ObjectiveTo investigate the mechanism of Xiezhuo Jiedu prescription in the treatment of ulcerative colitis （UC） by inhibiting ferroptosis and alleviating intestinal mucosal injury based on the nuclear factor E₂ related factor 2/solute carrier family 7 member/glutathione peroxidase 4 （Nrf2/SLC7A11/GPX4） signaling pathway. MethodsA total of 60 male SD rats were divided into a normal group， a model group， high- and low-dose Xiezhuo Jiedu prescription groups （26.64 and 13.32 g·kg^-1， respectively）， a ferroptosis inhibitor group （Ferrostatin-1， 0.005 g·kg^-1）， and a mesalazine group （0.27 g·kg^-1）， with 10 rats in each group. A UC rat model was established by intrarectal administration of trinitrobenzene sulfonic acid （TNBS）-ethanol. The normal group and the model group were intragastrically administered normal saline. The other groups were given intragastric administration according to the corresponding dosage for 7 d. The general condition， disease activity index （DAI） score， colon length， and mucosal injury index （CDMI） score were observed in each group. The pathological changes of colon tissue in each group were observed by hematoxylin-eosin （HE） staining. The intestinal mucosa and mitochondrial morphology in each group were observed by transmission electron microscopy. The expression levels of Occludin， Claudin-1， mucin 2 （MUC2）， and E-cadherin in intestinal tissue were detected by immunofluorescence （IF）. Enzyme-linked immunosorbent assay （ELISA） was used to detect the expression levels of serum tumor necrosis factor-α （TNF-α）， interleukin-6 （IL-6）， and interleukin-10 （IL-10） in each group， and a lactic acid assay kit or ELISA was employed to detect the expression levels of reactive oxygen species （ROS）， ferrous ions （Fe²⁺）， glutathione （GSH）， malondialdehyde （MDA）， 4-hydroxynonenal （4-HNE）， diamine oxidase （DAO）， and D-lactate （D-LA）. Real-time quantitative polymerase chain reaction （Real-time PCR） was applied to detect the mRNA expression levels of Nrf2， SLC7A11， GPX4， Occludin， Claudin-1， MUC2， and E-cadherin in each group， and Western blot was adopted to detect the protein expression levels of Nrf2， p-Nrf2， SLC7A11， and GPX4 in each group. ResultsCompared with the normal group， rats in the model group exhibited listlessness， sluggish response， and mucopurulent and bloody stools. The model group also showed significantly increased DAI score， colon length， CDMI score， and expression levels of TNF-α， IL-6， ROS， Fe²⁺， MDA， 4-HNE， DAO， and D-LA （P<0.01）. In addition， it presented significantly decreased IF values of Occludin， Claudin-1， MUC2， and E-cadherin and mRNA and protein expression levels of IL-10， GSH， Nrf2， p-Nrf2， SLC7A11， and GPX4 （P<0.01）. There were different degrees of improvement in each administration group after treatment， and the improvement was the most significant in the high-dose Xiezhuo Jiedu prescription group （P<0.01）. ConclusionXiezhuo Jiedu prescription may alleviate intestinal mucosal injury by inhibiting ferroptosis of intestinal epithelial cells via regulating the Nrf2/SLC7A11/GPX4 signaling pathway， thereby exhibiting efficacy in the treatment of UC.

ObjectiveThis study aims to explore the potential mechanism of the Xiezhuo Jiedu formula in regulating pyroptosis for the treatment of ulcerative colitis （UC） using bioinformatics and in vivo animal experiments. MethodsDifferentially expressed genes （DEGs） in colon tissues of UC patients were retrieved from the Gene Expression Omnibus （GEO） database. Pyroptosis-related genes were obtained from the GEO and GeneCards databases. The intersection of these datasets yielded pyroptosis-related DEGs （Pyro-DEGs）. Pyro-DEGs were subjected to Gene Ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） enrichment analysis using the Metascape database. A protein-protein interaction （PPI） network was constructed using the STRING database. Least absolute shrinkage and selection operator （LASSO） prediction model and receiver operating characteristic （ROC） analysis were conducted to identify core Pyro-DEGs with diagnostic and therapeutic potential. Immune infiltration analysis of the UC datasets was performed using the deconvolution method （CIBERSORT）， along with correlation analysis with core Pyro-DEGs. Sixty male Sprague-Dawley （SD） rats were randomly divided into a control group， a model group， high-， medium-， and low-dose groups of Xiezhuo Jiedu formula （26.64， 13.32， 6.66 g·kg^-1）， and a mesalazine group （0.27 g·kg^-1）， with 10 rats in each group. UC was established by intrarectal administration of 3，5-trinitrobenzenesulfonic acid （TNBS） dissolved in ethanol. The control and model groups were given distilled water by gavage， while the treatment groups were administered the corresponding drugs for 7 consecutive days. Hematoxylin-eosin （HE） staining was used to observe the colon histopathology. Enzyme-linked immunosorbent assay （ELISA） was used to detect the levels of inflammatory factors such as interleukin-1β （IL-1β）， IL-10， IL-18， and transforming growth factor-β （TGF-β）. Immunohistochemistry （IHC） and Western blot were applied to detect the expression of Caspase-1， gap junction alpha-1 protein （GJA1）， peroxisome proliferator-activated receptor gamma （PPARG）， and S100 calcium-binding protein A8 （S100A8）. Real-time quantitative polymerase chain reaction （Real-time PCR） was utilized to measure mRNA expression of Caspase-1， GJA1， PPARG， and S100A8. Western blot was performed to assess protein expression levels of Caspase-1， GJA1， PPARG， and S100A8. ResultsGEO datasets GSE87466 and GSE87473 yielded 64 Pyro-DEGs. KEGG analysis indicated that these genes were enriched in the NOD-like receptor signaling pathway， tumor necrosis factor （TNF） signaling pathway， and hypoxia-inducible factor 1 （HIF-1） signaling pathway. Four core Pyro-DEGs （Caspase-1， GJA1， PPARG， and S100A8） were identified. Immune infiltration analysis showed that expression of these genes was positively correlated with mast cells， neutrophils， M0 macrophages， M1 macrophages， and dendritic cells. Animal experimental results indicated that compared with the control group， the model group had significantly increased levels of IL-1β and IL-18， significantly decreased levels of IL-10 and TGF-β. The model group showed enhanced Caspase-1， GJA1， and S100A8 staining， and significantly increased mRNA and protein expression of Caspase-1， GJA1， and S100A8 （P<0.01）. In contrast， the expression of PPARG was reduced in the model group （P<0.01）. After treatment， all dosage groups showed varying degrees of improvement （P<0.05， P<0.01）， with the high-dose group showing the most significant improvement （P<0.01）. ConclusionCaspase-1， GJA1， PPARG， and S100A8 are core Pyro-DEGs closely associated with the pathogenesis of UC. These genes may collaborate with immune cells such as mast cells， neutrophils， and M0 macrophages to mediate disease development. The Xiezhuo Jiedu formula may regulate the expression of core Pyro-DEGs through the NOD-like receptor， TNF， and HIF-1 core signaling pathways， thereby modulating immune homeostasis in UC rats and effectively alleviating UC.

Objective:To investigate the relationship between serum annexin A1 (ANXA1), macrophage inflammatory protein 1-α (MIP-1α), soluble triggering receptor expressed on myeloid cells-1 (sTREM-1) and pulmonary infection in elderly patients with coronary heart disease complicated with heart failure and their predictive value.Methods:A total of 197 elderly patients with coronary heart disease combined with heart failure admitted to the Second Hospital of Qinhuangdao from January 2021 to January 2023 were retrospectively selected, and divided into pulmonary infection group (36 cases) and non pulmonary infection group (161 cases) according to whether the patients had pulmonary infection during hospitalization. Serum ANXA1, MIP-1α and sTREM-1 levels were detected in two groups. Multivariate logistic regression model was used to analyze the influencing factors of lung infection in elderly patients with coronary heart disease combined with heart failure. The predictive value of serum ANXA1, MIP-1α and sTREM-1 levels on lung infection in elderly patients with coronary heart disease combined with heart failure was analyzed by receiver operating characteristic (ROC) curve.Results:The incidence of pulmonary infection in 197 elderly patients with coronary heart disease combined with heart failure was 18.27%(36/197). Compared with the non pulmonary infection group, the pulmonary infection group had higher levels of serum ANXA1, MIP-1 α, sTREM-1, C-reactive protein, procalcitonin, and higher proportion of New York Heart Association (NYHA) heart function grade Ⅳ and diabetes, and lower left ventricular ejection fraction (all P<0.05). Multivariate logistic regression analysis showed that NYHA cardiac function grade Ⅳ, diabetes mellitus and elevated levels of procalcitonin, ANXA1, MIP-1α and sTREM-1 were independent risk factors for pulmonary infection in elderly patients with coronary heart disease combined with heart failure (all P<0.05). ROC curve analysis showed that the area under the curve predicted by serum ANXA1, MIP-1α and sTREM-1 combined was 0.909, which was larger than that predicted by serum ANXA1, MIP-1α and sTREM-1 alone. Conclusions:Elevated levels of serum ANXA1, MIP-1α and sTREM-1 are independent risk factors for pulmonary infection in elderly patients with coronary heart disease combined with heart failure, and can be used as auxiliary predictors of pulmonary infection in elderly patients with coronary heart disease combined with heart failure.

Objective:To introduce a new technique of precise exposure and preservation of membranous semicircular canal under endoscope, and to evaluate the protective effect of this method on hearing preservation of semicircular canal occlusion.Methods:The study included three patients with unilateral refractory Meniere′s disease, including 1 male and 2 females, with hearing stage 4 in 2 cases and stage 3 in 1 case, the average hearing thresholds were 72.5, 82.5 and 48.8 dBHL, respectively. All of them were treated with triple semicircular canal occlusion under continuous irrigating mode of endoscopic ear surgery (CIM-EES) in RenMin Hospital of WuHan University. The surgical procedure involved the following key steps: the bony walls of the three semicircular canals near the ampullae were gradually thinned to expose the blue line. The endoscope was positioned as close as possible to the operating area to obtain a clear, magnified view. A micro hook needle was used to create a window on the bony semicircular canals. The hook needle progressively removed the bony material while preserving the membranous canals, which were clearly visible during the procedure. The bone dust was removed out of the surgical area by the continuous exchanged insuline without the help of suction that can avoid the damage of the membranous semicircular canals caused by aspiration and the open bony canals were precisely filled with bone wax and covered with bone powder to ensure complete occlusion of the three semicircular canals. Postoperative follow-up included regular assessments of vertigo control and hearing preservation.Results:All the 3 patients obtained complete hearing preservation at 6 months after surgery, and the hearing preservation rate was 100%. The average hearing thresholds at 6 months postoperatively were 66.3, 81.3, and 50.0 dBHL, respectively. Postoperatively, all patients experienced horizontal spontaneous nystagmus toward the healthy side on the day of surgery and the first day after surgery. One patient reported complete disappearance of tinnitus, while the other two patients showed no change in tinnitus severity. Postoperative MRI revealed complete occlusion of the three semicircular canals with normal vestibular imaging. At 6 months postoperatively, none of the patients experienced any vertigo attacks, suggesting effective control of vertigo symptoms.Conclusions:Precise Semicircular canal occlusion under CIM-EES is a promising technique for the treatment of intractable vertigo in patients with refractory Meniere′s disease. This method provides clear visualization of the membranous canals, reducing the risk of inner ear damage and preserving hearing function of the patients.

Objective:To analyze the blood-transition prototype components and metabolites of Fengliaoxing Fengshi Dieda medicinal liquor.Methods:Ultra-high performance liquid chromatographyquadrupole/electrostatic field orbital trap high resolution mass spectrometry (UHPLC-Q Exactive Focus MS/MS) technique was used to compare the chromatogram differences of Fengliaoxing Fengshi Dieda medicinal liquor extract, blank serum and drug-containing serum. According to the retention time, relative molecular weight and the ratio with primary and secondary ion fragments provided by MS, the prototype components and metabolites of Fengliaoxing Fengshi Dieda medicinal liquor extract were analyzed in serum of rats after oral administration. The detection conditions were as follows: the mobile phase of methanol (A)-0.1% formic acid solution (B) for elution gradient (0-5 min, 5%A; 5-60 min, 5%-95%A; 60-65 min, 95%A), the flow rate of 0.3 ml/min, heated electrospray ionization, detection range of m/z 80-1 200, positive and negative ion scanning modes.Results:A total of 31 transitional components were detected in the serum, of which 9 were prototype components and 22 were metabolites. The 9 prototype components were identified as phenylacetaldehyde, baogongteng C/ erycibellin, p-coumaric acid, 5-Hydroxymethylfurfural, quinic acid, paeonol, 3-Hydroxybenzaldehyde, salicylic acid, and isourecumenol. The 22 metabolites mainly consist of 11 organic acid components, 3 indole components, 2 organic phenolic components, 2 alkaloid components, 1 nucleoside component, 1 amino acid component, 1 lactone component, and 1 sulfonic component. The metabolic pathways were mainly glucuronidation, sulfation and others, which by phase Ⅱ metabolism.Conclusion:Organic phenols and organic acids are the main components that enter the body of Fengliaoxing Fengshi Dieda Medicinal Liquor, while alkaloid compounds and organic acid components may be potential active ingredients for its pharmacological effects.

Objective:To introduce a new design of super-micro flap for endoscopic ear surgery, and to evaluate the application effect of super-micro flap in endoscopic tympanoplasty. Methods:Between January, 2023 and March, 2024, 58 patients（64 ears） with tympanosclerosis underwent tympanoplasty with super-micro flap. Continuous irrigating mode endoscopic ear surgery（CIM-EES） was used to complete type Ⅱ or Ⅲ tympanoplasty with the tragus cartilage with followed up for 12 to 24 months. The operation time, postoperative efficacy and complications were statistically analyzed. Results:Of the 64 ears, 63 ears had primary healing of the tympanic membrane, and 1 ear had cartilage necrosis due to multi-drug resistant bacteria infection. The second operation was performed one year later, and the success rate of operation was 98.40%. The average operation time was （48.40±8.86） minutes. The average hearing threshold of 0.5 kHz to 4.0 kHz before operation was （59.63±10.62） dB HL, and the average air conduction threshold of 0.5 kHz to 4.0 kHz one year after operation was（38.79±10.91） dB HL, which was significantly improved compared with that before operation（P<0.01）. Bone conduction threshold also improved significantly （24.49±8.55） dB HL vs（21.88±7.58） dB HL（P<0.01）. No outer tympanic membrane healing and ear canal scar stenosis occurred. Conclusion:The design of super-micro flap can effectively solve the interference of flap floating during continuous irrigating mode in endoscopic ear surgery, relieve the difficulty of flap reposition, simplify the operation process, help to shorten the operation time, and reduce the possibility of circular scar stenosis of conventional free flap, which provides a new flap design option for endoscopic ear surgery.
Humans ; Tympanoplasty/methods* ; Endoscopy/methods* ; Surgical Flaps ; Male ; Female ; Adult ; Myringosclerosis/surgery* ; Middle Aged ; Treatment Outcome ; Tympanic Membrane/surgery* ; Adolescent ; Young Adult

Objective:To assess the humanistic caring ability of clinical interns, analyze its influencing factors, explore the correlation between humanistic caring ability and narrative ability, and provide a basis for formulating narrative medical education scheme for clinical interns.Methods:A total of 163 clinical interns from the Class of 2020 completing internships at a Grade A tertiary hospital in Zhuhai were enrolled as research subjects using convenience sampling. A questionnaire survey was conducted using a baseline demographic survey, a humanistic caring ability inventory, and a medical narrative ability scale. Pearson correlation analysis and multiple linear regression analysis were conducted to explore the influencing factors for humanistic caring ability and its correlation with narrative ability.Results:The overall score of humanistic caring ability was (187.52±20.30). There were significant differences between groups in self-evaluation of humanistic caring ability, degree of satisfaction with the major, and relationship with classmates ( P<0.05). There was a positive correlation between narrative ability and caring ability in both total score and individual dimensions ( r=0.600, P<0.01). After adjusting for demographic differences, relationship with classmates ( β=0.138, P=0.042), attentive listening ( β=0.354, P<0.01), and reflective representation ( β=0.259, P=0.008) exhibited a significant positive impact on humanistic caring ability. However, this impact was not observed for the understanding-response dimension. Conclusions:The humanistic caring ability of clinical interns is low and their medical narrative ability is positively correlated with their humanistic caring ability. Therefore, narrative medical education should focus on training medical students' attentive listening and reflective representation to enhance humanistic literacy and promote their career development.

Objective:To assess the humanistic caring ability of clinical interns, analyze its influencing factors, explore the correlation between humanistic caring ability and narrative ability, and provide a basis for formulating narrative medical education scheme for clinical interns.Methods:A total of 163 clinical interns from the Class of 2020 completing internships at a Grade A tertiary hospital in Zhuhai were enrolled as research subjects using convenience sampling. A questionnaire survey was conducted using a baseline demographic survey, a humanistic caring ability inventory, and a medical narrative ability scale. Pearson correlation analysis and multiple linear regression analysis were conducted to explore the influencing factors for humanistic caring ability and its correlation with narrative ability.Results:The overall score of humanistic caring ability was (187.52±20.30). There were significant differences between groups in self-evaluation of humanistic caring ability, degree of satisfaction with the major, and relationship with classmates ( P<0.05). There was a positive correlation between narrative ability and caring ability in both total score and individual dimensions ( r=0.600, P<0.01). After adjusting for demographic differences, relationship with classmates ( β=0.138, P=0.042), attentive listening ( β=0.354, P<0.01), and reflective representation ( β=0.259, P=0.008) exhibited a significant positive impact on humanistic caring ability. However, this impact was not observed for the understanding-response dimension. Conclusions:The humanistic caring ability of clinical interns is low and their medical narrative ability is positively correlated with their humanistic caring ability. Therefore, narrative medical education should focus on training medical students' attentive listening and reflective representation to enhance humanistic literacy and promote their career development.

Objective:To investigate the molecular mechanism by which 2',4'-dihydroxychalcone(D2)inhibits proliferation,migration,and epithelial-mesenchymal transition(EMT)in colorectal cancer cells through miR-7-5p-mediated autophagy.Methods:Human colorectal cancer cell lines HCT-15 and SW620 were treated with D2 at concentrations of 0,12.5,25,and 50 μmol/L.Cell proliferation and clonogenic capacity were evaluated using MTT and colony formation assays.Cell migration was assessed by wound healing and Transwell assays.WB assay was used to detect the expression of EMT-related proteins,autophagy-related proteins,and key components of the PI3K/AKT/mTOR pathway.Autophagosome formation was visualized by immunofluorescence staining.TCGA database and KEGG pathway analyses were performed to evaluate miR-7-5p expression and its association with colorectal cancer.RT-qPCR was used to quantify miR-7-5p expression,and lentiviral transduction was employed to establish stable miR-7-5p knockdown or overexpression cell lines.Results:D2 significantly inhibited colorectal cancer cell proliferation,migration,and EMT(P<0.05 or P<0.01).TCGA and KEGG analyses revealed that miR-7-5p expression was downregulated in colorectal cancer and closely associated with disease progression.D2 treatment(12.5,25,and 50 μmol/L)significantly upregulated miR-7-5p expression in HCT-15 and SW620 cells(P<0.01).At 25 μmol/L,D2 increased the expression of autophagy-related proteins(LC3 and p-ULK1)and inhibited the PI3K/AKT/mTOR signaling pathway(P<0.05),accompanied by increased autophagosome formation(P<0.01).In miR-7-5p-knockdown cells treated with D2,the levels of LC3 and p-ULK1 were significantly reduced compared to D2-only treated cells(P<0.05 or P<0.01).Conclusion:D2 upregulates miR-7-5p to induce autophagy,thereby inhibiting colorectal cancer cell proliferation,migration,and EMT,possibly through suppression of the PI3K/AKT/mTOR signaling pathway.

OBJECTIVE To develop a highly sensitive and specific quantitative real-time poly-merase chain reaction(qPCR)method for detecting the VGM-R02b(a gene therapy drug for glutaric acidemia type Ⅰ)gene in cynomolgus monkeys and analyze the biological distribution of VGM-R02b.METHODS A standard curve was constructed using the VGM-R02b standard plasmid[an adeno-associ-ated virus serotype 9(AAV9)capsid]on a qPCR platform.The detection method was optimized and validated for key parameters,including the quantitative range,accuracy,precision,dilution linearity,selectivity,specificity,stability,and parallelism.The established method was used to determine the target gene of VGM-R02b in the blood,brain,stomach,heart,liver,spleen,lung,kidney,thymus,and duodenum of cynomolgus monkeys on day 29(D29)and D92 after a single,unilateral intraventricular injection of VGM-R02b.The biodistribution of VGM-R02b in cynomolgus monkeys was analyzed.RESULTS A qPCR method for quantifying the VGM-R02b target gene in cynomolgus monkeys was established and validated.The standard curve demonstrated a quantitative range of 5.00×109 to 5.00×10^1 copies·μg-1 DNA,with excellent precision,accuracy,dilution linearity,selectivity,and specificity.The target gene in tissues remained stable after being stored at room temperature for 4 h,-15 to-25℃ for 9 d,-60 to-80℃ for 90 d and after five freeze-thaw cycles.Similarly,the target gene in whole blood remained stable after being stored at room temperature for 4 h,-15 to-25℃ for 93 d,-60 to-80℃ for 93 d and after five freeze-thaw cycles.Extracted nucleic acid samples also showed stability after being stored at room temperature for 4.25 h,2-8℃ for 24.16 h,-60 to-80℃ for 109 d and after five freeze-thaw cycles.The method was also applied to evaluate the biological distribution of the target gene in cynomolgus monkeys.In the control group,the target gene was undetectable on D29 and D92 post-administration,but in the drug administration group,the target gene was distributed across the tissues,with higher concen-trations observed in the brain,liver,spleen,and spinal cord,and there were no significant differences in the target gene content across the tissues between D29 and D92.CONCLUSION The established qPCR method is robust,reliable and suitable for determination of VGM-R02b target gene in cynomolgus monkeys.