Transcriptome sequencing analysis of lncRNA expression in peripheral blood mononuclear cells from patients with COPD

Objective:To screen abnormally expressed lncRNAs in peripheral blood mononuclear cells from patients with COPD.Methods:The peripheral blood of 3 COPD patients and 3 normal controls were collected from our hospital,mononuclear cells were isolated,RNA was extracted and then transcriptome sequencing was performed.The expression difference between the two groups of samples was calculated based on p<0.05 and|log2FC|>1.Plot the difference lncRNA heat map and volcano map.The Lncpro database may predict mRNAs regulated by differential lncRNA,and perform the GO function and KEGG signaling clustering.Results:There were 67 lncRNAs between the COPD group and the control group that met the difference of p<0.05 and|log2FC|>1,of which 33 were up-regulated and 34 were down-regulated.Between the two groups.The target genes are mainly enriched in GO functions:regulatory functions of multicellular biological processes,regulatory functions of development processes,structured morphogenesis functions,system development functions,and development process functions.Target genes are mainly enriched in KEGG signaling pathways:multi-species apoptotic pathway,TGF-βsignaling pathway,complement and coagulation cascade pathway,colorectal cancer pathway and apoptosis pathway.Conclusion:Our results provide general information and possible regulatory functions and pathways of lncRNA expression changes in peripheral blood mononuclear cells of COPD,which may help clarify the underlying mechanism of COPD.

Expression and clinical significance of miR-186 in serum exosomes of patients with chronic obstructive pulmonary disease

Objective:To study the expression of miR-186 in serum exosomes of patients with chronic obstructive pulmonary disease,and to explore its clinical significance with chronic obstructive pulmonary disease.Methods:The peripheral blood sera of 53 COPD patients and 53 normal persons in our hospital were collected.Extraction of exosomes and electron microscopy to identify the characteristics of exosomes.RT-PCR method was used to detect the miR-186 expression level in serum exosomes.And statistical analysis of the expression difference between the COPD group and the normal group,alysis of the correlation between miR-186 and FEV1%predicted value,and ROC curve analysis of the diagnostic value of serum exosome miR-186 in COPD.Results:The relative expression of miR-186 in serum exosomes of COPD group was 3.24±0.14,which was significantly higher than that of normal control group 1.03±0.04(t=15.47,P<0.001).The expression level of miR-186 was negatively correlated with the predicted FEV1%of COPD patients(r=-0.5024,p<0.0001).The area under the ROC curve between miR-186 and COPD patients was 0.8409(95%confidence interval:0.6875-0.9943).Conclusion:The high expression of miR-186 in serum exosomes may be a risk factor and may be used as a diagnostic indicator of COPD.

Expression and clinical significance of lncRNA SNHG3 in peripheral blood mononuclear cells of patients with chronic obstructive pulmonary disease

Objective: To investigate the expression of lncRNA SNHG3 in peripheral blood mononuclear cells of patients with chronic obstructive pulmonary disease (COPD), and to explore its clinical significance with chronic obstructive pulmonary disease. Methods: Mononuclear cells were collected from 120 patients with COPD and 110 normal controls. The expression of lncRNA SNHG3 in peripheral blood mononuclear cells was detected by RT-PCR and the difference between COPD group and normal group, and the difference between mild, moderate, severe, extremely severe COPD and health volunteers was analyzed. Results: The relative expression of SNHG3 in peripheral blood mononuclear cells of COPD patients was 1.52 ± 0.52, which was lower than that of the normal control group (2.51 ± 0.59) (P < 0.001). The relative expression of SNHG3 in peripheral blood mononuclear cells of mild COPD was 2.16 ± 0.23, which was higher than that of moderate group (1.55 ± 0.10) (P < 0.001), severe group 1.19 ± 0.11 (P< 0.001), and extremely severe group 0.89 ± 0.06(P < 0.001). In addition, lncRNA SNHG3 can bind to miR-186 and regulate its expression. Conclusions: The expression of lncRNA SNHG3 in peripheral blood mononuclear cells of COPD patients may be a risk factor for COPD and an indicator of the severity of COPD patients. The lncRNA SNHG3/miR-186 regulatory network may play an important role in the development of COPD.

Mir-141 regulates proliferation and apoptosis of lung fibroblasts by targeting ZEB1

Objective:To study the significance of mir-141 expression in COPD(chronic obstructive pulmonary diseases)mononuclear cells,and to demonstrate its effect on proliferation and apoptosis of fibroblasts Possible to ZEB1(zinc finger E-box binding homeobox 1).Methods:40 cases acute phase of COPD patients,80 cases of stable period COPD patients and 110 normal controls in our hospital were collected.RT-PCR was used to detect mir-141 and statistical analysis.The mir-141 mimic was constructed and transfected into lung fibroblasts.The expression of mir-141 was analyzed by RT-PCR.Cell proliferation was analyzed by CCK8 and cell apoptosis was detected by flow cytometry.Targetscan was used to predict the binding site of ZEB1 and mir-141.The target relationship between ZEB1 and mir-141 was identified by RT-PCR and western blot.Results:The expression of mir-141 in mononuclear cells of acute phase of COPD and stable period of COPD was significantly higher than that of normal controls.Mir-141 levels in acute phase of COPD were significantly higher than that of stable period of COPD.Mir-141 mimic transfection significantly up-regulated the expression of mir-141,promoted cell activity and decreased cell apoptosis rate compared with the empty control group.ZEB1 and mir-141 had binding sites predicted by Targetscan database and verified by dual luciferase assays.After mir-141 mimic transfection hat,ZEB1 mRNA and protein expression were down-regulated.Conclusions:High expression of mir-141 in COPD may promote the proliferation of lung fibroblasts and inhibit apoptosis by targeting ZEB1.

Correlation between plasma VEGFA expression level of chronic obstructive pulmonary disease in Li and Han nationalities in Hainan

Objective:To analyze the relationship between plasma vascular endothelial growth factor and patients with chronic obstructive pulmonary disease in Hainan Li and Han nationality, and the correlation between VEGFA and COPD after different stratification, to explore the significance of VEGFA in the diagnosis and treatment of Li people with chronic obstructive pulmonary disease.Methods:249 patients with COPD were recruited in Hainan (109 cases of Li and 140 cases of Han), and 246 cases in the control group (89 cases of Li;157 cases of Han ) From July 2014 to March 2015, We measured plasma vascular endothelial growth factor protein levels ,and determined the expression difference of plasma VEGFA in patients with chronic obstructive pulmonary disease and healthy controls according to different races, genders, smoking status, etc., and analyzed the correlation between the expression of VEGFA in patients with chronic obstructive pulmonary disease in Li and Han in Hainan.Conclusion:(1) In the Li population, the expression of VEGFA of COPD patients and healthy controls was lower than that of Han (P<0.05);(2) The causes of elevated VEGFA levels in the plasma of the Li population are: high BMI, advanced age, use of wood, hay,recent respiratory infections and AECOPD;(3) The causes of elevated VEGFA levels in plasma in Han population are: COPD, low education level, smoking age, repeated respiratory infection, family history of respiratory diseases, frequent coughing, recent respiratory infection and acute COPD Aggravation period;(4) In the AECOPD, the expression of VEGFA in plasma was higher than that in stable phase and control group;the expression of plasma VEGFA in stable phase of COPD was lower than that in the control group.

Correlation between plasma VEGFA expression level of chronic obstructive pulmonary disease in Li and Han nationalities in Hainan

Objective:To analyze the relationship between plasma vascular endothelial growth factor and patients with chronic obstructive pulmonary disease in Hainan Li and Han nationality, and the correlation between VEGFA and COPD after different stratification, to explore the significance of VEGFA in the diagnosis and treatment of Li people with chronic obstructive pulmonary disease.Methods: 249 patients with COPD were recruited in Hainan (109 cases of Li and 140 cases of Han), and 246 cases in the control group (89 cases of Li;157 cases of Han ) From July 2014 to March 2015, We measured plasma vascular endothelial growth factor protein levels ,and determined the expression difference of plasma VEGFA in patients with chronic obstructive pulmonary disease and healthy controls according to different races, genders, smoking status, etc.,and analyzed the correlation between the expression of VEGFA in patients with chronic obstructive pulmonary disease in Li and Han in Hainan.Conclusion:(1) In the Li population, the expression of VEGFA of COPD patients and healthy controls was lower than that of Han (P<0.05);(2) The causes of elevated VEGFA levels in the plasma of the Li population are: high BMI, advanced age, use of wood, hay,recent respiratory infections and AECOPD;(3) The causes of elevated VEGFA levels in plasma in Han population are: COPD, low education level, smoking age, repeated respiratory infection, family history of respiratory diseases, frequent coughing, recent respiratory infection and acute COPD Aggravation period;(4) In the AECOPD, the expression of VEGFA in plasma was higher than that in stable phase and control group;the expression of plasma VEGFA in stable phase of COPD was lower than that in the control group.

Polymerase chain reaction-based assays facilitate the breeding and study of mouse models of Klinefelter syndrome

Klinefelter syndrome(KS)is one of the most frequent genetic abnormalities and the leading genetic cause of nonobstructive azoospermia.The breeding and study of KS mouse models are essential to advancing our knowledge of the underlying pathological mechanism.Karyotyping and fluorescence in situ hybridization are reliable methods for identifying chromosomal contents.However,technical issues associated with these methods can decrease the efficiency of breeding KS mouse models and limit studies that require rapid identification of target mice.To overcome these limitations,we developed three polymerase chain reaction-based assays to measure specific genetic information,including presence or absence of the sex determining region of chromosome Y(Sry),copy number of amelogenin,X-linked(Amelx),and inactive X specific transcripts(Xist)levels.Through a combined analysis of the assay results,we can infer the karyotype of target mice.We confirmed the utility of our assays with the successful generation of KS mouse models.Our assays are rapid,inexpensive,high capacity,easy to perform,and only require small sample amounts.Therefore,they facilitate the breeding and study of KS mouse models and help advance our knowledge of the pathological mechanism underlying KS.