经肾窦内肾盂、肾后下段间切开加输尿管镜下钬激光碎石术治疗肾巨大鹿角形结石的临床应用（附36例报告）

目的:探讨肾巨大鹿角形结石的有效治疗方法.方法:分析总结36例巨大鹿角形肾结石患者经肾窦内肾孟、肾后下段间切开取石加输尿管镜下钬激光碎石的临床资料.结果:本组36例,均成功取石,平均手术时间120 min,结石一次性取净31例(86%),残留结石5例(14%),直径＜3 mm.其中3例自行排出,余2例经药物排石治疗后排出.2例术中出血量约250 mL,输血200～400 mL.术中无需阻断肾蒂,术前肾功能不全者术后均逐渐恢复正常.结论:经肾窦内肾孟、肾后下段间切开加输尿管镜下钬激光碎石术治疗.肾巨大鹿角形结石,手术视野清晰,能取净结石,可减少手术对肾实质的损伤,最大限度地保存患肾并可改善肾功能.手术安全有效,操作简单,损伤小,并发症少,残石率低,效果满意.     

抗凋亡因子XIAP和PED/PEA-15在前列腺癌(PC-3)中的表达及对细胞凋亡的影响

目的 检测抗凋亡因子XIAP与PED／PEA-15在前列腺癌细胞（PC-3）中的表达，探讨二者对前列腺癌细胞凋亡的影响。方法 应用半定量RT—PCR法检测前列腺癌细胞（PC-3）中PED／PEA-15和XIAP的表达。设计并构建PED／PEA-15和XIAP特异的siRNA载体，以脂质体法转染二者的siRNA载体至前列腺癌细胞（PC-3）中，半定量RT-PCR法检测特异siRNA载体对PED／PEA-15和XIAP转录的影响；光镜观察细胞形态改变；流式细胞法检测细胞凋亡的变化。结果 半定量RT-PCR显示PED／PEA-15和XIAP均在前列腺癌细胞（PC-3）中高表达。酶切和DNA测序证实XIAP和PED／PEA-15siRNA载体构建成功。共转染XIAP和PED／PEA-15siRNA载体入PC-3细胞，可导致XIAP和PED／PEA-15的转录抑制，并增加PC-3细胞对阿霉素的敏感性，凋亡明显增加，处理组凋亡率为79％，对照组为46％，两组差异有统计学意义（P〈0．05）。结论 PED／PEA-15和XIAP在前列腺癌的凋亡中可起重要作用。     

靶向人肝素酶短发卡状RNA表达载体的构建及其沉默作用

目的 构建靶向人肝素酶(HPSE)的短发卡状RNA(shRNA)表达载体,探讨其基因沉默作用.方法 根据人肝素酶mRNA序列设计shRNA,合成两条互补的寡核苷酸链,退火后连接入pGenesil-1载体,转化扩增后进行测序鉴定;重组质粒在阳离子脂质体Lipofectamine 2000的介导下,瞬时转染人胃癌细胞株SGC-7901、人前列腺癌细胞株PC-3、人膀胱癌细胞株EJ,每种细胞分别设空白对照组、空载体pGenesil-Negative转染组、pGenesil-HPSE shRNA转染组3组,逆转录-聚合酶链反应(RT-PCR)及Western blot法检测各种细胞中肝素酶基因表达水平,黏附实验和transwell小室实验检测癌细胞游走、侵袭能力.结果 所构建的shRNA载体插入基因片段与设计序列完全匹配,载体构建成功;重组质粒转染SGC-7901、PC-3、EJ细胞后,肝素酶mRNA表达分别下调78.6%(P<0.01)、82.6%(P<0.01)、81.9%(P<0.01),肝素酶蛋白表达分别下调76.4%(P<0.01)、85.9%(P<0.01)、83.3%(P<0.01),细胞黏附能力分别下降37.7%(P<0.01)、44.6%(P<0.01)、43.8%(P<0.01),细胞侵袭能力分别下降66.8%(P<0.01)、76.6%(P<0.01)、80.5%(P<0.01).结论 成功构建了靶向人肝素酶的shRNA表达载体,沉默肝素酶基因表达后,能抑制多种癌细胞的游走和侵袭能力.     

Effect of Smac on TRAIL-induced apoptosis of prostate cancer cell line PC-3 and the molecular mechanism

The effect of Smac gene on the TRAIL-induced apoptosis of the prostate cancer cell line PC-3 and the molecular mechanism were investigated. The Smac gene was transfected into PC-3 cells under the induction of liposome. The intrinsic Smac gene expression was detected by Western blotting. After treatment with TRAIL as an apoptosis inducer, in vitro cell growth activity was as-sayed by MTT colorimetry. The apoptosis rate of PC-3 cells was determined by annexin Ⅴ-FITC and propidium iodide staining flow cytometry. The expression of cellular XIAP and caspase-3 genes was examined by Western blotting. Smac-transfected cells (PC-3/Smac group) had significantly in-creased Smac protein level as compared with PC-3 controls (P<0.01). After induction with 100-200 ng/mL TRAIL for 12-36 h, cellular proliferation rate in PC-3/Smac group was significantly lower than in PC-3 controls (P<0.05). After induction with 100 ng/mL TRAIL for 24 h, the apoptosis rate in PC-3/Smac group was significantly enhanced as compared with that of PC-3 controls (P<0.05). Ac-cordingly, the XIAP expression level was down-regulated significantly (P<0.05) and caspase-3 sub-unit P20 was up-regulated significantly (P<0.05). It is suggested that the over-expression of cellular Smac can inhibit inhibitor of apoptosis proteins (IAPs), enhance caspases activity and the apoptosis rate of PC-3 cells induced by TRAIL, which may provide a useful experimental basis for prostate cancer therapy.     

精道造影在无精子症诊断中的临床应用57例分析

2001年2月至2008年4月作者对57例疑诊为梗阻性无精子症患者施行精道造影检查,现将结果报告如下.     

藤黄酸抑制前列腺癌PC-3细胞增殖并诱导其细胞凋亡

目的:研究中药藤黄的有效成分藤黄酸(gambogic acid,GA)对前列腺癌PC-3细胞增殖和凋亡的影响。方法:采用不同浓度的GA作用前列腺癌PC-3细胞后,在体外通过CCK-8比色法分析细胞的增殖情况,吖啶橙/溴化乙啶(acridine orange/ethidium bromide,AO/EB)双重染色法和FCM法分析细胞的凋亡情况;蛋白质印迹法检测细胞中凋亡相关蛋白P53、Bax和Bcl-2的表达变化。结果:GA不仅能抑制PC-3细胞的增殖,而且能有效诱导其细胞凋亡,与对照组比较差异有统计学意义(P<0.05),并且其抑制增殖和促凋亡作用呈浓度依赖性。CCK-8法检测结果表明,GA浓度>1μmol/L时,细胞的增殖能力受到明显抑制。AO/EB染色法显示,GA处理后的前列腺癌PC-3细胞核呈致密浓染橘红色,并伴有核浓缩和偏向。FCM法检测结果显示,GA处理后的前列腺癌PC-3细胞凋亡峰明显。蛋白质印迹法进一步表明,GA能够上调PC-3细胞中Bax和P53的表达水平,下调Bcl-2表达水平。结论:GA对前列腺癌PC-3细胞具有明显的抑制增殖和诱导凋亡作用。     

双侧上尿路结石并发肾功能不全术后急性肾衰的处理

目的：探讨治疗双侧上尿路结石并发肾功能不全术后急性肾衰的方法。方法：对54例双侧上尿路结石并发肾功能不全术后急性肾功能衰竭（ARF）患者的治疗方法进行回顾性分析,根据不同病情分别行急诊输尿管镜碎石（URL）和输尿管逆行插管或经皮肾造瘘（PCN）后二期碎石。所有病例监测血肌酐（Cr）和尿素氮（BUN）。结果：术后全部患者血Cr和BUN进行性下降,恢复到正常43例,氮质血症6例,尿毒症4例,死亡1例。结论：双侧上尿路结石并发肾功能不全应早期诊断,选择恰当的外科治疗时机和方法,一旦术后出现ARF,要立即明确原因,进行治疗。     

沙培林联合羟基喜树碱膀胱灌注与单独灌注预防膀胱癌术后复发疗效对比观察

目的：评价联合应用沙培林（OK-432）和羟基喜树碱（HCPT）膀胱腔内灌注及单独应用预防膀胱癌复发的疗效。方法：128例膀胱乳头状移行细胞癌患者行经尿道电切术和膀胱部分切除术后,随机分为3组：联合治疗组36例,OK-432组42例,HCPT、组50例。膀胱癌切除术后开始灌注,每周1次,共8周;以后每月1次,随访18个月。每2个月复查1次膀胱镜和尿细胞学检查。结果：联合应用组无复发,OK-432组3例复发,复发率7．14％（3／42）,HCPT组5例复发、复发率10．00％（5／50）。OK-432组与HCPT组比较差异无统计学意义（P〉0．05）;但二者与联合组比较,组间差异有统计学意义（P〈0．05）。三组均无严重不良反应和并发症。结论：联合应用OK-432、HCPT疗效优于单独应用,预防复发疗效好,不良反应不明显,有较高临床应用价值。     

The Relationship of Expression of bcl-2, p53, and Proliferating Cell Nuclear Antigen （PCNA） to Cell Proliferation and Apoptosis in Renal Cell Carcinoma

To investigate the relationship of bcl-2, p53, proliferating cell nuclear antigen (PCNA) to cell proliferation, apoptosis and pathological parameters, the patterns of cell growth and turnover in renal cell carcinoma (RCC), formalin-fixed and paraffin-embedded tissue blocks from 34 patients with RCC were examined. Cell proliferation activity was detected by PCNA immunostaining and the proliferation index (PI) was expressed as a percentage of the PCNA-positive cells in the tumor cells. Apoptosis was detected by terminal deoxy- nucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL), and the apoptotic index (AI) was expressed as a percentage of the TUNEL-positive cells in the tumor cells. Expressions of bcl-2 and pS3 were assessed immunohistochemically. Our results showed that the PI ranged from 6.0 % to 24.0 % (median 12.3 %) and the AI from 2.0 % to 8.0 % (median 5.4 ~) in RCC. The expression of the bcl-2 protein was demonstrated in 15 cases (44. 1%); the expression of the p53 protein, however, was seen in only 3 case. bcl-2 positivity was not associated with PI or AI or any pathological parameters. There were close associations between PI and tumor grade and stage, and a significant relationship between AI and the tumor grade of RCC. Our study suggests that bcl-2 positivity was not associated with PI or AI or any pathological parameters. There are close associations between PI and AI and tumor grade and stage of RCC. Active cell proliferation may be accompanied by frequent apoptosis in RCC.     

膀胱移行细胞癌复发风险多因素分析

目的：探讨与膀胱移行细胞癌复发时间相关的显著性因素，建立预测复发的数学模型。方法：应用Cox比例风险模型和Logistic线性回归方法对212例行手术治疗的膀胱移行细胞癌的临床表现、治疗、病理特点等方面16项指标与复发时间进行统计分析。结果：区域淋巴结转移是与复发时间相关的最显著因素，相对危险(Hr)为6．6，其次为多发性肿瘤(Hr=2．255)、三角区及膀胱颈肿瘤(Hr=2．053)、病理分期(Hr=2．057)、病理分级(Hr=1．569)；膀胱灌注治疗(Hr=0．559)和血尿程度(Hr=0.762)为保护性因素。据此建立影响膀胱癌术后复发的预测方程：PI=0．81305X4 0．71935X6(1) 0．36979X7 0．4507X8 0．72117X9 1．887llX10 (-0．58079)X14 (-0．2713)X15。代入原始数据，计算预测值，并将预测结果与实际结果比较，其灵敏度、特异度、及符合率分别为83．5％、67．6％、80．1％，两者基本相符。结论：根据上述模型可对移行细胞癌患者的预后做出较为准确的评价。