Objective To observe the electrocardiogram (ECG) of urban residents in non-endemic areas of Keshan disease in Ji'nan City,Shandong Province,and to provide scientific basis for health evaluation and prevention and control of Keshan disease.Methods From July 2014 to June 2015,ECGs of residents undertaken healthy examination in Shandong Ciming Physical Examination Center were collected,retrospective analysis was used to analyze residents' heart health.Statistical analysis was conducted according to sex and age groups,and the diagnosis of ECG was based on Concise Pediatric Electrocardiogram and Clinical Electrocardiography.Results Totally 31 753 ECGs were recorded,including 18 664 males accounting for 58.8％ and 13 089 females accounting for 41.2％,who were aged 6-78 years old,including 304 people 6-20 years old,18 861 people 21-40 years old,10 451 people 41-60 years old,and 2 137 people 61-78 years old.Totally 27 699 normal ECG (87.2％) and 4 054 abnormal ECG (12.8％) were detected,there were 2 212 males and 1 842 females in the abnormal ECG,the detection rates were 11.9％ (2 212/18 664) and 14.1％ (1 842/13 089) for males and females,respectively.The detection rate of female was significantly higher than that of male (x2 =34.082,P ＜ 0.01).The detection rate of ST-T change in abnormal ECG was the highest,which was 316.5/10 000 (1 005/31 753),men and women were 273.3/10 000 (510/18 664) and 378.2/10 000 (495/13 089),respectively.The detection rate of arrhythmia was 4.8％ (1 523/31 753),including 1 023 males and 500 females,and their detection rate of arrhythmia was 548.1/10 000 (1 023/18 664) and 382.0/10 000 (500/13 089),respectively.In arrhythmia,degree Ⅰ atrioventricular block (155.3/10 000,493/31 753),complete rightbundle branch block (81.6/10 000,259/31 753),ventricular premature beats (79.0/10 000,251/31 753),and atrial premature beats (48.2/10 000,153/31 753) had higher detection rates.Conclusion The detection rate of women of urban residents in Ji'nan is higher than that of men,the most common abnormal ECG is ST-T change,and the detection rate of arrhythmia is low.

Objective To provide more valuable information for diagnosis of dilated cardiomyopathy (DCM),we detected differentially expressed proteins in serum from patients with DCM and healthy people and protein biomarkers were selected.Methods During the period from march 2011 to may,a total of 29 samples of resident patients with DCM from Qilu Hospital of Shandong University,Jinan Central Hospital and Jinan First Peoples Hospital and 30 local healthy people in Jinan were selected as DCM and control groups,respectively.Serum samples from these patients with DCM and controls were detected by ClinProt MALDII-TOF-MS.ClinProTools 2.2 software was used to get mass spectrometric data.The ClinPrott discrimination model was established to screen out differentially expressed proteins as potential biomarkers.Results Via comparing proteins/polypeptides peaks of DCM patients and healthy controls,57 of all 73 peaks were found to be significantly different between the two groups.Compared with the control group,35 peaks were up-expressed while the other 22 peaks were downexpressed.Five peaks were screened out as protein biomarkers.They were mass-to-charge ratio (m/z):4 247.95,4 209.37,1 058.69,1 074.78,and 2 364.71.The sensitivity and specificity of ClinPrott discrimination model was 99.14％ and 99.16％,respectively.Conclusion Patients with DCM have expressed serum proteins differently and we have found five protein markers which might have some value for diagnosis of DCM.

Objective To do screening acute lymphoblastic leukemia patients scFv antibody single chain variable region to cre-ate conditions for the expression and obtain further specificity of antibody fragments.Methods In this study,patients with newly diagnosed acute lymphoblastic leukemia serum as coating antigen using phage display technology,screening phage an-tibody specificity from the semi-synthetic human phage antibody libraries,the first to target the immune antigen-coated tab-let,phage library was added,so that with the target antigen-specific binding phage antibody was immobilized on plates immu-nization,could not be specifically bound phages were rinsed.The eluted specific binding phage,E.coli infection.Could get the specific antibody gene containing phagemid.Results After three “adsorption-elution-amplification”screening process,got stronger leukemia patient antigen-specific phage antibody variable region fragment and identification.Conclusion Got better strain affinity antibody fragments,to create the conditions for the next fragment expression,identification and clinical appli-cation.

Objective To investigate the clinical diagnostic value and pathogenesis of serum protein identification in Keshan disease (KD).Methods A total of 65 chronic KD patients were selected as the patient group in KD endemic areas,while 29 cases of dilated cardiomyopathy (the DCM group),62 healthy cases from KD endemic areas (control 1 group) and 28 healthy cases from non-endemic areas (control 2 group) were selected as controls.Liquid chip time of flight mass spectrometry (ClinProtTM MALDI-TOF-MS) was used to determine the expression of proteins/peptide peaks.ClinProTools 2.2 software was used to analyze the protein profiles to determine differentially expressed proteins/peptide peaks.The Genetic Algorithm (GA),QuickClassifer Algorithm (QC) and Supervised Neural Network Algorithm (SNN) methods were used to screen marker proteins.Matrix-assisted laser desorption/ionization time-of-flight Mass Spectrometry technique (MALDI-TOF/TOF) was also used as a secondary mass spectrometry to identify differentially expressed peptides.Results Between the KD and control 1 groups,34 differentially expressed proteins/peptides and 5 marker proteins were identified,while 52 differentially expressed proteins/peptides and 5 marker proteins were identified between the KD and control 2 groups,and there were 67 differentially expressed proteins/peptides and 5 marker proteins between the KD and DCM groups.During secondary mass spectrometry,two peptides for mass-to-charge ratio (m/z) 2 079 and 1 465 were obtained,peptide of matching β-globin showed low expression while peptide of matching fibrinogen showed high expression in the KD patients.Conclusions Serum marker proteins can be used as biomarkers for diagnosis and differentiation of KD.β-globin and fibrinogen play an important role in the development of KD myocardial injury.

Objective To screen permethrin human single-chain variable region (scFv) antibody for aims of developing rapid detection kit. Methods Phage display technology was used in this study. Permethrin was solid phase coated on Nunc plate as antigen. Semi-synthetic single-chain variable region of human antibody library technology was applied, and single chain variable region was screened from phage antibody library after 3 rounds "adsorption - elution - amplification" of the selection process. 100 clones were random selected as resistance to permethrin clones , enzyme-linked immunosorbent assay (ELISA), crossreactivity and competitive inhibition experiments were used to validate permethrin binding activity with strong scFv clones from the selected phage antibody clones plasmid. The plasmid was digested with restriction enzyme Sfi Ⅰ / Not Ⅰ and subcloned into pCANTAB5E vector. After transformed into E. coli XL1BIue, the plasmid was identified by restriction enzyme analysis. Results After screening in 100 clones, 18 clones had high ELISA absorbance values ( A value) at 490nm wavelength ( A490nm), then bovine serum albumin (BSA) cross-reactions identified five weak cross-reaction. Combined with the triplicate ELISA and competitive inhibition experiment results, one positive clone was acquired at last. And this clone was subcloned into pCANTAB5E vector and transformed into competent cells XL1-Blue. Conclusion Plasmid fragment was consistent with the purpose, which provided the foundation for further study of its specific affinity.

Seventeen kinds of serum free amino acids (FAA) of 35 healthy persons in endemic area of Keshan disease and 38 patients with Keshan disease were determined and compared with 31 healthy persons in non-endemic area. It was found that mean values of total FAA, essential amino acids, non-essential amino acids and serum protein of inhabitants in endemic area of Keshan disease were significantly lower(P

The problem of Keshan disease (KD) is confused with dilated cardiomyopathy (DCM) is still not solved.KD and DCM have different gene expression profiles,namely,different Micro RNAs (miRNAs) and Long noncoding RNAs (lncRNAs).There may be characteristic miRNAs and lncRNAs in KD and DCM.Systemically study differential gene expression profiles and regulation of noncoding RNAs gene expression and elucidate the different molecular pathogenesis in gene expression and gene expression regulation will provide theoretic basis in identification,prevention and treatment of KD and DCM.

Objective To investigate the differences in gene expression profiles of peripheral blood from patients with Keshan disease (KD) and the apoptosis mechanism in KD,to obtain diagnostic markers and establish diagnostic centroids plot for KD.Methods RNA was isolated from ten patients with KD diagnosed according to the clinical criteria for KD in China and ten health controls.The expression profiles were evaluated by Agilent 4 ×44K Whole Human Genome density oligonucleotide microarray analysis.The data were extracted by Agilent Feature Extraction Software t test,Pathway studio analysis and prediction analysis for microarray (PAM) were used to identify differently expressed genes,gene pathways,diagnostic markers and establish diagnostic centroids plot.Results Totally 1 570 up-regulated genes and 1 498 down-regulated genes were identified.Thirty-eight enrichment pathways were also identified,and the highest ranked by Pathway studio analysis was related to apoptosis.Six genes involved in apoptosis pathway were up-regulated in KD included ataxia telangiectasia mutated (ATM),cAMP-dependent protein kinase,protein kinase A (PKA),baculoviral IAP repeat-containing 2 (BIRC2),NLR family,apoptosis inhibitory protein (NAIP),BCL2-1ike 11 (Bim),BCL2-related protein A1 (BCL2A1) and down-regulated were 7 which included caspase 8 (CASP8),BCL2 binding component 3 (BBC3),BCL2--associated athanogene (BAG1),BCL2-associated X protein (BAX),BCL2-1ike 1 (BCL2L1),BCL2-related ovarian killer (BOK),and caspase 6 (CASP6).Forty-two diagnostic markers were obtained through PAM analysis.Conclusions Apoptosis related to genes and pathways might play an important role in the pathogenesis of KD.Forty-two markers could be used as molecular markers for the diagnosis of KD,which is important to the diagnosis of KD.

Objective By constructing the differential expression profile of lncRNA/mRNA in peripheral blood plasma of patients with Keshan disease (KSD) and dilated cardiomyopathy (DCM),to explore the commonality and characteristics of the two diseases in molecular mechanism.Methods Ten patients with chronic KSD were selected in the severe disease area of KSD in Shandong Province,and 10 cases of DCM and 10 healthy subjects (control group) were selected in non-KSD area.Blood of elbow vein was collected and plasma was separated.RNA-seq technology was used to construct the differential lncRNA/mRNA expression profile between KSD and control group,DCM and control group,and co-expression and specific expression of partial genes in KSD and DCM were analyzed through Wien analysis.The lncRNA-mRNA co-expression network maps of specific part of KSD,specific part of DCM and common part of the two diseases were constructed,and Gene Ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were applied to distinguish the biological function of the two diseases.Results Compared with control group,102 dysregulated mRNAs and 22 dysregulated lncRNAs showed the same trend in KSD and DCM.And 3 606 mRNAs and 451 lncRNAs were only differentially expressed in KSD group,217 mRNAs and 137 lncRNAs were only differentially expressed in DCM group.The differentially expressed lncRNA/mRNA shared between the KSD and DCM groups were mainly about viral transcription,immuno-inflammatory response,oxidative stress signaling pathways.The KSD specific lncRNA/mRNA mainly participated in cell membrane damage and viral myocarditis.The DCM specific lncRNA/mRNA mainly regulated mitochondrial structure and oxidative phosphorylation related enzymes.Conclusion The differentially expressed lncRNA/mRNA shared in KSD and DCM groups are mainly involved in viral transcription,oxidative stress signaling pathways;KSD specific lncRNA/mRNA are mainly related to cell membrane damage and viral myocarditis;DCM specific lncRNA/mRNA mainly regulate mitochondrial structure.

Objective:Through differential miRNA expression profiles and bioinformatics in the peripheral blood of patients with Keshan disease (KD) and healthy control, to explore the possible pathogenesis of KD.Methods:Ten patients with chronic KD (KD group) were selected in the severe disease area of KD in Wulian County, and 10 healthy subjects (control group) were selected in non-KD area of Dongchangfu District, Shandong Province. Blood sample of elbow vein was collected and plasma was separated. RNA-seq technology was used to construct the differential expression profiles of miRNA in KD and control groups. Target mRNAs were screened using Starbase, miRTarBase, miRDB and TargetScan. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were conducted to investigate the possible pathogenesis of KD.Results:Compared the control group and KD group, 132 differentially expressed miRNAs were screened out, including 90 upregulated and 42 downregulated miRNAs. Through Starbase, miRTarBase, miRDB and TargetScan, 53 miRNAs were obtained, 737 targeted mRNAs were obtained. GO analysis showed that the differential genes were mainly involved in the biological processes of Ras protein signal transduction, transmembrane transport, cell cycle regulation, cell adhesion, etc. KEGG pathway analysis showed that the differential genes were mainly involved in viral infection, endocytosis, adhesion spot and actin regulation.Conclusion:In this study, RNA-seq technology is used to obtain differential miRNA expression profiles of KD patients and healthy control, and target pathogenic genes and signaling pathways that may be related to KD are screened out.