局灶节段性肾小球硬化患者足细胞钙神经蛋白的表达

目的:观察成人局灶节段性肾小球硬化(focal segmental glomerulosclerosis,FSGS)患者足细胞钙神经蛋白的表达并探讨其意义。方法:选取临床表现为肾病综合征且经肾活检确诊的成人特发性FSGS63例,微小病变(minimal change disease,MCD)24例和肾移植供肾组织10例作为对照。免疫组化法检测肾组织钙神经蛋白A(CnA)异构体α、β、γ的表达。免疫荧光法行肾组织CnAα和synaptopodin双标记,并采用胶体金免疫电镜对肾小球CnAα的表达定位。两位病理医生分别盲法评分染色结果。结果:供肾组织肾小球CnAα、β、γ免疫组化染色均阴性,肾小管CnAα、β微弱表达,CnAγ阴性。26例FSGS患者肾小球足细胞CnAα表达上调,其阳性率为41.27%;而MCD患者肾小球CnAα阴性(P0.01);两者肾小球CnAα、β、γ染色均阴性。足细胞CnAα阳性患者肾小管损伤指标尿视黄醇结合蛋白[20.91(0.17～54.37)mg/L]、血清肌酐[(128.18±56.58)μmol/L]高于阴性患者(P0.05);两者间病理类型构成差异存在统计学意义(P0.01);而年龄、性别、尿蛋白、尿N-乙酰-β-葡萄糖苷酶、血清白蛋白及胆固醇无统计学差异。塌陷型、细胞型、顶部型、门周型FSGS中足细胞CnAα阳性病例分别为5例(83.33%)、10例(66.67%)、8例(61.54%)、3例(20%),14例经典型FSGS患者均阴性。多因素Logistic回归分析示血清肌酐(OR4.855,P0.01)、塌陷型(OR11.069,P0.05)为FSGS患者足细胞CnAα过表达的主要相关因素。69.23%足细胞CnAα阳性患者肾小球syanptopodin表达减弱且不连续。结论:本研究首次发现部分FSGS患者足细胞上CnAα表达增强,这可能参与了FSGS的发病。FSGS与MCD患者足细胞CnAα的表达差异,提示二者发病机制不同。对临床怀疑FSGS而未见明确节段病变的患者,足细胞CnAα阳性有助于诊断。     

微小病变肾病和局灶节段性肾小球硬化患者贫血的临床特征对比研究

目的 对比分析微小病变肾病（MCD）和局灶节段性肾小球硬化（FSGS）患者贫血临床特征的差异。方法 采用回顾性队列研究方法，收集2012年2月至2017年2月东部战区总医院国家肾脏疾病临床医学研究中心经肾活检确诊的MCD和FSGS队列患者，根据有无肾小球系膜增生将MCD患者分为足细胞病组（174例）和MCD组（77例），并与FSGS组（110例）进行对比，分析3组患者贫血发生率、临床表现、肾病综合征（NS）疗效及预后的差异，并分析基线贫血的影响因素。将足细胞病组中尿糖阴性且无肾小球球性硬化患者纳入足细胞病1组（113例），余纳入足细胞病2组（61例），分别与MCD组和FSGS组进行对比，分析足细胞病亚组患者贫血发生率及NS疗效的差异。结果 MCD组、足细胞病组、FSGS组的NS疗效组间差异均有统计学意义（P<0.001）。FSGS组肾脏终点事件率、基线贫血发生率（11.82%、23.64%）均显著高于MCD组（1.30%、7.79%）和足细胞病组（1.2%、10.34%），差异有统计学意义（P<0.05）。3组贫血持续时间、贫血程度、贫血性质、贫血疗效差异无统计学意义（P&...     

狼疮足细胞病:一种特殊类型的狼疮性肾炎

系统性红斑狼疮患者临床表现为肾病综合征,肾活检病理表现为肾小球轻微病变(MCD)、系膜增生(Ms P)或局灶节段性肾小球硬化(FSGS),超微结构以足细胞足突广泛融合为特征,毛细血管袢内皮下及上皮侧无免疫沉积物的肾损害称为狼疮足细胞病。肾小球病理改变为MCD和Ms P的狼疮足细胞病患者,血尿发生率低,激素治疗敏感,但单用激素维持的复发率高达90%,激素联合其他免疫抑制剂维持可显著降低复发率。而病理表现为FSGS的狼疮足细胞病患者,急性肾损伤的发生率高,肾小管间质损伤重,激素治疗缓解率低。狼疮足细胞病复发后可发生病理转型,远期预后良好。狼疮足细胞病应作为一类特殊类型狼疮性肾炎纳入新的病理分型体系中。     

狼疮性肾炎足细胞病患者的临床病理特征及预后

目的:回顾性分析狼疮性肾炎足细胞病(狼疮足细胞病)患者的临床、病理特征及远期预后。方法:系统性红斑狼疮(SLE)伴肾损害,经肾活检组织学和电镜检查符合狼疮足细胞病的患者53例(女48例,男5例,中位年龄31岁,中位病程1.5月)。回顾性分析其临床、病理特征及远期预后。结果:53例狼疮足细胞病占同期狼疮性肾炎的1.41%,其中50例临床表现肾病综合征,17例(32.1%)伴急性肾损伤(AKI),合并镜下血尿和高血压各9例(17.0%)。根据肾活检光镜改变分为系膜增生性病变(MP,n=31)、轻微病变(MCD,n=13)和局灶节段性肾小球硬化(FSGS,n=9)三组。电镜观察中位足细胞足突融合比例85%,三组间无明显差异。FSGS组AKI发生率(77.8%)显著高于MP组(22.6%)和MCD组(23.1%)(P0.01),肾小管间质急性病变程度也明显高于其他两组(P0.05)。与MCD组(23.1%)相比,MP组(83.9%)和FSGS组(88.9%)低C3血症的比例显著升高(P0.01)。经激素或激素联合免疫抑制剂诱导治疗后,69.8%获得完全缓解,FSGS组完全缓解率(22.2%)显著低于MCD组(92.3%)和MP组(74.2%)(P0.01)。中位随访时间60月,29例(54.7%)肾病复发,13例复发后行重复肾活检,其中6例发生病理转型,无终末期肾病或死亡病例。结论:狼疮足细胞病以肾病综合征或伴AKI为主要特征,组织学改变可为MCD、MP或FSGS,激素或激素联合免疫抑制剂治疗敏感,FSGS者AKI发生率高、肾小管损伤重且治疗缓解率低。该病复发率高,部分重复肾活检可见病理转型,长期随访预后良好。     

肥胖相关性肾病肾组织中nephrin desminWTI的表达

目的 研究肥胖相关性肾病患者肾小球足细胞病变,分析不同肾组织病理类型患者肾小球足细胞中neph-rin、desmin、WT1的表达和分布特征.方法 收集2001-2008年河北医科大学第二医院肾内科收治的16例经肾穿刺活检明确诊断的肥胖相关性肾病患者资料,根据其肾脏病理分为肥胖相关性肾小球肥大症(O-GM)组11例,肥胖相关性局灶节段性肾小球硬化症(O-FSGS)组5例.对照组5例,为外科切除的且已诊断为肥胖的肾脏肿瘤标本的正常皮质组织.应用免疫组织化学方法检测足细胞特异标志物nephrin、desmin、WT1,同时结合临床及肾组织病理有关指标的进行多因素分析.结果 nephrin、desmin、WT1在正常肾组织沿肾小球基底膜呈连续、线形分布.O-GM组nephrin、desmin、WT1的表达量降低,与对照组比较,差异无统计学意义,O-FSGS组nephrin、desmin、WT1的表达下降,呈点状、短线条状,较正常组织少,差异有统计学意义(P<0.05).O-FSGS组足细胞计数少于O-GM组,差异有统计学意义(P<0.01),且足细胞融合、微绒毛化改变和肾小球硬化数明显增多(P<0.05).结论 肥胖相关性肾病患者早期即出现肾小球足细胞中nephrin、desmin、WT1的表达和分布减少,随着病变的加重,这种变化愈发明显.     

Fabry病的临床表现及肾脏病理学特征

目的 :探讨Fabry病患者临床表现及肾组织形态学特点。　 方法 :总结 9例Fabry病患者临床表现和肾组织形态学特点。同时对肾小球足细胞数定量分析 ,并借助足细胞特殊标记物WT1对足细胞的密度及其缺失分布特点进行观察。　 结果 :①患者以年轻男性为主 ,多有肾脏病家族史。②多伴有Fabry病肾外表现 ,表现为皮肤血管角质瘤、少汗、低热、肢端感觉异常等症状。心脏受累发生率较高 ,多表现为房室传导阻滞、PR间期缩短、ST段和T波的异常、左心室高电压等。③均无明显角膜混浊、视网膜动脉曲张等Fabry病常见眼部病变。④肾脏病变表现为轻至中度蛋白尿。部分患者伴轻微镜下血尿。肾小管间质损伤较突出。⑤肾组织形态学改变表现为光镜下肾小球足细胞弥漫空泡变性 ;甲苯胺蓝染色显示足细胞浆内含大量嗜甲苯胺蓝的蓝色颗粒状物 ;电镜下嗜锇性、同心圆样髓样小体在胞浆中堆积。多数患者小管间质中重度病变。肾间质血管病变现象较为突出。⑥肾小球足细胞计数及足细胞密度均明显低于正常 (P <0 0 1) ,WT1表达缺失呈节段分布 ,缺失部位多位于周边袢和有节段硬化病变处。　 结论 :①Fabry病以男性青年多见 ,多有肾脏损害的家族史。②多数患者存在肾外表现 ,心脏病变发生率较高。③无明显Fabry病特征性眼部病变 ,无明显     

足细胞研究进展

足细胞是肾小球滤过屏障重要组成部分。足细胞损伤是肾小球疾病发生发展的关键环节,是导致蛋白尿、肾小球硬化及肾功能进行性恶化的病理生理基础。局灶节段性肾小球硬化(FSGS)和微小病变(MCD)主要表现为肾小球足细胞病变。足细胞损伤机制和靶向治疗一直是肾小球疾病研究的热点,本文将综述足细胞基因组、转录组研究进展和新发现的足细胞损伤相关基因,重点阐释足细胞损伤机制研究进展。     

糖尿病肾病患者足细胞病变的临床病理特征

目的：研究2型糖尿病肾病患者肾小球足细胞病变，分析其与血糖控制、蛋白尿、肾功能及肾组织病理形态学改变之间的关系。方法：27例经肾穿刺活检明确诊断的2型糖尿病肾病。肾组织病理定量计数肾小球足细胞数。借助足细胞特定标志物WTl与肾小球基膜Ⅳ型胶原α3链(C—Ⅳα3)双套色荧光染色，用激光共聚焦荧光显微镜对肾小球足细胞进行准确的密度定量分析。结果：糖尿病肾病患者无论临床表现为微量白蛋白尿、蛋白尿(尿蛋白＜3．5g/24h)，还是大量蛋白尿(尿蛋白＞3．5g/24h)，均伴肾小球足细胞数目及密度的减少，其中以大量蛋白尿组最为显著，蛋白尿组次之，微量白蛋白尿组最轻。足细胞数目及其密度与尿蛋白量呈显著负相关(P＜0．01)。足细胞数目的减少还与肾小球病理改变相关，表现为肾小球K—W结节病变者其足细胞数明显少于肾小球弥漫系膜增生性病变者(P＜0．01)。此外，足细胞数减少严重者肾小球硬化以及血清肌酐水平升高的发生率均明显增高。结论：糖尿病肾病患者早期就表现出肾小球足细胞数目及其密度的减少，随着病变加重，这种变化愈发明显。足细胞病变不仅导致大量蛋白尿的发生，而且还与肾小球硬化和肾功能损伤的发生有关。     

KDIGO指南解读：成人微小病变肾病和特发性局灶节段性肾小球硬化治疗

微小病变肾病(MCD)和特发性局灶节段性肾小球硬化(FSGS)是成人肾病综合征的常见病理类型。2012年6月发表的改善全球肾脏病预后组织(KDIGO)指南对多种原发及继发性肾小球肾炎的治疗给出了最新意见,对初发、复发、激素依赖和激素抵抗型MCD和FSGS患者的激素使用剂量、疗程及一线免疫抑制剂的选择等给予了建议。本文结合中国患者的特点对该指南关于MCD和FSGS治疗的建议做一解读。     

糖尿病肾病患者尿足细胞脱落及其与尿蛋白的关系

目的 通过观察糖尿病肾病(DN)患者尿中足细胞及肾活检标本足细胞脱落情况,探讨尿中足细胞脱落与尿蛋白、空腹血糖、肌酐的关系.方法 选取2005年1月至2006年12月间天津医科大学总医院收治的DN患者50例,微小病变(MCD)患者25例及正常人对照组10例,按照慢性肾脏病(CKD)分期将DN患者分为5组,其中CKD I期15例,CKD Ⅱ期10例,CKD Ⅲ期8例,CKD IV期9例,CKD V期8例;另按尿蛋白量将DN患者分为3组,其中少量蛋白尿组(<1.0 g/d)13例,中量蛋白尿组(1-3.5 g/d)20例,大量蛋白尿组(>3.5 g/d)17例,采用间接免疫荧光法检测尿足细胞数目及肾小球足细胞特异性蛋白(podocalyxin,PCX)表达;分别统计各组中尿足细胞数目;同时检测各组患者24h尿蛋白、肌酐、空腹血糖,多组计量指标比较采用单因素方差分析,双变量分析采用Pearson相关分析.结果 DN组尿中足细胞的阳性率为88%(44/50);微小病变(MCD)组及正常对照组中阳性率为0;DN组患者的肾活检标本中,肾小球足细胞PCX表达缺失,MCD组肾活检标本足细胞PCX呈完整连续性表达.少量蛋白尿组尿足细胞数目(1.5±1.0)个/ml,中等量蛋白尿组(2.2±0.7)个/ml及大量蛋白尿组(3.5±1.3)个/ml,除少量蛋白尿与中量蛋白尿组差别不具有统计学意义外,其他组间差别均具有统计学意义(P<0.05).DN患者尿足细胞数目与蛋白尿(r=0.785,P<0.05)、I-Ⅲ期患者同期SCr呈正相关(r=0.466,P<0.05);而与FBG无相关性(P>0.05).结论 DN进展过程中存在足细胞损伤.DN足细胞脱落与尿蛋白、CKD I至Ⅲ期同期SCr呈正相关,与Ⅳ至V期SCr、FBG无关.     

1-Naphthyl isocyanate and 1-naphthylamine as metabolites of 1-naphthylisothiocyanate

Abstract: The importance of the bioactivation of 1-naphthylisothiocyanate was studied. Forty minutes after 1-naphthylisothiocyanate administration to rats, bile was collected over a 2.5-h period; the liver was then excised and homogenized. 1-naphthylisothiocyanate and its metabolites in bile and liver of rats were identified and quantified using coupled gas chromatography-mass spectrometry. Three main compounds were found in all 1-naphthylisothiocyanate-treated animals. They were identified as 1-naphthyl isocyanate, 1-naphthylamine and the parent compound, 1-naphthylisothiocyanate. When rats were given cycloheximide, which attenuates 1-naphthylisothiocyanate toxicity, 30 min before 1-naphthylisothiocyanate (300 mg/kg), 1-naphthyl isocyanate concentration was significantly lower than in rats receiving only 1-naphthylisothiocyanate. The appearance of 1-naphthylamine was also inhibited by cycloheximide, although not to the same extent as 1-naphthyl isocyanate. On the other hand, phenobarbital, which potentiates 1-naphthylisothiocyanate hepatotoxicity, enhanced 1-naphthyl isocyanate and 1-naphthylamine formation. It is suggested that 1-naphthyl isocyanate, 1-naphthylamine and the highly reactive sulfur released from 1-naphthylisothiocyanate might be involved in the hepatotoxic effect of 1-naphthylisothiocyanate.     

The target of ezetimibe is Niemann-Pick C1-Like 1 (NPC1L1)

Ezetimibe is a potent inhibitor of cholesterol absorption that has been approved for the treatment of hypercholesterolemia, but its molecular target has been elusive. Using a genetic approach, we recently identified Niemann-Pick C1-Like 1 (NPC1L1) as a critical mediator of cholesterol absorption and an essential component of the ezetimibe-sensitive pathway. To determine whether NPC1L1 is the direct molecular target of ezetimibe, we have developed a binding assay and shown that labeled ezetimibe glucuronide binds specifically to a single site in brush border membranes and to human embryonic kidney 293 cells expressing NPC1L1. Moreover, the binding affinities of ezetimibe and several key analogs to recombinant NPC1L1 are virtually identical to those observed for native enterocyte membranes. KD values of ezetimibe glucuronide for mouse, rat, rhesus monkey, and human NPC1L1 are 12,000, 540, 40, and 220 nM, respectively. Last, ezetimibe no longer binds to membranes from NPC1L1 knockout mice. These results unequivocally establish NPC1L1 as the direct target of ezetimibe and should facilitate efforts to identify the molecular mechanism of cholesterol transport.     

Inhibition of procarcinogen-bioactivating human CYP1A1, CYP1A2 and CYP1B1 enzymes by melatonin

Abstract:  Administration of melatonin to rodents decreases the incidence of tumorigenesis initiated by benzo[ a ]pyrene or 7,12-dimethylbenz[ a ]anthracene, which requires bioactivation by cytochrome P450 enzymes, such as CYP1A1, CYP1A2 and CYP1B1, to produce carcinogenic metabolites. The present study tested the hypothesis that melatonin is a modulator of human CYP1 catalytic activity and gene expression. As a comparison, we also investigated the effect of melatonin on the catalytic activity of CYP2A6, which is also a procarcinogen-bioactivating enzyme. Melatonin (3–300 μ m ) decreased 7-ethoxyresorufin O -dealkylation catalyzed by human hepatic microsomes and recombinant CYP1A1, CYP1A2 and CYP1B1, whereas it did not affect coumarin 7-hydroxylation catalyzed by hepatic microsomes or recombinant CYP2A6. Melatonin inhibited CYP1 enzymes by mixed inhibition, with apparent K _i values (mean ± S.E.M.) of 59 ± 1 (CYP1A1), 12 ± 1 (CYP1A2), 14 ± 2 (CYP1B1) and 46 ± 8 μ m (hepatic microsomes). Additional experiments indicated that melatonin decreased benzo[ a ]pyrene hydroxylation catalyzed by hepatic microsomes and CYP1A2 but not by CYP1A1 or CYP1B1. Treatment of MCF-10A human mammary epithelial cells with melatonin (up to 300 μ m ) did not affect basal or benzo[ a ]pyrene-inducible CYP1A1 or CYP1B1 gene expression. Consistent with this finding, melatonin did not influence reporter activity in aryl hydrocarbon receptor-dependent pGudluc6.1-transfected MCF-10A cells treated with or without benzo[ a ]pyrene, as assessed in an in vitro cell-based luciferase reporter gene assay. Overall, melatonin is an in vitro inhibitor of human CYP1 catalytic activity, and it may be useful to develop potent analogues of melatonin as potential cancer chemopreventive agents that block CYP1-mediated chemical carcinogenesis.     

甲型H1N1与季节性H1N1流感病毒血凝素基因比较分析

目的分析泰安市2008～2009年度季节性流感与2009年度甲型H1N1流感病原学检测结果 ,比较季节性H1N1与甲型H1N1血凝素基因变异情况。方法选择国家级流感监测哨点医院以及暴发疫情的疫点,采集流感样病例的鼻咽拭子标本,通过RealtimePCR进行病毒检测,用MDCK细胞进行病毒分离,通过RT-PCR扩增血凝素HA1片段的基因并测序,利用生物信息学进行序列分析。结果 2008～2009年共检测鼻咽拭子标本283份,分离出流感病毒33株,分离阳性率为11.67%,其中季节性H1N1亚型31株。2009年5月1日～12月31日,检测鼻咽拭子标本996份,流感核酸检测阳性417份,阳性率为41.86%,其中甲型H1N1337份,季节性H1N1亚型1份。6株季节性H1N1病毒均在多个氨基酸位点上发生变异,与疫苗株A/Brisbane/59/2007(H1N1)比较,有11个位点发生了突变,其中5个位点位于抗原决定簇上;测序成功的6株甲型H1N1病毒在多个氨基酸位点发生变异,与疫苗株A/California/07/2009(H1N1)比较,有6个位点发生突变,其中1个位点位于抗原决定簇的B区。结论 2008～2009年度季节性H1N1为优势株,甲流暴发后,甲型H1N1成为绝对优势毒株。季节性H1N1分离株有多处氨基酸替换,抗原决定簇B区变异频繁;甲型H1N1病毒分离株的基因有变异,但关键位点第222位仍为D(天冬氨酸),与疫苗株相比抗原决定簇的关键位点变化不大。     

Characterization of MTHFR,GSTM1, GSTT1, GSTP1, and CYP1A1 genotypes in childhood acute leukemia

The role of methylenetetrahydrofolate reductase (MTHFR C677T), glutathione S-transferases (GSTM1 and GSTT1 null, GSTP1 Ile105Val), and cytochromes p450 (CYP1A1*2A) genotypes in the etiology of childhood leukemia was simultaneously investigated. 144 Turkish children with acute lymphoblastic leukemia (ALL) and 33 with acute nonlymphoblastic leukemia (ANLL) were studied and compared with 185 healthy pediatric controls. The frequency of MTHFR genotype was insignificantly higher in ALL (7.7%) and ANLL (6.3%) than in controls (4.4%). Equal distribution of the GSTM1 null genotype was detected between ALL patients and controls (55%), while its incidence was slightly higher in ANLL patients (61.3%). Although GSTT1 null genotype was insignificantly lower in ALL patients (20.9%) than controls (22.7%), it was significantly underrepresented in ANLL patients (6.5%) (P = 0.05, OR 0.24, 95% CI 0.05-1.03). The homozygous frequency of GSTP1 genotype did not differ significantly between groups of ALL (3.7%), ANLL patients (9.1%) and controls (4.9%). Homozygous CYP1A1*2A genotype was underrepresented in ALL patients (1%) as compared to control (4.8%) but the differences did not reach to statistical significance (OR 0.21; 95% CI 0.03-1.72). Homozygosity for this genotype was not detected in ANLL patients. No particular association was noted between different combinations of combined genotypes and risk of development of childhood ALL and ANLL. These results suggested that there are no significant associations between the studied genotypes and the risk of developing either form of acute leukemia except GSTT1 null and homozygosity for CYP1A1 genotypes that may play protective roles in the development of ANLL in Turkish children.