Purpose This study aimed to investigate the expression,function,and molecular mechanisms of ZNF384 in esophageal squamous cell carcinoma(ESCC),as well as its role in tumor progression and chemoresistance.Methods The expression of ZNF384 in ESCC cell lines and tissues was assessed using RT-qPCR.Correlations with TNM stage,invasion depth,lymph node metastasis,and prognosis were evaluated.In vitro assays were performed to examine the effects of ZNF384 on ESCC cell proliferation,migration,invasion,and chemosensitivity.Dual-luciferase reporter assays were conducted to determine the interaction between ZNF384 and FZD3,and to assess the activation of the Wnt signaling pathway.Results ZNF384 expression was significantly upregulated in ESCC cell lines and tissues(P＜0.01).Elevated ZNF384 expression was associated with advanced TNM stage,greater invasion depth,lymph node metastasis,and poor prognosis(P＜0.05).Functional assays demonstrated that ZNF384 overexpression promo-ted ESCC cell proliferation,migration,and invasion(all P＜0.01),whereas ZNF384 knockdown inhibited these processes and enhanced chemosensitivity to cisplatin(all P＜0.01).Mechanistic studies showed that ZNF384 directly bound to the FZD3 promoter,upregulated FZD3 expression,and activated the Wnt signaling pathway(P＜0.05).Overexpression of FZD3 partially reversed the inhibitory effects of ZNF384 knockdown on cell malignancy and chemore-sistance(P＜0.05).Conclusion ZNF384 promotes ESCC progression and reduces chemosensitivity through activa-tion of the FZD3/Wnt signaling pathway,suggesting its potential as a therapeutic target in ESCC.

Idiopathic Pulmonary Fibrosis(IPF)is a progressive disease characterized by declining respiratory function and high mortality.Macrophages play a pivotal role in its pathogenesis.Through polarization into pro-inflammatory M1 and pro-fibrotic M2 phenotypes,they contribute to a complex immunoregulatory network.In the early disease stages,M1 macrophage-mediated inflammation causes lung tissue injury,while M2 macrophages drive fibrogenesis by releasing pro-fibrotic factors such as TGF-β.Recent research has revealed that exosomes derived from macrophages serve as carriers for miRNAs,with specific miRNAs(e.g.,miR-328,miR-142-3p)demonstrating significant roles in pulmonary fibrosis.miR-328 promotes fibroblast proliferation and accelerates collagen deposition,whereas miR-142-3p attenuates fibrosis by modulating the TGF-β signaling pathway.Targeted intervention against these macrophage-associated miRNAs shows potential for clinical translation,potentially offering novel approaches for the early diagnosis and targeted therapy of IPF.However,translating these strategies into clinical practice requires overcoming challenges related to production and delivery systems.In conclusion,a deeper understanding of the mechanisms and translational applications of macrophage-derived miRNAs in IPF holds promise for ultimately improving patient prognosis and clinical outcomes.

Idiopathic Pulmonary Fibrosis(IPF)is a progressive disease characterized by declining respiratory function and high mortality.Macrophages play a pivotal role in its pathogenesis.Through polarization into pro-inflammatory M1 and pro-fibrotic M2 phenotypes,they contribute to a complex immunoregulatory network.In the early disease stages,M1 macrophage-mediated inflammation causes lung tissue injury,while M2 macrophages drive fibrogenesis by releasing pro-fibrotic factors such as TGF-β.Recent research has revealed that exosomes derived from macrophages serve as carriers for miRNAs,with specific miRNAs(e.g.,miR-328,miR-142-3p)demonstrating significant roles in pulmonary fibrosis.miR-328 promotes fibroblast proliferation and accelerates collagen deposition,whereas miR-142-3p attenuates fibrosis by modulating the TGF-β signaling pathway.Targeted intervention against these macrophage-associated miRNAs shows potential for clinical translation,potentially offering novel approaches for the early diagnosis and targeted therapy of IPF.However,translating these strategies into clinical practice requires overcoming challenges related to production and delivery systems.In conclusion,a deeper understanding of the mechanisms and translational applications of macrophage-derived miRNAs in IPF holds promise for ultimately improving patient prognosis and clinical outcomes.

Purpose This study aimed to investigate the expression,function,and molecular mechanisms of ZNF384 in esophageal squamous cell carcinoma(ESCC),as well as its role in tumor progression and chemoresistance.Methods The expression of ZNF384 in ESCC cell lines and tissues was assessed using RT-qPCR.Correlations with TNM stage,invasion depth,lymph node metastasis,and prognosis were evaluated.In vitro assays were performed to examine the effects of ZNF384 on ESCC cell proliferation,migration,invasion,and chemosensitivity.Dual-luciferase reporter assays were conducted to determine the interaction between ZNF384 and FZD3,and to assess the activation of the Wnt signaling pathway.Results ZNF384 expression was significantly upregulated in ESCC cell lines and tissues(P＜0.01).Elevated ZNF384 expression was associated with advanced TNM stage,greater invasion depth,lymph node metastasis,and poor prognosis(P＜0.05).Functional assays demonstrated that ZNF384 overexpression promo-ted ESCC cell proliferation,migration,and invasion(all P＜0.01),whereas ZNF384 knockdown inhibited these processes and enhanced chemosensitivity to cisplatin(all P＜0.01).Mechanistic studies showed that ZNF384 directly bound to the FZD3 promoter,upregulated FZD3 expression,and activated the Wnt signaling pathway(P＜0.05).Overexpression of FZD3 partially reversed the inhibitory effects of ZNF384 knockdown on cell malignancy and chemore-sistance(P＜0.05).Conclusion ZNF384 promotes ESCC progression and reduces chemosensitivity through activa-tion of the FZD3/Wnt signaling pathway,suggesting its potential as a therapeutic target in ESCC.

Objective:To construct a recombinant bioluminescent bacteriophage (HT7) targeting Escherichia coli, and evaluate its ability to identify Escherichia coli. Methods:Initially, pCRISPR-sg (1-10) and PFN-1000 plasmid strains were constructed by genetic engineering, and the most efficient small guild RNA (sgRNA) were screened by bilayer plate. By the gene editing technique, which comprised homologous recombination and clustered regularly interspaced short palin dromic repeats (CRISPR)-Cas system, the Nanoluc luciferase gene was integrated into the downstream non-coding region of 10A gene of T7 phage, to constructe the bioluminescent phage HT7 successfully. The difference of biological characteristics between HT7 phage and T7 phage was evaluated by plaque assay and liquid amplification assay. In addition, 51 strains of Escherichia coli, 20 strains of Klebsiella pneumoniae, 14 strains of Staphylococcus aureus, 6 strains of Enterococcus faecium, 5 strains of Enterococcus faecalis, 3 strains of Acinetobacter baumannii and 1 strain of Pseudomonas aeruginosa were collected and isolated to evaluate the limit of detection and specificity of HT7 phage. Results:Among the 10 CRISPR-targeted cleavage systems constructed, sgRNA8 exhibited the highest cleavage efficiency, with a cleavage rate of 0.18. After three rounds of recombination screening using the pCas9/pCRISPR/PFN-1000 triple-plasmid system, PCR validation yielded recombinant phage bands at 2 798 bp, indicating the successful construction of the HT7 phage. The recombinant phage showed significant differences in biological characteristics in terms of lysis efficiency ( P<0.001), one-step growth curve ( P=0.001), and infection multiplicity ( P=0.031). Both lysis burst time and log growth node were extended by 10 min, with the optimal infection multiplicity being 0.1. Clinical sample testing identified lysis of 6 strains of Escherichia coli within 4.5 h, while other strains remained unaffected, with detection of pathogenic bacteria below 10 CFU/ml. Conclusions:The developed pCas9/pCRISPR/PFN-1000 triple-plasmid editing system efficiently edits the bacteriophage genome. The constructed HT7 fluorescent bacteriophage enables the detection of Escherichia coli below 10 CFU/ml within 4.5 hours, demonstrating low detection limits and high detection specificity.

Objective:This study aimed to explore the status of radiological Kashin-Beck disease (KBD) among school-aged children in Chamdo City, Tibet, through a 3-year monitoring survey, providing epidemiological evidence for prevention and control strategies.Methods:The target areas for this study were Luolong, Bianba, and Basu counties in Chamdo City, Tibet Autonomous Region, identified as having the most severe historical cases of KBD. Children aged 7-12 years attending school were enrolled as study subjects. Anteroposterior X-ray films of the right-hand were taken, and radiological diagnoses were made based on the "Diagnosis of Kashin-Beck Disease" criteria (WS/T 207-2010). Two experienced researchers independently reviewed the X-rays, and intra- and inter-group consistency were assessed using weighted Kappa values and percentage agreement. Cross-sectional surveys were conducted in 2017 and 2020 to describe the X-ray detection rates of KBD, and logistic regression analysis was employed to construct a predictive model of risk factors for radiological KBD cases.Results:In 2017, a total of 5,711 children aged 7-12 years in Chamdo City, Tibet, participated in the baseline cross-sectional survey (average age 9.2 years, 48.0% female), with 28 cases of radiological KBD. The age- and gender-standardized prevalence rate was 0.527%. In 2020, 6,771 participants (average age 9.3 years, 49.5% female) underwent a second cross-sectional survey, with 9 cases of radiological KBD and a standardized prevalence rate of 0.134%. Logistic regression analysis indicated that older age [ OR=2.439, 95% CI(1.299, 4.580), P=0.006] and female gender [ OR=8.157, 95% CI(1.016, 65.528), P=0.048] were independent risk factors for radiological KBD cases. Conversely, higher residential altitude, under the premise of Tibet's high altitude, was a protective factor [ OR=0.995, 95% CI(0.990, 0.999), P=0.032). Conclusion:The radiographically positive detection rate of KBD among school-aged children in Chamdo City, Tibet Autonomous Region, is at an extremely low level and showing a declining trend, reaching the historical standard in 2020. Considering the absence of positive signs in affected children, it suggests that local KBD has been effectively eliminated.

In recent years, endovascular therapy has become the most important progress in the field of the treatment of acute ischemic stroke caused by large vessel occlusion. However, the vascular recanalization shown by imaging after endovascular treatment cannot fully translate into effective tissue reperfusion and functional outcome, a phenomenon known as "futile recanalization". Combined neuroprotective therapy after vascular recanalization is expected to reduce the occurrence of futile recanalization and improve the outcome of patients. This article briefly summarizes the main application progress of commonly used neuroprotective therapies in clinical practice (edaravone dexborneol, glucocorticoids, hypothermia, and remote ischemic conditioning). It explores the trend and direction of combining endovascular therapy and neuroprotective therapy for patients with acute ischemic stroke caused by large vessel occlusion, and provides further reference and suggestions for intervention measures after endovascular therapy.

Extracellular matrix hydrogel has been widely used in three-dimensional cell culture, organoid construction, tissue defect repair, biological three-dimensional printing and tissue engineering due to its excellent bioactivity, biocompatibility, bioinducibility, biodegradability and minimally invasive injection. Adipose tissue has the advantages of sufficient sources, easy access, and rich content of extracellular matrix components. It has become a research hotspot in the field of tissue repair and regenerative medicine in recent years. This article reviews its preparation, detection and application, in order to provide a reference for relevant researchers.

As a group of nonspecific inflammatory diseases affecting the intestine, inflammatory bowel disease (IBD) exhibits the characteristics of chronic recurring inflammation, and was proven to be increasing in incidence (Kaplan, 2015). IBD induced by genetic background, environmental changes, immune functions, microbial composition, and toxin exposures (Sasson et al., 2021) primarily includes ulcerative colitis (UC) and Crohn's disease (CD) with complicated clinical symptoms featured by abdominal pain, diarrhea, and even blood in stools (Fan et al., 2021; Huang et al., 2021). UC is mainly limited to the rectum and the colon, while CD usually impacts the terminal ileum and colon in a discontinuous manner (Ordás et al., 2012; Panés and Rimola, 2017). In recent years, many studies have suggested the lack of effective measures in the diagnosis and treatment of IBD, prompting an urgent need for new strategies to understand the mechanisms of and offer promising therapies for IBD.
Chronic Disease ; Colitis, Ulcerative/therapy* ; Crohn Disease/epidemiology* ; Diarrhea ; Homeodomain Proteins ; Humans ; Inflammatory Bowel Diseases ; Mesenchymal Stem Cells/cytology* ; MicroRNAs ; RNA, Long Noncoding ; Recurrence ; Umbilical Cord/cytology*

The family of voltage-gated potassium Kv2 channels consists of the Kv2.1 and Kv2.2 subtypes. Kv2.1 is constitutively highly phosphorylated in neurons and its function relies on its phosphorylation state. Whether the function of Kv2.2 is also dependent on its phosphorylation state remains unknown. Here, we investigated whether Kv2.2 channels can be phosphorylated by protein kinase C (PKC) and examined the effects of PKC-induced phosphorylation on their activity and function. Activation of PKC inhibited Kv2.2 currents and altered their steady-state activation in HEK293 cells. Point mutations and specific antibodies against phosphorylated S481 or S488 demonstrated the importance of these residues for the PKC-dependent modulation of Kv2.2. In layer II pyramidal neurons in cortical slices, activation of PKC similarly regulated native Kv2.2 channels and simultaneously reduced the frequency of action potentials. In conclusion, this study provides the first evidence to our knowledge that PKC-induced phosphorylation of the Kv2.2 channel controls the excitability of cortical pyramidal neurons.
Action Potentials ; HEK293 Cells ; Humans ; Protein Kinase C/metabolism* ; Pyramidal Cells/enzymology* ; Shab Potassium Channels/genetics*