肾透明细胞癌miRNA-34a、Notch1和Hes1的表达及意义

目的观察miRNA-34a、Notch1和Hes1在肾透明细胞癌组织的表达，探讨其与肾癌发生发展的关系及可能的临床意义。方法运用实时荧光定量PCR方法检测miRNA-34a、Notch1和Hes1在肾透明细胞癌组织及其相应的癌旁正常组织的表达，并分析其与临床病理特征的关系。结果与正常组织相比，miRNA．34a在肾癌组织中高表达（P〈0．05），Notch1、Hes1在肾癌组织中低表达（P〈0．05）；miRNA-34a随着肾癌病理分级的增高而增高，Notch1的表达则随着肾癌病理分级的增高而降低（P均〈0．05）。结论高表达的miRNA-34a通过下调其靶基因Notch1参与肾透明细胞癌的发生发展，此有望成为肾癌诊断的分子标志物及潜在的治疗靶点。     

肾缺血对缺氧诱导microRNAs及VEGF-NOTCH信号分子的影响

Objective To investigate the expression changes of microRNAs and VEGF-NOTCH in renal ischemic injury in mice, and to explore the potential mechanism associated with renal angiogenesis.Method Male Balb/c mice were subjected to a standard renal ischemia to induce acute kidney injury (AKI) after 45 min of bilateral renal artery clamping. Following 4 h, 24 h of reperfusion or sham operation, kindey tissues were collected and subjected to detect the expression changes of microRNAs which relatived with angiogenesis and VEGF, Flk-1, Notch1 mRNA by Quantitative Real-time RT-PCR. Flk-1 protein was detected by Western blotting analysis at 24 h and 72 h following Ischemia/Reperfusion(I/R) injury. The expression of CD31 was examined in tissue sections by immunohistochemistry staining, and the microvessels in ischemic region of each group were counted. Results miRNA-210 and miRNA-92a expression increased significantly, with prominent changes at 4 h and 24 h after reperfusion( P ＜ 0.05 ). VEGF and Flk-1 mRNA expression and Flk-1 protein were increased in renal I/R compared with control group respectively (P＜0.05 ).Immunohistochemistry staining results of CD31 showed a significant increase of microvessels in renal ischemic region. Conclusion This study first reported the changes in miRNAs expression in response to kidney I/R in mouse. our results implied that miRNAs may be involved in targeting VEGF-Notch pathway signaling to regulate angiogenesis after renal I/R injury. It provided novel insights into the angiogenesis mechanism of renal ischemic injury.     

骨髓干细胞移植对急性缺血性肾损伤大鼠肾小管细胞活性的影响

目的观察和分析骨髓干细胞(BMSCs)移植对大鼠急性缺血性肾损伤后肾小管上皮细胞坏死、变性及增殖的影响。方法 Percoll密度梯度离心法分离获取BMSCs。制作大鼠急性缺血性肾损伤模型,通过下腔静脉进行BMSCs移植。分别于缺血再灌注后24h、48h、7d、14d获取肾脏标本,HE染色作肾组织学观察,免疫组织化学染色检测增殖细胞核抗原(PCNA)的表达。结果缺血再灌注后24h、48h,BMSCs移植组肾小管上皮细胞未见坏死及明显变性,而对照组细胞坏死及变性明显;缺血再灌注后7d、14d,BMSCs移植组和对照组肾小管上皮均无细胞坏死,但对照组可见部分细胞胞浆肿胀及间质内少许红细胞;BMSCs移植组较对照组肾小管上皮PCNA阳性细胞数增多,组间差异具统计学意义。结论 BMSCs移植可减少急性缺血性肾损伤的肾小管上皮细胞变性、坏死,而促进肾小管上皮细胞的增殖。     

腹腔镜膀胱阴道瘘修补术

目的 探寻治疗膀胱阴道瘘（VVF）微创、有效和安全的新方法。方法 2010年10月至2012年4月，应用腹腔镜膀胱阴道瘘修补术对15例VVF患者行VVF修补。结果 15例腹腔镜VVF修补术均顺利完成，手术时间为90～210 min，术中出血量为20～60 mL，随访时间为10～28个月，均未复发。结论 腹腔镜膀胱阴道修补术安全、有效且创伤小，长期疗效有待更多的临床资料验证。     

p16和p53抑癌基因在肾母细胞瘤中的表达及与预后关系

目的 探讨p16、p53抑癌基因在肾母细胞瘤中的表达及与临床预后的关系。方法 应用免疫组织化学方法S-P和ABC法检测p16和p53基因的表达。结果 20例肾母细胞瘤标本p16蛋白染色阳性7例，阳性率为35％；其中上皮型10例，阳性4例(占40％)；间叶型5例，阳性1例(占20％)；胚胎型3例，阳性1例(占33％)；混合型2例，阳性l例(占50％)。p53蛋白阳性染色5例，阳性率为25％。其中上皮型及混合型12例中，p53阳性2例(占16．7％)；而间叶型及胚胎型8例中，阳性3例(占37．5％)。结论对肾母细胞瘤患同时进行p16和p53检测更能准确地判断其病理性质和预后。     

尿道修复材料的研究及应用进展

尿道修复材料是尿道修复重建成败的关键,将尿道的修复材料分为自体组织材料、人工合成材料及组织工程化材料,从实验室和临床2方面对尿道修复材料的研究及应用进展进行综述.     

尿道修复材料的研究及应用进展

尿道修复材料是尿道修复重建成败的关键,将尿道的修复材料分为自体组织材料、人工合成材料及组织工程化材料,从实验室和临床2方面对尿道修复材料的研究及应用进展进行综述.     

经会阴途径后尿道端端吻合术治疗创伤性后尿道狭窄37例

目的探讨经会阴途径后尿道端端吻合术治疗创伤性后尿道狭窄的临床疗效。方法 37例创伤性后尿道狭窄患者术前均常規行膀胱尿道造影,复杂病例加行尿道B超、多层CT尿道重建(CTU)、磁共振尿道成像(MRU)及尿道镜或软性膀胱镜检查。37例患者均行经会阴途径后尿道端端吻合术,其中单纯尿道端端吻合术11例,阴茎海绵体中隔切开+后尿道端端吻合术23例,阴茎海绵体中隔切开+耻骨下缘楔形切除+后尿道端端吻合术3例。结果 37例患者术后随访3~20个月,平均12.3个月,术后最大尿流率为13.3~51.2 mL.s-1,平均(21.81±8.04)mL.s-1。手术总的成功率为86.5%(32/37),其中经会阴单纯尿道端端吻合术的成功率为90.9%(10/11),阴茎海绵体中隔切开+后尿道端端吻合术的成功率为86.9%(20/23),阴茎海绵体中隔切开+耻骨下缘楔形切除+后尿道端端吻合术的成功率为66.7%(2/3);手术不成功5例(13.5%)。结论经会阴途径后尿道端端吻合术可有效治疗创伤性后尿道狭窄。     

后肾细胞微环境诱导胚胎干细胞向肾系细胞分化的实验研究

目的: 探讨胚胎后肾细胞微环境诱导胚胎干细胞(ESCs)分化为肾系细胞的作用。方法: 小鼠D3系胚胎干细胞通过悬滴法制备拟胚体(EBs),将EBs细胞与取自孕12.5 d小鼠胚胎的后肾细胞通过间接共培养以诱导其分化,即为共培养诱导组,设EBs细胞自然分化为对照组;采用免疫荧光染色法检测共培养第3、5、7 d后的EBs细胞Pax2、WT-1蛋白表达情况,通过逆转录PCR法检测诱导3 d后的EBs细胞Pax2、WT-1、Lim1、Sall1、Emx2、GDNF、Wnt4、BMP7、Nephl、Nephrin、KSP和CD24 mRNA的表达情况。结果: EBs细胞在诱导的第3 d即出现了全部肾发育相关基因的表达,其中共培养组Pax2、WT-1、Emx2、GDNF、Nephl、Nephrin、KSP和CD24的表达强于对照组;免疫荧光结果显示在诱导3 d后的EBs细胞中可观察到Pax2阳性细胞,阳性细胞数在共培养第5、7 d增多;诱导5 d后可观察到WT-1阳性细胞,对照组未见Pax2或WT-1蛋白阳性细胞出现。结论: 后肾细胞微环境可促进ESCs分化为肾系细胞。     

肾缺血对缺氧诱导microRNAs及VEGF-NOTCH信号分子的影响

目的 观察肾缺血性损伤后,缺氧诱导microRNAs及VEGF-NOTCH信号分子的表达变化,探讨肾缺血损伤后血管新生的可能调控途径.方法 选择雄性Balb/c小鼠20只,采用夹闭双侧肾蒂方法制备急性缺血肾损伤模型,假手术组设为对照组.通过观察缺血恢复后24 h小鼠肾功能及肾组织病理学改变确定模型成功.实时定量RT-PCR检测缺血恢复后4 h,24 h时缺氧诱导miRNA-210,miRNA-92a,VEGF,Flk-1及Notch1 mRNA表达水平;Western-blot检测缺血恢复后24 h,72 h时Flk-1蛋白的变化;免疫组织化学染色检测CD31表达,判断缺血组织微血管内皮细胞增生状况.结果 肾缺血恢复24 h小鼠肌酐、尿素氮明显升高(P＜0.05),光镜下观察大量小管上皮细胞肿胀、空泡变性坏死,肾小管管腔扩张,见上皮细胞碎片和管型,表明成功建立肾缺血损伤模型.肾缺血恢复4 h,24 h后,与正常组比较,肾组织miRNA-210上调倍数分别为(2.02±0.29),(5.58±0.16);miRNA-92a的表达上调倍数分别为(3.23±0.74),(1.53±0.33),P＜0.05;VEGF,Flk-1及Notch1 mRNA表达水平均升高(P＜0.05).缺血恢复后24 h,72 h时,Flk-1蛋白水平显著升高(P＜0.05),肾组织微血管密度明显增加.结论 肾缺血性损伤后可出现代偿性血管新生,且缺氧诱导miRNA如miRNA-92a,miRNA-210表达上调,与血管新生相关的信号分子VEGF及Notch1表达均升高,提示miRNA/VEGF-Notch1可能是肾缺血性损伤后血管新生的主要调控通路,从而为探讨缺血性损伤后血管新生的机制提供了依据.

Abstract：

Objective To investigate the expression changes of microRNAs and VEGF-NOTCH in renal ischemic injury in mice, and to explore the potential mechanism associated with renal angiogenesis.Method Male Balb/c mice were subjected to a standard renal ischemia to induce acute kidney injury (AKI) after 45 min of bilateral renal artery clamping. Following 4 h, 24 h of reperfusion or sham operation, kindey tissues were collected and subjected to detect the expression changes of microRNAs which relatived with angiogenesis and VEGF, Flk-1, Notch1 mRNA by Quantitative Real-time RT-PCR. Flk-1 protein was detected by Western blotting analysis at 24 h and 72 h following Ischemia/Reperfusion(I/R) injury. The expression of CD31 was examined in tissue sections by immunohistochemistry staining, and the microvessels in ischemic region of each group were counted. Results miRNA-210 and miRNA-92a expression increased significantly, with prominent changes at 4 h and 24 h after reperfusion( P ＜ 0.05 ). VEGF and Flk-1 mRNA expression and Flk-1 protein were increased in renal I/R compared with control group respectively (P＜0.05 ).Immunohistochemistry staining results of CD31 showed a significant increase of microvessels in renal ischemic region. Conclusion This study first reported the changes in miRNAs expression in response to kidney I/R in mouse. our results implied that miRNAs may be involved in targeting VEGF-Notch pathway signaling to regulate angiogenesis after renal I/R injury. It provided novel insights into the angiogenesis mechanism of renal ischemic injury.