ObjectiveTo investigate the mechanism of danshenol A （DA） pretreatment in alleviating myocardial ischemia-reperfusion injury （MIRI） by regulating cardiomyocyte ferroptosis by in vivo and in vitro experiments. MethodsA MIRI model was established in SD rats， and an in vitro oxygen-glucose deprivation/reoxygenation （OGD/R） model was constructed with H9C2 cells. Both models were treated with DA. H9C2 cells were allocated into blank， model （OGD/R）， DA， ferroptosis inducer （erastin）， and ferroptosis inhibitor （Fer-1） groups. Cell viability was assessed by the methyl thiazolyl tetrazolium （MTT） assay. Biochemical assays were performed to measure the superoxide dismutase （SOD）， malondialdehyde （MDA）， glutathione （GSH）， and ferrous ion （Fe²⁺） levels. Dihydroethidium （DHE） fluorescence assay was adopted to quantify the reactive oxygen species （ROS） level. Real-time PCR and Western blot were employed to quantify the mRNA and protein levels， respectively， of prostaglandin-endoperoxide synthase 2 （PTGS2）， glutathione peroxidase 4 （GPX4）， ferritin heavy chain 1 （FTH1）， and acyl-coA synthetase long-chain family 4 （ACSL4）. Sixty SPF-grade healthy male SD rats were randomly assigned to control， model （MIRI）， DA， erastin， and Fer-1 groups. Enzyme-linked immunosorbent assay （ELISA） was adopted to measure the serum levels of cardiac troponin I （cTnI）， lactate dehydrogenase （LDH）， and creatine kinase （CK）. Histopathological changes in the myocardial tissue were observed by hematoxylin-eosin （HE） staining. Cardiomyocyte apoptosis was detected by terminal deoxynucleotidyl transferase-mediated nick end labeling （TUNEL）. The effect of DA on cardiomyocyte ferroptosis were observed and analyzed by in vivo and in vitro experiments. ResultsIn vitro experiment： compared with the blank group， the OGD/R model group showed reduced cell viability， elevated levels of ROS， MDA， and Fe²⁺， up-regulated mRNA and protein levels of ACSL4， lowered levels of SOD and GSH， and down-regulated mRNA and protein levels of PTGS2， GPX4， and FTH1 （P<0.05，P<0.01）. The DA and Fer-1 groups exhibited consistent trends： cell viability， SOD and GSH levels， and the mRNA and protein levels of PTGS2， GPX4， and FTH1 were significantly restored， while the ROS， MDA， and Fe²⁺ levels， and the mRNA and protein levels of ACSL4 were reduced （P<0.05，P<0.01）. In vivo experiment： Compared with the control group， the MIRI model group showed elevated serum levels of cTnI， LDH， and CK， increased cardiomyocyte apoptosis rate， risen levels of ROS， MDA， and Fe²⁺， and up-regulated mRNA and protein levels of ACSL4. However， both DA and Fer-1 groups exhibited reductions in the indicators above （P<0.05）. Compared with the control group， the MIRI model group demonstrated reduced levels of SOD and GSH and down-regulated mRNA and protein levels of PTGS2， GPX4， and FTH1 （P<0.05）. In contrast， both DA and Fer-1 upregulated these indicators （P<0.05）， effectively reversing the trends in the model group. In addition， the MIRI model group showed swelling of cardiomyocytes， disarrangement of cardiac muscle fibers， and massive inflammatory cell infiltration， which were alleviated in the DA and Fer-1 groups. ConclusionDA alleviates MIRI by inhibiting ferroptosis and inflammation， demonstrating therapeutic potential in acute myocardial infarction.

ObjectiveTo investigate the mechanism of danshenol A （DA） pretreatment in alleviating myocardial ischemia-reperfusion injury （MIRI） by regulating cardiomyocyte ferroptosis by in vivo and in vitro experiments. MethodsA MIRI model was established in SD rats， and an in vitro oxygen-glucose deprivation/reoxygenation （OGD/R） model was constructed with H9C2 cells. Both models were treated with DA. H9C2 cells were allocated into blank， model （OGD/R）， DA， ferroptosis inducer （erastin）， and ferroptosis inhibitor （Fer-1） groups. Cell viability was assessed by the methyl thiazolyl tetrazolium （MTT） assay. Biochemical assays were performed to measure the superoxide dismutase （SOD）， malondialdehyde （MDA）， glutathione （GSH）， and ferrous ion （Fe²⁺） levels. Dihydroethidium （DHE） fluorescence assay was adopted to quantify the reactive oxygen species （ROS） level. Real-time PCR and Western blot were employed to quantify the mRNA and protein levels， respectively， of prostaglandin-endoperoxide synthase 2 （PTGS2）， glutathione peroxidase 4 （GPX4）， ferritin heavy chain 1 （FTH1）， and acyl-coA synthetase long-chain family 4 （ACSL4）. Sixty SPF-grade healthy male SD rats were randomly assigned to control， model （MIRI）， DA， erastin， and Fer-1 groups. Enzyme-linked immunosorbent assay （ELISA） was adopted to measure the serum levels of cardiac troponin I （cTnI）， lactate dehydrogenase （LDH）， and creatine kinase （CK）. Histopathological changes in the myocardial tissue were observed by hematoxylin-eosin （HE） staining. Cardiomyocyte apoptosis was detected by terminal deoxynucleotidyl transferase-mediated nick end labeling （TUNEL）. The effect of DA on cardiomyocyte ferroptosis were observed and analyzed by in vivo and in vitro experiments. ResultsIn vitro experiment： compared with the blank group， the OGD/R model group showed reduced cell viability， elevated levels of ROS， MDA， and Fe²⁺， up-regulated mRNA and protein levels of ACSL4， lowered levels of SOD and GSH， and down-regulated mRNA and protein levels of PTGS2， GPX4， and FTH1 （P<0.05，P<0.01）. The DA and Fer-1 groups exhibited consistent trends： cell viability， SOD and GSH levels， and the mRNA and protein levels of PTGS2， GPX4， and FTH1 were significantly restored， while the ROS， MDA， and Fe²⁺ levels， and the mRNA and protein levels of ACSL4 were reduced （P<0.05，P<0.01）. In vivo experiment： Compared with the control group， the MIRI model group showed elevated serum levels of cTnI， LDH， and CK， increased cardiomyocyte apoptosis rate， risen levels of ROS， MDA， and Fe²⁺， and up-regulated mRNA and protein levels of ACSL4. However， both DA and Fer-1 groups exhibited reductions in the indicators above （P<0.05）. Compared with the control group， the MIRI model group demonstrated reduced levels of SOD and GSH and down-regulated mRNA and protein levels of PTGS2， GPX4， and FTH1 （P<0.05）. In contrast， both DA and Fer-1 upregulated these indicators （P<0.05）， effectively reversing the trends in the model group. In addition， the MIRI model group showed swelling of cardiomyocytes， disarrangement of cardiac muscle fibers， and massive inflammatory cell infiltration， which were alleviated in the DA and Fer-1 groups. ConclusionDA alleviates MIRI by inhibiting ferroptosis and inflammation， demonstrating therapeutic potential in acute myocardial infarction.

AIM:The direct renin inhibitor aliskiren displays antihypertensive and antialbuminuric effects in humans and in animal models . Emerging evidence has shown that aliskiren localizes and persists in medullary collecting ducts even after treatment was discontinued . The purpose of the present study was to investigate whether aliskiren regulates renal aquaporin expression and improves urinary concen -trating defect induced by lithium .METHODS:The mice were either fed with normal chow or LiCl diet (40 mmol/kg dry food per day for first 4 days and 20 mmol/kg dry food per day for last 3 days ) for seven days .Some mice were intraperitoneally injected aliskiren ( 50 mg/kg BW per day in saline ) .RESULTS:Mice injected aliskiren developed decreased urine output and increased urine osmolal -ity when compared with controls .Aliskiren significantly increased protein abundance of AQP 2 and phosphorylated-S256 AQP2 in the kidney inner medulla .Immunohistochemistry and immunofluoresence showed increased apical and intracellular labeling of AQP 2 and pS256-AQP2 in collecting duct principal cells of kidneys in mice treated with aliskiren .Aliskiren treatment prevented urinary concen-trating defect in lithium-treated mice , and improved the downregulation of AQP 2 and pS256-AQP2 protein abundance in inner medulla of the kidney .In primary cultured rat inner medulla collecting duct cells , aliskiren dramatically increased AQP 2 protein abundance which was significantly inhibited either by PKA inhibitor H 89 or by adenylyl cyclase inhibitor MDL 12330, indicating an involvement of the cAMP signalling pathway in mediating aliskiren-induced increased AQP 2 expression .CONCLUSION: The direct renin inhibitor aliskiren upregulates AQP 2 protein expression in inner medullary collecting duct principal cells and prevents lithium -induced nephro-genic diabetes insipidus ( NDI) likely via PKA-cAMP pathways .

Aim To investigate the influence of Ramu-lus mori polysaccharides ( RMP) on blood glucose and anti-oxidative effect in streptozotocin ( STZ )-induced diabetic mice .Methods Diabetic mice were induced by intraperitoneal injection with 120 mg? kg -1 STZ and were randomly divided into the following 5 groups with 20 animals per group: model group , valsartan group ( 20 mg? kg -1 ) , low-, medium-, high-dose (0.3,0.6,1.2 g? kg -1 ) of RMP groups.Other 20 normal mice were treated as normal control group .The mice were administered orally for 90 d.On 45 d of ad-ministration , the 24 h urine was collected through met-abolic cages for urine protein detection .Pathological changes of kidney tissues were observed through HE staining .The serum levels of urea nitrogen ( BUN) and creatinine ( Cr ) were detected by automatic biochemical analyzer; and the manganese superoxide dismutase (Mn-SOD), catalase(CAT), malonaldehyde(MDA) and mitochondrial respiratory chain complex Ⅰ,Ⅲac-tivity of kidney tissues were also determined .ELISA method was used to detect ROS content in renal cortex . The SIRT1 , FOXO1 and NF-κB protein expressions were analyzed by Western blot .Results Compared with model group, the FBG, microalbuminuria, BUN and Cr were decreased by RMP medication ( P <0.05).The activities of Mn-SOD, CAT and mitochon-drial respiratory chain complex Ⅰ,Ⅲ in RMP groups were enhanced , while MDA and ROS levels were re-duced. Moreover, the expressions of SIRT1 and FOXO1 were up-regulated by RMP , the expression of NF-κB was down-regulated ( P<0.05) .Conclusion RMP exerts renal protective effect through up-regula-ting the expressions of SIRT1 and FOXO1 in renal cor-tex , which may relate to the improvement of anti-oxida-tive capability .

Objective To evaluate the value of contrast-enhanced ultrasonography (CEUS) in quantitative diagnosis of chronic kidney dysfunction(CKD) by comparing it with color Doppler imaging (CDFI). Methods Tirty-three cases (15 males and 18 females) of clinical confirmed CKD (stage Ⅲ～Ⅴ)were included. Forty-five healthy volunteers were performed as control group. CEUS and CDFI were performed on each patient. After intravenous bolus injection of 1ml SonoVue each side,CEUS of renal cortex blood perfusion was collected successively,and a time-intensity curve(TIC) was created with Philips iU22 system's QLAB software. Resistance index(RI) and peak systolic velocity(PSV) of renal partial arteries were also tested. Results Compared with normal kidney,CKD patients had delayed perfusion and decreased intensity. Changes of area under curve(AUC), derived peak intensity(DPI), slope rate of ascending curve(A)and time to peak(TTP) were statistically significant ( P ＜0. 05). Sensitivities of AUC,DPI,A and TTP in diagnosis of CKD (stage Ⅲ～Ⅴ ) were 91.2% ,84. 9% ,90.9% and 85.3%, their specificities were 95.4%,88.9% ,93.3% and 90.9%, their accuracies were 93.6%, 87. 2%, 92.3% and 88. 5%, respectively. The results of CEUS were better than RI in CDFI (sensitivity 70.4%, specificity 37. 8%, accuracy 52.2%).Conclusions CEUS can precisely display the hemodynamic change in CKD ( stage Ⅲ～Ⅴ ), and is more sensitive than CDFI.

Objective To evaluate the contrast-enhanced ultrasonography (CEUS) in quantitative diagnosis of chronic renal insufficiency. Methods Correlation of CEUS indexes with glomerular filtration rate (GFR) detected by 99mTc-DTPA renography was examined. Thirty-three cases of clinical chronic renal insufficiency were enrolled in the study. They were 15 males and 18 females with average age of (43.33±6.78) years. After intravenous bolus injection of 1 ml SonoVue,CEUS of renal cortex blood perfusion was performed successfully, and a time-intensity curve (TIC)was created with PHILIPS iU22 system's QLAB software. A 148 to 222 MBq dose of 99mTc-DTPA was injected as a bolus from antecubital vein. Renal scintigraphic images were collected immediately and GFR was obtained. Results The significant correlation coefficients between GFR and CEUS quantitative indexes were as follows: rAUC (area under curve)=0.886 (P＜0.05), rA (slope rate of ascending curve, A) =0.804(P＜0.05). However, rDPI (derived peak intensity, DPI)=0.021 (P＞0.05), rTTP (time to peak, TTP) =0.043 (P＞0.05), rα (slope rate of descending curve,α)=0.039 (P＞0.05). Conclusion CEUS can precisely display the hemodynamic change of chronic renal insufficiency, which is well correlated with GFR by 99mTc-DTPA renography.

Objective To investigate the relationship between hemodynamic parameters from quantitative analysis of contrast-enhanced ultrasonography (CEUS) on hepatocellular carcinoma (HCC) and pathological tumor differentiated grades. Methods Seventy-seven HCC lesions were examinated and off-line analyzed with dynamic images. Quantitative parameters such as slope of decrease to half of peak (SD), increased signal intensity (ISI), area under the curve (AUC) and blood flow coefficient (BF) were acquired, and the standardized values (sISI, sAUC and sBF) included the ratio of parameters from tumor to those from hepatic parenchyma. These quantitative parameters were correlated with tumor grades according to Edmonson criteria. Results There was significant difference (P＜0.05) of SD, AUC and BF, as well as standardized values (sISI, sAUC and sBF) between different grades of HCC. AUC, BF, sISI, sAUC and sBF were negative correlated with differentiated grades, respectively (P＜0.05). Well-differentiated HCC had significantly higher perfusion values than HCC of other grades (P＜0.05). Conclusion Quantitative analysis of CEUS can assess differentiation of HCC indirectly, and might reveal biological behavior of malignant tumors.

Objective To investigate the correlations between quantitative parameters of contrast-enhanced ultrasonography(CEUS) and histopathological stage of liver fibrosis.Methods Sixty cases of hepatic fibrosis diagnosed pathologically were examined with CEUS.The dynamic images of CEUS were analyzed off-line with a quantitative software.Region of interest (ROI) were drawn separately in hepatic artery(HA),liver parenchyma(PA),main portal vein(PV) and right hepatic vein(HV).The time intensity curve(TIC) and some quantitative parameters,including arrival time,increased signal intensity(ISI),peak intensity(PI) and area under curve(AUC) were acquired.Intrahepatic transit time of contrast agents were calculated as the arrival time difference between hepatic artery(HA) and HV(HA-HVTT),between PV and HV(PV-HVTT) and between PA and HV (PA-HVTT).All the parameters were compared with histopathologic stage.Results All cases were grouped into mild fibrosis (S0 and S1,25 cases),moderate fibrosis (S2 and S3,17 cases) and severe fibrosis (S4,18 cases) according pathologic stage.HA-HVTT,PV-HVTT and PA-HVTT decreased with the development of liver fibrosis.The parameters of ISI,PI and AUC obtained from TIC of portal vein reduced with the progression of fibrosis,too(P＜0.05).Receive operating characteristic(ROC) curve analysis showed that the area under curve of HA-HVTT,PV-HVTT and PA-HVTT were 0.808,0.848,and 0.823 (S＞S1),and 0.856,0.903,and 0.806 (S＞S3),respectively.Conclusions Quantitative parameters of CEUS reflect the stage of liver fibrosis mediately.The parameters of HA-HVTT,PV-HVTT and PA-HVTT are closely correlated with the stage of liver fibrosis.

ular perfusion in tumors objectively, which is potential in monitoring tumor vascular response to antiangiogenic therapy.

0.05). Conclusions CEUS can accurately display the perfusion changes of CIRD model of dogs,it's more sensitive than the BUN and SCr. DPI and TTP are the most sensitive quantitative indexes of CEUS.