Cryopreservation is the best preserving method of the homograft heart valve at present, but at the same time it may damage the valve structural integrity. This study was performed to probe into the effects of cryopreservation on the histology of aortic valve. The viability of the cardiac valves was evaluated by XTT-tetrazolium colorimetric assay and the histologic property was assayed by light microscopy, transmission electron microscopy, and indirect immunofluorescent examination. The results of the experiments showed that the valve cellular viability was markedly decreased (P < 0.05), the cellular ultrastructure was damaged, the soluble extracellular matrix fibronectin and chondroitin sulfate were leached at different degree, and the interstitial collagen fibers were irregularly aligned after cryopreservation, but the damaged degree has no relation to storage within a period of 3 months.
Aortic Valve ; Cryopreservation ; Extracellular Matrix ; ultrastructure ; Heart Valve Prosthesis ; Humans ; Transplantation, Homologous

To investigate the feasibility of making up bovine pericardium into scaffolds for tissue engineering valve, it was decellularized by 0.5% trypsin incubation for three hours. The rats were divided into fresh bovine group, acellular pericardium group and control group, and OD value of rat anti-bovine IgG was measured. Serum levels of IgG showed no difference among the three groups on the 7th day postoperatively (P > 0.05). The OD values of IgG at other time-points displayed distinct difference among the three groups (P < 0.05). Fresh bovine pericardium group produced more IgG than acellular bovine pericardium group did at the same time-point. The IgG level of fresh bovine pericardium group reached the peak postoperatively, but in acellular bovine pericardium group its peak value appeared on the 30th day postoperatively. Histological examination demonstrated that the bovine pericardium was thoroughly decellularized, the fibrous structure was slightly loose, but remained essentially intact. These results indicate that the acellularization procedure can result in a complete removal of cells and decrease of immunogenicity.
Animals ; Antibodies, Anti-Idiotypic ; immunology ; Bioprosthesis ; Cattle ; Cell Separation ; methods ; Immunoglobulin G ; immunology ; Implants, Experimental ; Pericardium ; cytology ; immunology ; Rats ; Rats, Sprague-Dawley ; Tissue Engineering ; methods ; Trypsin ; pharmacology

Objective:This study aims to establish an early warning diagnosis model for acute exacerbation of chronic obstructive pulmonary disease (AECOPD) and to provide a simple, rapid, and accurate auxiliary diagnosis basis for clinical practice.Methods:The sample bank of subjects (patients admitted to the Eighth Medical Center of the PLA General Hospital from September 10, 2021, to July 25, 2023) was constructed, including the model establishment cohort [SCOPD group 49, 42 males and 7 females, (69.71±11.16) years old; AECOPD group 53, 49 males and 4 females, (72.60±10.19) years old] and the model validation cohort [SCOPD group 35, 28 males and 7 females, (69.97±10.40) years old; AECOPD group 35, 33 males and 2 females, (71.43±9.67) years old]. Fasting peripheral blood samples were collected, and the expression levels of IL-5, IL-17A, and IFN-α were detected by flow cytometry. Different expression levels were analyzed by Mann-Whitney U test. Binary logistic regression analysis was used to screen the related risk factors of COPD patients in acute exacerbation. The diagnostic efficacy of the model was evaluated by the receiver operating characteristic (ROC) curve.Results:The levels of IL-5 [1.64 (0.60, 2.86) pg/ml], IL-17A [1.42 (0.88, 2.29) pg/ml], and IFN-α [0.91 (0.59, 1.81) pg/ml] in the SCOPD group were significantly decreased compared with the AECOPD group IL-5 [4.68 (2.34, 9.40) pg/ml, Z=-5.033, P<0.001], IL-17A [2.33 (1.59, 4.62) pg/ml, Z=-3.919, P<0.001], IFN-α [2.83 (0.91, 3.75) pg/ml, Z=-4.127, P<0.01] in the cohort of model establishment. The results of binary logistic regression analysis between SCOPD and AECOPD groups showed that IL-5, IL-17A, and IFN-α were independent risk factors for acute exacerbation of patients with COPD ( P<0.05). And the regression equation is Y=-2.861+0.364×IL-5+0.385×IL-17A+0.445×IFN-α. The AUC value of IL-5, IL-17A, IFN-α and combined detection was 0.866 ( P<0.001). Compared to the SCOPD group and the AECOPD group in the cohort of model validation, the receiver operating characteristic (ROC) curve showed that the combined model of three (AUC=0.858, P<0.001) could be used to diagnose the AECOPD. And the Kappa value was 0.773( P<0.05). Conclusion:The combined detection of IL-5, IL-17A, and IFN-α has high diagnostic efficacy for patients with acute exacerbation of COPD. This method provides a new potential tool for the clinical diagnosis of AECOPD and has the value of further exploration and optimization, promotion, and application.

Objective:To analyze the results of lymphocyte subsets and 12 plasma cytokines in patients with tuberculosis by flow cytometry and to evaluate their diagnostic efficacy in these patients.Methods:This is a retrospective case-control study. A total of 128 patients with evidence of tuberculosis disease or clinically confirmed tuberculosis who were admitted to the 8th Medical Center of PLA General Hospital from January 2022 to December 2023 were included. According to the location of mycobacterium tuberculosis infection, the patients were divided into the pulmonary tuberculosis group (83 cases) and the extrapulmonary tuberculosis group (45 cases), and 100 healthy age-and sex matched people who underwent health check up during the study period were selected as the control group. Flow cytometry was used to detect peripheral blood lymphocyte subsets and 12 plasma cytokines [including 10 pro-inflammatory factors: interleukin (IL)-5, interferon (IFN)-α, IL-2, IL-6, IL-1β, IFN-γ, IL-8, IL-17, IL-12P70, Tumor necrosis factor (TNF)-α, and two anti-inflammatory factors: IL-4, IL-10] in participants of all groups. Spearman correlation method was used to analyze the correlation between lymphocyte subsets and cytokines, binary Logistic regression was used to screen the TB related factors, and receiver operating curve (ROC) was used to evaluate the diagnostic efficacy of TB related factors.Results:Compared with the control group, the absolute number of CD3 +T lymphocytes, CD3 +CD8 +T lymphocytes, CD3 +CD4 +T lymphocytes, NK cells and B cells were lower in pulmonary tuberculosis group and extrapulmonary tuberculosis group (all P<0.05). Except for IL-1β, the levels of other 11 cytokines are all significantly higher in the pulmonary tuberculosis group (all P<0.01), and the levels of IL-6, IFN-γ, IL-17, TNF-α, IL-4 and IL-10 were significantly higher in extrapulmonary tuberculosis group (all P<0.05). Compared with extrapulmonary tuberculosis group, the level of IL-8 was higher in pulmonary tuberculosis group ( P=0.026). Spearman correlation analysis showed that IL-6, IFN-γ and IL-8 were negatively correlated with the absolute numbers of CD3 +T lymphocytes, CD3 +CD8 +T lymphocytes, CD3 +CD4 +T lymphocytes, NK cells and B cells (IL-6: R2=-0.30, -0.28, -0.32, -0.26, -0.28; IFN-γ: R2=-0.36, -0.31, -0.37, -0.25, -0.36; IL-8: R2=-0.14, -0.13, -0.16, -0.14, -0.22; all P<0.05), IL-10 was negatively correlated with the absolute number of CD3 +CD4 +T lymphocytes, NK cells and B cells ( R 2=-0.14, -0.19, -0.21, all P<0.05); Binary Logistic regression analysis showed that IL-6, IFN-γ, IL-8 and IL-10 were the related factors of tuberculosis ( OR=1.809, 1.136, 0.910, 2.218, all P<0.05), ROC curve analysis showed that the AUC of IL-6, IFN-γ, IL-8 and IL-10 in the joint diagnosis of tuberculosis was 0.845, the sensitivity was 0.766, and the specificity was 0.820. Conclusion:The lower absolute number of lymphocyte subsets and cytokine levels in patients with pulmonary tuberculosis and extrapulmonary tuberculosis indicate that their immune function is in a low state, and the higher levels of pro-inflammatory factors (IL-6, IFN-γ, IL-8) and anti-inflammatory factor (IL-10) indicates the higher inflammatory status, and evaluation of these 4 cytokines has satisfactory diagnostic efficacy for tuberculosis.