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Objective:To analyze the differentially expressed gene (DEG) in rats with sepsis-induced exogenous acute respiratory distress syndrome (ARDS) and explore the early diagnosis and protective mechanism of sepsis-induced ARDS at the transcriptome level.Methods:Twelve 6 to 8 weeks old male Sprague-Dawley (SD) rats were randomly divided into lipopolysaccharide (LPS) induced sepsis-induced ARDS model group (model group, intraperitoneal injection of LPS 15 mg/kg) and control group (intraperitoneal injection of the same volume of normal saline), with 6 rats in each group. RNA was extracted from the left lung tissue of the two groups, and the paired-end sequencing mode of the illumina Hiseq sequencing platform was used for high-throughput sequencing. The DESeq2 software was used to screen DEG with | log 2 (fold change, FC) | ≥ 3 and P < 0.001. Gene ontology (GO) function enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were performed on DEG. STRING and CytoScape software were used to construct a protein-protein interaction (PPI) network and screen key genes. The peripheral blood mononuclear cell (PBMC) of 20 septic patients admitted to the emergency and critical care medical department of Lianyungang First People's Hospital from March to November 2021 and 20 age-matched healthy people in the same period were isolated and extracted, and the key genes were verified by real-time fluorescent quantitative polymerase chain reaction (RT-qPCR). Results:A total of 286 DEG were screened, including 202 up-regulated genes and 84 down-regulated genes. GO enrichment analysis showed that DEG was mainly involved in biological processes such as neutrophil chemotaxis migration, antibacterial humoral response, host immune response, and humoral immune response. KEGG analysis showed that DEG mainly played a biological role through interleukin-17 (IL-17) signaling pathway, tumor necrosis factor (TNF) signaling pathway, and chemokine signaling pathway. In PPI analysis, a total of 262 node proteins were screened, and the interaction relationship was 852 edges. The first 15 key genes were IL-6, TNF, IL-10, IL-1β, chemokine ligand 1 (CXCL1), CXCL10, chemokine receptor 3 (CXCR3), CXCR2, CXCL9, chemokine ligand 7 (CCL7), CXCL11, CCL1, CXCL13, CCL12, and CCL22. Five representative key genes were performed on PBMC of blood samples from septic ARDS patients and healthy controls by RT-qPCR. The results showed that their expression was significantly higher than that in the healthy controls [IL-6 mRNA (2 -ΔΔCt): 2.803±1.081 vs. 0.951±0.359, TNF mRNA (2 -ΔΔCt): 2.376±0.799 vs. 1.150±0.504, CXCL10 mRNA (2 -ΔΔCt): 2.500±0.815 vs. 1.107±0.515, CXCR3 mRNA (2 -ΔΔCt): 1.655±0.628 vs. 0.720±0.388, CCL22 mRNA (2 -ΔΔCt): 1.804±0.878 vs. 1.010±0.850, all P < 0.05], and the trends were consistent with the RNA-Seq results. Conclusion:Biological processes such as chemotactic migration and degranulation of inflammatory cells, cytokine immune response, and signal pathways such as CXCL10/CXCR3 and IL-17 play important roles in the occurrence and development of sepsis-related exogenous ARDS, which would provide new ideas and targets for further study of lung injury mechanisms and clinical prevention and treatment.
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Objective To evaluate the efficacy and safety of locating the target bronchus and treating through the chest wall and airway intervention in 29 cases with lung abscess secondary to pulmonary bulla. Methods In 29 cases the lung abscess secondary to pulmonary bulla had a mean diameter of 14 cm, of which 25 with COPD. Implement-ing 4 different ways to locate the target bronchus of lung abscess: Preoperative pulmonary CT, conventional fiber bronchoscopy, and locating the target bronchus by infecting colored saline separately forward and backward. Under guiding of CT, the central venous catheter was inserted via chest wall into lung abscess, which had pus suction, lavage, medicine injection and continuous negative pressure suction and then under direct vision of bronchoscopy, all of the target bronchi of pulmonary absess were occluded with fibrin glue. Results All of the 29 cases, the target bronchi of lung abscess were localized by lung CT in 17, by conventional bronchoscopy in 23, and by infecting col-ored saline forward in 26 and backward in 24 respectively. Cured in 25 and improved in 3, the total effective rate was 96.6%. The adverse effects:chest pain in 3, pneumothorax in 2, pyothorax, pyopneumothorax, subcutaneous em-physema, the drainage catheter falling off and dislocating separately in 1. Conclusion It was simple, safe and effec-tive to treat lung abscess secondary to lung bulla through the chest wall and airway intervention.
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Objective: To identify the relationship between the expression of protein tyrosine phosphatase non-receptor type 12 (PTPN12) and radiotherapy effect in non-small cell lung cancer (NSCLC) tissues and to determine whether PTPN12 deficiency can sensi-tize lung cancer cells to irradiation. Methods: From September 2013 to October 2014, 92 NSCLC patients undergoing radiotherapy with or without platinum-based combination chemotherapy were analyzed retrospectively. Before the treatment, PTPN12 expression was detected through immunohistochemistry. After the completion of radiotherapy, the patients' responses were assessed and radio-therapeutic efficacy analyzed. The human NSCLC cell line H1299 was infected with shPTPN12 knockdown, and colony survival assay was analyzed after irradiation. Chi-square test was used to examine the correlation between PTPN12 expression and clinicopathologi-cal characteristics. Univariate analyses and Logistic regression test were used to analyze the relationship between clinicopathological characteristics and radiotherapeutic response. Results: Patients with low PTPN12 expression were more sensitive to radiotherapy than those with high PTPN12 expression (80.0%vs. 57.1%, P=0.018). Multivariate analysis showed that PTPN12 expression was the on-ly independent predictor of radiotherapeutic response in NSCLC. The H1299-shPTPN12-knockdown cells were sensitive to irradiation. Conclusions:The results of the study indicated that downregulation of PTPN12 improved the radiosensitivity of NSCLC cells.
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Objective To investigate the expression of helper T cells (Th)17/regulatory T cells (Treg) balance associated factors in rheumatoid arthritis (RA) patients and their correlation with serum midkine (MK).Methods A total of 60 RA patients were divided into active RA patients (n=32) and inactive RA patients group (n=28).MK level in sera was detected by enzyme linked immunosorbent assay (ELISA) in 60 patients with RA and 30 healthy controls (HCs).The fraction of CD4+CD25+FOXP3+ Treg cells and IL-17+CD4+ Th17 cells in RA patients and healthy controls were determined by flow cytometry (FCM), and the expreasion of Foxp3, RORγt, Signal transducer and activator of transcription (STAT) 3 and STAT5 mRNA were detected with real-time polymerase chain reaction (PCR).Results were evaluated using ANOVA followed by q tests for comparisons of Th17 population between active RA patients, inactive RA patients and HCs, t test was used for comparing of Foxp3, RORγt, STAT3, STAT5 mRNA between RA group and HCs.The correlations between serum MK concentration and peripheral Treg cells, Th17 cells, Foxp3, RORγt, STAT3, STAT5 mRNA were analyzed by Pearson's correlation analysis.Results The percentages of Treg cells from active RA patients, inactive RA patients and HCs were significantly different (F=129.6, P＜0.01), the percentages of Treg cells of active RA patients [(1.41±1.05)％] were lower than that of the inactive RA patients [(3.6±1.6)％;q =7.92, P＜0.05] and healthy group [(7.7±1.7)％;q=22.45, P＜0.05], and there was significant difference between healthy group and inactive RA group (q=14.53, P＜0.05).The percentages of Th17 cells of the three groups were also significantly different (F=36.3, P＜0.01),the percentage of Th17 cells of active RA patients [(1.84±1.01)％] was significantly higher than that of inactive RA patients [(0.71±0.28)％;q=9.59, P＜0.05] and healthy group (0.53±0.16)％ [(P＜0.05;q=1 1.10, P＜0.05], there was no significant difference between the inactive RA group and healthy group (q=1.51, P＞0.05).The expression of RORγt and STAT3 mRNA in RA patients was higher than that of healthy controls (t=5.84, P＜0.01;t=4.52, P＜0.01).The expression of Foxp3 and STAT5 mRNA in RA patients were lower than healthy controls (t=6.01, P＜0.01;t=2.18, P＜0.05).Serum MK values were correlated with STAT5 (r=-0.55, P＜0.01), but not with Foxp3, RORγt, STAT3 mRNA or the percentage of Treg/Th17 cells.Conclusion Serum MK expression and the percentage of Th17 cells increase, while the percentage of Treg cells decrease in RA patients.Serum MK values are negatively correlated with STAT5 mRNA which is associated with Th17/Treg balance.This may be important in the pathogenesis of RA.
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Objective To investigate the relation between the apotosis of B cells in the peripheral blood (PB) and the expression of interleukin (IL)-17 in patients with rheumatoid arthritis (RA).Methods The proportions of apoptosis of B cells in the PB of 80 patients with RA and 80 healthy controls were measured by flow cytometry.B cells in the PB of 20 RA and 20 healthy individuals were isolated by MACS and Western blotting was used to detect the Bcl-2 and Caspase-3 protein levels.IL-17 levels were detected by enzyme-linked immunosorbent assay (ELISA).T-test and linear regression were used to analyze the data.Results The proportions of apoptosis of B cells in the PB of patients with RA and healthy controls were (14±6)％ and (24±9)％ respectively.The rate of apoptosis of B cells in patients with RA was significantly less than healthy controls (t=2.737,P=0.021).The Bcl-2 protein level of B cells in the PB of patients with RA group was significantly higher than that of control group (26±10,12±6,P＜0.01).Conversely,the Caspase-3 protein level of B cells in the PB of patients with RA group was significantly lower than that of the control group (16±7,31±12,P＜0.01).ELISA detected elevated level of serum IL-17 in the patients with RA as compared with controls [(69±19),(27±10) pg/ml,t=4.631,P=0.014].There was a negative correlation between the level of IL-17 and apoptosis of B cells in patients with RA (r=0.36,P＜0.01).Conclusion The elevated bcl-2 and reduced caspase-3 of B cells in patients with RA further proves there is abnormal apoptosis of B cells in RA patients.There is negative correlation between the expression of IL-17 and apoptosis of B cells in patients with RA and IL-17 can inhibit B cell apoptosis.