Objective to investigate the mapping and ablation strategy of premature ventricular contraction(PVC)originating from the free wall of tricuspid annulus.Methods With the Carto3 three-dimensional electro-anatomical mapping system and long sheath supporting,the PVC originating from the free wall of the tricuspid annulus was mapped and ablated by inverted U-shaped catheter via the femoral vein access.Results In 19 patients with PVC originating from the free wall of the tricuspid annulus,mapping and ablation with an inverted U-shaped catheter under the free wall of tricuspid annulus was carried out.The treatment was immediately successful in all the 19 patients,and no complications occurred.During the 6-month follow-up period,one patient developed recurrence of PVC.Conclusion The PVC originating from the free wall of the tricuspid annulus can be roughly judged by the features of the electrocardiogram of the body surface before operation,and the mapping and ablation treatment by using inverted U-shaped catheter is technically-simple and clinically-safe with reliable therapeutic efficacy.

Public health emergencies pose severe challenges to the public health sector and emergency management work in hospitals.Enhancing the emergency management capabilities in general hospitals is of great significance in promoting high-quality development of hospitals,improving the government's public governance system,alleviating social panic,and other aspects.However,there are the practical dilemmas of insufficient monitoring and early warning,imperfect guarantee systems,and lack of technological innovation in emergency management in general hospitals.The emergency management capabilities in general hospitals can be improved through normalized monitoring and disposal,standing facility and material teams,information-based power systems,and standardization of technical support,further promoting the innovation and development of the emergency management system in general hospitals.

Objective:To explore the function of MDM2 and its relationship with p53 at the cellular level during H 2O 2 induced oxidative damage. Methods:MLE-12 HALI cell models were established using 0.5 mmol/L H 2O 2, and were divided into three groups: normal control group, H 2O 2 injury group, H 2O 2+MDM2 overexpressed group, and H 2O 2+MDM2 shRNA group. Infection of MLE-12 cells with adenovirus vector overexpressing and silencing MDM2; Using immunoprecipitation (Co-IP) to analyze the interaction between MDM2 and p53; Western blotting was used to detect the protein expression levels of MDM2, p53, Bcl-2, Bax, and cleared caspase-3 after HALI modeling; Measure the apoptosis rate of cells in each group. Results:After transcriptome sequencing,the p53 signaling pathway closely related to HALI. Compared with the normal group, the expression of MDM2 in the H 2O 2 injury group was lower ( P<0.05); Compared with the H 2O 2 injury group, overexpression of MDM2 resulted in a decrease in the apoptosis rate of MLE-12 cells ( P<0.05), a decrease in the expression levels of p53, Bax, and cleared caspase-3 proteins, and an upregulation of MDM2 and Bcl-2 protein expression ( P<0.05). Compared with the H 2O 2 injury group, when MDM2 was silenced, the cell apoptosis rate increased ( P<0.05), and the expression levels of p53, Bax, and cleared caspase-3 proteins were upregulated, while the expression levels of MDM2 and Bcl-2 proteins decreased ( P<0.05). Co-IP experiments showed that MDM2 binds to p53 protein. Conclusions:MDM2 can exert a protective effect on HALI by inhibiting MLE-12 cell apoptosis through the p53/Bcl-2/Bax axis.

Objectives:This study aimed to elucidate the incidence,clinical characteristics and prognosis of ST-elevation during radiofrequency catheter ablation for atrial fibrillation(AF). Methods:Consecutive patients who underwent radiofrequency catheter ablation for AF in Tongren Municipal People's Hospital,Qixingguan District People's Hospital of Bijie City and Guizhou Provincial People's Hospital from January 2021 to August 2023 were enrolled in this study.All patients underwent CARTO three-dimensional electroanatomical mapping and radiofrequency ablation via atrial septal approach under local anesthesia.The ST-elevation of electrocardiogram was analyzed.Follow-up was performed at 1,3,6,and 12 months after radiofrequency ablation.AF recurrence and duration,use of antiarrhythmic drugs,and incidence of death,thromboembolism,bleeding,and perioperative complications were evaluated. Results:ST-elevation was observed in 5 out of 798 patients(0.62%).ST-elevation occurred after transseptal puncture in three patients and during PentaRay multielectrode mapping in two patients.Blood pressure was significantly increased in three patients with transient ST-elevation(<20 min)and hemodynamic collapse occurred in two patients with persistent ST-elevation(>20 min).Catheter ablation of AF was completed in 4 patients,1 patient suffered severe hemodynamic disorders during radiofrequency catheter ablation,and the procedure was stopped immediately,this patient died from multiple organ system failure on the fifth day after failed radiofrequency catheter ablation,and the other 4 patients had no perioperative complications.The mean follow-up was(6±3)months,only 1 patient developed short atrial tachycardia,and the other patients had no recurrent atrial fibrillation and palpitation. Conclusions:Transient or persistent ST elevation can occur in patients during AF ablation.Early detection and rapid management are needed to prevent severe hemodynamic instability and cardiogenic death.

Objective To investigate the impact of exosomal(Exos)-miR-21-5p(miR-21)targeting S-phase kinase associated protein 2(SKP2)derived from Type Ⅱ alveolar epithelial cells(AEC-Ⅱ)on the patho-genesis of bronchopulmonary dysplasia(BPD).Methods A total of 60 SD rats aged 6～8 weeks were utilized in this study,with 30 of them subjected to extraction and culture through differential adherent centrifugation.Density gradient centrifugation was employed for the isolation of AEC-Ⅱ derived exosomes,while vesicles from AEC-Ⅱ me-dium were extracted using density gradient centrifugation.These isolates were subsequently confirmed by transmission electron microscopy and particle size analysis,and the targeting relationship between miR-21 and SKP2 was validated through dual-fluorescein reporter gene assay.The remaining 30 mice were combined in a male-to-female ratio of 3:1 to facilitate pregnancy testing.These neonatal mice were randomly assigned into four experimental groups:air control group(Con group),hyperoxia group(BPD+PBS group),hyperoxia-treated mice receiving exosomes(BPD+Exos-miR-21 group),and hyperoxia combined with exosome miR-21 inhibitor treatment group(BPD+Exos-AV-miR-21 group).Neonatal SD rats will be exposed to 85％oxygen to establish a BPD model.Follow-ing 14 days of high oxygen treatment,the expression levels of miR-21 in lung tissues and exosomes will be assessed using RT-qPCR.HE staining will be employed to observe pathological changes in lung tissue,while mean alveolar linear intercept(MLI)and radial alveolar count(RAC)will be calculated.Superoxide dismutase(SOD),malondialdehyde(MDA),and total antioxidant capacity(T-AOC)levels will be determined spectrophoto-metrically,whereas reactive oxygen species(ROS)levels will be measured via fluorescence spectrophotometry.Additionally,Western blot analysis will assess the expression levels of SKP2,NR2F2,and VEGF-A proteins.Results The results obtained from electron microscopy and particle size analysis demonstrated that the vesicle structure isolated from AEC-Ⅱ cells corresponded to exosomes.Moreover,there was a significant upregulation of miR-21 expression in exosomes(P＜0.01).Subsequently,the dual luciferase gene reporter assay con-firmed SKP2 as the target of miR-21.Comparative analysis revealed that compared to the Con group,both BPD+PBS and BPD+Exos-AV-miR-21 groups exhibited disordered lung tissue structure with enlarged and simpli-fied alveoli,increased levels of ROS,MDA,and MLI along with elevated expression of SKP2 protein(P＜0.01).Conversely,RAC,SOD,T-AOC levels were downregulated alongside miR-21 expression while NR2F2 and VEGF-A protein expressions decreased significantly(P＜0.01).In contrast to the BPD+PBS group,the number of alveoli without alveoli increased in the BPD+Exos-miR-21 group leading to improved degree of alveolar simplification accom-panied by reduced MLI,ROS,MDA levels as well as decreased SKP2 protein expression(P＜0.01).Addition-ally ROC,SOD,T-AOC,and miR-21 expressions were upregulated while NR2F2and VEGF-A expressions were increased(P＜0.01).Conclusions The exosomal miR-21 derived from AEC-Ⅱ may potentially target SKP2,thereby inhibiting oxidative stress and promoting alveolar development.Consequently,it can improve BPD by enhancing the protein expression of NR2F2 and VEGF-A.

Objective To investigate the impact of exosomal(Exos)-miR-21-5p(miR-21)targeting S-phase kinase associated protein 2(SKP2)derived from Type Ⅱ alveolar epithelial cells(AEC-Ⅱ)on the patho-genesis of bronchopulmonary dysplasia(BPD).Methods A total of 60 SD rats aged 6～8 weeks were utilized in this study,with 30 of them subjected to extraction and culture through differential adherent centrifugation.Density gradient centrifugation was employed for the isolation of AEC-Ⅱ derived exosomes,while vesicles from AEC-Ⅱ me-dium were extracted using density gradient centrifugation.These isolates were subsequently confirmed by transmission electron microscopy and particle size analysis,and the targeting relationship between miR-21 and SKP2 was validated through dual-fluorescein reporter gene assay.The remaining 30 mice were combined in a male-to-female ratio of 3:1 to facilitate pregnancy testing.These neonatal mice were randomly assigned into four experimental groups:air control group(Con group),hyperoxia group(BPD+PBS group),hyperoxia-treated mice receiving exosomes(BPD+Exos-miR-21 group),and hyperoxia combined with exosome miR-21 inhibitor treatment group(BPD+Exos-AV-miR-21 group).Neonatal SD rats will be exposed to 85％oxygen to establish a BPD model.Follow-ing 14 days of high oxygen treatment,the expression levels of miR-21 in lung tissues and exosomes will be assessed using RT-qPCR.HE staining will be employed to observe pathological changes in lung tissue,while mean alveolar linear intercept(MLI)and radial alveolar count(RAC)will be calculated.Superoxide dismutase(SOD),malondialdehyde(MDA),and total antioxidant capacity(T-AOC)levels will be determined spectrophoto-metrically,whereas reactive oxygen species(ROS)levels will be measured via fluorescence spectrophotometry.Additionally,Western blot analysis will assess the expression levels of SKP2,NR2F2,and VEGF-A proteins.Results The results obtained from electron microscopy and particle size analysis demonstrated that the vesicle structure isolated from AEC-Ⅱ cells corresponded to exosomes.Moreover,there was a significant upregulation of miR-21 expression in exosomes(P＜0.01).Subsequently,the dual luciferase gene reporter assay con-firmed SKP2 as the target of miR-21.Comparative analysis revealed that compared to the Con group,both BPD+PBS and BPD+Exos-AV-miR-21 groups exhibited disordered lung tissue structure with enlarged and simpli-fied alveoli,increased levels of ROS,MDA,and MLI along with elevated expression of SKP2 protein(P＜0.01).Conversely,RAC,SOD,T-AOC levels were downregulated alongside miR-21 expression while NR2F2 and VEGF-A protein expressions decreased significantly(P＜0.01).In contrast to the BPD+PBS group,the number of alveoli without alveoli increased in the BPD+Exos-miR-21 group leading to improved degree of alveolar simplification accom-panied by reduced MLI,ROS,MDA levels as well as decreased SKP2 protein expression(P＜0.01).Addition-ally ROC,SOD,T-AOC,and miR-21 expressions were upregulated while NR2F2and VEGF-A expressions were increased(P＜0.01).Conclusions The exosomal miR-21 derived from AEC-Ⅱ may potentially target SKP2,thereby inhibiting oxidative stress and promoting alveolar development.Consequently,it can improve BPD by enhancing the protein expression of NR2F2 and VEGF-A.

MEK is a canonical effector of mutant KRAS; however, MEK inhibitors fail to yield satisfactory clinical outcomes in KRAS-mutant cancers. Here, we identified mitochondrial oxidative phosphorylation (OXPHOS) induction as a profound metabolic alteration to confer KRAS-mutant non-small cell lung cancer (NSCLC) resistance to the clinical MEK inhibitor trametinib. Metabolic flux analysis demonstrated that pyruvate metabolism and fatty acid oxidation were markedly enhanced and coordinately powered the OXPHOS system in resistant cells after trametinib treatment, satisfying their energy demand and protecting them from apoptosis. As molecular events in this process, the pyruvate dehydrogenase complex (PDHc) and carnitine palmitoyl transferase IA (CPTIA), two rate-limiting enzymes that control the metabolic flux of pyruvate and palmitic acid to mitochondrial respiration were activated through phosphorylation and transcriptional regulation. Importantly, the co-administration of trametinib and IACS-010759, a clinical mitochondrial complex I inhibitor that blocks OXPHOS, significantly impeded tumor growth and prolonged mouse survival. Overall, our findings reveal that MEK inhibitor therapy creates a metabolic vulnerability in the mitochondria and further develop an effective combinatorial strategy to circumvent MEK inhibitors resistance in KRAS-driven NSCLC.

BACKGROUND: Advances in organoid culture technology have provided a greater understanding of disease pathogenesis, which has been rarely studied in sepsis before. We aim to establish a suitable organoids-based intestinal injury model for sepsis. METHODS: Stable passaged organoids were constructed and pre-treated with lipopolysaccharide (LPS) to mimic sepsis-induced intestinal injury. The LPS-induced sepsis model was used as a reference. We used quantitative real-time polymerase chain reaction to evaluate the RNA levels of inflammatory factors and antimicrobial peptides. Enzyme-linked immunosorbent assay was used to evaluate the protein levels, hematoxylin and eosin staining was used to evaluate the pathology of the small intestine of mice, and immunohistochemistry and immunofluorescence were used to evaluate the intestinal epithelial barrier function. Perkin Elmer Operetta™ was used to obtain high-resolution images of three-dimensional organoids. RESULTS: An LPS concentration >150 μg/mL after 24 h was identified to cause organoid growth restriction. The fluorescence intensity of zonula occludens-1 and occludins at LPS concentrations >100 μg/mL decreased significantly after 24 h. After LPS stimulation for 8 h, the RNA expression levels of interleukin (IL)-1α, tumor necrosis factor alpha, granulocyte-macrophage colony-stimulating factor, IL-6, and regenerating islet-derived protein 3 alpha, beta, and gamma increased. These results resembled those of intestinal epithelial layer alterations in a mouse sepsis model. For IL-10, the RNA expression level increased only when the LPS level >200 μg/mL for 24 h. CONCLUSIONS This study provides the primary intestinal in vitro model to study the effects of LPS-induced intestinal injury resembling sepsis. This model provides a platform for immune associated mechanism exploration and effective drug screening.
Mice ; Animals ; Lipopolysaccharides/toxicity* ; Sepsis ; Intestinal Diseases ; Tumor Necrosis Factor-alpha ; Disease Models, Animal ; Organoids ; RNA

Background::Gut-resident macrophages (gMacs) supplemented by monocytes-to-gMacs differentiation play a critical role in maintaining intestinal homeostasis. Activating transcription factor 4 (ATF4) is involved in immune cell differentiation. We therefore set out to investigate the role of ATF4-regulated monocytes-to-gMacs differentiation in sepsis-induced intestinal injury.Methods::Sepsis was induced in C57BL/6 wild type (WT) mice and Atf4-knockdown ( Atf4+/-) mice by cecal ligation and puncture or administration of lipopolysaccharide (LPS). Colon, peripheral blood mononuclear cells, sera, lung, liver, and mesenteric lymph nodes were collected for flow cytometry, hematoxylin and eosin staining, immunohistochemistry, quantitative reverse transcription polymerase chain reaction, and enzyme-linked immunosorbent assay, respectively. Results::CD64, CD11b, Ly6C, major histocompatibility complex-II (MHC-II), CX3CR1, Ly6G, and SSC were identified as optimal primary markers for detecting the process of monocytes-to-gMacs differentiation in the colon of WT mice. Monocytes-to-gMacs differentiation was impaired in the colon during sepsis and was associated with decreased expression of ATF4 in P1 (Ly6C hi monocytes), the precursor cells of gMacs. Atf4 knockdown exacerbated the impairment of monocytes-to-gMacs differentiation in response to LPS, resulting in a significant reduction of gMacs in the colon. Furthermore, compared with WT mice, Atf4+/- mice exhibited higher pathology scores, increased expression of inflammatory factor genes ( TNF-α, IL-1β), suppressed expression of CD31 and vascular endothelial-cadherin in the colon, and increased translocation of intestinal bacteria to lymph nodes and lungs following exposure to LPS. However, the aggravation of sepsis-induced intestinal injury resulting from Atf4 knockdown was not caused by the enhanced inflammatory effect of Ly6C hi monocytes and gMacs. Conclusion::ATF4, as a novel regulator of monocytes-to-gMacs differentiation, plays a critical role in protecting mice against sepsis-induced intestinal injury, suggesting that ATF4 might be a potential therapeutic target for sepsis treatment.

Objective:To investigate the clinic effect of r-MHT and hematoma aspiration on the traumatic intracerebral hematoma.Methods:89 cases with traumatic intracerebral hematoma were given hematoma aspiration,47 of them were given r-MHT and hematoma aspiration,the clinic effect on the 1st,7th,14th day after treatment were evaluated by NIHSS,the hematoma volume before treatment on the 1st,7th,14th day after treatment were counted by Dotian formula.Results:The effective rate of treatment group was 93.6％,which was significantly higher than that of the control group (P＜0.05).The NIHSS score of treatment group was significantly higher than that of the control group(P＜0.05) on the 1 st day,1st,2nd week after treatment (P＜0.05).Conclusion:r-MHT and hematoma aspiration couldn effectively reduce the brain damage,improve the patient's neurological fumction in treating traumatic intracerebral hematoma.