Objective To evaluate the effect of methylene blue pertreatment on acute kidney injury induced by sepsis in rats.Methods Ninety male Sprague-Dawley rats,aged 1.5-2.5 months,weighing 200-250 g,were randomly assigned into 3 groups (n =30 each) using a random number table:sham operation group (S group),cecal ligation and puncture (CLP) group and methylene blue group (MB group).The rats were anesthetized with 10％ chloral hydrate 350 mg/kg.Normal saline 0.8 ml was injected via the caudal vein in and CLP groups,while 10％ methylene blue 15 mg/kg (in normal saline 0.8 ml) was injected via the caudal vein in group MB.Sepsis was induced by CLP after the end of administration in CLP and MB groups.Twenty animals in each group were chosen and observed for 72 h survival rate.Ten animals were sacrificed in each group at 18 h after operation and kidney specimens were removed for microscopic examination and for determination of the expression of poly (ADP-ribose) polymerase (PARP-1) using immunohistochemistry and Western blot.Blood samples were taken from the heart for determination of serum concentrations of blood urea nitrogen (BUN),creatinine (Cr),cystatin C and neutrophil gelatinase-associated lipocalin (NGAL).Results Compared with group S,the survival rate was significantly decreased at 24,48 and 72 h after operation,and the serum concentrations of BUN,Cr,cystatin C and NGAL and expression of PARP-1 in kidney tissues were increased in CLP and MB groups (P ＜ 0.05).Compared with group CLP,the survival rate was significantly increased at 24 and 48 h after operation,and the serum concentrations of BUN,Cr,cystatin C and NGAL and expression of PARP-1 in kidney tissues were decreased in group MB (P ＜ 0.05).The pathological changes were significantly attenuated in group MB as compared with group CLP.Conclusion Methylene blue pertreatment can attenuate acute kidney injury induced by sepsis in rats through down-regulating the expression of PARP-1.

Hepatic carcinoma is one of the most common malignant tumors in China.Autophagy activity and the change of Nrf2-ARE pathway play an important role in the process of liver tumors.Nrf2 which is an important regulator to liver cancer belongs to the CNC family.Research discovered that after the inhibition of autophagy,Nrf2-ARE pathway activation contributes to the progression of hepatocellular carcinoma.This article summarizes the relationship between autophagy and the Nrf2-ARE pathway and its impact on the hepatic carcinoma.

Objective To explore the influence of HepG2 cells' proliferation and autophagy by the Nrf2-ARE pathway,and provide the experimental basis for clinical exploring effective liver cancer treatment.Methods Hepatic carcinoma HepG2 cells were cultured,and its proliferation inhibition rates and the change of cell cycle' s in each phase were explored by the MTT assay and flow cytometry.The hepatoma cells' autophagy was qualitative observed by inverted phase contrast microscope and fluorescence microscope.Results Inhibitory rate of HepG2 cells was obviously higher in the Nrf2 inhibitor BML-111 group than control group (P ＜ 0.05),and the control group was aslo obviously higher than the Nrf2 inducer EGb group (P ＜ 0.05).Flow cytometric analysis showed that G1 phase cells in the cell cycle increased,S phase cells reduced and G2/M period cells relatively increased in the Nrf2 inhibitor BML-111 group.But G1 phase cells reduced,S phase cells increased and G2/M period cells relative reduced in the Nrf2 inducer EGb group.Inverted phase contrast microscope and fluorescence microscope checked that ranging from the size of the bubble and autophagosome formed in Hepatoma HepG2 cytoplasmic of the Nrf2 inhibitor BML-111 group.Conclusions The Nrf2-ARE pathway played an reverse inhibition on HepG2 cells' proliferation and autophagy.After the inhibition of Nrf2-ARE pathway,HepG2 cells mostly stayed in the G1 phase of the cell cycle.

The present study attempted to examine the effects of bile acid pool size on liver regeneration after hepatectomy. The rats were fed on 0.2% cholic acid (CA) or 2% cholestyramine for 7 days to induce a change in the bile acid size, and then a partial hepatectomy (PH) was performed. Rats fed on the normal diet served as the controls. Measurements were made on the rate of liver regeneration, the labeling indices of PCNA, the plasma total bile acids (TBA), and the mRNA expression of cholesterol 7alpha-hydroxylase (CYP7A1), farnesoid X receptor (FXR), and transcription factor c-Jun or c-fos. As compared with the normal and CA groups, the rate of liver regeneration was decreased on the day 3, and 7 after PH; the peak of the labeling indices of PCNA was delayed and the labeling indices were significantly reduced on the day 1; the TBA were also decreased on the day 1; the expression of FXR decreased but that of CYP7A1 increased at any given time; at the 1st, and 3rd h, the expression of c-Jun was declined in the cholestyramine group. The reduction in the bile acid pool size was found to delay the liver regeneration, which may be caused by the down-regulation of FXR and c-Jun expression.

AIM:To investigate the effect of neuropeptide Y(NPY) on intracellular free calcium([Ca2+]i) and Ca2+ sarcoplasmic reticulum of cardiomyocytes in rats.METHODS:Cardiomyocytes of neonatal Sprague-Dawley rats were incubated with NPY at concentration of 100 nmol/L for 24 h.Fluorescent indicator Fluo-4 AM was used to detect [Ca2+]i and Fluo-5N AM was used to detect Ca2+ in sarcoplasmic reticulum(SR).Calcium image was recorded by laser scanning confocal microscope.The SR Ca2+ load was estimated by caffeine-induced Ca2+ transient(CCT).RESULTS:24 h after incubation with NPY,compared with control group,the concentration of [Ca2+]i was significantly elevated(P

Objective To investigate the relationship between polymorphisms of N-acetyltransferase 2(NAT2)genes and anti-tuberculosis drug induced hepatic-injury(ADIH).Methods A 1:1 matched case-control study was conducted.One hundred and six cases fulfilling the criteria of ADIH were selected as ADIH group from the patients who received anti-tuberculosis therapy.whereas those patients without any hepatic inj ury related elinical symptoms during three months of follow-up period were selected as control.The genetic polymorphisms of the loci,NAT2481C/T,NAT2-590G/A and NAT2-857G/A,were determined by polymerase chain reaction and restriction fragment length polymorphism technique(PCR-RFLP)in patients who received antituberculosis therapy.The major environmental factors and genotypes were analyzed by univariate and multivariate conditional Logistic analyses.Results The T,AA allele frequencies of NAT2-481C/T,NAT2-590G/A and NAT2-857G/A were 7.5%,28.8%and 17.9%respectively in ADIH group,and 6.6%,18.9%and 17.5%,respectively in the control group.Univariate analysis demonstrated that the frequency of NAT2 slow acetylation genotype in ADIH group was significantly higher than that in control group with a crude OR(95%CI)of 2.250(1.140-4.441).Among 6 potential risk factors,i.e.education level,occupation,body mass index(BMI),smoking,drinking and the type of tuberculosis,the low BMI and drinking were two risk factors for ADIH.In multivariate analysis,ADIH remained associated with acetylation genotype after adjusting for BMI and drinking status.The adjusted OR(95%CI)was 2.246(1.086-4.644).Conclusion NAT2 slow acetylation genotype may be associated with the occurrence of ADIH.

AIM:To investigate the effects of microRNA-486-5p (miR-486-5p) on the senescence of human mesenchymal stem cells ( hMSCs).METHODS: The expression of miR-486-5p was determined by miRNA arrays and real-time PCR.By transfection of miR-486-5p mimic or inhibitor, up-regulation or down-regulation of miR-486-5p expres-sion in hMSCs was established.The effect of miR-486-5p and silence information regulator 1 (SIRT1) on hMSC telomerase activity and senescence were detected byβ-galactosidase staining.RESULTS:The expression of miR-486-5p was up-regu-lated in the old hMSCs compared with the young hMSCs.Up-regulation of miR-486-5p resulted in increasing senescence of hMSCs.Conversely, down-regulation of miR-486-5p resulted in decreasing cell senescence.The expression of SIRT1 and telomerase reverse transcriptase ( TERT) was down-regulated in the old hMSCs compared with the young hMSCs.Directly repression of SIRT1 expression inhibited the hMSC TERT protein expression and telomerase activity, but increased cell se-nescence.The regulation of miR-486-5p on hMSC senescence was attenuated by inhibiting the expression of miR-486-5p and SIRT1 together.CONCLUSION:miR-486-5p enhances senescence of hMSCs by decreasing the expression of SIRT1 and telomerase activity.

AIM: To investigate the effect of microRNA-708-5p (miR-708-5p) on the migration of human mesenchymal stem cells (hMSCs).METHODS:The expression of miR-708-5p was determined by miRNA arrays and re-al-time PCR.By transfection of miR-708-5p mimic or inhibitor, the up-regulation or down-regulation of miR-708-5p ex-pression in hMSCs was evaluated .The cell scratch and Transwell tests were used to detect the migration capability of hM-SCs.The effects of transmembrane protein 88 (TMEM88), a miR-708-5p target gene, onβ-catenin expression and migra-tion of hMSCs were detected .RESULTS:The expression of miR-708-5p was down-regulated in the old hMSCs compared with the young hMSCs.Up-regulation of miR-708-5p resulted in increasing migration of hMSCs.Conversely, down-regula-tion of miR-708-5p resulted in decreasing cell migration .The expression of TMEM88 was up-regulated in the old hMSCs compared with the young hMSCs , while the expression of β-catenin was down-regulated.Directly repression of TMEM88 expression increased the β-catenin expression and migration of hMSCs .The regulation of miR-708-5p on hMSCs was atten-uated by inhibiting the expression of miR-708-5p and TMEM88 together.CONCLUSION:miR-708-5p increases β-cate-nin expression and Wnt/β-catenin activity by repressing TMEM 88, thus enhancing the migration of hMSCs .

[ ABSTRACT] AIM:To investigate the effects of microRNA-378*( miR-378*) on the survival and apoptosis of human mesenchymal stem cells ( hMSCs ) .METHODS: The expression of miR-378* was determined by microRNA arrays and quantitative real-time PCR ( qRT-PCR) .H2 O2 was used to induce hMSCs apoptosis.By transfection of miR-378*mimic or inhibitor, we up-regulated or down-regulated miR-378* expression in hMSCs.The effect of miR-378*and connective tissue growth factor ( CTGF) on hMSC survival and apoptosis were detected by MTT, LDH, caspase-3/7 and TUNEL assays.RESULTS:The expression of miR-378*was up-regulated in the old hMSCs compared with the young hMSCs.H2 O2 increased the expression of miR-378*, decreased the expression of CTGF.Up-regulation of miR-378*re-sulted in increasing apoptosis and decreasing survival of hMSCs.Conversely, down-regulation of miR-378*resulted in de-creasing cell apoptosis and increasing survival.The regulation of miR-378*on hMSC apoptosis and survival was attenuated by inhibiting the expression of miR-378* and CTGF together.Direct repression of CTGF expression inhibited the hMSC survival and increased apoptosis.CONCLUSION:miR-378*enhances apoptosis of hMSCs by repressing the expression of CTGF.

The present study attempted to examine the effects of bile acid pool size on liver regeneration after hepatectomy.The rats were fed on 0.2% cholic acid(CA)or 2% cholestyramine for 7 days to induce a change in the bile acid size,and then a partial hepatectomy(PH)was performed.Rats fed on the normal diet served as the controls.Measurements were made on the rate of liver regeneration,the labeling indices of PCNA,the plasma total bile acids(TBA),and the mRNA expression of cholesterol 7alpha-hydroxylase(CYP7A1),farnesoid Ⅹ receptor(FXR),and transcription factor c-Jun or c-fos.As compared with the normal and CA groups,the rate of liver regeneration was decreased on the day 3,and 7 after PH; the peak of the labeling indices of PCNA was delayed and the labeling indices were significantly reduced on the day 1; the TBA were also decreased on the day 1; the expression of FXR decreased but that of CYP7A1 increased at any given time; at the 1st,and 3rd h,the expression of c-Jun was declined in the cholestyramine group.The reduction in the bile acid pool size was found to delay the liver regeneration,which may be caused by the down-regulation of FXR and c-Jun expression.