In the present study, a mouse model of coronary artery ligation was employed to evaluate the effects of Jiming Powder on mitophagy in the mouse model of myocardial infarction and elucidate its underlying mechanisms. A mouse model of myocardial infarction post heart failure was constructed by ligating the left anterior descending branch of the coronary artery. The therapeutic efficacy of Jiming Powder was assessed from multiple perspectives, including ultrasonographic imaging, hematoxylin-eosin(HE) staining, Masson staining, and serum cardiac enzyme profiling. Dihydroethidium(DHE) staining was employed to evaluate the oxidative stress levels in the hearts of mice from each group. Mitophagy levels were assessed by scanning electron microscopy and immunofluorescence co-localization. Western blot was employed to determine the levels of key proteins involved in mitophagy, including Bcl-2-interacting protein beclin 1(BECN1), sequestosome 1(SQSTM1), microtubule-associated protein 1 light chain 3 beta(LC3B), PTEN-induced putative kinase 1(PINK1), phospho-Parkinson disease protein(p-Parkin), and Parkinson disease protein(Parkin). The results demonstrated that compared with the model group, high and low doses of Jiming Powder significantly reduced the left ventricular internal diameter in systole(LVIDs) and left ventricular internal diameter in diastole(LVIDd) and markedly improved the left ventricular ejection fraction(LVEF) and left ventricular fractional shortening(LVFS), effectively improving the cardiac function in post-myocardial infarction mice. Jiming Powder effectively reduced the levels of myocardial injury markers such as creatine kinase(CK), creatine kinase isoenzyme(CK-MB), and lactate dehydrogenase(LDH), thereby protecting ischemic myocardium. HE staining revealed that Jiming Powder attenuated inflammatory cell infiltration after myocardial infarction. Masson staining indicated that Jiming Powder effectively inhibited ventricular remodeling. Western blot results showed that Jiming Powder activated the PINK1-Parkin pathway, up-regulated the protein level of BECN1, down-regulated the protein level of SQSTM1, and increased the LC3Ⅱ/LC3Ⅰ ratio to promote mitophagy. In conclusion, Jiming Powder exerts therapeutic effects on myocardial infarction by inhibiting ventricular remodeling. The findings pave the way for subsequent pharmacological studies on the active components of Jiming Powder.
Animals ; Myocardial Infarction/physiopathology* ; Mitophagy/drug effects* ; Mice ; Drugs, Chinese Herbal/administration & dosage* ; Protein Kinases/genetics* ; Male ; Ubiquitin-Protein Ligases/genetics* ; Humans ; Disease Models, Animal ; Mice, Inbred C57BL ; Signal Transduction/drug effects*

This study employed a mouse model of coronary artery ligation to assess the effect and mechanism of Jiming Powder on mitochondrial autophagy in mice with myocardial infarction. The mouse model of heart failure post-myocardial infarction was established by ligating the left anterior descending coronary artery. The pharmacological efficacy of Jiming Powder was evaluated through echocardiographic imaging, hematoxylin-eosin(HE) staining, and Masson staining. The levels of malondialdehyde(MDA), Fe~(2+), reduced glutathione(GSH), and superoxide dismutase(SOD) in heart tissues, as well as MDA immunofluorescence of heart tissues, were measured to assess lipid peroxidation and Fe~(2+) levels in the hearts of mice in different groups. Ferroptosis levels in the groups were evaluated using scanning electron microscopy and Prussian blue staining. Western blot analysis was conducted to detect the levels of key ferroptosis-related proteins, including nuclear factor erythroid 2-related factor 2(NRF2), ferritin heavy chain(FTH), glutathione peroxidase 4(GPX4), solute carrier family 7 member 11(SLC7A11), heme oxygenase 1(HO-1), and Kelch-like ECH-associated protein 1(KEAP1). The results showed that compared with the model group, both the high-and low-dose Jiming Powder groups exhibited significantly reduced left ventricular internal diameter in systole(LVIDs) and left ventricular internal diameter in diastole(LVIDd), while the left ventricular ejection fraction(EF) and left ventricular fractional shortening(FS) were significantly improved, effectively enhancing cardiac function in mice post-myocardial infarction. HE staining revealed that Jiming Powder attenuated myocardial inflammatory cell infiltration post-infarction, and Masson staining indicated that Jiming Powder effectively reduced fibrosis in the infarct margin area. Treatment with Jiming Powder reduced the levels of MDA and Fe~(2+), indicators of lipid peroxidation post-myocardial infarction, while increasing GSH and SOD levels, thus protecting ischemic myocardium. Western blot results demonstrated that Jiming Powder reduced KEAP1 protein accumulation, activated the NRF2/HO-1/GPX4 pathway, and up-regulated the protein expression of FTH and SLC7A11, exerting an inhibitory effect on ferroptosis. This study reveals that Jiming Powder exerts a therapeutic effect on myocardial infarction by inhibiting ferroptosis through the NRF2/HO-1/GPX4 pathway, providing a foundation for subsequent research on the pharmacological effects of Jiming Powder.
Animals ; Ferroptosis/drug effects* ; Myocardial Infarction/physiopathology* ; NF-E2-Related Factor 2/genetics* ; Mice ; Drugs, Chinese Herbal/administration & dosage* ; Male ; Heme Oxygenase-1/genetics* ; Phospholipid Hydroperoxide Glutathione Peroxidase/genetics* ; Humans ; Mice, Inbred C57BL ; Signal Transduction/drug effects* ; Disease Models, Animal

Hearing loss is the most prevalent disabling disease. Cochlear implantation（CI） serves as the primary intervention for severe to profound hearing loss. This consensus systematically explores the value of genetic diagnosis in the pre-operative assessment and efficacy prognosis for CI. Drawing upon domestic and international research and clinical experience, it proposes an evidence-based medicine three-tiered prognostic classification system（Favorable, Marginal, Poor）. The consensus focuses on common hereditary non-syndromic hearing loss（such as that caused by mutations in genes like GJB2, SLC26A4, OTOF, LOXHD1） and syndromic hereditary hearing loss（such as Jervell & Lange-Nielsen syndrome and Waardenburg syndrome）, which are closely associated with congenital hearing loss, analyzing the impact of their pathological mechanisms on CI outcomes. The consensus provides recommendations based on multiple round of expert discussion and voting. It emphasizes that genetic diagnosis can optimize patient selection, predict prognosis, guide post-operative rehabilitation, offer stratified management strategies for patients with different genotypes, and advance the application of precision medicine in the field of CI.
Humans ; Cochlear Implantation ; Prognosis ; Hearing Loss/surgery* ; Consensus ; Connexin 26 ; Mutation ; Sulfate Transporters ; Connexins/genetics*

Objective:To investigate the clinicopathological characteristics of ossifying fibromyxoid tumor (OFMT) with rare fusion subtypes.Methods:Three cases of OFMT with rare fusion subtypes, diagnosed and consulted in the Zhejiang Hospital, Zhejiang Provincial People′s Hospital, Hangzhou, China and Ningbo Clinical Pathology Diagnosis Center, Ningbo, China from January 2016 to December 2024 were collected. Immunohistochemistry (IHC), fluorescence in situ hybridization (FISH), and targeted RNA sequencing were performed to analyze the immunohistochemical and molecular genetic characteristics of these OFMT. Literature review was also conducted.Results:All three patients were male, with ages of 50, 74, and 58 years, respectively. The tumors were located in the left foot, left thigh, and left lumbar region, respectively, and all presented as slowly growing, painless masses in the skin or subcutaneous tissue. Grossly, the tumors measured 3.5 cm, 6.3 cm, and 5.0 cm in maximum diameter, respectively, with a grayish-white to grayish-yellow, solid, lobulated cut surface. One case exhibited a noticeable myxoid texture. Microscopically, one tumor was located in the superficial dermis, while the other two were in the subcutaneous tissue. The tumors were well-demarcated and showed a lobulated or multinodular growth pattern. None of the cases had a complete surrounding bony shell (only one case had very focal ossification). The tumor cells were monomorphic, short spindle-shaped, oval to epithelioid, and arranged in solid sheets, trabeculae, and small nests within a variably fibromyxoid stroma. Case 1 exhibited abundant pseudorosette-like structures formed by short spindle cells surrounding acellular fibrous stroma. Case 2 showed focal transition of epithelioid tumor cells into fasciculately arranged spindle cells, with extensive stromal hyalinization. Case 3 had a predominantly myxoid stroma with a rich network of thin-walled blood vessels. The tumor cells exhibited mild nuclear atypia with 1-3 mitotic figures per 50 high-power fields. All three cases showed diffuse and strong expression of CD10. Two of the three cases showed nuclear expression of TFE3, while one case showed diffuse and strong expression of desmin and S-100. Targeted RNA sequencing revealed PHF1 (ex12)::TFE3 (ex7) fusion in two cases and MEAF6 (ex5)::PHF1 (5′UTR) fusion in one case, which were further confirmed by FISH study. All three patients underwent tumor resection. Two showed no recurrence during follow-up periods of 98 months and 15 months, respectively, while one experienced local recurrence at 12 months postoperatively.Conclusions:OFMT with rare fusion subtypes often exhibits atypical histological and immunophenotypic features, and lacks a characteristic bony shell. Incorporating TFE3 into the diagnostic IHC panel greatly aids in screening for the cases with rare PHF1::TFE3 fusions. Familiarity with the histological and immunophenotypic characteristics, and differential diagnostic points of these rare OFMT subtypes, is essential for judicious use of molecular genetic tools in achieving a definitive diagnosis.

Objective To explore the mechanism of electroacupuncture in inhibiting peripheral sensitization of visceral pain in mice with irritable bowel syndrome(IBS)based on TRPV1/Ras/p38MAPK signaling pathway.Methods Totally 28 male SPF-grade C57BL/6 mice were randomly divided into normal group,model group,electroacupuncture group and inhibitor group.The IBS visceral hypersensitivity model was induced by trinitrobenzene sulfonic acid enema.Four weeks post-modeling,electroacupuncture group and inhibitor group were given electroacupuncture at bilateral"Zusanli"and intraperitoneal injection of the TRPV1 receptor inhibitor respectively for consecutive 7 d.Abdominal withdrawal reflex(AWR)score was used to evaluate visceral pain in mice,HE staining was used to observe the morphology of colonic tissue,Western blot was used to detect the protein expressions of TRPV1,p-p38,tumor necrosis factor(TNF)-α and interleukin(IL)-6 in colonic tissue,ELISA was used to detect the Ras-GTP content in colonic tissue,and RT-qPCR was used to detect the mRNA expression of TRPV1 and p38 in colonic tissue.Results Compared with the normal group,AWR score under different pressure were significantly increased in the model group(P<0.05,P<0.01),the expression of TRPV1,p-p38,TNF-α and IL-6 protein in colonic tissue significantly increased(P<0.05,P<0.01,P<0.001),the content of Ras-GTP significantly increased(P<0.001),the expression of TRPV1 and p38 mRNA significantly increased(P<0.001).Compared with the model group,AWR score under different pressure were significantly decreased in electroacupuncture group and inhibitor group(P<0.05,P<0.01),the expression of TRPV1,p-p38,TNF-α and IL-6 protein in colonic tissue significantly decreased(P<0.05,P<0.01,P<0.001),the content of Ras-GTP significantly decreased(P<0.01,P<0.001),and the expressions of TRPV1 and p38 mRNA significantly decreased(P<0.05,P<0.01,P<0.001).There was no significant change in the morphology of colonic tissue in each group of mice.Conclusion Electroacupuncture can effectively alleviate visceral hypersensitivity in IBS mice,and its analgesic effect may be related to the inhibition of the TRPV1/Ras/p38MAPK signaling pathway in colonic tissue.

Objective To analyze the effects of activating transcription factor 3(ATF3)pathway mediated by circSLC8Al on oxidative stress and iron activity of epileptic cells.Methods An epileptic cell model was established using human neuronal-hippocampal cells through Mg2+-free method.The expression levels of circSLC8A1 and ATF3 in healthy control group and model group were detected.Plasmid transfection was used to establish circSLC8A1 knockout group,ATF3 knockout group,circSLC8A1 knockout+ATF3 overexpression group,and ATF3 knockout+circSLC8A1 overexpression group.After 6h transfection,cells were cultured in normal medium for 48h.The cell viability,iron activity,reactive oxygen species(ROS),lactate dehydrogenase(LDH)and glutathione(GSH)of the different intervention groups were detected and compared.Results The expression levels of circSLC8A1,ATF3,ROS,LDH and iron activity in model group were significantly higher than those in healthy control group,while cell activity and GSH expression were significantly lower than those in healthy control group(P＜0.05).Knocking out circSLC8A1 can significantly reduce the expression of circSLC8A1 in epileptic model cells,while knocking out ATF3 can significantly reduce the expression of ATF3 in epileptic model cells(P＜0.05).Knocking out circSLC8A1 or ATF3 will increase the cell viability,decrease the iron activity and relieve the oxidative stress in epileptic model cells.Knocking out circSLC8A1 and overexpressing ATF3 can reverse the above trend,but knocking out ATF3 and overexpressing circSLC8A1 will not lead to the above phenomenon.Conclusion circSLC8A1 can influence the cell activity,oxidative stress and iron activity process of epileptic model cells by mediating ATF3 pathway,which provides some reference for the mechanism of epilepsy and its targeted therapy.

Objective To explore the influencing factors of delayed bleeding after endoscopic gastric polypectomy in antithrombotic patients.Methods A retrospective case-control study was conducted,42 patients who experienced delayed bleeding after endoscopic gastric polypectomy during antithrombotic therapy at Dongguan Eighth People's Hospital from April 2021 to April 2024 were selected as the bleeding group,and 84 patients who underwent the same surgery during the same period without delayed postoperative bleeding were selected as the non-bleeding group.The general data,polyps and antithrombotic treatment of the two groups were compared.The influencing factors of delayed bleeding after endoscopic gastric polypectomy in patients with antithrombotic treatment were analyzed by conditional Logistic regression,and the predictive value was evaluated by the receiver operating characteristic(ROC)curve.Results The maximum polyp diameter of the bleeding group was larger than that of the non-bleeding group(P<0.05).There were significant differences in the number of antithrombotic drugs and the management of antithrombotic drugs between the two groups(P<0.05).Logistic regression analysis showed that the large maximum polyp diameter before surgery(OR=4.056,95%CI:2.116 to 7.775),the use of multiple antithrombotic drugs before surgery(OR=5.308,95%CI:1.516 to 18.583),and the withdrawal of all antithrombotic drugs but the application of renal replacement therapy(OR=7.167,95%CI:1.921 to 26.734)were the risk factors for postoperative delayed bleeding(P<0.05).ROC curve showed the area under the curve of the large maximum polyp diameter before surgery,the use of multiple antithrombotic drugs before surgery,and the withdrawal of all antithrombotic drugs but the application of renal replacement therapy for predicting postoperative delayed bleeding were 0.882(95%CI:0.812 to 0.932),0.702(95%CI:0.614 to 0.781)and 0.746(95%CI:0.661 to 0.820).Conclusion The large maximum polyp diameter before surgery,preoperative use of multiple antithrombotic drugs,and the application of renal replacement therapy are influencing factors for delayed bleeding after endoscopic gastric polypectomy in antithrombotic patients.

Objective:To evaluate the long-term effectiveness of lingual mucosal graft ureteroplasty (LMGU) for managing long-segment (≥5 cm) ureteral strictures in a multi-institutional cohort of patients.Methods:A multi-center retrospective case series study was conducted on clinical data from 42 patients undergoing LMGU for long-segment ureteral strictures (≥5 cm) across five institutions between February 2017 and June 2024. The cohort comprised 31 males and 11 females, with an age of (43.4±12.0) years (range: 15 to 64 years) and a body mass index of (24.6±2.6) kg/m2 (range: 16.0 to 30.0 kg/m2). Strictures involved the left ureter in 24 cases and right ureter in 18 cases, demonstrating a stricture length of (6.4±1.5) cm (range: 5.0 to 11.5 cm). Surgical interventions included either onlay ureteroplasty or augmented anastomotic ureteroplasty, selected according to intraoperative findings. Intraoperative parameters, postoperative complications, and follow-up outcomes were analyzed.Results:Laparoscopic surgery was performed in 22 cases and robot-assisted surgery in 20 cases. Among the 42 patients, 22 underwent onlay ureteroplasty while 20 received augmented anastomotic ureteroplasty. The graft length was (5.9±1.8) cm (range: 3.0 to 12.0 cm), operative time (191.5±55.6) minutes (range: 105.0 to 350.0 minutes), and intraoperative estimated blood loss (86.7±73.6) ml (range: 10.0 to 400.0 ml). All procedures were successfully completed without conversion to open surgery. The postoperative hospital stay was (7.6±2.0) days (range: 4.0 to 15.0 days), with double-J stent removal at 6 to 8 weeks postoperatively. During a follow-up of (49.1±25.0) months (range: 12.0 to 99.0 months), no stricture recurrence was observed in any patient.Conclusion:LMGU is a safe, feasible, and effective long-term technique for managing long-segment (≥5 cm) ureteral strictures.

This study was aimed at investigating the effects of demethylase fat mass and obesity-associated protein(FTO)and methyltransferase methyltransferase like protein 3(METTL3),key molecules in N6-methyladenosine(m6A)modification,on the replication and proliferation of Japanese encephalitis virus(JEV).Recombinant lentiviruses were generated by packaging the FTO and green fluorescent protein into lentiviral vectors.Neuro2a cells,a mouse neuroblastoma cell line,were infected with the lentivirus,and stable FTO-expressing cell lines were obtained through puromycin selection.Successful overexpression of FTO was confirmed through fluorescence microscopy,real-time quantitative PCR,and western blot analysis.When Neuro2a cells overexpressing FTO were infected with JEV,the overexpression of FTO decreased JEV replication in the cells,and increased the expression of interferon(IFN)and related molecules.Additionally,treatment of JEV-infected Neuro2a cells with the METTL3-specific inhibitor STM2457 resulted in a dose-dependent decrease in JEV replication and viral protein expression.These findings suggested that lowering m6A methylation levels inhibits JEV replication,thus shedding light on the regulatory role of methylation modification in JEV replication.