ObjectiveTo investigate the immunomodulatory effect of polysaccharides from Astragali Radix-Angelicae Sinensis Radix herb pair（Qi-gui polysaccharides） on lipopolysaccharide（LPS）-induced RAW264.7 macrophages and to characterize the structure of the active component Qi-gui homogeneous polysaccharide（AAPS-4a）， and evaluate its protective effect on ulcerative colitis（UC）. MethodsThe effects of six Qi-gui polysaccharides（0.01-100 mg·L^-1） on the proliferation of RAW264.7 cells were assessed by cell proliferation and activity assay（CCK-8）， and enzyme-linked immunosorbent assay（ELISA） was used to investigate the effects of the six polysaccharides（3， 10 mg·L^-1） on the secretion levels of tumor necrosis factor（TNF）-α， interferon（IFN）-β， and nitric oxide（NO） in LPS-induced RAW264.7 cells. After screening for active polysaccharides， high-performance size-exclusion chromatography（HPSEC） was used to determine its homogeneity and relative molecular weight， then its characteristic functional groups were identified by Fourier transform infrared spectroscopy（FT-IR）， monosaccharide composition was analyzed by high performance liquid chromatography（HPLC）， methylation analysis combined with gas chromatography-mass spectrometry（GC-MS） was performed to determine the types and linkage modes of sugar residues， and one- and two-dimensional nuclear magnetic resonance（NMR） were used to identify the sugar residue composition and configuration of the active polysaccharide. Finally， experimental animals were divided into the normal group， model group， AAPS-4a low-dose group（50 mg·kg^-1）， AAPS-4a high-dose group（100 mg·kg^-1）， and sulfasalazine（SASP） group （75 mg·kg^-1）. Except for the normal group， the acute UC mouse model was induced using 3.5% dextran sulfate sodium salt（DSS）. Each treatment group was administered the corresponding dose via oral gavage for 7 days， and changes in body weight were recorded. After treatment， the spleen index and disease activity index（DAI） score were calculated， TNF-α and interleukin-6（IL-6） levels in the serum were detected by ELISA， and histopathological changes in colon tissue were observed by hematoxylin-eosin（HE） staining. ResultsAt the cellular level， AAPS-4a exhibited a dose-dependent inhibition of LPS-induced increases in TNF-α， IFN-β， and NO levels（P<0.01）. Structural characterization of AAPS-4a revealed that it was a homogeneous polysaccharide with a relative molecular weight of 7.6×10³ Da， consisting of mannose（Man）， glucose（Glc）， and galactose（Gal） in a molar ratio of 1.3∶23.9∶1.0. It was primarily composed of five sugar residues of 1，6-α-D-Glcp， T-α-D-Glcp， 1，3-β-D-Galp， 1，4-α-D-Manp， and 1，2-α-D-Galp. In vivo experiments showed that compared with the normal group， the model group demonstrated markedly increased DAI score and spleen index， significantly reduced colon length， and significantly elevated levels of TNF-α and IL-6（P<0.01）. Compared with the model group， the AAPS-4a high-dose group significantly reduced the DAI score and spleen index， as well as TNF-α and IL-6 levels， and improved colonic atrophy（P<0.05， P<0.01）. Pathological observations showed that AAPS-4a significantly inhibited inflammatory cell infiltration in colon tissue and alleviated pathological damage. ConclusionAAPS-4a， a neutral homogeneous polysaccharide composed of 1，6-α-D-Glcp， T-α-D-Glcp， 1，3-β-D-Galp， 1，4-α-D-Manp and 1，2-α-D-Galp， is identified as a key bioactive component contributing to the anti-UC effect of the Qi-gui herb pair. Its immunoregulatory and anti-UC properties suggest its potential as a therapeutic agent for UC.

OBJECTIVES: To investigate the regulatory role of nucleotide-bound oligomerized domain-like receptor containing pyrin-domain protein 6 (NLRP6) in liver lipid metabolism and non-alcoholic fatty liver disease (NAFLD). METHODS: Mouse models with high-fat diet (HFD) feeding for 16 weeks (n=6) or with methionine choline-deficient diet (MCD) feeding for 8 weeks (n=6) were examined for the development of NAFLD using HE and oil red O staining, and hepatic expressions of NLRP6 were detected with RT-qPCR, Western blotting, and immunohistochemical staining. Cultured human hepatocytes (LO2 cells) with adenovirus-mediated NLRP6 overexpression or knock-down were treated with palmitic acid (PA) in the presence or absence of compound C (an AMPK inhibitor), and the changes in cellular lipid metabolism were examined by measuring triglyceride, ATP and β-hydroxybutyrate levels and using oil red staining, RT-qPCR, and Western blotting. RESULTS: HFD and MCD feeding both resulted in the development of NAFLD in mice, which showed significantly decreased NLRP6 expression in the liver. In PA-treated LO2 cells, NLRP6 overexpression significantly decreased cellular TG content and lipid deposition, while NLRP6 knockdown caused the opposite effects. NLRP6 overexpression in PA-treated LO2 cells also increased mRNA and protein expressions of PGC1A and CPT1A, levels of ATP and β-hydroxybutyrate, and the phosphorylation level of AMPK pathway; the oxidative decomposition of lipids induced by Ad-NLRP6 was inhibited by the use of AMPK inhibitors. CONCLUSIONS NLRP6 overexpression promotes lipid oxidation and decomposition through AMPK/CPT1A/PGC1A to alleviate lipid deposition in hepatocytes.
Non-alcoholic Fatty Liver Disease/metabolism* ; Animals ; Hepatocytes/metabolism* ; Lipid Metabolism ; Mice ; Humans ; Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha ; AMP-Activated Protein Kinases/metabolism* ; Carnitine O-Palmitoyltransferase/metabolism* ; Diet, High-Fat ; Male ; Mice, Inbred C57BL ; Signal Transduction

Acute-on-chronic liver failure （ACLF） refers to severe liver dysfunction that occurs on the basis of chronic liver diseases， and it is characterized by rapid disease progression， poor prognosis， and high mortality rate. In recent years， inflammatory markers have become a research hotspot due to their significant role in assessing the prognosis of ACLF. This article reviews the advances in the application of inflammatory markers in assessing the prognosis of ACLF， such as systemic immune inflammatory index， lymphocyte-white blood cell ratio， neutrophil-lymphocyte ratio， and C-reactive protein， and discusses their clinical value and future research directions， in order to provide a theoretical basis for the early intervention and prognosis management of ACLF patients.

OBJECTIVE To investigate the expression and clinical significance of Periostin in tissues of patients with chronic rhinosinusitis(CRS).METHODS Real-time quantitative PCR and immunohistochemistry were used to detect periostin expression in eosinophilic CRS with nasal polyps(ECRSwNP),non-eosinophilic CRS with nasal polyps(non-ECRSwNP),CRS without nasal polyps(CRSsNP),and control tissues.Correlations between periostin levels and blood eosinophil percentage(Eos%),Lund-Mackay score,modified endoscopic score,and Japanese epidemiological survey of refractory eosinophilic chronic rhinosinusitis(JESREC)score were analyzed.Additionally,changes in SNOT-22 and VAS scores were compared at different preoperative and postoperative times.The predictive value of periostin for ECRSwNP was evaluated using receiver operating characteristic(ROC)curve analysis.RESULTS Periostin expression was detected in all groups(ECRSwNP,non-ECRSwNP,CRSsNP,and controls),with predominant localization in the basement membrane and mucosal subepithelial lamina propria.Significantly elevated periostin levels were detected in the ECRSwNP group compared to the other three groups(P<0.001).Furthermore,Periostin mRNA expression showed significant positive correlations with blood Eos%,JESREC score,and Lund-Mackay score.SNOT-22 and VAS scores were significantly elevated in the ECRSwNP group at preoperative evaluation and 9 months postoperatively(P<0.001).ROC curve analysis demonstrated that periostin had a substantial predictive value for ECRSwNP(AUC=0.957).CONCLUSION Periostin plays a crucial role in the pathogenesis of chronic rhinosinusitis,contributing to the diagnosis,severity assessment,and prognosis evaluation of ECRSwNP,while offering potential therapeutic targets for CRS management.

Objective To compare the clinical efficacy and short-term prognosis of laparoscopic radical resection of right colon cancer guided by superior mesenteric artery and superior mesenteric vein.Methods 80 patients with right colon cancer of cT2-4 and/or N0-2M0 admitted from January 2020 to October 2022 were selected as the research objects,and they were randomly divided into observation group and control group,with 40 patients in each group.The observation group was treated with SMA-oriented laparoscopic radical resection of right colon cancer,while the control group was treated with SMV-oriented laparoscopic radical resection of right colon cancer.The curative effect and prognosis of the two groups were compared.Results There was no significant difference between the two groups in general condition,operation time,gastric tube placement time,recovery time of farting,postoperative fasting time,postoperative drainage time,postoperative nutritional index,total incidence of complications and postoperative hospitalization time(P>0.05).The lymph nodes in the observation group were significantly more than those in the control group,and the difference was statistically significant(P<0.05).In the observation group,the lymph nodes in the anterior and left side of superior mesenteric artery were examined(No.D3),and 273 lymph nodes were detected,and Seven patients(17.5% )were diagnosed with D3 metastasis,and 13 lymph nodes were positive(5.2% ).Conclusion Laparoscopic radical resection of right colon cancer guided by superior mesenteric artery,without increasing the incidence of complications and high safety,can more thoroughly clean lymph nodes and reduce tumor recurrence,which is expected to significantly improve the prognosis of patients.

Objective:To explore the risk factors for stone recurrence after laparoscopic common bile duct exploration (LCBDE) combined with laparoscopic cholecystectomy (LC) and to develop a risk prediction model.Methods:Clinical data of 344 patients with bile duct stones who underwent LCBDE combined with LC at Hebei General Hospital from January 2016 to March 2022 were retrospectively analyzed, including 165 males and 179 females, aged (62.72±13.56) years old. Patients were divided into two groups based on whether stones recurred during the follow-up period: recurrence group ( n=37) and non-recurrence group ( n=307). Clinical data such as common bile duct diameter, stone size, number of stones and duration of T-tube drainage were collected from the patients. Logistic regression was used to analyze the risk factors for postoperative stone recurrence, and then developed a logistic regression model. The predictive efficacy of the model was assessed by the area under the receiver operating characteristic (ROC) curve, and the Hosmer-Lemeshow test. Results:The results of multifactorial logistic regression analysis showed that patients with ≥2 choledochal stones had a high risk of stone recurrence after LCBDE combined with LC ( OR=3.094, 95% CI: 1.069-8.954, P=0.037). In contrast, regular postoperative oral choleretic medication was a protective factor for stone recurrence after LCBDE combined with LC ( OR=0.160, 95% CI: 0.072-0.354, P=0.001). A logistic regression model, based on the number of common bile duct stones and regular postoperative oral choleretic medication, was developed to predict the recurrence of bile duct stones in patients who underwent LCBDE combined with LC. The area under the ROC curve for this model was found to be 0.821 (95% CI: 0.758-0.885). The Hosmer-Lemeshow test, χ 2=7.26, P=0.509, suggested that there is good agreement between the model's predicted probabilities and ideal probabilities. Conclusions:The number of stones (≥2) is an independent risk factor for stone recurrence after LCBDE combined with LC in patients with bile duct stones. Regular postoperative oral choleretic medication is a protective factor for stone recurrence after LCBDE combined with LC. Predictive models based on the number of choledochal stones and regular postoperative oral choleretic medication have better efficacy in predicting postoperative stone recurrence.

Objective Atrial fibrillation(AF)is a disease of high heterogeneity,and the association between AF phenotypes and the outcome of different catheter ablation strategies remains unclear.Conventional classification of AF(e.g.according to duration,atrial size,and thromboembolism risk)fails to provide reference for the optimal stratification of the prognostic risks or to guide individualized treatment plan.In recent years,research on machine learning has found that cluster analysis,an unsupervised data-driven approach,can uncover the intrinsic structure of data and identify clusters of patients with pathophysiological similarity.It has been demonstrated that cluster analysis helps improve the characterization of AF phenotypes and provide valuable prognostic information.In our cohort of AF inpatients undergoing radiofrequency catheter ablation,we used unsupervised cluster analysis to identify patient subgroups,to compare them with previous studies,and to evaluate their association with different suitable ablation patterns and outcomes.Methods The participants were AF patients undergoing radiofrequency catheter ablation at West China Hospital between October 2015 and December 2017.All participants were aged 18 years or older.They underwent radiofrequency catheter ablation during their hospitalization.They completed the follow-up process under explicit informed consent.Patients with AF of a reversible cause,severe mitral stenosis or prosthetic heart valve,congenital heart disease,new-onset acute coronary syndrome within three months prior to the surgery,or a life expectancy less than 12 months were excluded according to the exclusion criteria.The cohort consisted of 1 102 participants with paroxysmal or persistent/long-standing persistent AF.Data on 59 variables representing demographics,AF type,comorbidities,therapeutic history,vital signs,electrocardiographic and echocardiographic findings,and laboratory findings were collected.Overall,data for the variables were rarely missing(<5% ),and multiple imputation was used for correction of missing data.Follow-up surveys were conducted through outpatient clinic visits or by telephone.Patients were scheduled for follow-up with 12-lead resting electrocardiography and 24-hours Holter monitoring at 3 months and 6 months after the ablation procedure.Early ablation success was defined as the absence of documented AF,atrial flutter,or atrial tachycardia>30 seconds at 6-month follow-up.Hierarchical clustering was performed on the 59 baseline variables.All characteristic variables were standardized to have a mean of zero and a standard deviation of one.Initially,each patient was regarded as a separate cluster,and the distance between these clusters was calculated.Then,the Ward minimum variance method of clustering was used to merge the pair of clusters with the minimum total variance.This process continued until all patients formed one whole cluster.The"NbClust"package in R software,capable of calculating various statistical indices,including pseudo t2 index,cubic clustering criterion,silhouette index etc,was applied to determine the optimal number of clusters.The most frequently chosen number of clusters by these indices was selected.A heatmap was generated to illustrate the clinical features of clusters,while a tree diagram was used to depict the clustering process and the heterogeneity among clusters.Ablation strategies were compared within each cluster regarding ablation efficacy.Results Five statistically driven clusters were identified:1)the younger age cluster(n=404),characterized by the lowest prevalence of cardiovascular and cerebrovascular comorbidities but the highest prevalence of obstructive sleep apnea syndrome(14.4% );2)a cluster of elderly adults with chronic diseases(n=438),the largest cluster,showing relatively higher rates of hypertension,diabetes,stroke,and chronic obstructive pulmonary disease;3)a cluster with high prevalence of sinus node dysfunction(n=160),with patients showing the highest prevalence of sick sinus syndrome and pacemaker implantation;4)the heart failure cluster(n=80),with the highest prevalence of heart failure(58.8% )and persistent/long-standing persistent AF(73.7% );5)prior coronary artery revascularization cluster(n=20),with patients of the most advanced age(median:69.0 years old)and predominantly male patients,all of whom had prior myocardial infarction and coronary artery revascularization.Patients in cluster 2 achieved higher early ablation success with pulmonary veins isolation alone compared to extensive ablation strategies(79.6% vs.66.5%;odds ratio[OR]=1.97,95% confidence interval[CI]:1.28-3.03).Although extensive ablation strategies had a slightly higher success rate in the heart failure group,the difference was not statistically significant.Conclusions This study provided a unique classification of AF patients undergoing catheter ablation by cluster analysis.Age,chronic disease,sinus node dysfunction,heart failure and history of coronary artery revascularization contributed to the formation of the five clinically relevant subtypes.These subtypes showed differences in ablation success rates,highlighting the potential of cluster analysis in guiding individualized risk stratification and treatment decisions for AF patients.

Objective:To investigate the diagnostic accuracy of serological indicators and evaluate the diagnostic value of a new established combined serological model on identifying the minimal hepatic encephalopathy (MHE) in patients with compensated cirrhosis.Methods:This prospective multicenter study enrolled 263 compensated cirrhotic patients from 23 hospitals in 15 provinces, autonomous regions and municipalities of China between October 2021 and August 2022. Clinical data and laboratory test results were collected, and the model for end-stage liver disease (MELD) score was calculated. Ammonia level was corrected to the upper limit of normal (AMM-ULN) by the baseline blood ammonia measurements/upper limit of the normal reference value. MHE was diagnosed by combined abnormal number connection test-A and abnormal digit symbol test as suggested by Guidelines on the management of hepatic encephalopathy in cirrhosis. The patients were randomly divided (7∶3) into training set ( n=185) and validation set ( n=78) based on caret package of R language. Logistic regression was used to establish a combined model of MHE diagnosis. The diagnostic performance was evaluated by the area under the curve (AUC) of receiver operating characteristic curve, Hosmer-Lemeshow test and calibration curve. The internal verification was carried out by the Bootstrap method ( n=200). AUC comparisons were achieved using the Delong test. Results:In the training set, prevalence of MHE was 37.8% (70/185). There were statistically significant differences in AMM-ULN, albumin, platelet, alkaline phosphatase, international normalized ratio, MELD score and education between non-MHE group and MHE group (all P<0.05). Multivariate Logistic regression analysis showed that AMM-ULN [odds ratio ( OR)=1.78, 95% confidence interval ( CI) 1.05-3.14, P=0.038] and MELD score ( OR=1.11, 95% CI 1.04-1.20, P=0.002) were independent risk factors for MHE, and the AUC for predicting MHE were 0.663, 0.625, respectively. Compared with the use of blood AMM-ULN and MELD score alone, the AUC of the combined model of AMM-ULN, MELD score and education exhibited better predictive performance in determining the presence of MHE was 0.755, the specificity and sensitivity was 85.2% and 55.7%, respectively. Hosmer-Lemeshow test and calibration curve showed that the model had good calibration ( P=0.733). The AUC for internal validation of the combined model for diagnosing MHE was 0.752. In the validation set, the AUC of the combined model for diagnosing MHE was 0.794, and Hosmer-Lemeshow test showed good calibration ( P=0.841). Conclusion:Use of the combined model including AMM-ULN, MELD score and education could improve the predictive efficiency of MHE among patients with compensated cirrhosis.

[摘 要] 目的：探讨吴茱萸碱（Evo）是否通过调控lncRNA LINC00858表达调控神经母细胞瘤SK-N-SH细胞的增殖、迁移及侵袭。方法：在体外以3、6、12 μmol/L Evo处理人神经母细胞瘤SK-N-SH细胞，利用RNA干扰技术分别将si-NC、si-LINC00858转染至SK-N-SH细胞，将pcDNA、pcDNA-LINC00858转染至SK-N-SH细胞并经12 μmol/L Evo处理，实验分为对照组、Evo低剂量组、Evo中剂量组、Evo高剂量组、si-NC组、si-LINC00858组、Evo+pcDNA组、Evo+pcDNA-LINC00858组。采用qPCR法检测各组细胞LINC00858的表达量，MTT、Transwell实验分别检测细胞的增殖、迁移、侵袭能力，WB法检测细胞中cyclinD1、MMP-2、MMP-9和p21蛋白的表达。结果：与对照组相比，Evo低、中、高剂量组SK-N-SH细胞中LINC00858表达均显著降低（均P<0.05），细胞增殖抑制率显著升高、迁移及侵袭细胞数显著减少（均P<0.01），cyclinD1、MMP-2、MMP-9蛋白表达降低、p21蛋白表达升高（均P<0.01）。与si-NC组相比，si-LINC00858组细胞的增殖抑制率、迁移和侵袭细胞数及相关蛋白表达变化同Evo低、中、高剂量组。与Evo+pcDNA组相比，Evo+pcDNA-LINC00858组细胞的增殖抑制率显著降低、迁移及侵袭细胞数均显著增多（均P<0.01），cyclinD1、MMP-2、MMP-9蛋白表达升高、p21蛋白表达降低（均P<0.05）。结论：Evo通过下调LINC00858表达抑制神经母细胞瘤SK-N-SH细胞的增殖、迁移及侵袭。