Toxicity of ifosfamide,cyclophosphamide and their metabolites in renal tubular cells in culture

Ifosfamide (IF) and cyclophosphamide (CP) are highly effective alkylating cytostatic drugs. IF and CP have to be activated through a metabolic step in vivo; numerous metabolites are known. While both IF and its structural isomer CP have severe urotoxic side effects, only IF is also a nephrotoxic drug, causing tubular damage resulting in Fanconi syndrome in some cases. Little information is available regarding the pathogenic mechanism of tubular damage by IF. We used the renal epithelial cell line LLC-PK₁, which has many properties of the proximal tubule, in order to investigate the toxicity of IF and CP and of their reactive metabolites 4-hydroxy-IF (4-OH-IF), 4-hydroxy-CP (4-OH-CP), acrolein and chloracetaldehyde (CAA). Protein content of monolayers, DNA and RNA synthesis were determined by standard techniques (thymidine and uridine incorporation). IF and CP had the lowest toxicities of all compounds tested. Both drugs inhibited thymidine incorporation by about 30% at a concentration of 300 mol/l after 1 h incubation. 4-OH-IF and 4-OH-CP were significantly more toxic than the parent drugs. Thymidine incorporation, the most sensitive parameter, was reduced by about 70% by 300 mol/l of either compound. In addition, 4-OH-CP reduced the total protein content of monolayers. 4-OH-IF did not effect protein content and RNA synthesis. Acrolein, the most toxic metabolite tested, reduced all three parameters significantly at concentrations of 50–75 mol/l after 1 h. Incubation of cells with 100 mol/l of acrolein showed an effect after only 1 min. CAA significantly damaged monolayers with a reduction of total protein at a concentration of 50–100 mol/l. Thymidine incorporation was decreased only moderately by CAA, while uridine incorporation was stimulated, which may be interpreted to reflect a mechanism of repair. We conclude that both CP and IF and their metabolites are toxic in renal tubular cells in culture. CAA may play a larger role in the development of renal tubular damage after therapy with IF than was previously recognized.     

尿微量蛋白检测对移植肾早期功能损害的诊断价值

目的探讨尿微量蛋白检测对肾移植患者术后早期肾功能损害的诊断价值。方法 200例肾移植患者(肾移植组)按移植术后时间长短分层随机抽样,分为5组:术后1～3个月组,术后4～11个月组,术后1～5年组,术后6～10年组,术后10年以上组,每组40例。另随机抽取拟行肾移植术而进行术前检查的尿毒症患者20例为尿毒症对照组。用速率散射比浊法分别对6组患者的清洁新鲜中段尿进行尿微量蛋白测定与尿蛋白定性检测,同时采用全自动生化仪检测血清肌酐(Scr)。结果与术后1～3个月组比较,术后4～11个月组的尿α1-微球蛋白即开始升高(P<0.05),术后1～5年组、6～10年组尿微量白蛋白、尿转铁蛋白明显升高(均为P<0.05)。术后10年以上组尿微量白蛋白、尿α1-微球蛋白、尿转铁蛋白、尿免疫球蛋白(Ig)G均明显升高(均为P<0.05)。术后10年以上组尿蛋白阳性率明显高于其它时间组(均为P<0.05)。肾移植术后不同时间Scr水平比较差异无统计学意义(均为P>0.05)。结论尿微量蛋白的变化较尿蛋白定性及Scr的改变更为敏感,宜作为早期移植肾功能受损的常规监测指标。     

Assessment of silver-coated urinary catheter toxicity by cell culture

Summary The toxicity of silver-coated urinary catheters was assessed using a cell culture technique. The inhibitory effect of catheter extracts on the uptake of ³H-labelled thymidine by mouse fibroblasts was measured. The results show that silver-coating had no toxic effect whereas silvernitrate and silversulphate coating did have a toxic effect.     

肾移植术后泌尿系感染的特点分析

目的探讨肾移植术后泌尿系感染的特点。方法回顾性分析2009年1月至12月在暨南大学附属第一医院进行肾移植的38例患者(男性24例,女性14例)的临床资料,对其术后1年内尿常规、尿培养结果及其他临床资料进行总结分析。结果 38例患者肾移植术后1年内共查尿常规次数787例次,其中尿常规阴性结果为633例次,标本污染53例次,细菌尿次数101例次,其中71例次发生肾移植术后半年内。101例次泌尿系感染中,无症状性菌尿为68例次,上尿路感染19例次,下尿路感染14例次,并发脓毒血症者3例次。38例患者中,共有25例(男14例、女11例)患者出现细菌尿,有19例患者出现不少于2次泌尿系感染。最常见的致病菌为大肠埃希菌(36例次),其次是粪肠球菌(26例次)。肾移植术前血液透析患者术后感染率为70%。36例术后7d内拔出尿管和双J管的患者肾移植术后感染率64%,而2例肾移植术后7d后拔出尿管和双J管患者均发生感染。术后出现膀胱输尿管反流的3例患者均出现泌尿系感染。结论肾移植术后泌尿系感染具有高发性、反复发作的特点,最常见的致病菌为大肠埃希菌。女性、尿管和双J管的长时间留置、术后膀胱输尿管反流、术前长时间血液透析等都会诱发肾移植术后的泌尿系感染。     

氯沙坦对特发性膜性肾病患者尿蛋白谱的影响

目的观察氯沙坦对特发性膜性肾病(idiopathic membranous nephropathy,IMN)患者尿蛋白谱的影响,以期阐明其部分治疗机制。方法将79例IMN患者随机分为观察组(42例)及对照组(37例),所有患者在纳入实验前均常规治疗1个月。观察组在常规治疗基础上服用氯沙坦(100mg/d)治疗3个月,对照组继续给予常规治疗3个月。治疗前后分别进行尿β2-微球蛋白肌酐比值(urinaryβ2-microglobulin to creatinine ratio,β2-MG/Cr)、视黄醇结合蛋白肌酐比值(retinol binding protein to creatinine ratio,RBP/Cr)、蛋白肌酐比值(urinary protein to creatinine ratio,PRO/Cr),足细胞顶端膜性蛋白肌酐比值(Podocalyxin to creatinine ratio,PCX/Cr)、白蛋白肌酐比值(albumin to creatinine ratio,Alb/Cr)、免疫球蛋白G肌酐比值(immunoglobulin G to creatinine ratio,IgG/Cr)测定,然后对治疗前后结果进行分析评价。结果观察组氯沙坦治疗后尿PCX/Cr、Alb/Cr、PRO/Cr较治疗前有明显降低(P0.01),但尿β2-MG/Cr、RBP/Cr、IgG/Cr治疗前后比较无明显差异(P0.05);对照组尿PRO/Cr、ALB/Cr治疗后有所下降(P0.05),其余比值治疗前后无明显差异(P0.05)。观察组治疗后较对照组治疗后尿PCX/Cr、ALB/Cr、PRO/Cr有所降低(P0.05),β2-MG/Cr、RBP/Cr、IgG/Cr无明显差异(P0.05)。结论氯沙坦可明显减少IMN患者尿蛋白排泄,其机制可能与肾小球基底膜电荷屏障的修复有关,对肾小球基底膜的结构损伤无修复作用,也未发现对损伤肾小管明显的保护作用。     

头孢曲松泌尿系结石的理化分析

目的 探讨头孢曲松钠引发尿路结石的理化性质. 方法 2012年4月收检头孢曲松钠引发的儿童尿路结石2例.其中1例于2012年3月行头孢曲松钠治疗,过程中突发尿路结石,经治疗后自行排出沙粒状结石.另一例于2012年4月行头孢曲松钠治疗5d后突发下腹痛、呕吐,B超检查示右输尿管结石,KUB检查未见结石影,CT检查示右输尿管上段结石伴积水.保守治疗3d后结石排出,复查B超和IVU未见结石影.采用红外光谱、扫描电子显微镜及能谱仪分析2例尿路结石的成分. 结果 2例尿路结石样本均由头孢曲松和钙离子构成,摩尔比为1∶1. 结论 头孢曲松钠可引发尿石症,该结石具有X线下不显影的影像学特征,尿路结石成分为头孢曲松钙盐.     

Urinary creatinine excretion and protein/creatinine ratios vary by body size and gender in children

Urinary protein/creatinine ratio (Up/cr) is a simple measurement for evaluation of proteinuria. However, exact effects of body size and gender on urinary excretion of creatinine and Up/cr remain unknown. We aimed to clarify their effects. Early morning urine samples were collected from 124 children with urinary tract disorders. Urinary hourly excretion of creatinine, Ucr (in milligrams per hour), urinary hourly excretion of protein per body surface area, Up (milligrams per square meter per hour), and Up/cr (milligrams per milligram) were calculated. Effects of gender, age, body height, body weight and body surface area on Ucr and Up/cr were analyzed, respectively, in a multiple linear regression model. Body surface area and gender affected Ucr (r²=0.842, P<0.0001). Ucr adjusted by body surface area increased as body surface area grew with moderate variation. Up/cr showed a close correlation with Up and was affected by body height and gender as well. The regression equation showed that Up/cr values corresponding to the normal upper limit of Up, i.e., 4 mg/m²/h, in boys and girls 170 cm tall were approximately one third of those in children 80 cm tall (0.121 vs 0.043 for boys, 0.132 vs 0.047 for girls). The present study indicates that estimation of Up/cr needs to include consideration of children’s body height and gender.     

Rapid computer-assisted infrared analysis of urinary calculi using photoacoustic detection

The application of commercial spectrum-analysing software to quantitative analysis of urinary stones by Fourier transform infrared spectrophotometry is described. The infrared technique is straightforward in comparison with other stone analysis procedures of similar scope and affords significant time savings. The use of partial least squares regression in the analysis program enables better quantitation of stone components than has been hitherto possible using infrared methods. All the principal and many less common stone constituents can be detected and measured. Photoacoustic detection was employed, thus enabling non-destructive analysis with minimal sample preparation. A comparison is made between the infrared procedure, and the hybrid thermogravimetric plus wet chemistry technique, which it has superseded for routine urinary stone analysis in the authors' department.     

Dithio-bis-mercaptoethanesulphonate (DIMESNA) does not prevent cellular damage by metabolites of ifosfamide and cyclophosphamide in LLC-PK1 cells

Ifosfamide (IF) is an alkylating cytostatic with urotoxic (haemorrhagic cystitis) and nephrotoxic (Fanconi syndrome) side effects. Cyclophosphamide (CP), a structural isomer of IF, shows urotoxic but no nephrotoxic side effects. The development of haemorrhagic cystitis during therapy with IF or CP can be prevented by the uroprotective drug sodium-2-mercaptoethanesulphonate (MESNA). However, even in the presence of MESNA, Fanconi syndrome may still develop after therapy with IF. Using the renal tubular cell line LLC-PK₁, we investigated whether there is a protective effect of either MESNA or of its major metabolite DIMESNA, in combination with metabolites of IF or CP, on thymidine incorporation, uridine incorporation or total protein. DIMESNA, the dimer of MESNA, is the dominant form of the molecule in the circulation; the proximal tubular cell must convert this back to MESNA at the expense of glutathione, before it can exert its uroprotective action. We did not find a protective effect of DIMESNA under any of the experimental conditions tested. LLC-PK₁ cells exposed to 3 mmol/l DIMESNA did not convert DIMESNA to MESNA. The toxic effect of the CP metabolite 4-OOH-CP was more pronounced in the presence of DIMESNA than in its absence. MESNA completely prevented the toxic effects of acrolein and of 4-OOH-CP. The toxic effects of 4-OOH-IF and of chloracetaldehyde, two major metabolites of IF, were significantly reduced in the presence of MESNA. However, even at a 30-fold molar excess of MESNA over 4-OOH-IF, thymidine incorporation remained reduced by 40% compared with controls, indicating incomplete protection of tubular cells against metabolites of IF. Similarly, the effect of chloracetaldehyde was not completely reversed by MESNA. Our data indicate that DIMESNA is not metabolized sufficiently by LLC-PK₁ cells. Moreover, DIMESNA may deprive the cell of glutathione and thus enhance the toxicity of 4-OOH-CP. The finding that MESNA does not protect LLC-PK₁ cells completely against the toxic effects of IF metabolites may explain the difference in nephrotoxicity of IF and CP. It also explains the development of tubular damage in the presence of MESNA and in the absence of haemorrhagic cystitis.     

内外引流治疗肾损伤后尿外渗

目的：探讨肾外伤后尿外渗的诊断及治疗。方法：应用经皮肾周穿刺置管外引流及膀胱镜下置双J管内引流治疗肾外伤后尿外渗。结果：内、外引流治疗肾外伤后尿外渗3例，无肾周感染、复发等，效果满意。结果：内、外引流治疗肾外伤后尿外渗具有操作简单、损伤小、康复快等优点。     

Elemental analysis of urinary calculi by laser induced plasma spectroscopy

Laser induced plasma spectroscopy (LIPS) has been applied to analyse and identify elemental constituents of urinary calculi. Measurements on seven different urinary stone samples were conducted and the concentrations of some key elemental species were estimated. The elements detected with the present system were: Calcium, Magnesium, Sodium, Samarium, Potassium and Lead. Absolute concentrations of the species were derived from pre-calibration of the system for each element. Their concentrations were found to be widely different in different samples. It was observed that the samples containing a significant amount of lead have large proportion of calcium. It has been established that LIPS would allow real time clinic measurements of elemental contents and the concentrations in the biomaterials without sample preparation. The technique has the potential for routine clinic applications in urological disorder diagnosis.     

青少年尿路感染细菌分布以及耐药性分析

【摘要】 目的 探究近一年来深圳光明新区尿路感染青少年患者细菌分布情况和耐药情况。方法将2014年7月到2015年7月就诊光明新区医院的18~20岁尿路感染患者44例作为研究对象,收集中段尿分离培养细菌共得1382株菌落,分析尿路感染细菌的种类、分布和通过药敏试验分析耐药性。结果〓革兰氏阳性菌20.12%,真菌3.04%,革兰氏阴性菌76.85%;屎肠球菌37.41%,近平滑假丝酵母菌38.10%,大肠埃希菌50.47%。金黄色葡萄球菌对环丙沙星耐药性最强,屎肠球菌耐药性最强的为左氧氟沙星,粪肠球菌耐药性最强的为克林霉素,大肠埃希菌耐药性最强的为头孢曲松,铜绿假单胞菌耐药性最强的为氨曲南,肺炎克雷伯菌耐药性最强的为头孢曲松,柠檬酸杆菌耐药性最强的为环丙沙星。结论〓对于18~20的青少年岁的尿路感染患者,革兰氏阴性菌尤其是大肠埃希菌为主产生耐药性较为广泛。     

TVT和腹腔镜下膀胱颈悬吊术治疗女性单纯压力性尿失禁疗效的Meta分析

目的 系统评价经阴道无张力性尿道中段悬吊术(TVT)和腹腔镜下膀胱颈悬吊术(LB)治疗女性压力性尿失禁的客观成功率和并发症发生率.方法 计算机配合手工检索1994年至2006年6月PubMed、EMBASE及中国生物医学文献数据库和灰色文献中TVT、无张力性尿道悬吊术、膀胱颈悬吊术、压力性尿失禁等检索词.比较TVT和LB疗效的随机对照试验,采用Revman 4.2软件进行Meta分析,对TVT和LB术式的客观成功率、并发症发生率合并分析.结果 共纳入8个随机对照试验.TVT术式组259例,成功233例;LB组247例,成功189例.TVT组客观成功率高于LB组(RR为1.19,95%可信区间1.10～1.29,P<0.01),并发症发生率低于LB组(RR为0.48,95%可信区间0.32～0.71,P<0.05).结论 TVT术式客观成功率高于LB术式,而并发症低于LB术式.     

The role of Tamm-Horsfall protein in the pathogenesis of urinary tract infection

Although Tamm-Horsfall protein (THP) has been implicated in a variety of pathologic conditions, its physiologic function still remains unclear. Several studies have shown conclusively that bacteria expressing mannose-sensitive (MS) fimbriae bind to THP and to THP-coated exfoliated uroepithelial cells. Therefore it has been suggested that THP may prevent MS-fimbriated organisms from colonizing the epithelial surface of the urinary tract. Whereas older clinical studies were often inconclusive, recent studies using more sensitive assays demonstrated reduced THP levels in a geriatric population, in children with urinary tract infections and in elderly women with bacteriuria. Although additional studies are required, available data support a protective role for THP as a non-immune antibacterial host factor against uropathogens. The clinical importance of in vitro studies showing interference of THP with bacterial recognition by leukocytes and down-regulation of cell-mediated immune responses is presently unclear.     

红外光谱分析在复发性结石成分与初次结石成分比较中的应用

目的 探讨红外光谱分析联合饮食指导在预防结石复发中的效果。方法 回顾性分析2017年1月至2017年6月在本院收治的泌尿系结石患者1136例的临床资料,根据患者初次结石手术后是否结合红外光谱分析结果采取饮食指导分成观察组和对照组。观察组在初次结石手术后进行红外光谱分析结石成分并根据分析结果采取对应饮食指导的患者703例,对照组在初次结石手术后进行红外光谱分析结石成分但未采取饮食指导的患者433例。比较两组患者结石复发率及复发结石与初发结石成分差异。结果 观察组1年内结石复发患者85例（12.09%）,再次手术治疗42例（5.97%）;对照组1年内结石复发患者96例（22.17%）,再次手术患者68例（15.07%）;两组患者在结石复发和复发结石再次手术比较差异均有统计学意义（P<0.05）。观察组中42例患者复发结石符合手术指征,其中14例患者结石的主要成分发生了改变,而对照组中68例患者复发结石符合手术指征,其中7例患者的结石主要成分发生了改变。将两组患者中结石成分改变的比例进行比较,观察组结石主要成分发生变化的比例更高（P=0.005）。结论 红外光谱分析能较为准确地分析出结石成分,根据红外光谱分析结果进行饮食调整有利于降低患者结石复发风险。     

急性肾损伤患者尿肝脏型脂肪酸结合蛋白的水平变化及其意义

目的探讨急性肾损伤(acute kidney injury,AKI)患者尿肝脏型脂肪酸结合蛋白(livertype fatty acid binding protein,L-FABP)的水平变化及其意义。方法选择2013年10月至2015年12月我院收治的重症住院患者83例,根据患者入院时是否发生AKI分为AKI组和非AKI组,比较2组患者的各项生化指标[尿素氮、血肌酐、尿N-7乙酰-β-D葡萄糖苷酶(N-acetyl-β-D-glucosaminidase,NAG)、胱抑素C(cystatinC,Cys C)、肾小球滤过率]、L-FABP水平的差异,分析尿L-FABP水平与上述各生化指标的相关性以及尿L-FABP水平预测患者发生AKI的ROC曲线分析。结果 AKI组入院后第一次检查的血尿素氮、血肌酐、尿NAG、Cys C均高于非AKI组,差异有统计学意义(P0.05);AKI组的肾小球滤过率低于非AKI组,差异有统计学意义(P0.05);AKI组在入院12 h时尿L-FABP水平开始升高,而非AKI组的尿L-FABP水平维持在较稳定的水平;AKI组入院第12 h、第24 h、第48 h时尿LFABP水平均高于非AKI组,差异有统计学意义(P0.05);AKI患者入院第12 h的尿L-FABP与入院后第一次检查血尿素氮、血肌酐、尿NAG、Cys C均呈正相关(r=0.234,0.308,0.274,0.295),尿LFABP与肾小球滤过率呈负相关(r=-0.485);ROC曲线分析结果显示,诊断阈值为49.46,灵敏度为60.0%,特异度为95.83%,约登指数为0.558,曲线下面积为80.89%。结论 AKI患者的尿L-FABP水平异常升高,尿L-FABP可作为预测患者发生AKI的重要临床指标。     

Diagnostic value of urinary retinol-binding protein in childhood nephrotic syndrome

Urinary excretion of N-acetyl-β-d-glucosaminidase (NAG) and retinol-binding protein (RBP), sensitive markers of renal tubular damage and dysfunction respectively, were evaluated in paired remission and relapse urine samples from 16 patients (median age 12 years), with minimal change nephrotic syndrome (MCNS), in single samples from 5 nephrotic patients (median age 12 years) with focal segmental glomerulosclerosis (FSGS) and in 183 normal controls aged 2–16 years. The NAG and RBP data were expressed as a ratio over urinary creatinine (Cr). The NAG/Cr and RBP/Cr geometric means (ranges) for normal subjects were 11.1 (3.4–35.5) μmol 2-methoxy-4-(2"-nitrovinyl)-phenol (MNP)/h per mmol and 3.1 (0.3–38.8) μg/mmol, respectively. The NAG/Cr data revealed a weak negative correlation with age in normal children, whereas RBP/Cr was independent of age. RBP/Cr and NAG/Cr in MCNS in remission were the same as in controls. In MCNS in relapse, NAG/Cr was significantly elevated (P=<0.001), while in FSGS both RBP/Cr and NAG/Cr were significantly raised (P=<0.001 and P<0.008, respectively). These findings suggest that elevated NAG/Cr may be an indicator of relapse in both MCNS and FSGS and elevated RBP/Cr may allow differentiation between the two. Received May 7, 1997; received in revised form January 30, 1998; accepted February 4, 1998     

小儿多发性上尿路结石梗阻致急性肾功能衰竭的诊断及早期处理

目的：提高小儿上尿路多发性结石的诊断及早期治疗水平。方法：回顾性分析32例小儿上尿路多发性结石致急性肾功能衰竭的诊断及早期治疗过程，总结出较好的早期诊断与治疗的方法。结果：32例患者经腹部平片(KUB)及B超检查确诊27例，5例另行CT检查而确诊，32例患者均急诊处理，其中5例行输尿管镜碎石、4例经小儿膀胱镜置人小儿双J管而解除梗阻，23例经小儿膀胱镜置小儿双J管失败后，经常规手术解除梗阻，术后肾功能恢复良好。结论：小儿上尿路多发性结石致急性肾功能衰竭须早期诊断及治疗，KUB及B超检色是较好的诊断方法，应以简单、有效、创伤小的方法解除梗阻，可光用内镜方法治疗，如失败，则应急行常规手术解除梗阻。结石可留作Ⅱ期处理。     

Value of urinary L-FABP and NGAL in the diagnosis of acute kidney injury caused by obstructive nephropathy and the prediction of renal outcome

Objective To evaluate the values of urinary liver-fatty acid binding protein (uL-FABP) and urinary neutrophil gelatinase-associated lipocalin (uNGAL) in diagnosis of acute kidney injury (AKI) caused by obstructive nephropathy and in the prediction of renal prognosis. Methods Clinical data of 30 patients with obstructive nephropathy were collected prospectively. uL-FABP and uNGAL were measured by ELISA at various time points. Risk factors of the renal outcome were evaluated. The patients were followed up for at least one year. Results Patients with AKI had higher levels of uL-FABP and uNGAL compared to those without AKI [700.00(154.62-1216.14) μg/g•Cr vs 26.90 (16.77-41.38) μg/g•Cr; 1266.69 (671.57-3396.07) μg/g•Cr vs 179.12 (90.98-215.16) μg/g•Cr, all P＜0.01]. Positive correlations of uL-FABP and uNGAL with serum creatinine were found (r＝0.552, 0.553, all P＜0.01). The AUCs of uL-FABP and uNGAL to detect AKI were 0.925 and 0.900. Patients with non complete renal recovery had higher levels of uL-FABP before operation and 72-hour after operation compared to those with complete renal recovery (all P＜0.01). Before operation, the AUC of uL-FABP to detect renal prognosis was 0.948, sensitivity was 85.7% and specificity was 90.9%. 72-hour after operation, the AUC of uL-FABP to detect renal prognosis was 0.935, sensitivity was 85.7% and specificity was 90.9%. Kaplan-Meier analysis revealed that uL-FABP before operation over 366.57 μg/g•Cr or uL-FABP 72-hour after operation over 223.60 μg/g•Cr were closely related to the poor progression of renal function. Conclusions uL-FABP and uL-NGAL have good accuracy in detecting AKI. The level of uL-FABP before operation and 72-hour after operation is helpful to predict the renal outcome of obstructive nephropathy.