用傅立叶红外光谱测定泌尿系结石中的草酸钙

对泌尿系结石的组成进行结构分析,为分析结石病的产生原因及预防复发提供参考,在临床诊断和治疗上具有重要意义.傅立叶红外光谱是研究泌尿系结石构造的一种较理想的分析手段.用红外光谱仪定量分析泌尿结石中草酸钙采用的内标法、吸光度比值法和一阶导数法进行了比较,建立的分析方法可以简便对草酸钙混合结石中一水草酸钙(COM)和二水草酸钙(COD)进行定量测定,分析结果可初步满足临床诊断的要求.     

Mimicking the growth of a pathologic biomineral: shape development and structures of calcium oxalate dihydrate in the presence of polyacrylic acid

The morphogenesis of calcium oxalate hydrates in aqueous solutions was investigated by varying the pH, oxalate concentration, and the concentration of the sodium salt of polyacrylate (PAA). With increasing amounts of PAA in solution, the shape of tetragonal calcium oxalate dihydrate (COD) changes from bipyramidal through elongated bipyramidal prisms to dumbbells and finally reverts to rodlike tetragonal bipyramidal prisms. PAA is incorporated into the prismatic zones of the growing COD crystals, thereby reducing the growth rate of the {100} faces along the <100> direction. Dumbbells start to develop through "non-crystallographic" branching from the prism faces and the formation of "multiple head" crystals. Adsorption of PAA on the rough surfaces of the splitting individuals supports the selection of new subindividuals and leads to the formation of core-shell patterns. The various shapes and structures of the biomimetic COD/PAA crystals and aggregates are closely related to the well-known "pathologic" individuals observed in the urine of patients with urinary disease (including urinary stones).     

二水草酸钙的形成及其对尿路结石的防治

二水草酸钙(COD)是泌尿系结石的主要成分之一,其成核、生长和聚集过程与尿石的形成密切相关。本文结合我们近年来的工作,综述了生物膜及其模拟膜对COD的调控作用,尿大分子、焦磷酸盐、多磷酸盐、柠檬酸、酒石酸及表面活性剂等尿小分子的诱导作用,过饱和度、化学计量条件、pH值、离子强度、温度等体系参数对COD形成的影响;讨论了COD的形貌、晶面电荷及其与细胞膜粘附的研究进展。从临床上预防和治疗草酸钙结石的角度,综述了有利于COD形成的因素。     

Thermal, UV, FTIR, and XRD studies of urinary stones

Various crystals are seen in human urine. Oxalate, Phosphate, Uric acid, and Urate crystals are generally seen in urinary calculi. Calcium stones are most common, comprising 75 % of all urinary calculi. They may be pure calcium oxalate or calcium phosphate or a mixture of both. Many stones are not homogeneous. Low calcium intake increases the intestinal absorption of calcium, thus decreasing the amount of calcium available in the intestinal tract to form insoluble complexes with Oxalate. Consequently, a higher amount of oxalate is available for intestinal absorption and as a result, urinary oxalate excretion increases. Mineral water consumption did not reduce urinary oxalate excretion. High urinary excretion and concentration of magnesium decrease both the nucleation and growth rates of calcium oxalate crystals in urine, because of the higher solubility of magnesium oxalate compared with calcium oxalate. Analytical results show calcium oxalate to be one of the major inorganic components of renal stones and found to be present in almost all kidney and bladder stones. About 39.5 % of the total composition of the calculi is found to contain purely calcium oxalate and also hydroxyl apatite. The ten samples are a mixture of calcium oxalate and phosphate stones. Four samples are calcium oxalate as major composition and the remaining are calcium phosphate as major composition. These kidney stones are taken photographically and size of the stone are measured using optical microscopy. These qualitative analyses are also confirmed by UV, FTIR, DSC, and XRD analysis.     

不同介质凝胶体系中草酸钙结晶的研究

研究了五种不同介质（水、氯化钠、合成尿、正常人尿液和尿石患者尿液）的凝胶体系中草酸钙（CaC2O4）晶体的生长,及各种体系中防石药物柠檬酸钾（K3cit）对CaC2O4生长的影响.没有加K3cit时,CaC2O4晶体以一水草酸钙（calcium oxalate monohydrate, COM）为主要物相,但在氯化钠和合成尿的凝胶体系中同时出现了二水草酸钙（calcium oxalate dihydrate, COD）和三水草酸钙（calcium oxalate trihydrate, COT）,肾结石患者尿液中出现COD,而正常人尿液中没有COD和COT生成.加入K3cit后,水、氯化钠和合成尿介质的凝胶体系中,COT的含量显著增加,患者尿液中产生大量COD,而正常人尿液中出现了少量的COD和极个别的COT.COT的增加与低温、体系中高的离子强度及金属离子等因素有关.K3cit具有诱导COD和COT的生成、减小COM晶体比表面积的作用,从而有利于防治草酸钙结石的形成.     

Inhibition of the Crystal Growth and Aggregation of Calcium Oxalate by Algae Sulfated Polysaccharide In-vitro

Kidney stone disease causes substantial suffering and occasional renal failure. The content of calcium oxalate (CaOxa) is up to 70-80% in the stones1,2. However, the mechanism of the formation of urinary stones is not yet clearly understood and the questi…     

脂质体中不同种类羧酸钾对草酸钙晶体生长的调控作用

首次研究了狼磷脂-水脂质体中不同种类羧酯钾对草酸钙晶体生长的调控作用 。加入一元DAc只诱导一水草酸钙（COM）生成。二元K_2tart在其浓度大于1. 0mmol/L时可以诱导三水草酸钙（COT）生成。而加入三元的K_3cit和四元的 K_2edta后， 在不同的浓度下，可以分别诱导COM，二水草酸钙（COD）和COT的生 成。在低浓度（—3.3-17mmol/L）范围，， COD含量达到100%；而在较高浓度(＞ 17mmol/L)时，COD减少，COT含量增加。在不同的浓度区间，无论是COM含量减少， 还是COT含量增加，或者是COD含量的先增加后减少，均与该羧酸钾浓度的对数呈线 性关系。不同羧酯钾抑制COM生长并诱导COD形成的能力顺序为：K_3cit＞K_2edta ＞＞K_2tart-KAC,诱导COT生长的能力顺序为：K_2tart＞＞K_3cit＞K_2edta＞＞ KAc.由此推测抑制草酸钙结石形成的潜在效率依次为：K_3cit＞K_2edta＞＞ K_2tart＞＞KAc.     

凝胶体系中不同结构羧酸盐对草酸钙生物矿化的影响

采用双扩散法研究了凝胶体系中四元羧酸盐(Na2EDTA)、三元羧酸盐柠檬酸钠（Na3cit）、二元羧酸盐酒石酸钠（Na2tart）和一元羧酸盐醋酸钠（NaAc）对草酸钙（CaOx）结晶的影响.抑制一水草酸钙（COM）聚集的能力为:Na2EDTA >Na3cit >Na2tart >NaAc；诱导二水草酸钙（COD）的能力为：Na3cit >Na2tart >Na2EDTA >NaAc.羧酸的抗衡阳离子影响CaOx的结晶. H3cit、Na3cit和K3cit抑制COM聚集和诱导COD形成的能力均为:K3cit-Na3cit >H3cit.无论是诱导COD生成，还是抑制COM聚集，均可以减小结石形成的几率，对临床上防治结石具有积极的意义.     

土家族药物天葵化石汤提取液抑制草酸钙结石形成的化学模拟

采用扫描电子显微镜(SEM)和X射线衍射法(XRD)研究了土家族药物天葵化石汤(Tiankui)的提取液对尿石晶体草酸钙(CaOxa)成核、生长的抑制作用及其对二水草酸钙(COD)的稳定作用. 当Tiankui浓度小于30 mg/mL时, 主要生成一水草酸钙(COM), 但可使COM晶体棱角圆钝. 随着Tiankui浓度增加至45和75 mg/mL, 可分别诱导95%和100%的COD晶体生成. Tiankui还可以稳定COD在水溶液中的存在, 没有Tiankui存在时, COD在48 h内全部转化为COM晶体, 而在12 mg/mL Tiankui存在下, COD仅有20%转化. 从Tiankui提取液的活性组分、与草酸钙不同晶相间的吸附差异、化学配位等角度讨论了其抑制CaOxa生长和诱导稳定COD的化学基础.     

海藻异枝麒麟菜多糖抑制尿石盐草酸钙晶体生长的研究

采用扫描电子显微镜(SEM)、 红外光谱(FTIR)和X射线衍射(XRD)方法研究了从海藻异枝麒麟菜中提取的硫酸多糖(ESPS)对草酸钙(CaOxa)晶体生长的影响. 结果表明, 添加ESPS能抑制一水草酸钙(COM)的生长, 同时诱导二水草酸钙(COD)晶体的形成. 随着ESPS的质量浓度从0增加到0.03和0.50 mg/mL, COD的质量分数从0分别增加到10%和55%; COM的(1 01)晶面加强, (020)晶面减弱直至消失, 并从三维晶体转变为棱角圆顿的四角形片状晶体; COM和COD的尺寸均明显变小. 这些结果表明, ESPS是抑制CaOxa结石形成的一种潜在药物.     

大豆多糖对尿石矿物草酸钙形成的调控作用

采用水提法从大豆中提取了大豆多糖(SPS),其多糖百分含量为97.38%;平均相对分子质量为115,200,羧基百分含量为13.8%。采用X射线衍射法(XRD)、红外光谱(FTIR)、扫描电子显微镜(SEM)和电感耦合等离子体发射光谱仪(ICP)研究了水溶性大豆多糖(SPS)对草酸钙晶体生长的调控作用。SPS大豆多糖不但可以抑制一水草酸钙(COM)晶体的聚集,而且可以诱导二水草酸钙(COD)晶体形成。当SPS浓度由0增大到0.50g·L-1时,草酸钙晶体由COM完全转变为COD晶体;并且随着多糖浓度的增大,溶解在溶液中的钙离子(可溶性钙)浓度显著增加,草酸钙沉淀的物质的量逐渐减少。这些结果表明,SPS有望成为预防草酸钙结石的一种绿色药物。     

硅凝胶体系中不同结构羧酸钾对草酸钙结晶的影响

采用扫描电子显微镜(SEM)和X射线衍射(XRD)方法研究了硅凝胶体系中不同种类羧酸钾对草酸钙(CaC₂O₄)晶体生长的调控作用. 加入一元醋酸钾(KOAc)只生成一水草酸钙(COM)晶体; 三元柠檬酸钾(K₃Cit)和四元乙二胺四乙酸二钾(K₂EDTA)可诱导二水草酸钙(COD)形成, 且随着其浓度增加, 对COD的诱导能力增加, 而二元酒石酸钾(K₂Tart)同时诱导了COM, COD和三水草酸钙(COT)生成. 随着结晶温度降低, 多元酸钾可以进一步减小COM晶体的比表面积, 增加COD的百分含量, 但K₂Tart诱导COT的能力减弱. 由于诱导COD和COT晶体形成、减小COM的比表面积均有利于防止草酸钙尿石的形成, 因此, 多元羧酸钾可用于草酸钙结石的预防和治疗.     

玉米须提取液对尿液中草酸钙晶体形成的影响

本文采用X射线衍射(XRD)、红外光谱(FTIR)、扫描电子显微镜(SEM)等方法分析了玉米须提取液对正常人尿液中草酸钙晶体形成的影响,通过电导率法研究了草酸钙晶体生长的动力学过程,以及从生物矿化的角度对玉米须提取液影响尿液中草酸钙晶体的可能机理进行了探讨。由于玉米须提取液中有机酸或多糖的羟基、羰基等通过配位作用与Ca²⁺结合形成可溶性配位化合物,减少了Ca²⁺与Oxa^2-的结合能力,从而抑制了CaOxa的成核和生长。同时,可能由于玉米须提取液中有效成分与二水草酸钙(COD)的吸附点键合,增强了COD晶体在溶液中的热力学稳定性,进而抑制了COD晶体向热力学更稳定态的一水草酸钙(COM)晶体转变。结果显示,这种抑制作用随玉米须浓度增大而增大,且COD晶体尺寸随着玉米须浓度的增大而减小。玉米须抑制COD晶体向COM晶体转变的作用为开发预防和治疗尿结石的药物提供了启示。     

沙苑子提取液对不同体系中草酸钙晶体生长影响的研究

通过与水、氯化钠、正常人尿液体系的比较,重点研究了结石患者尿液体系中加入中药沙苑子提取液对草酸钙晶体生长的的影响,利用SEM,FTIR和XRD等测试手段对所得晶体进行表征。结果发现:在结石患者尿液体系中形成的草酸钙晶体为一水草酸钙(COM)晶体,而在这4种体系中加入沙苑子提取液后,只形成二水草酸钙(COD)晶体,表明沙苑子提取液能抑制COM晶体生长,并且随着沙苑子提取液浓度增大,抑制作用增强。沙苑子抑制草酸钙晶体生长的可能机理进行了探讨。     

酒石酸镁对水合草酸钙晶体生长的调控作用

本文在凝胶模拟体系中研究了Mgtart对草酸钙生长、 聚集和晶型、 晶相等的影响, 以期阐明Mgtart防治泌尿系结石形成的可能机理, 为临床医学提供参考.     

Simulation of calcium oxalate stone in vitro

Urolithiasis constitutes a serious health problem that affects a significant section of mankind. Between 3% and 14% of the population, depending on the geographical region, suffer from this illness[1]. For example, the incidence of urolithiasis in Florida in the United States of America was 15.7 in 100000 people and increased to 20.8 in 1996. Urolithiasis remains a major medical prob-lem in China, especially in Guangdong Province. A survey in 1997 in Shenzhen City, the most southern city i…     

Analysis of calcium, oxalate, and citrate interaction in idiopathic calcium urolithiasis in children

The majority of urinary stones in children are composed of calcium oxalate. To investigate the interaction between urinary calcium, oxalate, and citrate as major risk factors for calcium stones formation, their 24-h urinary excretion was determined in 30 children with urolithiasis and 15 normal healthy children. The cutoff points between children with urolithiasis and healthy children, accuracy, sensitivity, and specificity for each risk factor alone as well as for all three taken together were determined. OneR and J4.8 classifiers as parts of the larger data mining software Weka, based on machine learning algorithms, were used for the determination of the cutoff points for differentiation of the children. The decision tree based on J4.8 classifier analysis of all three risk factors together proved to be the best for differentiating stone formers from normal children. In comparison to the accuracy of the differentiation after calcium and oxalate of 80% and 75.6%, respectively, the decision tree showed an accuracy of 97.8%. Even when its stability was tested by the leave-one-out cross-validation procedure, the accuracy remained at a very acceptable percentage of 93.2% correctly classified patients. J4.8 classifier analysis gave a look inside urinary calcium, oxalate, and citrate interaction. Urinary calcium excretion was shown as the most informative in discrimination of the children with urolithiasis from healthy children. However, it was shown that oxalate and citrate excretions might influence the stone formation in a subpopulation of the stone formers. In patients with low urinary calcium, a major role in lithogenesis belongs to oxalate, in some of them alone and in others in conjunction with citrate. Decreased urinary citrate excretion in the presence of increased oxalate excretion may lead to stone formation.     

不同晶相草酸钙超细微晶的合成及其与硫酸软骨素A的相互作用

采用配位沉淀法制备了二水草酸钙(COD)和一水草酸钙(COM)超细微晶,其尺寸分别为150nm和320nm。通过扫描电子显微镜(SEM)、X射线衍射(XRD)、红外光谱(FTIR)、纳米粒度仪(Nano-ZS)和紫外-可见吸收光谱(UV-Vis)表征了这两种草酸钙微晶。研究了尿大分子硫酸软骨素A(C4S)对上述COM和COD微晶ξ电位、粒径、聚集程度和紫外吸光度的影响。随着cC4S从0增加到1.0g·L-1,COD微晶的ξ电位从-9.7mV减小到-46.1mV,COM微晶的ξ电位从-15.9mV减小到-49.0mV;微晶表面ξ电位变负后,有利于稳定溶液中悬浮的微晶。在水溶液中,COD和COM微晶均存在显著的聚集现象,而C4S的存在可抑制COD微晶的聚集,并在浓度为0.05g·L-1时抑制效果最好。由于尿液中存在大量草酸钙微晶,本研究有助于阐明草酸钙结石的形成机理和C4S对草酸钙结石形成的抑制作用。     

肾上皮细胞损伤使草酸钙晶体黏附增强的分子机制

研究了非洲绿猴肾上皮细胞(Vero)在损伤前后与一水合草酸钙(COM)和二水合草酸钙(COD)晶体的黏附作用及其引起的细胞反应,探讨了肾结石形成机理.COM和COD晶体与损伤细胞的黏附加重了细胞的过氧化损伤程度,导致损伤细胞的活力进一步降低,乳酸脱氢酶(LDH)释放量和活性氧(ROS)进一步增加,坏死细胞数量进一步增多,细胞体积缩小,并出现凋亡小体.COM晶体对细胞的损伤能力显著大于COD晶体.扫描电子显微镜(SEM)观测结果表明,损伤组Vero与COM微晶的黏附作用显著强于对照组,且能促进COM微晶的聚集.共聚焦显微镜观测结果表明,Vero损伤后,其表面表达的晶体黏附分子透明质酸(HA)显著增加,HA分子是促进微晶黏附的重要原因.细胞表面草酸钙的黏附量和晶体聚集程度与细胞的损伤程度成正相关.本文结果从分子和细胞水平上提示,细胞损伤是导致草酸钙肾结石形成的重要因素.