醛固酮及其受体拮抗剂对大鼠主动脉平滑肌TGF-β1表达的影响

目的 观察醛固酮及其受体拮抗剂对大鼠主动脉平滑肌转化生长因子-β1(TGF-β1)表达的影响,初步探讨原发性醛固酮增多症发生血管重构的可能机制.方法 采用皮下给药的方法建立醛固酮增多症模型,其中醛固酮组经微量渗透泵持续释放醛固酮(1 μg/h);拮抗组除给予等量醛固酮外,每日行螺内酯灌胃100 mg/(kg·d);对照组仅泵空白溶剂.尾套法检测大鼠血压,4周后处死所有大鼠,测定血钾、钠、醛固酮浓度及血浆肾素活性.分别采用RT-PCR、Western印迹检测主动脉平滑肌TGF-β1基因的表达.结果 ① 醛固酮组大鼠血压明显升高,血钾下降,血浆肾素活性降低,与对照组相比差异具统计学意义(P<0.01);② 醛固酮组主动脉平滑肌TGF-β1基因的表达水平明显高于对照组(P<0.01).螺内酯可抑制醛固酮的上述作用(P<0.01).结论 醛固酮及其受体拮抗剂螺内酯可能通过调节血管平滑肌TGF-β1的表达,从而影响血管重构的发生.     

The single needle method for urethrovesical anastomosis with strengthened posterior fixation during laparoscopic radical prostatectomy

The single needle method for urethrovesical anastomosis with strengthened posterior fixation during laparoscopic radical prostatectomy was explored. The method was initiated by performing a fixing suture with a knot at 4 o’clock of the posterior lip of bladder neck, and another suture at nearby position was performed to leave the knot outside. From 5 o’clock to 8 o’clock, sutures were performed every one o’clock to secure posterior approximation, then every two o’clock a suture. To avoid a loose anastomosis, lock sutures were performed every 3 sutures. The needle was always driven full-thickness outside-in in the bladder neck and inside-out on the urethra. After completing the full circumference, the needle was drawn near the 4 o’clock and tied at the tail end. Any leakage could be closed with additional interrupted sutures. The clinical data of 89 patients who underwent this method were retrospectively compared with those of 23 patients who underwent the single knot method. The results showed that the anastomosis, operative and catheterization time was 17.6±4.7 min, 134.0±10.7 min and 6.5±1.6 days respectively. There were 3 temporal urinary leakages identified in 89 cases requiring prolonged catheterization. No urinary leak and anastomotic stricture was confirmed, and 95.2% patients had total urinary control. It was concluded that this method was simple and safe for urethrovesical anastomosis.     

靶向沉默Notch1基因对人膀胱癌细胞生物学行为的影响

目的 探讨沉默Notch1基因对人膀胱癌细胞生物学行为的影响. 方法 将靶向Notch1的siRNA真核表达载体psiRNA Notch1-1转染膀胱癌细胞株T24和BIU-87,噻唑盐法、流式细胞术检测沉默Notch1后膀胱癌细胞生长、细胞周期和凋亡情况.RT-PCR和蛋白质印迹法检测转染前后Notch1基因mRNA和蛋白表达的变化. 结果 转染后72 h,T24和BIU-87细胞G0/G1期细胞比例明显增加,分别为(80.13±2.69)%和(69.44±2.41)%,与T24对照组(23.89±1.32)%和BIU-87对照组(24.63±1.68)%比较差异有统计学意义(P值均<0.05).转染后72 h,T24和BIU-87细胞凋亡率分别为(13.75±1.23)%和(8.72±1.01)%,与T24对照组(1.28±0.14)%和BIU-87对照组(1.01±0.27)%比较差异均有统计学意义(P值均<0.05).转染后24 h细胞生长明显受到抑制,该抑制作用持续到转染后96 h.转染后72 h,T24和BIU-87细胞中Notch1基因mRNA和蛋白表达明显下调(P<0.05). 结论 Notch1在膀胱癌细胞株中可能起致癌作用.沉默Notch1基因能够抑制膀胱癌细胞的增殖.     

Notch信号通路基因在膀胱癌中的表达及其与预后的关系

目的 探讨Notch信号通路在膀胱移行细胞癌中的表达及与预后间的关系.方法 收集膀胱移行细胞癌组织新鲜标本70例,10例正常膀胱黏膜作为对照组.采用免疫组织化学、逆转录-聚合酶链反应(RT-PCR)和Western blot检测Notch-1、Notch-2、Notch-3、Jagged-1和DLL-1的表达.对70例患者进行随访,结合各基因表达分析其与术后生存时间的关系.结果 在10例正常膀胱移行上皮中以上5种Notch信号通路基因均为深染.非浸润性膀胱癌组织中染色主要集中于细胞膜上,染色较淡,浸润性膀胱癌中染色部位分布于细胞质中,呈片状浓聚.RT-PCR和Western blot发现5种因子在三种组织中的表达差异有统计学意义(P＜0.05).非浸润性膀胱癌患者的术后无瘤生存时间与Notch-1和Jagged-1基因的表达状态相关(P＜0.05).结论 在正常膀胱黏膜上皮、非浸润性膀胱癌和浸润性膀胱癌中Notch信号通路基因的表达呈现3种模式.非浸润性膀胱癌中Notch-1和Jagged-1基因的低表达与患者预后不良相关.     

Aortic Cell Apoptosis in Rat Primary Aldosteronism Model

This study aimed to determine whether aldosterone could induce vascular cell apoptosis in vivo.Thirty-two male rats were randomly divided into 4 groups:vehicle(control),aldosterone,aldosterone plus eplerenone or hydralazine.They were then implanted with an osmotic mini-pump that infused either aldosterone or the vehicle.Systolic blood pressure(SBP) was measured weekly by the tail-cuff method.After 8 weeks,plasma aldosterone concentration(PAC) and renin activity(PRA) were determined by radioimmunoassay.Aorti...     

肾嗜酸细胞腺瘤国内外平均发病年龄比较及文献分析（附24例报告）

目的：掌握我国肾嗜酸细胞腺瘤患者的平均发病年龄,提高其诊治水平。方法：回顾性分析24例肾嗜酸细胞腺瘤患者临床资料：发病年龄22-84岁,其中〈40岁2例,40～49岁10例,50-59岁3例,60-69岁5例,70～79岁2例,80-89岁2例。同时复习相关文献,比较国内外该病的平均发病年龄。结果：本组24例平均发病年龄为54．5岁,术后病理检查证实均为肾嗜酸细胞腺瘤。统计国内肾嗜酸细胞腺瘤224例平均发病年龄为52岁,国外397例平均发病年龄为66岁,国内发病年龄较欧美早14年。结论：肾嗜酸细胞腺瘤平均发病年龄国内与国外报道有明显差别,应引起临床重视。     

Relationship between the Expression of RASSF1A Protein and Promoter Hypermethylation of RASSF1A Gene in Bladder Tumor

To investigate the relationship between the expression of RASSF1A protein and promoter hypermethylation of RASSF1A gene, RASSF1A protein expression was measured by Western blotting in 10 specimens of normal bladder tissues and 23 specimens of bladder transitional cell carcinoma （BTCC）. The promoter methylation in BTCC and normal bladder tissues was detected by methylation-specific PCR （MSP）. The results showed that the expression level of RASSF1A protein was significantly lower in BTCC tissues than that in normal bladder tissues. However, it was not correlated with its clinical stages and pathological grades. The frequency of promoter methylation of RASSF1A gene was higher in BTCC tissues than that in normal bladder tissues. In 14 patients with the aberrant promoter methylation, 13 showed loss or low expression of RASSF 1A protein. It is concluded that RASSF1A gene promoter methylation may contribute to the low level or loss of RASSF1A protein expression, the inactivation of RASSF1A gene and the genesis of BTCC. But, it may bear no correlation with its clinical stages and pathological grades.     

Expression of KAI1/CD82 and MRP-1/CD9 in Transitional Cell Carcinoma of Bladder

The expression of KAI1/CD82 and MRP-1/CD9 in transitional cell carcinoma of bladder (TCCB) and its clinical significance were investigated. Immunohistochemistry was used to detect KAI1/CD82 and MRP-1/CD9 protein expression in 52 TCCB specimens. Correlation between the expression of KAI1/CD82 and MRP-1/CD9 to clinicopathologic factors was statistically analyzed. The results showed that the positive rate of KAI1/CD82 and MRP-1/CD9 in TCCB was 50% and 61.5%, respectively. The MRP-1/CD9 and KAI1/CD82 expression was significantly associated with grade of TCCB (P<0.05), but no correlation was found between MRP-1/CD9 or KAI1/CD82 expres-sion and clinical stage of TCCB (P>0.05). The expression level of MRP-1/CD9 and KAI1/CD82 in recurrent TCCB samples was lower than that in non-recurrent samples (P<0.05). Meanwhile, the correlation between the KAI1/CD82 expression and MRP-1/CD9 expression was statistically signifi-cant (r=0.316, P<0.05). It was concluded that KAI1/CD82 and MRP-1/CD9 expression may be im-portant prognostic indicators and potentially useful for assessing the biological behavior of TCCB.     

MGMT基因启动子甲基化与膀胱尿路上皮癌生物学行为关系的研究

目的 检测膀胱尿路上皮癌组织中O6-甲基鸟嘌呤-DNA甲基转移酶(MGMT)基因启动子甲基化状态,探讨MGMT甲基化与其蛋白表达水平以及肿瘤生物学行为之间的关系.方法 应用免疫组织化学SP法和甲基化特异性PCR(MSP)分别检测60例膀胱尿路上皮癌及15例正常膀胱黏膜组织中MGMT蛋白表达情况和MGMT基因启动子甲基化状态.结果 膀胱癌组织中MGMT蛋白阳性表达率为35.0 %(21/60),低于正常膀胱组织(86.7 %,13/15,P<0.01).膀胱癌组织中MGMT甲基化阳性率为45.0 %(27/60),明显高于正常膀胱组织(0.0 %,0/15,P<0.01);MGMT启动子甲基化与其蛋白表达呈负相关(r = -0.453,P<0.01);并且高级别膀胱癌中MGMT甲基化阳性率(70.6 %,12/17)要比低级别膀胱癌高(34.9 %,15/43),(P<0.05),而MGMT甲基化与膀胱癌临床分期无明显关系.结论 MGMT启动子甲基化可能参与了膀胱尿路上皮癌的发生和肿瘤分化,MGMT启动子甲基化有望成为预判膀胱癌预后的重要标记.     

内源性硫化氢及其合成酶在膀胱癌恶性进展中的表达

目的探讨内源性硫化氢(H2S)及其合成酶胱硫醚-β-合成酶(CBS)、胱硫醚-γ-裂解酶(CSE)和3-巯基丙酮酸硫转移酶(MPST)在正常膀胱组织以及膀胱癌中的表达。方法选取因膀胱癌而行膀胱肿瘤电切术或膀胱癌根治术的肿瘤组织标本76例,同时收集15例正常膀胱组织作对照组。通过免疫组化和Western blot研究CBS、CSE和MPST在正常膀胱组织以及不同病理分期/分级膀胱癌组织中的表达情况;通过敏感硫电极法检测H2S在正常膀胱组织和膀胱癌中的含量。分别比较H2S及其合成酶在各组之间的差异,初步探讨H2S及其合成酶与膀胱癌恶性进展之间的关系。结果免疫组化结果显示CBS(79.1%)、CSE(81.3%)和MPST(84.6%)在膀胱癌黏膜和肌层组织中均有表达。CBS(P=0.021和P<0.001)和MPST(P=0.015和P=0.045)的表达在膀胱癌不同分期和分级之间有统计学差异。在所有检测的标本中,肌层浸润性膀胱癌中的H2S含量最高[(52.6±2.91)nmol·mg-1·min-1],各组之间有统计学差异。结论内源性H2S及其合成酶CBS、CSE和MPST在膀胱癌组织中的表达明显高于正常膀胱组织,且其表达在膀胱癌不同分期和分级之间有统计学差异。