[摘 要] 目的：通过多组学整合分析，解析肠道菌群在胆管癌（CCA）发生发展中的潜在作用机制并识别相关关键基因。方法：基于SRA数据库的16S rRNA测序数据，比较CCA患者与健康对照者的肠道菌群组成；采用孟德尔随机化（MR）分析评估菌群与CCA风险的遗传关联。通过gutMGene和GeneCards数据库获取相关代谢物与基因，进行代谢和功能富集分析。整合GEO单细胞转录组数据（GSE213452），解析肿瘤微环境的细胞组成，重点关注T细胞亚群及其功能状态，并结合TCGA-CHOL数据集验证关键候选基因的表达差异。结果：与健康对照组相比，CCA患者肠道菌群组成发生显著改变，变形菌门下菌群异常富集（LDA > 4）。MR分析进一步证实，肠杆菌目与肠杆菌科的遗传易感性均与CCA风险呈正向关联。代谢通路富集分析提示，菌群相关代谢物主要参与嘌呤代谢及糖酵解/糖异生等通路；功能富集分析显示，相关基因显著富集于NOD样受体、IL-17、Toll样受体及NF-κB信号通路等炎症免疫通路。单细胞转录组分析结果显示，CCA组织中肿瘤细胞比例显著升高（P < 0.05），T细胞比例由20.7%增至39.2%；拟时序分析结果表明，MKI67⁺ T细胞处于分化末期并呈高增殖特征，其差异基因与菌群相关基因存在交集，其中SERPINA1和IFNG表达在肿瘤免疫微环境中显著变化（P < 0.001），可能发挥核心调控作用。TCGA-CHOL数据集验证显示，SERPINA1在CCA肿瘤组织中显著下调（P < 0.001），而IFNG在肿瘤与正常组织间无显著差异（P > 0.05）。结论：肠道菌群失衡（尤其是肠杆菌科异常增殖）可能通过代谢-免疫调控网络促进CCA进展，T细胞功能变化与关键基因（SERPINA1和IFNG）在MKI67+ T细胞中的差异化表达模式密切相关。

low transit constipation （STC） is a common functional intestinal disorder caused by impaired colonic transit function， characterized by reduced bowel movement frequency， hard stools， and difficulty in defecation. The mitogen-activated protein kinase （MAPK） signaling pathway， which mainly includes extracellular signal-regulated kinase （ERK）， c-Jun N-terminal kinase （JNK）， and p38 subtypes， plays a critical regulatory role in the occurrence and development of STC. This paper systematically reviews the multiple pathogenic mechanisms of the MAPK signaling pathway in STC and the research progress of traditional Chinese medicine （TCM） intervention.At the mechanistic level， the MAPK signaling pathway promotes the progression of STC through the following links：（1） Activation of p38 upregulates the expression of aquaporin 3 （AQP3）/AQP4 in the colon， leading to excessive reabsorption of water in the intestinal lumen； （2） It forms a positive feedback loop with nuclear factor-κB （NF-κB） to maintain low-grade intestinal inflammation， releases inflammatory factors such as tumor necrosis factor-α （TNF-α） and interleukin-1β （IL-1β）， and inhibits smooth muscle contraction； （3） Overactivation of p38 downregulates the expression of occludin and mucin 2 while upregulates the expression of claudin-2， thereby disrupting the mucosal barrier； （4） The JNK/p38 signaling pathway activates the caspase cascade to induce apoptosis of intestinal epithelial cells， neurons， and interstitial cells of Cajal； （5） Abnormal ERK signaling and excessive activation of p38/JNK inhibit intestinal smooth muscle contraction and reduce 5-hydroxytryptamine secretion， ultimately resulting in impaired colonic transit function.At the intervention level， TCM compound formulas and single herbs have been proven to improve STC by regulating the MAPK signaling pathway. Their effects are syndrome type-dependent：yin-nourishing formulas （Zengye Chengqi Tang， Tongbian Tang） mainly regulate the ERK/AQP axis； yang-warming formulas （Jichuan Jian） target both ERK/JNK and anti-apoptosis； heat-clearing formulas （Sanren Tang） focus on p38/NF-κB anti-inflammation. A single drug can simultaneously cover multiple aspects including water metabolism， inflammation， barrier function， apoptosis， and intestinal motility.Current relevant studies still have limitations such as mechanisms mostly remaining at the correlational level and a lack of disease-syndrome integrated research models. Future studies should combine specific inhibitors or gene knockout to identify core targets， establish disease-syndrome integrated STC models， and use network pharmacology and molecular docking techniques to deeply analyze the fine mechanism of “component-target-phenotype”， so as to provide high-quality evidence for the precise regulation of the MAPK signaling pathway by TCM in the intervention of STC.

Objective:To evaluate the clinical outcomes of combined complete preservation of chordal structure mitral valve replacement (C-MVR) with total anatomical arterial myocardial revascularization (TACR) in coronary patients with moderate-to-severe or severe ischemic mitral regurgitation (IMR).Methods:This is a retrospective multi-center case series study. Data were retrospectively collected from 127 patients with coronary artery disease with moderate to severe or severe IMR who received TACR with C-MVR from July 2015 to April 2024 in 13 hospitals in China. There were 90 males and 37 females, aged (56.5±10.7) years (range: 33 to 74 years). Perioperative data and follow-up data including left ventricular ejection fraction, left ventricular end-diastolic diameter, and patency rate of arterial grafts of patients were collected. Comparisons were made using paired sample t-test or χ2 test. Results:In this cohort of 127 patients, 67 underwent concurrent tricuspid valve repair. During surgery, 113 grafts of the left internal mammary artery (LIMA), 127 grafts of the left radial artery, 80 grafts of the right radial artery, and 110 grafts of the right internal mammary artery (RIMA) were harvested. The number of the distal anastomosis was 4.2±0.4 (range: 3 to 5). The aortic cross-clamp time and cardiopulmonary bypass time were (97.5±23.4) minutes (range: 90 to 161 minutes) and (145.4±19.2) minutes (range: 101 to 210 minutes), respectively. There was one operative death. Intraoperative placement of an intra-aortic balloon pump was performed in 21 patients to improve the left ventricular ejection. No sternal ischemic occurred. All patients completed follow-up, with a mean follow-up period of (64.3±7.5) months (range: 4 to 110 months). No major cerebrovascular events occurred during the follow-up period, and all patients survived. Left ventricular ejection fraction improved postoperatively (55.0%±5.3% vs. 41.0%±15.3%, t=17.23, P<0.01). The proportion of patients with New York Heart Association functional class ≤2 increased postoperatively (23.6% (30/127) vs. 87.3% (110/126), χ2=103.77, P<0.01). The proportion of patients with Canadian Cardiovascular Society Angina Classification ≤3 decreased postoperatively (4.8% (6/126) vs. 78.7% (100/127), χ2=142.19, P<0.01). The left ventricular end-diastolic diameter decreased postoperatively ((5.70±4.50) cm vs. (6.10±0.23) cm, t=12.15, P<0.01). Coronary multi-detector computed tomography angiography (MDCTA) follow-up was conducted for (60.5±11.7) months (range: 6 to 109 months) postoperatively. MDCTA confirmed the patency rates of the grafts: 96.4% (108/112) for the LIMA grafts, 88.9% (112/126) for the left radial artery grafts, 93.7% (74/79) for the right radial artery grafts, and 90.9% (100/110) for the free RIMA grafts. No significant differences in graft patency rates were observed between the arterial grafts ( χ2=5.24, P=0.155). Conclusion:The results of this multi-centre study demonstrate satisfactory mid-term results of C-MVR with TACR for the treatment of coronary artery disease with moderate to severe or severe IMR.

Objective To investigate the effect of wogonin(WOG)on retinal ganglion cell(RGC)damage in chron-ic glaucoma rats and its relationship with the regulation of the JAK2/STAT3 signaling pathway.Methods A chronic glau-coma rat model was established in SPF-grade healthy male SD rats(50 modeled successfully).Successfully modeled rats were randomly divided into 6 groups(n=10 per group).The model group received an equal volume of normal saline(once daily,10 mL·kg-1 by gavage and 1 mL·kg-1 by intraperitoneal injection).The low-dose WOG group(L-WOG),medium-dose WOG group(M-WOG),and high-dose WOG group(H-WOG)received WOG at 7 mg·kg-1,14 mg·kg-1,and 28 mg·kg-1 by gavage,respectively(once daily,gavage volume 10 mL·kg-1).The high-dose WOG+Colivelin group(H-WOG+Colivelin)received WOG by gavage(28 mg·kg-1,once daily,10 mL·kg-1)plus intraperitoneal injection of the JAK2/STAT3 pathway activator Colivelin(1 mg·kg-1,once daily,injection volume 1 mL·kg-1).An additional 10 normal rats served as the sham-operated control group(Control group),undergoing surgery to expose the superior episcleral vein without cauterization,and receiving normal saline postoperatively in the same manner as the model group.All treatments lasted for 8 weeks.Intraocular pressure was measured using a tonometer.Retinal morphological changes and RGC apopto-sis were assessed by hematoxylin-eosin(HE)staining and TUNEL staining,respectively.Superoxide dismutase(SOD)ac-tivity and malondialdehyde(MDA)content in retinal tissue were measured by ELISA.Protein expression changes in the JAK2/STAT3 signaling pathway in retinal tissue were detected by Western blot.Results Retinal structure was normal in the Control group,with clear layers and densely arranged cells.Compared to the Control group,the model group showed significantly thinner retinal tissue,disorganized structure,sparsely arranged cells,and vacuolation.Compared to the model group,the L-WOG,M-WOG,and H-WOG groups exhibited increased retinal thickness and more orderly cell arrangement.Compared to the H-WOG group,the H-WOG+Colivelin group showed relatively thinner retinal tissue and sparser cell ar-rangement.After intervention,the RGC apoptosis rate in the model group(7.32％±1.07％)was significantly higher than that in the Control group(0.61％±0.12％)(P＜0.05).The apoptosis rates in the L-WOG group(5.67％±0.63％),M-WOG group(3.34％±0.75％),and H-WOG group(1.12％±0.35％)were significantly lower than that in the model group(all P＜0.05),showing a dose-dependent decreasing trend.The apoptosis rate in the H-WOG+Colivelin group(6.83％±1.56％)was significantly higher than that in the H-WOG group(P＜0.05).Compared to the Control group,SOD activity in the retinal tissue of the model group was significantly decreased(P＜0.05),while MDA content was significantly increased(P＜0.05).Compared to the model group,SOD activity was significantly increased and MDA content was significantly de-creased in the L-WOG,M-WOG,and H-WOG groups(all P＜0.05),with this effect being dose-dependent(i.e.,H-WOG group＞M-WOG group＞L-WOG group).Compared to the H-WOG group,the H-WOG+Colivelin group showed signifi-cantly decreased SOD activity and significantly increased MDA content(P＜0.05).Compared to the Control group,the ra-tios of p-JAK2/JAK2 and p-STAT3/STAT3 expression in the retinal tissue of the model group were significantly upregulated(all P＜0.05).Compared to the model group,the ratios of p-JAK2/JAK2 and p-STAT3/STAT3 expression were significantly downregulated in the L-WOG,M-WOG,and H-WOG groups(all P＜0.05).Compared to the H-WOG group,the ratios of p-JAK2/JAK2 and p-STAT3/STAT3 expression were significantly upregulated in the H-WOG+Colivelin group(all P＜0.05).Conclusion WOG may alleviate retinal tissue damage in chronic glaucoma rats by inhibiting the JAK2/STAT3 signaling pathway,thereby reducing oxidative stress and RGC apoptosis.

Centrosome amplification is a hallmark of malignant tumours and has been established as a critical carcinogenic factor driving the progression of various malignancies.This review systematically outlines the structural com-position and replication cycle of the centrosome,thoroughly analyses the specific manifestations of abnormal centrosome amplification in tumor cells,and explores the molecular mechanisms underlying centrosome amplification.Furthermore,this review highlights therapeutic strategies targeting abnormal centrosome expansion and their potential application in tu-mor intervention.In-depth investigations of centrosome amplification mechanisms and intervention pathways may offer novel insights for early cancer diagnosis and precision therapy.

Purpose To evaluate the value of preoperative MRI-based radiomic models for assessing tumor-infiltrating CD8+T-cell expression in glioblastoma patients,and to identify the most stable and efficient radiomic feature region for predicting prognosis following immunotherapy.Materials and Methods This retrospective study included 150 patients with histopathologically confirmed glioblastoma from Lanzhou University Second Hospital(January 2018 to April 2022).Tumor-infiltrating CD8+T-cell expression was quantitatively assessed using immunohistochemical staining,with patients stratified into CD8-high and CD8-low expression groups based on overall survival.A total of 1 185 radiomic features were extracted from each patient's contrast-enhanced T1C and T2WI images,covering the original tumor region and sequentially expanded peritumoral regions(2.5 mm,5.0 mm,7.5 mm,10.0 mm,12.5 mm,15.0 mm morphological dilation of tumor core+peritumoral area).Feature selection was performed using variance threshold,minimum redundancy maximum relevance,and least absolute shrinkage and selection operator methods.XGBoost classifier was employed to construct clinical,radiomic,and clinical-radiomic multimodal combined prediction models.Diagnostic performance was evaluated using receiver operating characteristic curve analysis.Results The radiomic model based on tumor expansion of 7.5 mm(tumor+peritumoral region)demonstrated optimal predictive performance.The clinical-radiomic multimodal combined model showed superior predictive capability compared to clinical and radiomic models alone,achieving an area under the curve of 0.991 and accuracy of 99.0%in the training set,and area under the curve of 0.840 with accuracy of 80.0%in the validation set.Conclusion MRI radiomics provides a feasible approach for evaluating tumor-infiltrating CD8+T-cell expression in glioblastoma patients,offering potential for preoperative prognosis prediction.

Objective To explore the impact of peptidylarginine deiminase 2(Padi2)-knockout on depressive-like behaviors in socially isolated mice.Methods Using CRISPR/Cas9 technology,a Padi2-knockout(Padi2-/-)mouse model with a C57BL/6J background was established,and the effect of Padi2 knockout was identified by genotyping and RT-qPCR detection.Six-week-old male Padi2-/- mice and wild-type C57BL/6J mice were selected and divided into normal rearing and social isolation groups,with 15 mice per group.The normal rearing group mice were housed with 5 mice per cage,and the social isolation group was housed with 1 mouse per cage,and weighed once a week.After 4 weeks,forced swimming and open field tests were conducted.After the behavioral experiments,brain tissues were taken from mice in each group,and changes in microglia in the brains were detected by immunofluorescence.Results We successfully established Padi2-/- mice.There was no difference in behavior between Padi2-/- mice and C57BL/6J mice in the normal rearing group.After social isolation,compared with C57BL/6J mice,Padi2-/- mice showed a significant increase in depressive symptoms,obvious weight gain,and a significant increase in the number of microglia in brain tissue.Conclusions Padi2 knockout exacerbated depressive-like behaviors and obesity in socially isolated mice,indicating that Padi2 is involved in the progression of depression and may be an effective target for the prevention and treatment of depression.

OBJECTIVE To explore the expression levels of microribonucleic acid-21(miR-21)and helper T cell(Th)1/Th2 cytokines in peripheral blood of the asthma children complicated with respiratory tract infections of vi-ruses and analyze the significance.METHODS A total of 90 asthma children with respiratory tract infections of vi-ruses(the infection group)and 43 asthma children without respiratory tract infections of viruses(the no infection group)who were treated in the 904th Hospital of the Joint Logistics Support Force of The People's Liberation Ar-my of China were enrolled in the study,and the clinical data were collected from the enrolled patients.The distri-bution of viruses from the children of the infection group was analyzed.The levels of peripheral blood miR-21,Th1 cytokines[interferon-γ(IFN-γ),interleukin(IL)-2]and Th2 cytokines[IL-4,IL-5]were compared between the two groups.The children of the infection group were divided into the mild group,the moderate group and the severe group according to the severity of disease.The levels of the above indexes were compared among the chil-dren with the various degree of illness condition.The association of the illness condition with miR-21,IFN-γ,IL-2,IL-4 and IL-5 was analyzed.RESULTS Respiratory syncytial virus was dominant among the respiratory tract vi-ruses in the infection group,accounting for 32.22％.The levels of IFN-γ and IL-2 of the infection group were low-er than those of the no infection group(P＜0.05),the levels of miR-21,IL-4 and IL-5 of the infection group were higher than those of the no infection group(P＜0.05).The levels of IFN-γ and IL-2 of the severe group were lower than those of the mild group and the moderate group(P＜0.05),while the levels of miR-21,IL-4 and IL-5 of the severe group were higher than those of the mild group and the moderate group(P＜0.05);the levels of IFN-γ and IL-2 of the moderate group were lower than those of the mild group(P＜0.05),and the levels of miR-21,IL-4 andIL-5 of the moderate group were higher than those of the mild group(P＜0.05).Spearman correlation analysis showed that the illness condition of the children was positively correlated with miR-21,IL-4 and IL-5(P＜0.05),which was negatively correlated with IFN-γ and IL-2(P＜0.05).CONCLUSIONS The respiratory tract infections of viruses may aggravate the abnormal rise of peripheral blood miR-21 and the imbalance of Th1/Th2.The above indexes are closely associated with the illness condition of the children.

Objective To investigate the imaging features of post-transplant lymphoproliferative disorder(PTLD)after allogeneic hematopoietic stem cell transplantation(allo-HSCT).Methods The imaging findings of EB virus-associated PTLD(EBV-PTLD)after allo-HSCT in 8 patients with hematological diseases were retrospectively analyzed,including 1 case with acute lymphoblastic leukemia(ALL),3 cases with acute myeloid leukemia(AML),1 case with myelodysplastic syndrome(MDS)and 3 cases with aplas-tic anemia(AA).All patients were positive for EB virus(EBV)after transplantation.MRI,CT plain scan and enhanced scan were performed to analyze the imaging features of all lesions.Results Central nervous system(CNS)was involved in 5 cases,pharyngeal lymphatic ring and left lacrimal sac was involved in 1 case,liver was involved in 1 case,intraperitoneal and retroperitoneal lymph nodes was involved in 1 case.All patients were pathologically confirmed with PTLD.Conclusion PTLD may involve multiple organs and tissues after hematopoietic stem cell transplantation with different onset time.Timely MRI and CT examination,combined with PTLD screening and EBV detection,can detect and diagnose PTLD lesions in early stage,providing important clinical value.

Stroke imposes a tremendous burden on patients'families and society due to its high rates of mortality,disability,and recurrence.Advances in neuroimaging technologies have provided critical theoretical foundations for investigating the pathophysiological mechanisms of stroke,as well as enabling early clinical intervention and personalized rehabilitation.This article reviewed the application of five commonly used magnetic resonance imaging(MRI)techniques in acupuncture therapy for stroke,including functional MRI(fMRI)for cerebral blood oxygen metabolism,magnetic resonance spectroscopy(MRS),diffusion MRI(dMRI),perfusion MRI(pMRI),and structural MRI(sMRI).By examining functional,metabolic,structural,and hemodynamic aspects,these imaging modalities offer evidence to validate the multi-target effect and efficacy of acupuncture in stroke treatment.