ObjectiveTo evaluate the detection specificity for clinical samples and the detection capability for standard substances of five commercially available multiplex polymerase chain reaction (PCR) nucleic acid detection kits (hereinafter referred to as the kits) for respiratory pathogens, and to provide a reference for selecting appropriate detection kits for multi-pathogen nucleic acid testing of respiratory infections. MethodsA total of 60 respiratory pathogen-positive clinical samples with known redults were selected and tested using the five kits (labeled as A, B, C, D, and E). The detection rates and Kappa coefficients were calculated to evaluate the consistency between the results from these kits and those from single-pathogen PCR kits. According to the limit of detection (LOD) provided by the kits, standard substances of respiratory pathogens (including 12 types such as influenza virus, Mycoplasma pneumoniae, and Bordetella pertussis) were diluted to four concentrations (250, 500, 1 000, and 2 000 copies·mL⁻¹). All five kits were used for detection to evaluate their respective detection capabilities. ResultsCompared with the results from single-pathogen PCR kits, the five tested kits demonstrated good consistency (all Kappa >0.80). Among them, Kit A had the highest detection rate (100.00%), followed by Kits C and E (98.33%), and then Kits B and D (95.00%). All five kits showed a relatively low false negative rate (FNR) for samples with a cycle threshold (Ct) value ≤35 (≤2.38%). However, for samples with Ct values>35, the FNR increased accordingly(average FNR=6.67%, P=0.029). Kit C exhibited the highest detection sensitivity for the tested standard substances (average LOD: 458.33 copies·mL⁻¹), followed by Kit D, then Kits A/E, and finally Kit B. ConclusionThe five multiplex PCR kits showed good consistency with single-pathogen detection results, but each had its own performance emphasis. Kit A, with the highest detection rate and high throughput, is suitable for targeted viral screening. Kit B, covering the broadest pathogen spectrum (including fungi/bacteria), is suitable for comprehensive respiratory pathogen screening. Kits C, D and E, are applicable for rapid detection. It is important to note that the detection efficacy of all kits decreases for low viral load samples with Ct values >35. In practical application, selection should be based on specific screening objectives, throughput requirements, and sample types.

ObjectiveThe most prevalent mRNA modification, N6-methyladenosine (m⁶A) plays an important role in various RNA metabolism, including gene expression and translation. By recruiting different “reader” proteins and their cofactors, m⁶A modification can affect messenger RNA (mRNA) degradation, splicing, nuclear export and translation. However, the selective mechanism by which m⁶A sites regulate mRNA translation through m⁶A reader YTHDF1 binding remains poorly understood, due to a lack of computational methods for identifying context-specific m⁶A sites that regulate translation. To address this, we developed a novel computational framework named m⁶ATEpre, the first tool designed to predict cell-specific m⁶A sites that regulate translation efficiency. Methodsm⁶ATEpre integrates multi-omics data, introduces a novel feature representation strategy for m⁶A site sequences, and employs an autoencoder to effectively capture embedded feature representations. Specifically, m⁶ATEpre first integrated MeRIP-seq data and PAR-CLIP data through overlapping m⁶A sites with YTHDF1 binding sites and identified YTHDF1-mediated m⁶A sites. Then, m⁶ATEpre detected the translation gene by analyzing the Ribo-seq data under YTHDF1 knockdown vs control condition. Genes whose translation is mediated by YTHDF1 in an m⁶A-dependent manner were identified by a significant decrease in translation efficiency upon YTHDF1 knockdown. Next, we proposed a binary vector indicating the presence or absence of YTHDF1 binding motifs to characterize each m⁶A site sequence. This represents a novel feature representation strategy for m⁶A sites. m⁶ATEpre utilized the autoencoder to extract the potentially important feature representations and constructed a multilayer perceptron neural networks model to predict potential m⁶A sites that regulating translation efficiency. ResultsA comprehensive evaluation of m⁶ATEpre was conducted through a series of experiments. We compared its performance against that of a similar prediction task model, as well as other classifiers. The results indicate that m⁶ATEpre achieved the best prediction performance. In addition, we analyzed different feature representation strategies and performed ablation experiments to validate the rationality of the model design. The results demonstrate that our proposed feature representation strategy has a greater advantage in improving prediction performance. In the HeLa cell line, bioinformatic analysis of the metagene distribution and sequence minimum free energy of m⁶A sites regulating translation efficiency (m⁶A-reg-TE sites) revealed their specific properties in translation regulation. Functional enrichment analysis indicated that m⁶A-reg-TE genes are associated with specific biological processes and KEGG pathways. By integrating the binding sites of YTHDF1 co-factors with m⁶A-reg-TE sites, we revealed that YTHDF1-mediated and m⁶A-dependent translation efficiency regulation requires the cooperation of multiple translation-regulatory RNA-binding proteins among its co-factors in the HeLa cell line. Furthermore, we extended our predictions to the dataset of the HEK293T cell line. Similarly, bioinformatic analysis of the metagene distribution and functional enrichment revealed the cell-specific characteristic of these predicted m⁶A-reg-TE sites in HEK293T cells. Likewise, integrated analysis of multiple YTHDF1 co-factors and m⁶A-reg-TE sites predicted in the HEK293T cell line reveals their m⁶A-dependent cooperation in regulating translation efficiency. Conclusionm⁶ATEpre is a timely tool that will advance our understanding of the mechanisms of m⁶A regulation in translation efficiency. The source code and datasets used in this work can be downloaded from https://www.scidb.cn/s/bAZZFr.

ObjectiveTo observe the clinical efficacy of Sanren Runchang formula in treating constipation-predominant irritable bowel syndrome （IBS-C） by regulating the brain-gut axis and the effects of the formula on serum levels of 5-hydroxytryptamine （5-HT）， vasoactive intestinal peptide （VIP）， and substance P （SP）. MethodsA randomized controlled design was adopted， and 72 IBS-C patients meeting Rome Ⅳ criteria were randomized into observation and control groups （36 cases）.The observation group received Sanren Runchang formula granules twice daily， and the control group received lactulose oral solution daily for 4 weeks. IBS Symptom Severity Scale （IBS-SSS）， IBS Quality of Life Scale （IBS-QOL）， and Bristol Stool Form Scale （BSFS） were used to assess clinical symptoms， and bowel movement frequency was recorded. The Self-Rating Anxiety Scale （SAS） and Self-Rating Depression Scale （SDS） were employed to evaluate psychological status. ELISA was employed to measure the serum levels of 5-HT， VIP， and SP. ResultsThe total response rate in the observation group was 91.67% （33/36）， which was higher than that （77.78%， 28/36） in the control group （χ²=4.50， P<0.05）. After treatment， both groups showed increased defecation frequency and BSFS scores， decreased IBS-SSS total score， abdominal pain and bloating scores， IBS-QOL health anxiety， anxiety， food avoidance， and behavioral disorders scores， SAS and SDS scores， serum 5-HT and VIP levels， and increased SP levels （P<0.05， P<0.01）. Moreover， the observation group showed more significant changes in the indicators above than the control group （P<0.05， P<0.01）. The SP level showed no significant difference between the two groups. During the 4-week follow-up， the recurrence rate was 5.88% in the observation group and 31.25% in the control group. No adverse events occurred in observation group， and 2 cases of mild diarrhea occurred in the control group. ConclusionSanren Runchang formula demonstrated definitive efficacy in alleviating gastrointestinal symptoms and improving the psychological status and quality of life in IBS-C patients， with a low recurrence rate. The formula can regulate serum levels of neurotransmitters such as 5-HT and VIP， suggesting its potential regulatory effect on the brain-gut axis through modulating neurotransmitters and neuropeptides. However， its complete mechanism of action requires further investigation through detection of additional brain-gut axis-related biomarkers.

ObjectiveTo construct a risk prediction model for frequent acute exacerbations of chronic obstructive pulmonary disease （COPD） under disease-syndrome combination， thus providing decision support for precise clinical intervention. MethodsA total of 2 029 patients with acute exacerbations of COPD admitted to the First Affiliated Hospital of Anhui University of Chinese Medicine from January 2020 to August 2024 were retrospectively included. These patients were classified into groups of frequent acute exacerbations （≥2 times/year） and infrequent acute exacerbations （<2 times/year） according to the hospitalization times per year. Risk factors were screened by LASSO regression combined with logistic regression， and a nomogram model was constructed. The model performance was assessed based on the area under the curve （AUC）， calibration curves， and decision curve analysis （DCA）. ResultsThe differences in baseline characteristics between the frequent acute exacerbations group （1 196 cases） and infrequent acute exacerbations group （833 cases） were not statistically significant. LASSO regression combined with multivariate logistic regression screened the following independent risk factors： body mass index （BMI）， hospitalization days， number of smoking years， place of residence， use of noninvasive ventilators， oxygen-demanding therapy， liver cirrhosis， use of systemic glucocorticosteroids， and traditional Chinese medicine syndrome （phlegm and stasis obstructing the lung）. The nomogram model showed good discrimination and calibration in both the training set （AUC=0.748） and validation set （AUC=0.774）. ConclusionThe risk prediction model for frequent acute exacerbations of COPD， integrating traditional Chinese medicine syndrome， constructed in this study has high accuracy. It can provide a scientific basis for early clinical identification of high-risk patients and individualized intervention.

This consensus focuses on the clinical application of domestic ultra-high-definition (UHD) medical endoscopes in cardiovascular surgery, addressing their clinical value, key technical aspects, and strategies for broader implementation. It traces the evolution of domestic endoscopic platforms from high-resolution imaging toward integrated advanced functionalities such as fluorescence imaging, three-dimensional visualization, and intelligent assistance. The document outlines standardized procedural steps and perioperative management pathways for minimally invasive endoscopic cardiovascular surgery and summarizes specific application scenarios and technical considerations in congenital heart disease, valvular disorders, coronary artery disease, arrhythmias, and cardiac tumors. Furthermore, recommendations are provided regarding professional training, multicenter collaboration, the establishment of clinical evaluation systems, and coordinated policy-industry support to promote standardized adoption and high-quality dissemination of domestic medical devices. Based on available evidence and expert agreement, 16 consensus statements are presented, aiming to offer authoritative and actionable guidance for clinical practice.

OBJECTIVE To investigate the molecular mechanism of Tiaozhong huashi decoction in improving gastroesophageal reflux disease （GERD） by regulating endoplasmic reticulum autophagy in rat esophageal interstitial cells of Cajal （ICC） through p62 signaling pathway.METHODS After establishing a rat GERD model via the weak acid intragastric administration and irregular diet， the rats were randomly divided into model group， western medicine group [1.56 mg/（kg·d） of mosapride， 3.125 mg/（kg·d） of lansoprazole] and low-， medium- and high-dose Tiaozhong huashi decoction [4.713， 9.425， 18.85 g/（kg·d）] groups. A blank group was also set up， with 8 rats in each group. The rats in each group were intragastrically administered with corresponding liquid or normal saline twice a day for 14 days. During the intervention period， the general conditions of rats’ hair， spirit， diet and defecation were observed. After the last gavage， the pathological changes of esophageal tissue and the ultrastructure of ICC were observed. The relative expression and positive expression levels of p62， autophagy-related protein 8 （ATG8）， 200 kDa FAK family kinase-interacting protein （FIP200） and Unc-51 like autophagy activating kinase 1 （ULK1） in esophageal tissue were detected.RESULTS Compared with the blank group， rats in the model group exhibited listlessness； thickening of the esophageal basal layer， elongation of lamina propria papillae with inflammatory cell infiltration； markedly dilated endoplasmic reticulum appearing as round vacuolar cisternae with ribosome detachment； and a small number of accumulated autophagosomes. The relative expression and positive expression levels of p62， ATG8， FIP200， and ULK1 were significantly decreased （ P ＜0.05）. Compared with the model group， the above pathological injuries in the medium- and high-dose Tiaozhong huashi decoction groups had all improved to varying degrees； the damaged endoplasmic reticulum structure was repaired， and no autophagosome accumulation was observed. In particular， the relative expression levels and positive expression levels of p62， ATG8， FIP200， and ULK1 in esophageal tissue of rats in the medium-dose Tiaozhong huashi decoction group were significantly increased （ P ＜0.05）.CONCLUSIONS Tiaozhong huashi decoction ameliorated esophageal mucosal injury in GERD rats， and its mechanism was possibly associated with activation of the p62 signaling pathway and restoration of the endoplasmic reticulum structure， morphology， and autophagic capacity of ICC.

Objective To explore the therapeutic effect of Chaizhu Tiaoshen Dingxuan Fang in the treatment of syndrome of stagnation of liver qi and spleen deficiency in vestibular migraine(VM).Methods A total of 72 VM patients admitted to Jilin Academy of Chinese Medicine Sciences from January to October 2024 were selected and divided into treatment group and control group according to the random number table method,with 36 cases in each group.Patients in control group were treated with flunarizine hydrochloride capsules,while patients in treatment group were treated with Chaizhu Tiaoshen Dingxuan Fang.Both groups of patients were treated continuously for 4 weeks.The therapeutic effect scores of TCM syndromes,visual analogue scale(VAS)scores of vertigo,dizziness handicap inventory(DHI)scores and hospital anxiety and depression scale(HADS)scores of two groups of patients were compared.Results After treatment,the therapeutic effect scores of TCM syndromes in both groups were significantly lower than those before treatment in the same group(P＜0.05),and the therapeutic effect scores of TCM syndromes in treatment group were significantly lower than those in control group(P＜0.05).After the treatment,VAS score of vertigo,HADS scores,DHI scores of each dimension and the total score in treatment group were significantly lower than those in before treatment and control group(P＜0.05).Conclusion Chaizhu Tiaoshen Dingxuan Fang can effectively relieve the dizziness symptoms of patients with syndrome of stagnation of liver qi and spleen deficiency in VM,and reduce their anxiety and depression.

Objective:To explore the cleaning quality of different cleaning methods on robotically luminal apparatuses.Methods:This research innovated and invented specially cleaning holder that could support boiling after reducing pressure,and protective device for preprocessing so as to better clean robotically luminal apparatuses.A total of 600 robotic luminal apparatuses,which were used in urology surgery at The First Affiliated Hospital with Nanjing Medical University in November 2023,were selected as the research object,and they were divided into control group,experimental group 1 and experimental group 2 according to the principle of randomization and balance,with 200 apparatuses in each group.The luminal apparatuses of control group were cleaned as the cleaning process of the manufacturer's instructions,and these of experimental group 1 were cleaned through ultrasonic cleaning machine with perfusion in the cleaning process of the manufacturer's instructions,and boiling after reducing pressure was selected to clean those of experimental group 2 through specially cleaning holder that was innovated and invented,which could support boiling after reducing pressure.Both two experiment groups adopted protective device that was invented by our research patent.The visual estimation and magnifying glass(5-10 times)with light source,adenosine triphosphate(ATP)fluorescence detector,and detection device for protein residue were adopted to test and judge cleaning quality,cleaning time,and wear rate of transportation in luminal apparatus after cleaning.Results:The qualified rates of visual estimation and magnifying glass with light source in control group,experiment group 1 and experiment group 2 were respectively 94.5%,99.0%and 99.5%,and the differences of pairwise comparison among three groups were significant(x2=6.44,8.591,P<0.05).The qualified rates of the fluorescence detection and the detection for protein residue in control group were respectively 93.5%and 90%,and those in experiment group 1 were respectively 98.5%and 97%,and the differences among two groups were significant(x2=6.51,11.41,P<0.05).For the cleaning time of single apparatus,the experiment group 1 was shortest,and the experiment group 2 was longest,and there were significant differences in pairwise comparison among three groups(t=2.981,48.178,34.419,P<0.05).The cleaning time of 8 cleaned apparatuses of experiment group 1 still can keep advantage,while there was not significant difference in that between it and control group(P>0.05).The cleaning time of 12 cleaned apparatuses of experimental group 2 was shortest,and that of experiment group 1 was longest,and the differences of that among three groups were significant(t=2.743,5.292,3.177,P<0.05).The average wear rates of transportation of control group,experiment group 1 and experiment group 2 were respectively(9.5±1.8)%,(6.2±1.5)%and(5.8±1.3)%,and the average wear rates of transportation of experiment group 1 and 2 were significantly lower than that of control group,and the differences of that in pairwise comparison among three groups were significant(t=21.7,24.3,P<0.05),while the difference of that between two experiment groups was not significant(P>0.05).Conclusion:The cleaning technique of boiling after reducing pressure can play better cleaning effect for the slender lumen of robotic apparatuses,while the wear rates of experiment group 1 and 2,which use protective device for preprocessing,are better than that of control group.The full load amount of the application of the special cleaning holder of the robotic luminal apparatuses of experiment group 2 can increase by 1 time,which is high efficient and convenient.

OBJECTIVE To investigate the differential expression of CXC chemokine receptor 1(CXCR1)in chronic rhinosinusitis(CRS)and its correlation with neutrophil infiltration.METHODS A total of 60 CRS patients with nasal polyps(CRSwNP group),30 CRS patients without nasal polyps(CRSsNP group),and 30 patients with deviated nasal septum(control group)were enrolled in this study.ELISA,immunohistochemistry,and reverse transcription polymerase chain reaction were employed to measure CXCR1 expression levels.Correlation analysis was conducted to evaluate the relationship between CXCR1 expression,neutrophil infiltration,and clinical symptom scores.RESULTS CXCR1 was highly expressed in both tissues and serum of CRSwNP patients.Serum CXCR1 levels showed positive correlations with peripheral blood neutrophil counts(r=0.363,P=0.004 4),neutrophil percentage(r=0.323,P=0.011 7),visual analog scale(VAS)score(r=0.328,P=0.010 5),Lund-Kennedy endoscopic score(r=0.331,P=0.009 9),and Lund-Mackay CT score(r=0.262,P=0.045 0).Tissue CXCR1 levels were positively correlated with tissue neutrophil counts(r=0.506,P=0.011 6)and percentage(r=0.564,P=0.004 1).CONCLUSION CXCR1 is highly expressed in CRSwNP patients,and its expression level is positively correlated with the degree of neutrophil infiltration and clinical symptom scores.Higher CXCR1 levels are associated with increased neutrophil migration in both serum and tissues,as well as more severe clinical symptoms.

Integrated metabolomic and lipidomic profiling,utilizing liquid chromatography coupled with high-resolution mass spectrometry(LC-HRMS),has emerged as a pivotal strategy for biomarker discovery.However,the inherent polarity disparity between metabolites and lipids complicates simultaneous analysis.To address this,a dual-stationary phase polarity-extended liquid chromatography(PELC)system was developed,which surpassed conventional one-dimensional LC(1D-LC)by enabling comprehensive coverage of both polar and non-polar compounds within a single injection.This system enhanced chromatographic resolution,peak capacity,and throughput while minimizing analytical variability.Extracellular vesicles(EVs),lipid bilayer-enclosed nanoparticles ubiquitously present in biofluids,had gained prominence as reservoirs of cancer biomarkers due to their cargo stability and pathophysiological relevance.Herein,the application of PELC-HRMS for concurrent metabolome-lipidome profiling in EVs was pioneered.A total of 193 metabolites were identified using this technique coupled with MS-DIAL software and Human Metabolome Database.Subsequently,this technique was employed to explore potential biomarkers for prostate cancer(PCa).Multivariate analysis identified 17 differentially abundant metabolites in PCa,implicating dysregulated pathways including purine metabolism,starch and sucrose metabolism,galactose metabolism,cysteine and methionine metabolism,and biosynthesis of unsaturated fatty acids.Notably,creatine(AUC=0.92)and DG 42:5(AUC=0.80)demonstrated robust diagnostic efficacy,attributable to their broad polarity ranges and EV-specific enrichment.This study established PELC as a high-fidelity platform for multi-omics integration in complex biospecimens,advancing mechanistic insights into metabolic rewiring and disease pathophysiology.