This study investigated the role of the nuclear factor of activated T cells c3 (NFATc3) in vascular smooth muscle cells (VSMCs) during aortic aneurysm and dissection (AAD) progression and the underlying molecular mechanisms. Cytoplasmic and nuclear NFATc3 levels were elevated in human and mouse AAD. VSMC-NFATc3 deletion reduced thoracic AAD (TAAD) and abdominal aortic aneurysm (AAA) progression in mice, contrary to VSMC-NFATc3 overexpression. VSMC-NFATc3 deletion reduced extracellular matrix (ECM) degradation and maintained the VSMC contractile phenotype. Nuclear NFATc3 targeted and transcriptionally upregulated matrix metalloproteinase 9 (MMP9) and MMP2, promoting ECM degradation and AAD development. NFATc3 promoted VSMC phenotypic switching by binding to eukaryotic elongation factor 2 (eEF2) and inhibiting its phosphorylation in the VSMC cytoplasm. Restoring eEF2 reversed the beneficial effects in VSMC-specific NFATc3-knockout mice. Cabamiquine-targets eEF2 and inhibits protein synthesis-inhibited AAD development and progression in VSMC-NFATc3-overexpressing mice. VSMC-NFATc3 promoted VSMC switch and ECM degradation while exacerbating AAD development, making it a novel potential therapeutic target for preventing and treating AAD.

The efficient production of L-glutamate is dependent on the product's rapid efflux, hence researchers have recently concentrated on artificially modifying its transport system and cell membrane wall structure. Considering the unique composition and structure of the cell wall of Corynebacterium glutamicum, we investigated the effects of CmpLs on L-glutamate synthesis and transport in SCgGC7, a constitutive L-glutamate efflux strain. First, the knockout strains of CmpLs were constructed, and it was confirmed that the deletion of CmpL1 and CmpL4 significantly improved the performance of L-glutamate producers. Next, temperature-sensitive L-glutamate fermentation with the CmpL1 and CmpL4 knockout strains were carried out in 5 L bioreactors, where the knockout strains showcased temperature-sensitive characteristics and enhanced capacities for L-glutamate production under high temperatures. Notably, the CmpL1 knockout strain outperformed the control strain in terms of L-glutamate production, showing production and yield increases of 69.2% and 55.3%, respectively. Finally, the intracellular and extracellular metabolites collected at the end of the fermentation process were analyzed. The modification of CmpLs greatly improved the L-glutamate excretion and metabolic flux for both L-glutamate production and transport. Additionally, the CmpL1 knockout strain showed decreased accumulation of downstream metabolites of L-glutamate and intermediate metabolites of tricarboxylic acid (TCA) cycle, which were consistent with its high L-glutamate biosynthesis capacity. In addition to offering an ideal target for improving the stability and performance of the industrial strains for L-glutamate production, the functional complementarity and redundancy of CmpLs provide a novel target and method for improving the transport of other metabolites by modification of the cell membrane and cell wall structures in C. glutamicum.
Corynebacterium glutamicum/genetics* ; Glutamic Acid/biosynthesis* ; Fermentation ; Metabolic Engineering ; Bacterial Proteins/metabolism* ; Bioreactors/microbiology* ; Gene Knockout Techniques

BACKGROUND: Gait impairment is closely related to quality of life in patients with Parkinson's disease (PD). This study aimed to explore alterations in brain microstructure in PD patients and healthy controls (HCs) and to identify the correlation of gait impairment in the ON and OFF states of patients with PD, respectively. METHODS: We enrolled 24 PD patients and 29 HCs from the Movement Disorders Program at Beijing Friendship Hospital Capital Medical University between 2019 and 2020. We acquired magnetic resonance imaging (MRI) scans and processed the diffusion kurtosis imaging (DKI) images. Preprocessing of diffusion-weighted data was performed with Mrtrix3 software, using a directional distribution function to track participants' main white matter fiber bundles. Demographic and clinical characteristics were recorded. Quantitative gait and clinical scales were used to assess the status of medication ON and OFF in PD patients. RESULTS: The axial kurtosis (AK), mean kurtosis (MK), and radial kurtosis (RK) of five specific white matter fiber tracts, the bilateral corticospinal tract, left superior longitudinal fasciculus, left anterior thalamic radiation, forceps minor, and forceps major were significantly higher in PD patients compared to HCs. Additionally, the MK values were negatively correlated with Timed Up and Go Test (TUG) scores in both the ON and OFF in PD patients. Within the PD group, higher AK, MK, and RK values, whether the patients were ON or OFF, were associated with better gait performance (i.e., higher velocity and stride length). CONCLUSIONS PD exhibits characteristic regional patterns of white matter microstructural degradation. Correlations between objective gait parameters and DKI values suggest that dopamine-responsive gait function depends on preserved white matter microstructure. DKI-based Tract-Based Spatial Statistics (TBSS) analysis may serve as a tool for evaluating PD-related motor impairments (e.g., gait impairment) and could yield potential neuroimaging biomarkers.
Humans ; Parkinson Disease/diagnostic imaging* ; White Matter/physiopathology* ; Male ; Female ; Middle Aged ; Aged ; Gait/physiology* ; Diffusion Magnetic Resonance Imaging/methods* ; Diffusion Tensor Imaging/methods*

Objective:To establish a normal data model of fetal right ventricular size and function using echocardiography，and to explore the clinical value of quantitative assessment of right ventricular size and function in the diagnosis of congenital heart diseases.Methods:（1）A simple random sampling method was employed to collect 1 004 pregnant women with normal singleton pregnancies at 24 to 32 +6 weeks of gestation who underwent fetal cardiac ultrasound examinations at the First Affiliated Hospital of Air Force Medical University from January 2021 to December 2023. Two-dimensional and M-mode echocardiography were used to measure the right ventricular end-diastolic diameter（RVEDD），right ventricular end-diastolic area（RVEDA），tricuspid annular plane systolic excursion（TAPSE）during systole，and the right ventricular fractional area change（RVFAC）was calculated. The correlations between the above parameters and ultrasound gestational age（USGA）were analyzed. Moreover，percentile growth curves for each parameter were plotted. With the above parameters as dependent variables and the USGA as the independent variable，a Z-score model was established through regression analysis.（2）A stratified sampling method was adopted to select 30 fetuses diagnosed with hypoplastic right heart syndrome（HRHS）and 30 fetuses diagnosed with pulmonary stenosis（PS）during the same period as the case group. The model was verified，and the morphological and functional characteristics of the right ventricle were analyzed. Results:The data of RVEDD，RVEDA，TAPSE，and RVFAC in normal fetuses showed a skewed distribution. Each parameter showed good linear correlations with USGA（ r=0.836，0.834，0.846，0.242；all P<0.001）. The constructed percentile curves for each parameter indicated that RVEDD，RVEDA and TAPSE increased significantly with the growth of USGA，while RVFAC showed a slow downward trend. All parameters in the HRHS group and TAPSE and RVFAC in the PS group deviated significantly from the normal reference range（all P<0.001）. Conclusions:By analyzing RVEDD，RVEDA，TAPSE and RVFAC of normal fetuses，the percentile and Z-score normal reference value models of multiple parameters of fetal right ventricular size and function have been established，providing corresponding standards for quantitative analysis.

Aim To explore the predictive value of serum free triiodothyronine(FT3)on the long-term prognosis of patients with coronary heart disease after percutaneous coronary intervention(PCI).Methods All the subjects were from a prospective cohort study(PRACTICE study).In this study,15 250 patients with coronary heart disease after PCI in the First Affiliated Hospital of Xinjiang Medical University were selected,and the clinical data,FT3 and creatinine were collected.All the subjects were followed up regularly,and the primary follow-up endpoints were all-cause mortality and cardiogenic mortality,the secondary endpoints were major adverse cardiovascular events(MACE)and major adverse cardiovascular and cerebrovascular events(MACCE).According to the admission criteria,3 109 patients were finally in-cluded in this study.According to the baseline value of FT3,patients were divided into normal FT3 group(FT3:3.65～6.8 pmol/L,1 446 cases)and low FT3 group(FT3＜3.65 pmol/L,1 663 cases).Kaplan-Meier analysis was used for survival analysis,and Log-rank test was used for survival comparison.Multivariate Cox regression analysis was used to e-valuate the risk factors of the follow-up results of the two groups.Results Compared with the normal FT3 group,all-cause mortality and cardiogenic mortality in the low FT3 group increased significantly(P＜0.05).Kaplan-Meier analysis showed that the cumulative risk of all-cause mortality and cardiogenic mortality increased in the low FT3 group(P＜0.05).Multivariate Cox regression analysis indicated that the risk of all-cause mortality increased by 1.639 folds in the low FT3 group(HR=2.639,95％CI:1.385～5.348,P=0.007),while no statistical difference was found in cardiogenic mortality after adjusting for multiple factors(P=0.125).Conclusion The decrease in serum FT3 levels has important predictive value for all-cause mortality after PCI in patients with coronary heart disease.

Aim To investigate the role and mechanism of KLHL21 gene in myocardial infarction(MI)of mice.Methods KLHL21 gene knockout(KO)mice were generated using CRISPR/Cas9 technology,and C57BL/6 wild-type mice were used as controls.Sixty KLHL21 KO mice and 60 wild-type mice were randomly divided into four groups:WT+Sham group(n=30),WT+MI group(n=30),KO+Sham group(n=30)and KO+MI group(n=30).Postoperative is-chemic and infarct areas were assessed using TTC and Evans Blue staining,myocardial injury markers were measured by ELISA,cardiac function was evaluated by ultrasound,and histological changes were examined using HE and Masson stai-ning.Western blot was used to detect proteins related to the nuclear factor-κB(NF-κB)signaling pathway.Results KLHL21 protein expression in the myocardial tissue of KO mice was significantly lower than that in WT mice.The infarct area in KO+MI mice was significantly larger than that in WT+MI group.KO+MI mice showed reduced cardiac function compared with WT+MI mice.HE staining revealed myocardial cell loss,liquefactive necrosis,nuclear fragmentation,and significant neutrophil infiltration,while Masson staining showed aggravated fibrosis in KO+MI group.Serum tumor necrosis factor-alpha(TNF-α),interleukin-6(IL-6),creatine kinase-MB(CK-MB),and cardiac troponin Ⅰ(cTnⅠ)lev-els were significantly increased in KO+MI mice compared with WT+MI mice.Western blot analysis showed increased lev-els of phosphorylated inhibitor of nuclear factor-κB alpha(p-IKBα),P65,and P50,and decreased nuclear factor-κB al-pha(IKBα)in KO+MI mice.Conclusion KLHL21 gene plays a preventive role in myocardial infarction in mice,possibly through inhibition of NF-κB signaling pathway activation.

Aim To investigate the role and mechanism of KLHL21 gene in myocardial infarction(MI)of mice.Methods KLHL21 gene knockout(KO)mice were generated using CRISPR/Cas9 technology,and C57BL/6 wild-type mice were used as controls.Sixty KLHL21 KO mice and 60 wild-type mice were randomly divided into four groups:WT+Sham group(n=30),WT+MI group(n=30),KO+Sham group(n=30)and KO+MI group(n=30).Postoperative is-chemic and infarct areas were assessed using TTC and Evans Blue staining,myocardial injury markers were measured by ELISA,cardiac function was evaluated by ultrasound,and histological changes were examined using HE and Masson stai-ning.Western blot was used to detect proteins related to the nuclear factor-κB(NF-κB)signaling pathway.Results KLHL21 protein expression in the myocardial tissue of KO mice was significantly lower than that in WT mice.The infarct area in KO+MI mice was significantly larger than that in WT+MI group.KO+MI mice showed reduced cardiac function compared with WT+MI mice.HE staining revealed myocardial cell loss,liquefactive necrosis,nuclear fragmentation,and significant neutrophil infiltration,while Masson staining showed aggravated fibrosis in KO+MI group.Serum tumor necrosis factor-alpha(TNF-α),interleukin-6(IL-6),creatine kinase-MB(CK-MB),and cardiac troponin Ⅰ(cTnⅠ)lev-els were significantly increased in KO+MI mice compared with WT+MI mice.Western blot analysis showed increased lev-els of phosphorylated inhibitor of nuclear factor-κB alpha(p-IKBα),P65,and P50,and decreased nuclear factor-κB al-pha(IKBα)in KO+MI mice.Conclusion KLHL21 gene plays a preventive role in myocardial infarction in mice,possibly through inhibition of NF-κB signaling pathway activation.

Aim To explore the predictive value of serum free triiodothyronine(FT3)on the long-term prognosis of patients with coronary heart disease after percutaneous coronary intervention(PCI).Methods All the subjects were from a prospective cohort study(PRACTICE study).In this study,15 250 patients with coronary heart disease after PCI in the First Affiliated Hospital of Xinjiang Medical University were selected,and the clinical data,FT3 and creatinine were collected.All the subjects were followed up regularly,and the primary follow-up endpoints were all-cause mortality and cardiogenic mortality,the secondary endpoints were major adverse cardiovascular events(MACE)and major adverse cardiovascular and cerebrovascular events(MACCE).According to the admission criteria,3 109 patients were finally in-cluded in this study.According to the baseline value of FT3,patients were divided into normal FT3 group(FT3:3.65～6.8 pmol/L,1 446 cases)and low FT3 group(FT3＜3.65 pmol/L,1 663 cases).Kaplan-Meier analysis was used for survival analysis,and Log-rank test was used for survival comparison.Multivariate Cox regression analysis was used to e-valuate the risk factors of the follow-up results of the two groups.Results Compared with the normal FT3 group,all-cause mortality and cardiogenic mortality in the low FT3 group increased significantly(P＜0.05).Kaplan-Meier analysis showed that the cumulative risk of all-cause mortality and cardiogenic mortality increased in the low FT3 group(P＜0.05).Multivariate Cox regression analysis indicated that the risk of all-cause mortality increased by 1.639 folds in the low FT3 group(HR=2.639,95％CI:1.385～5.348,P=0.007),while no statistical difference was found in cardiogenic mortality after adjusting for multiple factors(P=0.125).Conclusion The decrease in serum FT3 levels has important predictive value for all-cause mortality after PCI in patients with coronary heart disease.

Objective:To establish a normal data model of fetal right ventricular size and function using echocardiography，and to explore the clinical value of quantitative assessment of right ventricular size and function in the diagnosis of congenital heart diseases.Methods:（1）A simple random sampling method was employed to collect 1 004 pregnant women with normal singleton pregnancies at 24 to 32 +6 weeks of gestation who underwent fetal cardiac ultrasound examinations at the First Affiliated Hospital of Air Force Medical University from January 2021 to December 2023. Two-dimensional and M-mode echocardiography were used to measure the right ventricular end-diastolic diameter（RVEDD），right ventricular end-diastolic area（RVEDA），tricuspid annular plane systolic excursion（TAPSE）during systole，and the right ventricular fractional area change（RVFAC）was calculated. The correlations between the above parameters and ultrasound gestational age（USGA）were analyzed. Moreover，percentile growth curves for each parameter were plotted. With the above parameters as dependent variables and the USGA as the independent variable，a Z-score model was established through regression analysis.（2）A stratified sampling method was adopted to select 30 fetuses diagnosed with hypoplastic right heart syndrome（HRHS）and 30 fetuses diagnosed with pulmonary stenosis（PS）during the same period as the case group. The model was verified，and the morphological and functional characteristics of the right ventricle were analyzed. Results:The data of RVEDD，RVEDA，TAPSE，and RVFAC in normal fetuses showed a skewed distribution. Each parameter showed good linear correlations with USGA（ r=0.836，0.834，0.846，0.242；all P<0.001）. The constructed percentile curves for each parameter indicated that RVEDD，RVEDA and TAPSE increased significantly with the growth of USGA，while RVFAC showed a slow downward trend. All parameters in the HRHS group and TAPSE and RVFAC in the PS group deviated significantly from the normal reference range（all P<0.001）. Conclusions:By analyzing RVEDD，RVEDA，TAPSE and RVFAC of normal fetuses，the percentile and Z-score normal reference value models of multiple parameters of fetal right ventricular size and function have been established，providing corresponding standards for quantitative analysis.

Background: Neuroblastoma (NB) is a malignant pediatric tumor requiring new therapies. Accumulating evidence has confirmed that micro RNAs play critical roles in NB metastasis. Dihydroartemisinin (DHA) is capable of inhibiting the growth of NB cells. The primary objective of the current investigation was to characterize a newly discovered microRNA, miR-32-5p, in terms of the functional role, underlying mechanism of action, and potential synergistic therapeutic impact in the context of NB metastasis. Materials and methods: Real-time quantitative polymerase chain reaction and Western blotting were employed to assess the expression levels of miR-32-5p and its target, vacuolar protein sorting 4B (VPS4B). Furthermore, Transwell assay was utilized to evaluate in vitro cell migration and invasion, whereas a metastasis xenograft model was established in nude mice via caudal vein injections. Results: Gene Expression Omnibus database and real-time quantitative polymerase chain reaction analysis showed that miR-32-5p was downregulated in human NB samples and NB cell lines, in comparison with the normal tissue and cell lines. Inhibiting miR-32-5p induced the migration and invasion of NB cells, whereas overexpression of miR-32-5p prevented the migration and invasion in NB cell lines. Furthermore, VPS4B was identified as the direct target of miR-32-5p and the miR-32-5p reduction associated with NB metastasis upregulated the expression of VPS4B. Conversely, overexpression of VPS4B reversed the suppressive effects ofmiR-32-5p onNB cells. Moreover, miR-32-5p increased the sensitivity to DHA both in NB cells and in the metastasis xenograft model of nude mice. Conclusions: The downregulation of miR-32-5p in NB regulates NB metastasis by targeting VPS4B. Moreover, miR-32-5b can improve the sensitivity of DHA in the xenograft mouse model. Our findings have important implications for the combined application of miR-32-5p and DHA in the treatment of NB.