Effects of miR-214 on the proliferation and apoptosis of rheumatoid arthritis synovial fibroblasts by targeting sex determining region Y-box 4 / 中华风湿病学杂志

Objective:To investigate the effect of miR-214 targeting sex determining region Y-box 4 (SOX4) on the proliferation and apoptosis of rheumatoid arthritis (RA) synovial fibroblasts.Methods:Synovial tissue samples from 45 patients with rheumatoid arthritis and 30 patients with joint trauma who underwent joint replacement surgery from 2017 to 2019 in Shangqiu First People's Hospital samples were collected. The expression levels of miR-214 and SOX4 in synovial tissues were detected by real time quantitative polymerase chain reaction (RT-qPCR); The expression levels of serum rheumatoid factor (RF) and C-reactive protein (CRP) were detected by enzyme-linked immunosorbent assay (ELISA) test. The correlation between miR-214 or SOX4 and RF, CRP, erythrocyte sedimentation rate (ESR) was tested by Pearson's analysis. Human fibroblast-like synoviocytes (HFLS)-RA cells were derived from the control group, miR-214 mimic group, mimic-NC group and miR-214 mimic+pcDNA3.1-SOX4 group and se-parated by Lipofectamine 3 000. The cell viability of each group was detected by CCK8, the apoptosis of each group was detected by flow cytometry, the target relationship was verified by dual luciferase experiment, Western-blot was used to detect the protein expression levels of related indicators. Independent sample t test was used for the comparison between the two groups, one-way nalysis of variance (ANOVA) was used for multi-group comparison, and LSD- t test was used for the comparison between the two groups. Results:Compared with the control group (4.6±0.7, 1.9±0.7) and HFLS cells (1.00±0.06, 1.00±0.09), miR-214 was expressed lower in RA tissues (2.6±0.9) and HFLS-RA (0.30±0.05) ( t=10.026, P<0.05; t=15.815, P<0.05). Compared with SOX4 was highly expressed in RA tissues (4.6±0.9) and HFLS-RA cells (3.89±0.41) ( t=14.772, P<0.05; t=12.020, P<0.05). Serum RF, ESR and CRP expression levels in RA group [(46±7) U/ml, (46.4±9.6) mm/1 h, (34.8±9.8) mg/ml] were signifi-cantly higherthan those in the control group [(16±4) U/ml, (9.2±2.0) mm/1 h, (2.1±0.7) mg/ml] ( t=22.906, P<0.05; t=25.338, P<0.05; t=22.314, P<0.05). miR-214 was negatively correlated with serum RF, CRP and ESR was negatively correlated with miRNA( r=-0.574, P<0.05; r=-0.448, P<0.05; r=-0.549, P<0.05), while was positively correlated with SOX4 ( r=0.492, P<0.05; r=0.369, P<0.05; r=0.325, P<0.05). Compared with the control group and the mimic-NC group, cell viability, Ki-67 and p-p65 protein expression decreased significantly after miR-214 mimic transfection for 24 h ( F24 h=16.980, P<0.05;, F48 h=42.735, P<0.05; F72 h=164.448, P<0.05; FKi-67=290.112, P<0.05; Fp-p65/p65=223.548, P<0.05), while the apoptosis rate and Bax protein expression increased significantly ( F=344.360, P<0.05; F=106.376, P<0.05). Overexpression of SOX4, reverses the effect of miR-214. Conclusion:miR-214 targets SOX4 to inhibit the proliferation of synovial fibroblasts and promote apoptosis in rheumatoid arthritis. This may relate to the inhibition of NF-κB signaling pathway by miR-214.

Effects of programmed cell death protein 4 targeting by microRNA-21-5p on the proliferation and apoptosis of peripheral blood B lymphocytes in patients with systemic lupus erythematosus / 中华风湿病学杂志

Objective:To investigate the regulatory effect of miRNA-21-5p on peripheral blood B lymphocyte proliferation and apoptosis in systemic lupus erythematosus (SLE) patients by targeting program-med cell death protein 4 (PDCD4) .Methods:Thirty patients with SLE diagnosed clinically in our hospital were enrolled. Peripheral blood lymphocyte (PBL) was extracted by gradient centrifugation, and B cells were separated by magnetic beads. The proportion of B lymphocyte in peripheral blood was detected by flow cytometry. The cells were divided into five groups by electrotransfection: blank control group, miRNA-21-5p negative control (NC) group, miRNA-21-5p group and miRNA-21-5p inhibitor group, PDCD4 negative control group and PDCD4 siRNA group. Cells were collected 48 hours after transfection. The expression levels of miRNA-21 and PDCD4 were detected by real-time polymerase chain reaction (RT-PCR). Western blotting assay was used to detect the expression of PDCD4 in cells of each group. The targeting relationship between miRNA-21-5p and PDCD4 was verified by double luciferase target experiment. Flow cytometry was used to detect the apoptosis of cells in each group, and CCK-8 method was used to detect the proliferation of cells in each group. Western blotting and RT-PCR were used to detect the expression levels of Fas, FasL, CD40 and CD40L, respectively. Independent sample t test was used to compare the data between the two groups; single factor analysis of variance was used to analyze the results of multiple samples; chi square test was used to compare the positive rate of anti dsDNA antibody. Results:The levels of serum complement [C3 (0.85±0.11) g/L and C4 (0.54±0.09) g/L] in patients with SLE were lower ( t=7.524, P<0.05; t=38.471, P<0.05) than [C3 (1.16±0.17) g/L and C4 (1.57±0.09) g/L] in healthy controls. The levels of anti-dsDNA antibodies (47%), IgG(15.46±0.75) g/L, and IgA (2.68±0.20) g/L were increased than the levels of anti-dsDNA antibodies (17%), IgG (11.95±0.80) g/L, and IgA (2.16±0.11) g/L in healthy controls ( χ2=4.427, P<0.05; t=15.218, P<0.05; t=10.125, P<0.05). The proportion of B lymphocyte [(6.78±0.29)%] and the expression levels of miRNA21-5p (7.52±0.59) in peripheral blood of SLE patients was significantly higher than the proportion of B lymphocyte [(2.03±0.24)%] and the expression levels of miRNA21-5p (3.60±0.62) in healthy controls ( t=59.064, P<0.05; t=19.317, P<0.05), while the expression levels of PDCD4 gene (1.54±0.35) in peripheral blood of SLE patients was significantly lower than that (4.42±0.42) in healthy controls ( t=19.126, P<0.05). Compared with the blank control group and the miRNA-21-5p NC group, cell proliferation in the miRNA-21-5p Inhibitor group was inhibited, and the proportion of apoptotic cells increased ( F=5.244, P<0.05; F=37.903, P<0.05). Compared with the blank control group and PDCD4 NC group, cell proliferation in PDCD4 siRNA group was significantly enhanced, and apoptotic rate decreased ( F=5.956, P<0.05; F=25.431, P<0.05). The results of double luciferase reporter gene assay showed that PDCD4 is the target gene of miRNA-21-5p. Conclusion:miRNA-21-5p may promote the proliferation of peripheral blood B lymphocyte in SLE patients by inhibiting the expression of PDCD4, leading to abnormal lymphocyte apoptosis. miRNA-21-5p can be used as a new target gene for the treatment of systemic lupus erythematosus.

Clinical features and ion channel gene mutations analysis in 17 cases of early-onset epileptic encephalopathy / 中华实用儿科临床杂志

Objective@#To analyze the clinical characteristics and gene mutations of early-onset epileptic encephalopathy(EOEE) caused by ion channel gene mutation, to identify the etiology, to guide the treatment and to provide the basis for genetic counseling.@*Methods@#The clinical data from 17 children with EOEE caused by ion channel gene mutation and the peripheral blood of the children and their parents were collected from June 2014 to May 2018 at the Department of Neurology, Tianjin Children′s Hospital.Epilepsy gene sequencing was performed by using disease gene targeting second generation sequencing technology.The mutation of pathogenic ion channel gene was found.The confirmed mutations were verified by Sanger sequencing and the source of the mutation was identified.@*Results@#Among 17 case with EOEE, 3 cases had genetic mutation, and 14 cases had denovo mutations.Dravet syndrome was found in 8 cases (47.1%), there were SCN1A gene missense mutation in 5 cases, SCN1A gene nonsense mutation in 3 cases, KCNQ2 gene missense mutation in 1 case (5.9%) and non-specific epileptic encephalopathy in 8 cases (47.1%). SCN2A gene missense mutation, SCN4A gene missense mutation, SCN8A gene missense mutation, KCNQ2 gene missense mutation and KCNH gene missense mutation were found in suspected pathogenic mutations.There were 1 missense mutation out of 5 genes, 1 missense mutation of CACNA1A gene, 1 missense mutation of GRIN2A gene and 1 missense mutation of GRIN3A gene.Seventeen patients were treated with 2 or more antiepileptic drugs, 4 with ketogenic diet and 1 with vitamin B6 supplementation.During 11 to 96 months of follow-up, seizures were completely controlled in 3 cases (17.6%), decreased in 7 cases (41.2%) by more than 50%, and decreased in 7 cases (41.2%) by less than 50%.@*Conclusions@#The clinical phenotypes for children with unexplained EOEE are varied, and gene mutations of ion cha-nnel are most common.Some gene sites are denovo mutations which have not been reported such as missense mutation for 3 case SCN1A gene, 1 case SCN2A gene, 1 case CACNA1A gene, 1 case KCNH5 gene, and nonsense mutation for 2 case SCN1A gene, which have enriched the mutation spectrum of EOEE.

Renal protective effect of quercetin on mice with systemic lupus erythematosus and its effect on TGF-β1 and monocyte chemoattractant protein-1 / 中华风湿病学杂志

Objective To study the protective effect of quercetin on the kidney of mice with systemic lupus erythematosus (SLE) and to explore its effect on transforming growth factor (TGF)-β1 and monocyte chemoattractant protein-1 (MCP-1).Methods Thirty BALB/c female mice were randomly divided into control group,model group and drug group according to the envelope method,with 10 mice in each group.A mouse model of SLE was established by intra-peritoneal injection of pristane method.The drug group was given quercetin treatment,and the control group and the model group were given the same dose of normal saline.The renal function index and autoanti-body level in each group of mice were compared.The pathological changes of renal tissues were observed by HE staining.The expressions of TGF-β1 and MCP-1 were determined by Western blotting and real-time fluorescence quantitative reverse transcription polymerase chain reaction (qRT-PCR).The renal function index,autoantibody level,TGF-β1,and MCP-1 expression were statistically analyzed by analysis of one-way analysis of variance (ANOVA).Results The levels of blood urea nitrogen (BUN),serum creatinine (Scr),24 h urine protein,kidney hypertrophy index,antinuclear antibody (ANA) antibody,anti-dsDNA antibody and anti-snRNP/Sm in the model group and the drug group were higher than those in the control group.Compared with the model group,the levels of BUN,Scr,24 h urine protein,kidney hypertrophy index,ANA antibody,anti-dsDNA antibody,anti-snRNP/Sm in the drug group were(11.3±1.1) mmol/L,(45±4) μmol/L,(25.7±2.6) mg/24 h,(4.3±0.4)×10-3,(30.3±3.1) ng/L,(472±48) μmol/L and (17.6±1.8) ng/L,which were decreased (q =10.678,6.698,14.948,14.412,9.226,4.691,8.226,P＜0.01).The glomerular score,tubulointerstitial score and tubulointer-stitial score of the model group were higher than those of the control group.The glomerular score,tubulointer-stitial score and tubular score of the drug group were lower than those of the model group (q=10.935,49.537,20.439,P＜0.01).HE staining showed that the kidney structure of the control group was no obvious damage.In the model group,the glomerular volume of the mice increased,and the inflammatory cells in the renal interstitial and renal tubules infiltrated.The pathological changes in the drug group were significantly reduced compared with the model group.Compared with the control group,the expression levels of TGF-β1,MCP-1 protein and mRNA in the model group and the drug group were significantly increased.Compared with the model group,TGF-β1 and MCP-1 protein and mRNA expression levels the mice in the drug group were significantly reduced.Conclusion Quercetin can improve renal function in mice with SLE by down-regulating the expression of TGF-beta 1 and MCP-1.

Clinical features and ion channel gene mutations analysis in 17 cases of early﹣onset epileptic encephalopathy / 中华实用儿科临床杂志

Objective To analyze the clinical characteristics and gene mutations of early －onset epileptic encephalopathy(EOEE)caused by ion channel gene mutation,to identify the etiology,to guide the treatment and to pro﹣vide the basis for genetic counseling. Methods The clinical data from 17 children with EOEE caused by ion channel gene mutation and the peripheral blood of the children and their parents were collected from June 2014 to May 2018 at the Department of Neurology,Tianjin Children′s Hospital. Epilepsy gene sequencing was performed by using disease gene targeting second generation sequencing technology. The mutation of pathogenic ion channel gene was found. The confirmed mutations were verified by Sanger sequencing and the source of the mutation was identified. Results Among 17 case with EOEE,3 cases had genetic mutation,and 14 cases had denovo mutations. Dravet syndrome was found in 8 cases(47. 1﹪),there were SCN1A gene missense mutation in 5 cases,SCN1A gene nonsense mutation in 3 cases, KCNQ2 gene missense mutation in 1 case(5. 9﹪)and non－specific epileptic encephalopathy in 8 cases(47. 1﹪). SCN2A gene missense mutation,SCN4A gene missense mutation,SCN8A gene missense mutation,KCNQ2 gene missense mutation and KCNH gene missense mutation were found in suspected pathogenic mutations. There were 1 missense mu﹣tation out of 5 genes,1 missense mutation of CACNA1A gene,1 missense mutation of GRIN2A gene and 1 missense mu﹣tation of GRIN3A gene. Seventeen patients were treated with 2 or more antiepileptic drugs,4 with ketogenic diet and 1 with vitamin B6 supplementation. During 11 to 96 months of follow－up,seizures were completely controlled in 3 cases (17. 6﹪),decreased in 7 cases(41. 2﹪)by more than 50﹪,and decreased in 7 cases(41. 2﹪)by less than 50﹪. Conclusions The clinical phenotypes for children with unexplained EOEE are varied,and gene mutations of ion cha﹣nnel are most common. Some gene sites are denovo mutations which have not been reported such as missense mutation for 3 case SCN1A gene,1 case SCN2A gene,1 case CACNA1A gene,1 case KCNH5 gene,and nonsense mutation for 2 case SCN1A gene,which have enriched the mutation spectrum of EOEE.

Establishment of general-special integrated prevention and treatment model for chronic obstructive pulmonary disease patients in community and its preliminary effect / 中华全科医师杂志

A "general-special integrated" prevention and treatment model for chronic obstructive pulmonary disease (COPD) patients in the community was preliminarily established with the joint effets of specialists from the terital hospital and general practitioners in the community health service center.During the implementation of the model the general practitioners recieved research training and participanted in the research project of COPD management;and the "general-special integrated" outpatient clinic greatly improved the management for COPD patients.Since the establishment of the model,the number of acute attacks of COPD patients was decreased,and the proportion of standardized medication was increased.The model also improved the research ability and clinical competency of general practitioners.The established model provides experiences for the tiered-management for COPD patients in the community.

Development of Intelligent Management System for Diabetes Medication Knowledge Base / 中国药师

Objective:To develop the intelligent management system for diabetes medication knowledge base with the help of community hospital information platform.Methods:Based on the China Type 2 Diabetes Prevention Guide (2013 edition),National Essential Drugs (2012 edition) and the relevant package inserts,the knowledge base of diabetes medication was established.Results:The information platform could make interception beforehand,warning in the process of medication and provide attentions for physicians after treatment to determine rational medication.Conclusion:The intelligent management system for diabetes medication knowledge base can promote physicians to firmly grasp the principles of drug treatment,simplify treatment regimen,select effective drugs with reasonable prices and provide individualized treatment.

Establishment of the management model of chronic obstructive pulmonary disease patients in the Gumei community based on internet of things / 复旦学报（医学版）

Objective On the basis of internet of things technology,to initially establish a management model of chronic obstructive pulmonary disease (COPD) patients in Gumei community,so as to provide experience for the comprehensive management of community COPD patients.Methods According to the characteristics of the Internet of things technology,we formulated a scheme as a technical route to manage the COPD patients.A homogenous COPD management team of doctors was established under the training of experts from the Department of Respiration of Zhongshan Hospital,Fudan University.Results We drew a COPD patient management model chart,and initially formed a qualified and homogeneous COPD management team of general practitioner.Conclusions Through the Internet of things technology management,we initially formed a set of manual quality control model in the process of data automatic transmission,and initially formed a management model of community COPD patients,based on the internet of things.