Objective To investigate the effect of puerarin(Pue)on ferroptosis in myocardial cells of rats with diabetic cardiomyopathy(DCM)and to analyze its potential mechanisms.Methods A DCM rat model was established by high-fat,high-sugar diet combined with streptozotocin injection.The successfully modeled rats were randomly divided into DCM group,low-dose Pue group(125 mg/kg,gavage),high-dose Pue group(250 mg/kg,gavage)and valsartan group(30 mg/kg,gavage),with 10 rats in each group.Another 10 healthy SD rats were selected as blank control group.At the end of treatment,fasting blood glucose,tail artery blood pressure(BP)total cholesterol(TC),triglycerides(TG)and low-density lipoprotein cholesterol(LDL-C)levels were measured in all groups.The cardiac function related indicators[left ventricular ejection fraction(LVEF),left ventricular fractional shorten-ing(LVFS),left ventricular end-systolic diameter(LVDs)and left ventricular end-diastolic diameter(LVDd)]of rats in each group were recorded by ultrasonic apparatus.Histopathological changes in myocardial tissues were observed using hematoxylin-eosin(HE)staining and TUNEL staining.Levels of glutathione peroxidase(GSH-Px),superoxide dismutase(SOD),reactive oxygen species(ROS),malondialdehyde(MDA)and Fe2+were detected using enzyme-linked immunosorbent as-say(ELISA)kits.Western blotting was used to detect the expression levels of glutathione peroxi-dase 4(GPX4),long-chain acyl-CoA synthetase 4(ACSL4)and proteins involved in the protein kinase B/glycogen synthase kinase-3β(AKT/GSK-3β)signaling pathway.Results Compared with the blank control group,the DCM group exhibited disordered or fractured myocardial fibers and significant inflammatory cell infiltration.The apoptosis rate,LVDs,LVDd,blood glucose,BP,TC,TG,LDL-C,ROS,MDA,Fe2+and ACSL4 protein expression levels in the DCM group were signifi-cantly higher,and LVEF,LVFS,GSH-Px as well as SOD and the protein expression levels of GPX4,phosphorylated(p)-AKT,p-GSK-3β and p-PI3K were lower than those in the blank group(P＜0.05).Compared with the DCM group,the low-dose Pue group,high-dose Pue group and valsartan group showed reduced fractured myocardial fiber or disarray,more regular arrangement of myocardial fibers,and decreased inflammatory cell infiltration.The apoptosis rate,LVDs,LVDd,blood glucose,BP,TC,TG,LDL-C,ROS,MDA,Fe2+and ACSL4 protein expression levels in the low-dose Pue group,high-dose Pue group and valsartan group were significantly lower,and LVEF,LVFS,GSH-Px as well as SOD and the protein expression levels of GPX4,p-Akt,p-GSK-3 βand p-PI3K were significantly higher than those in the DCM group(P＜0.05),and the effect was more significant in the high-dose Pue group.Conclusion Pue treatment can effectively improve the metabolic level of DCM rats,inhibit the pathological damage and apoptosis of cardiomyocytes,alle-viate oxidative stress,and thereby improving the cardiac function of DCM rats.Its mechanism of ac-tion may be related to the inhibition of cardiomyocyte ferroptosis mediated by the AKT/GSK-3βpathway.

Aim To investigates whether sphingosine-1-phosphate(S1P)regulates the expression of mitochon-drial calcium uniporter(MCU)via the sphingosine-1-phosphate receptor/proline-rich tyrosine kinase 2(S1PR/Pyk2)sig-naling pathway,thereby reducing oxidative stress-induced mitochondrial damage and inhibiting mitochondria-related apopto-sis.Methods Human umbilical vein endothelial cells(HUVEC)were subjected to oxidative damage using hydrogen peroxide(H2O2)as a model.Different concentrations of S1P were applied to the oxidative damaged HUVEC.Addi-tionally,the S1PR1 agonist SEW2871,the S1PR1 inhibitor W146,and the Pyk2 inhibitor PF-562271 were used to explore the specific mechanism of S1P action.Results S1P treatment significantly alleviated oxidative damage in HUVEC and was accompanied by an increase in S1PR1 expression(P<0.05),while S1PR3 expression remained unchanged.Mean-while,the expression levels of Pyk2 and MCU decreased(P<0.05).SEW2871 further reduced mitochondrial damage,whereas W146 exacerbated it(P<0.05).Furthermore,the application of the Pyk2 inhibitor PF-562271 also reduced H2O2-induced mitochondrial damage(P<0.05),further confirming the role of Pyk2 in this process.Conclusion S1P reduces H2O2-induced mitochondrial damage and inhibits mitochondria-related apoptosis in HUVEC by suppressing Pyk2 expression via S1PR1.

Objective:To investigate impact of leonurine(Leo)on necrotic apoptosis in rats with myocardial ischemia/reperfu-sion(MIR)injury by regulating receptor interacting protein 1(RIP1)-receptor interacting protein 3(RIP3)-mixed lineage kinase domain-like protein(MLKL)pathway.Methods:Fifteen rats were randomly selected as sham surgery group(Sham),left anterior descending coronary artery of remaining rats were ligated to construct MIR model,and randomly grouped into MIR group,Leo group(15 mg/kg),Compound 6i group(1 mg/kg RIP1-RIP3-MLKL pathway activator Compound 6i)and Leo+Compound 6i group(15 mg/kg Leo+1 mg/kg Compound 6i),with 15 rats in each group.MIR group and Sham group were injected with equal amounts of physiological saline.Cardiac function indicators were observed through ultrasound electrocardiogram;ELISA was applied to detect myocardial injury indicators and inflammatory factor levels;HE staining was applied to observe pathological damage of heart;TTC staining was applied to detect myocardial infarction area;TUNEL staining was applied to detect cell apoptosis;Western blot was applied to detect expres-sions of RIP1-RIP3-MLKL pathway proteins.Results:Myocardial cells in Sham group were intact and arranged in an orderly manner;arrangement of myocardial cells was disordered and swollen,myofibril contracted,and sarcolemma was destroyed in MIR group,ejec-tion fraction(EF)and cardiac output(CO)level were obviously lower than Sham group(P<0.05),Mb,cTnⅠ,CK-Mb contents,IL-6,IL-18 levels,myocardial infarction area,myocardial cell apoptosis rate,RIP1,RIP3 and MLKL protein levels were obviously increased(P<0.05);compared with MIR group,Leo group had improved cell arrangement disorder and edema,and obviously increased EF and CO levels(P<0.05),Mb,cTnⅠ,CK-Mb contents,IL-6,IL-18 levels,myocardial infarction area,myocardial cell apoptosis rate,RIP1,RIP3 and MLKL protein levels were obviously reduced(P<0.05),trend of Compound 6i group was opposite;myocardial tissue structure of Leo+Compound 6i group was similar to MIR group,and Compound 6i eliminated cardioprotective effect of Leo on MIR rats.Conclusion:Leo may alleviate myocardial necrosis apoptosis in MIR rats by down-regulating RIP1-RIP3-MLKL pathway,thus playing a therapeutic role in MIR.

Aim To investigates whether sphingosine-1-phosphate(S1P)regulates the expression of mitochon-drial calcium uniporter(MCU)via the sphingosine-1-phosphate receptor/proline-rich tyrosine kinase 2(S1PR/Pyk2)sig-naling pathway,thereby reducing oxidative stress-induced mitochondrial damage and inhibiting mitochondria-related apopto-sis.Methods Human umbilical vein endothelial cells(HUVEC)were subjected to oxidative damage using hydrogen peroxide(H2O2)as a model.Different concentrations of S1P were applied to the oxidative damaged HUVEC.Addi-tionally,the S1PR1 agonist SEW2871,the S1PR1 inhibitor W146,and the Pyk2 inhibitor PF-562271 were used to explore the specific mechanism of S1P action.Results S1P treatment significantly alleviated oxidative damage in HUVEC and was accompanied by an increase in S1PR1 expression(P<0.05),while S1PR3 expression remained unchanged.Mean-while,the expression levels of Pyk2 and MCU decreased(P<0.05).SEW2871 further reduced mitochondrial damage,whereas W146 exacerbated it(P<0.05).Furthermore,the application of the Pyk2 inhibitor PF-562271 also reduced H2O2-induced mitochondrial damage(P<0.05),further confirming the role of Pyk2 in this process.Conclusion S1P reduces H2O2-induced mitochondrial damage and inhibits mitochondria-related apoptosis in HUVEC by suppressing Pyk2 expression via S1PR1.

Objective:To investigate impact of leonurine(Leo)on necrotic apoptosis in rats with myocardial ischemia/reperfu-sion(MIR)injury by regulating receptor interacting protein 1(RIP1)-receptor interacting protein 3(RIP3)-mixed lineage kinase domain-like protein(MLKL)pathway.Methods:Fifteen rats were randomly selected as sham surgery group(Sham),left anterior descending coronary artery of remaining rats were ligated to construct MIR model,and randomly grouped into MIR group,Leo group(15 mg/kg),Compound 6i group(1 mg/kg RIP1-RIP3-MLKL pathway activator Compound 6i)and Leo+Compound 6i group(15 mg/kg Leo+1 mg/kg Compound 6i),with 15 rats in each group.MIR group and Sham group were injected with equal amounts of physiological saline.Cardiac function indicators were observed through ultrasound electrocardiogram;ELISA was applied to detect myocardial injury indicators and inflammatory factor levels;HE staining was applied to observe pathological damage of heart;TTC staining was applied to detect myocardial infarction area;TUNEL staining was applied to detect cell apoptosis;Western blot was applied to detect expres-sions of RIP1-RIP3-MLKL pathway proteins.Results:Myocardial cells in Sham group were intact and arranged in an orderly manner;arrangement of myocardial cells was disordered and swollen,myofibril contracted,and sarcolemma was destroyed in MIR group,ejec-tion fraction(EF)and cardiac output(CO)level were obviously lower than Sham group(P<0.05),Mb,cTnⅠ,CK-Mb contents,IL-6,IL-18 levels,myocardial infarction area,myocardial cell apoptosis rate,RIP1,RIP3 and MLKL protein levels were obviously increased(P<0.05);compared with MIR group,Leo group had improved cell arrangement disorder and edema,and obviously increased EF and CO levels(P<0.05),Mb,cTnⅠ,CK-Mb contents,IL-6,IL-18 levels,myocardial infarction area,myocardial cell apoptosis rate,RIP1,RIP3 and MLKL protein levels were obviously reduced(P<0.05),trend of Compound 6i group was opposite;myocardial tissue structure of Leo+Compound 6i group was similar to MIR group,and Compound 6i eliminated cardioprotective effect of Leo on MIR rats.Conclusion:Leo may alleviate myocardial necrosis apoptosis in MIR rats by down-regulating RIP1-RIP3-MLKL pathway,thus playing a therapeutic role in MIR.

Damage to organelles plays a significant role in myocardial ischemia/reperfusion injury,which results in the dysfunction of mitochondria and other related organelles.The communication between mitochondria and other organ-elles can also affect the development of myocardial ischemia/reperfusion injury.For instance,the mitochondria-associated endoplasmic reticulum membrane provides a"seamless connection"and regulates the exchange of organelles and metabolites(such as ions,lipids and proteins)between the mitochondria and the endoplasmic reticulum,which subse-quently affects myocardial ischemia/reperfusion injury.However,there is a lack of studies regarding the interaction be-tween mitochondria and related organelles,which is a critical component in triggering myocardial ischemia/reperfusion inju-ry.Therefore,this article describes the role of mitochondrial crosstalk with endoplasmic reticulum,lysosomes and nuclei in myocardial ischemia/reperfusion injury,and aims to provide a theoretical basis for targeting mitochondrial crosstalk with other organelles in the treatment of myocardial ischemia/reperfusion injury.

Objective To determine the optimum dose of pentazocine when combined with propofol for gastroscopy in elderly patients.Methods One hundred and forty ASA physical status Ⅰ or Ⅱ patients,aged 6575 yr,scheduled for elective gastroscopy under general anesthesia,were randomly assigned into Ⅰ-Ⅳ groups (n =35 each) using a random number table.Before insertion of the gastroscope,pentazocine 0.2,0.4 and 0.8 mg/kg were injected intravenously in Ⅱ-Ⅳ groups,respectively,while the equal volume of normal saline was given in Ⅰ group.Propofol was then administered by target-controlled infusion (TCI).The half-effective concentration (EC50 of propofol was determined by up-and-down sequential trial.The target plasma concentration (Cp) was set at 3.5 μg/ml in the first patient.Gastroscopy was performed at 5 min after the target effect-site and plasma concentrations were balanced.The response to gastroscopy was defined as positive when body movement and/or bucking occurred during gastroscopy.Each time the Cp increased/decreased by 0.3 μg/ml in the next patient depending on whether or not the response to gastroscopy was positive.EC50 and 95 ％ confidence interval of propofol TCI inhibiting the response to gastroscopy were calculated using Probit analysis.The development of respiratory depression and hypotension was observed.Results EC50 (95 ％ confidence interval) of propofol TCI inhibiting the response to gastroscopy was 2.82 (2.63-3.02) μg/ml,2.78 (2.58-2.97) μg/ml,2.16 (2.00-2.32) μg/ml and 2.03 (1.88-2.19) μg/ml in Ⅰ-Ⅳ groups,respectively.Compared with group Ⅰ,EC50 was significantly decreased in Ⅲ and Ⅳ groups,and the incidence of respiratory depression was increased in Ⅳ group (P ＜ 0.05).Conclusion The optimum dose of pentazocine when combined with propofol is 0.4 mg/kg for gastroscopy in elderly patients.

AIM: Centrifuge training can improve forward acceleration (+Gz) endurance. This study was to analyzed the gene expression of rat heart affected by centrifuge, and to research the molecular mechanism of improving +Gz endurance by centrifuge training. METHODS: Differential expressed genes between high+Gz endurance (+16Gz) rats, of test group after trained 12 d and control were screened using suppression subtractive hybridization (SSH) and dot blot hybridization. The obtained expressed sequence tags (ESTs)were used as probes to perform RNA slot hybridization with heart total RNA isolated from each gruop of centrifuge training and high+Gz endurance and low+Gz endurance (+12Gz) rats, respectively. The positive ESTs were sequenced and analyzed using BLAST(nr) at NCBI.RESULTS: Three down-regulated ESTs were obtained from heart samples, all of them are new, and their expression levels were decreasing during centrifuge training. CONCLUSION: Centrifuge training can significantly affect the special gene expressions of rat heart, and the expression changes of these genes may be ralated to the mechainism that +Gz endurance can be improved by centrifuge training.

A new method to revel RFLPs is presented. The human genomic DNAwas purified by saturatedNaCl solution and the pAW101 probe labelled with digoxigenin-dUTP. The relationships of RFLPsand genetic patterns of PGM1 (phosphoglucomutase),EsD (esterase D),GLO1 (glyoxalase)and ACP(acid phosphatase ) between the fillal generation and parental generation were detected in 15 families(among them 11 cases were aborted fetuses). The probability of paternity (w)was caculated accor-ding to Essen - Moller's formula, each w vlua was over 99. 73 %, reached the standard of incladingpaternity. An effective,rapid, and non-toxic RFLPs technique was established, which is easy to man-age in common lab oratories.