Objective To investigate the impact of Astragaloside-IV (AS-IV) on the progression of myocardial infarction (MI) through macrophage-dependent mechanisms by regulating Snail1 lactylation and acetylation, as well as the transforming growth factor β (TGF-β) pathway. Methods Oxygen glucose deprivation (OGD) was used to establish an in vitro myocardial ischemia model in rat cardiomyocytes (H9c2), which were then treated with AS-IV. Cell viability was assessed using CCK-8, apoptosis was evaluated by flow cytometry, and LDH levels were measured to assess cellular damage. RAW246.7 cells were treated with LPS, and lactate levels in the supernatant were measured using ELISA, while expression of macrophage phenotype markers was evaluated using Western blot. RAW246.7 cell-conditioned medium (CM) was co-cultured with H9c2 cells to assess the protective effects of AS-IV on macrophage CM-mediated H9c2 damage. RAW246.7 cells were induced to differentiate into M1-like macrophages using LPS (100 ng/mL) + IFN-γ (20 ng/mL), and Snail1 was overexpressed in M1 macrophages. Transfected M1 macrophage CM was co-cultured with H9c2 cells to validate the mechanisms of AS-IV in MI. An MI rat model was established by ligation of the left anterior descending coronary artery (LAD), and was treated with AS-IV. Cardiac function, myocardial cell apoptosis, and cardiac tissue pathology were studied using echocardiography, TUNEL, and HE staining, respectively. Results Compared to the OGD group, AS-IV treatment promoted cell viability, reduced apoptosis and decreased LDH release. LPS upregulated lactate levels in the supernatant of RAW246.7 cell cultures and induced polarization of RAW246.7 cells to the M1 phenotype. AS-IV attenuated the damaging effects of RAW246.7 cell CM on H9c2 cells . Overexpression of Snail1 in M1 macrophages weakened the protective effects of AS-IV on H9c2 cells . In vivo study, results showed that, compared to the MI group, AS-IV treatment reduced lactate levels in the hearts of MI rats, improved cardiac function and myocardial injury and attenuated myocardial cell apoptosis. Conclusion AS-IV inhibits TGF-β pathway activation through the suppression of Snail1 lactylation and acetylation in a macrophage-dependent manner, thereby mitigating myocardial cell damage following MI.
Animals ; Myocardial Infarction/drug therapy* ; Rats ; Snail Family Transcription Factors/metabolism* ; Macrophages/cytology* ; Myocytes, Cardiac/metabolism* ; Triterpenes/pharmacology* ; Saponins/pharmacology* ; Acetylation/drug effects* ; Apoptosis/drug effects* ; Mice ; Cell Line ; RAW 264.7 Cells ; Transforming Growth Factor beta/metabolism*

Human naïve pluripotent stem cells (PSCs) hold great promise for embryonic development studies. Existing induction and culture strategies for these cells, heavily dependent on MEK inhibitors, lead to widespread DNA hypomethylation, aberrant imprinting loss, and genomic instability during extended culture. Here, employing high-content analysis alongside a bifluorescence reporter system indicative of human naïve pluripotency, we screened over 1,600 chemicals and identified seven promising candidates. From these, we developed four optimized media-LAY, LADY, LUDY, and LKPY-that effectively induce and sustain PSCs in the naïve state. Notably, cells reset or cultured in these media, especially in the LAY system, demonstrate improved genome-wide DNA methylation status closely resembling that of pre-implantation counterparts, with partially restored imprinting and significantly enhanced genomic stability. Overall, our study contributes advancements to naïve pluripotency induction and long-term maintenance, providing insights for further applications of naïve PSCs.
Humans ; DNA Methylation/drug effects* ; Genomic Instability ; Pluripotent Stem Cells/metabolism* ; Cell Culture Techniques/methods* ; Cells, Cultured

Objective To investigate the changes of regulatory T cells (Treg) in patients with hepatocellular carcinoma (HCC) after ultrasound- guided percutaneous cool- tip radiofrequency ablation (RFA), and to discuss its influence on the prognosis. Methods A total of 30 patients with HCC were enrolled in this study. The percentage of Treg in peripheral blood was estimated with flow cytometry before RFA and one, 4, 7 and 12 months after RFA. During the follow-up period, the therapeutic effects were evaluated by contrast enhanced sonography or contrast enhanced CT scanning. By using the methods of receiver operating characteristic (ROC) curve and Kaplan-Meier survival function, the correlation of Treg dynamic changes with the progression-free survival time was analyzed. Results One month after RTA, the tumor response (TR) rate in the 30 patients was 93.3% (28/30), the tumor progression (TP) rate was 6.67%(2/30). The percentage of Treg before RFA was (9.42 ± 1.16)%, which decreased to (6.55 ± 0.97)% one month after RFA, the difference was statistically significant (t = 15.325, P <0.001). Twelve months after RFA, TR rate became 33.3%(10/30), and TP rate became 66.7%(20/30). The preoperative percentage of Treg of TR group was (8.75 ± 0.72)%, which was significantly lower than that of TP group (9.76 ± 1.20)%, the difference was statistically significant (t=-2.448, P=0.021). ROC curves indicated that the optimal cut-off value of Treg nadir was 4.82%, the sensitivity was 90.0% and the specificity was 60.0%. The optimal cut-off time to reach Treg nadir was 5.5 months, the sensitivity was 70.0% and the specificity was 85.0%. Kaplan-Meier curve analysis showed that after RFA the progression-free survival rate (PFS) of patients with Treg nadir ≤ 4.82% was significantly higher than that of patients with Treg nadir>4.82%. PFS of patients with reaching Treg nadir≥5.5 months was significantly higher than that of patients with reaching Treg nadir<5.5 months. Log-rank test results were字2=5.207, P=0.023; 字2=22.079, P < 0.001, respectively. Conclusion Percutaneous cool-tip radiofrequency ablation can decrease the percentage of Treg cells. Besides, Treg nadir and the time reaching Treg nadir can reflect the prognosis of HCC patients after RFA to a certain extent.

Objective:Glutamine is the main oxidative fuel of the enterocyte which enters the enterocyte primarily via amino acid transporters.The aim of the test was to study the distributions and functions of glutamine transporters in IEC-6 cell line.Methods:The rat intestinal epithelial cell line(IEC-6) was incubated in vitro.The mRNA expression of different glutamine transporters,protein expression of system ASCT2,and the [3H]-L-glutamine uptake were measured.Results:The mRNA of system ASCT2,SN1,ATA1,LAT1,LAT2 was expressed and the protein expression of ASCT2 was also validated in IEC-6.In Na+-containing buffer,the velocity of Na+-dependent glutamine uptake was(164.07?37.94) fmol/(mg protein?10min).In Na+-free buffer,the velocity of glutamine uptake was(58.71?10.51)fmol/(mg protein?10min).With the saturate dosage of MeAIB,the velocity of glutamine uptake was(81.02 ?19.59) fmol/(mg protein?10min).Conclusion:There may be five kinds of glutamine transporters(ASCT2,SN1,ATA1,LAT1,and LAT2) in IEC-6 cell.The Na+-dependent transporter was the major contributor(64.22%) to glutamine total uptake in IEC-6.The contributions of system A and the remainder were 50.62% and 13.60%,respectively.The Na+-independent transporter was the lesser contributor(35.78%).

Objective: This study was designed to investigate the variations of endotoxin,glutamine and TNF-? concentration in septic rats. Methods: A total of 90 adult male SD rats were divided as follows: control group(normal),experimental group(CLP).They were killed on hour 6,12,24,48,72 after surgery.Plasma,liver,small intestine and skeletal muscle were collected to measure the concentrations of glutamine in plasma and tissues,the levels of endotoxin and TNF-? in plasma. Results: In septic rats,plasma concentrations of endotoxin and TNF-? increased on hour 6,significantly increased on hour 12,attained the peak on hour 24,and decreased evidently on hour 48 and 72.Glutamine concentration in plasma elevated on hour 6 and 12,elevated significantly on hour 24 and 48,and decreased on hour 72.Glutamine concentration in liver increased on hour 6～12,increased significantly on hour 24,and decreased significantly on hour 48 and 72.Glutamine concentration in small intestine and skeletal muscle decreased on hour 6 and 12,and decreased significantly after 24 hour. Conclusion: During the early stage of sepsis,the plasma levels of endotoxin and TNF-? are increased significantly,the glutamine concentration in plasma and liver is also increased significantly,however,it was decreased evidently in skeletal muscle and small intestine.