Objective Explore the relationship of ventricular late potential (VLP) and the heart function of children with viral myocarditis,and provide the evidence for their diagnosis and therapy.Methods The clinical data of 152 cases of epileptic children were collected.The patient group was divided into three groups (severe arrhythmia,heart failure,and cardiogenic shock).The patient group was also divided into two groups (cardiac dilatation,and non-cardiac dilatation) according to UCG.Serum levels of cTnI and VLP in children with viral myocarditis were detected.Results The VLP was negative in the mild and control groups,but the positive rate of VLP is 75.0 ％ in the severe group.The positive rate of VLP was 60.0％ in the severe arrhythmia group,77.78％ in the heart failure group,and 100％ in the shock group.There is one kind of negative rank correlation between LAS,RMS40 and LVFS (P ＜ 0.05),and another negative rank correlation between RMS40 and LVEF (P ＜ 0.05).Conclusions The children with viral myocarditis have a favorable prognosis.The sever patient in the minority must be diagnosed in time and treated because of the critical state of viral myocarditis children.The serum level of cTnI and VLP were increased in children with viral myocarditis,and they were sensitive parameters to reflect patients’ condition.

BACKGROUND: The implantation technique of bone marrow mesenchymal stem cells (MSCs) is of important significance for repair of brain injury. However, its action pathway still needs to be investigated.OBJECTIVE: To observe the changes of Bcl-2 and Bax expressions in the injured regions of cerebral cortex and hippocampus of rats with cerebral ischemia in which MSCs were implanted, and to analyze the action mechanism of intracranial implantation of MSCs inhibiting neuronal apoptosis.DESIGN: A randomized controlled experiment.SETTING: Institute for Cerebrovascular Disease, the Affiliated Hospital of Qingdao University Medical College.MATERIALS: Twenty-four male adult Wistar rats, weighing 180 to 240 g, were provided by the Experimental Animal Center of Shandong University. The involved rats were randomized into 3 groups with 8 in each: control group, injury group and implantation group. 5-bromodeoxyuridine (BrdU, Sigma Company), TUNEL kit, Bcl-2, Bax antibody kit were purchased from Beijing Zhongshan Bioengineering Company.METHODS: This study was carried out in the Institute for Cerebrovascular Disease, the Affiliated Hospital of Qingdao University Medical College between April 2005 and September 2006. Rats in the injury group and implantation group were developed into rat models of cerebral ischemia by suture of external carotid artery. Seven days later, the successful rat models in the implantation group were injected in the cerebral cortex and striatum with 2×1012 L-1 MSCs suspension primarily cultured in vitro. The processing of the experimental animals corresponded to the requests of Animal Ethics.MAIN OUTCOME MEASURES: Neuronal apoptosis and Bcl-2 and Bax expressions in the injured regions of cerebral cortex and hippocampus were detected by TUNEL staining and immunohistochemical method on the 7 and 14 days after successful modeling, separately.RESULTS: ①Neuronal apoptosis: On the 7th day after successful modeling, apoptotic cells were not found in the control group, and apoptotic cells in the implantation group were significantly fewer than those in the injury group (P ＜0.01). ② Bcl-2 and Bax expressions: On the 14th day after successful modeling, Bcl-2-positive neuronal expression in the cerebral cortex and hippocampus of rats in the injury group was significantly weaker than that in the control group and implantation group (P ＜ 0.01). Bax-positive expression in the cerebral cortex and hippocampus of rats in the injury group was significantly stronger than that in the control group and implantation group (P ＜ 0.01).CONCLUSION: MSCs can promote Bcl-2 expression and inhibit Bax expression of rats with cerebral ischemia injury,and accordingly neuronal apoptosis will be reduced.

Objective To detect and determine the specificity of platelet-reactive antibodies in patients who were refractory to platelet transfusions.Methods Serum samples from 48 patients who were refractory to platelet transfusions were screened with MACE for platelet-reactive antibodies.Specificity of platelet alloantibodies was determined with PAK12 and MAIPA.Results Platelet-reactive antibodies were detected in the serum of 50% PTR patients(24/48).The incidence of HLA antibodies was 39.6%(19/48),accounting for 79.2% of serum with platelete alloantibodies.The HPA alloantibodies were found in 29.2%(14/48)serum,of which,64.3%(9/14)occurred together with anti-HLA.The following platelet-specific antibodies were identified:anti-HPA-3a(n=2),anti-HPA-5b(n=1),anti-HPA-5a(n=1),anti-HPA-2b(n=1),anti-HPA-4b(n=1).Of the 14 serum with HPA antibodies,78.6%(11/14)contained panreactive anibodies against platelet glycoprotein(GP)Ⅱb/Ⅲa,GPⅠa/Ⅱa,and/or GPⅠb/Ⅸ.Platelet-reactive antibodies were detected more in female(16/29)than in male(8/19)with a frequency of 55.2%,42.1%,respectively,but there was no statistical significant difference.Conclusion The platelet-specific antibody in PTR patients are not as rare as previous thought although alloantibodies are predominantly anti-HLA.Antibody specificities in Chinese PTR patients are different from those observed in Caucasians,in whom anti-HPA-5b and-1b are the most prevalent specificity.The most prevalent platelet-specific antibodies are anti-HPA-3 and anti-HPA-5 while anti-HPA-4b and anti-HPA-2b are also detected.

AIM:To research the characteristics of ventricular electrophysiology in right ventricular rapid pacing-induced congestive heart failure (CHF) dogs. METHODS:Dogs (n = 16) were randomly divided into 2 groups:the control (n = 7) and the CHF group (n = 9) induced by rapid right ventricular pacing at 240 pulse·min-1 for 4 to 5 weeks. The electrophysiologic parameters were evaluated by the technique of standard electric stimulation and monophasic action potential (MAP) recording. RESULTS:(1) Ventricular effective refractory period (VERP), ventricular MAP duration (MAPD90), ventricular late repolarization duration (VLRD) and intra- ventricular conduction time (IVCT) were prolonged by 26％ (P ＜ 0. 01), 43％ (P ＜ 0. 01), 318％ (P ＜ 0. 05), and 19％ (P ＜ 0. 01), respectively in CHF group. (2)The ratio of VERP to MAPD90(VERP/MAPD90) was decreased by 13％ (P＜0.05) in CHF group. (3) The dispersion of ventricular recovery time (VRT - D) was increased by 185％ (P ＜0. 01) in CHF group. (4) The ventricular fibrillation threshold (VFT) was decreased by 48％ (P ＜ 0. 01) in CHF group. CONCLUSION:The abnormal electrophysiological changes in the CHF condition may be contributing factors of lethal ventricular arrhythmias and sudden cardiac deaths in CHF.

BACKGROUND:The hyperpoladzation-activated cyclic nucleotide-gated cation channel (HCN) gene is not increase the risk of induced cardiac arrhythmia,and can accept the modulation function of autonomic nervous system.Therefore,it is the first candidate gene for biological pacemakers.OBJECTIVE:To construct recombinant adenovirus vector containing human HCN4 gene and evaluate its transfection efficency in rat bone marrow mesenchymal stem cells (MSCs).DESIGN,TIME AND SETTING:The in vitro cytology-gene experiment was performed at the Lin Bai-xin Experimental Center,Second Hospital of Sun Yat-sen University from February to September 2008.MATERIALS:Ten SD rats were supplied by the experimental animal center of Sun Yat-sen University.Rlasmid pcDNA3.1-HCN4 containing target gene human HCN4,human embryo kidney 293 cells and Escherichia coll DH5α were preserved by our experimental center;Shuttle plasmid pShuttle-CMV containing green fluorescent protein gene and adenoviral backbone plasmid pAdxsi were bought from SinoGenoMax Co.,Ltd.METHODS:HCN4 cDNA segment was liberated from the cloning vector of pcDNA3.1-HCN4 via Hind Ⅲ+Xba Ⅰ,and subcloned into pShuttle-CMV,which was digested by I-Ceu Ⅰ +I-Sce Ⅰ double enzyme and subcloned into adenoviral plasmid to form recombinant adenovirus plasmid.Recombinant adenovirus plasmid was transfected into 293 cell lines by liposome,and the recombinant adenovirus AdHCN4 was packaged and transfected into rat MSCs.MAIN OUTCOME MEASURES:The identification of recombinant adenovirus plasmid vector,identification of recombinant adenovirus and its titration test;the transfection efficiency of recombinant adenovirus.RESULTS:Cloned sequence about 3.6kbp was obtained by Hind Ⅲ+Xho Ⅰ digestion after HCN4 cDNA segment was cloned into pShuttle-CMV.DNA sequencing results indicated that the clone location was correct.Recombinant adenovirus plasmid was cut into seven fragments while empty vector gained only six fragments after digested by Xho Ⅰ.The recombinant adenovirus was pathogenic to 293 cells after recombinant adenovirus plasmid was packaged in it.HCN4 cDNA (657bp) was amplified by PCR with virus titer of 2.5×10~(11) PFU/mL after transfected 293 cells with supematant.The efficiency of recombinant adenovirus infecting rat MSCs was about 90% when multiplicity of infection was 800.Rat MSCs expressed green fluorescence after transfection.CONCLUSION:The adenovirus vector with human HCN4 cDNA was established successfully,which can effectively transfect rat MSCs.

The iodine concentration, MIT/DIT and T3/T4 ratios and peroxidase activity in thyroid of the rats fed on endemic goiter region diet of county Chefong in the Inner Mongolia Autonomous Region and control rats injected with 30% iodine oil 0.2 ml besides the same diet were investigated.The result demonstrated that the iodine concentration of thyroid was decreased with increasing of thyroid weight, and the MIT/DIT ,T3/T4 ratios in thyroid and thyroid peroxidase activity of the rats fed on endemic region diet were higher than that of control.These metabolic patterns showed that the rats fed on endemic region diet were in iodine deficient state and consistent with those rats fed on artificial low iodine diet.

AIM:To study the effects of FK506 binding protein 12.6(FKBP12.6) overexpression on the performance of sarcoplasmic reticulum(SR) in ventricular myocytes of rats with heart failure.METHODS:The adenovirus(Ad)-mediated gene transfer was used to overexpress FKBP12.6 in ventricular myocytes of rats with heart failure.Western blotting and reverse transcriptase-polymerase chain reaction(RT-PCR) analysis were used to reveale specific overexpression of FKBP12.6.X-rhod-1-AM was used as the Ca2+ indicator,and cells were viewed under a confocal microscope.RESULTS:Adenovirus mediated overexpression of FKBP12.6 resulted in a 5-fold increase in relative FKBP12.6 mRNA levels and a 4-fold increase in relative FKBP12.6 protein levels at 48 h after transfection compared with control.The amplitude of the fluorescence [Ca2+]i transient was significantly increased in Ad-FKBP12.6-GFP cardiomyocytes compared with Ad-GFP myocytes(peak F/F0,3.16?0.42 vs 1.43?0.38,P

Objective To explore the protective effect of folic acid supplementation in different ways on lipopolysaccharide-induced external malformations and skeletal malformations in mice.Methods The pregnant mice were divided into six groups randomly,including saline control group,FA control group,LPS group,LPS+FAig group,LPS+FAip group,and LPS+FAdw group.All dams were medicated on gestational day 8～12,and then sacrificed on gestational day 18,for each litter,the number of live fetuses,dead fetuses,resorption fetuses and external malformations of live fetuses were all counted.All fetuses were subsequently evaluated the skeletal malformations.Results A low dose of LPS injection during the second trimester resulted in external and skeletal malformations.Exencephaly and encephalomeningocele were two of the most common external malformations,skeletal malformations consisted mainly of the incompletion of supraoccipital ossification,sternal malformation and rib malformation.FA supplementation by three ways all attenuated the external and skeletal malformations.And the best protective effect was by oral administration.The incidence of external,sternal and rib malformation was all decreased.Conclusions Three ways can prevent the mice during the second trimester from the external and skeletal malformations caused by LPS injection,and the best protective effect was by intragastric administration.

Objective Implantable cardioverter defibrillator(ICD)can effectively treat lifethreatening ventricular arrhythmias.The most common side effect is inappropriate discharge.This study analyzes the incidence and causes of inappropriate discharges of ICD in our hospital.Methods Forty.threepatients implanted with ICD in our hospital from November 2001 to October 2007 were involved in our study.Patients were followed-up regularly.All episodes recorded and stored in the ICD were analyzed.Results Seven of the 43 patients underwent ninety-six inappropriate discharges.Inappropriate discharges in six patients were caused by supraventricular tachyarrhythmias(SVT).In one patient the discharge was caused by noise.Most inappropriate discharges occurred in the first year after implantation.The history of atrial fibrillation before implantation is an independent predictor of inappropriate discharges.Conclusions The incidence of inappropriate discharge is 16.3%in our study and the most common cause is SVT.Most inappropriate discharges occur in the first year after implantation.Patients with atrial fibrillation history have a higher risk of inappropriate discharges.

Objective To identify the imbalance of T cell-specific transcription factors T-bet/GATA-3,and to explore the modulation with dexamethasone and imiquimod in CD4+T cells from ovalbumin (OVA)sensitized mice.Methods CD4+T cells were obtained fromsingled-cell suspension of spleen(after lysis of RBC).ELISA assay was used to detect the concentrations of IL-4,IL-5 and IFN-γin superna tants and cell pellets,and the expression of T-bet and GATA-3 was detected by Western blot.Resuits In the control group,tIle low levels of IFN-γ were detected in the supernatants during 24 h.In OVA treatment group,the concentrations of IL-4,IL-5 were increased significantly,and the concentrations of IFN-γ were always low in the supernatants.In the dexamethasone treatment group,the concentrations of IFN-γ,IL-4 and IL-5 were all low in the supernatants during 24 h.In the imiquimod treatment group,the concentrations of IFN-γ were increased significantly,and the concentrations of IL-4 and IL-5 were decreased in the super natants.It worked at 6 h,and achieved the peak at 12 h,lasted over 24 h.In the control group,the expres sions of T-bet and GATA-3 were detected in CD4+T cells during 24 h.In OVA treatment group,the expressions of T-bet were decreased,and that of GATA-3 were increased rapidly in CD4+T cells.In dexam ethasone treatment group,the expressions of T-bet were always low in CD4+T cells,and that ofGATA-3 were no change during 24 h.In imiquimod treatment group,the expressions of T-bet were increased,andthat of GATA-3 were decreased in CD4+T cells.The protein expressions worked at 6 h.and achieved the peak at 12 h,lasted over 24 h.Conclusion The imbalance T cell-specific transcription factors T-bet/GA-TA-3 contributes to both high expression of GATA-3 and low expression of T-bet in CD4+T cells from OVA sensitized mice.Dexamethasone treatment inhibits the expression of T-bet in CD4+T cells and has no func tion in GATA-3.Imiquimod treatment modulates key master switches GATA-3 and T-bet that results in com mitting T helper cell to a TH 1 phenotype and imiquimod may play a key role in the regulation of TH2 cytokine responses in asthma.