Roudoukou-8 San reduces the hypoxia/reoxygenation injury of cardiac myocytes through JAK2/STAT3 pathway / 中国药科大学学报

@#To study the protective effects of Roudoukou-8 San extract on hypoxia/reoxygenation injury of cardiac myocytes and its relationship with tyrosine protein kinase 2(JAK2)/signal transducer and activator 3(STAT3)signaling pathway, hypoxia/reoxygenation injury model of H9c2 cells was built by sodium dithionite(Na2S2O4). The vitality of the cells was tested by CCK-8; the contents of lactate dehydrogenase(LDH), creatine phosphate kinase(CK)and aspartate aminotransferase(AST)in cell culture medium were tested by fully automatic biochemical analyzer; the contents of catalase(CAT), superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px)in cells were tested by kits; cell apoptosis degree was tested by hoechst staining. And the protein expressions of JAK2, p-JAK2, STAT3, p-STAT3, Bcl-2 and Bax in myocardial cells of each group were tested by Western blot. Hypoxia for 1 hour and reoxygenation for 3 hours of 2 mmol/L Na2S2O4 caused damage of about 50% H9c2 cells. Compared with the model group, the extracts of Roudoukou-8 San with different concentrations could increase the viability of cells. Besides, contents of CK, LDH and AST in cell culture medium were decreased significantly, while contents of CAT, SOD and GSH-Px were increased significantly. At the same time, the expression of p-JAK2, p-STAT3 and Bcl-2 were significantly increased and that of Bax was significantly decreased. The effects of Roudoukou-8 San extract could bereduced by AG490 blocker. Therefore, Roudoukou-8 San extract possesses protective effects on Na2S2O4 induced-hypoxia/reoxygenation injury of cardiac myocytes through JAK2/STAT3 signaling pathway.

Effects of Roudoukou-8 San against hydrogen peroxide-induced injury of cardiomyocyte / 中国药科大学学报

@#To study the effects of Roudoukou-8 San against hydrogen peroxide-induced cardiomyocyte in neonatal rats and to explore its mechanism. Cardiomyocytes were isolated and cultivated by neonatal rats and the injure models were established by H2O2. Methyl thiazolyl tetrazolium(MTT)assay was used to detect the protective effects of Roudoukou-8 San on H2O2-induced cardiomyocyte. The effects of Roudoukou-8 San on myocardium morphology were observed under inverted microscope. The contents of lactate dehydrogenase(LDH), creatine kinase(CK)and aspartate aminotransferase(AST)in cell culture medium were determined by Automatic biochemical instrument; The levels of malonydialdehyde(MDA), superoxide dismutase(SOD)and NO in the cells were detected by kit method. The apoptotic morphology of cardiomyocytes was observed by Hoechst fluorescence staining. Cell apoptosis were measured by Annexin V and PI double staining and flow cytomety. 100μmol/L H2O2 for 2 h could cause about 50% of myocardial injury. In the inverted optical microscope, H2O2 model group showed increased cell gap, decreased cell count, cell cytoplasmic vacuoles and other obvious damage. Roudoukou-8 San protected cell from H2O2-induced morphlogical improved in different degrees, reduced the release of LDH, CK and AST content, reduced the content of MDA, NO in myocardial cells significantly and increased the activity of SOD. Roudoukou-8 San energy significantly inhibited H2O2 damage myocardial cell apoptosis. Our study suggested that Roudoukou-8 San can protect cardiomyocyte from H2O2-induced injury by improving the cell viability, reducing oxidative stress injury, inhibiting inflammatory reaction and inhibiting cell apoptosis.

Study on Protective Effect and Mechanism of Astragaloside Ⅳ on D-galactose-induced Primary Cardiomyocytes Apoptosis / 中国药房

OBJECTIVE:To study the protective effect and mechanism of astragaloside Ⅳ on D-galactose(D-gal)-induced primary cardiomyocytes apoptosis. METHODS:Wistar neonatal rat primary cardiomyocytes were isolated and cultured. The cardiomyocytes were divided into normal control group(DMEM high glucose medium without serum),D-gal group(DMEM high glucose medium without serum containing 5 g/L D-gal)and astragaloside Ⅳ low-concentration,medium-concentration and high-concentration groups(after cultured with DMEM high glucose medium without serum containing 25,50,100 μg/mL astragaloside Ⅳ for 1 h,and then replaced with DMEM high glucose medium without serum containing corresponding concentration of astragaloside Ⅳ and 5 g/L D-gal). Cardiomyocytes activity was detected by CCK-8 kit after cultured for 48 h.The apoptosis level was detected by Hoechst staining(cultured for 24 h)and flow cytometry(cultured for 48 h). The mRNA expressions of apoptosis related factors(Bcl-2,Bax,Caspase-3)and ANP in cardiomyocytes were detected by RT-PCR after cultured for 24 h. RESULTS:Compared with normal control group,the activity of cardiomyocytes,mRNA level of Bcl-2 and ratio of Bcl-2/Bax were decreased in D-gal group,while apoptosis rate and mRNA levels of Bax,Caspase-3 and ANP were increased, with statistical significance(P<0.05 or P<0.01). Compared with D-gal group,the activity of cardiomyocytes,mRNA level of Bcl-2 and ratio of Bcl-2/Bax were increased in astragaloside Ⅳ groups,while apoptosis rate and mRNA levels of Bax,Caspase-3 and ANP were decreased,with statistical significance(P<0.05 or P<0.01),and in concentration-dependent manner. CONCLUSIONS:Astragaloside Ⅳ can protect D-gal induced primary cardiomyocytes from apoptosis,the mechanism of which may be associated with the increase of Bcl-2/Bax ratio and the decrease of mRNA expressions of Caspase-3 and ANP.

Effects and mechanisms of shRNA interfered with expression of leucine-rich repeat containing G-protein coupled receptor 5 on the malignant behaviors of colorectal cancer stem cells / 中华消化外科杂志

Objective To investigate the effects and mechanisms of shRNA interfered with expression of leucine-rich repeat containing G-protein coupled receptor 5 (Lgr5) on the malignant behaviors of colorectal cancer stem cells (CSCs).Methods The experimental study was conducted.The CSCs expressing Lgr5+ were sorted by fluorescence activated cell sorting.Lgr5+ cells that were transfected with Lgr5-shRNA lentiviral vector and nontarget shRNA lentiviral vector were respectively allocated into the experimental group and control group.The percentage of Lgr5+ cells was analyzed by flow cytometery.The relative expression of Lgr5 mRNA was detected by fluorescence quantitative real-time polymerase chain reaction (qRT-PCR).The capacity of self-renewal was detected by sphere forming assay.The tumorigenesis in vitro and in vivo were respectively measured by colony formation assay and xenografting experiment.The mRNA expressions of stem cells related genes (Oct4,Sox2,Nanog,KLF4),CSCs genes (CD133,CD44,ALDH) and Wnt/β-catenin pathway key genes (Axin2,Wnt5a,Wnt3a,Fzd3,c-myc,VEGF,Ascl2,claudin-1) were detected by qRT-PCR.Measurement data with normal distribution were represented as-x±s.Comparison between groups was analyzed using the t test.Results (1)Transfection efficiency of shRNA lentiviral vector induced Lgr5 by flow cytometery was respectively 6.8％± 1.0％ in the experimental group and 92.7％±3.3％ in the control group,with a statistically significant difference (t =43.148,P＜0.05).The relative expression of Lgr5 mRNA measured by qPT-PCR was respectively 0.168±0.057 in the experimental group and 1.148±0.004 in the control group,with a statistically significant difference (t=28.778,P＜0.05).(2) The capacity of self-renewal was detected by sphere forming assay.The results of sphere forming assay:the number of spheres was 29±6 in the experimental group and 410± 10 in the control group,with a statistically significant difference (t =41.070,P＜0.05).The results of colony formation assay:the numbers of colonies in the experimental group and control group were respectively 72±4 and 412± 19,showing a statistically significant difference (t =31.433,P＜ 0.05).The results of tumorigenesis:the volumes of tumors in the experimental group and control group were respectively (81± 15)mm3 and (328±24)mm3,with a statistically significant difference (t=11.304,P＜0.05).(3) The effects of Lgr5 down-regulation on related genes,results of qRT-PCR detection:① The mRNA relative expressions of Oct4,Sox2,Nanog and KLF4 (stem cells related genes) were 0.377±0.093,0.662±0.104,3.591±0.300,0.425±0.091 in the experimental group and 1.957± 0.026,2.137±0.015,5.831±0.165,1.536±0.014 in the control group,with statistically significant differences (t=23.079,22.261,8.446,19.186,P＜0.05).② The mRNA relative expressions of CD133,CD44 and ALDH (CSCs genes) were 1.490±0.155,5.535±0.487,1.640±0.039 in the experimental group and 2.488± 0.061,9.908±0.332,5.718±0.292 in the control group,with statistically significant differences (t =8.170,9.667,27.849,P＜0.05).③The mRNA relative expressions of Axin2,Wnt5a,Wnt3a,Fzd3,c-myc,VEGF,Ascl2 and claudin-1 genes in the Wnt/β-catenin pathway were respectively 1.592±0.267,0.528±0.138,2.153±0.078,1.480±0.064,0.248±0.128,1.492±0.025,0.658±0.095,1.647±0.087 in the experimental group and 3.651±0.224,2.570±0.093,2.301±0.157,1.636±0.058,1.415±0.080,2.610±0.159,2.480±0.123,3.432±0.273 in the control group.There were statistically significant differences in the mRNA relative expressions of Axin2,Wnt5a,c-myc,VEGF,Ascl2 and claudin-1 genes between the 2 groups (t =7.316,15.332,12.649,12.320,14.831,9.063,P＜0.05),and no statistically significant difference in the mRNA relative expressions of Wnt3a and Fzd3 between the 2 groups (t =2.887,2.242,P＞0.05).Conclusion The malignant behaviors of colorectal CSCs are suppressed after shRNA lentivirus interfered with expression of Lrg5,and its mechanism is related to inhibiting activity of Wnt/β-catenin pathway.

Study on experimental autoimmune encephalomyelitis with oxymatrine on rats model / 中国生化药物杂志

Objective To establish experimental autoimmune cerebral spinal cord inflammation ( EAE) model rats, and observe the pathological changes and effect of oxymatrine on EAE model.Methods 30 rats were randomly divided into control group,EAE ( model group) group and oxymatrine group.The EAE symptom score was used to evaluate the rats after the model, and to observe the changes of its behavior.By HE staining and Kluver &Barrera myelin dyeing to observe the inflammation of the brain and spinal cord demyelinating changes.Results The animals in control group had no change in behavior and pathological.In model group, all animals occured behavioral changes, accompanied by varying degrees of demyelination and inflammatory infiltration of the brain and spinal cord.In oxymatrine group,6 rats did not appear EAE clinical manifestations and behavioral change, and the myelin structure was intact.Conclusion Oxymatrine can extend the incubation period experimental autoimmune encephalomyelitis in rats of the disease, relieve symptoms and protect nerve.

Pharmacodynamic studies on cardioprotective function of Zhurihen dripping pills in experimental animals / 中国生化药物杂志

Objective To study the cardioprotective function of Zhurihen dripping pills in experimental animals.Methods By hypoxia tolerance model and isoproterenol hydrochloride induced acute hypoxia model, measuring the time of death in mice.For rat serum LDH, CK, SOD and MDA were measured to observe protective effect of Zhurihen dripping pills on vasopressin-induced acute myocardial ischemia model.The effect of Zhurihen dripping pills on latency and durations were observed in acute arrhythmia induced by inhaled chloroform – epinephrine rabbits model.Results The hypoxia tolerance in mice and myocardial hypoxia tolerance significantly improved by Zhurihen dripping pill, and prolonged the survival time in mice.The serum level of LDH, CK, SOD and MDA significantly improved, and protected the impaired myocardial cell in rats.The latent period of arrhythmia was significantly prolonged and duration of arrhythmia was shortened in rabbits.Conclusion Zhurihen dropping pills an improve myocardial oxygen deficiency, anti-arrhythmia effect and protect myocardial cells.

Lipoprotein (a) and homocysteine potentiate the risk of coronary artery disease in male subjects / 中国医师进修杂志

Objective To discuss the rehtionship between lipoprotein [Lp (a)],homocysteine (Hcy) and coronary artery disease (CAD) in men.Methods Two hundred and twenty-two male patients admitted for coronary angiography were classified into CAD positive group (111 cases) and CAD negative group (111 cases).The clinical data and plasma Hcy and Lp(a) levels were recorded in both groups.Logistic regression analysis was used to analyze males CAD risk factors.Results Lp (a) ≥0.3 g/L was a risk factor for CAD (OR =5.04,95％ CI 1.88-13.51,P =0.001),whereas Hcy was not related to CAD (OR =1.35,95％ CI 0.63-2.89,P =0.443).However,when both factors were considered together in an interaction model,plasma high Hcy and high Lp (a) levels were risk factors for CAD (OR =11.54,95％ CI 2.62-45.61,P=0.003).Conclusion Plasma Lp (a) and Hcy levels may increase the incidence of male CAD process.

Comparison of two coils for MR angiography based on the SDNR method / 中国医疗器械杂志

In this paper, signal-to-noise ratio (SNR) and signal different-to-noise ratio (SDNR) methods were used to compare image quality using two different radiofrequency coils. The two coil types included an eight-element phased-array coil and a quadrature birdcage head coil with endcap and the comparison studies performed on a uniform cylindrical phantom and volunteer respectively. The results showed phased-array coil have advantages and proved effectiveness of the method in the evaluation and selection of coils.