[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.

ObjectiveTo investigate the value of preoperative dual-layer spectral detector CT （DLCT） parameters in predicting lymph node metastasis （LNM） of non-functional pancreatic neuroendocrine neoplasms （NF-pNENs）. MethodsA retrospective study was conducted on patients with pathologically confirmed NF-pNENs who underwent preoperative contrast-enhanced DLCT examination between September 2019 and July 2024. Regions of interest （ROIs） were manually delineated on three representative slices of the solid component of the lesion， and the mean values were recorded. Conventional CT parameters， monochromatic energy images （40 keV， 70 keV， 100 keV）， iodine concentration maps， and effective atomic number （Z_eff） maps in the pancreatic parenchymal phase and portal venous phase were acquired. A total of 34 parameters， including clinical characteristics， imaging features， conventional CT and DLCT quantitative parameters， were analyzed. Intergroup differences were compared between the LNM and non-LNM groups. Independent predictive factors were screened using logistic regression and least absolute shrinkage and selection operator （LASSO） regression. Receiver operating characteristic （ROC） curves were generated for each predictive model， and the area under the curve （AUC） was calculated to evaluate the predictive efficacy. ResultsA total of 82 patients with NF-pNENs were enrolled， including 37 males and 45 females， with a mean age of 49.7±14.7 years. Thirty patients were performed with LNM and 52 were with non-LNM. After univariate logistic regression， LASSO regression and multivariate stepwise logistic regression analyses， body mass index （BMI）， organ invasion， normalized CT value in the portal venous phase （nCTv） and normalized iodine concentration in the portal venous phase （nICv） were identified as independent predictive factors. The AUCs of the clinical-radiological qualitative model （BMI+organ invasion）， conventional CT model （nCTv）， DLCT model （nICv） and comprehensive model （organ invasion+nICv） were 0.815 （0.719， 0.910）， 0.750 （0.642， 0.858）， 0.829 （0.733， 0.925） and 0.865 （0.879， 0.953）， respectively. The comprehensive model combining clinical-radiological and DLCT characteristics exhibited superior predictive performance. ConclusionDLCT parameters are more effective than conventional CT parameters in predicting LNM in NF-pNENs， enabling non-invasive preoperative prediction and assisting clinical decision-making.

ObjectiveTo investigate the protective effect of ellagic acid （EA） against bleomycin （BLM）-induced pulmonary fibrosis and associated pulmonary function impairment in mice， and to elucidate its relationship with the regulation of the interleukin（IL）-17/ nuclear factor （NF）-κB/ matrix metalloproteinase （MMP）9 signaling axis. MethodsA pulmonary fibrosis model was established in male C57BL/6J mice via intratracheal BLM injection. Interventions included IL-17 neutralizing antibody， Ixekizumab， SB-3CT， PDTC， or EA were used to modulate the IL-17/NF-κB/MMP9 signaling axis. Pathological changes in lung tissue were observed via HE， Masson， and Sirius red staining. Pulmonary function was assessed using a pulmonary function test （PFT）. Western blot and qRT-PCR were employed to detect related protein and gene expression. Network pharmacology was utilized to predict the potential targets of EA. A protein-protein interaction （PPI） network was constructed using the STRING database and Cytoscape. Molecular docking was performed to validate the binding capability of EA to core targets. ResultsBLM successfully induced obvious pulmonary fibrosis and lung dysfunction in mice， significantly elevating the level of the pro-inflammatory cytokine IL-17. This was associated with the activation of the transcription factor NF-κB p65， leading to the upregulation of the pro-fibrotic factor MMP9. Inhibition of the IL-17/NF-κB/MMP9 signaling axis markedly alleviated the degree of pulmonary fibrosis. EA intervention significantly suppressed the BLM-induced increase in IL-17， blocked the activation of the NF-κB/MMP9 pathway， and consequently reduced lung fibrotic lesions and improved pulmonary function. ConclusionEA may effectively ameliorate BLM-induced pulmonary fibrosis and lung dysfunction in mice， likely by inhibiting the inflammatory and fibrotic responses mediated through the IL-17/NF-κB/MMP9 signaling pathway.

ObjectiveTo investigate the value of preoperative dual-layer spectral detector CT （DLCT） parameters in predicting lymph node metastasis （LNM） of non-functional pancreatic neuroendocrine neoplasms （NF-pNENs）. MethodsA retrospective study was conducted on patients with pathologically confirmed NF-pNENs who underwent preoperative contrast-enhanced DLCT examination between September 2019 and July 2024. Regions of interest （ROIs） were manually delineated on three representative slices of the solid component of the lesion， and the mean values were recorded. Conventional CT parameters， monochromatic energy images （40 keV， 70 keV， 100 keV）， iodine concentration maps， and effective atomic number （Z_eff） maps in the pancreatic parenchymal phase and portal venous phase were acquired. A total of 34 parameters， including clinical characteristics， imaging features， conventional CT and DLCT quantitative parameters， were analyzed. Intergroup differences were compared between the LNM and non-LNM groups. Independent predictive factors were screened using logistic regression and least absolute shrinkage and selection operator （LASSO） regression. Receiver operating characteristic （ROC） curves were generated for each predictive model， and the area under the curve （AUC） was calculated to evaluate the predictive efficacy. ResultsA total of 82 patients with NF-pNENs were enrolled， including 37 males and 45 females， with a mean age of 49.7±14.7 years. Thirty patients were performed with LNM and 52 were with non-LNM. After univariate logistic regression， LASSO regression and multivariate stepwise logistic regression analyses， body mass index （BMI）， organ invasion， normalized CT value in the portal venous phase （nCTv） and normalized iodine concentration in the portal venous phase （nICv） were identified as independent predictive factors. The AUCs of the clinical-radiological qualitative model （BMI+organ invasion）， conventional CT model （nCTv）， DLCT model （nICv） and comprehensive model （organ invasion+nICv） were 0.815 （0.719， 0.910）， 0.750 （0.642， 0.858）， 0.829 （0.733， 0.925） and 0.865 （0.879， 0.953）， respectively. The comprehensive model combining clinical-radiological and DLCT characteristics exhibited superior predictive performance. ConclusionDLCT parameters are more effective than conventional CT parameters in predicting LNM in NF-pNENs， enabling non-invasive preoperative prediction and assisting clinical decision-making.

ObjectiveTo investigate the effect of trophoblast-derived mesenchymal stem cell （T-MSC） on concanavalin A （Con A）-induced immune-mediated liver injury in mice， as well as its association with the changes in inflammatory response. MethodsMorphological assessment， flow cytometry， and tri-lineage differentiation assay were used to observe the biological characteristics of T-MSC. In the pilot experiment， mice were divided into PBS group， T-MSC monotherapy group， and Con A groups at a concentration of 10， 15， or 20 mg/kg to evaluate the safety of T-MSC monotherapy on liver function and determine the appropriate dose for modeling. In the formal experiment， mice were divided into healthy control group （HC group）， Con A group， and Con A+T-MSC group， with 5 mice in each group. The mice were given injection of Con A （15 mg/kg） via the caudal vein to establish a mouse model of immune-mediated liver injury， while the mice in the T-MSC monotherapy group were given transfusion of 1×10⁶ T-MSCs via the caudal vein at 3 hours after modeling， and samples were collected 12 hours later. The serum levels of alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， and lactate dehydrogenase （LDH） were measured； liver histopathological changes were observed， and Ishak inflammatory activity score and inflammatory necrosis area were assessed； the serum levels of inflammatory cytokines were measured； transcriptome sequencing， immunofluorescence assay， and Western blot were used to measure the changes in inflammation-related molecules in liver tissue. The Shapiro-Wilk test was used to assess the normality of continuous data， and the homogeneity of variance test was also performed； a one-way analysis of variance was used for comparison of normally distributed data with homogeneity of variance between multiple groups， and the Tukey’s post-hoc test was used for further comparison between two groups； the Kruskal-Wallis H test was used for comparison of non-normally distributed data with heterogeneity of variance between multiple groups， and the Wilcoxon test was used for further comparison between two groups. ResultsT-MSC had a long spindle shape， showed the characteristics of fibroblast-like morphology and adherent growth， and possessed the osteogenic， chondrogenic， and adipogenic differentiation abilities. Flow cytometry showed high expression levels of CD90， CD105， and CD73 in T-MSC， with a positive rate of 96.8%， 99.0%， and 99.9%， respectively， while there was low or no expression of CD31， human leukocyte antigen DR， CD45， and CD34， with a positive rate of 0.120%， 0， 0.061%， and 0， respectively. Administration of T-MSC alone did not cause obvious abnormalities in ALT， AST， or LDH. Con A at the concentrations of 10， 15， and 20 mg/kg all increased the serum levels of biochemical parameters for liver injury in a dose-dependent manner， with mortality observed in the 20 mg/kg group. In the formal experiment， compared with the HC group， the Con A group had significant increases in the serum levels of ALT， AST， and LDH （P<0.01）， and after T-MSC treatment， there were significant reductions in these markers compared with the Con A group （P<0.01）. HE staining showed that the Con A group had disordered structure of the liver， accompanied by marked inflammatory cell infiltration and focal/patchy necrosis， and these pathological changes were alleviated after T-MSC treatment. Compared with the HC group， the Con A group had significant increases in Ishak score and the percentage of inflammatory necrosis area （P<0.001）， while there were significant reductions in these indicators after T-MSC treatment compared with the Con A group （P<0.01）. ELISA showed that compared with the HC group， the Con A group had significant increases in the serum levels of interleukin-21 （IL-21）， interleukin-1β （IL-1β）， interleukin-2 （IL-2）， interleukin-17A （IL-17A）， and tumor necrosis factor-α （TNF-α） （P<0.05）， and there were significant reductions in these markers after T-MSC treatment compared with the Con A group （P<0.05）， with no significant differences in IL-21， IL-1β， IL-17A， and TNF-α compared with the HC group （P>0.05）； although there was a significant reduction in the level of IL-2， the Con A+T-MSC group still had a significantly higher level of IL-2 than the HC group （P=0.001）. In contrast， compared with the HC group， the Con A group had a significant increase in IL-10 （P<0.05）， with a further increase after T-MSC treatment （P<0.05）. The transcriptome analysis showed that T-MSC treatment downregulated the expression of the inflammation- and chemokine-related genes including Ccl2， Cxcl10， Il-21， and Il-6 and inhibited the inflammation-related pathways such as TNF， IL-17， nuclear factor-κB， and JAK-STAT. Immunofluorescence assay showed that compared with the HC group， the Con A group had a significant increase in the expression of IL-21 in the liver （P<0.001）， and there was a significant reduction in IL-21 after T-MSC treatment compared with the Con A group （P<0.001）； the results of Western blot showed the same trend. ConclusionT-MSC can alleviate Con A-induced immune-mediated liver injury in mice， as evidenced by the improvement in liver biochemical parameters， the alleviation of histopathological damage， and the overall downregulation of inflammation-related molecules. T-MSC exerts marked anti-inflammatory and immunomodulatory effects in this model， and IL-21 shows a significant change in inflammatory response and can be used as a candidate molecule for further investigation.

Angle class Ⅱ malocclusion is often characterized by mandibular retraction and lip incompetence,which affects the patient's lateral appearance and may even lead to upper airway stenosis.It can be classified into dental and skeletal types.For skeletal class Ⅱ malocclusion patients with mandibular retraction during the peak growth period,mandibular anterior guidance with a functional orthodontic appliance is generally considered as the optimal clinical treatment approach.At present,there remains a paucity of clinical reports on the clinical application of bracket-free clear aligners in mandibular anterior guidance,both domestically and internationally.This article presented a case of an adolescent patient with skeletal class Ⅱ malocclusion accompanied with deep overbite treated with bracket-free clear aligner for mandibular anterior guidance in combination with intermaxillary class Ⅱ traction.During the treatment,vertical correction involved anterior intrusion of the anterior teeth to improve the deep overbite,while horizontal correction included maxillary and mandibular expansion to coordinate the width of the dental arches,and asymmetric anterior guidance was used to correct the midline deviation.After 35 months of treatment,the patient's convex facial profile and mandibular retrusion were significantly improved.The subspinale-nasion-supramentale angle(ANB)was decreased from 6.8° to 3.9°,the overbite and overjet were normalized,and the bilateral canine and molar reached a neutral relationship.The mentolabial sulcus depth(Si-LiPg′)and the soft tissue thickness of pogonion to pogonion(Pm-Pm′)were decreased,resulting in a shallower mentolabial sulcus and a more harmonized lateral facial soft tissue profile.The mandibular incisor to mandibular plane angle(IMPA)was decreased from 116.6° to 110.7°,indicating retraction of the lower incisors during mandibular anterior guidance.In conclusion,the orthodontic strategy of mandibular advancement with clear aligners in skeletal class Ⅱ malocclusion patients can avoid excessive overcompensation of the upper and lower anterior teeth and shorten the orthodontic treatment cycle.

Objective:To identify the main components of Huanglian Jiedu Decoction(HLJDD)using ultra-high-performance liquid chromatography-quadrupole-time of flight-mass spectrometry(UPLC-Q-TOF-MS),and to explore the potential mechanism of action of HLJDD in the treatment of gouty arthritis(GA)using network pharmacology and molecular docking methods.Methods:We identi-fied the chemical components of HLJDD by combining UPLC-Q-TOF-MS data acquired in both positive and negative ion modes with reference standards,relevant literature,and database searches.We analyzed the potential therapeutic mechanism of HLJDD for GA by using network pharmacology to determine the intersection targets between the active ingredients of HLJDD and GA for further enrich-ment analysis and visual network mapping.The binding affinity of the active ingredients with the intersection targets was validated through molecular docking.Results:A total of 47 components were identified by UPLC-Q-TOF-MS;54 key components of HLJDD for GA treatment and 37 intersection targets were determined by net-work pharmacology;and the top 10 key targets by Degree value were obtained by protein-protein interaction analysis.The Gene On-tology functional enrichment analysis revealed 20 biological pro-cesses,7 cellular components,and 8 molecular functions.The Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis demonstrated 96 GA-related intervention pathways,in which inflammatory signaling pathways such as interleukin-17(IL-17)and tu-mor necrosis factor(TNF)were involved.Molecular docking verified that the key components of HLJDD had high binding affinity with the core targets.Conclusion:The identified key components in HLJDD,such as phellodendrine,coptisine,wogonin,and β-sitosterol,may alleviate GA by regulating multiple core targets in the IL-17 and TNF pathways,such as PTSG2,which provides a theoretical ba-sis for future investigation into the mechanism of action of HLJDD.

Plutonium isotopes(238Pu,239Pu,240Pu and 241Pu)in the environment are important"fingerprint"nuclides in the study of nuclear activity traceability.The content of plutonium isotopes in the environmental metrics is usually very low,and the measurement of these isotopes,especially 238Pu,using mass spectrometry is seriously interfered with by the coexisting 238U.The analysis of several plutonium isotopes in soil usually requires combination of multiple measurement techniques,which leads to a long analysis time and large uncertainty in the isotope ratio.In this work,the hydrous titanium oxide(HTiO)precipitated by the hydrolysis of titanium oxydichloride(TiOCl2)under near-neutral condition was used to preconcentrate plutonium from the soil digestion solution,and the highly efficient decontamination of 238U in the sample was achieved by TK200 resin column chromatography with a decontamination factor of 108.Simulation resuts of density functional theory(DFT)showed that NH3 was considered as a promising reaction gas to eliminate the interference of 238U from 238Pu measurement using mass spectrometry due to the significant discrepancy of the chemical reactivity of U+and Pu+with the reactive gas NH3.Experiments confirmed that by optimizing the flow rates of collision gas(He)and reaction gas(NH3),the interference of 238U could be effectively suppressed,and the decontamination factor of 238U was 104.Combined with chemical separation,the overall decontamination factor of 238U could reach 1012 by using the developed method.By combining chemical separation and tandem quadrupole inductively coupled plasmon-mass spectrometry(ICP-MS/MS)measurement,the simultaneous determination of four ultra-trace plutonium isotopes in soil was realized,and the detection limit of plutonium isotopes was at the femtogram level.Analysis of the international standard reference materials(NIST-SRM-4357 and IAEA-384)showed that the established method could be successfully used for the accurate analysis of ultra-trace four plutonium isotopes(238Pu,239Pu,240Pu and 241Pu)in soil samples.

Poly(lactic-co-glycolide)(PLGA)is a key excipient in long-acting sustained-release preparations,and its degradation properties directly affect the drug release behavior.In this study,PLGA microspheres were prepared by microfluidic techniques,and the morphology changes of the microspheres were observed by scanning electron microscopy(SEM).In alkaline environment,due to the accelerated hydrolysis of ester bonds,the surface of the microspheres was rapidly dissolved and eroded,and the degradation rate was significantly higher than that in acidic environment.High temperature accelerated the degradation of PLGA microspheres.Under neutral and alkaline conditions,the microspheres showed aggregation and adhesion.Under acidic conditions,the microspheres gradually decomposed into irregular fragments.The high ionic strength further promoted the surface corrosion of the microspheres,especially under extreme pH conditions.Simultaneously,PLGA microspheres encapsulating coumarin were prepared to simulate the microsphere formulation.The release rate of coumarin after degradation of the microspheres under different conditions was observed by measuring the absorbance with ultraviolet-visible spectrophotometry.The results were consistent with those of the blank microspheres.This study revealed that the degradation of PLGA microspheres was significantly pH-dependent,temperature sensitive and ion strength responsive.These findings not only helped to understand and optimize the long-term stability and controlled release performance of drug-carrying microspheres,but also provided a theoretical basis for further improvement of PLGA-based drug carrier design.