To verify whether chaperones can promote the soluble expression of PlpE in Escherichia coli and whether the expressed protein is active,prokaryotic expression and Western blot detection were performed.The results showed that:The PlpE prokaryotic expression vector pET-32a(+)-plpE was expressed as inclusion body,and the expression form was not changed by changing the concentration of inducer,induction time and temperature.The companion proteins pG-KJE8,pGro7,pKJE7 and pG-Tf2 were co-expressed with pET-32a(+)-plpE in Eschierichia coli expres-sion system,respectively.When the final concentration of IPTG of 0.5 mmol/L,L-arabinose of 0.5 g/L or tetracycline of 5.0 μg/L were added as inducers and induced at 37 ℃ for 8 h,the results showed that the molecular companion pGro7 could change the expression of rp-PlpE from inclu-sion body to soluble expression.pG-KJE8,pKJE7 and pG-Tf2 had no effect on the expression of rp-PlpE.The soluble rp-PlpE can react specifically with the positive serum of Mannheimia haemolyti-ca.Therefore,the study showed that the co-expression of the chaperone protein pGro7 can make the rp-PlpE protein express in a soluble form,and the purified protein exhibits reactogenicity.These findings lay the foundation for the establishment of a subunit vaccine and serological diagno-sis methods for Mannheimia haemolytica.

Objective:To investigate the clinical value of 24-hour dynamic electrocardiogram combined with serum Annexin A1 (AnxA1) and endothelial microparticle (EMP) testing in the diagnosis of myocardial ischemia in patients with coronary heart disease without typical symptoms.Methods:A retrospective analysis was conducted on the clinical data of 80 patients with suspected coronary heart disease with myocardial ischemia and atypical symptoms who were admitted to The Third People's Hospital of Zhengzhou from January 2021 to April 2024. Based on the results of coronary angiography (CAG), patients were divided into groups: those with positive CAG results were assigned to the study group ( n = 49), while those with negative CAG results were assigned to the control group ( n = 31). The general clinical data, serum AnxA1 and EMP levels, duration of myocardial ischemia episodes, and the timing of ST-segment depression during myocardial ischemia were compared between the two groups. The efficacy, sensitivity, and specificity of each indicator for diagnosing myocardial ischemia in coronary heart disease without typical symptoms were analyzed using receiver operating characteristic curves. Results:In the study group, serum AnxA1 and EMP levels were (3.38 ± 1.02) μg/L and (1 415 ± 210) × 10 6/Ls, respectively, which were significantly higher than those in the control group, where the levels were (0.38 ± 0.11) μg/L and (1 179 ± 119) × 10 6/Ls ( t = 16.28, 5.70, both P < 0.05). Additionally, the duration of myocardial ischemia episodes in the study group was (275.17 ± 53.98) seconds, significantly longer than the (240.69 ± 44.58) seconds observed in the control group ( t = 2.97, P < 0.05). Receiver operating characteristic curve analysis results demonstrated the clinical value of the 24-hour dynamic electrocardiogram, serum AnxA1 and EMP levels, individually and in combination, for diagnosing myocardial ischemia in coronary heart disease without typical symptoms. Notably, the combined application showed the highest diagnostic efficacy, with an area under the curve of 0.961, a sensitivity of 0.980, and a specificity of 0.677. Conclusions:The use of 24-hour dynamic electrocardiogram and serum AnxA1 and EMP testing for the diagnosis of myocardial ischemia in coronary heart disease without typical symptoms all have certain clinical value, and the combined application shows even greater diagnostic effectiveness than a single application.

Objective To systematically evaluate qualitative studies on the experience of inhalation therapy for patients with chronic obstructive pulmonary disease(COPD),in order to provide a basis for healthcare professionals to optimize the care strategies.Methods A systematic search was conducted for qualitative studies on the experience of inhalation therapy for patients with COPD included in domestic and international databases,with a timeframe from the establishment of the database to September 2024.Literature quality was evaluated using the Australian JBI Centre for Evidence-Based Health Care Literature Quality Assessment Criteria for Qualitative Research,and the results were integrated by a pooled integration approach.Results A total of 12 articles were included,and 84 original findings were extracted and summarized into 10 new categories to form 4 integrated results,including low level of drug literacy,differences in perceived efficacy,multiple medication burdens,and multi-dimensional support for inhalation therapy.Conclusion COPD patients have multiple experiences during inhalation therapy.Medical staff should strengthen drug education,enhance patients'cognition and medication skills,foster a positive treatment attitude,and integrate multiple resources to improve patients'inhalation therapy experience.

OBJECTIVE To analyze the diagnosis of latent tuberculosis infection(LTBI)in hospitalized patients of a tertiary general hospital and investigate the current status of related comorbidities.METHODS The clinical data were collected from the 14 448 hospitalized patients who received tuberculin skin test(TST)or interferon-gamma release assay(IGRA)in China-Japan Friendship Hospital from Jan.1,2022 to Dec.31,2023,and the results were assessed.RESULTS The detection rate of LTBI was 23.62％(3413/14448)among the hospitalized patients who received the tests,and the rate of definite diagnosis was only 4.22％(144/3413).88.40％(3017/3413)of the hospitalized patients with LTBI had at least one type of comorbidity,and the top 5 comorbidities were in turn as follows:high blood pressure,hyperlipidemia,diabetes mellitus,malignant tumors and rheumatic immune disea-ses;the number of comorbidities was increased with the age(x2=291.199,P＜0.001).The rheumatic immune disease(73/144,50.69％)was the most common type of comorbidity among the hospitalized patients with definite diagnosis of LTBI,and less than half of the patients(66/144,45.83％)were treated in rheumatology and immu-nology department.CONCLUSION The two-way screening of LTBI and comorbidities is the core premise for the standardized management of LTBI.

BACKGROUND:Basement membrane genes are closely related to the occurrence and development of rheumatoid arthritis,but the role of basement membrane-related genes in the pathogenesis of rheumatoid arthritis under different traditional Chinese medicine patterns is not yet clear.OBJECTIVE:To explore the differences in the pathogenesis of rheumatoid arthritis with five different traditional Chinese medicine syndromes based on the analysis of basement membrane-related genes and transcriptomics,and to predict potential therapeutic drugs.METHODS:Rheumatoid arthritis-related traditional Chinese medicine syndrome microarray data and basement membrane-related genes were collected from the GEO database.The differentially expressed genes were screened using the R-limma package,and the expression trends were analyzed using the Mfuzz package.The protein-protein interaction network was constructed using the STRING database and key genes were selected using UPset.The differentially expressed genes were subjected to gene set enrichment analysis(GSEA)and enrichment analysis using the R-clusterProfiler package.Receiver operating characteristics curves were plotted to evaluate the diagnostic value of core basement membrane-related targets for each of the five syndromes.The immune infiltration of each syndrome was calculated using the CIBERSORT algorithm.Finally,potential traditional Chinese medicines and small molecule drugs targeting core basement membrane-related genes for the treatment of different traditional Chinese medicine syndromes of rheumatoid arthritis were predicted using SymMap and COREMINE databases.RESULTS AND CONCLUSION:(1)67,47,59,57,and 55 basement membrane-related differentially expressed genes were screened for the five traditional Chinese medicine syndromes of rheumatoid arthritis(obstruction syndrome,cold-dampness obstruction syndrome,liver-kidney deficiency syndrome,qi-blood deficiency syndrome,and blood stasis obstructing collaterals syndrome),with 5,7,5,3,and 5 key targets identified,respectively.(2)The most enriched biological processes in each syndrome were extracellular matrix adhesion,immune cell migration,collagen metabolism,and extracellular matrix receptor interaction,PI3K-Akt,focal adhesion,and Rap1 signaling pathways.(3)According to the predictions,Smilax glabra,Sargentodoxa cuneata,and Polygonatum sibiricum have the most potential as traditional Chinese medicines for the treatment of the five traditional Chinese medicine syndromes of rheumatoid arthritis by affecting basement membrane-related genes.(4)These results indicate that abnormal expression of basement membrane-related genes may affect the occurrence and development of rheumatoid arthritis through the regulation of cell adhesion,immune cell migration,and inflammatory reactions,among other pathways.These effects vary among different syndromes,with ITGA6 serving as a common diagnostic marker for the five traditional Chinese medicine syndromes of rheumatoid arthritis.Traditional Chinese medicines with heat-clearing and detoxifying properties may be potential effective drugs for the treatment of different syndromes of rheumatoid arthritis.

Objective To investigate the clinical application of ceftazidime/avibactam(CAZ/AVI)in lung trans-plant recipients with pulmonary infection caused by carbapenem-resistant Gram-negative bacilli(CRGNB),and ana-lyze the factors affecting the prognosis.Methods Lung transplant recipients who had CRGNB pulmonary infection and were treated with CAZ/AVI were included in the analysis.Based on 14-day clinical response,14-day microbial response,and 30-day survival status,the recipients were divided into a clinical response group and a clinical failure group,a microbial response group and a microbial failure group,as well as a survival group and a death group,re-spectively.Univariate analysis was conducted on various data from the two groups.Factors affecting therapeutic ef-ficacy and survival were included in a binary logistic regression model.Independent risk factors for CAZ/AVI anti-infective efficacy and all-cause mortality outcomes were analyzed.Results A total of 43 recipients were included.After 14-day anti-infective treatment,32 recipients(74.42％)achieved clinical response,and 30 recipients(69.77％)achieved microbial response.34 recipients(79.07％)survived 30 days after CAZ/AVI treatment.The Charlson comorbidity index(CCI),proportion of renal dysfunction,and incidence of shock in recipients in the clini-cal response group were all lower than those in the clinical failure group(P＜0.05),while the serum albumin(ALB)level was higher(P＜0.05).The incidence of shock in recipients in the microbial response group was lower than that in the microbial failure group(P＜0.05).CCI,proportion of renal dysfunction,and incidence of shock in recipients in the survival group were all lower than those in the death group(all P＜0.05),while ALB level was higher during treatment period(P＜0.05).Multivariate analysis of 14-day clinical response and 30-day survival showed that higher CCI was an independent risk factor affecting 14-day clinical response of recipients(OR=2.22,95％CI:1.07-4.63),while lower ALB levels(OR=0.72,95％CI:0.54-0.98)and higher CCI(OR=5.27,95％CI:1.18-23.58)were independent risk factors for 30-day all-cause mortality in recipients with pulmonary in-fection after lung transplant.Conclusion CAZ/AVI may be an effective drug for treating pulmonary infection caused by CRGNB in lung transplant recipients.Higher CCI is an independent risk factor for 14-day clinical failure in recipients after CAZ/AVI treatment.Lower ALB level and higher CCI are independent risk factors for increased 30-day mortality in recipients.

AIM:To investigate the impact of recombinant human CC16 protein(rhCC16)on cigarette smoke extract(CSE)-induced senescence-associated secretory phenotype(SASP)in human bronchial epithelial cells(HBECs)and in the lung tissues of chronic obstructive pulmonary disease(COPD)mice,and to explore the underlying mechanism.METHODS:HBECs were induced into cellular senescence using 5%CSE.The senescent HBECs were treated with 250 ng/mL rhCC16,and the levels of reactive oxygen species(ROS)were assessed using the 2',7'-dichlorodihydrofluorescein diacetate(DCFH-DA)method.The levels of trimethylated histone H3 at lysine 9(H3K9me3),a marker of senescence-associated heterochromatic foci(SAHF),were detected using a Western blot assay.RT-qPCR and ELISA were utilized to measure the mRNA expression and protein levels of SASP components including interleukin-1 beta(IL-1β),IL-6,IL-8,chemokine(C-X-C motif)ligand-1(CXCL-1),matrix metalloproteinase 1(MMP1)and MMP3.Passive smoking was con-ducted for six months to induce COPD in mice.RhCC16(2.5 μg/g body weight)or an equal volume of PBS(20 μL)was intranasally administered from the 16th week of smoking in the COPD+rhCC16 group or COPD+PBS group,respectively,with administration 2 hours before smoking.ROS levels in lung tissue cells were investigated using DCFH-DA staining.H3K9me3 levels in lung tissues were tested using Western blot assay.RT-qPCR and ELISA were performed to examine the mRNA expression and protein levels of IL-1β,IL-6,IL-8,CXCL-1,MMP1 and MMP3.RESULTS:DCFH-DA staining results showed that CSE stimulation increased ROS levels in HBECs,while rhCC16 treatment reduced them(P＜0.01).Western blot results indicated that CSE stimulation elevated H3K9me3 levels in HBECs,which were decreased with rhCC16 treatment(P＜0.01).RT-qPCR and ELISA assays demonstrated that CSE stimulation upregulated the mRNA and protein levels of IL-1β,IL-6,IL-8,CXCL-1,MMP1 and MMP3 in HBECs,which were reduced with rhCC16 admin-istration(P＜0.05).DCFH-DA staining results showed an increase in ROS levels in the lung tissues of COPD mice,which were decreased with rhCC16 administration(P＜0.01).Western blot data revealed an increase in H3K9me3 levels in the lung tissues of COPD mice,which were reduced with rhCC16 treatment(P＜0.01).RT-qPCR and ELISA assays demon-strated an upregulation of the mRNA and protein levels of IL-1β,IL-6,IL-8,CXCL-1,MMP1 and MMP3 in the lung tis-sues of COPD mice,which were reduced with rhCC16 treatment(P＜0.05).No statistically significant differences were ob-served in the above-mentioned indicators between the lung tissues of COPD and COPD+PBS mice(P＞0.05).CONCLU-SION:rhCC16 can effectively inhibit CSE-induced SASP in HBECs and in the lung tissues of COPD mice,with its under-lying mechanism potentially related to the inhibition of the ROS-H3K9me3 signaling pathway.

BACKGROUND:Basement membrane genes are closely related to the occurrence and development of rheumatoid arthritis,but the role of basement membrane-related genes in the pathogenesis of rheumatoid arthritis under different traditional Chinese medicine patterns is not yet clear.OBJECTIVE:To explore the differences in the pathogenesis of rheumatoid arthritis with five different traditional Chinese medicine syndromes based on the analysis of basement membrane-related genes and transcriptomics,and to predict potential therapeutic drugs.METHODS:Rheumatoid arthritis-related traditional Chinese medicine syndrome microarray data and basement membrane-related genes were collected from the GEO database.The differentially expressed genes were screened using the R-limma package,and the expression trends were analyzed using the Mfuzz package.The protein-protein interaction network was constructed using the STRING database and key genes were selected using UPset.The differentially expressed genes were subjected to gene set enrichment analysis(GSEA)and enrichment analysis using the R-clusterProfiler package.Receiver operating characteristics curves were plotted to evaluate the diagnostic value of core basement membrane-related targets for each of the five syndromes.The immune infiltration of each syndrome was calculated using the CIBERSORT algorithm.Finally,potential traditional Chinese medicines and small molecule drugs targeting core basement membrane-related genes for the treatment of different traditional Chinese medicine syndromes of rheumatoid arthritis were predicted using SymMap and COREMINE databases.RESULTS AND CONCLUSION:(1)67,47,59,57,and 55 basement membrane-related differentially expressed genes were screened for the five traditional Chinese medicine syndromes of rheumatoid arthritis(obstruction syndrome,cold-dampness obstruction syndrome,liver-kidney deficiency syndrome,qi-blood deficiency syndrome,and blood stasis obstructing collaterals syndrome),with 5,7,5,3,and 5 key targets identified,respectively.(2)The most enriched biological processes in each syndrome were extracellular matrix adhesion,immune cell migration,collagen metabolism,and extracellular matrix receptor interaction,PI3K-Akt,focal adhesion,and Rap1 signaling pathways.(3)According to the predictions,Smilax glabra,Sargentodoxa cuneata,and Polygonatum sibiricum have the most potential as traditional Chinese medicines for the treatment of the five traditional Chinese medicine syndromes of rheumatoid arthritis by affecting basement membrane-related genes.(4)These results indicate that abnormal expression of basement membrane-related genes may affect the occurrence and development of rheumatoid arthritis through the regulation of cell adhesion,immune cell migration,and inflammatory reactions,among other pathways.These effects vary among different syndromes,with ITGA6 serving as a common diagnostic marker for the five traditional Chinese medicine syndromes of rheumatoid arthritis.Traditional Chinese medicines with heat-clearing and detoxifying properties may be potential effective drugs for the treatment of different syndromes of rheumatoid arthritis.

Objective:To systematically evaluate the factors influencing the physical activity levels of patients with chronic obstructive pulmonary disease (COPD) .Methods:Computer retrieval of literature related to the influencing factors of physical activity in patients with COPD in PubMed, Embase, Web of Science, Cochrane Library, China National Knowledge Infrastructure, Wanfang, VIP Database, and China Biology Medicine disc, and the search period was from the establishment of the databases to March 2024. Two researchers independently screened the literature and conducted quality assessment according to inclusion and exclusion criteria. Meta-analysis was conducted using RevMan 5.4 software.Results:A total of 14 studies were included. The results of the Meta-analysis showed that gender [ OR=1.54, 95% CI (1.21, 1.96) ], degree of dyspnea [ OR=1.74, 95% CI (1.37, 2.21) ], fatigue [ OR=2.06, 95% CI (1.48, 2.87) ], and depression [ OR=1.63, 95% CI (1.30, 2.04) ] were influencing factors of physical activity levels in COPD patients ( P<0.05) . Conclusions:The physical activity levels of COPD patients are influenced by gender, degree of dyspnea, fatigue, and depression. Healthcare providers should develop personalized intervention goals and plans to improve patients' participation in and adherence to physical activity.

Objective:To retrieve, evaluate, and summarize the best evidence on ultrasound-guided placement and tip confirmation of peripherally inserted central catheter (PICC) in neonates.Methods:Based on the "6S" evidence pyramid model, literature on ultrasound-guided placement and tip confirmation of PICC in neonates was sequentially searched on guideline websites, professional society websites, and journal databases. The search period was from database establishment to September 30, 2024. Two researchers used uniform criteria for independent quality assessment and evidence extraction from the literature, and the extracted evidence was integrated and summarized.Results:A total of ten articles were included, including five guidelines, one clinical decision, two expert consensus, one Meta-analysis, and one evidence summary. Thirty pieces of evidence were developed in six aspects: personnel qualification and training, catheter selection, assessment of placement veins, ultrasound-guided PICC puncture in neonates, PICC tip confirmation in neonates, and prevention and management of malposition.Conclusions:This study summarizes the best evidence for ultrasound-guided placement and tip confirmation of PICC in neonates. It is recommended that evidence be selected and applied to develop a standardized process for neonatal PICC placement in conjunction with the resource environment of the department and the skill level of the healthcare professionals to improve the quality of nursing and to ensure the safe and effective use of the PICC in clinical practice.