BACKGROUND: Renal tubulointerstitial fibrosis is mainly featured as the accumulation of extracellular matrix (ECM) in renal interstitium. The tubular epithelial-myofibroblast transdifferentiation (TEMT) is important to the pathogenesy of renal tubulointerstitial fibrosis. OBJECTIVE: To examine the effects of sodium ferulate (SF) on TEMT, and ECM main components such as collagen Ⅰ, collagen Ⅲ and fibronectin, in rat renal tubular epithelial cellsinduced by transforming growth factor-beta 1 (TGF- β1)- DESIGN: Randomized and controlled experimental study based on cells. SETTING: Department of Kidney in West China Hospital of Sichuan University. MATERIALS: Rat renal tubular epithelial cells (NRK-52E) originated from American Type Culture Collection (ATCC), were offered by the laboratory of Department of Nephrology in Australian Monash Medical Center. Cell strain used in this study was cultured at the 36th passage. SF white crystal with water solubility and more than 98.0% purify, was from Chengdu Hengda Pharmaceutical Co., Ltd. Different concentrations of SF (125,250, 500μreel/L) were designed in this study. Rabbit anti-rat α-smooth muscle actin (α -SMA) was produced by Wuhan Boster Company. Enzyme-linked immunosorbent assay (ELISA) kit was the produced of Shanghai Senxiong Science and Technology Co.,Ltd. Human recombinant TGF- β1 was produced by R&D Company. DNA Engine OpticonTM real-time fluorescence quantitative polymerase chain reaction apparatus was the product of MJ Research Company. METHODS: Rat renal tubular epithelial cells (NRK-52E) cultured in vitro were divided into five groups. Control group was added with serum-contained DMEM; TGF-β1-induced group was added with TGF-β1 at final concentration of 5 ng/L; SF at different concentrations groups were added with 125, 250, 500 μ mol/L SF and TGF- β1 at final concentration of 5 ng/L,respectively. MAIN OUTCOME MEASURES: The contrast phase microscope, real-time fluorescence quantitative polymerase chain reaction and ELISA method were used to detect TEMT of NRK52E cells induced by TGF-β1 and levels of collagen Ⅰ, collagen Ⅲ and fibronectin in the supernatant. RESULTS: Morphology of NRK52E cells: Compared with control group, TGF-β1 could induce the transdifferentiation of NRK52E cells, showing fibroblast-like in morphology after 3 days, which were previously the typical road stone-like epithelial cells. In three different concentration SF groups, the morphologic transformation stimulated by TGF-β1 could be partly ameliorated in a dose-dependent manner. Expression of α-SMA mRNA: Compared with control group, 5 ng/L TGF- β1 enhanced expression of α-SMA at 6 hours, and reached a peak at 72 hours; SF depressed the expression in a dose-dependent manner at 72 hours (P < 0.05). Changes of ECM: After induced by 5 ng/L TGF- β1 for 72 hours, the levels of collagen Ⅰ, collagen Ⅲ and fibronectin in the supernatant increased significantly (P < 0.05), whereas SF decreased these levels in a dose-dependent manner (P < 0.05). CONCLUSION: TGF- β1 induces the TEMT, and promotes the secretion of collagen Ⅰ, collagen Ⅲ and fibronectin. SF can inhibit TGF- β1-induced TEMT In a dose-dependent manner.

OBJECTIVETo discuss the changes in Wnt pathway inhibiting factors in esophageal precancerosis lesions induced by methyl benzyl nitrosamine (MBNA) and the effect of Gexia Zhuyu decoction.

METHODWistar rats were subcutaneously injected with MBNA (3.5 mg x kg(-1) for twice per week to establish the model. Since the 1st day after the model establishment, they were orally administered with Gexia Zhuyu decoction (16, 8 mg x kg(-1)). At the 10th week, esophageal tissues were collected to observe the pathological changes of esophageal mucosa, detect SFRP1, sFRP4, Axin1, Axin2 and GSK-3β mRNA levels.by fluorescent quantitation PCR analysis and β-catenin protein level by Western blotting.

RESULTBeing induced by MBNA, rats in the model group showed slight atypical hyperplasia in the histopathological examination. Compared with the normal group, Gexia Zhuyu decoction dose high and low groups showed no significant pathomorphological and histological changes. The model group showed lower gene transcription levels of esophageal tissues sFRP1, sFRP4, Axin1 and Axin2 (P < 0.05 or P < 0.01) and higher β-catenin protein expression level (P < 0.01) than the normal control group. The Gexia Zhuyu decoction low dose group showed higher gene transcription levels of esophageal tissues sFRP1, sFRP4, Axin1 and Axin2 (P < 0.05 or P < 0.01) and lower β-catenin protein expression level (P < 0.01) than the normal control group.

CONCLUSIONUp-regulated β-catenin protein level and down-regulated Wnt pathway could enhance Wnt pathway activity of MBNA-induced esophageal precancerous lesions. Gexia Zhuyu decoction could down-regulate the β-catenin protein level and up-regulate the transcription level of Wnt pathway inhibiting factors, but could not block MBNA-induced esophageal precancerosis lesions.

Animals ; Axin Protein ; genetics ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; Esophageal Diseases ; drug therapy ; genetics ; metabolism ; pathology ; Glycogen Synthase Kinase 3 ; genetics ; metabolism ; Glycogen Synthase Kinase 3 beta ; Humans ; Male ; Necrosis ; Nitrosamines ; adverse effects ; Proteins ; genetics ; metabolism ; Rats ; Rats, Wistar ; Wnt Proteins ; genetics ; metabolism ; Wnt Signaling Pathway ; drug effects

BACKGROUND: Connective tissue growth factor (CTGF) is a kind of factor that can mediate downstream action of transforming growth factor-β 1 (TGF- β 1). The upregulation of connective tissue growth factor expression plays an important role in pathological changes of renal interstitial fibrosis.OBJECTIVE: To explore the effect of Sodium Ferulate on the expression of CTGF mRNA and protein in rats with unilateral ureteral obstruction (UUO) and pathological changes of renal interstitial fibrosis, and to compare with Losartan.DESIGN: Randomized and controlled animal trial.SETTING: Department of Nephrology, West China Hospital of Sichuan University, and College of Public Health, Sichuan University.MATERIALS: Twenty-four healthy adult male SD rats were selected from the Experimental Animal Center of Sichuan University. Sodium Ferulate was provided by Sichuan Hengda Pharmacy Co, Ltd (No. 050302); rabbit anti-rat CTGF by Santa Cruz; Western blotting by BioRAD, USA; DNA Engine OpticonTM real-time fluorescent quantitation PCR device by MJ Research, USA.METHODS: The experiment was performed at Research Laboratory of Clinical Medicine (grade BSL-1), College of Public Health, Sichuan University from May to December 2006. Twenty-four healthy rats were randomly divided into 4 groups (n=6): UUO model group, Sodium Ferulate group, Losartan group, and sham-operation group. According to the previous protocol, UUO models were established in UUO model group, Sodium Ferulate group, and Losartan group, and the other rats were subjected to sham operation. From the first day after UUO, Sodium Ferulate group was intragastrically (i.g.)administrated with 150 mg/kg/d Sodium Ferulate; Losartan group was administrated ig. with 20 mg/kg/d. Losartan; UUO and sham operation groups were administrated i.g. with matching normal saline. All rats were executed 14 days after UUO to harvest partial renal tissues. All experimental procedure was accorded with animal ethical standards.MAIN OUTCOME MEASURES: The mRNA and protein expressions of CTGF were quantified by real-time PCR and Western blot. The pathological changes of renal interstitial tissues were observed by hematoxylin/eosin (HE) and Masson staining.RESULTS: Twenty-four rats were included in final analysis. Fourteen days after UUO, CTGF mRNA and protein expressions in UUO model group were significantly increased compared with sham operation group, but the expressions in Sodium Ferulate group were significantly lower than model group (P < 0.05). Compared with Losartan treated group, there was no significant difference (P > 0.05). HE and Masson staining showed inflammatory cell infiltration and tubular and interstitial changes as well as collagen deposition in renal interstitial tissues on day 14 after UUO. Sodium Ferulate obviously improved the renal pathological changes in UUO rats (P < 0.05), and the effect was similar to Losartan (P > 0.05).CONCLUSION: Sodium Ferulate inhibits UUO-induced renal interstitial fibrosis. This action, similar to the effect of Losartan, might be due to downregulation of CTGF expression.

Objective To observe the treatment of ~(99)Te-MDP on typeⅡcollagen induced arthritis (CIA)in rat,and the effect on the expression of synovial MMP-3 and TIMP-1 mRNA.To explore the mech anisms of the ~(99)Te-MDP in the treatment of rheumatoid arthritis.Methods The rats in which C1A(n=24)were divided into three group:the control group(n=8),~(99)Tc-MDP group(n=8)and Methotrexate group(n=8). Arthritis were evaluated by arthritis index and histopathological index and the expressions of MMP-3 and TIMP-1 mRNA in synovium were detected by RT-PCR.Results①The arthritis indexs of the control group, the methotrexate group,the ~(99)Tc-MDP group were increased with time.②The histopathological scnres of the control group were significantly higher than those of methotrexate group and ~(99)Tc-MDP group(P＜0.01).The histopathological scores of cartilage destruction and bone erosion of ~(99)Tc-MDP group were lower than those of methotrexate group(P＜0.05).③The levels of MMP-3 mRNA of the control group,~(99)Tc-MDP group, methotrexate group were notably higher than those of the control group(P＜0.01).The levels of control group was notably higher than that of the ~(99)Tc-MDP group(P＜0.05).There was not significant difference in all groups on the levels of TIMP-1 mRNA(P＞0.05).Conclusions ~(99)Tc-MDP can notably relieve the arthritis symdrome and retard the catilage damage and bone erosion of CIA in rats,and could significantly decrease the MMP-3 mRNA in the synovium.Which may be one of the therapeutic mechanism.~(99)Tc-MDP is better than methotrexate in retarding catilage and bone erosion and decreasing MMP-3 mRNA in CIA rats in a 3-week therapeutic intervention.

Objective To determine the utility of single energy CT value and spectrum energy curve in identifying different cause of diffuse spleen enlargement.Methods 43 patients confirmed by either surgical pathology,aspiration biopsy or clinical comprehensive diagnosis and follow-up were assessed,including lymphoma with spleen infiltration(lymphoma group,n=18) and cirrhotic splenomegaly(liver cirrhosis group,n=25).All patients underwent upper abdomen CT scans in GSI mode and the GSI data were transferred to the Workstation AW 4.6 to acquire single energy CT value(40-140 keV,10 keV's interval) and spectrum energy curve of the spleen on the venous phase.All single energy CT values and the slope of curves were comparatively analyzed through independent-samples t test.The diagnostic efficiency were evaluated by ROC analysis.Results Under 40-140 keV energy range,single energy CT values were significantly lower in the lymphoma group than in the liver cirrhosis group(all P＜0.05).The spectrum energy curve were both types of decreasing.Under 40-90 keV,100-140 keV energy range,the slop of curves in the lymphoma group(2.42 ± 0.70,0.27± 0.08) were also significantly lower than in the liver cirrhosis group (3.11 ± 0.62,0.34± 0.07),respectively(all P ＜0.05).When the slope of curve under 40-90 keV energy range was selected as a diagnostic indicator,the area under the curve(AUC) would reach 0.77.If threshold value of 1.39 was taken,the sensitivity and specificity would be equal to 86 ％ and 64 ％,respectively.Conclusion Single energy CT value and spectrum energy curve are helpful for differentiation of lymphoma with spleen infiltration from cirrhotic splenomegaly.

Objective To compare the diuretic efficacy of torasemide as a 2-hour continuous infusion and as a bolus injection of equal dose in patients with nephrotic syndrome,and to investigate a preferable administration mode of torasemide for these patients.Methods Twenty-three hospitalized patients were randomized to receive torasemide 20 mg or 40 mg per day by either 2-hour intravenous infusion or bolus injection,and interchanged after 48 hours of washout.Results Patients received torasemide by 2-hour intravenous infusion exhibited significantly higher daily urinary volume,chloride excretion,sodium excretion and fractional excretion of sodium (FENa) within 24 hours than those by bolus injection (P ＜ 0.05).Significantly lower bound-state torasemide excretion,higher ratio of urinary volume to torasemide excretion and a markedly larger area under the curve in the plasma concentrationtime profiles were also observed in the infusion group (P ＜ 0.05).Conclusion 2-hour continuous infusion delivers a better diuretic effect compared with a bolus injection of equal dose of torasemide in patients with nephrotic syndrome.

Objective To analyze alterations in electrocardiogram (ECG) of adenosine test in 99Tcm-MIBI myocardial perfusion imaging(MPI)SPECT study. Methods A total of 641 patients were in cluded in the study. The patients each underwent 99Tcm-MIBI MPI with adenosine test. The ECGs were taken before, during, and after adenosine infusion. Results In all, abnormal ECGs were found in 205(32.0%) patients. During adenosine infusion, 20.6%(132/641) of patients suffered from arrhythmia,29.5%(39/132) had atrial premature beats, 34. 1% (45/132) had premature ventricular beats, and 6. 1% (8/132) had sinoatrial block. In addition, 5.3% (7/132) had first-, 24.2% (32/132) had second-, and 0.8%(1/132) had third-degree atrioventricular block (AVB). After adenosine infusion, 4.4%( 28/641) of patients suffered from arrhythmia, 57.1% (16/28) had atrial premature beats, 39.3%(11/28) had premature ventricular beats, and 3.6% (1/28) had sinoatrial block. The perfusion images showed ischemia in 36 patients and infarction in 8 patients. Adenosine infusion was terminated in 39 patients (6. 1%) because of poorly tolerated side effects. However, no death or acute myocardial infarction occurred in the study. Conclusions Adenosine pharmacologic test for 99TcmMIBI MPI may result in relatively high incidence of arrhythmia in ECG monitoring.

Objective To explore the correlationships between microperfusion diffusion indexes derived from intravoxel incoherent motion(IVIM)and perfusion values measured by arterial spin labeling (ASL)in renal allograft. Methods A total of 76 renal allograft recipients and 26 age-matched volunteers (group 0)were included in this prospective study. All subjects were underwent conventional MRI, IVIM and ASL MRI which were performed in the oblique-sagittal plane. Seventy-six recipients were divided into two groups based on the estimated glomerular filtration rate(eGFR):recipients with good allograft function(group 1, eGFR≥ 60 ml · min-1 · 1.73m-2,n=44)and recipients with impaired allograft function(group 2, eGFR<60 ml · min-1 · 1.73m-2,n=32). Three IVIM indexes values, including true diffusion coefficient(ADCslow), pseudo-diffusion coef fi cient(ADCfast), perfusion fraction(PF), and one ASL index value of renal cortex(renal blood flow, RBF)were measured. One-way analysis of variance and the least significant difference were used to compare the different of each cortical index values among three groups. Correlations between the ADCslow, ADCfast, PF, RBF and eGFR as well as the correlation among the indexes were evaluated using Pearson correlation coefficients. Results For cortical ADCslow, ADCfast, PF and RBF values, allografts with good function and impaired function showed significantly differences compared healthy controls(all P<0.01). In allografts with good function, cortical ADCslow,ADCfast,PF showed no significantly differences compared with controls(all P>0.05), but RBF value was significantly lower(P<0.05). The ADCslow, ADCfast, PF and RBF values of renal cortex were significantly lower in allografts with impaired function compared to allografts with good function(all P<0.01). In renal allografts, there were significant positive correlations between cortical ADCslow, ADCfast, PF, RBF value and eGFR(r values were 0.604, 0.552, 0.579 and 0.673, all P<0.01). Cortical ADCfast and PF value exhibited a significant correlation with RBF for recipients(r values were 0.501 and 0.423, all P<0.01). Conclusion Cortical ADCfast and PF values derived from IVIM and RBF measured by ASL show a significant positive correlation in renal allografts.

Objective: To explore the role of ROCK1 in oxidized low-density lipoprotein (ox-LDL) induced podocyte injury and its possible mechanism. Methods: The conditionally immortalized mouse podocyte cells were cultured in vitro and exposed to 20 μg/ml ox-LDL for 24 h. Western blotting was used to analyze the expression level of p-MYPT, nephrin, LC3-Ⅱ, p62, p-ULK1 in groups of control, ox-LDL, ROCK1 siRNA with ox-LDL, wtROCK1 with ox-LDL. Podocytes were incubated with DiI labeled ox-LDL for 4 h and fluorescence microscope was used to analyze lipid distribution. Results: Compared with control group, ox-LDL increased cell cholesterol accumulation, activated ROCK along with decreased nephrin, LC3-Ⅱ(P<0.05), and increased p62, and p-ULK1 expression (P<0.05). Over-expression of ROCK1 significantly decreased the expression of nephrin and LC3-Ⅱ, but up-regulated the levels of p62, p-ULK1 and cell cholesterol accumulation in ox-LDL stimulated podocytes (P<0.05). In contrast, Inhibition of ROCK1 protected podocyte by improved lipophagy. Conclusion ROCK1 mediated disfunction of lipophagy contributes to the ox-LDL induced podocyte injury.