Objective: To identify an optimal back-flush solution for leukocyte-depleted filters that maximizes peripheral blood mononuclear cell (PBMC) recovery with high viability, long-term storage stability, and sterility of the harvested residues, thereby providing a clinically translatable strategy. Methods: Three sterile bag-packaged solutions—Saline, Solvent, and Hanks' balanced salt solution (HBSS)—were used to back-flush randomly assigned leukocyte-depleted filters. Nucleated cell recovery rate and viability of the harvested residues were compared. The optimal solution identified was applied to an expanded sample set. PBMC viability and yield were evaluated after 1h vs 48h storage of the residues. PBMCs isolated from the residues were cryopreserved in liquid nitrogen for 1 month, followed by post-thaw comparisons of viability and T-cell expansion capacity. Results: The Solvent group achieved the highest and most consistent nucleated cell recovery rate. Post-flush recovery rate from filters after 400 mL whole blood processing was (21.3±1.6)% for the Solvent group, significantly higher than Saline group (19.2±6.3)% and HBSS group (11.2±5.0)%, with residues from all groups maintaining viability >90%. No biologically significant difference in residue viability was observed between 48h vs 1h storage groups (93.3±2.3)% vs (95.7±1.8)%). PBMC recovery rates from residues showed no statistical difference between 48h vs 1h storage groups [(48.2%±9.5%)vs (40.41%±8.35%), P>0.05], with (17.7±2.6)×10 cells. After 1-month cryopreservation and 10-day expansion, PBMCs isolated from 48-hour-stored residues retained (91.2±3.2)% viability and achieved a (61.9±15.9)-fold expansion. Conclusion: The bag-packaged Solvent, as a back-flush solution, enables sterile acquisition of leukocyte-depleted filter residues through closed-system tubing connections. These residues maintained PBMC viability and recovery rates after 48h storage at 2℃-8℃, with post-cryopreservation (1-month liquid nitrogen) viability and expansion capacity remaining stable. This protocol complies with blood bank regulatory criteria, addresses the concerns about the infectious window period in cell therapy raw materials, and provides a clinically translatable strategy for PBMC-based applications.

Objective: To investigate the effects of RING 1 and YY1 binding protein (RYBP), a member of the polycomb group (PcG), on bladder cancer cell proliferation and invasion, as well as the underlying mechanisms involved, so as to provide reference for the prevention and treatment of bladder cancer. Methods: Overexpressing and knocking down of RYBP were achieved in bladder cancer cell lines (T24, UM-UC-3, and 5637) via plasmids or siRNAs.Cell proliferation was assessed via thiazolyl blue (MTT) and colony formation assays, whereas migration was evaluated via scratch and Transwell assays.Changes in epithelial-mesenchymal transition (EMT) markers and other related proteins were examined with Western blotting. Results: RYBP overexpression significantly inhibited bladder cancer cell proliferation, invasion, and migration, whereas RYBP knockdown promoted these behaviors.Western blotting results revealed that RYBP overexpression downregulated the expressions of EMT markers N-cadherin, Vimentin, and Slug, but upregulated the expression of E-cadherin.Conversely, RYBP knockdown upregulated the expressions of N-cadherin, Vimentin, and Slug, while reducing the expression of E-cadherin. Conclusion: RYBP appears to inhibit the proliferation and migration of bladder cancer cells via the EMT pathway, indicating its potential application in bladder cancer therapies.

Objective:To analyze the electrocardiographic characteristics of patients with chronic pulmonary artery stenosis (PAS), and to explore their relationship with disease severity indicators.Methods:The study was a retrospective case-series analysis. Patients with chronic PAS admitted to Gansu Provincial Hospital from January 2018 to July 2021 were enrolled. The clinical data and the results of electrocardiography, transthoracic echocardiography, right cardiac catheterization, N-terminal B-type natriuretic peptide (NT-proBNP) measurement and 6-min walking distance test of patients were analyzed. The linear regression model or logistic regression model was used to analyze the relationship between electrocardiographic characteristics and the disease severity in patients with chronic PAS.Results:Sixty-three patients aged (62.1±9.7) years including 43 females (68.3%) were enrolled in the study. Among them, 62 patients (98.4%) had (R 1+S Ⅲ)-(S Ⅰ+R Ⅲ)<1.5 mV, and no patients had V 5lead R: S ratio to V 1 lead R: S<0.04 and V 6 lead R: S ratio<0.4. There were 55 patients (87.3%), with flat or inverted T-waves in V 1, and 10 patients (15.9%) with flat or inverted T-waves in all precordial leads (V 1-V 6). There were 18 patients (28.6%) with flat or inverted T-waves in inferior leads (Ⅱ, Ⅲ, aVF). Multiple liner regression analysis showed that Max R V1, 2+Max S I, aVL-S V1 combined with the number of flat or inverted T-waves in limb leads was independently correlated with atrial area ( R2=0.290, P=0.002); R V1+S V5 was independently correlated with right ventricular area ( R2=0.257, P=0.001); R peak V 1 combined with the number of flat or inverted T waves in precordial leads was independently correlated with tricuspid annular plane systolic excursion ( R2=0.407, P<0.001); (R 1+S Ⅲ)-(S Ⅰ+R Ⅲ) combined with the number of flat or inverted T waves in precordial leads was independently correlated with NT-proBNP ( R2=0.504, P<0.001); Max R V1, 2+Max S I, aVL-S V1 were independently correlated with right atrial pressure ( R2=0.803, P=0.036); (R 1+S Ⅲ)-(S Ⅰ+R Ⅲ) were independently correlated with mean pulmonary artery pressure ( R2=0.302, P<0.001); R aVRcombined with the number of flat or inverted T-waves in precordial leads was independently correlated with cardiac index ( R2=0.173, P=0.003); (R 1+S Ⅲ)-(S Ⅰ+R Ⅲ) was independently correlated with pulmonary vascular resistance ( R2=0.173, P=0.002); R peak V 1 combined with the number of flat or inverted T-waves in precordial leads was independently correlated with mixed vein oxygen saturation ( R2=0.302, P<0.001). Conclusion:The vast majority of patients with chronic PAS have (R 1+S Ⅲ)-(S Ⅰ+R Ⅲ)<1.5 mV and flat or inverted T-wave in V 1 lead, and some characteristic electrocardiographic manifestations are correlated with indicators of disease severity.

Objective To examine the influence of hormonal fluctuations on the perioperative outcomes of patients undergoing congenital heart surgery. Methods We conducted a retrospective analysis of clinical data from fertile women diagnosed with congenital heart disease at the Guangdong Provincial People's Hospital, between January 1, 2015, and July 30, 2019. Initially, patients were categorized into groups based on serum progesterone levels: a low progesterone group (n=31) and a high progesterone group (n=153). Furthermore, based on serum estrogen levels, they were divided into a low estrogen group (n=10), a medium estrogen group (n=32), and a high estrogen group (n=118) for comparative analysis. A control group (n=24) consisted of patients who received progesterone injections before their menstrual period. Results We finally included 184 patients. The patients’ average age was 27.6±5.7 years, with 142 (77.17%) presenting with complex congenital heart conditions. There were statistically significant differences in total postoperative standard thoracic drainage volume and postoperative albumin level between the high and low progesterone groups (P<0.05), while other perioperative outcome indicators showed no statistical differences (P>0.05). Among the different serum estrogen level groups, there were statistically significant differences in postoperative blood urea nitrogen levels, total postoperative standard thoracic drainage volume, and hospital stay (P<0.05), while other perioperative outcome indicators showed no statistical differences (P>0.05). Conclusion Considering the overall clinical significance, the physiological changes in sex hormone levels appear to have a negligible effect on the perioperative outcomes of fertile women with congenital heart disease.

Objective To investigate the expression levels and clinical significance of differentially expressed microRNAs(miRNAs)based on high-throughput sequencing in patients with left ventricular hypertrophy(LVH)and their possible regulatory mechanisms.Methods The miRNA sequencing was performed on plasma samples from 4 LVH patients and 4 healthy volunteers,and qRT-PCR validation was performed on plasma samples from 25 healthy volunteers and 35 LVH patients.The target genes of the obtained differentially expressed miRNA were predicted.The molecular function(MF),biological process(BP)and cellular components(CC)of these miRNAs were predicted by the Gene Ontology(GO)enrichment analysis,and their signal pathways were predicted by the Kyoto Encyclopedia of Genes and Genomes analysis.Meanwhile,the protein-protein interaction network was constructed.The diagnostic values of 2 differentially expressed miRNAs(hsa-miR-942-5p and hsa-miR-184)in LVH were analyzed by the area under the receiver operating characteristic(ROC)curve(AUCROC).Results A total of 945 differentially expressed miRNAs were identified,of which 1 and 9 were observed to be significantly upregulated and downregulated,respectively.The expression levels of hsa-miR-942-5p and hsa-miR-184 were verified by qRT-PCR to be significantly downregulated.The AUCROC of hsa-miR-942-5p and hsa-miR-184 were 0.694 6 and 0.880 4,respectively.Bioinformatics analysis revealed that the target genes were mainly enriched in the cell senescence,sphingolipid metabolism,and TGF-β,p53,and diabetic metabolism related signaling pathways.Conclusion The expression levels of hsa-miR-942-5p and hsa-miR-184 in plasma of LVH patients are significantly decreased,suggesting that they have good diagnostic potential.

Objective:To investigate the computed tomography (CT)-based integrated deep learning model for qualitative and quantitative classification of hepatic portal vein.Methods:The retrospective study was conducted. The CT imaging data of 291 patients undergoing upper-abdomen enhanced CT examination in the Beijing Tsinghua Changgung Hospital of Tsinghua University from October 2017 to January 2019 were collected. There were 195 males and 96 females, aged (51±12)years. The hepatic portal vein was reconstructed using the three-dimensional reconstruction system. Three-dimensional point cloud was input to the encoder model to obtain the three-dimen-sional reconstructed vectorized representation, which was used for qualitative classification and quantitative representation classification. Measurement data with normal distribution were repre-sented as Mean± SD, and comparison between groups was conducted using the paired t test. Count data were repre-sented as percentages or absolute numbers, and comparison between groups was analyzed using the paired chi-square test. Results:(1) Three-dimensional reconstruction of portal vein and anatomical classification. Three-dimensional structure was reconstructed in the 291 patients. Classification of main hepatic portal vein showed 211 cases of Akgul type A, 29 cases of Akgul type B, 16 cases of Akgul type C, 10 cases of Akgul type D, and 25 cases of unclassifiable. (2) Prediction of qualitative classification of main hepatic portal vein. Of the 291 patient samples, 25 unclassifiable or poor quality samples were excluded, 266 samples were used for automated qualitative classification of the main portal vein by machine model. There were 211 cases of Akgul type A, 29 cases of Akgul type B, 26 cases of Akgul type C&D. The Macro-F1 of 266 patients was 61.93%±40.50% and the accuracy was 84.99%, versus 32.38%±19.81% and 61.65% of Random classifier, showing significant differ-ences between them ( t=7.85, χ2=62.89, P<0.05). (3) Quantitative representation of portal vein classification. The probabilities of quantitative classification for Akgul qualitative classification of similar samples included P@1 as 73%±45%, P@3 as 70%±37%, P@5 as 69%±35%, P@10 as 67%± 32%, mean reciprocal rank(MRR) as 80%±34%, versus 57%±50%, 58%±35%, 58%±32%, 58%± 30%, 70%±37% of the baseline model, showing significant differences between the two analytical methods ( t=5.22, 5.11, 5.00, 4.99, 3.47, P<0.05). Conclusion:The automated classification model for the hepatic portal vein structure was constructed using CT-based three-dimensional reconstruc-tion and deep learning technology, which can achieve automatic qualitative classification and quanti-tatively describe the hepatic portal vein structure.

Objective This paper investigates the effect of a multi-physical field fusion intervention based on transcranial direct current stimulation(tDCS),transcranial magnetic stimulation(TMS)and music on mental fatigue.Methods After fatigue induction,10 subjects received blank group stimulation,music group stimulation and tDCS-TMS-Music stimulation,respectively.tDCS stimulation sites were located in the bilateral frontal regions of the subjects,and TMS stimulation sites were located in the bilateral occipital regions of the hindbrain.Heart rate variability and reaction performance were measured before and after each intervention to determine the elimination effect of different intervention programs on mental fatigue.Results Compared with the control group,the tDCS-TMS-Music group showed significantly greater improvements in subjective mental workload,response performance,and heart rate variability.Conclusion The results of this study support that tDCS-TMS-Music can effectively alleviate mental fatigue induced by long-term cognitive performance tasks,and the intervention effect is better than music intervention and resting-state relief at the same time.

Objective This study aimed to explore the expression trends of innate immune cells and immune-checkpoint molecules validated by data calculation in the process of oral mucosal carcinogenesis,as well as to explore methods of suppressing oral mucosal carcinogenesis based on immunotherapy by predicting their interactions.Me-thods 1)The cancer genome atlas(TCGA)database comprehensively scores immune cells and immune-checkpoint molecules in the process of oral mucosal carcinogenesis and screens out intrinsic immune cells and immune-check-point molecules that interfere with tumor immune escape.2)Clinical patient blood routine data were collected for the statistical analysis of peripheral blood immune cells during the progression of oral mucosal carcinogenesis.Immune cells in peripheral blood that may affect the progression of oral mucosal carcinogenesis were screened.3)Immunohis-tochemical staining was performed on intrinsic immune cells and immune-checkpoint molecules validated based on da-ta calculation in various stages of oral mucosal carcinogenesis.4)Special staining was used to identify innate immune cells in various stages of oral mucosal carcinogenesis based on data-calculation verification.5)Survival analysis was conducted on intrinsic immune cells and immune-checkpoint molecules validated based on data calculation during the process of oral mucosal carcinogenesis.The association of intrinsic immune cells and immune-checkpoint molecules with the prognosis of oral squamous cell carcinoma was verified.Results The expression of monocytes and neutro-phils increased during the process of oral mucosal carcinogenesis.The expression of eosinophils showed a single peak trend of up and down.The expression of mast cells decreased.In the process of oral mucosal carcinogenesis,the ex-pression of the immune-checkpoint molecules cytotoxic T-lymphocyte-associated protein 4(CTLA4)and programmed cell death-ligand(PD-L1)increased.The expression trends of monocytes,neutrophils,and eosinophils were positively correlated with those of CTLA4 and PD-L1 immune-checkpoint molecules.The expression trend of mast cells was negatively correlated with the expression of CTLA4 and PD-L1.Monocytes,neutrophils,and eosinophils may pro-mote tumor immune escape mediated by CTLA4 and/or PD-L1,thereby accelerating the progression of oral mucosal carcinogenesis.Mast cells may inhibit tumor immune escape mediated by CTLA4 and/or PD-L1,delaying the progres-sion of oral mucosal carcinogenesis.Conclusion Therefore,interference with specific immune cells in innate immu-nity can regulate the expression of CTLA4 and/or PD-L1 to a certain extent,inhibit tumor immune escape,and delay the progression of oral mucosal carcinogenesis.

Objective To establish interferon-induced transmembrane protein 3(Ifitm3)knockout mice and to explore the effects of Ifitm3 on the proliferation and differentiation of adult neural stem cells of mice(aNSCs).Methods IFITM3 knockout mice were established by the CRISPR/Cas9 method and identified by genotype identification and Western Blot.The differences between Ifitm3-knockout mice and wild-type mice were analyzed by hematoxylin-eosin(HE)staining and flow cytometry.The aNSCs of wild-type mice and Ifitm3-knockout mice were isolated and cultured,the number and size of neurospheres were detected,The ability of aNSCs to proliferate and differentiate were detected by quantitative reverse-transcription polymerase chain reaction,Western Blot,and immunofluorescence.Results Ifitm3-knockout mice were successfully established.The mice developed normally,and there were no obvious abnormalities either histopathologically or the immune system.In vitro experiments showed that Ifitm3 knockout inhibited the self-renewal potential of aNSCs,led to a decrease in the proliferation ability of aNSCs,and inhibited the differentiation of aNSCs into immature neurons and astrocytes.Conclusions This study finds that a lack of IFITM3 result in the ability of aNSCs to proliferate and differentiate decreased,IFITM3 may regulate the function of aNSCs.

Purpose To study the changes of white matter microstructure in moderate and severe obstructive sleep apnea-hypopnea syndrome(OSAHS)via synthetic MRI.Materials and Methods A total of 30 patients with moderate and severe OSAHS diagnosed for the first time and 30 healthy controls were prospectively collected in General Hospital of Taiyuan Iron&Steel(Group)Co.,LTD from October 2022 to September 2023.Both groups underwent routine MRI,3D-T1 MPRAGE and synthetic MRI scans.The T1,T2 and proton density(PD)values of multiple white matter regions in the two groups were compared,and the correlation between T1,T2 and PD values in different brain regions and sleep parameters was analyzed.Results Compared with the control group,the T1 value were increased in various white matter regions,including the right cerebral peduncle,cingulate fasciculus,posterior thalamic radiation,external capsule,bilateral internal capsule,corona radiata,superior longitudinal fasciculus and superior frontal-occipital fasciculus(t/Z=-3.267-3.286,all P<0.05).The T2 value of the left inferior cerebellar peduncle was decreased(Z=2.114,P=0.035),and the T2 value of the right posterior thalamic radiation and bilateral external capsule were increased(Z=-2.410,-2.484,-2.306,all P<0.05).Furthermore,the PD value were increased in corpus callosum,left internal capsule,sagittal layer,hippocampus,right posterior thalamic radiation,cingulate fasciculus,bilateral corona radiata and superior longitudinal fasciculus(t/Z=-3.726-2.757,all P<0.05).The apnea hypopnea index of patients with moderate and severe OSAHS was positively correlated with the T1 value(r=0.387,P=0.035)and PD value(r=0.382,P=0.037)of the right cingulate bundle,and there was no correlation between the lowest oxygen saturation and the quantitative parameters in moderate and severe OSAHS(P>0.05).Conclusion Synthetic MRI can sensitively observe the changes of white matter relaxation values in moderate and severe OSAHS,and has high application value in evaluating the damage of white matter in OSAHS.