Objective To explore the protective effects of pretreatment with the Mongolian medicine Jiruhen Gurigumu-7(JG-7)and Guangzao Sanwei Tang(GZ-3)on myocardial ischemia-reperfusion injury(MIRI)in mice.Methods 60 male C57BL/6J mice were randomly divided into sham operation(Sham)group,model(Model)group,compound danshen drip pill(CDDP)positive control group,JG-7 group,GZ-3 group,and 12 mice in each group to establish the MIRI model,and the H9C2 cells were randomly divided into Control(normoxic)group,H/R(hypoxia 6 h reoxygenation 14 h)group,H/R+JG-7 group,H/R+GZ-3 group.The mice in each group were tested for cardiac function indexes after 30 min of ischemia,24 h and 7 d of reperfusion,TTC staining to detect infarct area after 24 h of MIRI,HE staining to detect myocardial tissue structure and cellular morphology after 24 h of MIRI,TUNEL apoptosis kit to detect apoptosis of myocardial cells after 24 h of MIRI,Masson staining to detect myocardial fibrosis after 7 d of MIRI.Blood was taken from the abdominal aorta,serum was separated,and the indexes after oxidative stress of MIRI were detected in each group of mice,and the survival rate of H9C2 cells after H/R was detected in each group by CCK-8 method.Results The results of TTC showed that JG-7 and GZ-3 reduced the infarct area after 24 h of MIRI in mice.ELISA and kit assays proved that JG-7 and GZ-3 reduced creatine phosphokinase isoenzyme(Creatinekinase-MB,CK-MB),Lactic dehydrogenase(LDH),malondialdehyde(MDA)levels,and increased superoxide dismutase(SOD)levels.HE staining showed that JG-7 and GZ-3 improved myocardial pathology after MIRI 24 h.The results of TUNEL apoptosis assay showed that JG-7 and GZ-3 improved apoptosis in myocardial tissues 24 h after MIRI.Masson staining results showed that JG-7 and GZ-3 could reduce the area of myocardial tissue fibrosis after MIRI 7 d.CCK-8 assay results showed that JG-7 and GZ-3 could improve the cell survival rate after H/R in H9C2 cells.Conclusion Pre-treatment with Mongolian medicine Jiruhen Gurigumu-7 and Guangzao Sanwei Tang can reduce the damage caused after ischemia-reperfusion(I/R),decrease the area of myocardial infarction and fibrosis after I/R in mice,and protect the heart.

Objective To investigate the effect of naringenin(NAR)on right ventricular remodeling induced by hypoxic pulmonary hypertension(HPH)and its related mechanism.Methods SD rats were randomly divided into the control group(NC),the NC+NAR group,the HPH group and the HPH+NAR group,with 15 rats in each group.HPH model was established in a low-pressure hypoxic artificial chamber.After the successful construction of the model,rats in the NC+NAR group and the HPH+NAR group were given NAR 100 mg/(kg·d)gavage once a day for 4 weeks,while rats in the NC group and the HPH group were given the same volume of normal saline once a day for 4 weeks.Right ventricular free wall thickness(RVEDWT)and end-diastolic right ventricular diameter(RVEDD)were measured by echocardiography.Right ventricular systolic pressure(RVSP)and mean pulmonary artery pressure(mPAP)were measured by cardiac catheter.Right ventricular mass ratio(RV/BW)and right ventricular hypertrophy index(RVHI)were calculated.Masson and Sirius staining of right ventricle was used to calculate the collagen volume fraction(CVF).TUNEL staining was used to evaluate the apoptosis of right ventricular cardiomyocytes.The contents of malondialdehyde(MDA)and the activity of superoxide dismutase(SOD)in right ventricular myocardium were detected.The expression of Rho associated kinase(ROCK)protein was detected by Western blot assay.Results Compared with the NC group and the NC+NAR group,mPAP,RVSP,RVEDWT,RV/BW,RVHI,right ventricular CVF,right ventricular myocardial cell apoptosis rate,MDA content and ROCK1 and ROCK2 protein expression in right ventricular myocardial tissue were increased in the HPH group.RVEDD and SOD activity in right ventricular myocardium were decreased(P＜0.05).Compared with the HPH group,mPAP,RVSP,RVEDWT,RV/BW,RVHI,right ventricular CVF,right ventricular myocardial cell apoptosis rate,MDA content,ROCK1 and ROCK2 protein expression in right ventricular myocardial tissue were decreased in the HPH+NAR group,and RVEDD and SOD activity in right ventricular myocardium were increased(P＜0.05).Conclusion NAR can reduce HPH-induced right ventricular remodeling,and its mechanism may be related to inhibiting ROCK signaling pathway and further improving apoptosis and oxidative stress in right ventricular myocardium.

Objective:To establish a linear calibration method using two reference substances for seven characteristic peaks of Cassiae Semen under complicated chromatographic condition,and to optimize the method.Methods:Using 15 different types of screened chromatographic columns and 2 components as reference compounds pair,the linear calibration method with 2 reference substances was established to predict the retention time of the other 5 components,and the method was verified by unknown chromatographic columns and unknown samples.Combined with column confirmation number and average coincidence rate of target peaks,the location results were compared comprehen-sively,and the method was optimized according to the defect under the influence of complicated chromatographic condition.Results:The average conformity rate of the target peak of the method before optimization was 73.3％,and the average conformity rate of the target peak of the optimized method was 98.7％.The optimized method has a high-er average peak coincidence rate and a wider range of applicability for the chromatographic column.Conclusion:The optimized linear calibration method using two reference substances can assist the localization analysis of chromato-graphic peaks in the characteristic chromatogram under complicated chromatographic condition.

Objective To explore the protective effects of pretreatment with the Mongolian medicine Jiruhen Gurigumu-7(JG-7)and Guangzao Sanwei Tang(GZ-3)on myocardial ischemia-reperfusion injury(MIRI)in mice.Methods 60 male C57BL/6J mice were randomly divided into sham operation(Sham)group,model(Model)group,compound danshen drip pill(CDDP)positive control group,JG-7 group,GZ-3 group,and 12 mice in each group to establish the MIRI model,and the H9C2 cells were randomly divided into Control(normoxic)group,H/R(hypoxia 6 h reoxygenation 14 h)group,H/R+JG-7 group,H/R+GZ-3 group.The mice in each group were tested for cardiac function indexes after 30 min of ischemia,24 h and 7 d of reperfusion,TTC staining to detect infarct area after 24 h of MIRI,HE staining to detect myocardial tissue structure and cellular morphology after 24 h of MIRI,TUNEL apoptosis kit to detect apoptosis of myocardial cells after 24 h of MIRI,Masson staining to detect myocardial fibrosis after 7 d of MIRI.Blood was taken from the abdominal aorta,serum was separated,and the indexes after oxidative stress of MIRI were detected in each group of mice,and the survival rate of H9C2 cells after H/R was detected in each group by CCK-8 method.Results The results of TTC showed that JG-7 and GZ-3 reduced the infarct area after 24 h of MIRI in mice.ELISA and kit assays proved that JG-7 and GZ-3 reduced creatine phosphokinase isoenzyme(Creatinekinase-MB,CK-MB),Lactic dehydrogenase(LDH),malondialdehyde(MDA)levels,and increased superoxide dismutase(SOD)levels.HE staining showed that JG-7 and GZ-3 improved myocardial pathology after MIRI 24 h.The results of TUNEL apoptosis assay showed that JG-7 and GZ-3 improved apoptosis in myocardial tissues 24 h after MIRI.Masson staining results showed that JG-7 and GZ-3 could reduce the area of myocardial tissue fibrosis after MIRI 7 d.CCK-8 assay results showed that JG-7 and GZ-3 could improve the cell survival rate after H/R in H9C2 cells.Conclusion Pre-treatment with Mongolian medicine Jiruhen Gurigumu-7 and Guangzao Sanwei Tang can reduce the damage caused after ischemia-reperfusion(I/R),decrease the area of myocardial infarction and fibrosis after I/R in mice,and protect the heart.

[Objective] To conduct serological identification and molecular mechanism study on a ambiguous ABO blood group. [Methods] Standard serological techniques were used for the forward and reverse typing of ABO blood type. ABO gene coding and regulatory regions were analyzed by PCR after DNA extraction. Monoclonal sequencing was used to detect the haplotypes of the DNA sequence, and bioinformatics analysis was applied to predict the possible translation outcomes of the mutated DNA sequence. [Results] The sample’s red blood cells showed mixed field agglutination with anti-A, and the serum agglutinated with B cells, exhibiting serological characteristics of subtype A. Direct sequencing and monoclonal sequencing analysis of the ABO gene confirmed one allele as O02, the other had a c.27delC mutation compared with A102, which could cause the translation sequence to terminate prematurely at the 19th amino acids. Analysis and prediction suggested that the mutation might affect the function of the transferase through mechanisms such as shifting the initiation codon, altering the reading frame and affecting the splice sites. [Conclusion] This case is a rare A subtype caused by the c.27delC variation, and the impact on the glycosyltransferase may involve multiple mechanisms, which require further research and exploration.

OBJECTIVE To investigate the neuroprotective effects of Ditan Decoction(DTD)on ischemic stroke.METHODS A mouse middle cerebral artery occlusion(MCAO)model was used to induce cerebral ischemia and assess the role of DTD in post-stroke NVU injury.DTD was gavaged once a day for 3 days after MCAO.Transwell neutrophil chemotaxis assay was used to explore the role of DTD in the neutrophil chemotaxis.RESULTS In the MCAO model,DTD treatment significantly reduced infarct volume(P<0.01)and attenuated blood-brain barrier disruption,as evidenced by decreased IgG leakage and preserved laminin expression(P<0.05).Furthermore,DTD suppressed neutrophil infiltration into ischemic brain tissue,as demonstrated by reduced neutrophil elastase(P<0.01)and myeloperoxidase(P<0.05)levels.Mechanistically,DTD inhibited neutrophil chemotaxis in a dose-dependent manner and downregulated phosphodiesterase 4B(PDE4B),a key regulator of neutrophil migration(P<0.05).Molecular docking analysis i-dentified four active DTD components-apigenin,vitexin,chlorogenic acid,and orientin-with strong binding affinities to PDE4B(bind-ing energies<-5 kcal·mol-1),suggesting their potential role in mediating DTD's therapeutic effects.CONCLUSION These find-ings highlight DTD as a promising intervention for ischemic stroke,targeting NVU preservation and PDE4B-dependent neutrophil mod-ulation.

Objective:To investigate the mechanism by which microRNA-183 (miR-183) regulates the progression of diabetic nephropathy (DN) through targeting phosphatase and tensin homolog deleted on chromosome 10 (PTEN) and modulating the AKT signaling pathway, and to identify potential therapeutic targets for DN.Methods:(1) Bioinformatic analysis of miRNA expression: MiRNA expression datasets from diabetic nephropathy (DN) and control samples were obtained from the Gene Expression Omnibus database. Differential expression analysis was performed, and differentially expressed miRNAs (DEmiRNAs) were identified using thresholds of an absolute log 2 (fold changes) >1 and an adjusted P-value<0.05. The results were visualized in a volcano plot and a heatmap. (2) Animal model establishment and in vivo interventional studies: A DN rat model was induced by administration of a high-fat/high-sucrose diet combined with an intraperitoneal injection of streptozotocin. Rats were randomly assigned into four groups ( n=10 per group) using a random number table: control group, DN model group, miR-183 inhibitor negative control (NC) group, and miR-183 inhibitor group. The latter two groups received tail vein injections of the miR-183 inhibitor NC or the miR-183 inhibitor, respectively, for eight consecutive weeks. Parameters including fasting blood glucose, 24-hour urinary protein excretion, urinary albumin excretion rate (UAER), serum creatinine, and blood urea nitrogen (BUN) were measured. Renal histopathological changes were assessed by HE and PAS staining. Furthermore, the expression of candidate miRNAs from patient data was validated, and the mechanism of action of miR-183 was investigated using quantitative real-time PCR and Western blotting. (3) In vitro mechanistic investigations in cultured podocytes: Mouse podocyte clone-5 (MPC5) cells were cultured in vitro and subjected to the following conditions: normal glucose (5.3 mmol/L glucose), high glucose (30 mmol/L glucose), and osmotic control (5.3 mmol/L glucose+19.5 mmol/L mannitol). Cells in the logarithmic growth phase were transfected with the miR-183 inhibitor (100 nmol/L), miR-183 mimic (50 nmol/L), or their corresponding negative controls. A dual-luciferase reporter assay was conducted to validate the binding interaction between miR-183 and the 3'-untranslated region (3'UTR) of PTEN. The effects of miR-183 on the AKT signaling pathway, apoptosis-related proteins, and cell viability were evaluated by quantitative real-time PCR, Western blotting, and the cell counting kit-8 assay, respectively. Results:MiR-183 expression was markedly upregulated in renal tissues from DN patients and DN model rats (both P<0.05). Inhibition of miR-183 significantly reduced renal miR-183 levels by 90.2% ( P<0.01), decreased fasting blood glucose by 65.3% ( P<0.01), and improved renal function parameters, including reductions in urinary protein (40.3%), blood urea nitrogen (32.1%), urinary albumin excretion rate (22.5%), and serum creatinine (40.2%) (all P<0.01). Histological analyses showed attenuation of glomerular lesions and glycogen accumulation. Bioinformatic prediction and experimental validation identified PTEN as a direct target of miR-183, confirmed by dual-luciferase assays. In vitro, miR-183 inhibition increased PTEN expression, reduced AKT phosphorylation, promoted podocyte proliferation, and suppressed apoptosis (upregulation of Bcl-2 and downregulation of cleaved-caspase-3). These effects were abolished upon PTEN knockdown. Conclusions:miR-183 aggravates DN by targeting PTEN and activating the AKT signaling pathway. Inhibition of miR-183 improves renal function and reduces podocyte apoptosis, suggesting miR-183 as a potential therapeutic target for DN.

Acute carbon monoxide poisoning (ACMP) is one of the most common gas poisonings in the emergency department, with tens of thousands of people seeking medical attention for carbon monoxide (CO) poisoning each year. The severity of poisoning is dependent upon environmental and human factors, with hypoxia and oxidative stress being important mechanisms of cardiac toxicity induced by CO. Myocardial involvement is common in moderate to severe ACMP, including myocardial injury, myocardial infarction, arrhythmia, and sudden death, which are associated with a high risk of death. Ferroptosis is a cell death mechanism caused by iron-dependent lipid peroxidation (LPO), although ferroptosis has been shown to play a critical role in various cardiovascular diseases, the potential mechanism by which it contributes to ACMP-induced myocardial injury is unclear. This review discusses the established link between ferroptosis and cardiovascular disease and summarizes the potential role of ferroptosis in ACMP-induced myocardial injury and the detrimental effects of ACMP on the heart. Elucidating these mechanisms could guide the development of novel therapeutic strategies that target ferroptosis to mitigate ACMP-induced myocardial injury. This review aims to provide a theoretical foundation for future research on the potential use of ferroptosis as a therapeutic target for ACMP-induced myocardial injury.
Humans ; Carbon Monoxide Poisoning/complications* ; Ferroptosis ; Lipid Peroxidation ; Myocardium/pathology* ; Oxidative Stress

Objective:To investigate the mechanism by which microRNA-183 (miR-183) regulates the progression of diabetic nephropathy (DN) through targeting phosphatase and tensin homolog deleted on chromosome 10 (PTEN) and modulating the AKT signaling pathway, and to identify potential therapeutic targets for DN.Methods:(1) Bioinformatic analysis of miRNA expression: MiRNA expression datasets from diabetic nephropathy (DN) and control samples were obtained from the Gene Expression Omnibus database. Differential expression analysis was performed, and differentially expressed miRNAs (DEmiRNAs) were identified using thresholds of an absolute log 2 (fold changes) >1 and an adjusted P-value<0.05. The results were visualized in a volcano plot and a heatmap. (2) Animal model establishment and in vivo interventional studies: A DN rat model was induced by administration of a high-fat/high-sucrose diet combined with an intraperitoneal injection of streptozotocin. Rats were randomly assigned into four groups ( n=10 per group) using a random number table: control group, DN model group, miR-183 inhibitor negative control (NC) group, and miR-183 inhibitor group. The latter two groups received tail vein injections of the miR-183 inhibitor NC or the miR-183 inhibitor, respectively, for eight consecutive weeks. Parameters including fasting blood glucose, 24-hour urinary protein excretion, urinary albumin excretion rate (UAER), serum creatinine, and blood urea nitrogen (BUN) were measured. Renal histopathological changes were assessed by HE and PAS staining. Furthermore, the expression of candidate miRNAs from patient data was validated, and the mechanism of action of miR-183 was investigated using quantitative real-time PCR and Western blotting. (3) In vitro mechanistic investigations in cultured podocytes: Mouse podocyte clone-5 (MPC5) cells were cultured in vitro and subjected to the following conditions: normal glucose (5.3 mmol/L glucose), high glucose (30 mmol/L glucose), and osmotic control (5.3 mmol/L glucose+19.5 mmol/L mannitol). Cells in the logarithmic growth phase were transfected with the miR-183 inhibitor (100 nmol/L), miR-183 mimic (50 nmol/L), or their corresponding negative controls. A dual-luciferase reporter assay was conducted to validate the binding interaction between miR-183 and the 3'-untranslated region (3'UTR) of PTEN. The effects of miR-183 on the AKT signaling pathway, apoptosis-related proteins, and cell viability were evaluated by quantitative real-time PCR, Western blotting, and the cell counting kit-8 assay, respectively. Results:MiR-183 expression was markedly upregulated in renal tissues from DN patients and DN model rats (both P<0.05). Inhibition of miR-183 significantly reduced renal miR-183 levels by 90.2% ( P<0.01), decreased fasting blood glucose by 65.3% ( P<0.01), and improved renal function parameters, including reductions in urinary protein (40.3%), blood urea nitrogen (32.1%), urinary albumin excretion rate (22.5%), and serum creatinine (40.2%) (all P<0.01). Histological analyses showed attenuation of glomerular lesions and glycogen accumulation. Bioinformatic prediction and experimental validation identified PTEN as a direct target of miR-183, confirmed by dual-luciferase assays. In vitro, miR-183 inhibition increased PTEN expression, reduced AKT phosphorylation, promoted podocyte proliferation, and suppressed apoptosis (upregulation of Bcl-2 and downregulation of cleaved-caspase-3). These effects were abolished upon PTEN knockdown. Conclusions:miR-183 aggravates DN by targeting PTEN and activating the AKT signaling pathway. Inhibition of miR-183 improves renal function and reduces podocyte apoptosis, suggesting miR-183 as a potential therapeutic target for DN.