ObjectiveTo explore the expression of senescence marker protein-30 (SMP-30) in human lung tissues and the significance in the pathogenesis of chronic obstructive pulmonary disease (COPD). MethodsLung tissue specimens ( > 5 cm away from cancerous tissues) obtained by surgery resection in 20 subjects with solitary peripheral carcinoma in Jiangsu Province Hospital were investagted. The subjects were divided into three groups according to lung function and smoking history, ie. a COPD group (6 cases), a healthy smoking group (7 cases) and a healthy control group (7 cases). Immunohistochemistry and Western blot were used to determine the distribution and expression of SMP-30 in human lung tissues. ResultsSMP-30 protein mainly expressed in the cytoplasm of alveolar macrophages (AM). The numbers of AM and SMP-30-positive AM were significantly increased in the COPD group. Western blot analysis confirmed a significant increase in SMP-30 expression in the healthy smokers compared with the non-smokers (2.16±0.23 vs. 1.10±0.14, P < 0.01) and further enhanced in the patients with COPD compared with the healthy smoking subjects (4.62±0.97 vs. 2.16±0.23, P < 0.05). The levels of the protein in different groups were: COPD group > smoking group > control group with significant difference. ConclusionThese results suggest that SMP-30 expression may be involved in the mechanism of prolonged survival and the increase in number of AM and may be involved in the pathogenesis of COPD.
ObjectiveTo investigate the changes of peritoneal macrophages function of mice with gastric cancer in the CO2 pneumoperitoneum environment, as well as its effect on the peritoneal metastasis of gastric cancer. MethodsAn orthotopic implantation model of mouse forestomach cancer was established using the 615 mouse. The mice bearing tumors were randomly divided into five groups (n=30): anesthesia alone, laparotomy, and 2, 4, and 6 mm Hg CO2 insufflation groups. Peritoneal macrophages were collected from six mice in each group and cultured. The macrophage phagocytic function on neutral red and the levels of NO and TNF-α produced by macrophages were measured after 12, 24, 48, and 72 h of culture. The remaining mice were observed after two weeks for the rate of peritoneal metastasis of forestomach cancer cells and the total weight of implanted nodules. ResultsNo death and ascites were found and the difference of weight body was not significant in all mice (Pgt;0.05). The uptake of neutral red by peritoneal macrophages and the levels of NO and TNF-α secreted by peritoneal macrophages in the laparotomy group after 12 h of culture were all significantly higher than those in other four groups (Plt;0.05). The corresponding values in the 2, 4， and 6 mm Hg CO2 insufflation groups after 12 h were all significantly lower than those in the anesthesia alone group (Plt;0.05). Among three insufflation groups, the corresponding values in the 2 mm Hg after 12 h were significantly higher than those in the 4 and 6 mm Hg CO2 insufflation group, though the difference in the two latter was not significant (Pgt;0.05). The uptake of neutral red by peritoneal macrophages and the levels of NO and TNF-α secreted by peritoneal macrophages in the 6 mm Hg CO2 insufflation group after 24 h of culture were all significantly lower than those in other four groups (Plt;0.05), while the difference in the four groups was not significant (Pgt;0.05). The uptake of neutral red by peritoneal macrophages and the levels of NO and TNF-α secreted by peritoneal macrophages after 48 h and 72 h were not significantly different in the five groups (Pgt;0.05). The rate of peritoneal metastasis of mice was significantly lower in the 6 mm Hg insufflation CO2 group (75.0%, 15/20) than that in the anesthesia alone group (100%, 24/24), Plt;0.05, but higher than other three groups(Plt;0.05), which was not different in 2 mm Hg (47.8%, 11/23), 4 mm Hg insufflation group (45.45%, 10/22) and laparotomy group (50.0%, 10/20), Pgt;0.05. The total weight of implanted nodules of mouse forestomach cancer was (1.24±0.48) g, (1.02±0.38) g, (0.96±0.33) g, (0.93±0.45) g, and (1.18±0.37) g in the anesthesia alone group, the laparotomy group, and 2, 4, and 6 mm Hg CO2 insufflation group, and the difference was not significant (Pgt;0.05). ConclusionHigh pressure (6 mm Hg) CO2 pneumoperitoneum can constantly inhibit the phagocytosis and cytokine secretion functions of peritoneal macrophages in gastric cancer-bearing mice and promote peritoneal implantation of gastric cancer.
ObjectiveTo describe the clinical,radiographic,and laboratory features of autoimmune pulmonary alveolar proteinosis (PAP) from a single center. MethodsConsecutive autoimmune PAP cases diagnosed in the Nanjing Drum Tower Hospital between January 2006 and December 2012 were recruited in the study. The clinical,radiographic and laboratory data of the PAP patients were analyzed to explore the clinical significance of serum GM-CSF autoantibody (GMAb) and serum cytokeratin (CYFRA21-1). ResultsThe median serum GMAb level of the 26 cases was 28.64 μg/mL (interquartile range,19.2-75.4 μg/mL),which were diagnosed as autoimmune PAP based on the serum GMAb levels of these patients all above the cut-off value of 2.39 μg/mL while the serum GMAb levels of 30 normal controls were 0.10(0.05-0.15)μg/mL and all below the cut-off value. 34.6% of all recruited 26 autoimmune PAP patients had identified occupational inhalational exposure. There was no significant correlation in the serum GMAb in autoimmune PAP patients with disease severity scores (DSS),lung function parameters,chest high resolution computed tomography (HRCT) scores,or PaO2 (P>0.05). There was significant correlation of DSS of autoimmune PAP patients with PaO2,FVC%pred,TLCO%pred,opacity extent score of chest HRCT,and opacity severity score of chest HRCT (P<0.05). The median serum level of CYFRA21-1 of the autoimmune PAP patients was 9.9(4.3-19.5)ng/mL,which was significant higher than that of the normal control group (P<0.05). However there was no significant correlation in the serum CYFRA21-1 in the autoimmune PAP patients with DSS,lung function parameters,and chest HRCT scores. 92.3% of the chest HRCT of 26 autoimmune PAP patients had crazy paving sign,while 100% of them had geographic sparing sign. ConclusionSerum GMAb and CYFRA21-1 may be important biomarkers for diagnosis of autoimmune PAP. The PAP with occupational inhalational exposure constitutes a high proportion of autoimmune PAP patients.
ObjectiveTo establish a methodology for alveolar macrophages (AMs) phagocytosis of AlexaFluor 488 (AF488) labeled bacteria by flow cytometry.MethodsStaphylococcus aureus and Streptococcus pneumoniae were labeled with different concentrations of AF488. A flow cytometric assay was used to quantify in vivo bacterial uptake by AMs. AMs and different ratio of fluorescent-labeled bacteria were incubated at 37 ℃ for 2 hours, 4 hours, 6 hours and 8 hours, respectively. AMs were washed with DPBS and extracellular fluorescence was quenched with 1% (w/v) trypan blue. Trypan blue was aspirated and phagocytosis of fluorescent-labeled bacteria by AMs was measured using a flow cytometry. Confocal microscopy was performed to ensure that bacterial in positive AM had been internalized rather than bound to the cell surface.ResultsWhen the concentration of AF488 was more than 50 μg/mL, the labeling rates of Staphylococcus aureus and Streptococcus pneumoniae were higher than 92% (P<0.05), and has quickly reached the upper limit. With the prolongation of incubation time, the phagocytic rate of AMs increased from 20.4% at 2 hours to 76.5% at 8 hours. With the increase in the number of bacteria, the phagocytic rate of AMs increased from 7.7% by ratio of 1∶10 to 85.1% by ratio of 1∶300.ConclusionDetection of AMs phagocytosis of AF488 labeled bacteria by flow cytometry is an effective method, but the dye concentration, incubation time and the proportion of bacteria will influence the results.
Acute respiratory distress syndrome (ARDS) is the most common cause of acute respiratory failure. Extensive researches have been conducted for the pathophysiology of this disease, but the mortality rate remains high. Previous studies have found that catecholamines play an important role in acute lung injury, and newly discover prompted that upregulation of phagocyte-derived catecholamines augmented the acute inflammatory response in acute lung injury which provides a new way of thinking. In the current review, we describe the mechanism of the phagocyte-derived catecholamines augmenting the acute lung injury.
Objective To investigate the evaluation value of serum interleukin-34 (IL-34), macrophage migration inhibitor (MIF), osteopontin (OPN) and hypersensitive C-reactive protein (hs-CRP) in the diagnosis and prognosis of active pulmonary tuberculosis. Methods Clinical data of 100 patients with active pulmonary tuberculosis admitted from June 2019 to June 2022 were selected as an observation group and retrospectively analyzed. All patients received standardized anti-tuberculosis therapy for 6 months and were divided into a good prognosis group (76 cases) and a poor prognosis group (24 cases) according to the prognosis. Another 80 healthy volunteers who underwent physical examination during the same period were selected as the control group. Serum levels of IL-34, MIF, OPN and hs-CRP were detected in each group, and the value of serum IL-34, MIF, OPN and hs-CRP in the diagnosis and prognosis of active pulmonary tuberculosis was analyzed by receiver operating characteristic curve (ROC curve). Results Serum levels of IL-34, MIF, OPN and hs-CRP in the observation group were higher than those in the control group (all P<0.05). ROC curve showed that serum IL-34, MIF, OPN, hs-CRP had a certain diagnostic value in active pulmonary tuberculosis, with area under ROC curve (AUC) of 0.864, 0.870, 0.865, and 0.880, respectively (all P<0.01), and the combination of the four indexes had a higher diagnostic value (AUC=0.902, P<0.01). Serum levels of IL-34, MIF, OPN and hs-CRP in the good prognosis group were lower than those in the poor prognosis group (all P<0.05). ROC curve showed that serum IL-34, MIF, OPN, hs-CRP had a certain value in evaluating the prognosis of active pulmonary tuberculosis, with AUC of 0.850, 0.874, 0.837, and 0.842, respectively (all P<0.01), and the combined value of the four indexes was higher (AUC=0.923, P<0.01). Conclusion The combined detection of serum IL-34, MIF, OPN and hs-CRP has high value in the diagnosis and prognosis assessment of active pulmonary tuberculosis.
ObjectiveTo construction the telmisartan/collagen/polycaprolactone (PCL) nerve conduit and assess its effect on repairing sciatic nerve defect in rats. Methods The 60% collagen/hexafluoroisopropanol (HFIP) solution and 40% PCL/HFIP solution were prepared and mixed (collagen/PCL solution). Then the 0, 5, 10, and 20 mg of telmisartan were mixed with the 10 mL collagen/PCL solution, respectively. Telmisartan/collagen/PCL nerve conduits were fabricated via high voltage electrospinning technology. The structure of nerve conduit before and after crosslinking was observed by using scanning electron microscope (SEM). The drug release efficiency was detected by in vitro sustained release method. RAW264.7 cells were cultured with lipopolysaccharide to induce inflammation, and then co-cultured with nerve conduits loaded with different concentrations of telmisartan for 24 hours. The mRNA expressions of inducible nitric oxide synthase (iNOS) and Arginase 1 (Arg-1) were detected by using real-time fluorescence quantitative PCR. Forty adult Wistar rats were randomly divided into 4 groups (n=10). After preparing 15-mm-long sciatic nerve defect, the defect was repaired by cross-linked nerve conduits loaded with 0, 5, 10, and 20 mg telmisartan in groups A, B, C, and D, respectively. After operation, the general condition of rats was observed after operation; the sciatic function index (SFI) was tested; the bridging between the nerve conduit and sciatic nerve, and the integrity of nerve conduit were observed; the tissue growth in nerve conduit and material degradation were observed by HE staining; the expressions of CD86 (M1 macrophage marker), CD206 (M2 macrophage marker), myelin basic protein (MBP), and myelin protein 0 (P0) in new tissues were also observed by immunohistochemical staining; the expressions of neurofilament 200 (NF-200) and S-100β in new tissues were assessed by immunofluorescence staining. Results The general observation showed that the inner diameter of the nerve conduit was 1.8 mm and the outer diameter was 2.0 mm. After cross-linking by genipin, the nanofiber became thicker and denser. The drug release test showed that the telmisartan loaded nerve conduit could be released gradually. With the increase of telmisartan content in nerve conduit, the iNOS mRNA expression decreased and the Arg-1 mRNA expression increased; and the differences between 20 mg group and other groups were significant (P<0.05). In vivo experiment showed that all animals in each group survived until the completion of the experiment. The SFI was significantly higher in groups C and D than in groups A and B at different time points (P<0.05) and in group D than in group C at 6 months after operation (P<0.05). HE staining showed that there were significantly more new tissues in the middle of the nerve conduit in group D after operation than in other groups. Immunohistochemical staining showed that CD86 and CD206 stainings were positive in each group at 1 month after operation, among which group D had the lowest positive rate of CD86 and the highest positive rate of CD206, and there were significant differences in the positive rate of CD206 between group D and groups A, B, and C (P<0.05); the MBP and P0 stainings were positive in groups C and D at 6 months, and the positive rate in group D was significantly higher than that in group C (P<0.05). Immunofluorescence staining showed that the NF-200 and S-100β expressions in group D were significantly higher than those in other groups. ConclusionTelmisartan/collagen/PLC nerve conduit can promote the sciatic nerve defect repair in rats through promoting the polarization of M1 macrophages to M2 macrophages, and the nerve conduit loaded with20 mg telmisartan has the most significant effect.
ObjectiveTo investigate the effect of curcumin on lipopolysaccharide (LPS)-induced inflammation and apoptosis in alveolar macrophage via microRNA-132 (miR-132)/high mobility group protein B1 (HMGB1).MethodsThe cultured mouse alveolar macrophage line (RAW264.7 cells) were divided into the control group, the LPS group, the LPS+50 μmol/L curcumin group, and the LPS+100 μmol/L curcumin group. Forty-eight hours after drug treatment, the levels of miR-132/HMGB1, inflammatory mediator and apoptotic were detected. Secondly, the empty vector, synthetic miR-132 mimics and inhibitors were transfected into another cultured mouse alveolar macrophage line (RAW264.7 cells) to detect the inflammation and apoptosis of alveolar macrophage after transfection.ResultsCompared with the control group, in the LPS group, the apoptosis of alveolar macrophage, the levels of interleukin (IL)-6, IL-8 and tumor necrosis factor (TNF)-α, and the expression of miR-132 increased, while the expression of HMGB1 decreased (P<0.05); compared with the LPS group, in the two curcumin groups, the apoptosis of alveolar macrophage, the levels of IL-6, IL-8 and TNF-α, and the expression of miR-132 decreased, while the expression of HMGB1 increased (P<0.05); and the greater the drug concentration, the more obvious the effect (P<0.05). In addition, up-regulation of miR-132 reduced the expression of HMGB1 in alveolar macrophage, increased inflammatory factor, and induced apoptosis in alveolar macrophage; however, down-regulation of miR-132 increased the expression of HMGB1 in alveolar macrophage, reduced inflammatory factor, and inhibited apoptosis in alveolar macrophage (P<0.05).ConclusionCurcumin could decrease LPS-induced inflammation and apoptosis in alveolar macrophage via decreasing miR-132 and increasing HMGB1.
Objective To observe the effect of transfer of immature mouse myeloid dendritic cells （DC） generated with low-dose granulocyte-macrophage colony-stimulating factor （GM-CSF） on cardiac allograft survival. Methods Mouse DC were generated with standard doses or low doses GM-CSF from bone marrow cells, the phenotype and functional properties of these DC were compared through fluorescence-activated cell sorting（FACS） analysis and mixed lymphocyte reaction（MLR）, 1. 0 × 106 DC generated with low doses GM-CSF were administered to the recipients 7 days before transplantation, and the cardiac allograft survival were observed. Results In contrast to DC generated with standard doses, DC generated with low doses were phenotypically immature DC （CD11c＋, CD80- , CD86- , MHCⅡlow）, and induced allogeneic T cell unresponsiveness, and administration of these DC to recipients prolonged cardiac allograft survival from 6.3±1.2 days to 14.3±1.9 days. Conclusions DC generated from mouse bone marrow progenitors in low doses of GM-CSF are phenotypically and functionally immature, and prolong cardiac allograft survival when they are administered 7 clays before transplantation.
Objective To summarize the research progress on the role of macrophage-mediated osteoimmune in osteonecrosis of the femoral head (ONFH) and its mechanisms. Methods Recent studies on the role and mechanism of macrophage-mediated osteoimmune in ONFH at home and abroad were extensively reviewed. The classification and function of macrophages were summarized, the osteoimmune regulation of macrophages on chronic inflammation in ONFH was summarized, and the pathophysiological mechanism of osteonecrosis was expounded from the perspective of osteoimmune, which provided new ideas for the treatment of ONFH. Results Macrophages are important immune cells involved in inflammatory response, which can differentiate into classically activated type (M1) and alternatively activated type (M2), and play specific functions to participate in and regulate the physiological and pathological processes of the body. Studies have shown that bone immune imbalance mediated by macrophages can cause local chronic inflammation and lead to the occurrence and development of ONFH. Therefore, regulating macrophage polarization is a potential ONFH treatment strategy. In chronic inflammatory microenvironment, inhibiting macrophage polarization to M1 can promote local inflammatory dissipation and effectively delay the progression of ONFH; regulating macrophage polarization to M2 can build a local osteoimmune microenvironment conducive to bone repair, which is helpful to necrotic tissue regeneration and repair to a certain extent. Conclusion At present, it has been confirmed that macrophage-mediated chronic inflammatory immune microenvironment is an important mechanism for the occurrence and development of ONFH. It is necessary to study the subtypes of immune cells in ONFH, the interaction between immune cells and macrophages, and the interaction between various immune cells and macrophages, which is beneficial to the development of potential therapeutic methods for ONFH.