Objective: To validate the performance of the Procleix UltrioPlex E assay (Grifols, Spain) on the Procleix Panther automated nucleic acid detection platform, which employs the TMA method to simultaneously detect HIV-1/HIV-2/HCV/HBV/HEV viruses, and to evaluate its value for screening transfusion-associated infectious diseases. Methods: In accordance with the requirements of ISO15189"Application of the Guidelines for the Accreditation of Quality and Capabilities of Medical Laboratories in the Field of Molecular Diagnostics (CNAS-CL02-A009: 2018)", "Guidelines for Performance Validation of Molecular Diagnostic Testing Procedures (CNAS-GL039: 2019)", and the "Technical Operating Procedures for Blood Banks (2019 Edition)", this study validated the reagent's performance in terms of analytical sensitivity validation, performance consistency validation, interference resistance, and cross-contamination resistance. Results: Probit analysis revealed that the 95% detection limits (95% confidence interval) for HBV, HCV, HIV, and HEV were 2.0 IU/mL, 1.5 IU/mL, 18.0 IU/mL and 3.7 IU/mL, respectively, which were consistent with the minimum detection limits stated in the kit's package insert and were comparable to the Procleix Ultrio Elite kit. Both kits were used to test the performance validation serum plate simultaneously, yielding results consistent with the serum plate (Kappa=1), indicating stable performance. Detection of medium-and low-concentration lipemia and weakly positive hemolysis samples demonstrated good interference resistance. Cross-contamination performance validation showed that the kit exhibited excellent cross-contamination resistance. Conclusion: The Procleix UltrioPlex E nucleic acid detection kit enables combined detection of HIV-1, HIV-2, HCV, HBV, and HEV, allowing single-test screening for multiple viruses in donor blood. The kit's analytical performance is stable and meets basic laboratory requirements, making it suitable for screening transfusion-associated infectious diseases in blood banks.

Tumor drug resistance is an important problem in the failure of chemotherapy and targeted drug therapy, which is a complex process involving chromatin remodeling. SWI/SNF is one of the most studied ATP-dependent chromatin remodeling complexes in tumorigenesis, which plays an important role in the coordination of chromatin structural stability, gene expression, and post-translation modification. However, its mechanism in tumor drug resistance has not been systematically combed. SWI/SNF can be divided into 3 types according to its subunit composition: BAF, PBAF, and ncBAF. These 3 subtypes all contain two mutually exclusive ATPase catalytic subunits (SMARCA2 or SMARCA4), core subunits (SMARCC1 and SMARCD1), and regulatory subunits (ARID1A, PBRM1, and ACTB, etc.), which can control gene expression by regulating chromatin structure. The change of SWI/SNF complex subunits is one of the important factors of tumor drug resistance and progress. SMARCA4 and ARID1A are the most widely studied subunits in tumor drug resistance. Low expression of SMARCA4 can lead to the deletion of the transcription inhibitor of the BCL2L1 gene in mantle cell lymphoma, which will result in transcription up-regulation and significant resistance to the combination therapy of ibrutinib and venetoclax. Low expression of SMARCA4 and high expression of SMARCA2 can activate the FGFR1-pERK1/2 signaling pathway in ovarian high-grade serous carcinoma cells, which induces the overexpression of anti-apoptosis gene BCL2 and results in carboplatin resistance. SMARCA4 deletion can up-regulate epithelial-mesenchymal transition (EMT) by activating YAP1 gene expression in triple-negative breast cancer. It can also reduce the expression of Ca²⁺ channel IP3R3 in ovarian and lung cancer, resulting in the transfer of Ca²⁺ needed to induce apoptosis from endoplasmic reticulum to mitochondria damage. Thus, these two tumors are resistant to cisplatin. It has been found that verteporfin can overcome the drug resistance induced by SMARCA4 deletion. However, this inhibitor has not been applied in clinical practice. Therefore, it is a promising research direction to develop SWI/SNF ATPase targeted drugs with high oral bioavailability to treat patients with tumor resistance induced by low expression or deletion of SMARCA4. ARID1A deletion can activate the expression of ANXA1 protein in HER2+ breast cancer cells or down-regulate the expression of progesterone receptor B protein in endometrial cancer cells. The drug resistance of these two tumor cells to trastuzumab or progesterone is induced by activating AKT pathway. ARID1A deletion in ovarian cancer can increase the expression of MRP2 protein and make it resistant to carboplatin and paclitaxel. ARID1A deletion also can up-regulate the phosphorylation levels of EGFR, ErbB2, and RAF1 oncogene proteins.The ErbB and VEGF pathway are activated and EMT is increased. As a result, lung adenocarcinoma is resistant to epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs). Although great progress has been made in the research on the mechanism of SWI/SNF complex inducing tumor drug resistance, most of the research is still at the protein level. It is necessary to comprehensively and deeply explore the detailed mechanism of drug resistance from gene, transcription, protein, and metabolite levels by using multi-omics techniques, which can provide sufficient theoretical basis for the diagnosis and treatment of poor tumor prognosis caused by mutation or abnormal expression of SWI/SNF subunits in clinical practice.

Background/Aims: Endoscopic radiofrequency ablation (ERFA) is a treatment option for superficial esophageal squamous cell neoplasia (ESCN), with a relatively low risk of stenosis; however, the long-term outcomes remain unclear. We aimed to compare the long-term outcomes of patients with widespread superficial ESCN who underwent endoscopic submucosal dissection (ESD) or ERFA. Methods: We retrospectively analyzed the clinical data of patients with superficial ESCN who underwent ESD or ERFA between January 2015 and December 2021. The primary outcome measure was recurrence-free survival. Results: Ninety-two and 33 patients with superficial ESCN underwent ESD and ERFA, respectively. The en bloc, R0, and curative resection rates for ESD were 100.0%, 90.2%, and 76.1%, respectively. At 12 months, the complete response rate was comparable between the two groups (94.6% vs 90.9%, p=0.748). During a median follow-up of 66 months, recurrence-free survival was significantly longer in the ESD group than in the ERFA group (p=0.004), while no significant differences in overall survival (p=0.845) and disease-specific survival (p=0.494) were observed.Preoperative diagnosis of intramucosal cancer (adjusted hazard ratio, 5.55; vs high-grade intraepithelial neoplasia) was an independent predictor of recurrence. Significantly fewer patients in the ERFA group experienced stenosis compare to ESD group (15.2% vs 38.0%, p=0.016). Conclusions The risk of recurrence was higher for ERFA than ESD for ESCN but overall survival was not affected. The risk of esophageal stenosis was significantly lower for patients who underwent ERFA.

Objective:To develop a deep learning model based on 3D super-resolution reconstruction technology and to analyze its feasibility and effectiveness in predicting paraglottic space invasion in hypopharyngeal cancer.Methods:A retrospective study was conducted involving 382 patients with hypopharyngeal squamous cell carcinoma treated at Qilu Hospital of Shandong University between January 2014 and December 2020. The cohort included 364 males and 18 females, with a mean age of 62±7 years. Patients were divided into a training set ( n=300) and a test set ( n=82) based on enrollment time. A generative adversarial network was used to perform 3D super-resolution reconstruction on contrast-enhanced CT images, improving spatial resolution by 16 times. A 2.5D deep learning strategy was employed to construct Resnet-NR and Resnet-SR models based on conventional and super-resolution images, respectively, to predict whether the paraglottic space was invaded. Model performance was evaluated using receiver operating characteristic (ROC) curves and area under the curve (AUC). A multi-reader multi-case study was conducted to assess the impact of the artificial intelligence (AI) model on clinicians′ diagnostic capabilities. Results:The super-resolution model Resnet-SR achieved the highest accuracy in both the training set (AUC=0.87, 95% CI: 0.84-0.90) and the test set (AUC=0.88, 95% CI: 0.81-0.96), significantly outperforming traditional clinical indicators (T stage, N stage, tumor diameter, and pathological differentiation degree) (AUC range: 0.55-0.70, all P<0.05). In comparison, the conventional-resolution model Resnet-NR achieved AUCs of 0.81 (95% CI: 0.77-0.84, P=0.005) and 0.80 (95% CI: 0.71-0.89, P=0.184) in the training and test sets, respectively. Using Resnet-SR to assist clinical decision-making improved the diagnostic accuracy of junior physicians (AUC=0.793 without AI assistance vs. AUC=0.871 with AI assistance, P=0.012) and significantly reduced diagnosis time for clinicians of all experience levels (86.5 s without AI assistance vs. 82.5 s with AI assistance, t=2.01, P=0.032). Conclusion:This study successfully develops a deep learning model based on 3D super-resolution reconstruction technology, which can assist in preoperative prediction of paraglottic space invasion in hypopharyngeal cancer. The AI-assisted tool improves diagnostic accuracy for junior physicians and enhances diagnostic efficiency for clinicians across all experience levels.

To summarize the nursing experience of a patient with coronary heart disease who was left in the radial artery during the removal of the radial artery sheath after coronary angiography via the radial artery pathway.Nursing points:to start the emergency transfer process,to shorten the treatment transfer time;to assist to locate the position of the sheath to provide a basis for the selection of surgical incision;to conduct dynamic assessment of hemostatic effect,prevention of radial artery occlusion;to closely monitor pain and signs to prevent vasovagal reflex;to implement the whole psychological intervention,and to reduce the psychological burden of patients and their families.The ruptured sheath tube was successfully removed by emergency surgery of vascular surgery.A total of 6 days after the operation,the patient was transferred to cardiac surgery for coronary artery bypass grafting,and was discharged 23 days later.After 3 months of follow-up,the blood supply of the limbs was good,and the incision healed well.

Objective:To analyze the correlation between thymus dose-volume parameters and lymphopenia in patients with early-stage breast cancer (BC) receiving adjuvant radiotherapy (RT).Methods:Medical records of 54 patients with early-stage BC who received postoperative adjuvant RT in the Second Affiliated Hospital of Soochow University from January to December 2019 were retrospectively analyzed. Absolute lymphocyte counts (ALC) were collected at 1 month before (baseline) and weekly during RT. Lymphopenia was graded based according to the common terminology criteria for adverse events version 5.0 and nadir/baseline ALC was calculated. The thymus was delineated according to anatomical boundaries in the original RT planning system. Dosimetric parameters were obtained from the dose volume histograms. Stepwise multiple linear regression analysis was used to explore the factors associated with nadir/baseline ALC. The cutoff values of dosimetric parameters for predicting ≥grade 3 lymphopenia were obtained using the receiver operating characteristic (ROC) curve.Results:The proportion of 54 patients experiencing ≥ grade 3 lymphopenia was 38.9%. The median value of thymus volume, mean dose, V 5 Gy, V 10 Gy were 14.02 cm 3, 4.95 Gy, 36.18%, and 6.61%, respectively. Stepwise multiple linear regression analysis revealed that baseline ALC ( P=0.005), quadrant location ( P=0.005) and mean thymus dose ( P<0.001) were significantly associated with nadir/baseline ALC. ROC curve analysis indicated that the cutoff values of thymus mean dose, V 5 Gy and V 10 Gy for predicting ≥ grade 3 lymphopenia were 6.12 Gy, 35.2%, and 7.4%, respectively. Conclusions:Lymphopenia in early-stage BC patients is significantly correlated with high dosimetric parameters of the thymus during postoperative adjuvant RT. Thymus may be considered as an organ at risk during RT.

Objective To explore the regulatory effect of methyltransferase-like 3(METTL3)on m6A methylation of Sigma-1 receptor(Sigma-1R)mRNA and mitophagy in cerebral microvascular endothelial cells of sepsis mice.Methods Twelve adult male C57BL/6J mice were randomly divided into the Sham group(without cecal ligation or puncture)and the sepsis group(received cecal ligation and puncture),with 6 mice in each group.m6A methylation levels,and relative expression of METTL3 and Sigma-1R mRNA and protein in cerebral micro-vascular tissues were detected by m6A Dot blot,RT-qPCR,and Western blot,respectively.The bEnd.3 cells were cultured in vitro and divided into the control group(normal culture),model group[lipopolysaccharide(LPS)-induced in vitro sepsis],si-NC group(LPS treat-ment combined with si-NC transfection),and si-METTL3 group(LPS treatment combined with si-METTL3 transfection).m6A Dot blot was used to detect m6A methylation levels of cells in each group;RT-qPCR was used to detect the mRNA expression of METTL3 and Sigma-1R;and Western blot was used to detect the expression of METTL3,Sigma-1R,and autophagy-related proteins of Bnip3,LC3B,and Beclin1.m6A chromatin immunoprecipitation-qPCR was used to detect m6A methylation level of Sigma-1R of cells in each group.JC-1 probe was used to detect the mitochondrial membrane potential,and the ATP assay kit was used to detect the mitochondrial ATP content.Results Compared with the Sham group,the sepsis group showed increased m6A methylation level and METTL3 expression(P<0.05),and decreased Sigma-1R expression in cerebral microvascular tissues(P<0.05).Compared with the control group,m6A methylation level,METTL3 expression and m6A methylation level of Sigma-1R mRNA increased,the expression of Sigma-1R,the normal rate and increase rate of mitochondrial membrane potential,and the ATP content decreased,and the protein expression of Bnip3,LC3B and Beclin1 downregulated in the model group and the si-NC group,with statistically significant differences(P<0.05).Compared with the si-NC group,m6A methylation level,METTL3 expression and m6A methylation level of Sigma-1R mRNA decreased,the expression of Sigma-1R,the normal rate and increase rate of mitochondrial membrane potential elevated,the ATP content increased,and the protein expression of Bnip3,LC3B and Beclin1 upregulated in the si-METTL3 group,with statistically significant differences(P<0.05).Conclusion Silencing METTL3 can inhibit m6A methylation of Sigma-1R mRNA and promote the mitophagy in cerebral microvascular endothelial cells of septic mice.

Objective To explore the association between phase angle(PhA)and sarcopenia in middle-aged and elderly patients with type 2 diabetes mellitus(T2DM),and to evaluate its predictive value for the risk of sarcopenia in these patients.Methods We collected data from 356 middle-aged and elderly T2DM patients hospitalized in the Department of Endocrinology,Nanjing Drum Tower Hospital Group Suqian Hospital from March 2022 to June 2024,including 274 patients with diabetes only and 82 patients with T2DM combined with sarcopenia.A Logistic regression analysis was conducted to assess the relationship between phase angle and sarcopenia.The predictive value of PhA for sarcopenia in T2DM patients was analyzed using the receiver operating characteristic(ROC)curve,and the trend of PhA with the severity of sarcopenia in T2DM patients was tested by the Jonckheere-Terpstra method.Results Univariate analysis showed that the PhA value in the T2DM with sarcopenia group was significantly lower than that in the diabetes alone group,with a statistically significant difference(P<0.05).Additionally,height,body mass,body mass index(BMI),waist circumference,arm circumference,calf circumference,fasting insulin,postprandial 2 h insulin,fasting C-peptide,postprandial 2 h C-peptide,triglycerides,albumin,blood urea nitrogen,body composition indicators,6 m walking speed,muscle mass,and muscle strength-related indicators were significantly lower in the T2DM with sarcopenia group compared to the diabetes alone group.Age,duration of diabetes,glycated hemoglobin,25-hydroxyvitamin D[25-(OH)D]were significantly higher in the T2DM with sarcopenia group,with statistically significant differences(P<0.05).Multivariate Logistic regression analysis indicated that,after adjusting for other factors,PhA remained associated with sarcopenia in T2DM patients(P<0.05),with a decreased PhA increasing the risk of sarcopenia.ROC curve analysis showed that the area under the curve(AUC)for PhA predicting sarcopenia in T2DM patients was 0.769(95% CI:0.710-0.829),indicating the predictive efficacy of PhA.Trend analysis demonstrated a significant negative correlation between PhA and the severity of sarcopenia in T2DM patients(P<0.05).Conclusion The PhA is significantly associated with sarcopenia in patients with T2DM.It can serve as an early predictive and diagnostic tool for sarcopenia in individuals with T2DM.

Objective To develop a postoperative precision nursing program for children with obstructive sleep apnea-hypopnea syndrome(OSAHS),so as to provide references for the improvement of nursing quality for children with OSAHS.Methods Literature in precision nursing for children with OSAHS was systematically reviewed and summarised based on precision nursing theory and a postoperative precision nursing program was drafted.Using the Delphi's Method,two rounds of expert consultation were conducted and the postoperative precision nursing program for children with OSAHS was revised and finalised.Results The response rates of the two rounds of expert consultation were 100.00%with the expert authority coefficient of 0.875.The Kendall coefficient of concordance(W)of levels 1,2 and 3 indices in the second round of expert consultation were 0.289,0.203,0.187,respectively(all P<0.01).The importance score of each inclicator ranged from 4.50～4.94.The coefficient of variations for the three-level indices ranged from 0.05 to 0.21.The finalised program composed 8 level-1 indices(postoperative position assessment,intervention,prevention and care of bleeding,maintaining postoperative airway patency,postoperative nausea and vomiting care,postoperative pain assessment and care,dietary care,complication prevention and care,discharge guidance),16 level-2 indices,and 22 level-3 indices.Conclusion The postoperative precision nursing program for the children with OSAHS is scientific,innovative and applicable.It provides clinical references for nursing of the children with OSAHS.

This study explored the effects and underlying mechanism of Raf kinase inhibitory protein(RKIP)on apoptosis in mast cells sensitized by Echinococcus granulosus cyst fluid.Bone marrow-derived mast cells(BMMCs)were isolated and cultured from RKIP knockout(KO)and wild-type(WT)C57BL/6 mice.Cells were divided into control and sensitized groups.The sensitized group was incubated for 24 h in RPMI1640 medium containing 10%serum from mice infected with E.granulosus,then activated for 3 h or 6 h with E.granulosus cyst fluid.The control group was incubated for 24 h in RPMI1640 medium,and then received an equal vol-ume of PBS.Cells and supernatants were collected for analysis.Flow cytometry was used to detect the expression of CD117 and FcεRⅠα on BMMCs.The levels of β-hexosaminidase,IL-4,and TNF-α in the supernatant were quantified with ELISA.Western blot analy-sis was used to assess expression changes in RKIP,apoptosis-related proteins,and pathway proteins in BMMC before and after sensi-tization.Flow cytometry analysis revealed that after 4 weeks of induction,the CD117 and FcεRⅠα double-positivity rates on both WT and KO BMMC exceeded 90%.ELISA indicated that the E.granulosus cyst fluid resulted in significantly greater β-hexosaminidase re-lease(F=16.88,P<0.05),and levels of IL-4(F=16.51,P<0.05)and TNF-α(F=9.78,P<0.05)in the KO sensitized group than the WT sensitized group.With respect to the WT control group,the WT sensitized group showed significantly down-regulated pro-tein expression levels of RKIP(F=8.20,P<0.05)and Bcl-2(F=101.40,P<0.01)after 3 h,but significantly up-regulated levels of p-PI3K(F=8.04,P<0.05),p-Akt(F=32.52,P<0.01),p-P65(F=13.29,P<0.05),and cleaved-caspase-3(F=46.34,P<0.01).With respect to the WT sensitized group,the KO sensitized group showed significantly up-regulated protein expression of p-PI3K(F=8.45,P<0.05),p-Akt(F=8.58,P<0.05),p-P65(F=11.02,P<0.05),and Bcl-2(F=84.50,P<0.001)after 3 h,but significantly down-regulated expression of cleaved-caspase-3(F=15.66,P<0.05).In conclusion,RKIP may inhibit the PI3K/Akt/NF-κB pathway,thereby inducing apoptosis in mast cells sensitized by E.granulosus cyst fluid.This process may help ease aller-gic reactions caused by mast cells in echinococcosis,thus offering a promising new approach for preventing and treating such reactions.