成纤维细胞生长因子23在慢性肾脏疾病血磷代谢中的作用

 成纤维细胞生长因子23(FGF23)是近年来发现的由骨细胞产生的一种激素,主要作用是抑制肾小管磷重吸收和1,25(OH)₂D循环水平。慢性肾脏疾病(CKD)是世界范围内影响数百万人的公共卫生疾病。FGF23循环水平升高导致的血磷稳态失调是CKD早期和普遍的并发症。本文综述FGF23家族结构、FGF23在调节磷和维生素D代谢中的作用及其合成和分泌的调控。重点介绍FGF23在CKD中与血钙、血磷及甲状旁腺激素(PTH)的作用机制,以及FGF23在评估CKD进展、预测预后中的作用。分析表明,FGF23可能成为慢性肾脏疾病骨和矿物质代谢中的重要诊断标志和治疗对象。     

肾移植后伴骨质疏松症患者血清骨代谢指标的变化

应用放射免疫分析法和免疫放射分析法对７例已确诊有骨质疏松症的骨移植患者进行血清骨代谢指标检测。结果表明患者成骨细胞活性指标（血清特异性骨碱怀磷酸酶、骨钙素和Ｉ型前胶原蛋白羧基末端前肽）和破骨细胞功能指标（血清Ｉ型胶原交联羧基末端肽）明显高于正常对照组，患者血清甲状旁腺素和２５羟维生素Ｄ水平正常。提示肾移植后伴骨质疏松症患者的成骨细胞活性和破骨细胞活性明显增高，但无继发性甲状旁腺功能亢进和维生素Ｄ代     

5 /6 肾切除小鼠中矿物质与骨代谢紊乱的特征

摘要: 目的通过制备5 /6 肾切除( 5 /6 Nx) 小鼠慢性肾脏病( CKD) 模型,观察小鼠成纤维生长因子23( FGF23) 及钙磷代谢指标,有助于研究矿物质与骨代谢紊乱的发病机制。方法28 只C57 雄性小鼠适应性饲养1 周后,切除模型组约5 /6 左肾,1周后切除右肾, 16 周后处死后收集血尿和肾脏组织标本,常规检测血尿FGF23、甲状旁腺素( PTH) 、活性维生素D、钙、磷等指标,观察肾脏病理。结果与假手术组和正常对照组相比,术后16 周5 /6 Nx 小鼠出现体质量减低,血清FGF23、PTH,血磷等异常升高,活性维生素D 水平显著降低,尿蛋白增加。肾脏病理发现系膜基质增生、胶原沉积及纤维化明显,和肾小球代偿性肥大。结论5 /6 Nx 小鼠可以出现明显的矿物质与骨代谢紊乱特征,是研究CKD 骨病合适的模型。     

重组人甲状旁腺素1-34在毕赤酵母克隆及表达的研究

人甲状旁腺素(Parathyroid hormone,PTH)是由人甲状旁腺主细胞分泌的直链肽,含84个氨基酸,是维持体内钙磷平衡的主要激素。PTH通过识别骨骼、小肠和肾脏等细胞表面特异性受体,与降钙素和维生素D共同调节Ca2+,PO43-的代谢,具有明显的成骨作用。研究发现,PTH的N-端34肽(PTH 1-34)具有完全的PTH受体结合能力与生物活性。目前PTH 1-34作为临床治疗骨质疏松症的注射剂已在美国上市,但价格昂贵且需要每天注射给药。而国内关于PTH1-34的临床研究很少,所以有必要通过基因工程手段获取PTH 1-34,降低成本并可大规模制备目的蛋白。本文概述了PTH近年来在国内外的研究情况,利用目前发展较快的毕赤酵母真核表达系统,构建了高效表达PTH 1-34毕赤酵母工程菌。为今后上发酵罐摸清条件。     

短体副鳅泌尿系统的组织学研究

采用常规组织学方法对短体副鳅泌尿系统进行了研究,结果表明:短体副鳅泌尿系统由中肾、输尿管、膀胱构成,未观察到头肾.中肾包括肾小体、肾小管和填充于其间的拟淋巴组织,无皮质和髓质之分.肾小管由第一近端小管、第二近端小管和远端小管组成,无颈段.输尿管前后段组织结构有明显差异.膀胱为不发达的输尿管膀胱.斯坦尼斯小体多枚.     

纳米金免疫层析法定量检测尿RBP

应用纳米金免疫层析技术(双抗夹心法)和柠檬酸三钠还原法建立一种对尿RBP的快速定量检测方法,并在此基础上研制出快速检测试纸条.该试纸条用纳米金免疫层析定量分析仪可在15 min内实现RBP定量检测,检测限为150μg/L(相当于150 ng/mL),检测范围为150～5000μg/L.与尿微量白蛋白(ALB)、转铁蛋白(TRF)、β2-微球蛋白(β2-MG)、尿纤维连接蛋白(FN)、溶菌酶(LZM)等肾脏疾病标志物无交叉反应.该方法特异性强、检测灵敏度高、范围广、简便快捷,在近端肾小管的损伤、糖尿病肾病等肾脏疾病的早期诊断及过程监控中具有较为广泛的临床应用.     

Vasopressin stimulates formation of coated pits in rat kidney collecting ducts

The water permeability of collecting ducts is greatly increased by the antidiuretic hormone, vasopressin. This permeability increase is associated with the appearance of intramembrane particle (IMP) clusters on the luminal plasma membranes of principal cells of the collecting duct epithelium. IMP aggregates have also been related to an increase in water permeability of two other vasopressin-sensitive epithelia, the amphibian urinary bladder and the amphibian epidermis, and it has been proposed that these specialized membrane domains might represent specific water-permeable membrane patches, induced by the hormone in their respective epithelia. Using a cytochemical probe for membrane cholesterol, filipin, we show here that the membrane patches in rat kidney are selectively devoid of filipin-sterol complexes and that when identified in thin sections, they have a cytoplasmic bristle coat: both of these features are characteristics of coated pits which, in other systems, are involved in receptor-mediated endocytosis. We also show that vasopressin induces the appearance of coated pits on collecting duct luminal membranes in Brattleboro homozygous rats, which have hereditary diabetes insipidus.     

珍珠鸟和家鸡甲状旁腺激素3型受体(PTH3R)的结构与表达图谱分析

甲状旁腺激素(PTH)与动物钙稳态调控和骨代谢平衡相关,其生理作用是通过甲状旁腺激素受体(PTHR)介导。甲状旁腺激素受体家族包括三个不同的亚型,其中甲状旁腺激素3型受体(PTH3R)在非哺乳类脊椎动物生长发育过程中起着重要作用,然而PTH3R在鸟类中的研究则相对较少。 本研究采用RT-PCR方法,首先克隆了珍珠鸟和家鸡的PTH3R基因全长cDNA序列。结果显示,家鸡PTH3R (cPTH3R) cDNA全长1632 bp,编码543个氨基酸,珍珠鸟PTH3R(zPTH3R-w) cDNA序列全长1563 bp,编码520个氨基酸,其蛋白均含有信号肽序列、七次跨膜区等特征性结构。此外,在珍珠鸟中还发现一个新剪接变体zPTH3R-v1,其cDNA序列全长1468 bp,编码488个氨基酸,其缺失了第3外显子进而导致第1跨膜结构域缺失。利用生物信息学方法,我们还对珍珠鸟和家鸡PTH3R蛋白序列进行三维建模。 采用RT-PCR方法,本研究也对珍珠鸟PTH3R基因进行组织表达分析。结果显示,zPTH3R及其剪切变体zPTH3R-v1在珍珠鸟脑及外周组织中广泛表达。珍珠鸟和家鸡PTH3R基因的成功克隆与结构解析,将为下一步开展PTH3R在鸟类中的功能研究奠定重要基础。     

基于线性判别式和支撑向量机的肾结石分类方法

用支撑向量机(SVM)方法辅助诊断肾结石,并和线性判别式方法作比较,结果显示这两种方法都表现出了很好的预测能力.鉴于SVM是用于解决非线性的良好方法.因此,SVM作为一种有效的机器学习方法是可以用来进行肾结石的辅助诊断和分类研究的.肾结石的成因比较复杂,与自然环境、社会生活条件、全身性代谢紊乱及泌尿系统本身的疾患有关,本文从钙离子生物学特性方面讨论了钙盐结石的成因.     

双侧上尿路结石49例分析

本文报道双侧上尿路结石49例,占同期尿路结石的8.33%(49/588)。本组年龄以21～40岁最多见(29例)。总肾功能正常者29例,合并尿毒症者20例。体会对双侧上尿路结石应积极采取手术治疗;对急性梗阻严重肾功能障碍和(或)感染者,应积极解除梗阻或先作引流;而对梗阻造成慢性肾功能障辞者,应先进行非手术治疗,待肾功能好转后再行手术。肾部分切除术在总肾功能尚好的情况下,不失为一种较好的术式。对双侧上尿路结石的手术侧选择问题,根据肾对抗平衡原理,按下列原则处理:1.结石梗阻不严重,肾功能正常者:(1)双侧输尿管石,应先处理梗阻较重,症状较剧的一侧或双侧结石一次摘除。(2)一侧肾石,对侧输尿管石,一般先摘除输尿管石。(3)双侧肾石,应先处理病变较轻,手术比较简单、安全的一侧。2.结石梗阻和(或)感染较重,并发慢性肾功能不全者:原则上应双侧同期手术,并另置造瘘管遣瘘;若一侧感染严重,对侧结石亦难以取出者,则应先作感染侧引流,再作对侧手术,最后完成引流侧的取石手术。     

Na(+)/H(+ ) exchanger regulatory factor 2 directs parathyroid hormone 1 receptor signalling

The parathyroid hormone 1 receptor (PTH1R) is a class II G-protein-coupled receptor. PTH1R agonists include both PTH, a hormone that regulates blood calcium and phosphate, and PTH-related protein (PTHrP), a paracrine/autocrine factor that is essential for development, particularly of the skeleton. Adenylyl cyclase activation is thought to be responsible for most cellular responses to PTH and PTHrP, although many actions appear to be independent of adenylyl cyclase. Here we show that the PTH1R binds to Na(+)/H(+) exchanger regulatory factors (NHERF) 1 and 2 through a PDZ-domain interaction in vitro and in PTH target cells. NHERF2 simultaneously binds phospholipase C beta 1 and an atypical, carboxyl-terminal PDZ consensus motif, ETVM, of the PTH1R through PDZ1 and PDZ2, respectively. PTH treatment of cells that express the NHERF2 PTH1R complex markedly activates phospholipase C beta and inhibits adenylyl cyclase through stimulation of inhibitory G proteins (G(i/o) proteins). NHERF-mediated assembly of PTH1R and phospholipase C beta is a unique mechanism to regulate PTH signalling in cells and membranes of polarized cells that express NHERF, which may account for many tissue- and cell-specific actions of PTH/PTHrP and may also be relevant to signalling by many G-protein-coupled receptors.     

Carbon-dioxide-induced exocytotic insertion of H+ pumps in turtle-bladder luminal membrane: role of cell pH and calcium

The contents of endocytic vesicles and other intracellular organelles (such as Golgi and microsomes) are acidified by an electrogenic proton-translocating ATPase that is remarkably similar to that found in urinary epithelia. We recently found that the number of H+ ATPases in the apical plasma membrane of these epithelia is regulated by exocytotic insertion of endocytic vesicles whose membranes contain this H+ pump. Carbon dioxide, a major stimulus for urinary acidification, causes rapid fusion of these vesicles with the luminal membrane, thereby inserting these pumps there and increasing the rate of net transepithelial H+ secretion; CO2 also inhibits endocytic retrieval of the pumps from the luminal membrane. Such reciprocal regulation of endocytosis and exocytosis by a physiological modulator makes this system particularly attractive for studying the cellular events regulating membrane fusion. Here we present evidence that CO2 induces exocytosis by a cascade of events, the first step of which is cytoplasmic acidification. Cell acidification then increases calcium activity, which causes the fusion event.     

Voltage-dependent electrogenic chloride/proton exchange by endosomal CLC proteins

Eukaryotic members of the CLC gene family function as plasma membrane chloride channels, or may provide neutralizing anion currents for V-type H(+)-ATPases that acidify compartments of the endosomal/lysosomal pathway. Loss-of-function mutations in the endosomal protein ClC-5 impair renal endocytosis and lead to kidney stones, whereas loss of function of the endosomal/lysosomal protein ClC-7 entails osteopetrosis and lysosomal storage disease. Vesicular CLCs have been thought to be Cl- channels, in particular because ClC-4 and ClC-5 mediate plasma membrane Cl- currents upon heterologous expression. Here we show that these two mainly endosomal CLC proteins instead function as electrogenic Cl-/H+ exchangers (also called antiporters), resembling the transport activity of the bacterial protein ClC-e1, the crystal structure of which has already been determined. Neutralization of a critical glutamate residue not only abolished the steep voltage-dependence of transport, but also eliminated the coupling of anion flux to proton counter-transport. ClC-4 and ClC-5 may still compensate the charge accumulation by endosomal proton pumps, but are expected to couple directly vesicular pH gradients to Cl- gradients.     

Early Renal Morphological Changes in High-Cholesterol Diet Rats Model

In rats fed with high-cholesterol （HC） chow, the renal specimens were investigated by microscopy and enzymehistochemistry. The levels of serum lipids, 24 h urinary protein excretion （ UPE ）, N-acetyl-β-D-glucosaminidase （NAG） and Nitric Oxide （NO） were evaluated. Histological examination showed cell swelling, break-down and massive lipid deposition in renal tubules; perivascular and interstitial cell infiltration and mesangial cell proliferation. Enzymehistochemistry demonstrated that lactate dehydrogenase （LDH） activity in proximal tubular epithelial cells increased but succino dehydrogenase （SDH） activity decreased. The NO level in serum, urine and renal cortex were all decreased （p〈0.01）. Urinary NO, was negatively correlated with urinary NAG and UPE （r is -0. 525, 0. 529 respectively, p〈0.01）. This study shows that a HC diet can induce the early morphological changes in the whole kidney, particularly in the renal tubules. The decrease of NO is associated with the pathogenesis of hypercholesterolemia-induced renal injury.     

中华蟾蜍泌尿系统组织学的初步研究

1999年12月至2001年6月采用常规组织学方法对中华蟾蜍（Bufo gargarizans)的泌尿系统作了研究。结果表明：肾实质主要由肾小体和肾小管组成。肾脏中，后段的肾小体较多，多分布在肾脏周缘。肾小管由颈段，第一近曲小管，第二近曲小管，间段，第一远曲小管，第二远曲小管和集合管组成；输尿管位于肾脏后段外缘，其上皮为假复层柱状上皮或移行上皮；中华蟾蜍的膀胱为泄殖腔膀胱，膀胱内壁具大量发达的绒毛，绒毛表面是移行上皮，在其中分布有许多杯状细胞，膀胱具有重吸收水分的功能。     

甲状旁腺素(PTH)对氯化钾引起大白鼠输精管收缩的舒张作用

本研究探讨甲状旁腺素(PTH)对输精管平滑肌的舒张作用及其机制。不同剂量的氯化钾(20、40、60mM)引起大白鼠输精管的收缩,同样可被牛型—PTH(1—34)或人型—PTH(1—34)所舒张,而且均呈现剂量的依从关系。异丙肾上腺素舒张输精管的作用可被心得安所阻滞,但心得安或酚妥拉明均不抑制甲状旁腺素舒张输精管的作用,提示甲状旁腺素和异丙肾上腺素对输精管的舒张作用是通过不同的受体实现的。不同剂量的氯化钙引起输精管不同张力强度的收缩,甲状旁腺素对这种外源性钙引起的收缩呈现明显的抑制作用,说明甲状旁腺素有减少钙离子进入平滑肌组织的作用。     

肾草酸钙结石大鼠肾组织中Claudin-14的表达变化及意义

为构建大鼠肾草酸钙结石模型,从基因和蛋白水平初步探讨紧密连接蛋白-14(Claudin-14)在肾草酸钙结石大鼠肾组织中的表达变化及意义。采用30只雄性S-D大鼠随机分为对照组(n=15)和结石组(n=15),采用乙二醇和氯化铵灌胃诱导制作大鼠肾草酸钙结石模型。分别采用RT-PCR检测Claudin-14 mRNA表达;Western-Blotting检测Claudin-14蛋白表达;全自动生化分析仪检测各组大鼠24 h尿钙排泄。结果显示:光镜下结石组有大量结石结晶形成,且24h尿钙较对照组显著升高(P0.01),Claudin-14 mRNA表达较对照组显著升高(P0.01);Claudin-14蛋白在结石组高表达,在对照组未见表达;相关性分析发现结石组Claudin-14蛋白表达与24 h尿钙排泄呈显著正相关。由此可知,Claudin-14在肾草酸钙结石大鼠肾组织中高表达,可能是肾小管内尿钙增高,结石形成的机制之一。     

体外冲击波治疗复杂性上尿路结石

本文报告采用 ESWL 治疗复杂性上尿路结石57例的结果,全部碎石成功,无严重并发症发生。笔者在讨论中对采用体外冲击波碎石术治疗复杂性上尿路结石进行评价,并着重讨论其碎石治疗的原则以及输尿管石街的处理。     

Plasticity of functional epithelial polarity

The fundamental characteristics that allow vectorial transport across an epithelial cell are the differential sorting and insertion of transport proteins either in the apical or the basolateral plasma membrane, and the preferential association of endocytosis and exocytosis with one or the other pole of the cell. Asymmetrical cellular structure and function, being manifestations of terminal differentiation, might be expected to be predetermined and invariant. Here we show that the polarity of transepithelial H+ transport, endocytosis and exocytosis in kidney can be reversed by environmental stimuli. The HCO3- secreting cell in the cortical collecting tubule is found to be an intercalated cell possessing a Cl-/HCO3- exchanger in the apical membrane and proton pumps in endocytic vesicles that fuse with the basolateral membrane; the H+-secreting cell in the medullary collecting tubule has these transport functions on the opposite membranes. Further, the HCO3- -secreting cell can be induced to change its functional polarity to that of the H+-secreting cell by acid-loading the animal.