Objective: To design and establish a new method for flow cytometry-based detection of commonly observed highly expressed antigens on red blood cells, and to further evaluate the differences and distribution characteristics of antigen expression levels between ABO blood type homozygotes and heterozygotes in healthy individuals. Methods: Residual blood samples after donor blood type identification by Shanghai Blood Center in April 2024 were collected. Among them, samples of 19 homozygous and 19 heterozygous individuals of type A and type B were selected. Then the expression level of ABO antigen on red blood cells were detected using the new method established in this study and the traditional aldehyde fixed red blood cell method. Both methods were tested independently three times and the results were compared. Results: The mean values of the three detection results of the new method was (×10 /RBC): AA homozygous 3.3±0.5, AO heterozygous 2.8±0.3, BB homozygous 3.6±0.3, BO heterozygous 3.1±2.8. The mean values of the three detection results of the aldehyde fixation method were AA homozygous 5.9±0.9, AO heterozygous 5.0±1.4, BB homozygous 3.8±0.6, and BO heterozygous 3.3±0.4. The average antigen distribution of each genotype followed a normal distribution. Comparing the average antigen expression levels of homozygotes and heterozygotes, both methods showed that A/B homozygotes had higher antigen levels than heterozygotes, with AA being 1.17 to 1.18 times that of AO and BB being 1.15 to 1.16 times that of BO. Comparing the inter batch differences in the three test results of two methods, the new method showed no significant difference in the three test results for four genotypes (P>0.05). The aldehyde fixation method showed significant differences in the test results for all three genotypes (P<0.01) except for BB homozygotes (P>0.05). The reliability and reproducibility of the new method were better than those of the traditional aldehyde fixation method. Conclusion: The antigen expression level of ABO homozygotes is higher than that of heterozygotes, and the difference in antigen level between type A homozygotes and heterozygotes is slightly higher than that of type B. The new method is superior to traditional aldolization fixation methods.

OBJECTIVE: Therapeutic angiogenesis has become a promising approach for treating ischemic heart disease (IHD). The present study aims to investigate the effects of Qishen Granule (QSG) on angiogenesis in myocardial ischemia (MI) and the potential mechanism. METHODS: In vivo study was conducted on rat model of myocardial infarction. QSG was performed daily at a dose of 2.352 g/kg for four weeks. Cardiac function was assessed by echocardiogram and pro-angiogenic effects were evaluated by Laser Doppler and CD31 expression. Oxygen-glucose deprivation (OGD) was applied in cultured human umbilical vein endothelial cells (HUVECs). Cell viability, wound healing and tube formation assay were used to test functions of HUVECs. ELISA and Western blots were used to assess protein expressions of bone morphogenetic protein 2-delta-like 4-notch homolog 1 (BMP2-Dll4-Notch1) signaling pathway. RESULTS: The results showed that QSG improved heart function, cardiac blood flow and microvessel density in myocardial ischemic rats. In vitro, QSG protected HUVECs by promoting the cell viability and tube formation. QSG upregulated bone morphogenetic protein-2 (BMP2) and downregulated delta-like 4 (Dll4) and notch homolog 1 (Notch1) expressions both in rats and HUVECs. CONCLUSION QSG protected against MI by promoting angiogenesis through BMP2-Dll4-Notch1 pathway. BMP2 might be a promising therapeutic target for IHD.

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Mechanical ventilation (MV) is currently widely used in the treatment of respiratory failure and anesthesia surgery, and is a commonly used respiratory support method for critically ill patients; however, improper usage of MV can lead to ventilator-induced lung injury (VILI), which poses a significant threat to patient life. Alveolar epithelial cell (AEC) has the functions of mechanosensation and mechanotransduction. Physiological mechanical stretching is beneficial for maintaining the lineage homeostasis and normal physiological functions of AEC cells, while excessive mechanical stretching can cause damage to AEC cells. Damage to AEC cells is an important aspect in the occurrence and development of VILI. Understanding the effects of mechanical stretching on AEC cells is crucial for developing safe and effective MV strategies, preventing the occurrence of VILI, and improving the clinical prognosis of VILI patients. From the perspective of cell mechanics, this paper aims to briefly elucidate the mechanical properties of AEC cells, mechanosensation and mechanotransduction of mechanical stretching in AEC cells, and the injury and repair of AEC cells under mechanical stretch stimulation, and potential mechanisms with the goal of helping clinical doctors better understand the pathophysiological mechanism of VILI caused by MV, improve their understanding of VILI, provide safer and more effective strategies for the use of clinical MV, and provide theoretical basis for the prevention and treatment of VILI.
Humans ; Mechanotransduction, Cellular ; Ventilator-Induced Lung Injury ; Stress, Mechanical ; Alveolar Epithelial Cells ; Respiration, Artificial/adverse effects* ; Epithelial Cells ; Pulmonary Alveoli/cytology* ; Animals

OBJECTIVE: To investigate the correlation between nucleated red blood cell (NRBC) level on the first day of intensive care unit (ICU) admission and 28-day mortality in adult septic patients, and to evaluate the value of NRBC as an independent predictor of death. METHODS: Single-cell transcriptomic analysis was performed using the GSE167363 dataset from the Gene Expression Omnibus (including 2 healthy controls, 3 surviving septic patients, and 2 non-surviving septic patients). A retrospective clinical analysis was conducted using the America Medical Information Mart for Intensive Care-IV (MIMIC-IV) database, including adult patients (≥ 18 years) with first-time admission who met the Sepsis-3.0 criteria, excluding those without NRBC testing on the first ICU day. The demographic information, vital signs, laboratory test indicators, disease severity score and survival data on the first day of admission were collected. The restricted cubic spline (RCS) curve was used to determine the optimal cut-off value of NRBC for predicting 28-day mortality in patients. Patients were divided into low-risk and high-risk groups based on this cut-off value for intergroup comparison, with Kaplan-Meier survival curve analysis conducted. Independent risk factors for 28-day mortality were analyzed using Logistic regression and Cox regression analysis, followed by the construction of regression models. RESULTS: NRBC were detected in the peripheral blood of septic patients by single-cell transcriptomic. A total of 1 291 sepsis patients were included in the clinical analysis, with 576 deaths within 28 days, corresponding to a 28-day mortality of 44.6%. RCS curve analysis showed a nonlinear relationship between the first-day NRBC level and the 28-day mortality. When NRBC ≥ 1%, the 28-day mortality of patients increased significantly. Compared to the low-risk group (NRBC < 1%), the high-risk group (NRBC ≥ 1%) had significantly higher respiratory rate, heart rate, sequential organ failure assessment (SOFA), and simplified acute physiology score II (SAPSII), and significantly lower hematocrit and platelet count. The high-risk group also had a significantly higher 28-day mortality [49.8% (410/824) vs. 35.5% (166/467), P < 0.05], and shorter median survival time (days: 29.8 vs. 208.6, P < 0.05). Kaplan-Meier survival curve showed that compared with the low-risk group, the survival time of high-risk group was significantly shortened (Log-rank test: χ ² = 25.1, P < 0.001). After adjusting for potential confounding factors including body mass, temperature, heart rate, respiratory rate, mean arterial pressure, serum creatinine, pulse oximetry saturation, hemoglobin, hematocrit, Na⁺, K⁺, platelet count, and SOFA score, multivariate regression analysis confirmed that NRBC ≥ 1% was an independent risk factor for 28-day mortality [Logistic regression: odds ratio (OR) = 1.464, 95% confidence interval (95%CI) was 1.126-1.902, P = 0.004; Cox regression: hazard ratio (HR) = 1.268, 95%CI was 1.050-1.531, P = 0.013]. CONCLUSIONS NRBC ≥ 1% on the first day of ICU admission is an independent risk factor for 28-day mortality in septic patients and can serve as a practical indicator for early prognostic assessment.
Humans ; Sepsis/blood* ; Intensive Care Units ; Risk Factors ; Retrospective Studies ; Prognosis ; Male ; Female ; Hospital Mortality ; Middle Aged ; Aged

Long non-coding RNAs(lncRNAs)play an indispensable role in the occurrence and development of ovarian cancer(OC).However,the potential involvement of lncRNAs in the progression of OC is largely unknown.To investigate the detailed roles and mechanisms of RAD51 homolog B-antisense 1(RAD51B-AS1),a novel lncRNA in OC,reverse transcription-quantitative polymerase chain reaction(RT-qPCR)was performed to verify the expression of RAD51B-AS1.Cellular proliferation,metastasis,and apoptosis were detected using the cell counting kit-8(CCK-8),colony-formation,transwell,and flow cytometry assays.Mouse xenograft models were established for the detection of tumorigenesis.The results revealed that RAD51B-AS1 was significantly upregulated in a highly metastatic human OC cell line and OC tissues.RAD51B-AS1 significantly increased the proliferation and metastasis of OC cells and enhanced their resistance to anoikis.Biogenetics prediction analysis revealed that the only target gene of RAD51B-AS1 was RAD51B.Subsequent gene function experiments revealed that RAD51B exerts the same biological effects as RAD51B-AS1.Rescue experiments demonstrated that the malignant biological behaviors promoted by RAD51B-AS1 overexpression were partially or completely reversed by RAD51B silencing in vitro and in vivo.Thus,RAD51B-AS1 promotes the malignant biological behaviors of OC and activates the protein kinase B(Akt)/B cell lymphoma protein-2(Bcl-2)signaling pathway,and these effects may be associated with the positive regulation of RAD51B expression.RAD51B-AS1 is expected to serve as a novel molecular biomarker for the diagnosis and prediction of poor prognosis in OC,and as a potential therapeutic target for disease management.

Objective:To identify diagnostic markers related to oxidative stress in chronic rhinosinusitis with nasal polyps (CRSwNP) by analyzing transcriptome sequencing data, and to investigate their roles in CRSwNP.Methods:Utilizing four CRSwNP sequencing datasets, differentially expressed genes (DEGs) analysis, weighted gene co-expression network analysis (WGCNA), and three machine learning methods for Hub gene selection were performed in this study. Subsequent validation was carried out using external datasets, as well as real-time quantitative polymerase chain reaction (Real-time qPCR), and immunofluorescence staining of clinical samples. Moreover, the diagnostic efficacy of the genes was assessed by receiver operating characteristic (ROC) curve, followed by functional and pathway enrichment analysis, immune-related analysis, and cell population localization. Additionally, a competing endogenous RNA (CeRNA) network was constructed to predict potential drug targets. Statistical analysis and plotting were conducted using SPSS 26.0 and Graphpad Prism9 software.Results:Through data analysis and clinical validation, CP, SERPINF1 and GSTO2 were identified among 4 138 DEGs as oxidative stress markers related to CRSwNP. Specifically, the expression of CP and SERPINF1 increased in CRSwNP, whereas that of GSTO2 decreased, with statistically significant differences ( P<0.05). Additionally, an area under the curve (AUC)>0.7 indicated their effectiveness as diagnostic indicators. Importantly, functional analysis indicated that these genes were mainly related to lipid metabolism, cell adhesion migration, and immunity. Single-cell data analysis revealed that SERPINF1 was mainly distributed in epithelial cells, stromal cells, and fibroblasts, while CP was primarily located in epithelial cells, and GSTO2 was minimally present in the epithelial cells and fibroblasts of nasal polyps. Consequently, a CeRNA regulatory network was constructed for the genes CP and GSTO2. This construction allowed for the prediction of potential drugs that could target CP. Conclusion:This study successfully identifies CP, SERPINF1 and GSTO2 as diagnostic and therapeutic markers related to oxidative stress in CRSwNP.

Objective:To explore the feasibility of constructing an objective tinnitus subtype model based on peripheral blood differentially expressed genes (DEGs) using a combination of Weighted Gene Co-expression Network Analysis (WGCNA) and Random Forest algorithm (RF).Methods:From October 2019 to June 2020, peripheral blood DEGs were obtained from 37 patients (from the Third Affiliated Hospital of Sun Yat-sen University)with chronic subjective high-frequency tinnitus (21 unbothersome type, 16 bothersome type) and 20 healthy volunteers through high-throughput sequencing. WGCNA was used to construct gene modules with different expression patterns and analyze their relationships with tinnitus characteristics. Subsequently, RF was employed to build subtype models, which were evaluated by the area under the receiver operating characteristic curve (AUC), accuracy, and F1-score.Results:A total of 12 351 intergroup DEGs were divided into 9 gene modules. Among them, MEblue, MEgreen, and MEbrown showed significant negative correlations with the healthy volunteer group, while MEpink showed a significant positive correlation with the tinnitus distress group. The "Tinnitus vs. Normal" and "Compensatory vs. Decompensatory" subtype models, based on MEblue and MEpink respectively, both had AUCs greater than 0.80, accuracies above 90%, and F1-scores above 0.90, indicating good performance.Conclusions:Peripheral blood DEGs are potential biological indicators for objective classification of subjective tinnitus. The combined application of WGCNA and the Random Forest algorithm should be a viable approach to constructing an objective tinnitus subtype model. However, further exploration and refinement are needed to validate the model′s generalizability, cross-dataset performance, and algorithm optimization.

Objective: To explore the association between greenness surrounding school and aggression among primary school students, and to explore the potential mediating roles of social support, loneliness, particulate matter (PM2.5) and Nitrogen Dioxide (NO2) in this association, in order to provide a scientific reference for preventing and ameliorating aggressive behaviors of primary school students. Methods: The data was used from a survey of children and adolescents conducted in 2015. The Chinese version of the Buss-Warren Aggression Questionnaire was used to assess total and subtypes of aggression, and the mean values of normalized difference vegetation index (NDVI) of 100 m, 500 m, 1 000 m circular buffers surrounding school were used to indicate the participants greenness exposure. PM2.5 and NO2 datas were obtained from the China High Air Pollutants Dataset.Generalized Linear Mixed Models were used to assess the associations of greenness surrounding school with total and subtypes of aggression. Results: Per IQR increment of NDVI-500 m [OR(95%CI)=1.09(1.03-1.15)] and NDVI-1 000 m[OR(95%CI)=1.07(1.02-1.13)] were positively correlated with physical aggression among primary school children, and per IQR increment of NDVI-100 m [OR(95%CI)=0.94(0.90-0.99)], NDVI-500 m [OR(95%CI)=0.93(0.89-0.98)] and NDVI-1 000 m [OR(95%CI)=0.95(0.91-1.00)] were negatively associated with verbal aggression (P<0.05). Mediation analyses revealed that social support partially mediated the association between the NDVI-500 m and physical aggression (mediation ratio:18.0%) and verbal aggression (mediation ratio:-8.3%) among primary school students, and loneliness partially mediated the association between the NDVI-500 m and physical aggression and verbal aggression among elementary school students effects, with proportion mediated ratios of -10.0% and 21.0%, respectively (P<0.05). Conclusions Exposure to school surrounding greenness is likely to associated with physical aggression and verbal aggression in primary school students, and social support and loneliness may partially mediate these associations.

AIM: To assess the accuracy of predicting intraocular lens(IOL)power after myopic refractive surgery using the Pentacam system's true net power(TNP)in the 3 mm zone combined with the SRK/T formula [i.e. TNP 3 mm(SRK/T)].METHODS: Retrospective study. This study enrolled 35 cases(50 eyes)of patients undergoing cataract surgery after laser assisted in situ keratomileusis(LASIK)or photorefractive keratectomy(PRK)from July 2019 to December 2021. Preoperatively, IOL power of 50 eyes, 34 eyes and 41 eyes was calculated by TNP 3 mm(SRK/T), Barrett True-K and Olsen 2 formulas, respectively, with at least 2 formulas used to calculate IOL power for each patient. The actual diopter was recorded 3 mo postoperatively. Prediction errors(PE)of IOL power were compared among the three calculation methods, and the proportion of eyes with PE within ±0.5 D and ±1.0 D was analyzed.RESULTS: The PE at 3 mo postoperatively for TNP 3 mm(SRK/T), Barrett True-K, and Olsen 2 was -0.02±0.63, -0.54±0.80, and 0.25±0.80 D, respectively(P<0.001). The proportions of PE within ±0.5 D were 66%(33/50), 44%(15/34)and 37%(15/41), respectively(P<0.05); the proportions of PE within ±1.0 D were 88%(44/50), 71%(24/34)and 80%(33/41), respectively(P>0.05).CONCLUSION: The Pentacam TNP 3 mm(SRK/T)method is simple to operate and provides accurate calculation of IOL power after corneal refractive surgery.