【摘要】 目的 比较密度梯度离心法及全骨髓培养法分离培养内皮祖细胞的差异。方法 取4周雄性近交系C57BL/6J小鼠骨髓,分别使用密度梯度离心法及全骨髓培养法培养,观察细胞贴壁情况和细胞形态,并行DiIacLDL及FITCUEAI双染、vWF、eNOS及细胞表面标志检测。结果 密度梯度离心法培养细胞可形成典型铺路石样改变及形成血管样结构;而全骨髓培养法贴壁细胞形态多样,较多呈长梭形铺展生长,部分细胞呈类圆形及纺锤形。比较两种方法培养细胞摄取DiIacLDL、结合FITCUEAI双阳性率以及vWF、eNOS及细胞表面标志表达阳性率,差别均有统计学意义(Plt;005)。应用密度梯度离心法,随着培养时间延长,表达CD34、CD133及FLk1细胞逐渐增多(Plt;005)。结论 密度梯度离心法及全骨髓培养法在EGM2MV培养体系下均可培养出内皮祖细胞,但密度梯度离心法较全骨髓培养法培养的内皮祖细胞纯度高。
目的:研究CC亚族趋化因子单核细胞趋化蛋白-4(MCP-4/CCL13)在类风湿关节炎(RA)患者外周血的表达水平,并分析MCP-4的水平与疾病活动性的关系,以探讨MCP-4在RA发病机制中的作用。 方法:应用酶联免疫吸附实验(ELISA)定量方法测定40例RA患者、20例其它自身免疫性疾病患者和20例正常健康对照者血清MCP-4水平。分析RA组血清MCP-4水平是否与ESR、CRP、RF及双手放射学检查等指标具有相关性。结果:RA组血清MCP-4水平为(203.79±18.64)pg/mL,其它自身免疫性疾病对照组血清MCP-4水平为(207.76±40.37)pg/mL,正常健康对照组血清MCP-4水平为(125.13±11.08)pg/mL。RA组、其它自身免疫性疾病对照组血清MCP-4水平与正常健康对照组相比均有统计学意义(Plt;0.05),RA组与其它自身免疫性疾病对照组间无统计学意义(P=0.787),RA活动期患者血清MCP-4水平(214.86±24.46)pg/mL和非活动期患者血清MCP-4水平(190.27±29.11)pg/mL比较无统计学意义(P=0.065)。RA组血清MCP-4水平与ESR、CRP、RF及双手X片分期无相关性。结论:RA组的血清MCP-4水平高于正常健康对照组,但与ESR、CRP、RF及双手X片分期无相关性。MCP-4可能参与了RA的发病过程。
Objective To investigate the effect of monocyte count to high density lipoprotein ratio (MHR) on early complications after off-pump coronary artery bypass grafting and to explore the predictive factors for early complications in patients after off-pump coronary artery bypass grafting. Methods The clinical data of patients who underwent simple off-pump coronary artery bypass grafting from October 2021 to September 2023 in our hospital were retrospectively analyzed. The patients were divided into a low value group and a high value group according to the median MHR value. The clinical data of the two groups were compared, and binary logistic regression analysis was used to explore the and predictors of atrial fibrillation (AF) and acute kidney injury (AKI) after coronary artery bypass grafting. Results A total of 220 patients were included, with a median MHR of 0.48. There were 108 patients in the low value group (MHR<0.48), including 71 males and 37 females, with an average age of 65.28±7.85 years. There were 112 patients in the high-value group (MHR≥0.48), including 84 males and 28 females, with an average age of 64.57±8.75 years. There was no statistical difference between the two groups in terms of general basic data such as gender or age (P>0.05). The incidence of postoperative AF and AKI in the high-value group was significantly higher than that in the low-value group (P<0.05), and no statistical difference in terms of other postoperative complications was observed. Binary logistic regression analysis showed that MHR was a risk factor for postoperative AKI and postoperative AF (P<0.05). Conclusion The study shows that MHR is a risk factor for new-onset AF and AKI after coronary artery bypass grafting.
Objective To investigate the effect of monocyte chemoattractant protein 1 (MCP-1) on the migration of the induced and differentiated mouse bone marrow mesenchymal stem cells (BMSCs) for raising the efficacy of intravenous transplantation of BMSCs. Methods The BMSCs were cultured with the method of differential adhesion and density gradient centrifugation of C57/BL10 mice, and were identified by alkal ine phosphatase Gomori modified staining after osteogenic inducing. At the 3rd passage, the BMSCs were induced to the myoblasts with 5-azacytidine (5-Aza). The chemotaxis of MCP-1 in the induced and differentiated BMSCs in vitro at concentrations of 25, 50, 100, 200, and 400 ng/mL was observed through the migration test, by counting the number of the migrated cells. The expression of the chemokine receptor 2 (CKR-2) in the induced and differentiated BMSCs was detected with the flow cytometry. Results The cells could be cultured with the methods of differential adhesion and density gradient centrifugation and still had higher prol iferative and differentiative potency; the induced cells at the 3rd passage could differenciate to the osteoblasts, confirming that the cells were BMSCs; the myogenic induced BMSCs possesed the sarcotubule structure. The number of the migrating BMSCs at MCP-1 concentrations of 25-400 ng/ mL were respectively 35.066 7 ± 6.584 2, 43.200 0 ± 6.460 8, 44.466 7 ± 4.823 5, 45.600 0 ± 8.650 3, and 50.733 3 ± 7.582 5; showing significant difference when compared with control group (28.333 3 ± 8.917 6, P lt; 0.05), and presenting significant difference among 25, 50, 400 ng/mL groups compared with each other (P lt; 0.05). The expression of CKR-2 in the mouse BMSCs (48.0%) was significantly higher (P lt; 0.001) than those of blank control (0.6%) and negative control (17.0%). Conclusion The results indicate that the MCP-1 can induce the migration of mouse BMSCs by MCP-1/CKR-2 pathway.
ObjectiveTo investigate the expression and significance of cysteine-rich protein 61 (Cyr61) in patients with chronic obstructive pulmonary disease (COPD).MethodsBetween September 2017 and September 2018, 27 patients with benign tumor needing to surgical therapy, were divided into COPD group (15 patients) and non-COPD group (12 patients), according to lung function. Lung tissues were selected at the distance at least 5 cm from the tumor. The levels of Cyr61, interleukin-8 (IL-8) and monocyte chemoattractant protein-1 (MCP-1) in serum were determined by enzyme-linked immunosorbent assay. Meanwhile, the expressions of Cyr61 in lung tissues were measured by immunohistochemistry technology between two groups. Furthermore, correlations among Cyr61, IL-8, MCP-1, smoking index, forced expiratory volume in the 1st second as percentage of predicted values (FEV1%pred), scores of COPD Assessment Test (CAT) were analyzed.ResultsSerum Cyr61, IL-8, MCP-1 levels were significantly higher in patients with COPD than in the non-COPD group (P<0.05), (2409.80±893.87)pg/mL, (76.27±10.53)pg/mL, (173.67±42.64)pg/mL vs. (1065.42±158.83)pg/mL, (57.33±8.29)pg/mL, (138.42±27.62)pg/mL, respectively. By immunohistochemistry technology, the expression levels of Cyr61 in lung epithelial cells and in lung macrophage cells of COPD patients were higher than in the non-COPD group (P<0.01). Positive correlations were found between serum IL-8, serum MCP-1, CAT scores, smoking index and serum Cyr61 (r=0.674, 0.566, 0.602, and 0.755, P=0.006, 0.028, 0.018, and 0.003, respectively) in COPD group. Furthermore, in COPD group, there were also positive correlations between serum IL-8, serum MCP-1, CAT scores, smoking index and intrapulmonary Cyr61 (r=0.542, 0.635, 0.809, and 0.580, P=0.037, 0.011, 0.001, and 0.038 respectively). Inverse correlation was found between serum Cyr61 and FEV1%pred (r=–0.772, P<0.01), and the same as between intrapulmonary Cyr61 and FEV1%pred (r=–0.683, P<0.01).ConclusionsCyr61 highly expresses in serum and in lung tissues of patients with COPD, and its expression is correlated with lung function of patients. The results indicate that Cyr61 may interact with IL-8 and MCP-1 in the pathogenesis of chronic obstructive pulmonary disease.
ObjectiveTo explore the expression of programmed death-1 ligand (PD-L1) on peripheral blood monocytes in septic mice, and analyze the difference between the mild and severe septic mice. MethodsFirstly, thirty C57 mice were randomly divided into a sham group, a mild sepsis group and a severe sepsis group. Sepsis model was induced by cecal ligation and puncture (CLP). The severity of sepsis was distinguished by length of cecum ligated. Survival rate was recorded after CLP and compared between three groups. Then sixty C57 mice were randomly divided into a sham group, a mild sepsis group and a severe sepsis group. Peripheral blood was obtained at 6 h and 24 h to detect PD-L1 expression on monocytes in peripheral blood by flow cytometry. Tumor necrosis factor-α(TNF-α) and interleukin-10 (IL-10) in serum were detected by enzyme-linked immunosorbent assay. ResultsThe survival rate in the mild sepsis group was higher than that in the severe sepsis group. TNF-αand IL-10 levels in the mild and severe sepsis groups were higher than those in the sham group at 6 h and 24 h (P < 0.01). PD-L1 expression on monocytes in the mild and severe sepsis groups was higher than that in the sham group at 6 h and 24 h (P < 0.05). The expression of PD-L1 in the severe sepsis group was higher than that in the mild sepsis group, however, there was no statistical difference between two groups (P > 0.05). ConclusionsPD-L1 expression on monocytes is increased in septic mice. PD-L1 expression tended to increase in severe sepsis compared with the mild sepsis.
【摘要】 目的 探讨儿童传染性单核细胞增多症（IM）的临床特点。 方法 回顾性分析2005年8月-2009年8月收治的151例IM患儿的症状、体征、实验室检查及治疗效果。 结果 IM临床表现以发热、咽峡炎、肝脾和淋巴结肿大最常见，鼻塞、眼睑浮肿也是重要体征。外周血出现异型淋巴细胞及EBV-IgM抗体检测可帮助确诊。更昔洛韦治疗IM效果确切。 结论 应重视IM临床特点，有助于早期诊断并提高确诊率，应用更昔洛韦治疗值得推广。【Abstract】 Objective To investigate the clinical features of infectious mononucleosis (IM) in children. Methods A total of 151 children with infectious mononucleosis admitted from August 2005 to August 2009 were reviewed. The symptoms, signs, laboratory tests, and treatment of infectious mononucleosis were analyzed. Results Fever, angina, hepatomegaly, splenomegaly, and swollen lymph nodes were the most common clinical manifestations of IM. Nasal congestion and eyelid edema were also two important signs. Atypical lymphocytes in peripheral blood and EBV antibody (VCA-IgM) could help confirm the diagnosis. The antiviral treatment with ganciclovir was effective for infectious mononucleosis. Conclusion Clinical features of infectious mononucleosis are helpful to the diagnosis. Treatment with ganciclovir should be promoted.
ObjectiveTo investigate the role of Krüppel-like factor 4 (KLF4) mediated monocyte/macrophage subtype switch in the pathological progression of pulmonary fibrosis.MethodsThirty-six patients with interstitial pneumonia were recruited from Characteristic Medical Center of the Chinese People's Armed Police Force between May 2015 and January 2017. Peripheral venous blood and bronchoalveolar lavage fluid were collected in the morning. Pulmonary function and arterial blood gas were tested after admission. Flow cytometry was used to test monocyte subtypes of peripheral blood and macrophage subtypes of bronchoalveolar lavage fluid. KLF4 of peripheral blood was detected by enzyme linked immunosorbent assay. Thirty normal subjects were selected as control group of peripheral blood mononuclear cell subtypes and KLF4 (control group A), and 10 patients without pulmonary fibrosis who needed bronchoscopy were selected as control group of macrophage subtypes in alveolar lavage fluid (control group B). The relationship between the expression of KLF4 and the differentiation of monocytes and macrophages were observed. Furthermore, the relationship between the differentiation of monocytes subtypes, macrophages subtypes and lung function were observed.ResultsMonocyte of CD14++CD16– subtype in pulmonary fibrosis group was significantly lower than that in control group A (P<0.05). Monocyte of CD14++CD16+ subtype in pulmonary fibrosis group was significantly higher than that in control group A (P<0.05). No significant difference was found between the two groups regarding CD14+CD16++. No correlation was found between three subtypes of monocyte and DLCO of patients and between three subtypes of monocyte and PaO2 of patients. M1 macrophage in pulmonary fibrosis group was significantly lower than that in control group B (P<0.05). M2 macrophage in pulmonary fibrosis group was significantly higher than that in control group B (P<0.05). Negative correlation was found between the ratio of M2 subtypes and DLCO of patients and between the ratio of M2 subtypes and PaO2 of patients (P<0.05). KLF4 protein of blood in pulmonary fibrosis group was significantly higher than that in control group A (P<0.05). Positive correlation was found between the ratio of M2 subtypes and KLF4 protein (P<0.05).ConclusionsCD16+ monocyte plays a role in the occurrence and development of pulmonary fibrosis, but no evidence is found there is a direct correlation between monocyte subtypes of peripheral blood and fibrosis degree of lung tissue. M2 macrophage subtype plays an important role in the development of interstitial pneumonia. The number of M2 macrophages is positively correlated with the severity of pulmonary fibrosis. Monocyte/macrophage subtype differentiation by KLF4 may play a role in the pathological progression of pulmonary fibrosis.
ObjectiveTo observe the effects and mechanism of MCP-1 in ileum and pancreatic tissues in rats with severe acute pancreatitis(SAP). MethodsTwenty-fourth healthy SD rats were randomly divided into two groups:control group(n=12) and SAP model group(n=12). SAP was induced in model group by retrograde injection of 3% sodium taucrocholate into the biliopancreatic duct of rats. The control group underwent laparotomy with the manipulation of the intestinal canal. The rats were killed at 12 h and 24 h respectively after operation, blood and tissue samples were collected to detect the indexes as follows:①Expressions of MCP-1 mRNA of pancreatic and ileum tissues were detected by RT-PCR; ②blood plasma MCP-1 and IL-10 levels were detected by ELISA; ③blood plasma AMY and DAO levels were detected by colorimetry; ④the pathological changes of pancreas and ileum tissues were observed. ResultsCompared with the control group, the levels of MCP-1, IL-10, AMY, and DAO in plasma, pancreas, and ileum tissues were significantly increased in SAP model group(P < 0.01), the expressions of MCP-1 mRNA in pancreas and ileum tissues were up-regulated simultaneously(P < 0.01), and pathological scoring increased obviously(P < 0.01). ConclusionThe levels of MCP-1 in plasma, pancreas and ileum tissues are significantly increased in rats with SAP, MCP-1 aggravate the injury of pancreas and ileum tissues.