ObjectiveThis paper aims to find the identified and validated clinical biomarker data building upon a clinical study of early-phase phase Ⅱ and investigate the correlation analysis of Huanglian Jiedu Wan on syndrome improvement and clinical biomarkers in the treatment of "excess heat-toxicity" based on a machine learning model. Additionally， the effective prediction of clinical biomarker values for the main symptoms of the "excess heat-toxicity" syndrome was assessed. MethodsA total of 229 patients meeting the inclusion criteria for "excess heat-toxicity" syndrome were randomly divided into the Huanglian Jiedu Wan group and the placebo group. Syndrome score transition matrices were constructed for the Huanglian Jiedu Wan group and the placebo group based on three main symptoms of "excess heat-toxicity" syndrome， such as oral ulcers， sore throat， and gum swelling and pain. Data from the patients with these three syndromes were also integrated for an overall analysis. The corresponding syndrome score transition matrices were further constructed to visualize symptom change trends of the patients in the two groups via heatmaps. Based on the identified and validated clinical biomarkers related to inflammation， oxidative stress， and energy metabolism in the early phase， Spearman correlation analysis was employed to analyze and evaluate the associations between clinical biomarkers and syndrome improvement. Key clinical biomarkers reflecting the effect of Huanglian Jiedu Wan were screened through the comparison of differences between groups. An extreme gradient boosting （XGBoost） algorithm was used to develop a prediction model for main symptom classification， with classification performance evaluated through 10-fold cross-validation. Feature importance analysis was applied to identify variables with the greatest contribution to the prediction result. ResultsThe syndrome transition matrix results indicated that the Huanglian Jiedu Wan group showed a superior effect to the placebo group in improving oral ulcers， sore throat， and overall symptoms， with significant effects observed especially in sore throat and overall symptom analyses （P<0.01）. Spearman correlation analysis revealed that several clinical biomarkers positively correlated with "excess heat-toxicity" syndrome and its main symptom improvement， were also called "heat-related biomarkers"， including succinic acid， α-ketoglutaric acid， glycine， lactic acid， adenosine monophosphate （AMP）， tumor necrosis factor-α （TNF-α）， interferon-γ （IFN-γ）， interleukin-1β （IL-1β）， interleukin-4 （IL-4）， interleukin-6 （IL-6）， interleukin-8 （IL-8）， interleukin-10 （IL-10）， and so on. Conversely， clinical biomarkers negatively correlated with symptom severity， were also called "heat-clearing related biomarkers" after administration of Huanglian Jiedu Wan， including malic acid， fumaric acid， cis-aconitic acid， adrenocorticotropic hormone （ACTH）， IL-1β， IL-4， IL-8， succinic acid， and citric acid. The XGBoost classification model using all 52 biomarkers as variables achieved an average test accuracy of 0.754 and an average F1 score of 0.777. Feature importance analysis identified the scores of glutamic acid in saliva and IL-6 were the highest in all the variables， with importance scores of 0.081 and 0.080， respectively. After screening out 14 key variables and optimizing the parameters， model performance improved to an average accuracy of 0.758 and an F1 score of 0.798. Feature importance analysis further determined that the glutamic acid in saliva and IL-6 showed obvious changes after screening the variables， confirming the good syndrome prediction ability of the model constructed by these key clinical biomarkers. ConclusionThis study systematically elucidates the correlation between syndrome improvement and clinical biomarkers of Huanglian Jiedu Wan in the treatment of "excess heat-toxicity" syndrome. An XGBoost classification model based on key clinical biomarkers is successfully established， achieving effective prediction of the symptoms related to the "excess heat-toxicity" syndrome such as oral ulcers and sore throat and providing a new insight for objective identification of traditional Chinese medicine syndromes.

ObjectiveThis paper aims to find the identified and validated clinical biomarker data building upon a clinical study of early-phase phase Ⅱ and investigate the correlation analysis of Huanglian Jiedu Wan on syndrome improvement and clinical biomarkers in the treatment of "excess heat-toxicity" based on a machine learning model. Additionally， the effective prediction of clinical biomarker values for the main symptoms of the "excess heat-toxicity" syndrome was assessed. MethodsA total of 229 patients meeting the inclusion criteria for "excess heat-toxicity" syndrome were randomly divided into the Huanglian Jiedu Wan group and the placebo group. Syndrome score transition matrices were constructed for the Huanglian Jiedu Wan group and the placebo group based on three main symptoms of "excess heat-toxicity" syndrome， such as oral ulcers， sore throat， and gum swelling and pain. Data from the patients with these three syndromes were also integrated for an overall analysis. The corresponding syndrome score transition matrices were further constructed to visualize symptom change trends of the patients in the two groups via heatmaps. Based on the identified and validated clinical biomarkers related to inflammation， oxidative stress， and energy metabolism in the early phase， Spearman correlation analysis was employed to analyze and evaluate the associations between clinical biomarkers and syndrome improvement. Key clinical biomarkers reflecting the effect of Huanglian Jiedu Wan were screened through the comparison of differences between groups. An extreme gradient boosting （XGBoost） algorithm was used to develop a prediction model for main symptom classification， with classification performance evaluated through 10-fold cross-validation. Feature importance analysis was applied to identify variables with the greatest contribution to the prediction result. ResultsThe syndrome transition matrix results indicated that the Huanglian Jiedu Wan group showed a superior effect to the placebo group in improving oral ulcers， sore throat， and overall symptoms， with significant effects observed especially in sore throat and overall symptom analyses （P<0.01）. Spearman correlation analysis revealed that several clinical biomarkers positively correlated with "excess heat-toxicity" syndrome and its main symptom improvement， were also called "heat-related biomarkers"， including succinic acid， α-ketoglutaric acid， glycine， lactic acid， adenosine monophosphate （AMP）， tumor necrosis factor-α （TNF-α）， interferon-γ （IFN-γ）， interleukin-1β （IL-1β）， interleukin-4 （IL-4）， interleukin-6 （IL-6）， interleukin-8 （IL-8）， interleukin-10 （IL-10）， and so on. Conversely， clinical biomarkers negatively correlated with symptom severity， were also called "heat-clearing related biomarkers" after administration of Huanglian Jiedu Wan， including malic acid， fumaric acid， cis-aconitic acid， adrenocorticotropic hormone （ACTH）， IL-1β， IL-4， IL-8， succinic acid， and citric acid. The XGBoost classification model using all 52 biomarkers as variables achieved an average test accuracy of 0.754 and an average F1 score of 0.777. Feature importance analysis identified the scores of glutamic acid in saliva and IL-6 were the highest in all the variables， with importance scores of 0.081 and 0.080， respectively. After screening out 14 key variables and optimizing the parameters， model performance improved to an average accuracy of 0.758 and an F1 score of 0.798. Feature importance analysis further determined that the glutamic acid in saliva and IL-6 showed obvious changes after screening the variables， confirming the good syndrome prediction ability of the model constructed by these key clinical biomarkers. ConclusionThis study systematically elucidates the correlation between syndrome improvement and clinical biomarkers of Huanglian Jiedu Wan in the treatment of "excess heat-toxicity" syndrome. An XGBoost classification model based on key clinical biomarkers is successfully established， achieving effective prediction of the symptoms related to the "excess heat-toxicity" syndrome such as oral ulcers and sore throat and providing a new insight for objective identification of traditional Chinese medicine syndromes.

BACKGROUND: Arsenic trioxide (ATO) is indicated as a broad-spectrum medicine for a variety of diseases, including cancer and cardiac disease. While the role of ATO in hepatic ischemia/reperfusion injury (HIRI) has not been reported. Thus, the purpose of this study was to identify the effects of ATO on HIRI. METHODS: In the present study, we established a 70% hepatic warm I/R injury and partial hepatectomy (30% resection) animal models in vivo and hepatocytes anoxia/reoxygenation (A/R) models in vitro with ATO pretreatment and further assessed liver function by histopathologic changes, enzyme-linked immunosorbent assay, cell counting kit-8, and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay. Small interfering RNA (siRNA) for extracellular signal-regulated kinase (ERK) 1/2 was transfected to evaluate the role of ERK1/2 pathway during HIRI, followed by ATO pretreatment. The dynamic process of autophagic flux and numbers of autophagosomes were detected by green fluorescent protein-monomeric red fluorescent protein-LC3 (GFP-mRFP-LC3) staining and transmission electron microscopy. RESULTS: A low dose of ATO (0.75 μmol/L in vitro and 1 mg/kg in vivo ) significantly reduced tissue necrosis, inflammatory infiltration, and hepatocyte apoptosis during the process of hepatic I/R. Meanwhile, ATO obviously promoted the ability of cell proliferation and liver regeneration. Mechanistically, in vitro  studies have shown that nontoxic concentrations of ATO can activate both ERK and phosphoinositide 3-kinase-serine/threonine kinase (PI3K-AKT) pathways and further induce autophagy. The hepatoprotective mechanism of ATO, at least in part, relies on the effects of ATO on the activation of autophagy, which is ERK-dependent. CONCLUSION Low, non-toxic doses of ATO can activate ERK/PI3K-AKT pathways and induce ERK-dependent autophagy in hepatocytes, protecting liver against I/R injury and accelerating hepatocyte regeneration after partial hepatectomy.
Animals ; Arsenic Trioxide ; Autophagy/physiology* ; Reperfusion Injury/prevention & control* ; Mice ; Male ; Proto-Oncogene Proteins c-akt/physiology* ; Arsenicals/therapeutic use* ; Oxides/therapeutic use* ; Liver/metabolism* ; Extracellular Signal-Regulated MAP Kinases/metabolism* ; Mice, Inbred C57BL

Objective:To explore the influences of FibroScan detection on the pregnancy outcomes of pregnant rats transfected with hepatitis B virus and the growth of their offspring rats.Methods:Sixty SPF-grade SD rats with a male-to-female ratio of 5∶1 were selected，randomly divided the female rats into the groups of mid-pregnancy with hepatitis B，late-pregnancy with hepatitis B，normal mid-pregnancy，normal late-pregnancy，and control group，and the rats in the mid-pregnancy and late-pregnancy groups were subjected to FibroScan testing on the 10th and 15th days of gestation respectively. The number of offspring rats in each group was randomly reduced to 10 on the 3rd day after delivery，and the offspring rats were divided into the groups of mid-pregnancy with hepatitis B，late-pregnancy with hepatitis B，normal mid-pregnancy，normal late-pregnancy，and control group. The pregnancy outcome levels of pregnant rats were observed by adopting a stratified analysis strategy，including differences in weight changes，number of deliveries，fetal rat outcomes，lactation conditions，litter size，and litter weight. The growth levels of offspring rats，including differences in weight，body length，tail length，and the time of fur growth，tooth eruption，eye opening，and ear standing on the 21st day were observed，and the survival rate of the offspring rats was compared.Results:With regard to the pregnancy outcome levels of the pregnant rats，there were no statistically significant differences in weight changes during pregnancy among the five groups of pregnant rats［ F（4，45）=2.627，Adjusted P=0.222］. There were also no statistically significant differences in the number of deliveries，fetal rat outcomes，and lactation conditions（Adjusted P=1.000）. The number of deliveries［ F（4，21.095）=2.280，Adjusted P=0.222］and litter weight［ F（4，20.128）=2.159，Adjusted P=0.222］showed no statistically significant differences. After correction using the BH method，none of the indicators showed significant differences（ P>0.05）. In terms of the growth levels of the offspring rats，among the five groups of offspring rats，there were no statistically significant differences in body weight on the 21st day［H（4）=11.623，Adjusted P=0.135］，body length on the 21st day［H（4）=10.962，Adjusted P=0.135］，and tail length on the 21st day［H（4）=9.126，Adjusted P=0.058］. Besides，the differences in the time of fur growth，tooth eruption，eye opening，and ear standing［H（4）=0.000，Adjust P=1.000］showed no statistical significance. The survival rate on the 7th day，14th day，and 21st day was 100%，with no significant differences（Adjusted P=1.000）. After correction using the BH method，none of the indicators showed significant differences（ P>0.05）. Conclusion:FibroScan detection had no significant influences on the pregnancy outcomes of pregnant rats with hepatitis B or on the growth of their offspring rats in multiple stages，dimensions，and indicators，indicating that FibroScan detection is safe for pregnant rats. This research provides an animal experimental basis for the safe use of FibroScan in pregnant women with chronic HBV infection.

Objective:To establish a new method and platform for screening, identifying, and exploring a new strategy for anti-hepatitis B immunotherapy based on hepatitis B virus (HBV)-specific TCR.Methods:Peripheral blood mononuclear cells were isolated from patients with acute hepatitis B. CD3 +CD8 +CD137 +T single cells were sorted out after stimulation with the HBsAg peptide library. The α and β chains in TCRs of single cells were amplified by PCR. TCR double-chain pairing and lentiviral packaging were performed through high-throughput sequencing. Re-infected Jurkat-76-NFAT-GFP cells and the cell lines stably expressing TCR were screened. HBsAg peptide library and immortalized B lymphocytes co-cultured with J76N-TCR were used to screen HBsAg-specific TCRs. K562 cell lines stably expressing HLA-A*24:02 were established to determine epitope peptide by screening A*24:02-restricted TCR. The screened TCRs were replaced with mouse C regions and packaged with lentiviruses. Functional validation was performed on healthy human CD4 +T and CD8 +T lymphocytes following infection. Results:Stable TCR-expressing cell lines were successfully prepared based on single-cell TCRαβ double-chain amplification and pairing technology. Twenty-one TCRs were screened using immortalized B lymphocytes, resulting in nine possible HLA-A*24:02-restricted HBsAg-specific TCRs. Further screening with K562-A2402 resulted in six A*24:02-restricted HBsAg-specific TCRs with identically recognized epitope peptide. The functional determination of the two TCR clones revealed their specific recognition function for target cells expressing HBsAg.Conclusion:HLA-A*24:02-restricted HBsAg-specific TCR with recognition function for target cells expressing HBsAg was successfully obtained based on the new experimental technology system, laying an important foundation for further exploration of antiviral immunotherapy based on HBV-specific TCR.

Objective:To explore the abnormal patterns of brain functional network connectivity in depression adolescents and their diagnostic value in adolescent depression.Methods:A total of 94 depression adolescents (adolescent depression group) admitted to Outpatient Department of Psychiatric Imaging, West China Hospital, Sichuan University from January 2020 to December 2022 were selected. In addition, 78 age- and gender-matched healthy adolescents were recruited from local community advertisements at the same time-period as healthy control group. Resting-state functional magnetic resonance imaging was performed; after image preprocessing, group-level spatial independent component analysis was performed to identify the intrinsic network connectivity, and differences in network connectivity between the two groups were compared. Functional connectivity edges were employed as classification features, and feature ranking and screening were then performed. A support vector machine (SVM) with linear kernel function was used to construct a classification model, and receiver operating characteristic (ROC) curve was used to analyze the diagnostic value of this classification model in adolescent depression.Results:No significant difference was noted in age, gender, years of education, and body mass index between the two groups ( P>0.05). Compared with the healthy control group, the adolescent depression group had significantly decreased functional connectivity intensity within and between the networks of sensorimotor network (SMN), visual network (VN), auditory network (AN), default mode network (DMN), and cognitive control network (CCN), and significantly increased functional connectivity intensity within CCN ( P<0.05). When using the 75 top-ranked functional connectivity features, this classification model had the best performance (accuracy rate: 70.35%, sensitivity: 70.21%, specificity: 71.80%, P<0.001). ROC curve showed that area under the curve of this classification model in diagnosing adolescent depression was 0.724 (95% CI: 0.648-0.800, P<0.001). A total of 51 consistent functional connectivities were identified and they were mainly located within or between the networks of SMN, VN, AN, DMN, and CCN. Conclusion:The abnormal resting-state brain functional connectivity in depression adolescents can provide imaging basis for their clinical diagnosis.

AIM:This study aims to investigate the role of ferroptosis in the myocardium of mice with lipopoly-saccharide(LPS)-induced sepsis and in the injury of H9c2 rat cardiomyocytes,and to explore the regulatory function of microRNA-351-5p(miR-351-5p)in this context.METHODS:An in vivo model of sepsis-induced cardiomyopathy was established in mice through intraperitoneal injection of LPS.Twenty-four mice were randomly divided into negative control(NC)group,LPS group,and LPS+ferroptosis inhibitor ferrostatin-1(Fer-1)group.Hematoxylin-eosin(HE)staining was conducted to assess cardiac injury,and plasma levels of creatine kinase(CK)and lactate dehydrogenase(LDH)were also measured.Additionally,the levels of Fe2+and malondialdehyde(MDA)in plasma were quantified,and the mRNA levels of acyl-CoA synthetase long chain family member 4(ACSL4)and prostaglandin-endperoxide synthase 2(PTGS2)were de-tected by RT-qPCR.In vitro,H9c2 cardiomyocytes were stimulated with LPS to create cellular models,followed by treat-ment with Fer-1,inhibitor NC,or miR-351-5p inhibitor.Cell viability was evaluated using CCK8 assay,intracellular re-active oxygen species(ROS)were measured by flow cytometry,intracellular Fe2+levels were assessed using a fluorescence probe,and the protein expression of glutathione peroxidase 4(GPX4)and ACSL4 was analyzed by Western blot.The MDA and reduced glutathione(GSH)levels were measured using commercial kits.MicroRNA(miRNA)sequencing was performed on the LPS-stimulated H9c2 cardiomyocyte models,with differential miRNAs identified and subsequently vali-dated using RT-qPCR.RESULTS:The mice in LPS group exhibited significant myocardial tissue dysregulation com-pared with NC group,with enlarged space,increased plasma CK and LDH levels(P<0.05),elevated Fe2+and MDA levels in myocardial tissues(P<0.05),and increased mRNA levels of ACSL4 and PTGS2(P<0.05).In contrast,the mice in LPS+Fer-1 group demonstrated improved myocardial tissue structure,reduced space,decreased plasma CK and LDH levels(P<0.05),and lower Fe2+and MDA levels in myocardial tissues(P<0.05),along with decreased mRNA level of PTGS2(P<0.05).In H9c2 cardiomyocytes,cell viability,intracellular GSH level,and GPX4 protein level were significantly reduced in LPS group compared with NC group(P<0.05),while ROS,MDA,Fe2+,and ACSL4 protein levels were elevated(P<0.05).The cells in LPS+Fer-1 group showed increased viability,intracellular GSH level,and GPX4 protein level compared with LPS group(P<0.05),alongside reduced ROS,MDA,Fe2+,and ACSL4 levels(P<0.05).miRNA sequencing revealed a significant decrease in several miRNAs,with miR-351-5p showing the most pro-nounced reduction.In LPS+miR-351 inhibitor group,H9c2 cell viability significantly declined(P<0.05),and the levels of GSH and GPX4 were notably lowered(P<0.05),while ROS,MDA,Fe2+and ACSL4 protein levels were significantly elevated(P<0.05).However,in LPS+miR-351 inhibitor+Fer-1 group,the cell viability increased(P<0.05),and the GSH level rose significantly(P<0.05),with corresponding decreases in intracellular ROS,Fe2+and ACSL4 protein levels(P<0.05).CONCLUSION:Inhibition of ferroptosis attenuated sepsis-induced myocardial injury,and inhibition of miR-351-5p promotes sepsis-induced ferroptosis of H9c2 cardiomyocytes.

Objective To investigate the association between gout-related gene polymorphisms and clinical phenotypic heterogeneity among gout patients in the Foshan region,thereby providing a scientific basis for stratified clinical management.Methods A total of 125 gout patients diagnosed at the Foshan Hospital of Traditional Chinese Medicine between June 2022 and May 2025 were enrolled in this study.The collected data included demo-graphic characteristics,frequency of gout attacks,presence of tophi,levels of uric acid,creatinine,C-reactive protein(CRP),erythrocyte sedimentation rate(ESR),gout-related genes(ABCG2,SLC2A9,SLC22A12,MTHFR),and joint ultrasound findings.Group comparisons and rank correlation analyses were conducted to explore potential associations between gene polymorphisms and clinical heterogeneity.Results The male-to-female ratio was 11∶1;the mean age was(35.28±2.67)years;the mean disease duration was(6.03±0.68)years;and the mean frequency of acute attacks in the past 12 months was 4(2.0,7.25).Genotype distributions were as follows:ABCG2:wild-type(C/C),23.8%;heterozygous(C/A),53.2%;homozygous(A/A),23%.SLC2A9:wild-type(A/A),24.6%;heterozygous(A/G),50%;homozygous(G/G),25.4%.SLC22A12:wild-type(A/A),4.8%;heterozygous(A/C),31.7%;homozygous(C/C),63.5%.MTHFR:wild-type(C/C),68.3%;heterozygous(C/T),28.6%;homozygous(T/T),3.2%.Rank correlation analysis revealed that SLC2A9 polymorphisms were significantly correlated with tophi formation(ρ=0.193,P=0.031)and crystal deposition on ultrasound(ρ=0.202,P=0.025).SLC22A12 polymorphisms were associated with hypertension(ρ=0.269,P=0.003)and diabetes(ρ=0.200,P=0.026).MTHFR polymorphisms showed a correlation with diabetes(ρ=0.224,P=0.012).Conclusions Polymorphisms in SLC2A9,SLC22A12,and MTHFR are significantly linked to clinical phenotypic heterogeneity among gout patients.Genetic testing could facilitate the early identification of individuals at high risk for complications and support the development of stratified and individualized treatment approaches.

Objective To explore the identification method,pathogenesis,clinical characteristics and individualized pharmacotherapy of asymptomatic hyperuricemia after renal transplantation.Methods The pharmacist was on duty at the organ transplant outpatient clinic.During this time,they analyzed and sorted out the medications,identified and differentiated a case of asymptomatic hyperuricemia related to spironolactone in a patient who had undergone a renal transplant,and provided comprehensive care throughout the entire process.Results The asymptomatic hyperuricemia in this patient might be associated with spironolactone,and the adverse reactions of the patient were alleviated by pharmacists through optimizing clinical treatment.Up to now,no hyperuricemia occurred.Conclusions Pharmacists are required to collaborate closely with clinicians to establish medication profiles for patients under long-term follow-up and to closely monitor and evaluate drug-related adverse reactions.Additionally,they should assess the renal function and immune status of transplant recipients promptly and formulate individualized treatment plans in order to enhance the long-term survival of both the transplanted kidneys and the recipients.

Objective:To integrate the symptom management experiences of patients with acute decompensated heart failure (ADHF), so as to provide a basis for developing symptom management measures.Methods:Qualitative or mixed studies on the symptom management experience of ADHF patients included from establishment of the database to September 2024, were electronically retrieved in PubMed, CINAHL, Cochrane Library, Web of Science, Embase, EBSCO, China National Knowledge Infrastructure, WanFang Data, VIP, China Biomedical Database and other Chinese and English databases and gray literature databases. The quality of the literature was evaluated using the Critical Appraisal Skills Programme developed by the Center for Evidence-Based Medicine at the University of Oxford. The results were synthesized through the aggregative integration method.Results:A total of 14 papers were included. Thirty-four findings were distilled into eight categories and three integrative findings, namely, the multiple challenges posed by symptoms (complex and multiple symptomatic somatic experiences, symptom-induced mood changes, and reduced family and social engagement), the unmet needs of patients (insufficient healthcare resources, insufficient supply of discharge services provided by healthcare organizations, and lack of knowledge), and the co-existence of positive and negative coping styles (negative coping styles in symptomatic distress, positive debugging and diversified coping in symptomatic distress) .Conclusions:ADHF symptoms severely affect patients' physical, psychological, and social function. Healthcare professionals should focus on the unmet needs of patients with ADHF and explore patient-engaged models of active symptom management.