ObjectiveTo establish a reliable chronic obstructive pulmonary disease (COPD) mouse model based on a self-developed multichannel automatic control system for long-term continuous cigarette smoke exposure in small animals using a novel continuous cigarette smoke exposure method, and to conduct phenotypic evaluation and analysis, thereby providing an animal experimental basis for investigating COPD pathogenesis and prevention strategies. MethodsTwenty male C57BL/6J mice aged 6 weeks were randomly and equally divided into a control group and a model group. The model group (n=10) underwent 6 h of continuous cigarette smoke exposure daily (6 cigarettes per day for 12 consecutive weeks), while the control group (n=10) received no intervention. Body weight was monitored biweekly. Post-exposure, in vivo micro-CT imaging was performed. After euthanasia, serum and bronchoalveolar lavage fluid (BALF) levels of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were quantified by ELISA. Lung tissues underwent H&E and Masson's trichrome staining to observe changes in lung morphology and inflammatory cell infiltration, and the mean linear intercept (MLI) was calculated, thereby comprehensively evaluating the clinical features of COPD in the mouse model. ResultsCompared with the control group, the model group showed significantly reduced body weight (P<0.01) from the fourth week. Compared with the control group, IL-6 level in the serum and BALF of the model group increased by 27.2% and 140.0%, respectively (P<0.01). TNF-α level in the serum and bronchoalveolar lavage fluid of the model group increased by 16.7% (P<0.01) and 19.3% (P<0.05), respectively. Histopathological examination revealed alveolar wall thinning, septal rupture, emphysematous bullae formation, reduced alveolar count, bronchial wall thickening with lumen narrowing, and inflammatory cell infiltration. MLI was significantly elevated (P<0.01). Masson's staining confirmed collagen deposition and bronchial remodeling. Micro-CT demonstrated localized high-density shadows exhibiting typical features of chronic bronchitis. Conclusion The self-developed device enables long-term continuous smoke exposure, and the successfully established COPD mouse model exhibits pathological features highly consistent with clinical manifestations, offering an efficient and reliable tool for COPD research.

ObjectiveTo explore the value of a multimodal MRI-based radiomics nomogram for differentiating human epidermal growth factor receptor-2 （HER-2） negative breast cancer molecular subtypes.MethodsA retrospective analysis was conducted on 190 patients with HER-2 negative breast cancer who underwent multimodal MRI examination， and the patients were divided into two molecular subtype groups： a HER-2 low expression group （n=108） and a HER-2 zero expression group （n=82）. The cases were randomly stratified and sampled at a ratio of 7∶3 and divided into a training set of 133 cases and a testing set of 57 cases. The clinical and radiological features of the patients were collected， the radiomics features based on T2-weighted imaging （T2WI）， diffusion-weighted imaging （DWI）， and dynamic contrast-enhanced （DCE）-MRI were extracted， and the clinical-radiological model， unimodal radiomics model， multimodal radiomics model， and combined model were constructed respectively. Then the nomogram combined multimodal radiomics signature （radsocre） with clinical-radiological features was used to construct a visualized predictive model， and the area under the curve （AUC） was used to compare the effectiveness of different models in distinguishing HER-2 low expression and zero expression subtypes.ResultsA significant difference in radscore was demonstrated between the HER-2 low and HER-2 zero expression groups in both the training （P<0.000 1） and testing sets （P<0.01）. The AUC of the multimodal radiomics model in the training set and the testing set were 0.914 and 0.836， respectively， which was superior to any unimodal radiomics model. The nomogram demonstrated great diagnostic efficacy （AUC=0.930 in training set； AUC=0.865 in testing set）.ConclusionA multimodal MRI-based nomogram incorporating radsocre and clinical-radiological features can accurately distinguish the subtypes of HER-2 negative breast cancer.

ObjectiveTo investigate the effect of Zishen Huoxue Formula （ZSXHF） on molecular-targeted therapy-associated proteinuria in patients with primary liver cancer （PLC）， to assess the efficacy of ZSXHF in the treatment of molecular-targeted therapy-associated proteinuria， and to provide a basis for clinical medication. MethodsA retrospective cohort study was conducted among the PLC patients with molecular-targeted therapy-associated proteinuria who were diagnosed and treated in The Department of Hepatology of Chinese PLA General Hospital， from January 1， 2022 to July 1， 2025. With ZSXHF treatment as the exposure factor， the patients with a cumulative treatment duration of ≥9 weeks were enrolled as traditional Chinese medicine （TCM） group， while those without TCM treatment were enrolled as control group. Propensity score matching was performed for the two groups at a ratio of 1∶1 based on sex， age， 24-hour urinary protein， blood urea nitrogen， and serum creatinine. The independent-samples t test was used for comparison of normally distributed continuous data between two groups， and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups； the chi-square test was used for comparison of categorical data between groups. Univariate and multivariate Logistic regression analyses were used to investigate the influencing factors for promoting the improvement of targeted-therapy-associated proteinuria. ResultsA total of 137 PLC patients with targeted-therapy-associated proteinuria were enrolled， with 34 patients in the TCM group and 103 in the control group. After follow-up for 6 months， the TCM group had a significant improvement in urinary protein grade compared with the control group （χ²=9.261， P=0.016）. There were 25 patients in each group after propensity score matching， and after follow-up for 6 months， there were significant differences between the two groups in urinary protein grade （χ²=15.689， P<0.001） and 24-hour urinary protein （Z=-3.075， P=0.002）. After cumulative treatment with ZSXHF for ≥9 weeks， the TCM group had a significantly greater change in 24-hour urinary protein from baseline compared with the control group （t=-2.514， P=0.016）， while there were no significant differences in the changes in liver and renal function after ZSXHF intervention between the two groups （all P>0.05）. The multivariate Logistic regression analysis showed that ZSXHF treatment （odds ratio=2.901， 95% confidence interval： 1.135 — 7.417， P=0.026） was an independent influencing factor for improvement in molecular-targeted therapy-associated proteinuria. ConclusionZSHXF can effectively alleviate molecular-targeted therapy-associated proteinuria in PLC patients with a favorable safety profile， which provides a new reference for TCM prevention and treatment of molecular-targeted therapy-associated adverse reactions in PLC patients.

Objective To explore the effects of long non-coding RNA(LncRNA)metastasis associated in lung denocarcinoma transcript 1(MALAT1)on high glucose-induced cardiomyocyte injury by regulating miR-320-3p-protein kinase B3(Akt3)axis.Methods H9c2 cells were divided into normal control(NC)group and high glucose(HG)group.After transfection,the cells were cultured in HG group were further divdided into subgroups as follows:HG+MALAT1 knock-down(HG+sh-MALAT1)group,HG+sh-NC group,HG+miR-320-3p mimic group,HG+mimic-NC group,HG+sh-MALAT1+miR-320-3p inhibitor group and HG+sh-MALAT1+pcDNA3.1-Akt3 group.Cell activity,apoptosis rate,B-cell lymphoma 2-related protein X(Bax),B-cell lymphoma factor 2(Bcl-2),cleaved caspase-3,Akt3 protein,MALAT1,miR-320-3p,Akt3 mRNA expression,interleukin(IL-1β),IL-6,tumor necrosis factor-α(TNF-α)were compared among the groups.Results The apoptosis rate of cardiomyocytes,the expressions of IL-1β,IL-6,TNF-α,Bax,cleaved caspase-3/caspase-3 protein,MALAT1 and Akt3 mRNA increased(P<0.05),while the expression of Bcl-2 protein and miR-320-3p decreased in HG group(P<0.05).After knocking down LncRNA MALAT1,apoptosis rate of HG-induced cardiomyocytes and levels of inflammatory factors were decreased(P<0.05).MALAT1 could negatively regulate expression of miR-320-3p,and miR-320-3p could negatively regulate expression of Akt3 protein.The overexpression of miR-320-3p could decrease apoptosis rate and inflammation level,while overexpression of Akt3 or inhibition of miR-320-3p expression could increase apoptosis rate of cardiomyocytes and inflammation level(P<0.05).Conclusions LncRNA MALAT1 can promote the expression of Akt3 protein by inhibiting miR-320-3p expression,thereby reduce activity of HG-induced cardiomyocytes and aggravate cardiomyocytes injury.

Gene-edited mouse models,as a core tool in modern biomedical research,play an irreplaceable role in fields such as disease mechanism analysis,drug development,and gene function studies.However,universities often face challenges including technical complexity,dispersed resources,and low management efficiency during the preparation,breeding,and data analysis of gene-edited mice.To address these issues,establishing a one-stop service platform for gene-edited mice that integrates full-chain technical services and an intelligent management system has become an inevitable choice for university laboratory animal centers to break through current difficulties.This platform must possess core functions including gene-edited mouse construction,breeding and colony expansion,biological purification,strain preservation,and endangered strain rescue,forming a whole-process technical service system covering"target design-model construction-population expansion-genetic quality control-biological sample preservation",to promote the strategic transformation of university laboratory animal centers from providing"single technical services"to providing"whole-process solutions".Taking the Laboratory Animal Center of Shanghai Jiao Tong University as an example,through technical optimization,the technical defects such as low microoperation success rate and insufficient standardization of genotype identification were solved.Through traceability management and information sharing,problems such as chaotic management of mouse strains in research groups and waste of research funds caused by repeated constructions were solved.Through systematic integration and optimization of technical services,problems such as low efficiency,resource waste,and collaboration difficulties that may be caused by fragmented service systems were solved.This paper systematically discusses the construction path and practical experience of the gene-edited mouse service platform from three perspectives:intensive management,service capacity building,and support system construction,aiming to provide referable management models and implementation paths for the construction of similar platforms in other universities.

Objective:To establish the reference interval of 12 types of cytokines(IL-1β,IL-2,IL-4,IL-5,IL-6,IL-8,IL-10,IL-12p70,IL-17,IFN-γ,IFN-α,TNF-α)in adult plasma based on multiple microsphere flow immunofluorescence(MBFFI).Methods:A total of 140 healthy adult patients who were examined at Xuzhou Central Hospital between January 2022 and December 2023 were included in the study.Plasma cytokine levels were detected and reference intervals were established by the flow cytometer and the assay kits produced by Qingdao Raisecare Biotechnology Co.,Ltd and Jiangsu BioPredia Biotechnology Co.,Ltd.Results:All of the cytokines exhibited a non-normal distribution,and there was a discrepancy in the 95％reference interval between the two re-agents.The reference intervals for the 12 cytokine kits produced by Qingdao Raisecare Biotechnology Co.,Ltd.were as follows:IFN-α:<4.91 pg/ml,IL-12 p70:<1.95 pg/ml,IL-5:<12.72 pg/ml,IL-8:<60.68 pg/ml,IL-1β:<27.67 pg/ml,IL-2:<5.01 pg/ml,IL-4:<1.22 pg/ml,IL-6:<6.11 pg/ml,TNF-α:<2.92 pg/ml,IL-17:<10.27 pg/ml,IL-10:<6.88 pg/ml,IFN-γ:<17.68 pg/ml.The reference intervals of the 12 cytokines produced by Jiangsu BioPredia Biotechnology Co.,Ltd.were as follows:IFN-α:<4.05 pg/ml,IL-12 p70:<7.33 pg/ml,IL-5:<7.80 pg/ml,IL-8:<13.24 pg/ml,IL-1β:<19.24 pg/ml,IL-2:<2.42 pg/ml,IL-4:<0.99 pg/ml,IL-6:<2.10 pg/ml,TNF-α:<0.87 pg/ml,IL-17:<1.42 pg/ml,IL-10:<1.10 pg/ml,IFN-γ:<1.34 pg/ml.Conclusion:In this study,the ref-erence range of two reagents for the detection of 12 kinds of cytokines in plasma of healthy adults is established by MBFFI,which pro-vides a valuable reference for the diagnosis and treatment of clinical-related diseases.

Objectives:To investigate the risk factors and treatment methods for distal junctional isthmic spondylolisthesis(DJIS)following posterior lumbar decompression and fixation surgery.Methods:The 10 patients who were treated at our hospital for DJIS following posterior decompression and fixation between January 2015 and January 2022 were retrospectively analyzed.The patients were included in the DJIS group,including 7 males and 3 females,aged 63.4±10.3(45-75)years old.And according to age,gender,preoperative diagnosis,operative stage,and operative method,the patients were matched in a ratio of 1∶2 with some other patients who didn't develop DJIS after underwent posterior decompression and fixation at our hospital due to lumbar degenerative diseases during the same period as control(20 cases).The general data[body mass index(BMI),L1 CT value,proportion of patients with osteoporosis)],laminectomy range of the lower vertebra(transverse decompression percentage of lamina,spinous process resection percentage),pelvic incidence(PI),and postoperative lumbar lordosis(LL),pelvic tilt(PT),sacral slope(SS),etc.of the two groups were compared to explore the risk factors for DJIS after posterior lumbar decompression and fixation.The treatment methods for DJIS were also summarized.Results:The BMI of patients in the DJIS group was significantly higher than that of the control group(27.4±4.1kg/m2 vs.23.7±3.4kg/m2,P＜0.001).The L1 vertebral CT value of the DJIS group was significantly lower than that of the control group(105.2±43.9HU vs.133.5±23.5HU,P=0.028),and the proportion of patients with osteoporosis of the DJIS group was higher(70％vs.10％,P=0.003).The DJIS group was greater in PI(52.5°±8.8° vs.45.8°±7.4°,P＜0.05)and postoperative LL(47.4°±14.3° vs.36.5°±10.6°,P＜0.05)significantly than the control group,PT and SS were not significantly different between the two groups(P＞0.05).Additionally,the transverse decompression percentage of lamina of the lower vertebra[(89.3±9.0)％vs.(78.0±3.2)％,P＜0.05]and the spinous process resection percentage of the distal vertebra[(51.1±16.1)％vs.(39.3±9.1)％,P＜0.05]in the DJIS group were also significantly higher than those in the control group.Eight DJIS patients underwent distal decompression,reduction,fixation,and fusion surgery,and their quality of life scores significantly improved after revision surgery.Two DJIS cases with mild clinical manifestations were treated conservatively,no symptom exacerbation was reported during follow-up.Conclusions:High BMI,osteoporosis,high PI,and excessive distal vertebral lamina resection during surgery are potential risk factors for DJIS after posterior lumbar decompression and fixation.

Objective To explore the mechanism underlying the role of disulfidptosis-related genes(DRGs)in the disease progression of metabolically associated fatty liver disease(MAFLD)based on bioinformatics.Methods In this study,the GEO database was utilized to screen for eligible MAFLD expression data,conduct differential gene analysis,and identify DRGs through consistent clustering to subtype MAFLD patients.The immune infiltration status among subtypes was further evaluated,and the infiltration of immune cells was analyzed using the CIBERSORT algorithm.The gene modules related to the disease were selected through weighted gene co-expression network analysis(WGCNA).Subsequently,a diagnostic model was constructed based on DRGs using machine learning models,and the performance of the model was verified.Finally,the stability of DRGs among different subtypes was evaluated using an external dataset,and the significance of the results was analyzed using statistical tests.Results Through the analysis of the dataset GSE31803,six disulfide death genes,namely,SLC3A2,NCKAP1,CYFIP1,FLNA,MYL6 and MYH10,which were closely related to the clinical characteristics of MAFLD,were screened out.MAFLD patients were classified into two subtypes,with subtype 1 having a higher level of immune cell infiltration.Key gene modules were identified through WGCNA.Through machine learning screening,the support vector machine(SVM)model was determined as the optimal classification model.External validation confirmed the stability and effectiveness of the key genes in different subtypes of MAFLD.Conclusion Based on DRGs,two highly heterogeneous subtypes of MAFLD were identified,which exhibited significant differences in clinical characteristics,biological processes and immune status,indicating that DRGs play a crucial role in the occurrence and development of MAFLD.

Objective To observe changes of white matter function in adolescent smoker(AS)with resting-state functional MRI(rs-fMRI)fractional amplitude of low-frequency fluctuation(fALFF)technique.Methods Forty-five adolescents(AS group)and 45 control subjects(control group)were prospectively enrolled,and brain rs-fMRI were acquired.Brain regions with fALFF being different between groups were observed,and the correlations with clinical indicators were analyzed.Results Compared with that in control group,fALFF of the right superior longitudinal fasciculus significantly elevated in AS group(FDR correct Q＜0.05),in which the peak of the cluster was positively correlated with score of Fagerstr?m test for nicotine dependence(FTND)(r=0.294,P=0.049).Conclusion White matter function changed in AS,presenting as significantly increased fALFF in right superior longitudinal fasciculus,which was positively correlated with nicotine dependence.

Objective:To investigate the neuroprotective effects of ginsenoside Rb1 in neonatal mice with Hypoxic Ischemia(HI)and analyze its potential molecular mechanisms.Methods:Seven-day-old C57BL/6 neonatal mice were randomly assigned to three groups:Sham group,hypoxic-ischemic(HI)model group,and HI model+ginsenoside Rb1 intervention group(HI+Rb1),with 10 mice per group.The modified Rice-Vannucci method was used to establish the HI model,and ginsenoside Rb1(20 mg/kg)was administered via intraperitoneal injection for 7 consecutive days post-surgery(once per day).Brain damage was assessed on days 7 and 14 post-surgery by evaluating cortical neurons and glial cell numbers,as well as the activation status of the ERK signaling pathway.Additionally,in utero electroporation(IUE)was used to overexpress the ERK signaling pathway in the cortical neurons,and the impact of ERK activation on glial cell development was observed.Further,IUE was used to overexpress ERK in the cortex of P0 neonatal mice,fol-lowed by the HI model on day 7 to analyze the effects of enhanced ERK signaling on oligodendrocyte development and myelin regeneration.Results:Compared to the HI group,the HI+Rb1 intervention group showed significant improve-ment in motor ability,reduction in brain injury area,less mature neuron loss,and increased newborn neurons.Addi-tionally,the number of oligodendrocytes in the cortex was increased,and the activation of the ERK signaling pathway was enhanced.In mice with overexpression of the ERK signaling pathway in the cortex,there was a significant increase in oligodendrocytes.In the HI model with ERK overexpression,an increased number of oligodendrocyte precursor cells were found around the brain injury area,consistent with the results of ginsenoside Rb1 intervention.Conclusion:Gin-senoside Rb1 exerts neuroprotective effects in neonatal mice with hypoxic-ischemic brain injury,potentially through the enhancement of ERK signaling,promoting oligodendrocyte proliferation and myelin regeneration.