Objective To study the renal pathology and expression of vascular endothelial growth factor (VEGF) in rats with hypothyroidism caused by iodine deficiency.Methods The rats were randomly grouped into test group and normal group,and the test group was further randomly divided into model group and L-T4 group.Observation were made when the hypothyroid model was formed after maneuvering for 21 days and the medication had been taken for 56 days.The observed data included the morphogy of the kidney,serum TT3,TT4,TSH,blood urea nitrogen (BUN),serum creatinine (SCr),and VEGF expression.Results (1) After medication for 56 days,compared to normal group,BUN and SCr levels in L-T4 and model groups showed apparently increasing [BUN (5.55 ± 0.50,5.80 ± 0.66 vs 5.00 ± 0.24) mmol/L,P＜0.05 ; SCr (26.83 ± 0.75,27.0 ± 3.41 vs 22.5 ± 2.07) μmol/L,P＜0.05],and L-T4 group showed slightly lowered BUN and SCr levels compared with model group (P＞0.05).(2) By 21 days,in the model group there was moderate mesangial hyperplasia involving focal glomeruli.By 56th day of medication,entire glomeruli showed moderate mesangial hyperplasia with lessened number of dilated capillaries in the model group.However,in the L-T4 treated group,mesangial hyperplasia was much less and capillaries though dilated appeared wholesome,as compared with the model group.(3) By 21th day,the IODA value of VEGF in the model group was higher than that in the normal control (44.64 ± 14.99 vs 29.05 ± 9.14,P＜0.01).After medication for 56 days,there was no significant difference in the IODA value of three groups.Conclusions The characteristics of renal pathology in rats with hypothyroidism caused by iodine deficiency consist of mesangial hyperplasia and loss of glomerular capillaries.The expression of VEGF is raised early in the course of renal damage,but lowered thereafter.L-T4 can not reverse the sustained kidney damage and impaired renal function.

Objective To study the effects of integrated liposomal prostaglandin E_1(PGE_1)and calcium dobesilate on aristolochic acid nephropathy(AAN).Methods Forty-six patients of AAN who came from the First Affiliated Hospital,China Medical University from 2003 to 2005 were randomly divided into control group(PGE_1)and treatment group(PGE_1+calcium dobesilate).Scr and Hb were compared between two groups before and after the treatment.Results Scr had been decreased significantly and Hb had been increased in both groups,but more significantly in treatment group than in control group.Conclusion Integrated liposomal prostaglandin E_1 and calcium dobesilate can ameliorate renal function in AAN.

Objective To analyze the protein expression profile of human glomerular mesangial cells (HMCs) under high glucose and to characterize molecular functions and biological processes. Methods HMCs were divided into high glucose cultured group (30 mmol/L) and normal glucose cultured group (5 mmol/L). The total proteins were extracted after culture for 48 hours. The total proteins of the two groups were separated using two-dimensional fluorescence difference in gel electrophoresis (2-D DIGE) and analyzed using DeCyder 2-D difference analysis software. The differentially expressed proteins were further identified using in-gel digestion with trypsin, of which peptide extracts were prepared for MALDI-TOF-MS analysis. Protein identifications were searched in the NCBI protein database using the Mascot search engine. Results One hundred and forty-seven protein spots whose expression levels were significantly increased or decreased more than 1.5 folds under high glucose were identified. Ninety-six differentially expression protein spots were analyzed by peptide mass fingerprinting and 37 kinds of proteins were identified. The protein spots of phosphatidylethanolamine binding protein 1 (PEBP-1), granulysin,ATP synthase H + transporting mitochondrial FO complex subunit F2 were observed only in high glucose group. The expression of 24 proteins was up-regulated by high glucose, including eosinophil cationic protein, RGS membrane-interacting proteins 16 (MIR16), peptidyl-prolyl cis-trans isomerase, disks large homolog DLG2, breast cancer 2, early onset (BRCA2), Catechol-O-methyltransferase etc. The expression of 5 proteins was down-regulated by high glucose, including O-GlcNAc transferase-interacting protein 106 000 isoform 1, proteasome beta 6 subunit precursor,NEFA-interacting nuclear protein NIP30 etc. Conclusions Expression of 147 proteins in HMCs alters under high glucose. These proteins are involved in the regulation of cytoskeleton, glucose metabolism, cell division, gene transcription, signal transduction, phosphorylation, cell proliferation,apoptosis etc. In-depth analysis of these differentially expressed proteins' function and crosstalk is expected to provide an important experimental basis for clarifying the pathogenesis of diabetic nephropathy.

ObjectiveWith a GcneChip(R) cxpression analysis,98 known gencs and 31 exprcssed sequence tags (ESTs) were found to be differentially expressed between KK/Ta and BALB/c kidneys.To further screen the susceptibility genes for diabetic nephropathy,the temporal and spatial expression patterns of differentially expressed gene-adipsin were investigated.MethodsThe body weight,blood glucose,urinary albumin/creatinine ratio,and renal pathological changes of KK/Ta and BALB/c mice were measured at the 7,20,28 and 36 weeks of age.Total RNA was extracted from the kidney,heart,liver,lung,and brain.The temporal and spatial expression patterns of adipsin in diabetic KK/Ta mice were examined by competitive RT-PCR.The correlation analysis between adipsin expression and albuminuria level was carried out.ResultsThe mRNA expression of adipsin was found in the kidney,heart,lung,and brain,but not in liver.The expression of adipsin in diabetic KK/Ta mice at 20 weeks of age was significantly down-regulated in kidney,heart,and lung than that in age-matched BALB/c mice,and unaltered in brain.Adipsin expression in KK/Ta kidneys was significantly down-regulated with aging and negatively correlated to urinary albumin/creatinine ratio( r =-0.807,P ＜ 0.05).ConclusionsThe expression of adipsin mRNA was downregulated in kidney,heart,and lung in diabetic state.Adipsin expression in KK/Ta kidneys was negatively correlated to urinary albumin/creatinine ratio.It might be a candidate gene for diabetic nephropathy.

Objective The effects of candesartan,an angiotensin Ⅱ type 1 receptor blocker (ARB) were investigated on advanced glycation end-products accumulation and the receptor for AGE (RAGE) expression in type 2 diabetic KK/Ta mouse kidneys.MethodsKK/Ta mice(n=72)were random divided into three groups(n=24) and it was treated with candesartan [4 mg/(kg·d)] or vehicle from 6 or 12 to 28 weeks of age.BALB/c mice(n=24) treated with vehicle were used as controls.Body weight,blood pressure,blood glucose,urinary microalbumin,urinary creatinine and serum creatinine were measured every four weeks.At 28 weeks,renal expressions of carboxymethyllysine and RAGE were evaluated by immunohistochemistry and/or competitive RT-PCR.Results KK/Ta mice developed high body weight,high blood glucose,and high urinary microalbumin/creatinine ratio in KK/Ta mice at 28 weeks of age,and it was significantly higher than that of BALB/c mice [(427.49±89.37)mg/g vs (9.54±3.25)mg/g,P＜0.01 ].Protein and mRNA expressions of RAGE were upregulated in KK/Ta kidneys with increased immunostaining intensities of carboxymethyllysine.Candesartan treatment has markedly reduced urinary microalbumin/creatinine ratio [Early treatment group (32.18±9.41)mg/g,Late treatment group (53.20±7.26)mg/g,P＜0.01 ].Treatment with candesartan down-regulated the protein and mRNA expressions of RAGE and reduced the accumulation of carboxymethyllysine.There were no significant differences between the two treatment groups (from 6 or 12 weeks).ConclusionsThe results suggest that candesartan,an ARB,reduces advanced glycation end-products accumulation and subsequent albuminuria by down-regulating RAGE expression in type 2 diabetic KK/Ta mouse kidneys.

Objective To investigate the effects of angiotensin receptor blocker (ARB)losartan on the glomerular protein expression profile of spontaneous type 2 diabetic KKAy mice by two-dimensional differential gel eleetrophoresis and MALDI-TOF mass spectrometry.Methods 8-week-old spontaneous type 2 diabetic KKAy mice were randomly divided into losartan (10 mg·kg-1·d-1 given in drinking water) treatment group and non-treatment group.Eight-week-old C57BL/6 mice were used as normal control.The glomeruli were separated by magnetic bead perfusion through thoracic aorta at age of 20 weeks,then glomerular protein was extracted.The glomerular protein expression profile was investigated by CyDyes minimal fluorescence labelling,two-dimensional differential gel electrophoresis and MALDI-TOF mass spectrometry.Results KKAy mice developed higher body weight and blood glucose,higher urinary microalbumin creatinine ratio at age of 20 weeks than C57BL/6 mice at the same age (all P＜0.05).Losartan treatment markedly reduced urinary microalbumin creatinine ratio [(539.71 ±100.23)mg/g vs (728±177.19) mg/g],attenuated mesangial expansion and the thickening of glomerular basement membrane,but had no effect on the blood glucose.By DeCyder 2-D differential analysis software,62 protein spots of differential expression were found in glomeruli between losartan treatment and non-treatment KKAy mice at age of 20 weeks.Among them,41 proteins were identified by peptide mass fingerprinting.The expressions of 28 proteins were up-regulated by losartan treatment,including glycerokinase,sulfite oxidase,glycine amidinotransferase,adenosylhomocysteinase,etc.The expressions of 13proteins were down-regulaled by losartan treatment,including 3-mercaptopyruvate sulfurtransferase,ATP synthase subunit d,60 000 heat shock protein,stress-70 protein (alternative name 75 000glucose-regulated protein,GRP75),etc.Six differcntially expressed proteins were found in glomeruli between non-treatment KKAy mice and C57BL/6 mice,and the differential expressions were suppressed by losartan treatment,including dihydrolipoyllysine-residue acetyltransferase component of pyruvate dehydrogenase complex,succinyl-CoA ligase (GDP-forming) subunit beta,mitochondrial,ATP synthase subunit d,GRP75,nucleoside diphosphate-linked moiety X motif 19 and seleniumbinding protein 1.Conclusions Losartan significantly reduces the urinary protein excretion rate and renal pathological lesion of spontaneous type 2 diabetic KKAy mice,and suppresses the differential expression of mitochondrial ATP synthase subunit d,GRP75,selenium-binding protein 1,etc in glomeruli.Losartan may play a renoproteetive role by reducing glomerular mitochondrial reactive oxygen species genesis and inhibiting oxidative stress.

Objective To analyze the risk factors and correlation between clinical indicators and the four main pathological lesions of IgA ne?phropathy in the Oxford classification:mesangial hypercellularity(M0/1),endocapillary proliferation(E0/1),segmental sclerosis or adhesion(S0/1), and tubular atrophy/interstitial fibrosis(T0/1/2). Methods Clinical and pathological data were collected from 514 patients with biopsy?proven IgA nephropathy admitted in our hospital from February 17,2006 to October 11,2011. These patients were all above 18 years old. Cases with sec?ondary causes of mesangial IgA deposition were excluded,such as Henoch?chonlein purpura,ankylosing spondylitis and psoriasis et al. The inde?pendent risk factors affecting the pathological classification were analyzed by Spearman rank correlation analysis and two?category and multi?classi?fication logistic regression using SPSS 17.0 statistical software. Results In 514 IgAN patients,the ratio of males to females was 1.06:1. The aver?age age was 35.70±11.99 years,and the average disease duration was 18.31±30.42 months. M0E0S0T0 was the major pathologic classification of isolated hematuria. Chronic kidney disease(CKD)stage,24 hours proteinuria,albuminuria,urine transferrin and IgG levels were positively corre? lated with M lesion;serum albumin,C3 and PLT showed a negative correlation with M lesion. Twenty four hours proteinuria and blood platelet count were the independent risk factors for M lesion. As shown by stratified analysis ,the proportion of M1 in cases with 24 hours proteinuria≥3.5 g/d is much higher than that in cases with non?nephrotic range proteinuria. Age,systolic blood pressure,uRBC,24 hours proteinuria,albuminuria urine transferrin and IgG levels were positively correlated with E lesion,Duration,serum albumin showed a negative correlation with E lesion. Age and duration of nephritis were independent risk factors for E lesion. 73.3%of patients that above 60 years old showed endothelial proliferation. CKD stage,24 hours proteinuria were positively correlated with S lesion. Age,CKD stage,systolic blood pressure,diastolic blood pressure,C4,TC, LDL?C,CRP,Fib,UA,Cys?C and 24 hours proteinuria,urineβ2?microglobulin,albumin,transferrin and IgG levels were positively associated with T lesion;hemoglobin,serum albumin,serum IgG showed a negative correlation with T lesion. Infection history,high CRP levels,DBP more than 90 mmHg,hypoalbuminemia,high low density lipoproteinemia,and anemia were independent risk factors for T lesion. Conclusion Twenty four hours proteinuria,blood platelet count,age,duration of nephritis,hypoalbuminemia,anemia,hyperlipidemia,DBP≥90 mmHg and high CRP lev?els were risk factors for the Oxford classification of IgA nephropathy. Renal biopsy should be carried out in time to make clear the pathological clas?sification and individual treatment,so as to improve the prognosis.

Objective To observe the epithelial mesenchymal transition (EMT) of podocyte induced by high glucose,and to explore the potential protective mechanism of ursolic acid (UA).Methods The podocytes cultured in vitro were divided into four groups:normal group (glucose 5.5 mmol/L),mannitol group (glucose 5.5 mmol/L+mannitol 19.5 mmol/L),high glucose group (glucose 25 mmol/L) and UA group (glucose 25 mmol/L + UA 5 μmol/L).Podocyte morphology changes were observed by inverted phase contract microscope.The expression of zonula occludens-1 (ZO-1) and α-smooth muscle actin (α-SMA) were detected by immunofluorescence.The expressions of β-catenin and glycogen synthesis kinase-3β (GSK3β) were detected by Western blotting.The expressions of Wnt1,Wnt3a,Wnt5a,Wnt5b and GSK3β were detected by real-time PCR.Results Podocytes showed irregular arborization shape in normal glucose and transited to longer cobblestone-like shape as mesenchyme cell by high glucose culture.Compared with normal group,the expression of ZO-1 protein was down-regulated and the expression of α-SMA was up-regulated by high glucose culture (P ＜ 0.05).The expression of Wnt5a mRNA was down-regulated;β-catenin mRNA and protein were up-regulated (P ＜ 0.05);and GSK3β protein was down-regulated by high glucose culture (P ＜ 0.05).Compared with high glucose group,ursolic acid inhibited podocyte EMT,up-regulated the expression of ZO-1 protein,Wnt5a mRNA,GSK3β (P ＜ 0.05),and down-regulated the expressions of α-SMA protein,β-catenin mRNA and protein (P ＜ 0.05).Conclusion Ursolic acid attenuates high glucose induced epithelial mesenchymal transition of podocyte by inhibiting Wnt/β-catenin signaling pathway.

Objective To investigate the expression of Notch signaling molecules, transforming growth factor-β (TGF-β) and fibronectin (FN) in mesangial cell induced by high glucose, and the underlying mechanism of cordyceps sinensis.Methods Rat glomerular mesangial cells were divided into following groups: normal control group (5.5 mmol/L glucose), hypertonic control group (5.5 mmol/L glucose+ 19.5 mmol/L mannitol), high glucose group (25.0 mmol/L glucose), DAPT inhibitor group (25.0 mmol/L glucose + 1 μmol/L DAPT), cordyceps sinensis intervention group (25.0 mmol/L glucose+10 mg/L cordyceps sinensis).Cell proliferation was detected by MTT.The protein and mRNA expression of Notch signaling molecules (Notch1, Jagged1 and Hes1), TGF-β and FN was detected by Western blotting and real time PCR.Results Compared with normal control group, high glucose induced mesangial cell proliferation, as well as the mRNA and protein expression of Notch1, Jagged1, Hes1, TGF-β1 and FN was up-regulated in high glucose group (all P ＜ 0.05).Compared with that in high-glucose group, DAPT and cordyceps sinensis inhibited high glucose-induced mesangial cell proliferation and down-regulated the mRNA and protein expression of Notch pathway, TGF-β1 and FN (all P ＜ 0.05).Conclusion By inhibiting the abnormal activation of Notch signaling pathway and TGF-β signaling pathway, cordyceps sinensis may alleviate high glucose-induced mesangial cell proliferation and reduce extracellular matrix accumulation, thus protecting kidney.