ObjectiveTo investigate the correlation between TLR5 rs5744174 gene polymorphism and Streptococcus pneumoniae pneumonia.MethodsOne-hundred and six patients with Streptococcus pneumoniae pneumonia admitted to this hospital from January 2014 to October 2018 were selected as an observation group, and 85 healthy subjects were selected as a control group during the same period. The clinical and pathological data of the subjects were collected, polymorphism of TLR5 rs5744174 gene was analyzed by PCR and sequencing, and the relationships between cell classification count, C-reactive protein (CRP) level in bronchoalveolar lavage fluid (BALF) and TLR5 rs5744174 gene polymorphism in the patients with Streptococcus pneumoniae pneumonia were analyzed.ResultsThere were significant differences in age, smoking, alcoholism, diabetes and the other general data between the observation group and the control group (P<0.05). The distribution of TLR5 rs5744174 genotype in the observation group and the control group was in accordance with Hardy-Weinberg equilibrium test level (χ2=16.89 for the observation group, χ2=10.76 for the control group, both P>0.05). There was no significant difference in the distribution frequency of TLR5 rs5744174 (C < T) genotype and allele between the two groups (P>0.05). There were significant differences in the proportion of diabetes mellitus among the three genotypes (CC, CT, TT) of the patients with Streptococcus pneumoniae pneumonia (P<0.05). The percentage of neutrophils and CRP levels in BALF were significantly different (P<0.05).ConclusionThe polymorphism of TLR5 rs5744174 gene may not be related to the occurrence of Streptococcus pneumoniae pneumonia, but is related to the proportion of complicated diabetes mellitus, the percentage of neutrophils and the level of CRP in patients with Streptococcus pneumoniae pneumonia, which may affect the degree of inflammation.
ObjectiveTo explore the effects of elafin on the regulation of mucin5AC (MUC5AC) in airway epithelial cells.MethodsAfter cultivating epithelial cells in vitro, cells were stimulated by cigarette smoke extract (CSE) and lipopolysaccharide (LPS), the intracellular MUC5AC production and extracellular secretion were assayed by immunofluorescence and ELISA, respectively.ResultsThe level of MUC5AC protein was obviously increased in the LPS stimulation group [(0.785±0.005) μg/ml vs. (0.232±0.004) μg/ml] and the CSE stimulation group [(0.803±0.005) μg/ml vs. (0.255±0.003) μg/ml] than those in the blank control groups (both P<0.01). When pretreated with elafin then stimulated by LPS and CSE, the MUC5AC productions were evidently decreased than those in the single LPS and CSE stimulation groups [(0.363±0.003) μg/ml and (0.406±0.006) μg/ml, bothP<0.01]. As compared with the blank control groups, the MUC5AC protein secretion were significantly decreased after stimulated with single elafin [(0.176±0.002) μg/ml and (0.193±0.004) μg/ml, bothP<0.05].ConclusionElafin may inhibit MUC5AC hypersecretion by blocking MUC5AC production and secretion.
ObjectiveTo detect the expressions of microvessel density (MVD)-CD34 and vascular endothelial growth factor (VEGF) in hepatic alveolar hydatid tissue in gerbil model and explore their clinical significances. MethodsSixty health gerbils were randomly equally divided into two groups, an experimental group and a sham operation group, each gerbil was given liver vaccination by opening their abdominal. Each gerbil in the experimental group was injected with approximately 400 echinococcus protoscoleces (0.1 mL), and each gerbil in the sham operation group received a corresponding volume of physiological saline. Six gerbils were sacrificed on day 20, 40, 60, 80, and 100. The hepatic alveolar hydatid tissue (AE) and its surrounding liver tissue (HSAE) were collected from the experimental group and the normal liver tissue (NL) was collected from the sham operation group, and the expressions of MVD-CD34 and VEGF were detected by immunohistochemistry staining (EnVision method). ResultsEchioncoccus multilocularis hydatid tissues were observed over the liver and in the partly abdominal cavity in the experimental group each gerbil by general observation. The expressions of CD34 and VEGF were observed in the AE at each time point after infection and located in the cytoplasmic of endothelial cells. The number of MVD-CD34 of AE at each time point in the AE was (9.83±3.87)/HP, (25.33±6.71)/HP, (34.50±5.50)/HP, (37.67±5.71)/HP and (44.67±4.93)/HP, respectively, which were significantly higher than those in the HSAE〔0/HP, (1.17±0.98)/HP, (3.50±1.38)/HP, (5.83±2.71)/HP, and(8.83±2.48)/HP, respectively〕and NL (all were 0), P < 0.05. The point of VEGF at each time point in the AE was 2.95±0.46, 3.90±0.68, 4.27±1.05, 5.33±0.95, and 4.50±0.81 respectively, which were significantly higher than those in the HSAE(1.07±0.63, 1.38±0.75, 1.55±0.83, 1.67±0.47, 2.10±0.55, respectively) and NL (1.02±0.83, 1.12±0.63, 1.26±0.26, 1.20±0.74, 1.21±0.28), P < 0.05. ConclusionAngiogenesis might be involved in infiltrated growth of alveococcus, and VEGF might contribute to angiogenesis of alveolar hydatid tissue.
ObjectiveTo investigate the synergistic effect of cold stress plus particulate matter 2.5 (PM2.5) co-exposure on the occurrence of respiratory inflammation and the possible post-transcriptional regulation mechanism of cold inducible RNA-binding protein (CIRP).MethodsIn vivo and in vitro experiments were carried out, and the lung tissue specimens from human surgical resection were observed. The rat model and cultured airway epithelial cells 16HBE were respectively divided into four groups (n=8), namely blank control group, 5 °C/18 °C group, PM2.5 group and 5 °C/18 °C+PM2.5 group. The expression of mRNA and protein of representative inflammatory cytokines and CIRP of cultured airway epithelial cells and rat bronchial/pulmonary tissues were respectively detected by ELISA, qPCR, and Western blot. Furthermore, the temporal dynamics of CIRP distribution were observed by cellular immunofluorescence. Finally, immunohistochemical method was used to observe the localization and expression of CIRP in rat and human bronchial/pulmonary tissues at the same time.ResultsIn vivo experiments, the mRNA and protein expression levels of CIRP, interleukin-6, and tumor necrosis factor-α in 5 °C group and PM2.5 group were significantly higher than those in the control group (all P<0.05), while the expression level of mRNA and protein in 5 °C+PM2.5 group were increased most obviously (all P<0.01). The same rule also appeared in the experimental results of each group in the vitro experiment. In addition, CIRP was mainly located in the cell nucleus; compared with the control group, the intracellular shift of CIRP appeared in 18 °C group and PM2.5 group, while the migration phenomenon was most obvious in the 18 °C+PM2.5 group. In the immunohistochemistry of rat bronchus/pulmonary tissue, the expressions of CIRP in the 5 °C group and in the PM2.5 group were significantly higher than those in the control group, and the CIRP expression in 5 °C+PM2.5 group was increased most evidently. Moreover, CIRP was expressed in the bronchial epithelial mucosa of normal people and patients with chronic obstructive respiratory disease (COPD), and it is mainly located in the nucleus of airway mucosal epithelial cells. The CIRP expression of COPD patients was significantly higher than that in the normal population.ConclusionCold stress has a sensitizing effect on airway epithelial inflammatory response induced by PM2.5, and post-transcriptional regulation of CIRP translocation from nucleus to cytoplasm may be an important mechanism.
Objective To investigate whether interleukin-1 receptor associated kinase (IRAK) participates the airway mucus hypersecretion induced by lipopolysaccharide (LPS). Methods The expression of Toll-like receptor (TLR)-4 or IRAK was down regulated by the transfection of specific small interfering RNA (siRNA). The transcription level of mucin 5AC (MUC5AC) mRNA as well as the secretion level of MUC5AC protein were judged by reverse transcription polymerase chain reaction (RT-PCR) and enzyme linked immunosorbent assay (ELISA), respectively. The activity of signaling molecules involved in TLR-4 associated pathway, such as the phosphorylated IRAK, nuclear translocation of NF-κB p65, was analyzed by Western blot. Results The level of intracellular phosphorylated IRAK was increased by stimulation of LPS in BEAS-2B cells. However, down-regulation of TLR-4 by siRNA could partially attenuate the additional phosphorylation of IRAK induced by LPS (P<0.05). LPS elicited a nuclear translocation of NF-κB p65 in BEAS-2B cells, which could be partially blocked by the down-regulation of TLR-4 or IRAK. With RT-PCR, an increased expression level of MUC5AC mRNA was discovered in wild-type BEAS-2B cells (0.82±0.09) than TLR-4 defect cells (0.36±0.05), IRAK defect cells (0.48±0.05) and IRAK inhibitor pretreated cells (0.57±0.07) (all P<0.05). Meanwhile, according to ELISA, an increased secretion level of MUC5AC protein was recorded in wild-type BEAS-2B cells [(0.76±0.09) μg/L] than TLR-4 defect cells [(0.33±0.04) μg/L], IRAK defect cells [(0.42±0.05) μg/L] and IRAK inhibitor pretreated cells [(0.51±0.06) μg/L] (all P<0.05). Conclusion As an crucial regulator of TLR dependent signaling pathway, IRAK may participate the airway mucus over-synthesis induced by LPS.