Objective To explore macrophage subpopulations in ischemic stroke (IS) by using single-cell RNA sequencing (scRNA-seq) data analysis and High-Dimensional Weighted Gene Co-Expression Network Analysis (hdWGCNA). Methods Based on single-cell sequencing data, transcriptomic information for different cell types was obtained, and macrophages were selected for subpopulation identification. hdWGCNA, cell-cell communication, and pseudotime trajectory analysis were used to explore the characteristics of macrophage subpopulations following IS. Key genes related to IS were identified using microarray data and validated for diagnostic potential through Receiver Operating Characteristic (ROC) analysis. Gene Set Enrichment Analysis (GSEA) was conducted to investigate the potential functions of these genes. Results The scRNA-seq data analysis revealed significant changes in macrophage subpopulation composition after IS. A specific macrophage subpopulation enriched in the stroke group was identified and designated as MCAO-specific macrophages (MSM). Pseudotime trajectory analysis indicated that MSM cells were in an intermediate stage of macrophage differentiation. Cell-cell communication analysis uncovered complex interactions between MSM cells and other cells, with the CCL6-CCR1 signaling axis potentially playing a crucial role in neuroinflammation. Two gene modules associated with MSM were identified via hdWGCNA, significantly enriched in pathways related to NOD-like receptors and antigen processing. By integrating differentially expressed MSM genes with conventional transcriptomic data, three IS-related hub genes were identified: Arg1, CLEC4D, and CLEC4E. Conclusion This study reveals the characteristics and functions of macrophage subpopulations following IS and identifies three hub genes with potential diagnostic value, providing novel insights into the pathological mechanisms of IS.
Macrophages/metabolism* ; Computational Biology/methods* ; Single-Cell Analysis/methods* ; Transcriptome ; Ischemic Stroke/metabolism* ; Animals ; Gene Regulatory Networks ; Gene Expression Profiling ; Humans ; Male

Objective:To investigate the value of neutrophil lymphocyte ratio (NLR) and systemic immune inflammatory index (SII) for the diagnosis of coronary artery lesions in patients with acute phase of Kawasaki disease (KD).Methods:It was a cross-sectional study. Children with acute Kawasaki disease admitted to Linyi Central Hospital from January 2018 to December 2021 were enrolled and divided into the Kawasaki disease complicated with coronary artery disease group (KD-CAL group) and the Kawasaki disease without coronary artery disease group (KD-NCAL group). The demographic data, blood routine and C-reactive protein (CRP) test results before intravenous injection of γ globulin were collected, and NLR and SII were calculated. Multivariate logistic regression model was used to analyze the risk factors of coronary artery disease in children with Kawasaki disease. Receiver operating characteristic (ROC) curve was used to analyze the diagnostic value of NLR and SII for coronary artery lesions in children with Kawasaki disease.Results:A total of 109 children with acute phase of KD aged 1 month to 9 years were enrolled, including 66 boys (60.6%). There were no significant differences in age and gender composition between the KD-CAL group ( n=17) and the KD-NCAL group ( n=92) (all P>0.05). Compared with the KD-NCAL group, the NLR, SII and CRP levels in the KD-CAL group were higher (all P<0.05). Multivariate logistic regression analysis showed that NLR and SII were independent risk factors of coronary artery disease in children with Kawasaki disease (NLR, OR=1.265, 95% CI:1.066-1.502, P=0.007; SII, OR=1.001, 95% CI:1.000-1.002, P=0.015). ROC curve results showed that thearea under the curve ( AUC) of NLR in diagnosis of coronary artery lesions was 0.812 (95% CI:0.703-0.921, P<0.05), taking 5.4 as cutoff value, the sensitivity was 73.9% and specificity was 76.5%; the AUC of SII in diagnosis of coronary artery lesions was 0.830 (95% CI:0.741-0.919, P<0.05), taking 1 623 as the cut-off value, the sensitivity was 66.3% and specificity was 88.2%; the AUC of the combination of NLR and SII was 0.875(95% CI:0.790-0.959, P<0.05), the sensitivity and specificity were 85.9% and 76.5%, respectively. Conclusion:NLR and SII have certain diagnostic value for coronary artery disease in children with acute Kawasaki disease, and the combination of NLR and SII has better value than single index.

Objective To investigate the relationship between interleukin-1β (IL-1β) and miR-185-5p in the process of joint injury in acute gouty arthritis (AGA). Methods The serum miR-185-5p levels of 89 AGA patients and 91 healthy volunteers were detected by real-time quantitative PCR. The correlation between miR-185-5p expression level and VAS score or IL-1β expression level was evaluated by Pearson correlation coefficient method. Receiver operating characteristic (ROC) curve was used to evaluate the diagnostic value of miR-185-5p in AGA. THP-1 cells were induced by sodium urate (MSU) to construct an in vitro acute gouty inflammatory cell model. After the expression level of miR-185-5p in THP-1 cells was upregulated or downregulated by transfection of miR-185-5p mimics or inhibitors in vitro, inflammatory cytokines of THP-1 cells, such as IL-1β, IL-8 and tumor necrosis factor α (TNF-α), were detected by ELISA. The luciferase reporter gene assay was used to determine the interaction between miR-185-5p and the 3'-UTR of IL-1β. Results Compared with the healthy control group, the expression level of serum miR-185-5p in AGA patients was significantly reduced. The level of serum miR-185-5p was negatively correlated with VAS score and IL-1β expression level. The area under the curve (AUC) was 0.905, the sensitivity was 80.17% and the specificity was 83.52%. Down-regulation of miR-185-5p significantly promoted the expression of IL-1β, IL-8 and tumor necrosis factor (TNF-α), while overexpression of miR-185-5p showed the opposite results. Luciferase reporter gene assay showed that IL-1β was the target gene of miR-185-5p, and miR-185-5p negatively regulated the expression of IL-1β. Conclusion miR-185-5p alleviates the inflammatory response in AGA by inhibiting IL-1β.
Humans ; 3' Untranslated Regions ; Arthritis, Gouty/genetics* ; Interleukin-1beta/genetics* ; Interleukin-8 ; Luciferases ; MicroRNAs/genetics* ; Tumor Necrosis Factor-alpha

AIM To optimize the extraction process for Jigen Standard Decoction,and to establish its quality standard.METHODS With soaking time,water addition and first decoction time as influencing factors,comprehensive score for 3,6'-disinapoyl sucrose content and yield rate as an evaluation index,the extraction process was optimized by response surface method on the basis of single factor test.The content and transfer rate of 3,6'-dimustayl sucrose were determined,after which HPLC characteristic chromatograms were established,cluster analysis,principal component analysis and orthogonal partial least squares discriminant analysis were performed.RESULTS The optimal conditions were determined to be 60 min for soaking time,(12+11)times for water addition,and(47+20)min for decoction time,the comprehensive score was 97.98.Fifteen batches of standard decoctions demonstrated the average yield rate and transfer rate of 14.182％and 20.468％,respectively,whose characteristic chromatograms existed six common peaks with the similarities of more than 0.9(except for S4,S8).Various batches of standard decoctions were clustered into two types,three principal components displayed the acumulative variance contribution rate of 91.4％,peaks 2,6 were quality markers.CONCLUSION This precise,stable and reproducible method can be used for the preparation and quality control of Jigen Standard Decoction.

OBJECTIVE: To investigate the clinical characteristics and treatment of pneumocystis carinii pneumonia (PCP) in children with acute lymphoblastic leukemia (ALL), in order to improve the early diagnosis and effective treatment. METHODS: Clinical data of five children with ALL developing PCP in the post-chemotherapy granulocyte deficiency phase were analyzed retrospectively. The clinical manifestations, laboratory tests, imaging findings, treatment methods and effect were summarized. RESULTS: The male-to-female ratio of the five children was 1∶4, and the median age was 5.5 (2.9-8) years old. All patients developed PCP during granulocyte deficiency phase after induction remission chemotherapy. The clinical manifestations were generally non-specific, including high fever, tachypnea, dyspnea, non-severe cough, and rare rales in two lungs (wet rales in two patients). Laboratory tests showed elevated C-reactive protein (CRP), serum procalcitonin (PCT), (1,3)-β-D-glucan (BDG), lactate dehydrogenase (LDH) and inflammatory factors including IL-2R, IL-6 and IL-8. Chest CT showed diffuse bilateral infiltrates with patchy hyperdense shadows. Pneumocystis carinii(PC) was detected in bronchoalveolar lavage fluid (BALF) or induced sputum by high-throughput sequencing in all patients. When PCP was suspected, chemotherapy was discontinued immediately, treatment of trimethoprim-sulfame thoxazole (TMP-SMX) combined with caspofungin against PC was started, and adjunctive methylprednisolone was used. Meanwhile, granulocyte-stimulating factor and gammaglobulin were given as the supportive treatment. All patients were transferred to PICU receiving mechanical ventilation due to respiratory distress during treatment. Four children were cured and one died. CONCLUSION PCP should be highly suspected in ALL children with high fever, dyspnea, increased LDH and BDG, and diffuse patchy hyperdense shadow or solid changes in lung CT. The pathogen detection of respiratory specimens should be improved as soon as possible. TMP/SMZ is the first-line drug against PCP, and the combination of Caspofungin and TMP/SMZ treatment for NH-PCP may have a better efficacy. Patients with moderate and severe NH-PCP may benefit from glucocorticoid.
Caspofungin/therapeutic use* ; Child ; Child, Preschool ; Dyspnea ; Female ; Humans ; Male ; Pneumonia, Pneumocystis/therapy* ; Precursor Cell Lymphoblastic Leukemia-Lymphoma/therapy* ; Respiratory Sounds ; Retrospective Studies

OBJECTIVE: To retrospectively analyze the identification results of irregular antibodies, to clarify the distribution features and to explore the relation of alloantibodies and autoantibodies with the immunized history of patients and disease kinds. METHODS: 49 820 patients who applied for red blood transfusion during Sep 1st 2017 to Sep 1st 2018 were selected. All the specimens were screened for the antibody by microcolumn gel antiglobulin technique, which then were identified for irregular antibody. RESULTS: Antibodies were found in 861 (1.73%) of all 49 820 transfused samples. The alloimmunization history of the patients with antibodies was significantly different between male and female (χ=18.54，P＜0.01). The alloantibody was the most common, accounting for 59.50% in all of the antibodies. Warm autoantibody, anti-E, anti-M, anti-cE and anti-Ce accounted for 68.5% of the antibodies. The blood group of Rh, MNS and Lewis were responsible for 92.40% of alloantibody, especially anti-E accounted for the largest percentage(38.60%) of alloantibody. Patients with alloantiboies experienced much more the alloimmunization and transfusion history (χ=20.13，P＜0.01；χ=5.40，P＜0.05) . The distribution of auto and alloantibody was very significantly different among the ddifferent isease (χ=51.8，P＜0.01), Hematopathy, solid tumor and osteoarthropathy were often associated with alloantibody, otherwise, autoantibodies often occurred in hematopathy and autoimmune disease. CONCLUSION The most important factor that results in antibody-screening positive is alloantibody, in which anti-E antibody from Rh blood group system in most common.
Antibodies ; immunology ; Blood Group Antigens ; Blood Transfusion ; Erythrocytes ; Female ; Humans ; Isoantibodies ; Male ; Retrospective Studies

Objective    To compare the efficacy of percutaneous closure guided by transthoracic echocardiography or angiography in the treatment of patent ductus arteriosus (PDA). Methods    Literature databases such as CNKI, VIP, Wanfang Database, PubMed, Cochrane Library were searched for collecting published literatures on percutaneous closure for PDA guided by transthoracic echocardiography and angiography, retrieval time limit was up to April 2019. Two evaluators independently screened the literature, extracted the data and evaluated the quality according to inclusion and exclusion criteria. The collected data were analyzed by RevMan 5.3 software. Results    Eight studies were included finally, with a total sample size of 681 cases. Meta-analysis showed that there was no statistical difference in the operative success rate between the echocardiography group and the angiography group (RR=0.99, 95%CI 0.97- 1.01, P=0.40). Postoperative complications were less in the echocardiography group than those in the angiography group (RR=0.26, 95%CI 0.11-0.59, P=0.001).The operation time (P<0.000 01), amount of intraoperative radiation (P< 0.000 01), exposure time (P<0.000 01), hospitalization days (P<0.000 01) and hospitalization costs (P<0.000 01) in the echocardiography group were less or shorter than those in the angiography group, and the difference was statistically different. Conclusion    Compared with angiography-guided, transthoracic echocardiography-guided percutaneous closure for PDA is a safe and effective method with less trauma, lower cost, and can replace angiography as one of the guiding methods for PDA.

OBJECTIVETo assess the value of Prostate Imaging and Reporting and Data System: Version 2 (PI-RADS v2) combined with prostate specific antigen (PSA) in the diagnosis of peripheral zone (PZ) prostate cancer (PCa).

METHODSThe preoperative magnetic resonance imaging and PSA data were ananlyzed for 69 patients with pathologically confirmed PCa and 109 non-PCa patients. PI-RADS v2 scores (1-5) was used to evaluate the risk of PZ PCa. The total PSA (tPSA) level, free to total PSA ratio (f/t PSA), PSA density (PSAD), PZ-PSAD and PI-RADS v2 scores were compared between the PCa and non-PCa patients. Logistic regression models were established with parameters that differed significantly the two groups. The receiver opearting characteristics (ROC) curve was constructed based on the P values derived from the logical regression models and PI-RADS scores to assess the diagnostic efficiency.

RESULTSPI-RADS v2 score, tPSA, f/t PSA, PSAD and PZ-PSAD differed significantly between the two groups (P<0.01). Four predictive multivariate models were established: Logit P=-6.825+1.024PI-RADS v2+ 0.223tPSA (A), Logit P=-4.354+1.586PI-RADS v2-12.7841f/tPSA (B), Logit P=-8.993+1.630PI-RADS v2+17.091PSAD (C), and Logit P=-9.434+1.596PI-RADS v2+10.494PZ-PSAD (D), whose area under the ROC curves was 0.908, 0.891, 0.944, and 0.961, respectively, all significantly greater than that of PI-RADS v2 score (P<0.05).

CONCLUSIONCompared with PI-RADS v2 score alone, the combination of PI-RADS v2 score and PSA in the logistic regression model can improve the diagnostic efficiency of PZ PCa and offers better confidence in the decision of biopsy in suspected cases.

OBJECTIVETo evaluate the clinical value of modified Bethesda assay, modified Nijmegen assay and blank assay for detection of coagulation factor VIII (FVIII) inhibitors in patients with hemophilia A and analyze the factors that affect FVIII inhibitor detection.

METHODSThe levels of FVIII inhibitors in 257 patients with hemophilia A were detected using modified Bethesda assay, modified Nijmegen assay and blank assay (in which the buffer or FVIII-deficient plasma in the control mixture was replaced by deionized water). The 3 methods were compared for positivity rates and FVIII inhibitor titers based on the positive cut-off value of FVIII inhibitors ≥0.60 BU/mL.

RESULTSThe positive rates of modified Bethesda assay, modified Nijmegen assay and blank assay were 79.38%, 85.21% and 72.37%, respectively. A strong correlation was found between the results by modified Bethesda assay and modified Nijmegen assay (r=0.996, P<0.001), and FVIII inhibitor titers (P<0.001) but not the positive rates (P=0.105) detected by the two methods differed significantly. The correlation coefficients between modified Nijmegen assay and blank assay was 0.994 (P<0.001), and a significant difference was found in FVIII inhibitor titers (P<0.001) but not the positivity rates (P=0.079) detected by the two methods. The correlation coefficient between modified Nijmegen assay and blank assay was 0.994 (r=0.994), and the two methods yielded significantly different FVIII inhibitor titers and positivity rates (P=0.001).

CONCLUSIONThe modified Bethesda assay has a lower sensitivity than modified Nijmegen assay but has a higher sensitivity than blank assay. The consistency level of coagulation factors in the reaction system and stable buffer system are important factors that affect FVIII-inhibitor detection.

Objective To investigate the job satisfaction of nurses from one level three class A stomatology hospital and explore its influencing factors. Methods A questionnaire-based survey was used to examine job satisfaction of 791 nurses from the stomatology hospital in Beijing and analyze its influencing factors. Results A total of 656 effective questionnaire were collected with effective collective rate 82. 9%. The total average score of job satisfaction was (3. 24 ± 0. 92), which was at or above intermediate-level; dissatisfaction dimensions contained extrinsic rewards, family/work balance and professional opportunities. The correlation analysis showed job satisfaction was negatively correlated with age, working years and years serving in this hospital (P<0. 01). Single-factor analysis showed nurses working in different department, with different levels of formal educations, professional titles and turnover intention varied significantly in job satisfaction (P<0. 05); multiple regression analysis showed that turnover intention and age might be influencing factors of job satisfaction. Conclusions The overall job satisfaction of dental nurses is at upper level. Efficient way to increase job satisfaction may be reducing turnover intention, improving extrinsic rewards and professional opportunities.