法 采用去溶剂化法制备白蛋白纳米粒,平均粒径(253.1±11.9)nm,Zeta电位(-34.0±4.5)mV,PDI 0.43±0.04;抽提SEA基因质粒(pSEA)260/280:1.84±0.02,pSEA浓度:(85.54±1.43)mg/L;经生物活性及基因测序鉴定后耦合SEA基因质粒和白蛋白纳米载体,并观察耦合物的稳定性及白蛋白纳米载体对SEA基因的保护作用.结果 成功构建携带超抗原SFA基因的白蛋白纳米载体,平均粒径(118.9±4.8)nm,Zeta电位(-43.9±10.5)mV,PDI 0.19±0.02,基因转载率为(97.61±0.06)%,性质稳定、分散性较好,体外实验表明白蛋白纳米载体能保护SEA基因免受DNase Ⅰ的降解.结论 获得符合超抗原SEA基因转染要求的白蛋白纳米载体.

Abstract：

Objective To assess the characteristics of human serum albumin nanoparticles (HSA-NP) as a nonviral vector system for delivery staphylococcal enterotoxin A (SEA) gene and probe into its potential targeted antitumor mechanism.Methods HSA-NP and plasmid containing SEA gene (pSEA)encapsulated in HSA (pSEA-HSA-NP) were prepared by a desolvation-crosslinking method.HSA-NP had a mean size of (253.1 ± 11.9) nm,zeta potential of ( -34.0 ±4.5) mV,polydispersity index of 0.43 ±0.04.The superantigen SEA gene was extracted by the Endo-Free Plasmid Maxi Kit,260/280 of pSEA was 1.84 ±0.02 and the concentration of pSEA was ( 85.54 ± 1.43 ) mg/L.pSEA was verified by sequencing and biological activity survey,the stability of pSEA-HSA-NP was investigated by laying for 10 days at room temperature,and the size and zeta potential were remeasured and contrasted with the samples 10 days before.HSA-NP protecting pSEA from degradation of DNase I was detected by gel electrophoresis.Results Electrophoretic mobility analysis and fluorescent labeling revealed that pSEA-HSA-NP was successfully constructed.PSEA-HSA-NP had a mean size of ( 118.9 ±4.8) nm,zeta potential of ( -43.9 ± 10.5 ) mV,polydispersity index of 0.19 ±0.02 and pSEA encapsulation efficiency of (97.61 ±0.06)%.The characteristics of pSEA-HSA-NP solution laying for 10 days at room temperature indicated a better stability and polydispersity.HSA-NP stabilizing pSEA against DNase I in vitro was also testified by gel electrophoresis.Conclusion pPSEA-HSA-NP served the transfecting needs of superantigen SEA gene.     

后腹腔镜肾上腺切除术的临床分析

目的探讨后腹腔镜切除肾上腺病变的手术方法和效果。方法对23例肾上腺病变患者行后腹腔镜肾上腺切除术。经腰部置入3个Trocar,手指法建立后腹膜气腹,建立腹膜后操作通道。游离出病变肾上腺达中央静脉后以钛夹夹闭后切断,将切除的肾上腺置入标本袋后取出。结果本组除2例改为开放手术外均获得成功,手术时间55～300min,平均70min。术中出血10～100ml,平均30ml,所有病例均未输血。2例中转开放手术,均为腹膜破裂致后腹腔显露困难所致。无大出血、感染、腹腔脏器损伤等并发症发生。随访1～24个月,平均8个月,未发现肿瘤复发和转移。结论后腹腔镜肾上腺切除术是治疗肾上腺疾病的一种微创、安全、有效的手术方式。     

消毒供应室人员职业损伤因素分析与防护对策

目的分析消毒供应室人员存在的职业损伤并探讨其防护对策。方法回顾总结我院消毒供应室人员2008年1月至2010年9月期间发生的职业损伤。结果共发生145起职业损伤,其中物理性损害最为多见,占55．9％。化学性损害次之,占25．5％。其余分别为生物性损害（14．5％）,工种相关性损害（4．1咖。物理性损害中以皮肤划伤,割伤、刺伤最为多见,占75．3％（61/81）,其次为烧烫伤。化学性损害以消毒剂引起的皮肤损害曩为多见。结论针对各危害因素进行针对性防护,能有效减少职业危害,为工作人员身体健康提供保障。     

吸烟,高镉与尿石形成的关系

为进一步研究吸烟、高镉与尿石形成的关系,选用20只Wistar雄性大鼠,在膀胱内植入直径4mm的圆形金属锌片,后将动物随机分成实验组及对照组。两组均同时饮用1％乙二醇水,实验组每周吸烟5次,持续4周。实验前后测定大鼠24小时尿草酸、钙、磷、镉、β2-微球蛋白排泄量,用分级法评价结石形成及肾损伤程度,膀胱结石称其干重。结果显示吸烟组动物结石发生率高于对照组（P<0.01）,24小时尿草酸、钙、磷、镉和β2-微球蛋白排泄量显著高于实验前及对照组（P<0.01）,膀胱结石的体积、数目亦较对照组增加（P<0.01）。表明镉增高可促进泌尿系结石的形成。     

门诊癌症患者麻醉处方分析

目的分析中山大学附属第五医院门诊癌症患者麻醉药品使用情况。方法以限定日剂量（DDD）和药物利用指数（DUI）值为指标,对本院2006年1月至2007年1月4019张麻醉处方中的癌症患者所用3650张麻醉处方进行统计学分析。结果哌替啶注射液DUI值为1.25,盐酸吗啡缓释片为0.93,磷酸可待因片为0.88。结论本院癌症患者麻醉药品使用基本合理,但哌替啶注射液DUI值为1.25,与＂癌症三阶梯止痛疗法＂有一定的差距。     

男性膀胱癌患者血清性激素水平的检测

目的　探讨性激素在膀胱癌发病中的意义。方法 :采用放射免疫法检测 31例男性膀胱癌患者及 2 7例同年龄正常人对照组血清中的雌二醇 (EstradiolE2 )、孕酮 (ProgesteroneP)及睾酮 (TestosteroneT)水平。结果 :男性膀胱癌患者血清E2 、P、T分别为 44.98± 2 0 .96pg ml( x±s) ,0 .37± 0 .18ng ml( x±s) ,6 .2 5± 2 .37ng ml( x±s) ,而正常对照组E2 、P、T分别为 5 6 .0 1± 13.5 0pg ml( x±s) ,1.14± 0 .37ng ml( x±s) ,5 .15± 1.37ng ml( x±s) ,两者比较膀胱癌组T明显高于对照组 (P <0 .0 5 ) ;而E2 、P明显低于对照组 (P <0 .0 5 )。结论 :膀胱癌的发生可能与性激素代谢失衡有一定的关系