Objective To investigate how culture duration affects the expression of immune membrane proteins in human monocyte-derived dendritic cells (DCs) and their exosomes (DEXs). Methods Human monocytes were induced with recombinant granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin 4 (IL-4) to differentiate into DCs and were subsequently matured with tumor necrosis factor α(TNF-α). Exosomes were isolated by ultracentrifugation, and DEXs were identified by transmission electron microscopy and Amnis imaging flow cytometry, which were also used to quantify the expression of immune membrane proteins on DCs and DEXs. Results On the 10th day of culture, DCs displayed high surface expression of CD11c, CD80, CD86, major histocompatibility complex class I (MHC-I), and MHC-II. Expression peaked at day 18(CD11c: 78.66%±20.33%, CD80: 76.41%±10.02%, CD86: 96.43%±0.43%, MHC-I: 84.71%±2.96%, MHC-II: 80.01%±7.03%). After day 24, the overall expression showed a declining trend, with statistically significant differences observed for all markers except CD80 and MHC-II. By day 30, 80% of the DCs still expressed CD80, CD86, and MHC-II. The expression of immune membrane proteins on DEX surfaces also reached its peak on day 18, followed by an overall decline with prolonged culture time, with statistically significant differences observed for all markers except CD80. Correlation analysis revealed a significant positive relationship between the expression levels of immune membrane proteins on DC and DEX surfaces (CD11c: r=0.98; CD80: r=0.65; CD86: r=0.82; MHC-I: r=0.86; MHC-II: r=0.93). Conclusion Human monocyte-derived DCs in vitro express high expression of immune membrane proteins and maintain stable expression over a specific period. The exosomes secreted by these cells similarly demonstrate high surface expression of immune membrane proteins, with temporal trends aligned with those of the parent DCs.
Humans ; Dendritic Cells/immunology* ; Exosomes/immunology* ; Monocytes/metabolism* ; Cells, Cultured ; Time Factors ; B7-1 Antigen/metabolism* ; Membrane Proteins/immunology* ; Cell Culture Techniques/methods* ; B7-2 Antigen/metabolism* ; Cell Differentiation ; CD11c Antigen/metabolism* ; Granulocyte-Macrophage Colony-Stimulating Factor/pharmacology*

Objective This study aims to investigate the impact of cultivation time on dendritic cells(DCs)and their derived exosomes′ expression of immune-related membrane proteins(CD80,MHC-Ⅰ,MHC-Ⅱ)and provides experimental evidence for future research.Methods Mouse bone marrow cells were induced to differentiate into DCs using GM-CSF and IL-4,followed by maturation stimulation withTNF-α.Exosomes were extracted using ultracentrifugation.Western blot and Amnis image flow cytometry were used to identify exosomes derived from mouse DCs.Amnis image flow cytometry was used to detect the expression of immune-related proteins CD80,CD11c,MHC-Ⅰ,and MHC-Ⅱ in mouse DCs and their exosomes.Results After 5 days of in vitro cultivation,more than 50%of dendritic cells expressed CD80,CD11c,MHC-Ⅰ,and MHC-Ⅱ,reaching the highest level on day 13.The positivity rates were as follows:CD80(97.29±0.63)%,CD11c(92.31±1.18)%,MHC-Ⅰ(97.91±0.49)%,and MHC-Ⅱ(97.91±0.49)%.The differences were statistically significant(P<0.001).The expression gradually decreased after day 13,but approximately 80%of DC cells still expressed MHC-Ⅰ and MHC-Ⅱ immune molecules on day 30.The expression levels of CD80,CD11c,and MHC-Ⅱ on the exosome membrane were highest on day 5 and then decreased overall with prolonged cultivation time,except for MHC-Ⅰ molecules.The differences were statistically significant(P<0.01).Conclusions In vitro-cultured mouse dendritic cells express high levels of immune-related membrane proteins and can be stably maintained for a long time under suitable culture conditions.The secreted exosomes also carry abundant immune-related membrane proteins,but no significant correlation was found between the immune-related proteins on the dendritic cell surface and the exosome membrane surface.

Objective:To investigate the current application status of Cognitive Behavioral Therapy (CBT) techniques for Generalized Anxiety Disorder (GAD) by Chinese domestic experts using the Delphi method. The goal is to promote the standardization of CBT for GAD.Methods:Based on a review of relevant domestic and international literature, an expert consultation form including 59 CBT techniques for GAD (categorized into basic, cognitive, behavioral, and other techniques) was developed. The Delphi method was used to conduct two rounds of consultations with 24 qualified CBT experts. The application status and importance were ranked according to the statistical data.Results:(1) In the two rounds of consultation, the positivity index and the authority coefficient of the experts were both above 0.8. (2) After the second round of consultation, Kendall′s W coefficients of expert opinions on the frequency of use, contribution to treatment, and acceptance by clients were 0.397, 0.291, and 0.336, respectively, which are acceptable. Among the 59 CBT techniques for GAD, except for four behavioral techniques (music relaxation training,deliberately increasing anxiety experiments, systematic desensitization, and worry time), all other techniques received unanimous agreement from the experts in the three dimensions. (3) The top five techniques in terms of comprehensive application were data collection, establishing a therapeutic alliance, clinical psychological assessment, psychoeducation, and goal setting. Conclusion:In CBT for GAD, domestic experts unanimously recognize the importance and applicability of basic, cognitive, and traditional behavioral CBT techniques. However, they are relatively cautious about the clinical application of emerging techniques from the third generation of CBT, except for mindfulness and sleep management.

Objective:To investigate the current application status of Cognitive Behavioral Therapy (CBT) techniques for Generalized Anxiety Disorder (GAD) by Chinese domestic experts using the Delphi method. The goal is to promote the standardization of CBT for GAD.Methods:Based on a review of relevant domestic and international literature, an expert consultation form including 59 CBT techniques for GAD (categorized into basic, cognitive, behavioral, and other techniques) was developed. The Delphi method was used to conduct two rounds of consultations with 24 qualified CBT experts. The application status and importance were ranked according to the statistical data.Results:(1) In the two rounds of consultation, the positivity index and the authority coefficient of the experts were both above 0.8. (2) After the second round of consultation, Kendall′s W coefficients of expert opinions on the frequency of use, contribution to treatment, and acceptance by clients were 0.397, 0.291, and 0.336, respectively, which are acceptable. Among the 59 CBT techniques for GAD, except for four behavioral techniques (music relaxation training,deliberately increasing anxiety experiments, systematic desensitization, and worry time), all other techniques received unanimous agreement from the experts in the three dimensions. (3) The top five techniques in terms of comprehensive application were data collection, establishing a therapeutic alliance, clinical psychological assessment, psychoeducation, and goal setting. Conclusion:In CBT for GAD, domestic experts unanimously recognize the importance and applicability of basic, cognitive, and traditional behavioral CBT techniques. However, they are relatively cautious about the clinical application of emerging techniques from the third generation of CBT, except for mindfulness and sleep management.

Objective:To analyze the distribution characteristics of plasma renin concentration (PRC) in patients with aldosterone-producing adenoma (APA) and its impact on diagnosis.Methods:In this retrospective case series, clinical data from 200 patients with APA (80 men and 120 women; mean age 45.6 years) in the First Affiliated Hospital of Chongqing Medical University from November 2013 to January 2022 were evaluated. PRC was determined by automated chemiluminescence immunoassay. The distribution characteristics of PRC were analyzed, and 8.2 mU/L was used as the low renin cutoff to evaluate whether renin was suppressed.Results:The median PRC was 1.6 mU/L (range, 0.4-41.5 mU/L). There were 116 patients with APA with PRC of ≤2 mU/L, 41 patients with 28.2 mU/L) in 8.0% (16/200) of the patients with APA. And PRC was not suppressed in 2.5% (5/200) of the patients with APA, resulting in a primary aldosteronism negative screening outcome.Conclusions:Although most patients with APA have low PRC, there are a small number (8%) of patients whose PRC has not been fully suppressed, which can lead to missed diagnoses during primary aldosteronism screening. While primary aldosteronism is highly suspected, further investigations are required to determine the diagnosis, even if PRC is not fully suppressed at screening.

Objective:To investigate the expression of ras-related C3 botulinum toxin substrate 3 (RAC3) in glioma tissues and its effect on the migration and invasion of glioma cells.Methods:The expression of RAC3 in 57 glioma patients and their adjacent tissues from the First People’s Hospital of Shangqiu was detected by immunohistochemical assay. According to the experimental requirements, brain glioma cells U87MG were divided into experimental group and control group. The experimental group U87MG cells were transfected with RAC3-siRNA plasmid, and the control group U87MG cells were transfected with MOCK-siRNA plasmid. RAC3 mRNA in each group was detected by fluorescence quantitative PCR. The expressions of RAC3 and MMP2 in each group were detected by Western blot. Transwell was used to detect the migration and invasion ability of cells in each group.Results:The positive rate of RAC3 in glioma patients was 89.47% (51/57 cases) , and the expression rate in paracancer tissues was 14.04% (8/57 cases) . The expression rate of RAC3 in glioma tissues was significantly higher than that in paracancer tissues, with statistical significance ( P<0.01) . After siRNA transfection, mRNA expression of RAC3 in experimental group and control group was 1.23±0.20 and 0.43±0.12, and protein expression of RAC3 was 1.19±0.11 and 0.23±0.08, respectively. The expression of MMP2 protein was 1.19±0.11 and 0.23±0.08, respectively. The expression of MMP2 in experimental group was significantly decreased ( P<0.05) . Transwell assay showed that the number of invasive cells in experimental group and control group U87MG cells was (22±5) and (45±8) , and the number of migratory cells was (34±6) and (90±11) , respectively. In experimental group, U87MG cell migration and invasion ability decreased significantly (both P<0.05) . Conclusion:The high expression of RAC3 in glioma tissues may be related to the malignant degree of development, and affect the migration and invasion ability of glioma cells by regulating the expression of MMP2.

Deficiency, stasis, water and toxin are of great significance in the pathogenesis and pathologic evolution of chronic heart failure (CHF). Based on "deficiency, blood stasis, water and toxin", the pathogenesis and treatment of CHF were discussed in this article. It was found that in the pathogenesis, deficiency--deficiency of heart qi and deficiency of heart yang were the origin of the disease, and blood stasis, water and toxin were the markers of the disease. Among them, blood stasis was the central pathological link, and also an important mechanism that could aggravate the disease and cause a vicious cycle; water-phlegm and water dampness were the basic pathological products; toxin-heat toxin, water toxin, and stasis toxin were the final results of disease progress and product accumulation. In terms of treatment, CHF can be divided into four stages: early, middle, late and end. In the early stage, tonifying qi and regulating heart can be used for the treatment of root cause, and promoting blood circulation and water can be used for the treatment of symptoms; tonifying qi and yin and reinforcing the healthy qi, reducing blood stasis, purging turbid, and eliminating pathogenic factors can be used in the middle stage; reducing blood stasis and removing toxic materials should be used in the late stage, supplemented with warming yang and increasing urine excretion; astringing yang,generating body fluids, tonifying qi and yang should be used in the end stage. At the same time of treating by stages, attention should be paid to adhering to a holistic concept and dialectical treatment; pay attention to timing and flexible medication; adopting a combination of Chinese and Western approaches and integrating them.

Pulmonary blast injury has become the main type of trauma in modern warfare, characterized by externally mild injuries but internally severe injuries, rapid disease progression, and a high rate of early death. The injury is complicated in clinical practice, often with multiple and compound injuries. Currently, there is a lack of effective protective materials, accurate injury detection instrument and portable monitoring and transportation equipment, standardized clinical treatment guidelines in various medical centers, and evidence-based guidelines at home and abroad, resulting in a high mortality in clinlcal practice. Therefore, the Trauma Branch of Chinese Medical Association and the Editorial Committee of Chinese Journal of Trauma organized military and civilian experts in related fields such as thoracic surgery and traumatic surgery to jointly develop the Clinical treatment guideline for pulmonary blast injury ( version 2023) by combining evidence for effectiveness and clinical first-line treatment experience. This guideline provided 16 recommended opinions surrounding definition, characteristics, pre-hospital diagnosis and treatment, and in-hospital treatment of pulmonary blast injury, hoping to provide a basis for the clinical treatment in hospitals at different levels.

Objective:The aim of the present study was to re-evaluate the diagnostic value and optimal cutoff point of captopril challenge test (CCT) in diagnosis of primary aldosteronism (PA).Methods:This is a retrospective study. All patients with a high risk for PA underwent screening test, and then proceeded to CCT and fludrocortisone suppression test (FST) on different days. The FST was used as a reference standard for PA. The plasma renin concentration (PRC) and plasma aldosterone concentration (PAC) were measured with an automated chemiluminescence immunoassay. Random number method was performed in the patients with unilateral primary aldosteronism (UPA), in order to make the proportion of the analyzed UPA in PA was 35%. Receiver operating characteristic (ROC) analyses were performed to compare diagnostic accuracy.Results:A total of 543 patients with 400 PA patients and 143 essential hypertension (EH) patients were enrolled. The diagnostic value of post-CCT PAC was significantly higher than that of the post-CCT plasma aldosterone-renin ratio (ARR), and that of the PAC suppression percentage, respectively. The area under the ROC curve (AUC ROC) was 0.86 (0.83, 0.89) for PAC, 0.78 (0.74, 0.82) for ARR, and 0.62 (0.56, 0.67) for the PAC suppression percentage (all P<0.01), respectively. The optimal cutoff point of post-CCT PAC for PA was 110 ng/L, in which the sensitivity and specificity were 73.25% and 79.02%, respectively. The diagnostic efficiency of post-CCT PAC was not improved either in combination with PAC suppression percentage or in combination with post-CCT ARR. Conclusions:CCT is a useful test for the confirmation of PA. PAC level of 110 ng/L at 2 h after 50 mg of captopril is recommended as an optimal cutoff point for the diagnosis of PA.

Objective:To investigate the changes in the level of microRNA-193b-3p（miR-193b-3p）in patients with aneurysmal subarachnoid hemorrhage（aSAH）after hyperbaric oxygen（HBO）treatment and to explore its significance.Methods:A total of 176 patients with aSAH admitted to the Emergency Department of the First People’s Hospital of Shangqiu from January 2019 to September 2020 were selected as the research subjects. They were divided into sound prognosis group（ n=128）and poor prognosis group（ n=48）according to scores of the modified Rankin Scale（mRS）90 days after treatment. The relative expression levels of miR-193b-3p were detected by real-time quantitative PCR（RT-qPCR）. The prognostic risk factors of aSAH patients were analyzed by logistic regression. The value of miR-193b-3p in judging the prognosis of aSAH patients was evaluated by receiver operating characteristic（ROC）curve. The dose-response relationship between miR-193b-3p and prognosis in aSAH patients was analyzed by the logistic regression using restricted cubic splines. Results:The relative expression levels of miR-193b-3p in the sound prognosis group before and after treatment were higher than those in the poor prognosis group at the same time points（ P<0.05），and the relative expression levels of miR-193b-3p in both groups were decreased after treatment（ P<0.05）. The area under the ROC curve（AUC），sensitivity，and specificity of post-treatment miR-193b-3p in judging the prognosis of aSAH patients were 0.818，68.75%，and 85.94%，respectively. Hunt-Hess Ⅲ-Ⅳ，cerebral vasospasm（CVS），Glasgow Coma scale（GCS）rating，and high C-reactive protein（CRP）level were independent risk factors for aSAH prognosis（ P<0.05）；the relatively high expression level of post-treatment miR-193b-3p was an independent protective factor for aSAH prognosis（ P<0.05）. Post-treatment miR-193b-3p had a non-linear relationship with the prognosis of aSAH（ P<0.05）. Conclusion:After HBO treatment，the relative expression level of miR-193b-3p in aSAH patients was decreased，which is related to the poor prognosis of aSAH.