Objective: To establish a rapid, accurate, and decentralized workflow for bacterial whole-genome sequencing (WGS) and risk profiling within the shelf-life of platelet concentrates, and to characterize the species, virulence, antimicrobial resistance (AMR), and immune evasion mechanisms of co-existing bacteria in qualified platelet products, thereby providing a scientific basis for transfusion safety assessment. Methods: Three units of platelet concentrates, which tested negative by routine bacterial screening, were collected from the Chengdu Blood Center between May and June 2025. Samples were enriched at 37℃under six aerobic and nine anaerobic conditions for 7 days. Using a culturomics strategy, aliquots were plated for isolation on days 1, 3, 5, and 7 to obtain cultivable isolates, with negative culture controls included to exclude contamination. High-molecular-weight genomic DNA was extracted via mechanical grinding, purified, and size-selected. Sequencing libraries were constructed and sequenced on the G-seq500 single-molecule nanopore sequencing platform. Genomes were assembled using Flye and polished with NextPolish, with quality evaluated by BUSCO and CheckM. Taxonomic identification was performed using GTDB-Tk. Functional annotation and database comparisons were conducted to analyze virulence factors, AMR genes, and genes related to immune evasion and environmental adaptation. Results: Viable bacteria were successfully isolated from all three qualified platelet units within their shelf life. The isolates were identified as Bacillus albus, Niallia taxi, and Staphylococcus warneri. Nanopore sequencing generated 92 227-109 813 reads (totaling 680-758 Mb) with an N50 of 7 625-8 584 bp and Q20/Q30 scores of 97%/93%, respectively. All three genomes were assembled into complete circular chromosomes with 1-3 plasmids, achieving >93% completeness. Functional analysis revealed that B. albus carried multiple hemolysins, metalloproteases, and multidrug resistance genes, indicating the highest potential pathogenicity and AMR risk. S. warneri exhibited a typical multidrug resistance profile and regulatory network characteristic of coagulase-negative staphylococci, suggesting intermediate virulence. N. taxi harbored few canonical virulence factors and lacked functional AMR determinants, presenting a "low-virulence, low-resistance" profile. Notably, all three strains were enriched in genes encoding antimicrobial peptide resistance systems (e.g., dltABCD, mprF, GraRS, BceAB) and antioxidant enzymes, suggesting a strong capacity to withstand immune stress in the blood environment. Conclusion: Viable bacteria can be recovered from qualified platelet concentrates that test negative by routine screening. Nanopore WGS enables rapid strain-level identification and comprehensive risk profiling of virulence, resistance, and immune adaptation traits. The functional repertoires of these "co-existing" isolates range from environmental adaptation to potential pathogenicity, representing an underappreciated risk for transfusion-transmitted infections in susceptible recipients.

Objective:To evaluate respiratory muscle function before and after respiratory function training using surface electromyography(sEMG)in stroke patients with hemiplegia during recovery period.Method:Forty stroke patients with hemiplegia in the recovery phase were randomly divided into an observa-tion group and a control group,with 20 patients in each group.The control group was given routine rehabilita-tion treatment,while the observation group was given routine rehabilitation treatment combined with respirato-ry function training.Root Mean Square value(RMS)of respiratory muscle surface electromyography,pulmo-nary function indicators such as forced vital capacity(FVC),the forced expiratory volume in 1 second(FEV1),peak expiratory flow rate(PEF),and Fugl-Meyer Motor Scale(FMMS),Berg Balance Scale(BBS)were evaluated before and after 4 weeks of treatment in both groups.Result:After 4 weeks of treatment,the RMS of bilateral sternocleidomastoid muscles,parasternal external in-tercostal muscles,and diaphragmatic muscles,pulmonary function indicators including FVC,FEV1,PEF,FMMS and BBS in both groups were significantly improved(P＜0.05),and the scores of all indicators in the observation group were higher than those in the control group(P＜0.05).The correlation analysis showed that there was a certain correlation between the RMS of respiration muscles and indicators of pulmonary function,FMMS,BBS respectively in two groups after treatment(P＜0.05);the correlation between the RMS of respira-tion muscles and indicators of pulmonary function,FMMS,BBS in the observation group(r＞0.5,P＜0.05)was higher than that in the control group(r＜0.5,P＜0.05).Conclusion:Respiratory function training can significantly improve pulmonary function in stroke patients with hemiplegia during recovery period.Surface electromyography can be used to evaluate the respiratory muscle contraction function in stroke patients with hemiplegia.

Objective To investigate the efficacy of voriconazole in the treatment of pulmonary tuberculosis complicated with chronic pulmonary aspergillosis(CPA)based on CYP2C19 gene polymorphism detection and examine the factors affecting the efficacy for improving targeted therapy in clinical practice.Methods A total of 207 patients with pulmonary tuberculosis complicated with CPA treated in the Third People's Hospital of Kunming from December 2018 to November 2022 were randomly assigned to an observation group(105 cases)or a control group(102 cases).The patients in the control group received standard voriconazole treatment,while the patients in the observation group had their voriconazole regimen tailored based on CYP2C19 genotyping results.Plasma drug concentration levels,efficacy,and safety were compared between the two groups and in terms of CYP2C19 genotypes.Logistic regression analysis was used to identify the factors affecting treatment efficacy.Results The observation group showed significantly higher plasma voriconazole concentrations and overall antifungal efficacy compared to the control group(P＜0.05).In the observation group,CYP2C19 genotyping identified 37 extensive metabolizers(EM),47 intermediate metabolizers(IM),and 21 poor metabolizers(PM).Plasma concentration of voriconazole did not show significant difference between EM and IM(P＞0.05),but both PM and IM were associated with significantly lower plasma concentration of voriconazole than PM(P＜0.05).The clinical efficacy rate was 100％for PM,91.5％for IM,and 83.8％for EM(P＜0.05).The incidence of adverse events did not show significant difference among the three genotypes(P＞0.05).Logistic regression analysis revealed that lung cavitation,hypoalbuminemia,and agranulosis were significantly correlated with therapeutic efficacy(P＜0.05).Conclusions CYP2C19 gene polymorphism detection is valuable in clinical practice.It can inform anti-aspergillus therapy with voriconazole to effectively improve symptoms and clinical efficacy in patients with pulmonary tuberculosis complicated with CPA.Meanwhile,clinicians should be aware of the factors such as hypoproteinemia,agranulocytosis,and lung cavitation that may affect the efficacy of voriconazole.

Objective To compare the anti fungal efficacy and safety of amphotericin B cholesteryl sulfate complex(amphotericin B colloidal dispersion,ABCD)and amphotericin B for injection(amphotericin B deoxycholate,AmB-D)in the treatment of AIDS complicated with talaromycosis(TSM).Methods A total of 80 patients who were diagnosed with AIDS and complicated with TSM from December 2021 to January 2024 in Department of Infection,Kunming Third People's Hospital were included in the study.The patients were randomized to receive ABCD(n=40)via intravenous infusion or AmB-D(control,n=40)via Ⅳ infusion protected from light.The overall treatment efficacy rate,CD4+T lymphocyte count,routine blood tests,liver and kidney function tests,K+concentration,and the incidence of adverse drug reactions(ADR)during study were compared between the two treatment groups.Results The overall efficacy rate was 87.5％(35/40)in ABCD group and 80.0％(32/40)in the control(AmB-D)group(P＞0.05).WBC,hemoglobin,and platelet count were significantly higher after treatment compared with pretreatment levels(P＜0.05)in both groups.The CD4+T lymphocyte count was higher after treatment compared with pretreatment levels in both groups.And the CD4+T lymphocyte count in ABCD group was significantly higher than that in the control group(P＜0.05).The levels of total bilirubin,aspartate aminotransferase,alanine aminotransferase,blood urea nitrogen,and serum creatinine increased after treatment in both groups.Blood urea nitrogen and serum creatinine increased significantly in control group compared with ABCD group(P＜0.05).After treatment,serum K+concentration decreased significantly in control group compared with the pretreatment level and compared with ABCD group(P＜0.05).The incidence of adverse events in ABCD group was significantly lower than that in the control group.The time to renal injury was delayed significantly(P＜0.05).Conclusions In the treatment of AIDS complicated with TSM,the efficacy of ABCD was comparable to AmB-D.The incidence of hepatic impairment did not show significant difference between ABCD and AmB-D.However,ABCD is associated with less renal impairment,lower incidence of adverse events,and better safety,which is valuable for clinical use.

Objective To investigate the influencing factors of antibiotic-associated diarrhea(AAD)following congenital heart disease(CHD)surgery in pediatric patients,develop a nomogram-based predictive model,and validate its efficacy.Methods A retrospective analysis was conducted on the clinical data of pediatric patients who underwent CHD surgery in the Pediatric Intensive Care Unit(PICU)of a tertiary hospital in Guang-dong Province from July 2022 to July 2024.Patients were categorized into an AAD group and a non-AAD group.Univariate and multivariate logistic regression analyses were performed to identify risk factors for AAD occurrence following CHD surgery.A risk prediction model was developed,and a nomogram was constructed.The predictive performance of the model was evaluated using the Receiver Operating Characteristic(ROC)curve to calculate the area under the curve(AUC),the Hosmer-Lemeshow goodness-of-fit test,calibration curves,and clinical decision curve analysis.External validation of the model was conducted using data from patients in the Surgical Intensive Care Unit(SICU).Results The incidence of AAD following CHD surgery was 48.52%(229 out of 472 cases).Risk factors for AAD included the combined use of antibiotics,mechanical ventilation,elevated C-reactive protein levels,prolonged surgical duration,and extended antibiotic usage time(all with OR>1,P<0.05).Conversely,probiotic administration was identified as a protective factor(OR<1,P<0.05).The predictive model demon-strated excellent discrimination,as evidenced by the ROC curve areas:0.922(95%CI:0.894～0.951)in the modeling group,0.886(95%CI:0.838～0.915)in the internal validation group,and 0.862(95%CI:0.784～0.941)in the external validation group.Additionally,the model exhibited satisfactory calibration,as indicated by the Hosmer-Lemeshow test results:χ2=7.96,P=0.538 in the modeling group;χ2=4.24,P=0.895 in the inter-nal validation group;and χ2=9.923,P=0.270 in the external validation group.Furthermore,the model provided significant clinical utility.Conclusions Combined antibiotic use,duration of antibiotic therapy,mechanical ventilation,surgical duration,C-reactive protein(CRP)levels,and probiotic administration are key factors influ-encing the occurrence of AAD.The risk prediction model developed based on these variables demonstrates robust predictive performance and can serve as a valuable reference for the development and implementation of preventive and therapeutic strategies in clinical practice.

Objective:To evaluate respiratory muscle function before and after respiratory function training using surface electromyography(sEMG)in stroke patients with hemiplegia during recovery period.Method:Forty stroke patients with hemiplegia in the recovery phase were randomly divided into an observa-tion group and a control group,with 20 patients in each group.The control group was given routine rehabilita-tion treatment,while the observation group was given routine rehabilitation treatment combined with respirato-ry function training.Root Mean Square value(RMS)of respiratory muscle surface electromyography,pulmo-nary function indicators such as forced vital capacity(FVC),the forced expiratory volume in 1 second(FEV1),peak expiratory flow rate(PEF),and Fugl-Meyer Motor Scale(FMMS),Berg Balance Scale(BBS)were evaluated before and after 4 weeks of treatment in both groups.Result:After 4 weeks of treatment,the RMS of bilateral sternocleidomastoid muscles,parasternal external in-tercostal muscles,and diaphragmatic muscles,pulmonary function indicators including FVC,FEV1,PEF,FMMS and BBS in both groups were significantly improved(P＜0.05),and the scores of all indicators in the observation group were higher than those in the control group(P＜0.05).The correlation analysis showed that there was a certain correlation between the RMS of respiration muscles and indicators of pulmonary function,FMMS,BBS respectively in two groups after treatment(P＜0.05);the correlation between the RMS of respira-tion muscles and indicators of pulmonary function,FMMS,BBS in the observation group(r＞0.5,P＜0.05)was higher than that in the control group(r＜0.5,P＜0.05).Conclusion:Respiratory function training can significantly improve pulmonary function in stroke patients with hemiplegia during recovery period.Surface electromyography can be used to evaluate the respiratory muscle contraction function in stroke patients with hemiplegia.

Objective:To explore effect and potential mechanism of disulfiram on necrotizing apoptosis of renal podocytes and macrophages.Methods:Mouse renal podocyte MPC-5 and macrophages J774A.1 and BMDM cells were cultured in vitro and treated with TNF-α,Smac mimetic LCL-161 and pan-caspase inhibitor IDN-6556(TSI)to induce necroptosis.Cell necrosis was detected by propi-dium iodide staining.Western blot was used to detect protein levels of necroptosis markers MLKL,RIPK3 and RIPK1.Immuno-fluorescence microscopy was used to observe the subcellular distribution of RIPK3 and p-MLKL in MPC-5 cells.Results:TSI treat-ment induced significant necroptosis in both MPC-5 cells and macrophages.Disulfiram was able to inhibit necroptosis in these cells in a dose-dependent manner.Moreover,disulfiram markedly blocked the phosphorylation of RIPK1,RIPK3 and MLKL.The aggregation of RIPK3 and p-MLKL were also suppressed by disulfiram.Conclusion:Disulfiram inhibits necroptosis in podocytes and macrophages by suppressing RIPK1/RIPK3/MLKL signaling pathway.

Objective:To analyze the diagnostic value of bedside capsule endoscopy in patients with acute or severe gastrointestinal bleeding.Methods:Clinical data from patients who underwent bedside capsule endoscopy due to acute or severe suspected gastrointestinal bleeding in Nanfang Hospital, Southern Medical University from June 2018 to September 2021 were analyzed retrospectively. The efficacy of capsule endoscopy in detecting upper gastrointestinal tract and small intestinal bleeding was evaluated.Results:A total of 74 patients underwent bedside capsule endoscopy for suspected acute or severe gastrointestinal bleeding. Five patients were excluded due to failure of examination due to retention of capsule endoscope in the gastric lumen, and 69 were included in the study, of whom 54 patients with a definitive diagnosis of gastrointestinal hemorrhage. The positive detection rate of the capsule endoscopy was 83.33% (45/54), including 17 cases of ulcer, 5 cases of erosion, 5 cases of vascular malformation, 4 protrusion mass, 4 diverticulum, 5 obscure gastrointestinal bleeding, 1 stenosis , 1 active mucosal blood exudation, 1 gastric retention, 1 mucosal swelling, and 1 mucosal wrinkle change. The sensitivity and specificity of capsule endoscopy in the diagnosis of upper gastrointestinal bleeding were 92.31% (12/13) and 75.00% (3/4) respectively. The sensitivity and specificity of capsule endoscopy for diagnosing small intestinal bleeding were 80.49% (33/41) and 90.91% (10/11) respectively.Conclusion:Bedside capsule endoscopy demonstrates high sensitivity and specificity in the diagnosis of gastrointestinal bleeding, showing potential advantages in bedside applications for acute and severe gastrointestinal bleeding.

Objective To investigate the influencing factors of antibiotic-associated diarrhea(AAD)following congenital heart disease(CHD)surgery in pediatric patients,develop a nomogram-based predictive model,and validate its efficacy.Methods A retrospective analysis was conducted on the clinical data of pediatric patients who underwent CHD surgery in the Pediatric Intensive Care Unit(PICU)of a tertiary hospital in Guang-dong Province from July 2022 to July 2024.Patients were categorized into an AAD group and a non-AAD group.Univariate and multivariate logistic regression analyses were performed to identify risk factors for AAD occurrence following CHD surgery.A risk prediction model was developed,and a nomogram was constructed.The predictive performance of the model was evaluated using the Receiver Operating Characteristic(ROC)curve to calculate the area under the curve(AUC),the Hosmer-Lemeshow goodness-of-fit test,calibration curves,and clinical decision curve analysis.External validation of the model was conducted using data from patients in the Surgical Intensive Care Unit(SICU).Results The incidence of AAD following CHD surgery was 48.52%(229 out of 472 cases).Risk factors for AAD included the combined use of antibiotics,mechanical ventilation,elevated C-reactive protein levels,prolonged surgical duration,and extended antibiotic usage time(all with OR>1,P<0.05).Conversely,probiotic administration was identified as a protective factor(OR<1,P<0.05).The predictive model demon-strated excellent discrimination,as evidenced by the ROC curve areas:0.922(95%CI:0.894～0.951)in the modeling group,0.886(95%CI:0.838～0.915)in the internal validation group,and 0.862(95%CI:0.784～0.941)in the external validation group.Additionally,the model exhibited satisfactory calibration,as indicated by the Hosmer-Lemeshow test results:χ2=7.96,P=0.538 in the modeling group;χ2=4.24,P=0.895 in the inter-nal validation group;and χ2=9.923,P=0.270 in the external validation group.Furthermore,the model provided significant clinical utility.Conclusions Combined antibiotic use,duration of antibiotic therapy,mechanical ventilation,surgical duration,C-reactive protein(CRP)levels,and probiotic administration are key factors influ-encing the occurrence of AAD.The risk prediction model developed based on these variables demonstrates robust predictive performance and can serve as a valuable reference for the development and implementation of preventive and therapeutic strategies in clinical practice.

Objective To investigate the efficacy of voriconazole in the treatment of pulmonary tuberculosis complicated with chronic pulmonary aspergillosis(CPA)based on CYP2C19 gene polymorphism detection and examine the factors affecting the efficacy for improving targeted therapy in clinical practice.Methods A total of 207 patients with pulmonary tuberculosis complicated with CPA treated in the Third People's Hospital of Kunming from December 2018 to November 2022 were randomly assigned to an observation group(105 cases)or a control group(102 cases).The patients in the control group received standard voriconazole treatment,while the patients in the observation group had their voriconazole regimen tailored based on CYP2C19 genotyping results.Plasma drug concentration levels,efficacy,and safety were compared between the two groups and in terms of CYP2C19 genotypes.Logistic regression analysis was used to identify the factors affecting treatment efficacy.Results The observation group showed significantly higher plasma voriconazole concentrations and overall antifungal efficacy compared to the control group(P＜0.05).In the observation group,CYP2C19 genotyping identified 37 extensive metabolizers(EM),47 intermediate metabolizers(IM),and 21 poor metabolizers(PM).Plasma concentration of voriconazole did not show significant difference between EM and IM(P＞0.05),but both PM and IM were associated with significantly lower plasma concentration of voriconazole than PM(P＜0.05).The clinical efficacy rate was 100％for PM,91.5％for IM,and 83.8％for EM(P＜0.05).The incidence of adverse events did not show significant difference among the three genotypes(P＞0.05).Logistic regression analysis revealed that lung cavitation,hypoalbuminemia,and agranulosis were significantly correlated with therapeutic efficacy(P＜0.05).Conclusions CYP2C19 gene polymorphism detection is valuable in clinical practice.It can inform anti-aspergillus therapy with voriconazole to effectively improve symptoms and clinical efficacy in patients with pulmonary tuberculosis complicated with CPA.Meanwhile,clinicians should be aware of the factors such as hypoproteinemia,agranulocytosis,and lung cavitation that may affect the efficacy of voriconazole.