尿凝血酶原片段1及其mRNA在人肾组织中的表达

目的　了解人肾脏能否合成和分泌尿凝血酶原片段 1 (UPTF1 ) ,以及可能的合成和分泌部位。方法　收集 1 2例正常肾组织和 4例结石肾组织 ,采用免疫组织化学和逆转录 聚合酶链反应 (RT PCR)方法 ,研究UPTF1及其mRNA在人肾组织中的表达。结果　所有肾组织远曲小管和亨利氏袢升枝粗段的上皮细胞胞浆强阳性表达UPTF1 ,近曲小管上皮细胞浆染色为弱阳性 ,肾小球及间质组织未染色 ,正常人和结石患者肾组织的染色部位及强度差异无显著性 (P >0 .0 5) ;7例肾组织中 4例明显表达UPTF1的mRNA。结论　UPTF1来源于人肾脏的合成和分泌 ,可能是一种保护自身免于结石形成的生理性抑制因子。     

三叶因子1在大鼠草酸钙结石模型肾组织中的表达及其意义

目的 探讨三叶因子1(TFF1)在大鼠草酸钙结石模型肾组织中的表达及其与肾脏草酸钙结石形成机制的关系.方法 分别以1％的乙二醇溶液自由饮用和2％的NH4Cl溶液2 mL/d灌胃2周和4周;用偏光显微镜观察结石结晶形成情况;采用免疫组织化学染色法分别检测成石2周(n=20)、成石4周(n=20)和正常组(n=20)大鼠肾组织TFF1蛋白的表达.结果 TFF1免疫反应阳性物质主要位于肾小球、肾小管,与正常组比较,成石2周肾组织TFF1平均灰度值略下降(P＞0.05),成石4周平均灰度值明显降低(P＜0.05).结论 乙二醇诱导的大鼠草酸钙结石模型肾组织中TFF1低表达,伴随结石结晶数量和密度的增加,TFF1表达降低.TFF1在肾组织中的表达与乙二醇和NH4Cl干预的时限呈负相关;TFF1可能在肾草酸钙结石的形成中起抑制作用.     

过氧化物酶体增殖激活物受体-γ在肾细胞癌中的表达及其意义

目的观察过氧化物酶体增殖激活物受体(PPAR)-γ在肾细胞癌中的表达,探讨其意义。方法应用逆转录聚合酶链反应(RT-PCR)、免疫组织化学和Western blot从RNA及蛋白水平检测正常肾组织、肾癌组织和肾细胞株HK-2、HMCC;肾癌细胞株786-O、A498中PPAR-γ表达。结果肾癌组织、肾组织及细胞株中PPAR-γ mRNA普遍表达,肾癌中PPAR-γ蛋白表达量为正常肾的7.0~10.7倍;肾癌组织PPAR-γ阳性率为98.3％,肾组织中为60.0％,其表达强度差异有显著性(t=7.888,P<0.01)。PPAR-γ在肾癌的细胞核和细胞质中均表达;PPAR-γ表达和肾癌的分级、分期明显相关。结论核内受体PPAR-γ在人肾癌组织及肾癌细胞株中呈上调表达,提示PPAR-γ基因的激活可能是肾细胞癌治疗的一种新的方法。     

CyclinD1在肾细胞癌中的表达及其意义

目的探讨细胞周期蛋白D1（CyclinD1）在肾细胞癌中的表达及其意义。方法采用免疫组织化学技术,检测60例。肾细胞癌患者中肾癌组织及癌旁肾组织中CyclinD1的表达,并对Cyclin D1的表达与临床病理参数的关系进行分析。结果CyclinD1在肾细胞癌组织中的阳性率为83．3％,而癌旁肾组织中的阳性率为6．6％,两者具有显著性差异（P〈0．05）。Cyclin D1在肾细胞癌组织中的表达在不同性别、年龄、肿瘤体积组无显著差异（P〉0．05）。结论CyclinD1可作为判断肾细胞癌分化程度的标志物。     

慢性缺氧对大鼠肾组织内皮素-1及其受体mRNA的作用

缺氧可不同程度地影响人体正常的生理功能。有研究报道急性缺氧可引起肾脏血管收缩 ,肾功能减退 ,ET系统对此起重要作用[1] 。但慢性缺氧对肾功能的影响 ,以及ET起什么样作用尚不清楚 ,为此 ,本文建立了慢性缺氧大鼠模型 ,进行观察研究。材料与方法1　实验动物及模型复制　雄性SD大鼠 (温州医学院实验动物中心提供 ) 2 0只 ,体重 180～ 2 2 0g ,适应性饲养1周 ,然后随机分为两组。 (1)正常对照组 ,10只 ,作为正常氧浓度下的对照 ;(2 )缺氧组 ,10只 ,置于低压氧舱内 ,舱内充氮气 ,(室内温度 15℃～ 2 0℃ ,相对湿度 5 0 %～ 70 % )饲养 ,…     

狼疮肾炎肾组织中FcγR Ⅱb的表达及其意义

FcγRⅡb为免疫球蛋白超家旅成员，是IgG的低亲和力受体。我们探讨FcγRⅡb在狼疮肾炎(LN)的发病机制中的作用及其与LN活动性的关系。     

霉酚酸酯对糖尿病大鼠肾间质细胞浸润及肾组织ICAM-1和MCP-1 mRNA表达的影响

炎症反应是糖尿病肾病（DN）发生的重要环节，细胞间黏附分子（ICAM）和单核细胞趋化因子（MCP）表达增加是导致单核细胞及巨噬细胞浸润肾组织的两个重要因素。近年来发现霉酚酸酯（MMF）有抗炎作用，所以在理论上可以用于包含糖尿病肾病在内的一些非移植性疾病的治疗。本研究的目的主要在于探讨MMF对糖尿病大鼠肾脏ICAM-1和MCP-1m RNA表达的影响及其对肾脏的保护作用。     

载脂蛋白H在肾病综合征患儿肾组织表达的研究

目的研究载脂蛋白H(ApoH)在原发性肾病综合征(PNS)患儿肾组织中的分布及表达变化,探讨其在PNS进展过程中的作用。方法利用荧光定量RT-PCR和免疫组化检测78例不同病理类型的PNS患儿肾组织ApoH的mRNA及蛋白表达,并对肾组织中的肾小管损伤、间质炎症细胞浸润进行评分。肾活检前检测血白蛋白(Alb)、血脂、Scr、尿视黄醇结合蛋白(RBP)、尿蛋白定量。14例正常肾组织为对照。结果(1)正常和PNS患儿肾组织中均有ApoH蛋白表达,主要分布于皮质近肾小球的近端肾小管上皮细胞;正常及PNS患儿肾组织均有ApoHmRNA表达,且肾组织ApoH蛋白与mRNA表达呈显著正相关,r=0.264,P<0.05,提示肾脏本身能够合成ApoH.(2)不同病理类型PNS患儿肾组织ApoHmRNA及蛋白表达存在差异,微小病变NS(MCNS)、膜性肾病(MN)组ApoHmRNA及蛋白表达水平分别为4.95±0.40、4.73±0.60和12.06±2.04、12.35±0.61,与对照组(5.44±1.56和12.69±1.89)相比虽有减少,但差异无统计学意义,P>0.05;系膜增生性肾小球肾炎(MsPGN)、局灶性节段性肾小球坏死(FSGS)组ApoHmRNA及蛋白表达水平分别为3.30±0.28、2.82±0.36和10.13±3.09、10.12±1.02,明显低于MCNS、MN及对照组,差异均有统计学意义,P<0.05;(3)激素耐药组ApoHmRNA表达低于激素敏感组,分别为4.27±0.30、     

膀胱移行细胞癌组织中环氧化酶-2的表达及其意义

目的　探讨环氧化酶 2 (COX 2 )在膀胱移行细胞癌组织中的表达及其临床意义。方法应用逆转录 聚合酶链反应 (RT PCR)检测 5 2例膀胱移行细胞癌患者的癌组织及 17例患者距离肿瘤组织 3cm以外的正常膀胱黏膜组织中COX 2mRNA的表达 ,应用Westernblot法检测其中 4 9例膀胱癌组织及全部 17例正常膀胱黏膜组织中COX 2蛋白的表达 ,并应用条带密度分析软件半定量分析Westernblot条带密度。结果　COX 2mRNA在 83% (43/ 5 2 )的癌组织中表达 ,仅在 2 9% (5 / 17)的正常膀胱黏膜组织中表达 (P <0 .0 0 1) ;4 9例膀胱癌组织COX 2蛋白的表达量与肿瘤的分级、分期相关 (P<0 .0 0 1)。结论　COX 2在膀胱移行细胞癌组织中高表达 ,可能在肿瘤的发生发展中起一定的作用 ,可能成为膀胱癌化学预防的一个靶点。     

梓醇对5/6肾切除大鼠肾组织TGF-β1、CTGF表达的影响

目的:观察梓醇(catalpol)对肾衰大鼠肾组织TGF-β1、CTGF表达的影响。方法:将Wistar大鼠50只随机分为正常组,模型组,梓醇高、中、低剂量(100、50、10 mg/kg)组。除正常组外,其余各组大鼠均行5/6(ablation/infarction,A/I)肾切除法建立肾衰竭大鼠模型,从手术当日开始,梓醇各剂量组连续灌胃给药60 d。检测血清肌酐(Cr)、尿素氮(BUN)、超敏反应蛋白(hs-CRP)、血红蛋白(Hb)水平变化,ELISA法检测肾纤维化指标肾皮质转化生长因子(TGF-β1)、结缔组织生长因子(CTGF)的水平变化,HE染色观察大鼠肾脏病理变化,免疫组化法检测肾组织TGF-β1、CTGF的表达。结果:模型组大鼠血清Cr、BUN、hs-CRP水平显著升高(P<0.01)、Hb水平明显降低(P<0.01);血TGF-β1、CTGF含量显著上升(P<0.01);肾组织TGF-β1、CTGF的表达量增多。比较模型组,梓醇高(P<0.01)、中(P<0.05)、低(P<0.05)剂量组大鼠血清Cr、BUN、hs-CRP、TGF-β1、CTGF水平降低,Hb水平升高,肾组织TGF-β1、CTGF的表达量减少,其中尤以梓醇高剂量组疗效最佳。结论:梓醇可有效阻断大鼠肾纤维化进程、改善微炎症反应,保护大鼠肾功能,其作用机制可能与通过下调TGF-β1、CTGF表达,抑制纤维细胞增殖,减少纤维组织增生、炎性浸润相关,且具剂量依赖性。     

Effects of urinary prothrombin fragment 1 in the formation of calcium oxalate calculus

PURPOSE: We investigated the effects of urinary prothrombin fragment 1 in the formation of calcium oxalate urolithiasis. MATERIALS AND METHODS: Fresh urine and renal parenchyma from patients with calcium oxalate calculus and normal controls were collected. Urinary prothrombin fragment 1 was isolated and purified from urine. It was identified by sodium dodecyl sulfide-polyacrylamide gel electrophoresis and analysis of its first 13 N-amino acids. The inhibitory activity of urinary prothrombin fragment 1 on calcium oxalate crystal growth was tested by the seeded crystallization technique. Meanwhile, the gamma-carboxyglutamic acid composition of urinary prothrombin fragment 1 was analyzed by a previously described method and genetic mutation of the gamma-carboxyglutamic acid domain of urinary prothrombin fragment 1 from renal parenchyma was detected by polymerase chain reaction-single strand conformational polymorphism sequencing. RESULTS: The gamma-carboxyglutamic acid composition of urinary prothrombin fragment 1 was significantly decreased from normal (24.4 to 1.7 mol/1,000 amino acids) in patients with calcium oxalate calculus. The mean growth index +/- SD of urinary prothrombin fragment 1 to calcium oxalate crystals was 42.3 +/- 4.2 compared with the normal index of 19.2 +/- 2.8 (p <0.01). The polymerase chain reaction-single strand conformational polymorphism sequencing technique revealed no genetic mutation of the gamma-carboxyglutamic acid domain of urinary prothrombin fragment 1 in patients with calcium oxalate calculus. CONCLUSIONS: The gamma-carboxyglutamic acid composition of urinary prothrombin fragment 1 as well as its ability to inhibit calcium oxalate crystal growth was significantly decreased in patients with calcium oxalate calculus. This was not caused by genetic mutation of the gamma-carboxyglutamic acid domain of urinary prothrombin fragment 1. It is important to elucidate the mechanisms of calcium oxalate stones in view of urinary prothrombin fragment 1.     

A study of crystal matrix extract and urinary prothrombin fragment 1 from a stone-prone and stone-free population

South African blacks are immune to urinary calculi whereas whites have an incidence rate similar to that reported in Western societies. Urinary prothrombin fragment 1 (UPTF1) and the crystal matrix extract (CME) from which it is derived have been shown to be potent inhibitors of crystal growth and aggregation in undiluted human urine. The objective of the present study was to isolate CME and UPTF1 from the urines of black and white subjects in order to assess whether either might contribute to the black population's relative stone immunity. CME was isolated from freshly precipitated calcium oxalate (CaOx) crystals and a crystallization study was conducted in synthetic urine. Coulter Counter, ¹⁴C-oxalate deposition, and scanning electron microscopy data demonstrated that the extracts from both race groups strongly inhibited CaOx nucleation. The extract derived from the black subjects inhibited nucleation to a greater extent than that from the whites. A phase conversion from COM to COD in the presence of the extracts, in support of the inhibitory effect of CME, was also observed. Purified UPTF1 isolated from both groups' CME was subjected to rigorous biochemical characterization involving matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry, protein sequencing by Edman degradation, and amino acid analyses. No differences in molecular weight or amino acid sequence and composition were identified. It is suggested that the more potent inhibitory activity of the extract derived from the black subjects might be related to this group's relative stone immunity. Received: 8 August 2000 / Accepted: 16 November 2000     

无肾功能损伤大鼠肾草酸钙结石模型的筛选

目的 用最小的肾脏功能损伤筛选制作短时、经济和成石效果好的大鼠肾草酸钙结石模型.方法 将雄性SD大鼠90只随机分为两组.实验1采用1.0％氯化铵和不同浓度乙二醇(EC,0.1％、0.2％、0.4％、0.8％)处理.实验2单用不同浓度EG(0.1％、0.2％、0.4％、0.8％)处理.7～21 d后处死大鼠,测体质量、血、尿肌酐和尿草酸盐含量.苏木素-伊红(HE)染色观察肾组织病理改变及肾成石.结果 实验1中,不同浓度EG+ 1.0％氯化铵处理7d后,大鼠尿草酸盐排泄量均高于对照组(P＜0.05).用0.4％或0.8％ EG+ 1.0％氯化铵处理14d后,大鼠肌酐清除率均低于对照组(P＜0.01);光镜观察,肾皮质、髓质和肾乳头等处出现草酸钙结晶体,肾组织炎性损伤加重.实验2中,用0.4％或0.8％ EG处理7d后,尿草酸盐排泄量高于对照组(P＜0.01),但肌酐清除率变化差异无统计学意义(P＞0.05).光镜观察,肾皮质及间质未出现明显水肿,可观察到少量结石结晶但无炎性细胞浸润.结论 单用EG处理大鼠可以在没有严重肾损伤的情况下构造短时、经济和成石效果好的大鼠肾草酸钙结石模型.     

在尿路结石患者中预测草酸钙结石列线图的建立与应用

目的建立和应用个性化的列线图模型,探讨列线图预测尿路结石患者中草酸钙结石的准确性及可行性。方法回顾性分析2017年1月1日至2018年12月31日在中山大学附属第五医院接受手术治疗的298例泌尿系结石患者资料,以7∶3的比例随机分为建模组和验证组,基于建模组采用最小绝对值收敛和选择算子回归(Lasso)模型及多变量Logistic回归分析选择草酸钙结石的最佳预测特征,根据最佳预测特征以列线图的形式构建预测模型。通过C指数、校准曲线和决策曲线分别评估列线图的辨别力、校准和临床实用性,并基于验证组对外部验证进行评估。结果在LASSO模型中选择的最佳预测特征包括结石位置、甘油三酯(TG)和尿比重(SG)。将以上最佳预测特征和性别、年龄一起建立列线图模型后,建模组和验证组的C指数分别为0.706、 0.603,表明模型具有良好的辨别能力。校准曲线中标准曲线与预测校准曲线贴合良好,提示校正效果良好。决策曲线分析表明,在草酸钙结石可能性阈值为31%时使用该列线图可以在临床上获益。结论本研究建立的列线图预测模型可有效预测草酸钙结石,有助于筛选和早期识别草酸钙尿路结石的高危患者,对泌尿科医师进行临床治...     

肾结石大鼠肾组织bikunin mRNA的表达

目的 :研究bikunin在实验性肾草酸钙结石大鼠肾组织的表达及意义。方法 :采用乙二醇和氯化铵诱导大鼠肾草酸钙结石模型形成 ,检测各组大鼠肾功能、肾组织Ca2 + 含量和草酸钙晶体沉积、尿生化指标 ,并用逆转录聚合酶链反应 (RT PCR)检测bikuninmRNA在肾组织的表达情况。结果 :模型组大鼠的血清Cr、BUN、肾Ca2 + 含量、2 4h尿Ca2 + 、草酸 (Ox)分泌量和肾组织bikuninmRNA的表达均明显高于正常组 (P <0 .0 5 )。结论 :高草酸尿和草酸钙结晶的沉积能促使大鼠肾脏通过合成更多的bikunin来抑制大鼠肾组织草酸钙晶体的形成。     

Proteolysis and partial dissolution of calcium oxalate: a comparative, morphological study of urinary crystals from black and white subjects

Crystal adherence to the renal epithelium is widely regarded as a probable mechanism of stone formation. Intracrystalline proteins may provide access to the core of urinary crystals and thereby facilitate the dismantling of these crystals after their attachment to and phagocytosis by renal epithelial cells. The present study investigated the role of proteolysis and limited dissolution of urinary calcium oxalate (CaOx) crystals in South Africas white and black populations with a view to understanding the remarkably low stone incidence in the black population compared with the whites. CaOx crystals were precipitated from filtered urine or ultrafiltered urine dosed with an intracrystalline protein, urinary prothrombin fragment 1 (UPTF1), isolated from white and black subjects. The crystals were fractured and subjected to proteolysis and/or limited dissolution before examination using field emission scanning electron microscopy (FESEM). FESEM data showed that CaOx crystals from white and black subjects were eroded by treatment with proteases. Cathepsin D caused the most significant crystal erosion, and more noticeable degradation of CaOx monohydrate (COM) crystals compared to CaOx dihydrate (COD). Limited dissolution studies showed the unique ultrastructures and fragmentation processes of COM and COD crystals. COM crystals appeared to be more susceptible to degradation and dissolution than CODs. Since COMs are predominant in blacks, compared with COD crystals from whites, it is speculated that the lower stone rate amongst South African blacks might be attributed partly to their more efficient destruction of retained COM crystals.     

尿石症预防饮食管理软件管理肾结石患者饮食摄入的应用价值

目的探讨计算机尿石症预防饮食管理软件管理肾结石患者饮食摄人的使用经验及有效性和实用性。方法37例肾结石患者分2组，第1组23例为软件使用组，用尿石症预防饮食管理软件管理饮食摄人；第2组14例为常规饮食指导组，只接受医生对患者的常规尿石症预防饮食指导。分别记录2组患者饮食管理前后尿量、动物蛋白、钙、草酸、嘌呤、钠和镁等尿石症相关成分每日平均摄入量的变化。结果软件使用组患者饮食管理前后尿量从（1121±171）ml增至（1703±245）ml;动物蛋白食物可食部分的摄入从（180±26）g减至（146±13）g；钙摄入从（657±124）mg减至（912±99）mg；富含草酸食物可食部分摄入从（33±8）g降至（16士6）g；富含嘌呤食物可食部分从（30±10）g降至（15±6）g；以上指标变化差异均有统计学意义（P＜0．01），而钠、镁的摄入无明显变化。常规饮食指导组患者饮食管理前后尿量、动物蛋白食物可食部分、钙、富含草酸食物可食部分、钠、镁等比较差异均无统计学意义（P＞0．05），仅富含嘌呤食物可食部分从（28±10）g降低到（15±5）g，差异有统计学意义（P＜0．01）。结论计算机尿石症预防饮食管理软件可帮助尿石症患者有效管理饮食，为患者提供全面系统的饮食防石信息。     

凝血酶原分子中γ-羧基谷氨酸对草酸钙结晶的抑制作用研究

目的：探讨凝血酶原(PT)分子中γ-羧基谷氨酸(Gla)在抑制草酸钙结晶形成中的作用。方法：采用加入不同浓度的吗啉和甲醛(1：10000)或(1：100)，体外化学修饰PT，将其分子内8个或2个γ-羧基谷氨酸残基修饰转变成γ-亚甲基谷氨酸残基(γ-MGlu)，草酸钙种晶抑制试验测定化学修饰后PT对草酸钙结晶抑制作用的变化。结果：PT蛋白转变为2γ-MGlu的PT后对草酸钙结晶的抑制活性从正常对照的14．2降低至128(10％)；PT蛋白转变为8γ-MGlu后其抑制活性降低至5．0(65％)。结论：凝血酶原分子内γ-羧基谷氨酸(Gla)在抑制草酸钙结晶形成中具有重要作用。     

Face‐specific binding of prothrombin fragment 1 and human serum albumin to inorganic and urinary calcium oxalate monohydrate crystals

OBJECTIVE

To compare the intracrystalline distributions of prothrombin fragment 1 (PTF1) and human serum albumin (HSA) within inorganic and urinary calcium oxalate (CaOx) monohydrate (COM) crystals and to determine whether binding of PTF1 can be explained by interactions between particular γ‐carboxyglutamic (Gla) residues and atomic arrays on individual faces of the COM crystal.

MATERIALS AND METHODS

COM crystals were precipitated from inorganic solutions and ultrafiltered urine containing fluorescent HSA or PTF1 at different relative concentrations and examined by fluorescence microscopy. Accelrys Materials Studio and Discovery Studio were used to model the binding of PTF1 to the top, side and apical faces of the COM crystal.

RESULTS

PTF1 alone always adsorbed predominantly to the COM apical surfaces, while HSA bound principally to the side faces under inorganic conditions, but to the apical faces in urine. In the presence of each other, both proteins competed for adsorption to the apical faces, with attachment of PTF1 dominating over that of HSA. Modelling showed that urinary PTF1 had equal theoretical bonding potential for all three COM surfaces.

CONCLUSIONS

(i) Anisotropic inclusion of HSA and PTF1 into urinary and inorganic COM crystals results from their preferential binding to specific COM faces; (ii) the binding preference of HSA differs under inorganic and urinary conditions; (iii) preferential binding of PTF1 to the apical faces of COM is more complex than can be explained by interactions between Gla groups and surface atomic arrays; (iv) future studies of interactions between urinary proteins and stone mineral crystal surfaces should be performed in urine.