Objective:To investigate the effect of lipopolysaccharide (LPS) on primary neonatal rat cardiomyocytes (CMs) and human induced pluripotent stem cell-derived cardiomyocytes (hiPS-CMs).Methods:The hiPS-CMs and primary neonatal rat CMs were treated with different concentrations of LPS for 24 to 48 h. Then the cellular viability was analyzed by the xCELLigence RTCA Cardio system. The measurement of NPPB gene was studied by qRT-PCR and the gene expression analysis was performed by the qPCR array, in order to evaluate the cardiac inflammation effect induced by LPS.Results:The LPS exposure led to dysfunction in the primary neonatal rat CMs, which shown as an increase in beating rate and a decrease in contraction amplitude ( P<0.01), accompanied by an increased NPPB mRNA level ( P<0.01). There was no significant alteration in beating rate and the contraction amplitude in the corresponding concentration of the primary neonatal rat CMs ( P>0.05), as well as the NPPB mRNA level ( P>0.05). However, the expression of NPPB mRNA in hiPS-CMs was significantly different at a higher concentration of LPS (5 μg/mL~40 μg/mL) ( P<0.01), but the beating rate and the contraction amplitude showed no significant change, even the concentration of LPS up to 40 μg/mL ( P>0.05). Finally, the genes of C3, Gpnmb, Atf3, Il6r and Ly96 upregulated to 1.5 folds in the primary neonatal rat CMs. In comparison with primary neonatal rat CMs, the AK4, TOLLIP, SPP1, FABP1, IL6R, LY96 and C3 were over expression to 1.5 folds in the hiPS-CMs. Conclusions:In comparison with primary neonatal rat CMs, hiPS-CMs are markedly less injured by LPS and show a different pattern of inflammation gene expression.

Objective:To analyze the sepsis related long non-coding RNA (lncRNA) and mRNA expression profiles based on Gene Expression Omnibus (GEO) datasets and bioinformatic analysis, and to analyze the sepsis-associated competing endogenous RNA (ceRNA) network based on microRNA (miRNA) database.Methods:The sepsis-related lncRNA dataset was downloaded from the GEO database, and the differential expression analysis was conducted by Bioconductor on the sepsis dataset to obtain differentially expressed lncRNA (DElncRNA) and differentially expressed mRNA (DEmRNA), and cluster heat map was drawn. miRNA combined with DElncRNA were predicted by miRcode. mRNA targeted by miRNA was simultaneously met by three databases: TargetScan, miRDB, and mirTarBase. The interaction relationship of lncRNA-miRNA-mRNA was obtained. The regulatory network visualization software CytoScape was used to draw ceRNA networks. DEmRNA in the ceRNA networks were imported into the Search Tool for the Retrieval of Interacting Genes Database (STRING) online database to draw the protein-protein interaction (PPI) map. The gene ontology (GO) function annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis of DEmRNA were performed.Results:Dataset GSE89376 and GSE145227 were found from GEO database. Difference analysis showed there were 14 DElncRNA and 359 DEmRNA in the elderly group of GSE89376; 8 DElncRNA and 153 DEmRNA in the adult group of GSE89376; 1 232 DElncRNA and 1 224 DEmRNA in the children group of GSE145227. Clustering heatmap showed that there were significant differences in the expression of lncRNA and mRNA between the sepsis group and the control group. The ceRNA networks were constructed with miRNA. Several DElncRNA and multiple DEmRNA participated in the ceRNA network of sepsis. The PPI diagram demonstrated that several genes encoding proteins interacted with each other and form a multi-node interaction network with multiple genes encoding proteins. Functional annotation and enrichment analysis demonstrated that there might be a crosstalk mechanism on functionally related genes such as nuclear receptor activity, ligand-activated transcription factor activity, and steroid hormone receptor activity, and played a role in the occurrence and development of diseases through forkhead box transcription factor O (FoxO) signaling pathway, Janus kinase/signal transducers and activators of transcription (JAK/STAT) signaling pathway, p53 signaling pathway, and phosphateidylinositol 3-kinase (PI3K)/Akt signaling pathway.Conclusion:Through sepsis-related lncRNA-miRNA-mRNA ceRNA network and combining with KEGG pathway analysis, there were several lncRNA and mRNA participating in the ceRNA network related sepsis, which played an important role in several signal pathways.

This experimental study was designed to explore the possible mechanisms of Cariporide, a kind of Na+/H+ exchanger inhibitor, for protecting the lung from warm ischemia/reperfusion injury (WI/RI) of isolated rat lung model. Thirty isolated rat lungs were established on the Langendorff apparatus and randomly divided to three groups (n = 10, each): control group (C group), ischemia/reperfusion group (IR group) and Cariporide group (CP group). Mean pulmonary artery pressure (MPAP) and peak airway pressure (pAwP) were monitored continuously. At the end of reperfusion, right bronchoalveolar lavage was performed, bronchoalveolar lavage fluid (BALF) recovery rate (BALFRR) was recorded, and protein content in BALF was measured. Lung water content (LWC), malondialdehyde (MDA) and superoxide dismutase (SOD)of left lung tissue were measured; histomorphology evaluation was performed under light microscope and transmission electron microscope. In comparison with the data from IR group, BALF protein concentration, LWC, MDA content and MPAP content of reperfusion were significantly decreased, but SOD activity was increased in CP group. Histomorphologic feature also showed that pathological change significantly reduced in CP group. In this rat WI/RI model, the mechanism by which the selective Na+/H+ exchanger inhibitor (Cariporide) attenuates lipid peroxidation induced by WI/IR may be: preventing Ca2+ overload via inhibiting the transport of Na+/H2 exchanger-1 (NHE1) in the context of the coupled exchanger, thereby reducing the activation of xanthine oxidase pathway and oxygen free radical liberation which is dependent on certain intracellular Ca2+ concentration, and lastly promoting the endogenous antioxidative mechanism.
Animals ; Antioxidants ; pharmacology ; Guanidines ; pharmacology ; In Vitro Techniques ; Ischemic Preconditioning ; Lung ; blood supply ; Male ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; etiology ; prevention & control ; Sodium-Hydrogen Exchangers ; antagonists & inhibitors ; Sulfones ; pharmacology ; Superoxide Dismutase ; metabolism ; Warm Ischemia

Objective To investigate the effect of 5-AZA-2'-dC on Angiotensin Ⅱ (Ang Ⅱ)-induced cardiomyocyte hypertrophy.Methods Cultured cells derived from neonatal heart of rat were divided into 5 groups:normal control,hypertrophic group,5-AZA-2'-dC treatment group,and 5-AZA-2'-dC pretreatment group.Neonatal rat cardiomyocyte hypertrophic response was assayed by the size of cardiomyocytes and atrial natriuretic polypeptide (ANP) expressive level.The level of sarcoplasmic reticulum Ca2+ ATPase (SERCA2a),total calmodulin kinase Ⅱ (CaMK Ⅱ) and phospho-CaMK Ⅱ (p-CaMK Ⅱ) detected by Western blot.The intracellular calcium changes of cardiomyocytes were imaged by confocal fluorescent microscopy.Results Cells treated with Ang Ⅱ at 10-6 mol/L for 48 h were chosen as hypertrophic cardiomyocyte model.The mRNA expression and protein level of ANP were significantly decreased in the treatment and pretreatment groups compared with hypertrophic group.The protein level of SERCA2a was significantly decreased in the hypertrophic group,and increased in the treatment and pretreatment group compared with hypertrophic group.The protein level of SERCA2a was significantly decreased in the hypertrophic group,and increased in the treatment and pretreatment group compared with hypertrophic group,whereas phospho-CaMK Ⅱ showed an opposite change tendency.The time required for increasing and declining to half of the intracellular calcium peak value were both delayed in hypertrophic group,as the treatment and pretreatment groups showed shorter time required compared with hypertrophic group.Conclusion 5-AZA-2'-dC could inhibit Ang Ⅱ-induced cardiomyocyte hypertrophy which might be related to regulate SERCA2a expression.Increased SERCA2a expression may maintain the calcium homeostasis through shortening the time of transfer Ca2+ from the cytosol of the cell to the lumen of the sarcoplasmic reticulum.

Objective: This study aimed to investigate the correlation between supraventricular tachycardia (SVT) and tachycardia-induced cardiomyopathy(TIC) in children and assess the risk factors for the development of TIC. Method: One hundred and eighty-three patients were recruited in four hospitals of Beijing diagnosed as SVT were divided into two groups depending on whether or not complicated with TIC. The age, gender, tachycardia type, ventricular rate during tachycardia, frequency of tachycardia episodes and duration of tachycardia were compared between the two groups and risk factors associated with TIC were analyzed. Statistical analyses were performed using t test, Mann-Whitney U and χ² test. Result: Totally 183 patients were included in this study (paroxysmal supraventricular tachycardia, PSVT, n=149; atrial tachycardia, AT, n=34). The incidence of TIC was 13.1%. There was significant difference regarding the incidence of TIC between children with AT and children with PSVT (26.5% vs. 10.1%, χ²=6.537, P=0.011). The percentage of AT in patients with TIC was significantly higher than patients without TIC (37.5% vs. 15.7%, χ²=6.537, P=0.011). Patients with TIC had a higher frequency of tachycardia episodes(2.5(1.0-4.8) beats/year vs 4.0(1.0-10.0) beats/year, Z=-2.223, P=0.026)and longer duration of tachycardia(11.0(1.1-36.0) h vs 2.5(1.0-12.0) h, Z=-2.154, P=0.031)compared with patients without TIC. Multivariate predictors of TIC was longer duration of tachycardia (OR=2.041, P=0.028). Conclusion TIC occurs in 13.1% of children with SVT. The incidence of TIC in children with AT is higher than in children with PSVT. Risk factor for the development of TIC is longer duration of tachycardia.

Objective: The study assessed the clinical characteristics and response to acute intravenous antiarrhythmic drug therapy of supraventricular tachycardia (SVT) in children. Methods: This was a multicenter prospective descriptive study including 257 children from First Hospital of Tsinghua University, Peking University First Hospital, Children's Hospital Affiliated to Capital Institute of Pediatrics and Beijing Anzhen Hospital who received intravenous antiarrhythmic drug therapy for SVT from July 2014 to February 2017. The clinical and tachycardia features, response to intravenous antiarrhythmic drug therapy of these children were characterized. Statistical analyses were performed using t test, Mann-Whitney U test, χ² test and H test. Results: The onset of SVT occurred at any age with a distribution with positive skewness, 57.6% (n=148) children<1 year, 17.5% (n=45) children1~<3 years, 10.5% (n=27) children 3~<6 years and 14.4% (n=37) children ≥ 6 years of age. The percentages of SVT types were 49.4% (n=127) for atrioventricular reentry tachycardia (AVRT), 4.3% (n=11) for atrioventricular nodal reentry tachycardia (AVNRT), 26.8% (n=69) for unclassified paroxysmal SVT and 19.5% (n=50) for atrial tachycardia (AT), respectively. Tachycardia-induced cardionyopathy (TIC) secondary to SVT developed in 30 of 225 (13.3%). Left ventricular ejection fraction (LVEF) of the 27 children attacked by TIC returned to normal after successful control of SVT (41.1%±6.3% vs. 60.3%±9.2%, t=-10.397, P=0.000). Complete termination of SVT by antiarrhythmic drugs was achieved in 164 of 257 (63.8%), partial termination rate was 18.7% (48 of 257) and failure to terminate rate was 17.5% (45 of 257). Propafenone (complete cardioversion in 98 (73.1%) of 134) and amiodarone (complete cardioversion in 23 (76.7%) of 30) showed better efficacy for SVT termination than adenosine (complete cardioversion in 26 (44.1%) 59) (χ²=20.524, P=0.000). Paroxysmal SVT had a higher termination rate on pharmacological therapy than AT (67.1% vs. 50.0%, χ²=6.337, P=0.042). Patients of different age groups had significantly different response to antiarrhythmic therapy (χ²=13.904, P=0.031). Children<1 year of age showed the least response to antiarrhythmic drug therapy with complete termination in 51 (55.4%) of 92. Adverse effects occurred in 9 patients (3.5%): Four patients had severe hypotensive shock using propafenone (n=3) and adenosine (n=1), and 3 patients had sinus arrest using adenosine. Conclusion Most (57.6%) children with SVT have their first clinical episode within 1 year of age, and AVRT is the most common type. TIC occurs in 13.3% of children with SVT. Intravenous antiarrhythmic drug therapy has a 63.8% complete termination rate for children with SVT and incidence of adverse effects is 3.5%. Propafenone and amiodarone are more effective for SVT termination in children than adenosine. Serious adverse effects may occur when using propafenone.

The aim of this study was to investigate the correlation between porcine epidemic diar-rhea virus(PEDV)S1 IgG antibody levels and neutralizing antibody potency in sow sera.Sera from 5 PEDV-infected farms with a clear immune background,5 non-infected farms and 5 infected farms with an unclear immune background,and sera from return-fed reserve pigs,totaling 716 copies,were collected and measured,and the correlation between PEDV S1 IgG antibodies and neutralizing antibodies was analyzed.The results showed that the PEDV S1 IgG and neutralizing antibodies of sow sera showed highly significant positive correlation,the correlation coefficient was 0.892(P＜0.000 1).Previous studies have shown that the level of PEDV neutralizing antibodies in sow serum correlates with the ability of piglets'maternal antibodies to resist PEDV infection.Therefore,the a-bility of maternal antibodies against PEDV in piglets can be evaluated by detecting PEDV S1 IgG antibodies in the serum of sows.In 10 PEDV-infected farms,the neutralizing antibodies to PEDV in the sera of sows after immunization were generally high,and the S1 IgG antibodies were also high,and their S/P values were higher than 3.5 in 66.9％of the farms(347/519),and the highest anti-body levels were found in the four farms in which PED did not occur,whereas the neutralizing an-tibodies in the immunized sows in the five PEDV-uninfected farms were generally low,and their S1 IgG antibodies were also low,and only 8.1％(13/161)having S/P values higher than 3.5.The re-sults suggest that most sows in PEDV-infected farms can provide good immunoprotection to pig-lets after immunization,while pigs in PEDV-uninfected farms need further immunization if they need to achieve a higher level of immunoprotection.The present study provides a substantial clini-cal basis for the use of PEDV S1 IgG antibody levels to assess the effectiveness of PEDV antibody protection in swine herds.

The aim of this study is to establish a simple and accurate method for vaccine immune e-valuation of porcine pseudorabies virus.In this research,PRV-gD recombinant protein was ex-pressed from mammalian cell HEK-293F as coating antigen,and then the reaction conditions of gD-iELISA were optimized according to checkerboard titration method.The gD-iELISA was used to detect the antibody levels of 211 clinical pig serum samples and the consistency with the neu-tralizing antibody levels wasanalyzed.The results showed that the antigen coating concentration was 0.90 mg/L;the serum to be detected was diluted 1∶100 and incubated at 37 ℃ for 30 min;goat anti-pig IgG-HRP antibody was diluted 1∶55 000 and incubated at 37 ℃ for 30 min;TMB sub-strate was developed at 37 ℃ for 20 min.The method could detect 1∶6 400 diluted PRV positive serum.The results of CSFV,PRRSV,PCV-2,PEDV and FMDV positive sera were all negative by gD-iELISA,and there was no cross-reaction between the method and the above positive sera.The coincidence rate of gD-iELISA and commercial kits was 95.26％,and the intra-and inter-batch co-efficients of variation were both less than 10％.Correlation analysis showed that the correlation coefficient(r)between gD antibody level and neutralizing antibody level was significantly greater than that of gB antibody level,and the gD antibody level had a good linear relationship with the neutralizing antibody level.The results indicated that gD-iELISA was more suitable for vaccine im-mune evaluation of PRV than gB-iELISA.Therefore,the method will have a good prospect of ap-plication in the immunization control of the PRV.