significantly increased (all P < 0.01). Following SREBP-1 was down-regulated by siRNA, high glucose-stimulated TGF-β1 and FN protein expressions were decreased by 17.9% and 24.6% ,respectively(all P<0.01).

AIM: To study the effect of high fat diet on the expression of sterol regulatory element biding protein-1 (SREBP-1) and transforming growth factor β_1 (TGF-β_1) in renal tubular cells and rosiglitazone intervention. METHODS: Wistar rats were treated with high fat diet and rosiglitazone for 3 months. The serum glucose, serum insulin and serum triglyceride were detected. Oil Red O staining was used to observe the renal lipid deposit and Masson staining was for the detection of ECM accumulation. SREBP-1, TGF-β_1 and FN protein were determined by the methods of immunohistochemistry and Western blotting. SREBP-1 mRNA was detected by in situ hybridization. RESULTS: Rosiglitazone prevented effectively the increase in serum glucose, serum insulin and serum triglyceride resulted from high fat diet. High fat diet led to lipid droplet formation in renal tubular cells and interstitial ECM accumulation, which was decreased by rosiglitazone treatment. Compared to normal rats, SREBP-1 protein and SREBP-1 mRNA showed high expressions in high fat diet rats that were lowered by rosiglitazone. The precursor segment and mature segment of SREBP-1 protein were decreased by 27.39% and 27.32%. Similarly, the high expressions of TGF-β_1 and FN protein in kidney of high fat diet rats were also prevented by rosiglitazone intervention. Compared to high fat diet rats, the expression of TGF-β_1 in rosiglitazone treatment rats was lowered by 19.14%. CONCLUSION: Rosiglitazone prevents effectively the over-expression of SREBP-1 and TGF-β_1 in renal tubular cells, and decreases lipid accumulation and ECM production in rats fed with high fat diet.

Aim To investigate the effects of valsartan on the expression of TGF-?1 mRNA and the activation of p38 mitogen-activated protein kinase(p38MAPK),mitogen activated protein kinase kinasse3/6(MKK3/6)and cAMP response element-binding protein1(CREB1) in glomerular mesangial cells under high concentration of glucose.Methods High concentration glucose and valsartan were used to stimulate the cultured rat GMCs in vitro.The protein expressions of p38 MAPK,CREB1,p-p38 MAPK,p-MKK3/6 and p-CREB1 were observed by Western blot analysis.TGF-?1 and fibronectin(FN) mRNA were measured by reverse transcription and polymerase chain reaction(RT-PCR).The protein synthesis of FN and type Ⅳ collagen in the supernatants of the GMCs was detected by radioimmunoassay.Results Compared with low glucose control group,the expressions of p-p38 MAPK,p-MKK3/6,p-CREB1 protein,TGF-?1 and FN mRNA,FN and type Ⅳ collagen in the supernatants were significantly increased in GMCs under high concentration glucose medium.The expression levels of p-p38 MAPK,p-MKK3/6 and p-CREB1 were significantly lower in the valsartan group than those in the high concentration glucose group.So did the mRNA of TGF-?1 and FN.The concentration of FN and type Ⅳ collagen in the supernatants in the valsartan group was lower than that in the high concentration glucose control group.Conclusion Valsartan can inhibit over production of TGF-?1 and ECM proteins in GMCs under high concentration of glucose,partly by regulating the phosphorylation of p38 MAPK,MKK3/6 and CREB1.

Objective To investigate the impact of limbal stem cell transplantation in the treatment of pterygium on the tear film function in patients with type 2 diabetes mellitus. Methods All 70 cases of primary pterygium patients with type 2 diabetes mellitus in Beijing Changping district Hospital from January 2012 to December 2014 were chosen. Basal tear secretion, tears river area and tear film break-up time of their two eyes were measured. Excision of ptery-gium combined with limbal stem cell transplantation was conducted. Basal tear secretion,tears river area and tear film break-up time before surgery, two weeks and two months after surgery were measured respectively, and the impact of limbal stem cell transplantation in the treatment of pterygium on the tear film function in patients with type 2 diabetes mellitus was analyzed. Results The differences of preoperative Schirmer test and tear river area of two eyes were not statistically significant(P>0.05),and the difference of preoperative tear film break-up time of two eyes was statistical-ly significant(P<0.05). The differences of the operative eyes Schirmer test and tears river area before and after surgery were not statistically significant(P>0.05),and the difference of preoperative and postoperative tear break-up time was statistically significant (P<0.05). Conclusion The tear film after the limbal stem cell transplantation in the treatment of pterygium is more stable than that before surgery, and the limbal stem cell transplantation can improve tear film function in patients with type 2 diabetes mellitus.

Aim To investigate the time-dependent effect of insulin on the expression of SREBP-1(sterol regulatory element binding protein-1),FAS(fat acid synthase)and lipid droplet formation in HKC cells(human renal proximal tubular epithelial cells line).MethodsHKC cells were respectively treated with 100 nmol·L~(-1) insulin for 0,2,4,6,12 h and 24 h.The analysis of SREBP-1 and FAS mRNA was performed by RT-PCR and the expression of SREBP-1 protein was detected by Western blot and immunocytochemistry.Furthermore,Oil Red O staining was used to determine cellular lipid droplet formation.ResultsCompared with HKC cells of 0 h group,there was no difference of SREBP-1 and FAS mRNA in HKC cells of 2 h group.However,the expression of SREBP-1 and FAS mRNA was significantly increased in HKC cells of 4,6 h and 12 h group.Further,the most expression of SREBP-1 and FAS mRNA was at 6 h group and was respectively increased by 3.578 and 4.272 times compared with 0 h group.The results of Western blot showed that the precursor and mature segments of SREBP-1 protein in 4,6 h and 12 h group HKC cells were increased and those of 6 h group HKC cells were the highest and about 4.106 and 5.167 times than those of 0 h group HKC cells.Immunocytochemistry presented the result that SREBP-1 protein was located in the plasma and the expression of 4,6 h and 12 h group HKC cells was significantly higher than that of 0,2 h and 24 h group HKC cells.The result of Oil Red O staining showed that lipid droplet markedly deposited in 6 h group HKC cells,contrarily,no lipid droplet was found in HKC cells of other groups.ConclusionAbove results suggested that insulin up-regulated SREBP-1 and FAS in time-dependent manner that led to cellular lipid droplet deposit,which may play an important role in the pathogenesis of renal lipid accumulation in metabolism syndrome.

Aim To investigate the effects of irbesartan on Wnt/β-catenin signaling pathway in tubular epithelial-mesenchymal transition(EMT)in HKCs induced by high-glucose.Methods Human kidney proximal tubular epithelial cell line(HKCs)cultured in vitro was divided into four groups:normal-glucose group,mannitol control group,high-glucose group and high-glucose plus irbesartan group.Immunocytochemistry staining was used to observe the expression of β-catenin;the protein expression of Wnt4,β-catenin,E-cadherin and α-SMA was assessed by Western blot;Wnt4 and β-catenin mRNA were detected by reverse transcription-polymerase chain reaction(RT-PCR).Results Compared with normal-glucose and mannitol control group,both the protein and the mRNA of Wnt4 were up-regulated in HKCs stimulated by high-glucose.α-SMA expression significantly increased but E-cadherin decreased in HG group.The cytoplastic and nuclear fraction of β-catenin enhanced with highglucose stimulation.But no difference of the total protein and mRNA of β-catenin was observed between highglucose-treatment and control groups.Highglucose induced Wnt4 and β-catenin expression in a time-dependent manner,both peaking at 24 h.Irbesartan reduced the promotional effect of HG on Wnt4 and α-SMA expression,and nuclear translocation of β-catenin.HG-mediated inhibition of E-cadherin was also restored by irbesartan.Conclusion These data supported a functional role for Wnt/β-catenin signaling pathway during epithelial-mesenchymal transition of HKCs induced by high glucose and suggested that irbesartan might reverse tubular EMT by regulating activity of Wnt/β-catenin pathway.

Objective To explore the effect of clinical and pathological features on the incidence of Hyperuricemia (HUA) in renal glomerular disease.Methods A retrospective analysis was applied to review the clinical and pathological date collected from 3547 patients with renal glomerular disease.These patients were diagnosed as renal glomerular disease by renal biopsy from January 2007 to December 2011.Results (1) HUA incidence was 21.8％ (773/3547) in all of the patients,in which the incidence in secondary glomerular disease 27.2％ (240/882) was much higher than that in primary glomerular disease 20.7％ (552/2665),and the difference was significant (x2 =153.642,P ＜ 0.05).In primary glomerular disease,HUA incidence was the lowest in membranous nephropathy 14.4％ (96/665),while HUA incidence in lupus nephritis (LN) 45.3％(110/243) was the highest and small blood vessel infammation kidney damage 34.7％ (17/49) was the second in secondary glomerular disease.(2) With the increasing of glomerulosclerosis index,tubulointerstitial score,renal vascular lesions score and the stage of chronic kidney disease,HUA incidence increased (x2 =17.798-298.216,P =0.000).(3)Logistic regression analysis showed that high tubulointerstitial score,glomerulosclerosis index and renal dysfunction,male,overweight or obese,hypertension and hypertriglyceridemia were risk factors for hyperuricemia (OR:1.011-7.513,P ＜ 0.05).Conclusion The uric acid level is increased in nearly a quarter of patients with renal glomerular disease.Severe tubulointerstitial lesion,high glomerulosclerosis index,low glomerular filtration rate,male,overweight or obese,hypertension and hypertiglyceridemia were independent risk factors for HUA.

Objective:To investigate the effect of HMGB1 and TLR-4 in the renal injury of SLE.Methods:The level of HMGB1 in serum from 16 patients suffering from SLE without kidney damage,18 patient with lupus nephritis (LN),and 12 healthy individuals were measured by ELISA.The fresh PBMCs were isolated and the total RNA was extracted.Then the mRNA expression of HMGB1 was amplified by RT-PCR.Flow cytometric analysis(FCM) was performed to study cell surface markers and the expression of TLR-4.Results:RT-PCR and ELISA results showed that the expressions of mRNA and protein were higher in patients with LN than in SLE without kindey damage and healthy people.The expression of TLR-4 in CD14+ monocytes of patients with LN was higher either,while there were no significant changes in CD3+ T cells among LN,SLE and healthy control.Conclusion:PBMCs in patients with LN could synthesize and secrete HMGB1 initiatively,which are correlated with serum HMGB1 level.HMGB1 might play a role in autoimmunity of lupus nephritis partly by activation of monocytes through its binding to TLR-4.

Objective To investigate the correlation between NF-?B/COX-2 signal pathway and cell proliferation in diabetic nephropathy. Methods Uninephrectomized STZ-induced male Wister rats were used as animal model. Using immunohistochemistry to detect NF-?B and COX-2 protein expressions in diabetic kidneys at the 16th week. HKC were cultured separately in normal or high glucose medium for 24,48,72 h.The expression of NF-?B and COX-2 protein was detected by flow cytometry and the expression of PCNA was detected by immunocytochemical staining. Results 1 Volum of glomeruli, mesangial matrix, thickness of glomerular and tubular basement membrane increased in diabete group; 2 COX-2 were expressed in cytoplasm of tubules and glomeruli by immunohistochemistry. Compared with control group, the expression of COX-2 was higher; activated NF-?B expressed in nucli of both tubules and glomeruli, There was light stainings for in control group, while enhanced stainings were observed in DM, there was a positive correlation between NF-?B and COX-2.3 Compared with those in HKC cultured in the medium with normal level glucose, the stainings were strengthened for PCNA in HKC exposed to high glucose from 24 h. 4 By FCM, the expression of NF-?B and COX-2 in HKC cultured in high glucose medium was higherthan that in normal glucose medium; the expression of NF-?B and PCNA was positively correlated with the expression of COX-2. Conclusion Activating NF-?B and elevating the expression of COX-2 play an important role in regulating cell proliferation, which may be one of the injury mechanisms of the renal cells during diabetic nephropathy.

Objective: To study the antioxidation and effects of water soluble extract from Natto on hyperlipidemia. Methods: The model of experimental hyperlipidemia was induced by high cholesterol diet. 18 rabbits were divided into 3 groups : model group, natto extract group and control group. Results: The natto extract could reduce the blood total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (LDL-C), malon-dialdehyde (MDA), atherogenic index (AI) of experimental rabbits 39.88% , 44.54% , 48.84%, 48.25% and 70.20%, respectively, and increase high density lipoprotein cholesterol (HDL-C), superoxide dismutase(SOD) 75.81% and 38.32%, respectively. Natto extract could prevent effectively the fatty degeneration of liver cells observed by pathological sections. Conclution: Natto extract can efficiently prevent the formation of experimental hyperlipidemia.