ABSTRACT
Objective Tamm-Horsfall protein (THP) may play a role in kidney stone formation.The article aimed to conduct a preliminary study on the role of THP in kidney stone formation by investigating the changes of THP in rat urine, pathological changes of renal tissue and the formation of calcium salt crystals after establishing CNPs rat model of kidney stones.Methods Stone samples of 40 patients from February to June 2015 in our department were collected to establish the model of CNPs-induced kidney stone in rats and prepare CNPs suspension.48 SD rats were randomly divided into experimental group (group A) and blank control group (group B).Group A were injected with CNPs and the same amount of sterile saline injection in the group B.The urine of rats was collected after injection at 3h, 6h, 12h, 24h, 1w, 2w, 4w and 8w.ELISA were applied to detect THP levels in the urine.Then the rats were killed to take the kidney tissue.HE staining was used to investigate the pathological changes of the cells and evaluate the formation of the calcium salt crystals.Results THP levels in group A at 24h, 1w, 2w, 4w and 8w ([166.03±3.02], [173.50±1.78], [174.55±2.05], [176.54±2.45], [177.11±1.76]pg/mL) were significantly higher than that at 3h(165.89±2.23pg/mL)(P<0.05), which was the same case in comparison with those of group B ([157.65±2.22], [156.54±1.43], [159.45±3.21], [158.63±2.98], [157.33±2.05]pg/mL).Compared with the calcium salt crystal score at 6h (1 point), the scores at 3,6,12,24h (average score 2 points) increased.At 2w the score increased significantly to 3 points and reached the top score(6.7points) at 8w, which was of significant difference.The score of calcium salt crystals was in positive correlation with THP content (r=0.843,P<0.05).Conclusion THP in urine may contribute to the aggregation of calcium salt crystals and the formation of kidney stones.
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Objective To establish renal calculus rat model induced by calcifying nanoparticles .Methods We extracted and cultivated calcifying nanoparticles from clinically diagnosed patients with nephrolithiasis .Forty adult male SD rats were divided into 4 groups (n=10 each):group A(given an intravenous injection of calcifying nanoparticles) ,group B(given an intravenous injection of calcifying nanoparticles and taken Shenshitong granules) ,group C(given an intravenous injection of calcifying nanoparticles and intraperitoneal injection of gallium nitrate) ,group D (as normal control) .Eight weeks later ,all rats were sacrificed .The kidneys were examined for pathology .Results Histopathological studies revealed calcifying nanoparticles induced renal tubular epithelial cells fatty degeneration and renal tubular crystallization ,and caused renal interstitial infiltration with inflammatory cells .The forma‐tion of renal crystals in group A was more obvious than those in group B and C(P0 .05) .Conclusion Calcifying nanoparticles could cause renal tubular epithelial cells injury ,which leads to the formation of kidney stones .
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Objective To explore the relation between thyroid nodular and thyroid calcification .Methods The clinical data of 430 cases with nodular goiter admitted from January 2011 to December 2013 were reviewed.All cases were performed ultrasonography and pathologic examination .Results Of 430 cases,there was thyroid carcinoma in 49 cases(11.4%),including microcalcification in 30 cases(61.2%) and macrocalcification in 3 cases(6.1%);benign lesion in 381 cases(88.6%),including microcalcification in 24 cases(6.3%) and macrocalcification in 39 cases(10.2%).The difference was remarkable (P=0.000).There was remarkable difference in malignant calci-fied nodules ratio between the patients with microcalcification and macrocalcification (55.6%vs 7.1%,P=0.000), between the patients aged 45 years below and above(48.8%vs 22.6%,P=0.007),between the patients with solita-ry and multiple nodules(45.0%vs 16.7%,P=0.005).But there was no statistic difference in malignant calcified nodules ratio between the male and female groups (24.2% vs.39.7%,P=0.130).Conclusion Microcalcification is a highly specific sign to predict thyroid carcinoma in ultrasonography exam .Positive intervention will be performed immediately when calcification appears in thyroid nodules .
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Objective To observe the effects of different concertrations of calcifying nanoparticles (CNP) on growth and proliferation of human renal tubular epithelial cells (HK-2),and to explore the effects of autophagy of human renal tubular epithelial cells induced by CNP. MethodsNormal growth HK-2 cells were cultured in vitro and the culture medium was changed to serum-free medium after cell growth to confluence,CNP with different concentrations were then added and the cell proliferation inhibition was measured by MTT assay after 12,24,48 and 72 h co-incubation.EGFP-LC3 express plasmid was transfected into HK-2 cells,CNP with certain concentration was then added after 24h incubation.The formation of autophagosomes and their adherence to cells were observed by electron microscopy,and EGFP-LC3 staining was used to detect the formation of autophagosome by flourescence microscopy after 3,6,24 and 48 h co-incubation. ResultsCNP presented a concentration-dependent cell proliferation inhibition on HK-2 cells,the inhibitory ratio in cell proliferation was 14.5 ％ after 48 h co-incubation when CNP density was 0.05 mg/ml,and the inhibitory ratio was 21.5％ after 72 h co-incubation when CNP density was 0.1 mg/ml.Autophagosome formation and CNP adherence to cell surface could be observed by electron microscopy 6 h after CNP was added to the DMEM medium.The LC3-positive dots were observed in HK-2 cells incubated with CNP.Conclusions Autophagy of HK-2 could be activated by co-incubation with CNP,which may play an important role in the formation of nephrolithiasis.
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ObjectiveTo observe the interaction of the calcifying nanoparticles (CNP) with human renal tubular epithelial cells (HK-2) in vitro,to observe and investigate the mechanisms of HK-2 injury induced by CNP,and to explore the potential role of CNP in the formation of Calcium oxalate kidney stones.MethodsHuman renal tubular epithelial cells were cultured in vitro and CNP was then added to the culture medium,the cell-crystal reaction was detected by light microscopy and transmission electron microscopy (TEM).To investigate the oxidative stress,NADPH oxidase inhibitor apocynin was chosen as the intervener.The levels of LDH,MDA,HA in the mediums after 24 h were assessed. ResultsCNP could induce changes of the HK-2.Adhesion and phagocytosis of CNP by the HK-2 were observed under TEM.After 24 h,the levels of LDH,MDA,HA were significantly different among the 4 groups ( P ＜ 0.05 ). ConclusionsHK-2 has abilities of adhering and phagocyting with CNP.CNP can cause damage induced by oxidative stress of HK-2.