Metastatic dissemination is the major cause of death from breast-cancer (BC). Fusobacterium nucleatum (F.n) is widely enriched in BC and has recently been identified as one of the high-risk factors for promoting BC metastasis. Here, with an experimental model, we demonstrated that intratumoral F.n induced BC aggressiveness by transcriptionally activating Epithelial-mesenchymal transition-associated genes. Therefore, the F.n may be a potential target to prevent metastasis. Given the fact that cancer-associated fibroblasts (CAFs) are abundant in BC and located near blood vessels, we report an optogenetic system that drives CAF to in situ produce human antibacterial peptide LL37, with the characteristics of biosafety and freely intercellular trafficking, for depleting intratumoral F.n, leading to a 72.1% reduction in lung metastatic nodules number without affecting the balance of the systemic flora. Notably, mild photothermal treatment was found that could normalize CAF, contributing to synergistically inhibiting BC metastasis. In addition, the system can also simultaneously encode a gene of TNF-related apoptosis-inducing ligand to suppress the primary tumor. Together, our study highlights the potential of local elimination of tumor pathogenic bacteria to prevent BC metastasis.

ObjectiveTo investigate the significance of different non-treatment thresholds or upper limits of normal （ULN） of alanine aminotransferase （ALT） in evaluating significant liver pathological injury in patients with chronic hepatitis B virus （HBV） infection， and to provide guidance for clinical diagnosis and treatment. MethodsThis study was conducted among 733 patients with chronic HBV infection who were hospitalized in Ningbo No. 2 Hospital from January 2015 to December 2023 and underwent liver biopsy and histopathological examination， and all patients had a persistent ALT level of ≤40 U/L and positive HBV DNA （>30 IU/mL）. According to the treatment threshold or ULN of ALT， the patients were divided into group 1 with 575 patients （≤35 U/L for male patients， ≤25 U/L for female patients）， group 2 with 430 patients （≤30 U/L for male patients， ≤19 U/L for female patients）， group 3 with 443 patients （≤27 U/L for male patients， ≤24 U/L for female patients）， group 4 with 446 patients （≤25 U/L）， group 5 with 158 patients （>35 U/L for male patients， >25 U/L for female patients）， and group 6 with 145 patients （>30 — ≤35 U/L for male patients， >19 — ≤25 U/L for female patients）. Groups 2， 5， and 6 were compared to analyze the severity of liver pathological injury in patients with different ALT levels and the constituent ratio of patients with significant liver pathological injury， and groups 1， 2， 3， and 4 were compared to investigate the value of different ULN or non-treatment thresholds of ALT in determining liver inflammation grade （G）， liver fibrosis stage （S）， and the treatment indication based on liver pathology. The independent-samples t test was used for comparison of normally distributed continuous data between two groups； a one-way analysis of variance was used for comparison between multiple groups， and the least significant difference t-test or the Tambane’s test was used for further comparison between two groups； the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups， and the Kruskal-Wallis H test was used for comparison between multiple groups and further comparison between two groups； the chi-square test or the Fisher’s exact test was used for comparison of categorical data between groups； a Ridit analysis was used for comparison of ranked data. A multivariate Logistic regression analysis （forward stepwise） was performed with whether liver pathology met the treatment indication （≥G2 and/or ≥S2） as the dependent variable and related factors with a significant impact on the dependent variable （P <0.05） as the independent variable. The receiver operating characteristic （ROC） curve was plotted， and the area under the ROC curve （AUC）， as well as sensitivity， specificity， positive predictive value， negative predictive value， positive likelihood ratio， and negative likelihood ratio， was used to assess the diagnostic value of different non-treatment thresholds of ALT. ResultsAmong the 733 patients， 259 （35.33%） had ≥G2 liver inflammation， 211 （28.79%） had ≥S2 liver fibrosis， and 306 （41.75%） had treatment indication （≥G2 and/or ≥S2）. There was a significant difference in liver inflammation grade （G0 — G4） between groups 2， 5， and 6 （χ²=22.869， P <0.001）， and there were also significant differences in the constituent ratios of patients with ≥G2 or ≥G3 liver inflammation between the three groups （χ²=21.742 and 14.921， P<0.001 and P=0.001）. There was a significant difference in liver fibrosis stage （S0 — S4） between groups 2， 5， and 6 （χ²=16.565， P<0.001）， and there were also significant differences in the constituent ratios of patients with ≥S2， ≥S3 or S4 liver fibrosis between the three groups （χ²=13.264， 13.050， and 6.260， P=0.001， 0.001， and 0.044）. There were significant differences between groups 2， 5， and 6 in the constituent ratios of patients with or without treatment indication based on liver pathology （χ²=20.728， P<0.001）. There were significant differences between groups 2， 5， and 6 in the constituent ratio of male patients （χ²=24.836， P<0.05）， age （F=5.710， P<0.05）， ALT （F=473.193， P<0.05）， aspartate aminotransferase （AST） （F=107.774， P<0.05）， ALT/AST ratio （F=40.167， P<0.05）， γ-glutamyl transpeptidase （GGT） （H=15.463， P<0.05）， aspartate aminotransferase-to-platelet ratio index （APRI） （H=63.024， P<0.05）， and LIF-5 （5 indicators for liver inflammation and fibrosis） （H=46.397， P<0.05）. In groups 1 — 4， compared with the patients without treatment indication， the patients with treatment indication had a significantly lower constituent ratio of patients with positive HBeAg， significantly lower levels of platelet count （PLT） and HBV DNA， and significantly higher age， ALT， AST， GGT， APRI， FIB-4， and LIF-5 （all P<0.05）. The Logistic regression analysis showed that age （odds ratio ［OR］=1.044， 95% confidence interval ［CI］： 1.025 — 1.063， P<0.001）， GGT （OR=1.022， 95%CI： 1.007 — 1.038， P=0.003）， and HBV DNA （OR=0.839， 95%CI： 0.765 — 0.919， P<0.001） were influencing factors for treatment indication based on liver pathology in group 1； HBeAg （OR=1.978， 95%CI： 1.269 — 3.082， P=0.003）， age （OR=1.048， 95%CI： 1.025 — 1.071， P<0.001）， GGT （OR=1.016， 95%CI： 1.001 — 1.031， P=0.041）， and PLT （OR=0.995， 95%CI： 0.991 — 1.000， P=0.049） were influencing factors in group 2； age （OR=1.040， 95%CI： 1.014 — 1.066， P=0.002）， ALT （OR=1.047， 95%CI： 1.005 — 1.092， P=0.029）， HBV DNA （OR=0.817， 95%CI： 0.736 — 0.907， P<0.001）， and LIF-5 （OR=7.382， 95%CI： 1.151 — 47.330， P=0.035） were influencing factors in group 3； age （OR=1.054， 95%CI： 1.031 — 1.077， P<0.001）， ALT （OR=1.061， 95%CI： 1.016 — 1.107， P=0.008）， and HBV DNA （OR=0.825， 95%CI： 0.743 — 0.917， P<0.001） were influencing factors in group 4. The diagnostic performance for identifying ≥G2 liver inflammation， ≥S2 liver fibrosis， and treatment indication in groups 1 — 4 had an AUC of >0.7； group 1 showed the lowest sensitivity （28.76%） and the highest specificity， positive predictive value， positive likelihood ratio， and negative likelihood ratio in judging treatment indication； group 2 had the highest sensitivity and negative predictive value and the lowest negative likelihood ratio； groups 3 and 4 had similar diagnostic indicators. ConclusionIn patients with chronic HBV infection and a persistently low ALT level， the severity of liver histopathological injury and the constituent ratio of significant liver histopathological injury decrease with the reduction in ALT level. A higher non-treatment threshold or ULN of ALT can help to identify the patients requiring treatment （with a higher specificity）， while a lower non-treatment threshold or ULN of ALT can help to identify the patients who do not require treatment （with a higher sensitivity）.

Objective To screen antidepressant-active compounds from Polygonati Rhizoma and explore their effects and possible mechanisms against depression induced by simulated weightlessness.Methods A systems pharmacology approach was used to screen potential antidepressant-active compounds and their targets from Polygonati Rhizoma.The hindlimb unloading(HLU)rat model was employed for the study.Twenty-four healthy male Sprague-Dawley rats were randomly divided into three groups:control group(administered 0.5%carboxymethylcellulose by gavage),HLU group(hindlimb unloading),and HLU+treatment group(hindlimb unloading+active compound gavage),with 8 rats in each group.After 28 days of hindlimb unloading,depressive-like behaviors in rats were evaluated using the forced swimming test and tail suspension test.Hippocampal morphology was examined with H&E staining,and GO and KEGG enrichment analyses were conducted on the targets of active compounds.Results A total of 38 active compounds were screened from Polygonati Rhizoma,among which apigenin had an oral bioavailability of 23.06%and a drug-likeness score of 0.21.Compound-target network analysis indicated that apigenin had the highest degree and betweenness centrality values,suggesting it might be the key active component with antidepressant potential in Polygonati Rhizoma.In the forced swimming and tail suspension tests,rats in the HLU group showed a significant increase in immobility time compared to the control group,indicating successful establishment of the depression model.However,compared to the HLU group,rats in the HLU plus apigenin group exhibited significantly reduced immobility time.The H&E staining results of hippocampal tissue showed a significant reduction in the number of hippocampal neurons,along with numerous shrunken neurons and small vacuoles in nerve fibers in the HLU group.In contrast,the treatment group exhibited an increased number of hippocampal neurons,with improved cellular morphology.Target enrichment analysis indicated that apigenin targets were mainly involved in the regulation of apoptosis and cancer-related signaling pathways.Conclusion Apigenin significantly improved depressive-like behaviors in rats subjected to hindlimb unloading,and it has a protective effect on hippocampal tissue.It may provide a new natural active compound for the treatment of depression caused by spaceflight-induced weightlessness.

Objective To investigate the analgesic effects of different doses of dexmedetomidine(Dex)in regulating the brain-derived neurotrophic factor(BDNF)/protein kinase B(AKT)/cyclica-denosine monophosphate response element binding protein(CREB)signaling pathway in elderly rats with spinal fractures.Methods Sixty rats were randomly divided into control group(skin incision only),model group(spinal fracture without any drug treatment),positive drug group(spinal fracture+0.2 μg/kg sufentanil),low-dose Dex group(spinal fracture+0.4 μg/kg Dex),medium-dose Dex group(spinal fracture+0.6 μg/kg Dex)and high-dose Dex group(spinal fracture+0.8 μg/kg Dex),with ten rats in each group.Mechanical pain threshold and thermal pain threshold were used to assess pain and sedation effects in elderly rats with spinal fractures.Modified neurological severity score(mNSS)was employed to evaluate the degree of neurological dysfunction.Enzyme-linked im-munosorbent assay(ELISA)was performed to detect serotonin(5-HT),substance P(SP),calci-tonin gene-related peptide(CGRP)neurotransmitter levels,tumor necrosis factor-α(TNF-α)and interleukin-1 β(IL-1 β)inflammatory factor levels.Colorimetric assays were used to measure malon-dialdehyde(MDA)oxidative stress expression levels;kits were applied to determine superoxide dis-mutase(SOD)and catalase(CAT)oxidative stress indicator levels.Western blotting(WB)analy-sis was conducted to examine BDNF,phosphorylated(p)-AKT and p-CREB protein expression lev-els in the spinal dorsal horn tissues of elderly rats with spinal fractures.Results Compared with the control group,the model group showed significantly increased mNSS scores,5-HT,SP,CGRP,TNF-α,IL-1 β and MDA expression levels,and significantly decreased mechanical pain thresholds,thermal pain thresholds,SOD,CAT,as well as BDNF,p-AKT and p-CREB protein expression lev-els(P＜0.05).Compared with the model group,the positive drug group and the medium-and high-dose Dex groups exhibited significantly lower mNSS scores,5-HT,SP,CGRP,TNF-α,IL-1βand MDA expression levels,and significantly higher mechanical pain thresholds,thermal pain thresholds,SOD,CAT,as well as BDNF,p-AKT and p-CREB protein expression levels(P＜0.05).Compared with the positive drug group,the low-dose Dex group had significantly higher mNSS scores,5-HT,SP,CGRP,TNF-α,IL-1 β and MDA expression levels,and significantly low-er mechanical pain thresholds,thermal pain thresholds,as well as BDNF,p-AKT andp-CREB pro-tein expression levels(P＜0.05);the high-dose Dex group showed significantly lower mNSS scores,5-HT,SP,CGRP,TNF-α,IL-1 β and MDA expression levels,and significantly higher me-chanical pain thresholds,thermal pain thresholds,as well as BDNF,p-AKT and p-CREB protein expression levels(P＜0.05).Compared with the low-dose Dex group,the medium-and high-dose Dex groups demonstrated significantly lower mNSS scores,5-HT,SP,CGRP,TNF-α,IL-1 β and MDA expression levels,and significantly higher mechanical pain thresholds,thermal pain thresh-olds,SOD,CAT,as well as BDNF,p-AKT and p-CREB protein expression levels(P＜0.05),in-dicating dose-dependent effect.Conclusion Dex may alleviate pain and reduce its impact on neu-rological function caused by spinal fractures in elderly rats through activation of the BDNF/AKT/CREB signaling pathway.

OBJECTIVE To explore the improvement effect and mechanism of petroleum ether extract of Saposhnikovia divaricata on rheumatoid arthritis （RA） rats by regulating neutrophil extracellular traps （NETs）. METHODS Establishment of rat RA model using bovine type Ⅱ collagen and Freund’s complete adjuvant. The model rats were randomly divided into model group and low-dose， middle-dose and high-dose groups （55， 110， 220 mg/kg） of petroleum ether extract of S. divaricata； the normal group without modeling was also established， with 10 rats in each group. Each group was given corresponding drugs or constant volume of 2% Tween-80 solution intragastrically， once a day， for consecutive 28 days. The toe swelling degree in rats was observed， and the arthritis index （AI） was scored. The serum levels of interleukin-1β （IL-1β）， IL-10， IL-17， tumor necrosis factor-α （TNF-α）， 25-hydroxyvitamin D3 [25（OH）D3]， myeloperoxidase （MPO）， neutrophil elastase （NE） and NETs in rats were detected. The histopathological changes in ankle joint were observed. The expression of citrullinated histone H3 （CitH3） in ankle joint as well as the expressions of cytochrome P450 24A1 （CYP24A1）， cytochrome P450 27B1 （CYP27B1），vitamin D receptor （VDR） and peptidyl arginine deiminase 4 疾病。E-mail：zhongyao626@126.com （PAD4） in synovium were all determined. RESULTS Compared with the model group， the toe swelling degree and AI score in the middle-dose and high-dose groups of petroleum ether extract of S. divaricata decreased significantly from day 14 to day 28 after administration （P＜0.05 or P＜0.01）. The serum levels of IL-1β， IL-17， TNF-α， MPO， NE and NETs decreased significantly， while the levels of IL-10 and 25（OH）D3 increased significantly （P＜0.05 or P＜0.01）. The widened ankle joint space and the improved structure were found； the expression of CitH3 in ankle joint， and the expressions of CYP24A1 and PAD4 in synovium were down-regulated significantly， while the expressions of CYP27B1 and VDR were up-regulated significantly in synovium （P＜0.05 or P＜0.01）. CONCLUSIONS The petroleum ether extract of S. divaricata may inhibit the production of NETs and improve the symptoms of RA by regulating the vitamin D system.

To develop a blue cap anticoagulant tube blood volume measuring card of to solve the problem of insufficient or excessive blood collection in clinical coagulation specimens.The device was composed of a measuring card,a transparent housing with a base and a tube holder.The measuring card was divided into qualified and unqualified areas,the housing was used to insert the card,the tube holder was used to place blood collection tubes.The device was used by clinical nurses to judge the adequacy of blood collection volume in blue cap anticoagulant tube.After the use of the device,the failure rate of clinical blue cap anticoagulation tube specimens submission was reduced from 6.71‰ to 2.73‰,shortened the time limit for specimen submission.At the same time,the device made the rejection judgment of department specimens more standardized and avoided the acceptance of unqualified specimens caused by subjective judgment errors.The device has simple structure,convenient operation and strong practicability,and has promotion value.

OBJECTIVE: To establish a machine learning model based on extreme gradient boosting (XGBoost) algorithm for early prediction of severe acute pancreatitis (SAP), and explore its predictive efficiency. METHODS: A retrospective cohort study was conducted. The patients with acute pancreatitis (AP) who admitted to the First Affiliated Hospital of Soochow University, the Second Affiliated Hospital of Soochow University and Changshu Hospital Affiliated to Soochow University from January 1, 2020 to December 31, 2021 were enrolled. Demography information, etiology, past history, and clinical indicators and imaging data within 48 hours of admission were collected according to the medical record system and image system, and the modified CT severity index (MCTSI), Ranson score, bedside index for severity in acute pancreatitis (BISAP) and acute pancreatitis risk score (SABP) were calculated. The data sets of the First Affiliated Hospital of Soochow University and Changshu Hospital Affiliated to Soochow University were randomly divided into training set and validation set according to 8 : 2. Based on XGBoost algorithm, the SAP prediction model was constructed on the basis of hyperparameter adjustment by 5-fold cross validation and loss function. The data set of the Second Affiliated Hospital of Soochow University was served as independent test set. The predictive efficacy of the XGBoost model was evaluated by drawing the receiver operator characteristic curve (ROC curve), and compared it with the traditional AP related severity score; variable importance ranking diagram and Shapley additive explanation (SHAP) diagram were drawn to visually explain the model. RESULTS: A total of 1 183 AP patients were enrolled finally, of which 129 (10.9%) developed SAP. Among the patients from the First Affiliated Hospital of Soochow University and Changshu Hospital Affiliated to Soochow University, there were 786 patients in the training set and 197 in the validation set; 200 patients from the Second Affiliated Hospital of Soochow University were used as the test set. Analysis of all three datasets showed that patients who advanced to SAP exhibited pathological manifestation such as abnormal respiratory function, coagulation function, liver and kidney function, and lipid metabolism. Based on the XGBoost algorithm, an SAP prediction model was constructed, and ROC curve analysis showed that the accuracy for prediction of SAP reached 0.830, the area under the ROC curve (AUC) was 0.927, which was significantly improved compared with the traditional scoring systems including MCTSI, Ranson, BISAP and SABP, the accuracy was 0.610, 0.690, 0.763, 0.625, and the AUC was 0.689, 0.631, 0.875, and 0.770, respectively. The feature importance analysis based on the XGBoost model showed that the top ten items ranked by the importance of model features were admission pleural effusion (0.119), albumin (Alb, 0.049), triglycerides (TG, 0.036), Ca²⁺ (0.034), prothrombin time (PT, 0.031), systemic inflammatory response syndrome (SIRS, 0.031), C-reactive protein (CRP, 0.031), platelet count (PLT, 0.030), lactate dehydrogenase (LDH, 0.029), and alkaline phosphatase (ALP, 0.028). The above indicators were of great significance for the XGBoost model to predict SAP. The SHAP contribution analysis based on the XGBoost model showed that the risk of SAP increased significantly when patients had pleural effusion and decreased Alb. CONCLUSIONS A SAP prediction scoring system was established based on the machine automatic learning XGBoost algorithm, which can predict the SAP risk of patients within 48 hours of admission with good accuracy.
Humans ; Pancreatitis ; Acute Disease ; Retrospective Studies ; Hospitalization ; Algorithms

Maintaining the stemness of the transplanted stem cell spheroids in an inflammatory microenvironment is challenging but important in regenerative medicine. Direct delivery of stem cells to repair periodontal defects may yield suboptimal effects due to the complexity of the periodontal inflammatory environment. Herein, stem cell spheroid is encapsulated by interfacial assembly of metal-phenolic network (MPN) nanofilm to form a stem cell microsphere capsule. Specifically, periodontal ligament stem cells (PDLSCs) spheroid was coated with Fe^III/tannic acid coordination network to obtain spheroid@[Fe^III-TA] microcapsules. The formed biodegradable MPN biointerface acted as a cytoprotective barrier and exhibited antioxidative, antibacterial and anti-inflammatory activities, effectively remodeling the inflammatory microenvironment and maintaining the stemness of PDLSCs. The stem cell microencapsulation proposed in this study can be applied to multiple stem cells with various functional metal ion/polyphenol coordination, providing a simple yet efficient delivery strategy for stem cell stemness maintenance in an inflammatory environment toward a better therapeutic outcome.
Anti-Bacterial Agents/pharmacology* ; Capsules/pharmacology* ; Cell Differentiation ; Cell Encapsulation ; Cells, Cultured ; Ferric Compounds/pharmacology* ; Osteogenesis/physiology* ; Periodontal Ligament ; Polyphenols/pharmacology* ; Stem Cells ; Tannins/pharmacology*

GuiLingJi(GLJ),a classic traditional Chinese medicine(TCM)formula,is composed of over 20 herbs,according to the Pharmacopeia of the People's Republic of China.Owing to its various activities,GLJ has been used in clinical settings for more than 400 years in China.However,the ambiguous chemical material basis limits the development of studies on the quality control and pharmacological mechanisms of GLJ.Therefore,comprehensive characterization of the multiple chemical components of GLJ is of great significance for the modernization of this formula.Given the great variety of herbs in GLJ,both UHPLC-MS and 1H NMR techniques were employed in this study.In addition,solvent extraction with different polarities was used to eliminate signal interference and the concentration of trace components.A variety of MS analytic methods were also used,including implementation of a self-built compound database,diagnostic ion filtering,mass defect filtering,and Compound Discoverer 3.0 analysis software.Based on the above strategies,a total of 150 compounds were identified,including 5 amino acids,13 phenolic acids and glycosides,11 coumarins,72 flavones,20 triterpenoid and triterpenoid saponins,23 fatty acids,and 6 other compounds.Moreover,13 compounds were identified by 1H NMR spectroscopy.The UHPLC-MS and 1H NMR results supported and complemented each other.This strategy provides a rapid approach to analyzing and identifying the chemical composition of Chinese herbal prescriptions.The current study provides basis for further research on the quality control and pharmacological mechanism of GLJ.

Objective:To establish and evaluate a new diagnostic model for significant liver tissue damage in patients with chronic hepatitis B virus (HBV) infection in the immune tolerance phase.Methods:The clinical data of 275 chronic HBV infection patients in the immune tolerance phase who underwent liver biopsy from January 2015 to November 2020 in the Hwa Mei Hospital, University of Chinese Academy of Sciences were included. According to the liver pathological changes, patients were divided into