Nitric oxide synthase activity in the human urogenital tract

Nitric oxide (NO) has been suggested as a non-adrenergic non-cholinergic neurotransmitter in the urogenital tract and has previously been shown to have a smooth muscle relaxing effect in the urogenital organs both in various animals and in humans. It has been shown that NO is a mediator of the erection and the dilatation of the bladder neck and urethra. The aim of the study was to analyse nitric oxide synthase (NOS) activity in the human urogenital tract. NOS activity was measured by the conversion of l-[U-¹⁴C] arginine to l-[U-¹⁴C] citrulline. In the upper urinary tract there was Ca²⁺-dependent NOS activity in the renal pelvis, but no significant NOS activity could be found in the ureter. In the lower urinary tract we found high Ca²⁺-dependent NOS activity in the urethra, intermediate activity in the bladder neck and comparatively low activity in the detrusor muscle. In the male genital tract the testis and epididymis had no significant NOS activity. The vas deferens, prostate, seminal vesicle and corpus cavernosum were found to have high levels of Ca²⁺-dependent NOS activity. Ca²⁺-independent NOS activity was not obtained in the urogenital tract. Our results correspond well with previous functional studies indicating NO to be an important nerve-induced mediator of erection and in the micturition reflex, but also suggest that NO may be involved in several other functions in the human urogenital tract.     

A possible role for nitric oxide in the regulation of human ureteral smooth muscle tone in vitro

There is ample evidence that nitric oxide (NO) is an important neurotransmitter in many tissues of the urogenital tract. The aim of the present study was to examine the possible role of NO in ureteral relaxation. Human ureteral rings were mounted in organ bath chambers and precontracted with KCl. Increasing doses of the NO donor linsidomine (SIN-1) were added with and without prior blockade of the NO/cGMP pathway by methylene blue and protein kinase (PK) inhibitors Rp-8-pCPT-cGMPS and Rp-8-CPT-cAMPS. Electrical field stimulation (EFS) was done before and after incubation with L-NOARD (N ^G-nitro-L-arginine) and TTX (tetratodoxin). For detection of neuronal NO synthase (NOS), ureters were stained immunohistochemically. Ureteral strips were dose dependently relaxed by SIN-1; preincubation with methylene blue and protein kinase G inhibitor significantly reduced the SIN-1-induced relaxations. No effects of L-NOARG and TTX on EFS-induced tone alterations were found. NOS-positive neuronal axons and nerve-ending-like structures were found in the muscular layers. Our in vitro findings suggest that ureteral relaxation may involve the NO pathway.     

体外培养的人腹主动脉瘤血管平滑肌细胞中一氧化氮的生成和诱生型一氧化氮合酶的表达

目的探讨体外培养的人腹主动脉瘤 (AAA)血管平滑肌细胞 (SMC)及其表达诱生型一氧化氮合酶 (iNOS)和培养液中生成一氧化氮 (NO)的情况。方法 0 0 2 %Ⅰ型胶原酶消化法进行AAA SMC原代培养 ,平滑肌α 肌动蛋白 (α SMA)鉴定SMC并绘制AAA SMC的增殖曲线 ;免疫细胞化学方法检测AAA SMC中iNOS蛋白的表达 ,并测定原代及 2代细胞培养液中亚硝酸盐和硝酸盐的浓度之和 (NO2 - NO3 -,NOX)。结果酶消化法成功培养AAA SMC ,α SMA阳性率为 4 5 %± 5 8% ,传代培养发现AAA SMC增殖力有限 ;AAA SMC中iNOS蛋白阳性率 86 7%± 4 6 % ,细胞培养液中存在高浓度的NOX。结论AAA SMC存在异常增殖但增殖力有限 ,且细胞可能存在表型变化 ;AAA SMC中iNOS蛋白的高表达及NO的过量生成 ,提示由SMC生成的过量NO可能在腹主动脉瘤发病机制中具有重要作用。     

输尿管肾镜术处理上尿路血尿

目的　提高上尿路血尿的诊治水平。　方法　血尿病例 12 1例 ,经B超、KUB加IVU、CT及膀胱镜检查怀疑为上尿路来源 ,行输尿管肾镜进一步明确诊断并采取相应治疗措施。　结果　输尿管肾镜术对上尿路血尿的诊断率为 92 % (111/ 12 1) ,诊断输尿管中下段小结石 4 5例 ;肾盂输尿管肿瘤 32例 ,其中 <10mm者 3例 ;肾出血性疾病 19例 ;输尿管息肉 15例。 19例肾出血性疾病和 10例未发现明确病灶者治疗后随访 6个月～ 8年 ,长期成功率 79% (2 3/ 2 9)。　结论　应用输尿管肾镜术诊断和处理上尿路血尿效果良好 ,值得推广。     

双侧上尿路结石手术处理原则的探讨(附53例报告)

目的：探讨双侧上尿路结石的处理原则。方法：回顾性分析53例双侧上尿路结石患者的手术资料。结果：48例患者术后获得痊愈，其中17例肾功能不全患者，术后有12例血肌酐恢复正常，5例稳定于代偿期(血肌酐值138—313μmol／L)。结论：一期手术取石解除双侧梗阻有利于双侧分肾功能的恢复，应优先考虑。分期手术应根据肾脏对抗平衡理论，按总肾功能及分肾功能的不同分别对待。     

Optimizing management of upper tract urothelial carcinoma

Upper tract urothelial cancer (UTUC) is a rare cancer of the urothelium, comprising only a fraction of cases as compared to urothelial tumors of the bladder. As a result, systemic treatment approaches in bladder cancer are often applied to patients with UTUC. Given the anatomical location of these tumors, the age, the comorbid conditions of these patients with UTUC, and the need for radical nephroureterectomy for treatment, most patients have substantial impairment of renal reserve. There is growing evidence for the benefit of perioperative chemotherapy in this disease. Patients with UTUC have high rates of microsatellite instability and fibroblast growth factor receptor 3 mutations as compared to their bladder counterparts presenting unique, important subsets in UTUC. Immune checkpoint inhibitors targeting the programmed death receptor 1 and ligand have provided a new second-line treatment option for patients with UTUC and appear particularly well suited for patients with microsatellite instability. More work in understanding the molecular gene signatures and its relationship to response to chemotherapy, immunotherapy, and targeted therapy is needed to continually optimize care for patients with all stages of disease. Advances in UTUC are possible, when one accounts for the unique clinical and biological features of this disease.     

经皮肾穿刺造瘘在上尿路梗阻性疾病的应用

目的:探讨经皮肾穿刺造瘘在上尿路梗阻性疾病(UUTO)的应用价值.方法:对42例UUTO患者,先行经皮肾穿刺造瘘引流,待肾功能改善、机体状况好转或经引流及造影确定诊断之后,12例行经皮肾取石碎石术,11例行开放肾、输尿管切开取石术,4例行开放肾切除术,4例行肾盂输尿管成型(3例加行取石)术,3例经造瘘管注射硬化剂治疗,3例行输尿管肾盏吻合术,3例行输尿管狭窄段切除端端吻合术,1例行输尿管松解术,1例行输尿管皮肤造瘘术.结果:38例患者一次穿刺成功.全部病例均及时解除了上尿路梗阻,肾功能得到改善,或为进一步诊治创造了条件.在保肾治疗的35例,31例治愈出院;4例肾功能改善,维持在轻度氮质血症水平;4例肾切除患者和3例注射硬化剂治疗患者均治愈出院.结论:经皮肾穿刺造瘘安全、有效,在UUTO的诊治中具有重要的应用价值.     

上尿路腔内导管超声的临床应用评价

目的 评价腔内导管超声诊断上尿路疾病的可行性和有效性. 方法上尿路疾病患者68例.男30例,女38例.年龄17～81岁,平均58岁.采用频率10 MHz、外径8 F微探头腔内导管超声探测显示肾盂及周围病变48例,输尿管及周围病变20例;其中双侧检测2例,随访检测3例,共73例次.根据患者病变病理或临床随访结果评价腔内导管超声诊断的有效性.结果 68例患者中,成功检测62例(91%),其中肾盂及周围病变46例(96%)、输尿管及周围病变16例(80%).导管超声检出肾盂及周围病变异常41例(89%),其中经手术病理诊断28例、经临床随访和根据其他检查确诊7例、未被证实6例;检出输尿管及周围病变异常者12例(75%).20例输尿管及周围病变者中经手术病理诊断或临床随访和根据其他检查确诊18例,14例行输尿管手术,输尿管微结石4例采用保守治疗.结论 上尿路腔内微探头导管超声探测能清晰显示腔内病灶的形态、大小、回声和部位,而且能精细、准确、实时地显示腔外结构,为上尿路疾病诊断提供了一种新方法.     

金属支架植入治疗上尿路闭锁的中期结果

目的评价金属支架植入治疗上尿路闭锁的中期疗效。方法1995年10月至1998年12月，采用金属支架永久植入治疗上尿路闭锁患者13例，其中肾盂输尿管连接部闭锁8例，肾下盏输尿管吻合口闭锁3例，输尿管上段闭锁和输尿管膀膀吻合口闭锁各1例。闭锁长度1．0～3．6cm。采用影像学方法定期随访，必要时行输尿管镜检查。结果13例术后随访1～9年，平均92个月。输尿管引流通畅6例，需辅助停留输尿管内支架管、换管时见支架处上皮覆盖完全3例，因肾功能进行性恶化而行肾切除2例，因肾积脓感染无法控制而取出金属支架2例。3例患者分别于置管后4、6、6个月发现支架内肉芽组织生长，用钬激光汽化肉芽组织。其中1例4个月后又出现肉芽组织生长，再用钬激光汽化后长期留置双J管。2例患者于置管术后28、32个月并发支架近端结石，分别用微创经皮取石术和输尿管镜取出。结论金属支架植入治疗上尿路闭锁安全、有效，中期结果满意，其对上尿路动力学的影响尚需进一步研究。     

人内皮型一氧化氮合成酶基因重组腺病毒载体构建及其在人血管平滑肌细胞的表达

目的 利用AdMax腺病毒载体系统构建人内皮型一氧化氮合成酶基因(heNOS)重组腺病毒并检测其在体外培养人血管平滑肌细胞中表达.方法 将质粒PMSCV-heNOS通过聚合酶链反应(PCR)法扩增出heNOS全长基因,插入PSUCMV中构建成腺病毒穿梭质粒PSUCMV-heNOS,转化DH5α大肠杆菌,挑选细菌单克隆进行DNA测序,酶切鉴定正确;通过PSUCMV-heNOS与质粒pBHGE3在293细胞中同源重组,得到携带heNOS基因的复制缺陷型重组腺病毒载体(AdCMV-he-NOS).转染体外培养的人血管平滑肌细胞,MTT检测细胞增殖,用免疫组织化学和Western blot等方法检测heNOS蛋白的表达.结果 (1)PCR产物电泳结果,酶切鉴定和测序鉴定结果证实构建人eNOS重组腺病毒载体成功,病毒滴度达3.5×l010PFU/ml;(2)根据体外培养的人血管平滑肌细胞(HVSMCs)形态学变化,免疫组织化学进行平滑肌细胞表型标志物α-actin染色,证实为HVSMCs;(3)转染120 h,AdCMV-heNOS病毒感染复数(MOI)分别为50、150、250、300、450、A570值分别为1.410±0.081、1.357±0.150、1.303±0.311、0.995±0.248、0.731±0.101,其中感染复数300明显稳定抑制血管平滑肌细胞的增殖,差异有统计学意义(P＜0.01).免疫组织化学和Western blot检测显示转染细胞heNOS蛋白明显表达.结论 heNOS重组腺病毒的构建及其表达为血管内膜增生等疾病的基因治疗提供可能.

Abstract：

Objective To construct recombinant adenovirus vector carrying the human endothelial nitric oxide synthase gene (heNOS) and detect its expression in human vascular smooth muscle cells in vitro. Methods heNOS was cloned by polymerase chain reaction (PCR) using plasmid PMSCV-heNOS,heNOS cDNA was inserted into adenovirus shuttle plasmid-PSUCMV to generate a recombinant plasmid PSUCMV-heNOS, transfected into E. coli DH5α, and positive clones were correctly constructed and confirmed by DNA sequencing analysis and restriction endonucleas analysis. The plasmid PSUCMV-heNOS was co-transfected with pBHGE3 in 293 cells to generate replication-defective recombinant adenovirus vector carrying heNOS gene. Human vascular smooth muscle cells were transfected by Ad-heNOS. The effect on the proliferation of human vascular smooth muscle cells was investigated by MTT assay. The expression of heNOS was detected by immunohistochemistry staining and Western blotting. Results ( 1 ) PCR, restrictive digestion, and sequencing revealed the successful construction of the recombinant adenovirus vector carrying heNOS gene and its titer was 3.5 × 1010 PFU/ml; (2) It was confirmed that the human vascular smooth muscle cells cultivated in vitro were characterized by morphology and immunohistochemistry strain of α-actin; (3) At 120 h, the A570 values in multiplicity of infection (MOI) 50, 150,250,300,450 were respectively 1. 410 ±0. 081, 1. 357 ±0. 150, 1.303 ±0. 311,0. 995 ±0. 248 and 0. 731 ±0. 101. The proliferation of human vascular smooth muscle cells was significantly and stably inhibited with MOI 300 of AdCMVheNOS ( P ＜ 0. 01 ). Immunostaining and Western blotting with heNOS in engineered cells were positive.Conclusion The successful construction of AdCMV-heNOS and expression may provide an opportunity for the gene therapy of vascular intimal hyperplasia.     

孤立肾上尿路结石的ESWL治疗

目的:总结孤立肾并发上尿路结石ESWL治疗的经验。方法:电压比非孤立肾者略低,JT-Ⅲ型机8～12kV,HB-V型机4～8kV;放电次数比非孤立肾者略少,Ⅲ型机1500～2000次,V型机3000～3500次;同时减慢冲击频率;间隔时间二周以上比非孤立肾者稍长。直径大于2cm或多发结石且颗粒较大者,先行经皮肾镜取石后残石再行ESWL,多发或直径大于1.5cm的结石留置双J管后再ESWL,梗阻引起急性肾功能减退者急诊ESWL或先行肾造瘘或逆行插管引流积水,肾功能基本恢复后再ESWL。结果:22例独肾结石除7例多发外,15例一次成功,6例输尿管结石除1例再碎石外,5例一次成功。结论:ESWL治疗孤立肾上尿路结石损伤小且疗效好。     

Impact of surgical waiting time on survival in patients with upper tract urothelial carcinoma: A national cancer database study

Purpose

To evaluate the impact of surgical waiting time (SWT) on the survival outcome in patients with upper tract urothelial carcinoma (UTUC).

经皮肾镜技术治疗上尿路肿瘤的初步经验

目的 评价经皮肾镜技术在上尿路肿瘤中的应用效果.方法 2006年6月至2010年6月经皮肾镜治疗上尿路肿瘤患者8例(10侧).男6例(7侧),女2例(3侧).年龄52～72岁,平均61岁.孤立肾4例,慢性肾功能不全2例,双侧肾盂肿瘤2例.高级别肿瘤4侧,低级别肿瘤6侧.肿瘤直径0.5～3.5 cm,平均2.6 cm.患者均接受经皮肾镜激光或电刀肿瘤切除术,术中留置输尿管支架管,术后经肾造瘘管灌注化疗药物.结果 8例手术均获成功,手术时间45～95 min,平均73 min;术中出血量20～300 ml,平均50 ml,术后SCr水平较术前下降或无明显变化.随访10～36个月,采用CT、MRI及输尿管镜检观察肿瘤复发情况.1例死于肿瘤转移,2例肿瘤局部复发.余5例未见肿瘤复发.结论 经皮肾镜技术治疗上尿路肿瘤安全可行,手术效果良好,对不宜行肾输尿管切除术的上尿路肿瘤患者来说是一种良好的选择.

Abstract：

Objective To evaluate the application of percutaneous nephroscopy in the treatment of upper urinary tract transitional cell carcinoma, particularly renal pelvic carcinoma. Methods From June 2006 to June 2010, eight cases (with 10 sides) of renal pelvic carcinoma received percutaneous nephroscopy tumor resection. There were six males (with 7 sides) and two females (with 3 sides) in the study group. There were six cases with solitary kidney and two cases with bilateral renal pelvic tumors. There were four cases with high-grade tumors and six cases with low-grade tumors. The age of patients ranged from 52 to 72 yrs (average 61.2 yrs). Tumor sizes ranged from 0.5 to 3.5 cm (average 2.6 cm). Patients were treated with laser or electrocautery through percutaneous nephroscopy. A ureteral stent was placed in the patients after the procedure. Chemotherapy was administered postoperatively through the nephrostomy tube. Results All the operations were successfully completed uneventfully. The operative time was 45-95 min (average 73 min), estimated blood loss was 20-300 ml (average 50 ml). No remarkable differences were found in serum creatinine levels before and after operation. After 10 to 36 mon. follow-up by CT, MRI, and ureteroscopy, one patient died of tumor metastasis and two patients had local tumor recurrence. The remaining patients had no local recurrence. Conclusions Percutaneous nephroscopy in treating renal pelvic tumor is safe and feasible. This is a better choice for the renal pelvic carcinoma patients who are unsuitable for ureteronephrectomy.     

Osteopontin overexpression predicts poor prognosis of upper urinary tract urothelial carcinoma

Objectives

Studies indicate overexpression of osteopontin (OPN) promotes carcinogenesis, progression and metastasis of multiple human malignancies. However, the function of OPN in urothelial carcinoma (UC) of the upper urinary tract has not been investigated. This study evaluates the clinical significance of OPN expression in upper urinary tract UC.

Materials and methods

One hundred and ten cases (median age = 64, range = 24–84 years) of renal pelvic or ureter UC were retrospectively reviewed in this study. OPN expression were evaluated by immunohistochemistry staining on paraffin-embedded section of the tumor and scored by two qualified pathologists.

Results

High OPN expression was found in 54 (49.1%) of the cancer specimens. OPN expression was not significantly correlated with tumor T stage (P = 0.761), N stage (P = 0.339) or grade (P = 0.349). However, OPN expression was differently expressed by gender (P = 0.012) and cancer location (P = 0.026). OPN expression did not correlate with bladder recurrence-free (P = 0.661) or extra-bladder recurrence-free (P = 0.787) survival, but high OPN expression was a significant predictor for cancer-specific survival (P = 0.014).

Conclusion

Our findings indicated that higher OPN expression is a potential biomarker to predict patient survival. Further study is necessary to investigate the role of OPN in the carcinogenesis of upper urinary tract UC.     

急性上尿路梗阻性无尿31例报告

报告急性上尿路梗阻性无尿３１例，均行手术治疗，结果３１例出现多尿期，日尿量达４０００－７０００ｍｌ，后逐渐降至２５００－３０００ｍｌ，持续数月。并就本病的有关问题进行讨论，认为解除梗阻后肾功能恢复慢与否，首先取决于梗阻的时间长短，其次是肾脏的原来状况和梗阻后是否滥是利尿剂等；ＫＵＢ及Ｂ超检查是首选的诊断方法，治疗原则是及时解除梗阻，有效防治感染，最大限度地保护肾脏功能。     

Progress in urodynamic research on the upper urinary tract: implications for practical urology

The development of new surgical techniques for bladder substitution and continent urinary diversion has extended interest in urodynamics of the upper urinary tract. From a subdiscipline attracting mainly scientists and bioengineers, renal pelvic kinetics and ureteral peristalsis have evolved as important factors in routine clinical urology. The observed changes in peristaltic pattern during high diuresis, obstruction and urinary reflux have influenced management of stone disease and neurogenic bladder. The demonstration that high intravesical pressure is reflected to the kidney not only when the ureteric orifice is incompetent, but also during high diuresis, established the necessity for low pressures in neobladders. Much further clarification of urinary transport from the renal tubules to the bladder should be achievable by refined techniques of fluoroscopy, isotopic renography and manometry.     

脑出血患者一氧化氮、一氧化氮合酶的变化及临床意义

目的 探讨一氧化氮(NO)、一氧化氮合酶(NOS)与脑出血的关系。方法 测定76例脑出血患者及66例健康人血浆NO、NOS含量，分析上述指标在出血后3个时期的变化以及与患者颅内血肿量、是否合并蛛网膜下腔出血和临床预后的关系。结果 与对照组相比，脑出血1～3d组NO、NOS含量明显高于4～7d组、14d组和对照组，NO、NOS变化与颅内血肿量和／或合并蛛网膜下腔出血有一定关系。结论 NO、NOS参与了脑出血病理生理过程，其变化对病情的预后有一定的意义。     

输尿管镜下处理上尿路狭窄56例报告

目的探讨输尿管镜下处理上尿路狭窄的疗效. 方法采用输尿管镜下直视扩张、电刀切割、气囊扩张等方法处理上尿路狭窄56例. 结果 49例扩张成功,占87.5%(49/56);7例失败.无输尿管穿孔、断裂、大出血等并发症发生.43例随访0.5 ～ 3年,平均2.5年,狭窄消除33例,狭窄改善不明显6例,狭窄复发4例;肾积水消失30例,减轻5例,无改善8例. 结论输尿管镜下处理上尿路狭窄创伤小,疗效满意.     

双侧上尿路全程扩张的临床分析和处理

目的:探讨双侧上尿路全程扩张的病因、临床诊断、鉴别诊断和治疗方法。方法:回顾性分析近3年临床所见双侧上尿路全程扩张患者25例的临床资料,并结合文献复习予以讨论。结果:根据腔内、腔外梗阻及非梗阻因素所致双侧上尿路全程扩张的诊断,采取药物、自我导尿、留置输尿管支架管、微创或手术治疗等解除梗阻或非梗阻因素,均取得良好效果。结论:双侧上尿路全程扩张病因复杂,明确诊断十分重要,有利于选择合适的治疗方法。