BACKGROUND:U937 cells can be used as a cell model for studying the biological characteristics,signaling pathways,and therapeutic targets of acute myeloid leukemia.Although it has been reported that long non-coding RNA KIAA0125 is highly expressed in acute myeloid leukemia,its biological function in U937 cells remains unclear,and its mechanism of action in the occurrence and development of acute myeloid leukemia needs to be further clarified. OBJECTIVE:To investigate the expression level of long non-coding RNA KIAA0125 in peripheral blood of patients with acute myeloid leukemia and its effect on the proliferation and apoptosis of U937 cells. METHODS:RNA-sequencing was used to analyze the bone marrow monocyte samples from acute myeloid leukemia patients,and the differentially expressed gene long non-coding RNA KIAA0125 was screened.The expression of long non-coding RNA KIAA0125 in peripheral blood of patients with acute myeloid leukemia was detected by qRT-PCR.The relationship between long non-coding RNA KIAA0125 mRNA expression and prognosis in bone marrow cells of 173 acute myeloid leukemia patients and 70 healthy people was statistically analyzed by GEPIA database.Subsequently,recombinant lentivirus technology and CRISPR/Cas9-SAM technology were used to construct U937 cell lines with knockdown/overexpression of long non-coding RNA KIAA0125.qRT-PCR was used to detect the knockdown/overexpression efficiency of long non-coding RNA KIAA0125.Next,CCK-8 assay,flow cytometry,and western blot assay were used to detect the effects of knockdown/overexpression of long non-coding RNA KIAA0125 on the proliferation and apoptosis of U937 cells.Finally,western blot assay was used to detect the effect of knockdown/overexpressed long non-coding RNA KIAA0125 on Wnt/β-catenin signaling pathway-related proteins. RESULTS AND CONCLUSION:(1)The results of qRT-PCR showed that long non-coding RNA KIAA0125 was highly expressed in peripheral blood of acute myeloid leukemia patients.The results of GEPIA database showed that long non-coding RNA KIAA0125 was highly expressed in bone marrow cells of acute myeloid leukemia patients,and the high expression group had worse overall survival.(2)The knockdown efficiency of long non-coding RNA KIAA0125 in knockdown group was 70%,and the U937 cells that stably down-regulated long non-coding RNA KIAA0125 expression were successfully constructed.The expression of long non-coding RNA KIAA0125 in overexpression group was four times that of vector group,and stable U937 cells were successfully constructed.(3)Knockdown of long non-coding RNA KIAA0125 inhibited the proliferation of U937 cells and promoted their apoptosis.Overexpression of long non-coding RNA KIAA0125 promoted the proliferation of U937 cells but had no significant effect on the apoptosis of U937 cells.(4)Knockdown of long non-coding RNA KIAA0125 inhibited the activity of Wnt/β-catenin signaling pathway,while overexpression of long non-coding RNA KIAA0125 activated Wnt/β-catenin signaling pathway.These results confirm that long non-coding RNA KIAA0125 is highly expressed in acute myeloid leukemia peripheral blood.Long non-coding RNA KIAA0125 may affect the proliferation and apoptosis of U937 cells by regulating the Wnt/β-catenin signaling pathway,and may be a potential prognostic marker for acute myeloid leukemia.

Objective To analyze the proportion and clinicopathological characteristics of HER2-positive breast cancer patients with BRCA1/2 pathogenic variants, and their response to neoadjuvant anti-HER2 targeted therapy. Methods The clinicopathological data of 531 breast cancer patients with germline BRCA1/2 pathogenic variants (201 with BRCA1 variants and 330 with BRCA2 variants) were analyzed. Results Among the 201 BRCA1 and 330 BRCA2 variants, 17 (8.5%) and 42 (12.7%) HER2-positive breast cancer cases were identified, respectively, accounting for 11.1% of all BRCA1/2-mutated breast cancers. Compared with BRCA1/2-mutated HR-positive/HER2-negative patients, HER2-positive patients did not present any significant differences in clinicopathological features; however, compared with triple-negative breast cancer patients, HER2-positive patients had a later onset age and lower tumor grade. Among the 17 patients who received neoadjuvant anti-HER2 targeted therapy, 10 cases achieved pCR (58.8%), whereas 7 cases did not (41.2%). Conclusion HER2-positive breast cancer accounts for more than 10% of BRCA1/2-mutated patients. Approximately 40% of these patients fail to achieve pCR after neoadjuvant targeted therapy. This phenomenon highlights the possibility of combining anti-HER2 targeted agents with poly (adenosine diphosphate-ribose) polymerase inhibitors.

ObjectiveTo investigate the effect and mechanism of Bukan Yilidan on perimenopausal psycho-cardiac disease by mitochondrial autophagy mediated by dynamin-related protein 1（Drp1）/phosphatase and tensin homolog（PTEN） induced putative kinase 1（PINK1）/Parkin pathway. MethodsSixty rats were randomly divided into the blank group， model group， western medicine group（isosorbide mononitrate 7.2 mg·kg^-1+sertraline hydrochloride tablets 18 mg·kg^-1）， Bukan Yilidan low， medium and high dose groups（2.59， 5.18， 10.35 g·kg^-1）， with 10 rats in each group. Except for the blank group， the rat model of perimenopausal psycho-cardiac disease was prepared by ovariectomy（OVX） combined with high-fat feeding， chronic unpredictable mild stress（CUMS） and subcutaneous multi-point injection of isoproterenol hydrochloride. After successful modeling， the general state and tongue image of rats were observed. The depression status of rats in vivo was evaluated by open field test， sucrose preference test， forced swimming immobility time and grip strength value， and the cardiac function of rats was evaluated by electrocardiogram and echocardiography. The levels of serum norepinephrine（NE）， dopamine（DA） and 5-hydroxytryptamine（5-HT） were detected by enzyme-linked immunosorbent assay（ELISA）， and biochemical detection was used to assess myocardial injury by measuring serum levels of high-density lipoprotein cholesterol（HDL-C）， low-density lipoprotein cholesterol（LDL-C）， triglyceride（TG）， total cholesterol（TC）， aspartate aminotransferase（AST）， alanine aminotransferase（ALT）， lactate dehydrogenase（LDH） and creatine kinase isoenzyme（CK-MB）. Hematoxylin-eosin（HE） and Nissl staining were used to observe the pathological status of hippocampus and myocardial tissue in rats， the status of mitophagosomes in hippocampus and myocardium was observed by transmission electron microscope（TEM）， and Western blot was used to detect the contents of Drp1， mitochondrial fusion protein 2（Mfn2）， optic atrophy protein 1（OPA1）， PINK1， Parkin， p62 and microtubule-associated protein light chain 3B（LC3B） in hippocampus and myocardium. ResultsCompared with the blank group， the food intake and water intake of rats in the model group decreased， the hair was dark yellow， the gloss and smoothness decreased， the spirit was depressed， the tongue was light purple or dark purple， accompanied by petechiae or ecchymosis， the sublingual collaterals were purple and black， and the tongue coating was white and smooth. The indexes of open field test， grip strength and sucrose preference of rats decreased significantly， and the immobility time of forced swimming increased significantly（P<0.01）. Electrocardiogram and echocardiography showed that ST segment was significantly depressed， and left ventricular fractional shortening（LVFS） and left ventricular ejection fraction（LVEF） were significantly decreased（P<0.05， P<0.01）. Pathological observation showed that the number of hippocampal neurons and myocardial cells decreased， and the structural damage was obvious. The levels of serum TC， TG， LDL-C， CK-MB， LDH， AST and ALT increased， while the levels of HDL-C， 5-HT， DA and NE decreased（P<0.05， P<0.01）. TEM showed obvious mitochondrial damage in hippocampus and myocardial tissue. The protein expressions of Drp1， PINK1， Parkin and p62 in hippocampus and myocardium were increased， while the protein expressions of OPA1， Mfn2 and LC3BⅡ/Ⅰ were decreased（P<0.05， P<0.01）. Compared with the model group， the mental state， body curling up， fear of cold and other symptoms of rats in each administration group were improved， and the degree of pale purple or dark purple tongue was reduced. The scores of open field test， grip strength， sucrose preference， LVFS and LVEF were increased， and the immobility time of forced swimming was shortened（P<0.05， P<0.01）. The ST segment of electrocardiogram had a significant recovery（P<0.01）， pathological observation showed that the damage of nerve cells and myocardial tissue was improved. The levels of serum TC， TG， LDL-C， CK-MB， LDH， AST and ALT decreased， while the levels of HDL-C， 5-HT， DA and NE increased（P<0.05， P<0.01）. TEM showed that mitochondrial damage was reduced in hippocampal neurons and cardiomyocytes with visible mitochondrial autophagosomes. The protein expressions of Drp1， PINK1， Parkin and p62 in hippocampus and myocardium were decreased， while the protein expressions of OPA1， Mfn2 and LC3BⅡ/Ⅰ were increased（P<0.05， P<0.01）. ConclusionBukan Yilidan can alleviate depression， lipid metabolism disorder and myocardial ischemia injury in rats with perimenopausal psycho-cardiac disease， and its mechanism may be related to inhibiting Drp1/PINK1/Parkin signaling pathway and enhancing mitochondrial autophagy.

ObjectiveTo investigate the awareness and clinical practice of guidelines for the prevention and treatment of chronic hepatitis B （2022 edition） among clinicians. MethodsFrom July 19 to December 31， 2024， a self-designed electronic questionnaire was distributed via the WeChat mini program to collect related data from 1 588 clinicians nationwide， including their awareness and practice based on 18 questions regarding testing and referral， diagnosis and treatment， and follow-up. ResultsAmong all respondents， only 350 clinicians correctly understood all the updated key points of antiviral indications and treatment for special populations in the 2022 edition of guidelines for the prevention and treatment of chronic hepatitis B， with an overall awareness rate of 22.0%. Only 20% ‍—‍ ‍40% of the patients with positive HBV DNA and an age of >30 years receive antiviral therapy， while 80% ‍—‍ ‍100% of the patients with positive HBV DNA and a family history of hepatitis B cirrhosis or hepatocellular carcinoma receive antiviral therapy. The median follow-up rates at 1 year， 3 years， and 5 years were 67.5% 57.5% and 47.5%，respectively， showing a trend of gradual reduction， which might be associated with the influencing factors such as insufficient time for follow-up management by clinicians， insufficient awareness of the disease among patients， and poor adherence to follow-up. ConclusionThere is a gap between the awareness and practice of guidelines for the prevention and treatment of chronic hepatitis B （2022 edition） among clinicians. It is recommended to further strengthen training and focus on the whole process of “detection， diagnosis， treatment， and management” for patients with chronic hepatitis B in healthcare institutions， in order to promote the implementation of the guidelines.

Objective To investigate the role and underlying mechanism of C1q-like protein 4 (C1ql4) in regulating the characteristics of breast cancer stem cells. Methods qRT-PCR was used to detect the expression of C1ql4 in breast cancer and normal breast epithelial cell lines, as well as to verify the transfection efficiency of C1ql4. Western blot analysis was employed to examine the phosphorylation levels of AKT, IKK, and IκB in different groups. An AKT activator was added to MDA-MB-231 cells with C1ql4 knockdown, whereas inhibitors targeting AKT, IKK, IκB, and NF-κB nuclear translocation were separately introduced to C1ql4-overexpressing MCF-7 cells. The nuclear translocation of NF-κB, expression levels of the target genes TNF-α and IL-1β, formation ability of tumorspheres, and proportion of CD44⁺/CD24^−/low stem-like subgroups were analyzed. Results C1ql4 expression in breast cancer cell lines was significantly upregulated compared with that in normal breast epithelial cells. Western blot analysis showed that p-AKT/AKT, p-IKK/IKK, and p-IκB/IκB ratios markedly reduced in C1ql4-knockdown MDA-MB-231 cells (all P<0.05) but significantly increased in C1ql4-overexpressing MCF-7 cells (all P<0.05). Rescue experiments demonstrated that the addition of an AKT activator to C1ql4-knockdown MDA-MB-231 cells resulted in the enhanced nuclear translocation of NF-κB, the increased nuclear/cytoplasmic NF-κB ratios, the elevated TNF-α and IL-1β expression levels, and significant recovery of tumorsphere formation ability and the proportion of CD44⁺/CD24^−/low stem-like subpopulations (all P<0.05). Conversely, in C1ql4-overexpressing MCF-7 cells, treatment with AKT, IKK, IκB, or NF-κB nuclear translocation inhibitors led to a reduction in NF-κB nuclear translocation, decreased nuclear/cytoplasmic NF-κB ratios, and declines in TNF-α and IL-1β expression levels, tumorsphere formation ability, and the CD44⁺/CD24^−/low subpopulation (all P<0.05). Conclusion C1ql4 promotes the translocation of NF-κB from the cytoplasm to the nucleus through the PI3K/AKT/NF-κB signaling pathway and enhances the expression of stemness in breast cancer cells.

Objective: To analyze the prevalence and determinants of comorbid overweight/obesity and elevated blood pressure among primary and secondary school students in Yangzhou City, and to explore the association between sleep patterns, eating behavior and the comorbidity of overweight/obesity and elevated blood pressure, so as to provide reference for developing prevention strategies targeting common comorbidities in students. Methods: By using stratified cluster random sampling, a total of 8 735 primary and secondary school students were selected from 36 schools in six counties of Yangzhou from October to November 2023. Students underwent physical examinations and a questionnaire survey was conducted using the questionnaire on students health status and influencing factors. The Chi square test was used to compare the detection rate of comorbid overweight/obesity and elevated blood pressure in different groups of primary and secondary school students. The Logistic regression model was used to explore the association between sleep and dietary behaviors and their combined effects and coexistence. Results: The detection rate of comorbid overweight/obesity and elevated blood pressure among primary and secondary school students in Yangzhou was 9.85%, which was higher among boys (12.14%) than girls (7.59%)( χ 2=50.86, P <0.01). After controlling for gender, residence, educational stage, parental education, smoking, drinking, and moderate to vigorous exercise, multivariate Logistic regression analysis showed that irregular breakfast consumption and inadequate daily sleep were associated with a higher risk of comorbidities compared with regular breakfast consumption and adequate daily sleep among overall and primary school students (overall: OR =1.52, 95% CI =1.18- 1.96 , primary school students: OR =2.79, 95% CI =1.61-4.82)(both P <0.05). From the perspective of primary school students of different genders, the risk of comorbidities in girls who consumed breakfast irregularly and had inadequate daily sleep was 3.59 times higher than that in girls who consumed breakfast irregularly and had inadequate daily sleep (95% CI =1.65-7.82, P <0.01). Conclusion The sleep patterns and breakfast behaviors of primary and secondary school students are found to be associated with comorbid overweight/obesity and elevated blood pressure, especially in primary school girls.

OBJECTIVE To provide a reference for standardizing the conduct of positron-emitting radiopharmaceuticals’ phase 0 clinical trials （hereinafter referred to as “phase 0 clinical trials”） and advancing the development of innovative drug by medical institutions. METHODS Initiated by the First Affiliated Hospital of Jinan University， a panel of experts consisting of pharmacy， clinical medicine and medical ethics from multiple institutions was established to investigate the current landscape， and discuss the necessary conditions， procedures， and other aspects for conducting phase 0 clinical trials in medical institutions by integrating relevant national policies， regulations and expert consensus. Finally， an agreement was reached to formulate this consensus. RESULTS & CONCLUSIONS Currently， most medical institutions have deficiencies in pharmaceutical care during the management of radiopharmaceuticals and the phase 0 clinical trials. In conjunction with the Expert Consensus on the Establishment of Nuclear Pharmacist Positions， this consensus explicitly defines the responsibilities of nuclear pharmacists in the phase 0 clinical trials on the basis of the Expert Consensus for the Application of Positron Emission Tomography Radioligands for Translational Study in the Phase 0 Clinical Trials （2020 edition）， providing a guidance for high-quality participation of nuclear pharmacists from medical institutions in China in phase 0 clinical research. Additionally， in consideration of some constraints imposed by current relevant regulations， this consensus also proposes strategic recommendations， such as encouraging medical institutions to form a consortium， leading to the establishment of dedicated bases or industrial parks， holding significant implications to strengthen institutional capacity for advancing radiopharmaceutical innovation through phase 0 clinical trials.

In recent years, the deep integration of artificial intelligence (AI) into medical education has created new opportunities for teaching Biochemistry and Molecular Biology, while also offering innovative solutions to the pedagogical challenges associated with protein structure and function. Focusing on the case of anaplastic lymphoma kinase (ALK) gene mutations in non-small-cell lung cancer (NSCLC), this study integrates AI into case-based learning (CBL) to develop an AI-CBL hybrid teaching model. This model features an intelligent case-generation system that dynamically constructs ALK mutation scenarios using real-world clinical data, closely linking molecular biology concepts with clinical applications. It incorporates AI-powered protein structure prediction tools to accurately visualize the three-dimensional structures of both wild-type and mutant ALK proteins, dynamically simulating functional abnormalities resulting from conformational changes. Additionally, a virtual simulation platform replicates the ALK gene detection workflow, bridging theoretical knowledge with practical skills. As a result, a multidimensional teaching system is established—driven by clinical cases and integrating molecular structural analysis with experimental validation. Teaching outcomes indicate that the three-dimensional visualization, dynamic interactivity, and intelligent analytical capabilities provided by AI significantly enhance students’ understanding of molecular mechanisms, classroom engagement, and capacity for innovative research. This model establishes a coherent training pathway linking “fundamental theory-scientific research thinking-clinical practice”, offering an effective approach to addressing teaching challenges and advancing the intelligent transformation of medical education.

ObjectiveThis study aims to explore the neurotoxicity mechanism of Dictamni Cortex by integrating network toxicology and metabolomics techniques. MethodsThe neurotoxicity targets induced by Dictamni Cortex were screened by the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP）， Traditional Chinese Medicine Information Database （TCM-ID）， and Comparative Toxicogenomics Database （CTD）. The target predictions of the components were performed by the Swiss Target Prediction tool. Neurotoxicity-related targets were collected from the Pharmacophore Mapping and Potential Target Identification Platform （PharmMapper）， GeneCards Human Gene Database （GeneCards）， DisGeNET Disease Gene Network （DisGeNET）， and Online Mendelian Inheritance in Man （OMIM）， and the intersection targets were identified. Protein-protein interaction （PPI） analysis， Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway analysis， and Gene Ontology （GO） enrichment analysis were conducted. A "drug-compound-toxicity target-pathway" network was constructed via Cytoscape software to display the core regulatory network. Based on the prediction results， the neurotoxicity mechanism of Dictamni Cortex in mice was verified by using hematoxylin-eosin （HE） staining， Nissl staining， enzyme-linked immunosorbent assay （ELISA）， quantitative real-time fluorescence polymerase chain reaction （Real-time PCR）， and Western blot. The effects of Dictamni Cortex on the metabolic profile of mouse brain tissue were further explored by non-targeted metabolomics. ResultsNetwork toxicology screening identified 13 compounds and 175 targets in Dictamni Cortex that were related to neurotoxicity. PPI network analysis revealed that serine/threonine-protein kinase （Akt1） and tumor protein 53 （TP53） were the core targets. Additionally， GO/KEGG enrichment analysis indicated that Dictamni Cortex may regulate the phosphatidylinositol 3-kinase （PI3K）/Akt pathway and affect oxidative stress and cell apoptosis， thereby inducing neural damage. The "Dictamni Cortex-compound-toxicity target-pathway-neural damage" network showed that dictamnine， phellodendrine， and fraxinellone may be the toxic compounds. Animal experiments showed that compared with those in the blank group， the hippocampal neurons in the brain tissue of mice treated with Dictamni Cortex were damaged. The level of superoxide dismutase （SOD） and acetylcholine （ACh） in the brain tissue was significantly reduced， while the content of malondialdehyde （MDA） was significantly increased. The level of Akt1 and p-Akt1 mRNAs and proteins in the brain tissue was significantly decreased， while the level of TP53 was significantly increased. Non-targeted metabolomics results showed that Dictamni Cortex could disrupt the level of 40 metabolites in mouse brain tissue， thereby regulating the homeostasis of 13 metabolism pathways， including phenylalanine， glycerophospholipid， and retinol. Combined analysis revealed that Akt1， p-Akt1， and TP53 were significantly correlated with phenylalanine， glycerophospholipid， and retinol metabolites. This suggested that Dictamni Cortex induced neurotoxicity in mice by regulating Akt1， p-Akt1， and TP53 and further modulating the phenylalanine， glycerophospholipid， and retinol metabolism pathways. ConclusionDictamni Cortex can induce neurotoxicity in mice， and its potential mechanism may be closely related to the activation of oxidative stress， inhibition of the PI3K/Akt signaling pathway， and regulation of phenylalanine， glycerophospholipid， and retinol metabolism pathways.

ObjectiveThis study aims to investigate the effect of Dictamni Cortex on intestinal barrier damage in rats and its mechanism by untargeted metabolomics and targeted metabolomics for short-chain fatty acids （SCFAs）. MethodsRats were randomly divided into a control group， a high-dose group of Dictamni Cortex （8.1 g·kg^-1）， a medium-dose group （2.7 g·kg^-1）， and a low-dose group （0.9 g·kg^-1）. Except for the control group， the other groups were administered different doses of Dictamni Cortex by gavage for eight consecutive weeks. Hematoxylin-eosin （HE） staining was used to observe the pathological changes in the ileal tissue. Enzyme-linked immunosorbent assay （ELISA） was employed to detect the level of cytokines， including tumor necrosis factor-α （TNF-α）， interleukin-6 （IL-6）， and interleukin-1β （IL-1β）， in the ileal tissue of rats. Quantitative real-time fluorescence polymerase chain reaction （Real-time PCR） technology was used to detect the expression level of tight junction proteins， including zonula occludens-1 （ZO-1）， Occludin， and Claudin-1 mRNAs， in the ileal tissue of rats to preliminarily explore the effects of Dictamni Cortex on intestinal damage. The dose with the most significant toxic phenotype was selected to further reveal the effects of Dictamni Cortex on the metabolic profile of ileal tissue in rats by non-targeted metabolomics combined with targeted metabolomics for SCFAs. ResultsCompared with the control group， all doses of Dictamni Cortex induced varying degrees of pathological damage in the ileum， increased TNF-α （P<0.01）， IL-6 （P<0.01）， and IL-1β （P<0.01） levels in the ileal tissue， and decreased the expression level of ZO-1 （P<0.05， P<0.01）， Occludin （P<0.01）， and Claudin-1 （P<0.05） in the ileal tissue， with the high-dose group showing the most significant toxic phenotypes. The damage mechanisms of the high-dose group of Dictamni Cortex on the ileal tissue were further explored by integrating non-targeted metabolomics and targeted metabolomics for SCFAs. The non-targeted metabolomics results showed that 21 differential metabolites were identified in the control group and the high-dose group. Compared with that in the control group， after Dictamni Cortex intervention， the level of 14 metabolites was significantly increased （P<0.05， P<0.01）， and the level of seven metabolites was significantly decreased （P<0.05， P<0.01） in the ileal contents. These metabolites collectively acted on 10 related metabolic pathways， including glycerophospholipids and primary bile acid biosynthesis. The quantitative data of targeted metabolomics for SCFAs showed that Dictamni Cortex intervention disrupted the level of propionic acid， butyric acid， acetic acid， caproic acid， isobutyric acid， isovaleric acid， valeric acid， and isocaproic acid in the ileal contents of rats. Compared with those in the control group， the level of isobutyric acid， isovaleric acid， and valeric acid were significantly increased， while the level of propionic acid， butyric acid， and acetic acid were significantly decreased in the ileal contents of rats after Dictamni Cortex intervention （P<0.05， P<0.01）. ConclusionDictamni Cortex can induce intestinal damage by regulating glycerophospholipid metabolism， primary bile acid biosynthesis， and metabolic pathways for SCFAs.