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.

OBJECTIVE To form an expert consensus addressing clinical issues regarding the use of parenteral direct thrombin inhibitors （DTIs） in special populations. METHODS Led by the Sichuan Academy of Medical Sciences & Sichuan Provincial People’s Hospital（the Affiliated Hospital of UESTC）， a multidisciplinary working group was formed comprising experts from multiple fields， including clinical pharmacy， cardiac surgery， obstetrics， pediatrics and evidence-based medicine. Through literature review and the Delphi method， clinical questions regarding the efficacy and safety of parenteral DTIs used in special populations were identified. A structured design was adopted using the “Population-Intervention-Comparison-Outcome” （PICO） framework；systematic searches were conducted in CJFD， PubMed， Embase and other databases. Relevant evidence from randomized controlled trials，cohort studies and systematic reviews were included and synthesized. Evidence quality was assessed using the Grading of Recommendations Assessment，Development and Evaluation （GRADE） approach， and recommendations were formulated through three rounds of Delphi surveys and expert consensus meetings. RESULTS &CONCLUSIONS Seven clinical questions were ultimately selected （with a consensus rate exceeding 90%）， resulting in the formulation of seven recommendations on the use of parenteral DTIs in special populations， including children， pregnant women， patients with hepatic or renal impairment， patients with mesenteric venous thrombosis， and individuals with thrombophilia. These recommendations clarify the preferred agents， dosing ranges， monitoring parameters， and safety management strategies for parenteral DTIs in these special populations. This expert consensus， which is formulated based on the best available evidence， provides evidence-based guidance for standardized and individualized use of parenteral DTIs in special populations.

Immune-stimulating antibody drug conjugate (ISAC) can not only effectively solve the defects of stimulator of interferon genes (STING) agonists by coupling antibodies with STING agonists through the targeting of antibodies, but also play a synergistic role with antibodies to further improve the efficacy of STING agonists. This review first provides a concise overview of the current research landscape of ISACs and STING agonists, systematically elaborates on evolving trends in STING agonist development, and subsequently summarizes the mechanistic advances in STING ISAC research. Special emphasis is placed on representative STING ISAC candidates in preclinical/clinical development. Finally, the future directions of STING ISACs are critically discussed with perspectives and recommendations, aiming to provide theoretical insights and practical guidance for future investigations.

ObjectiveTo investigate the role and mechanism of Lck/Yes-related novel protein tyrosine kinase （Lyn） on lipopolysaccharide （LPS）-induced M1-type polarization of macrophage. MethodsThe LentiCRISPR-V2 plasmid was digested with the restriction endonuclease BSMBI-V2， and the digested DNA fragments were recovered. The digested plasmid was ligated with Lyn-sgRNA using T4 ligase to generate the Lenti-Lyn-gRNA lentivirus. THP-1 cells were infected with the Lenti-Lyn-gRNA lentivirus to obtain a stable cell line with Lyn knockout， and a monoclonal THP-1 cell line with complete Lyn knockout （Lyn^⁻/⁻） was established subsequently. Wild-type Lyn （Lyn^WT） and Lyn^⁻/⁻ THP-1 cells were induced with 100 ng/mL phorbol myristate acetate （PMA） for 48 h to differentiate into M0 macrophages， which were further polarized into M1 macrophages by stimulation with 100 ng/mL LPS for 24 h. Quantitative real-time polymerase chain reaction （qPCR） was performed to detect the expression of M0 macrophage markers， including integrin αM （CD11b）， macrophage antigen （CD68）， and monocyte differentiation antigen （CD14）. The expression of Lyn in M1 macrophages differentiated from wild-type THP-1 cells （Lyn^WT-M1） was measured by qPCR， and the ratio of phosphorylated Lyn to total Lyn （P-Lyn/Lyn） in Lyn^WT-M1 cells was determined by Western blot. In M1 macrophages differentiated from Lyn-knockout THP-1 cells （Lyn^⁻/⁻-M1）， qPCR was used to detect the mRNA expression of inducible nitric oxide synthase （iNOS）， interleukin-6 （IL-6）， and chemokine （C-X-C motif） ligand 10 （CXCL-10）. Western blot was conducted to assess the protein expression of iNOS， as well as the protein levels of molecules related to the Janus kinase 1 （JAK1）-signal transducer and activator of transcription 1 （STAT1）signaling pathway， including JAK1， phosphorylated JAK1 （P-JAK1）， STAT1， and phosphorylated STAT1 （P-STAT1）. Additionally， the expression of the M1 macrophage marker cluster of differentiation 80 （CD80） was analyzed by flow cytometry. ResultsThe Lyn^-/- monoclonal cell line was successfully constructed. The expression of CD11b was significantly elevated in Lyn^-/- M0 macrophages， and the differentiation of M1 macrophages was successful. Knockdown of Lyn inhibited mRNA expression of iNOS， IL⁃6， CXCL⁃10， protein expression of iNOS and CD80 expression in M1 macrophages （P<0.05）. Western blot assay showed that Lyn knockdown inhibited protein expression of JAK1 and P-STAT1 （P<0.01）. ConclusionAfter CRISPR/Cas9-mediated Lyn knockout， the expression levels of JAK1 and P-STAT1， the key molecules in the JAK/STAT signaling pathway of M1 macrophages， are significantly downregulated； concomitantly， the expression of M1 macrophage-specific secretory factors （iNOS， IL⁃6， CXCL⁃10 mRNA） and CD80 is also downregulated， which may be achieved via targeted regulation of the JAK1/P-STAT1-mediated JAK/STAT signaling pathway.

MicroRNA(miRNA)serves as a novel regulatory factor in viral and bacterial co-infections,exhibiting dual regulatory roles during pathogen infections.On the one hand,it exerts protective effects by modulating viral replication cycles and host anti-infection immune responses.On the other hand,it can be exploited by pathogens to promote infection progression,including influencing viral replication,antiviral immune responses,and regulating bacterial adhesion and proliferation.Mixed viral and bacterial infections lead to high mortality rates.By binding to specific target gene mRNA,miRNA degrades or inhibits its translation,thereby regulating target gene expres-sion,which may ultimately serve as potential diagnostic and therapeutic targets.This paper reviews recent ad-vances in research on the role of miRNA in viral and bacterial infections,particularly its mechanisms in co-infec-tions,aiming to provide theoretical reference for further research on the infection mechanisms.

Objective:To observe the applying effect of teamwork cooperation model of encephalopathy in the clinical teaching of Neurology.Methods:Sixty undergraduate students who practiced in our hospital were randomly divided into the control group and the experimental group,with 30 students in each group.The control group was arranged for internship according to the traditional department-based model,while the experimental group was arranged according to the teamwork cooperation model of encephalopathy discipline group.The theoretical examination scores and clinical skills scores of the two groups were compared,and the Mini-Clinical Evaluation Exercise(Mini-CEX)scale was used for dual evaluation by teachers and students from seven aspects:inquiry skills,physical examination skills,humanistic care,clinical judgment,communication skills,organizational effectiveness,and overall clinical competence.Result:There were statistically significant differences between the experimental group and the control group in theoretical examination scores,clinical skills scores,and dual evaluation by teachers and students(P＜0.05).Conclusion:Compared with the traditional teaching model,the teamwork cooperation model of encephalopathy discipline group has achieved better results in the clinical teaching of Neurology.

Purpose To explore the clinicopathologic features and differential diagnosis of nephrogenic adenomas(NAs)in urinary tract.Methods 27 cases of NA were collected.The clinicopathological features and differential di-agnosis were analyzed,and the relevant literatures were reviewed.Results A total of 27 cases of NA in urinary tract were included in this study,including 20 males and 7 females,ranging in age from 24 to 84 years.Among them,clini-cal manifestations included urinary tract irritation in 8 cases,hematuria in 7 cases,and no obvious symptoms or other comorbidities in 12 cases.Comorbidities:17 cases combined with urothelial tumors,4 cases with cystitis,2 cases with tuberculosis,2 cases with urethral diverticulum,and 2 cases with ureteral calculi or stenosis.Gross inspection:19 ca-ses occurred in the bladder,5 in the ureter,and 3 in the urethra.The average size of the lesions was about 4.3 mm.Local edema or protrusion of the mucosa with roughness were found in 18 cases,papillary or polypoid masses in 5 ca-ses,or follicular-like neoplasma in 4 cases.Histologic features:these lesions were mainly composed of papillary,tubu-lar and cystic structure,and eosinophilic colloid secretions were seen in the lumen.The lining epithelium was cuboidal or low columnar,some of which were 'hobnail' or signet ring-like cells,and stroma showed edema and chronic inflam-mation.Immunohistochemistry:epithelial cells expressed PAX8 and CK7,partially expressed P504S and GAT A3,but not p63 and PSA.p53 was wild type,and Ki67 proliferation index was low(1％-8％).Molecular testing:no ab-normalities were found in 14 urine FISH tests before surgery,and no fusion mutation was detected in 2 samples selected for second-generation sequencing.Treatment and follow-up:follow-up data were obtained for all cases,of which 24 un-derwent transurethral resection of bladder lesions,and the rest underwent nephroureterectomy or cystoprostatectomy.The mean follow-up was 26.6(1-59)months,and the lesion recurred in 2 cases.Conclusion NAs are relatively rare,and their final diagnosis depends on pathological examination.When the biopsy sample is small or the histomor-phology is atypical,it's often necessary to differentiate it from urothelial tumor,prostate adenocarcinoma and clear cell adenocarcinoma.Understanding the clinicopathologic and immunohistochemical features of NA is helpful to improve the diagnostic rate and avoid missed-diagnosis or misdiagnosis.

Objective To explore the mechanism of honey-processed Hedysari Radix in the regulation of intestinal immunity in rats with spleen qi deficiency,which was based on G protein-coupled receptor 41(GPR41)/GPR43-mediated mitogen-activated protein kinase(MAPK)signaling pathway.Methods The three-factor composite modeling method of eating disorder,diarrhea and fatigue was used to establish a model of spleen qi deficiency,and the rats were randomly divided into model,honey-processed Hedysari Radix,probiotics and blank groups with 15 rats per group.The honey-processed Hedysari Radix group was given by gavage 12.6 g·kg-1 aqueous extract of honey-processed Hedysari Radix.The probiotics group was given 0.625 g·kg-1 bifidobacterium triple viable solution by gavage.The blank and model groups were given the same dose of distilled water by gavage.Four groups were treated for 15 d with once a day.The expression levels of GPR41,GPR43,P38 MAPK,c-Jun N-terminal kinase(JNK)and extracellular regulatory protein kinase 1/2(ERK1/2)in colon tissues were detected by Western blotting.Results The relative expression levels of GPR41 in the blank,model,honey-processed Hedysari Radix and probiotics groups were 0.95±0.07,0.45±0.03,0.84±0.19 and 0.86±0.20;the relative expression levels of GPR43 were 1.17±0.11,0.41±0.06,0.66±0.03 and 0.57±0.01;the phosphorylated ERK1/2/ERK1/2 ratios were 0.16±0.01,0.43±0.01,0.39±0.01 and 0.36±0.02;the phosphorylated JNK/JNK ratios were 0.58±0.05,1.47±0.10,0.90±0.11 and 0.90±0.11;the phosphorylated P38 MAPK/P38 MAPK ratios were 1.77±0.33,3.19±0.03,2.01±0.17 and 2.23±0.59,respectively.Compared with the model group,the differences of above indexes were statistically significant in the honey-processed Hedysari Radix and probiotics groups(P＜0.05,P＜0.01).Conclusion The mechanism of honey-processed Hedysari Radix regulating intestinal immunity in rats with spleen qi deficiency is related to the regulation of GPR41/GPR43 mediated MAPK signaling pathway.

Objective To explore the lung protective effect of pressure controlled ventilation-volume guaranteed(PCV-VG)combined with driving pressure(DP)guided lung protective ventilation strategy in patients undergoing thoracoscopic and laparoscopic radical esophagectomy.Methods 70 patients scheduled for elective thoracoscopic and laparoscopic radical esophagectomy were allocated into two groups using a random number table method:Conventional lung protective ventilation strategy group(group C)and DP guided lung protective ventilation strategy under PCV-VG mode group(group P),35 case in each group.Peak airway pressure(Ppeak),plateau pressure(Pplat),dynamic compliance(Cdyn)and DP were compared between the two groups at 5 minutes after intubation(T1),30 min after pneumoperitoneum established(T2),just prior to one lung ventilation(OLV)(T3),30 min after OLV(T4),60 min after OLV(T5)and 15 min from recovery of two lung ventilation(TLV)(T6).The blood pressure(BP),heart rate(HR),arterial partial pressure of oxygen(PaO2),partial pressure of carbon dioxide in arterial blood(PaCO2)and pH were recorded before anesthesia(T0),T2,T3,T4,T5 and T6 time points.The occurrence of postoperative pulmonary complications(PPCs)also recorded.Results Compared with group C,Ppeak in group P at T1,T2,T4,T5 and T6 time points was significantly decreased,and Cdyn was obviously increased,the differences were statistically significant(P＜0.05).At the T1,T4,T5 and T6 time points,the DP was lower in group P compared to group C,and Pplat at T6 time point was lower than that in group C,the differences were statistically significant(P＜0.05).At the time points of T4 and T5,the PaO2 in group P was higher than that in Group C,and the PaCO2 at T6 time point was also higher than that in group C,the differences were statistically significant(P＜0.05).The comparison of PaCO2 at T0,T2,T3,T4 and T5 time points of the two groups,the difference was not statistically significant(P＞0.05).Comparison of pH between the two groups,the difference was not statistically significant at all time points(P＞0.05).The systolic blood pressure(SBP)of group P was higher than that of group C at the T4 time point,and the diastolic blood pressure(DBP)was lower than that of group C at T6 time point,and the differences were statistically significant(P＜0.05);There were no significant differences in SBP and DBP at T0,T2,T3 and T5 time points,and HR at each time point between the two groups(P＞0.05).There was no statistically significant difference in the occurrence of PPCs within 7 d after operation between the two groups(P＞0.05).Conclusion DP guided lung protective ventilation strategy under PCV-VG mode can improve intraoperative respiratory mechanics,and increase oxygenation during OLV in patients undergoing thoracoscopic and laparoscopic radical esophagectomy,but it does not significantly affect the incidence of PPCs within 7 d after operation.It is worthy clinical significant.