男性不育伴乙型肝炎病毒感染患者精子DNA完整性的初步研究

目的:探讨男性不育伴乙型肝炎病毒感染患者精子DNA完整性。方法:应用精子染色质结构分析检测90例男性不育伴乙型肝炎病毒感染者(A组)、82例男性不育者(B组)及70例正常生育者(C组)的精子DNA完整性:DNA碎片化指数(DFI)和高DNA可染性(HDS),分别比较3组之间精子DNA完整性的差异。结果:精子DNA完整性:DFI:A组为(28.17±13.06)%,高于B组(26.64±9.79)%和C组(15.67±4.73)%,A组及B组分别与C组比较差异有统计学意义(P<0.05),A组与B组比较差异无统计学意义(P>0.05);HDS:A组为(10.83±5.60)%,高于B组(9.04±3.48)%及C组(8.04±2.25)%,A组与C组比较差异有统计学意义(P<0.05)。结论:男性不育患者的精子DNA完整性与正常生育者存在差异,若伴乙型肝炎病毒感染,会进一步加重精子DNA损伤,主要表现为精子核成熟异常。     

灯盏细辛对大鼠肾冷缺血再灌注损伤中细胞凋亡的影响

目的 观察中药灯盏细辛对大鼠肾脏冷缺血再灌注损伤(IRI)中肾脏细胞凋亡及相关基因表达的影响.方法 封闭群SD大鼠36只,随机分为3组,每组12只.假手术组(A组),对照组(B组),实验组(C组).药物应用:C组术前15 min,灯盏细辛注射液按1.2 ml/100 g通过尾静脉注射,A、B组按相应剂量注射生理盐水.动物手术:A组,切除右肾.B、C组采用的是冷IRI模型,3组大鼠均在术后24h再次手术切除左肾进行检测.透射电镜检查肾组织形态学,免疫组织化学检测凋亡相关的基因bcl-2与bax的表达,原位末端标记法(TUNEL)检测细胞凋亡.结果 (1)超微结构检查(透射电镜):A组结构正常;B组细胞呈损伤形态:线粒体肿胀,微绒毛减少,胞质内空泡形成,部分细胞核可见凋亡迹象.C组病变较B组显著减轻.(2)免疫组织化学蛋白阳性染色指数(PI):缺血再灌注后B、C组的bcl-2表达分别为(21.21±1.18)％和(35.52±1.94)％,较A组(4.95±0.77)％均增多(P＜0.05),B组低于C组(P＜0.05).B、C组的bax表达分别为(58.55±2.90)％和(45.90±3.14)％,较A组(4.67±0.67)％增多(P＜0.05),而且B组高于C组(P＜0.05).A组的蛋白阳性染色指数的比值bcl-2/bax为(1.06±0.07)高于B组(0.35±0.03)和C组(0.78±0.07,P＜0.05),而且C组高于B组(P＜0.05).(3)细胞凋亡检测(TUNEL):细胞凋亡指数B组(28.57±3.58)％和C组(19.99±3.37)％均显著大于A组(2.33±0.42)％(P＜0.01),C 组小于B组(P＜0.01).结论 灯盏细辛减少大鼠肾脏冷IRI诱导的肾脏细胞的凋亡,与调节凋亡相关基因bcl-2与bax表达有关.     

地黄多糖干预对多次低剂量X线照射小鼠精子DNA完整性的影响

目的探讨地黄多糖干预对X线照射小鼠精予DNA完整性的影响。方法选择雄性昆明小鼠70只(5~6周龄;体重22±2g),随机分为A组(单纯照射组21只)、B组(照射+200mg/Kg RPS灌胃组21只)、C组(照射+800mg/Kg RPS灌胃组21只)、D组(对照组7只):再将A、B、C组分别随机分为3组,每组7只。所有小鼠在照射前5天开始每天灌胃1次,其中A组及D组给予0.2mL生理盐水灌胃,B组及C组分别给予200mg/Kg、800mg/Kg地黄多糖溶于0.2mL生理盐水灌胃,共灌胃24次。A、B、C组采用低剂量X线每天1次全身照射,共15次;单次照射剂量分别为0.025Gy、0.075Gy、0.100Gy,剂量率为0.025Gy/min。所有小鼠在末次照射结束后48h内脱颈法处死,立即收集附睾中精子并通过单细胞凝胶电泳(彗星试验)检测精子DNA的完整性。结果单纯X线照射后造成精子DNA损伤,其严重程度随剂量增加而加重。给予200mg/Kg及800mg/Kg地黄多糖灌胃则均能减轻低剂量X线照射对小鼠精子DNA的损伤,且后者较前者作用更明显,两者具有统计学意义(P0.05)。结论低剂量X线多次全身照射可引起小鼠精子DNA损伤,并呈剂量依赖性,不同剂量地黄多糖对X线照射后的小鼠精子DNA完整性具有保护作用。     

聚髌器联合克氏针内固定用于髌骨骨折治疗效果及安全性研究

目的 了解聚髌器联合克氏针用于髌骨骨折内固定的临床疗效及安全性.方法 纳入80例髌骨骨折为研究对象,根据内固定方法分为三组:A组行聚髌器联合克氏针内固定,30例;B组行克氏针钢丝张力带内固定,35例;C组行聚髌器内固定,15例.评价不同固定方法术后骨折愈合时间、膝关节功能及并发症.结果 A组、B组及C组患者骨折愈合时间分别为(11.6±2.0)周、(11.2±2.4)周与(13.1±2.1)周(P=0.023),A组、B组显著低于C组(P<0.05),A组与B组不存在统计学差异(P>0.05).A组、B组及C组患者术后6个月的膝关节功能优良率分别为86.7％、88.6％与66.7％(P=0.038),A组、B组显著高于C组(P<0.05).A组、B组及C组患者术后6个月膝关节屈曲角度平均值分别为(133.2±9.4)°、(135.6±7.5)°与(123.7±10.2)°(P<0.001),A组、B组显著高于C组(P<0.05),A、B组不存在统计学差异(P>0.05).A组患者术后疼痛、上下楼梯困难、下蹲困难的发生比例最低,显著低于B组与C组(P<0.05).结论 聚髌器联合克氏针行髌骨骨折复位内固定术可提供良好的内固定效果,同时降低术后并发症的发生风险.     

七氟醚不同吸入方法在小儿全身麻醉诱导中的效果观察

目的 对3种七氟醚吸入方法用于小儿全身麻醉诱导的麻醉效果进行对比.方法 选择扁桃体、腺样体摘除择期手术患儿90例,ASA分级Ⅰ级,按照随机数字表法分为逐步递增法组(A组)、潮气量法组(B组)、肺活量法组(C组),每组30例.吸入七氟醚160 s后行静脉穿刺,之后行进一步全身麻醉诱导.观察3组患儿入亲情诱导室(T0)、睫毛反射消失时(T1)、吸入160s后静脉穿刺时(T2)的MAP、HR、SpO2,意识消失时间、穿刺时体动反应发生率,麻醉诱导期间患儿合作程度、有无体动反应及副作用,手术结束后患儿自主呼吸恢复时间(Tc)、拔管时间(Td),并行躁动评分.结果 患儿意识消失时间A组为(58.1±0.3)s,B组为(41.2±0.3)s,C组为(33.8±0.8)s,A组最长而C组最短(P＜0.05);穿刺时体动反应发生率A组(96.7％)高于B组(16.7％)、C组(13.3％)(P＜0.05);诱导期体动反应发生率A组(100％)高于B组(13.3％)、C组(10％)(P＜0.05),仅在A组发现患儿发生心动过缓和唾液分泌各1例;诱导期患儿合作程度评分B组(0.20±0.07)分最低,A组(0.77±0.13)分次之,而C组(1.43±0.13)分最高,差异有统计学意义(P＜0.05).结论 在保证血流动力学稳定及不增加副作用发生的前提下,潮气量法吸入8％七氟醚更易为小儿所接受,更加适用于小儿全身麻醉诱导.     

基因组DNA和结缔组织生长因子基因启动子甲基化状态与2型糖尿病肾病的关系

目的 探讨糖尿病肾病(DN)患者基因组DNA及结缔组织生长因子(CTGF)启动子甲基化在DN发病中的作用.方法 根据1999年WHO糖尿病诊断标准,选取我院2型糖尿病患者90例及同期健康体检者30例,分为糖尿病组48例、DN组42例和健康对照组30例.提取各组受检者外周血白细胞DNA,应用高效液相色谱法检测基因组DNA整体甲基化水平;甲基化特异性PCR及测序方法检测CTGF启动子甲基化状态;用酶联免疫吸附法检测各组血清CTGF蛋白水平.结果 DN组基因组DNA甲基化水平为5.23％±0.09％,与糖尿病组(4.71％±0.03％)和健康对照组(4.37％±0.01％)比较,差异均无统计学意义(P＞0.05).DN组CTGF基因启动子甲基化水平为22.02％±12.90％,明显低于糖尿病组(49.18％±8.01％,P=0.019)及健康对照组(72.18％±19.30％,P=0.000),差异均有统计学意义.DN组血清CTGF蛋白水平为(193.44±11.9) mg/L,显著高于糖尿病组[(127.65±10.30) mg/L,P=0.031]及健康对照组[(95.84±5.1) mg/L,P=0.001].结论 DN患者CTGF启动子的甲基化程度明显低于糖尿病非肾病患者,而CTGF表达则高于非肾病患者,提示CTGF基因启动子的低甲基化可能参与DN的发生发展.     

补体在猪到猴异种心脏移植中的作用及机理

目的 探讨补体在异种大动物猪到猴心脏移植排斥反应中的作用及机理.方法 以梅山猪为供者,中国猕猴为受者,行异种腹腔异位心脏移植.随机将受者分为3组.A组(5只):为空白对照组,受者心脏移植后不作任何处理.B组(5只):为照射预处理组,受者于心脏移植前28 d、即1.5个月龄时接受60Coγ3 Gy全身剂量照射,其余同A组.C组(8只):为照射+胸腺注射预处理组,心脏移植前21 d,将供者的脾细胞(按照5×107个/只的数量)注入受者的两侧胸腺内,其余同B组.观察心脏移植术后各组移植心的存活时间;猪对猴单向混合淋巴细胞培养的刺激效应;采用双抗体夹心法检测补体C3和CD46的血清浓度;通过流式细胞术检测受者外周血细胞表面IgM、IgG阳性细胞百分比水平.结果 A、B、C三组移植心的存活时间分别为:(36.6±5.8)h、(65.6±6.5)h和(91.1±22.8)h,C组移植心的存活时间明显延长,与A组比较,P＜0.01,与B组比较,P＜0.05.C组在猪对猴单向混合淋巴细胞反应中的刺激效应较A、B组明显下降(P＜0.01).B、C组移植前补体水平(C3)无明显变化,但随着IgM、IgG水平的上升,发生排斥反应时C3和CD46水平显著降低.C组猕猴特异性抗猪抗体IgM及IgG的上升速度均较A、B组明显延缓.结论 对受者进行异种胸腺注射联合全身照射预处理在抑制T淋巴细胞免疫及体液免疫方面有重要作用,但无法抑制异种排斥反应中补体的激活,补体通过经典途径参与了延迟性异种排斥反应的发生.     

喉镜辅助光棒在困难气管插管中的临床应用

目的 探讨喉镜辅助下光棒在困难气管插管中的临床应用效果.方法 全麻插管手术患者120例,Yamamoto气道评级Ⅲ～Ⅳ级,随机均分为三组:喉镜组(A组)、单纯光棒组(B组)及喉镜辅助光棒组(C组).记录患者插管前后MAP、HR、去甲肾上腺素(NE)和肾上腺素(E)的变化,及插管成功率、插管时间、插管后并发症的发生情况.结果 与诱导后比较,插管即刻和插管后1min三组患者MAP、NE和E均显著升高,HR明显增快(P＜0.05),且A组高于B、C两组(P＜0.05),B、C两组相比差异无统计学意义.插管成功率A组为82.5％,B组为77.5％,C组为100％,C组显著高于A、B组(P＜0.01).插管时间A组为(43.3±12.5)s,B组为(47.1±15.1)s,C组为(31.1±10.7)s,C组显著短于A、B组(P＜0.05).结论 喉镜辅助下光棒在困难气管插管中成功率高、时间短,对血流动力学影响较轻,术后并发症少.     

红花黄素腹腔注射联合BMSCs移植对大鼠脊髓损伤的保护作用

目的 探讨红花黄素腹腔注射联合骨髓阳J充质干细胞(BMSCs)移植对大鼠脊髓损伤保护作用及其机制.方法 体外分离、培养BMSCs;SD大鼠45只,改良Allen法制备脊髓损伤模型,随机分为3组:A组、B组、C组.A组腹腔注射红花黄素治疗(40 mg/kg),B组和C组注射等体积的生理盐水,每日1次,共14 d;A组和B组损伤局部行BMSCs移植(6×106/60μl),C组注射等体积的PBS液.于术后1 d、2 d、3 d、4 d、7 d取材检测损伤处超氧化物歧化酶(SOD)、丙二醛(MDA)和细胞凋亡,术后1 d和7 d、14 d、21 d、28 d采用改良BBB评分法进行行为学检查评分.结果 术后随着时间的推移,各组BBB评分得分均增高,术后28 d时A组得分为(19.33±1.37),B组为(15.83±1.17),C组为(9.0±1.41),A组高于B组和C组,B组高于C组,差异有统计学意义(P＜0.05).SOD活性在相同时间点,均有A组高于B组和C组,B组高于C组,差异有统计学意义(P＜0.05);MDA含量在术后2 d、3 d、4d A组低于B组和C组(P＜0.05),且有A组低于B组(P＜0.05).术后3 d时各组细胞凋亡达高峰:A组(72.83±13.38),B组(96.33±6.35),C组(219.33±28.64);之后各组细胞凋亡数逐渐减少,至术后7d:A组(3.67±3.72),B组(16.33±5.54),C组(86.17±12.27);除术后第1天,在相同时间点,细胞凋亡数有A组少于B组和C组,B组少于C组,差异有统计学意义(P＜0.05).结论 红花黄素和BM-SCs移植均能抑制损伤脊髓局部自由基生成、脂质过氧化和细胞凋亡,减轻继发损伤,两者联合具有累加协同效应,有助于脊髓损伤大鼠肢体运动功能的恢复.     

胸腔镜胸腺切除术治疗重症肌无力500例

目的 总结胸腔镜胸腺切除术治疗重症肌无力(MG)的经验,对比胸腺瘤和非胸腺瘤MG患者的术后效果,分析胸腔镜联合纵隔镜和单纯胸腔镜治疗非胸腺瘤MG的远期效果.方法 2001年至2011年,采用胸腔镜胸腺扩大切除术治疗MG患者500例,根据是否合并胸腺瘤和术中是否联合纵隔镜分组:胸腔镜胸腺瘤组(A组)118例,胸腔镜非胸腺瘤组(B组)301例,胸腔镜联合纵隔镜非胸腺瘤组(C组)81例.结果 全组无术中死亡.完全电视胸腔镜下完成手术495例.A组手术(128.5±77.8) min,术后24.6％患者出现肌无力危象;B组手术(111.3±31.6) min,术后11.0％患者出现肌无力危象;C组手术(145.0 ±71.6) min,术后9.9％患者出现肌无力危象.术后随访3个月至11年,A、B、C组患者完全缓解率(CSR)分别为28.7％、37.3％、36.5％.无病生存曲线示术后3年B、C组CSR高于A组,术后5年C组CSR高于B组.术后第5年C组CSR接近60％,B组为50％,而A组仅为36％.结论 胸腔镜基础上联合纵隔镜胸腺扩大切除手术清除颈部、前纵隔脂肪组织及异位胸腺更为彻底,远期效果更加理想.与非胸腺瘤MG患者相比,胸腺瘤MG患者术后远期效果较差.     

Cystic fibrosis and the phenotypic expression of autosomal dominant polycystic kidney disease

Recent experiments in cultured cyst epithelial cells from kidneys of patients with autosomal dominant polycystic kidney disease (ADPKD) have shown that the cystic fibrosis (CF) transmembrane conductance regulator (CFTR) is present in the apical surface of these cells and mediates chloride (Cl-) and fluid secretion in vitro. To determine whether the presence of CF with the expression of mutated CFTR proteins modifies cyst formation in ADPKD, we studied a large family with both inherited diseases. ADPKD in this family is linked to PKD1. The family is composed of 26 members; 11 members with ADPKD, 4 members with CF, and 2 members with both diseases. Renal volumes measured by computerized tomography (CT), calculated creatinine clearances, and other clinical parameters in the family members with ADPKD and CF were compared with those in the family members with ADPKD alone, as well as to a large population of patients with ADPKD. The patients with CF and ADPKD, but not the CF heterozygote carriers with ADPKD, had less severe polycystic kidney and liver disease, as indicated by normal renal function; smaller renal volume, even when corrected for height and body surface area; and the absence of hypertension and liver cysts. These observations suggest that the coexistence of CF may reduce the severity of ADPKD.     

Evaluation of colonic diverticular disease in autosomal dominant polycystic kidney disease without end-stage renal disease.

A previous study had shown an increased prevalence (83%) of diverticula among patients with autosomal dominant polycystic kidney disease (ADPKD) with end-stage renal disease (ESRD) compared with other ESRD patients without ADPKD (32%). Others have also suggested an increased risk for diverticular complications in renal transplant recipients with ADPKD. To determine whether there was an increased occurrence of diverticula among non-ESRD patients with ADPKD, we studied 55 patients with ADPKD who were not receiving renal replacement therapy compared with 12 unaffected family members (non-ADPKD) and 59 random patients who had undergone barium enemas (control [C]). No study patient had a history of diverticular disease. All patients underwent a double-contrast barium enema after administration of glucagon. The occurrence, number, location, and size of diverticula were noted. There was no significant difference among the three groups in regard to sex (men: ADPKD, 42% versus non-ADPKD, 42% versus C, 37%) or age (ADPKD, 49.3 +/- 0.7 versus non-ADPKD, 51.2 +/- 2.1 versus C, 49 +/- 1 years). There was no significant difference in the percentage of patients with diverticula (ADPKD, 47% versus non-ADPKD, 58% versus C, 59%), the percentage with only right-colon diverticula (ADPKD, 5% versus non-ADPKD, 17% versus C, 5%), the mean number of diverticula in patients with diverticulosis (ADPKD, 13.8 versus non-ADPKD, 7.9 versus C, 9.9 diverticula), or the size of the largest diverticula (ADPKD, 9.5 versus non-ADPKD, 10.4 versus C, 10.5 mm). There was no significant difference in these variables between the patients with ADPKD with a creatinine clearance greater than 70 mL/min/1.73 m(2) (n = 25) or less than 70 mL/min/1.73 m(2). This study does not show the greater prevalence of diverticular disease in non-ESRD patients with ADPKD compared with the general population. Thus, patients with ADPKD need not be considered at greater risk for diverticular disease than the general population.     

Flow cytometric analysis of the DNA content in the urinary bladder cancers treated by radical cystectomy and pre-operative irradiation]

The DNA ploidy of bladder cancers treated by radical cystectomy following pre-operative irradiation was analyzed by flow cytometry using paraffin embedded samples. The DNA ploidy and its changes by irradiation were studied. We used flow cytometry in 30 patients with transitional cell carcinoma of the bladder who received pre-operative irradiation (40 Gy in 24 patients, 20 Gy in 5 patients and 60 Gy in one) with follow-up for at least 3 years. Total 140 paraffin embedded samples (4.6 samples per one patient) were available. The effects of therapy were related to the DNA patterns before irradiation and to the DNA ploidy changes after irradiation. 1. Eight DNA diploid tumors and twenty-two DNA aneuploid ones were detected before irradiation. Although diploid group didn't change its DNA ploidy after irradiation, of 22 aneuploid tumors 18 were changed to DNA diploid and 4 were not changed in their ploidy. 2. The tumor eradicating effect of irradiation was shown to be higher (p < 0.05) in the diploid group (5 of 8, 63%) than in the aneuploid group (5 of 22, 23%). 3. Overall survival rates were discussed in 3 groups (A, B and C), the group A was 10 of tumor free and 3 diploid tumors after irradiation, the group B was 13 of aneuploid tumors which changed to diploid ones and the group C was 4 of persistent aneuploid tumors. Each of 5 year survival rate was 100% (A), 58% (B) and 0% (C). Overall survival for C group was significantly shorter than for other groups (p < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)     

Deceased donor kidney transplantation in autosomal dominant polycystic kidney disease: a single-center experience

Renal transplantation (RTx) has become the treatment of choice for end-stage renal disease (ESRD) in autosomal dominant polycystic kidney disease (ADPKD), the most common genetic kidney disease. Because of the inheritable nature of this disease, live related donors might be avoided due to the fear of future appearance of polycystic disease. This retrospective singlecenter study was undertaken to evaluate patient/graft survival function vis-a-vis serum creatinine (SCr), rejection episodes and mortality in ADPKD vs matched control patients. Between 2000 and 2009, 18 (7.4%) deceased donor renal transplant (DDRTx) were performed for ESRD due to ADPKD. Diagnosis of ADPKD was established by family history and ultrasound. An individualized approach was applied for the need of pre-transplant nephrectomy. All recipients received rabbit-anti-thymocyte globulin induction and maintenance triple immunosuppression. Delayed graft function was observed in 33% patients, and 16% had biopsy-proven acute rejection. Over mean follow-up of 4.67 ± 2.2 years, patient and graft survival rates were 72.22% and 83.33%, with mean SCr (mg/dL) of 1.44 ± 0.54, 1.78 ± 0.42 and 2.2 ± 0.6 at 1, 5 and 10 years. Overall, 44.4% (n-8) underwent pre-transplant nephrectomy. Infection and cardio/cerebrovascular events were the main causes of death. Patient, graft survival and acute rejection were similar between ADPKD and control group. DDRTx in ADPKD has acceptable patient and graft survival. Because of the inheritable nature of the disease, and unavailability of genetic linkage analysis as a routine, DDRTx is a viable option to avoid using unrelated donors.     

常染色体显性遗传性多囊肾病患者与血管紧张素转换酶基因多态性的关系

目的 研究人常染色体显性遗传性多囊肾病（ADPKD）与血管紧张素转换酶（ACE）基因多态性的关系。 方法 用PCR方法对103例ADPKD患者及16个ADPKD家系（患者35例,非患病直系亲属30人）进行ACE基因多态性分析。收集患者及家系成员的临床资料,以发病年龄、肝囊肿、高血压、尿路感染、尿路结石、血尿等为主要参数,用统计学方法研究该病ACE基因多态性与ADPKD的关系。 结果 DD型患者的发病年龄比DI型患者早7.2岁[（31.90±11.41）岁比（39.10±10.08）岁];DD型患者的发病年龄比Ⅱ型患者[（46.15±14.74）岁]早14.25岁;DI型患者的发病年龄比Ⅱ型患者早7.05岁,各型间的差异均有统计学意义（均P ＜ 0.05）。3组间高血压、血尿差异有统计学意义。11个家系检查结果显示,ACE基因多态性在ADPKD家系中具有遗传连锁关系,但无统计学意义;家系中患病与非患病者ACE基因型频率差异无统计学意义;家系中患病与非患病者男女之间ACE基因型频率差异无统计学意义;家系中肾功能不全组与肾功能正常组之间DD型及D等位基因频率差异有统计学意义（P ＜ 0.05）。 结论 DD型患者的发病年龄较早,Ⅱ型患者的发病年龄较晚,DI型居中。DI型患者血尿的发生率较高,Ⅱ型患者血尿的发生率较低。DI型患者高血压的发生率较高。ACE基因多态性在ADPKD家系中不提供基因诊断信息; ACE基因多态性与人ADPKD的发病无显著相关性;ACE基因多态性与性别无明显关系。DD型基因型是ADPKD发生肾功能不全的易感因素。     

Linkage exclusion between the autosomal dominant polycystic kidney disease locus and chromosome 16 markers in a new family.

A family segregating for autosomal dominant polycystic kidney disease (ADPKD) is reported. The clinical picture was typical for ADPKD in some family members, although others showed mild involvement. DNA from family members was probed with seven chromosome 16 single-copy DNA sequences that mapped to the telomere of the short arm of the chromosome. The most likely order of six of the probes from the telomere is palpha3'HVR.64 at the designated locus D16S85, CRI-0327 at D16S63, CRI-090 at D16S45, CRI-0129 at D16S56, CRI-0133 at D16S58, and CRI-0136 at D16S60, with the PKD1 locus for ADPKD between D16S85 and D16S63. The seventh probe 24-1 at D16S80 had not been ordered in relation to the other sequences, but PKD1 had been mapped between it and D16S85. The three probes that were informative in our family, palpha3'HVR.64, CRI-090, and CRI-0136 had been linked to the disease locus at recombination frequencies of 4% and approximately 6 and 12%, respectively. Linkage was excluded between the ADPKD locus in our family and palpha3'HVR.64 at a recombination value of up to 6%. Linkage was also excluded between CRI-090 and the disease locus at a recombination value of up to 5%. The data for linkage between CRI-0136 and the ADPKD locus in our family were inconclusive. Multipoint analysis excluded the possibility that the disease in this family lies between the flanking genetic markers that have previously been used to define the genetic interval in which the most common form of polycystic kidney disease, PKD1, lies. We have not made a positive assignment of the ADPKD mutation in this family.(ABSTRACT TRUNCATED AT 250 WORDS)     

Influence of the endothelial nitric oxide synthase polymorphism on the progression of autosomal dominant polycystic kidney disease and IgA nephropathy

BACKGROUND: The reason of variability of clinical course and progression to end-stage renal failure (ESRF) of two widespread chronic nephropathies--autosomal dominant polycystic kidney disease (ADPKD) and IgA nephropathy (IGAN) is not clear. The endothelial dysfunction is considered in the number of factors possibly influencing the prognosis of these nephropathies. Our study tried to verify the hypothesis that endothelial nitric oxide synthase (ecNOS) gene polymorphisms in intron 4 could have some relevance to the progression of ADPKD and/or IgA nephropathy. METHODS: We examined 128 Czech patients with ADPKD (62 males, 66 females) and 93 patients with IGAN (51 males, 42 females). As a control group, we used 100 genetically unrelated healthy subjects (50 men, 50 women, mean age 51.2 +/- 8.2). The genomic DNA was amplified by polymerase chain reaction (PCR) and the products were separated on 1.5% agarose gel and visualized by ultraviolet transillumination. We compared homozygous subjects for ecNOSb allele with homozygous and heterozygous subjects for ecNOSa allele. RESULTS: The frequencies of ecNOSa/b + a/a and ecNOSb/b genotypes were 19% (19/100) and 81% (81/100) in the control group. The frequencies of ecNOSa/b + a/a and ecNOSb/b genotypes in ADPKD patients were: 26.6% (8/30) and 73.4% (22/30) in ADPKD patients with normal renal function, 30% (9/30) and 70% (21/30) in ADPKD with ESRF, 35.2% (18/51) and 64.8% (33/51) in young ADPKD patients, 60% (12/20) and 40% (8/20) in ADPKD patients with ESRF later than in 62 years. In IGAN, the frequencies of ecNOSa/b + a/a and ecNOSb/b genotypes were 24% (12/50) and 76% (38/50) in IgA with normal renal function and 20.9% (9/43) and 79.1% (38/43) in IgA with ESRF. CONCLUSION: Both in ADPKD and IGAN groups, there was no significant difference in the frequencies of ecNOS genotypes between patients with normal renal function and age matched patients with ESRF and between patients with normal renal function and control group. The frequency of ecNOSa allele was significantly higher in a number limited group ADPKD patients with ESRF later than in 62 years (Chi-square test p < 0.05). This higher frequency of a allele among ADPKD patients with later onset of ESRF could suggest the trend of positive influence of a allele in ADPKD patients.     

Mutations of the PKD2 gene in Taiwanese patients with autosomal dominant polycystic kidney disease

BACKGROUND: Mutation analysis in the context of clinical phenotypes helps clarify the pathogenesis of autosomal dominant polycystic kidney disease (ADPKD). Over 78 PKD2 gene mutations have been reported in the literature, but few have been described from an Asian population. This study attempted to characterize PKD2 mutations and their clinical implications among Taiwanese. METHODS: Twenty unrelated ADPKD patients with uncharacterized genotypes were screened for mutations in the PKD2 gene via single-strand conformation polymorphism (SSCP) of PCR products from genomic DNA, using previously reported PCR conditions and primers. RESULTS: This study identified two novel mutations (C681A and 2136-2137delG) and one mutation (C2407T) previously reported in a Cypriot family. Overall, we found PKD2 mutations in 15% (three out of 20) of the ADPKD patients screened. The mutations included two nonsense mutations (Y227X and R803X) and one frameshift mutation (712-715X) that could all lead to premature termination of translation. The locations of mutations in this study spanned the entire PKD2 gene on exons 2, 11, and 13 without clustering and did not influence the renal disease severity. CONCLUSIONS: The study identified two novel mutations and one recurrent mutation of the PKD2 gene in 20 Taiwanese patients. The characteristics of the mutations in this study resemble those reported among Western populations.     

Influence of the endothelial nitric oxide synthase polymorphism on the progression of autosomal dominant polycystic kidney disease and IgA nephropathy

BACKGROUND: The reason of variability of clinical course and progression to end-stage renal failure (ESRF) of two widespread chronic nephropathies-autosomal dominant polycystic kidney disease (ADPKD) and IgA nephropathy (IGAN) is not clear. The endothelial dysfunction is considered in the number of factors possibly influencing the prognosis of these nephropathies. Our study tried to verify the hypothesis that endothelial nitric oxide synthase (ecNOS) gene polymorphisms in intron 4 could have some relevance to the progression of ADPKD and/or IgA nephropathy. METHODS: We examined 128 Czech patients with ADPKD (62 males, 66 females) and 93 patients with IGAN (51 males, 42 females). As a control group we used 100 genetically unrelated healthy subjects (50 men, 50 women, mean age 51.2 +/- 8.2). The genomic DNA was amplified by polymerase chain reaction (PCR) and the products were separated on 1.5% agarose gel and visualized by ultraviolet transillumination. We compared homozygous subjects for ecNOSb allele with homozygous and heterozygous subjects for ecNOSa allele. RESULTS: The frequencies of ecNOSa/b + a/a and ecNOSb/b genotypes were 19% (19/100) and 81% (81/100) in the control group. The frequencies of ecNOSa/b + a/a and ecNOSb/b genotypes in ADPKD patients were: 26.6% (8/30) and 73.4% (22/30) in ADPKD patients with normal renal function, 30% (9/30) and 70% (21/30) in ADPKD with ESRF, 35.2% (18/51) and 64.8% (33/51) in young ADPKD patients, 60% (12/20) and 40% (8/20) in ADPKD patients with ESRF later than in 62 years. In IGAN the frequencies of ecNOSa/b + a/a and ecNOSb/b genotypes were 24% (12/50) and 76% (38/50) in IgA with normal renal function and 20.9 % (9/43) and 79.1% (38/43) in IgA with ESRF. Conclusion: Both in ADPKD and IGAN groups there was no significant difference in the frequencies of ecNOS genotypes between patients with normal renal function and age matched patients with ESRF and between patients with normal renal function and control group. The frequency of ecNOS a allele was significantly higher in a number limited group ADPKD patients with ESRF later than in 62 years (Chi-square test p < 0.05). This higher frequency of a allele among ADPKD patients with later onset of ESRF could suggest the trend of positive influence of a allele in ADPKD patients.     

Effect of dilazep dihydrochloride on urinary albumin excretion in patients with autosomal dominant polycystic kidney disease

Proteinuria and microalbuminuria occur with a highly variable severity and are associated with progression of autosomal dominant polycystic kidney disease (ADPKD). Dilazep dihydrochloride, an antiplatelet drug, is effective in patients with immunoglobulin A nephropathy or diabetic nephropathy. We studied whether dilazep dihydrochloride affects the urinary albumin excretion (UAE) in normotensive and hypertensive patients with ADPKD. Twelve normotensive ADPKD patients with microalbuminuria were randomly assigned to two groups: a dilazep (300 mg/day) treatment group (n = 6, group A) and a placebo group (n = 6, group B). In addition, 10 hypertensive ADPKD patients with microalbuminuria were randomly assigned to two groups: a dilazep (300 mg/day) treatment group (n = 5, group C) and a placebo group (n = 5, group D). Treatment with dilazep was continued for a period of 6 months, at the end of which the UAE was reduced form 130 +/- 52 to 46 +/- 26 microg/min (p < 0.01) in group A. There was no reduction in group C. There were no changes in UAE in placebo groups B and D. These results suggest that dilazep dihydrochloride may be effective in reducing UAE in normotensive ADPKD patients with microalbuminuria.