OBJECTIVE: To detect the Coxsackie virus B3(CVB3) gene in myocardium and spleen tissues in viral myocarditis(VMC) with sudden death and to explore the diagnostic method for VMC by means of seeking pathogene. METHODS: By in situ RT-PCR, the detection of CVB3 gene in myocardium and spleen sections were performed in sudden death group caused by VMC and non-cardiac death group. RESULTS: In VMC group, CVB3 gene-positive signals were seen in myocardium sections(3 out of total 8 cases, No. 1, 4, 7 cases) and spleen sections(4 out of total 8 cases, No. 2, 4, 6, 7 cases). In non-cardiac death group, no positive signals were detected in both myocardium and spleen tissues. CONCLUSION Positive detection of CVB3 gene in both myocardium and spleen maybe an important character of VMC and can improve the detecting pathogene in diagnosing VMC.
Death, Sudden ; Enterovirus B, Human/genetics* ; Heart/virology* ; Humans ; Myocarditis/virology* ; Reverse Transcriptase Polymerase Chain Reaction ; Spleen/virology*

Highly pathogenic avian influenza viruses (HPAIV) of the H5N1 subtype have spread since 2003 in poultry and wild birds in Asia, Europe and Africa. In Korea, the highly pathogenic H5N1 avian influenza outbreaks took place in 2003/2004, 2006/2007 and 2008. As the 2006/2007 isolates differ phylogenetically from the 2003/2004 isolates, we assessed the clinical responses of chickens, ducks and quails to intranasal inoculation of the 2006/2007 index case virus, A/chicken/Korea/IS/06. All the chickens and quails died on 3 days and 3-6 days post-inoculation (DPI), respectively, whilst the ducks only showed signs of mild depression. The uninoculated chickens and quails placed soon after with the inoculated flock died on 5.3 and 7.5 DPI, respectively. Both oropharyngeal and cloacal swabs were taken for all three species during various time intervals after inoculation. It was found that oropharyngeal swabs showed higher viral titers than in cloacal swabs applicable to all three avian species. The chickens and quails shed the virus until they died (up to 3 to 6 days after inoculation, respectively) whilst the ducks shed the virus on 2-4 DPI. The postmortem tissues collected from the chickens and quails on day 3 and days 4-5 and from clinically normal ducks that were euthanized on day 4 contained the virus. However, the ducks had significantly lower viral titers than the chickens or quails. Thus, the three avian species varied significantly in their clinical signs, mortality, tissue virus titers, and duration of virus shedding. Our observations suggest that duck and quail farms should be monitored particularly closely for the presence of HPAIV so that further virus transmission to other avian or mammalian hosts can be prevented.
Animals ; Antibodies, Viral/blood ; Brain/virology ; *Chickens ; *Coturnix ; *Ducks ; Heart/virology ; Influenza A Virus, H5N1 Subtype/*pathogenicity ; Influenza in Birds/epidemiology/transmission/*virology ; Kidney/virology ; Korea/epidemiology ; Lung/virology ; Virus Shedding

Animal model is very important for anti-EV71 (enterovirus 71) drug and vaccine development. 1-day-old suckling EV71 mouse model is the main in vivo model used in China. 1-day-old suckling EV71 mouse is too small to perform antiviral experiment. And the route of administration and dosage capacity are also restricted. A strong virulence EV71 virus strain was selected after screening from five EV71 strains with 1-day-old suckling mice. A mouse-adapted EV71 strain with increased virulence in 12-day-old suckling mice, EV71-M5, was generated after five serial passages of the parental EV71 strain in mice. Virus titers of EV71 infected mice heart, liver, spleen, lung, kidney, small intestine, brain and muscle tissue were determined by cytopathic effect (CPE) assay. The virus used in this model is the first isolated EV71 strain in China. And 2-week-old suckling mice were used in this model. This is a supplement for the EV71 animal model in China. Establishment of this EV71 model will provide an attractive platform for anti-EV71 vaccine and drug development.
Animals ; Disease Models, Animal ; Enterovirus A, Human ; isolation & purification ; physiology ; Enterovirus Infections ; Female ; Heart ; virology ; Intestines ; virology ; Mice ; Mice, Inbred BALB C ; Muscles ; virology ; Viral Load ; Virulence

BACKGROUNDRheumatic heart disease (RHD) is the most important sequela of rheumatic fever (RF): evidence that streptococcal infection is aetiological is prominent, but sometimes contradictory. Acute HSV-1 infection in mouse leads to carditis and valvulitis whereas recurrent infection results in inflammatory granulomatous lesions that resemble Aschoff bodies. Cells containing HSV-1 inclusions or virus infected giant cells appear similar to Anitschkow cells or Aschoff cells respectively. We hypothesized that HSV-1 infection also may be involved in RHD.

METHODSFormalin-fixed, paraffin-embedded valvular tissue samples from 32 patients with RHD were investigated for evidence of HSV-1 infection. HSV-1 antigen was detected by immunohistochemistry, using HSV-1-specific monoclonal and polyclonal antibodies. HSV-1 glycoprotein D gene sequences were amplified by nPCR, using beta-globin gene amplification in the same samples as internal control. Valvular tissue from 5 cases of sudden death and 3 cases died of neisseria meningitis without a history of valvular disease was used for comparison. HSV-1-infected lung tissue was used as positive control.

RESULTSHSV-1 antigens were detected in valvular tissues from 21 of 32 (65.6%) patients. Fifteen of these 21 (46.9% of cases), but no antigen-negative sample, were positive also for HSV DNA. Nucleotide sequence of PCR products was homologous to the targeted region of the HSV-1 glycoprotein D gene. HSV-1 antigen was present also in one case of sudden death but viral DNA was not found in any tissue sample from the comparison group. Results from reagent and positive controls were as anticipated.

CONCLUSIONSThis is the first study to show the presence of HSV-1 antigen and genomic DNA in valvular tissues from patients with RHD and provides evidence for an association of HSV-1 infection with some cases of rheumatic valvular disease.

Adolescent ; Adult ; Antigens, Viral ; isolation & purification ; DNA, Viral ; isolation & purification ; Female ; Heart Valve Diseases ; etiology ; virology ; Heart Valves ; pathology ; virology ; Herpes Simplex ; pathology ; virology ; Herpesvirus 1, Human ; immunology ; isolation & purification ; Humans ; Male ; Middle Aged ; Rheumatic Heart Disease ; pathology ; virology ; Viral Envelope Proteins ; genetics

A coxsackievirus B strain was successfully isolated by cells culture from cardiac muscle tissues of a dead Sichuan golden monkey with myocarditis from a zoo of Changchun in China. The isolate was consistent with CVB by morphology, physicochemistry test, animal regression test and RT-PCR. Analysis of VP1 partial gene sequence and detection of mice specific serum IgG showed that the strain isolated was a coxsackievirus B3. It was the first CVB case report in Sichuan golden monkey and the strain isolated was named CVB/SGM-05.
Animals ; Cercopithecus aethiops ; Enterovirus B, Human ; isolation & purification ; Haplorhini ; virology ; Heart ; virology ; Mice ; Reverse Transcriptase Polymerase Chain Reaction ; Vero Cells ; Viral Structural Proteins ; genetics

OBJECTIVETo explore the apoptotic pathway mediated by endoplasmic reticulum stress in the mouse myocardium with heart failure induced by acute viral myocarditis caused by B-3 Coxsackie virus.

METHODSForty BALB/c male mice were randomly divided into 2 groups (n = 20): the control group and the virus infection group. The BALB/c mouse myocarditis was induced by B-3 Coxsackie virus and the mouse behavior was observed conventionally. All the mice were sacrificed on day 7 and the changes of left ventricular pressure (LVP) and the rate of change of left ventricular pressure (LV dp/dt) were measured. The cardiomyocytic apoptosis was analyzed by TUNEL method and the mRNA expression level of endoplasmic reticulum haperones glucose-regulated protein (GRP)78 and GRP94 was detected by RT-PCR.

RESULTS(1) Compared with those of control group, the parameters of cardiac hemodynamics in the virus infection group were significantly decreased (P < 0.01); (2) Compared with that of control group, myocardial apoptosis was significantly increased in the myocardial cells from mice with heart failure induced by acute viral myocarditis (P < 0.01); (3) The mRNA expression level of GRP78 and GRP94 were increased significantly in the virus infection group compared with the control group.

CONCLUSIONThese findings suggest the endoplasmic reticulum stress may mediate the apoptosis of myocardial cells in the mice myocardium of heart failure induced by acute viral myocarditis caused by B-3 Coxsackie virus.

Animals ; Apoptosis ; Coxsackievirus Infections ; physiopathology ; Endoplasmic Reticulum Stress ; Heart ; physiopathology ; Heart Failure ; physiopathology ; virology ; Heat-Shock Proteins ; metabolism ; Male ; Membrane Glycoproteins ; metabolism ; Mice ; Mice, Inbred BALB C ; Myocarditis ; physiopathology ; virology ; Myocardium ; pathology ; Myocytes, Cardiac ; cytology

OBJECTIVETo explore the distribution of cytomegalovirus (CMV) in vascular tissues and the relationship between virus and atherosclerogenesis after CMV infecting mice.

METHODS(1) C57 BL/6J Murine model of CMV infection was established by intraperitoneal injection of CMV lethiferous amount. (2) After 12 weeks of CMV infection, the sera, carotids, aorta, hearts and postcaval veins from the mice were collected under euthanasia. The tissues would be used to DNA extraction, PCR and pathological examination. (3) Interleukin-6 (IL-6) and monocyte chemotactic protein-1 (MCP-1) in serum were measured with ILELISA.

RESULTS(1) The typical pathologic feature in 2 aorta samples of 6 mice infected by CMV was found and the mice uninfected by CMV did not show any pathologic change. (2) CMV DNA appeared in 6 aorta, 6 postcaval veins, 4 carotids and 4 heart tissues including endocardium, cardiac muscle and coronary artery from the CMV infected mice. CMV DNA was not found in the vascular and heart tissues from 6 mice uninfected by CMV. (3) The ELISA test showed the significant difference (Mann-Witney test of Nonparametric Test, P < 0.05) in serum IL-6 (Median among 25% and 75% percentile: 113.7 pg/ml vs. 49.77 pg/ml) and MCP-1 (Median among 25% and 75% percentile: 128.7 pg/ml vs. 45.36 pg/ml) between CMV infected mice and uninfected mice.

CONCLUSIONCardiovascular cells are CMV latent reservoir in host body and CMV infection and the cytokines induced by CMV infection probably relate to atherosclerogenesis.

Animals ; Atherosclerosis ; immunology ; pathology ; virology ; Blood Vessels ; immunology ; pathology ; virology ; Coronary Vessels ; immunology ; pathology ; virology ; Cytokines ; immunology ; Cytomegalovirus ; genetics ; isolation & purification ; Cytomegalovirus Infections ; immunology ; pathology ; virology ; DNA, Viral ; genetics ; Disease Models, Animal ; Female ; Heart ; virology ; Humans ; Mice ; Mice, Inbred C57BL ; Random Allocation

Group B coxsackieviruses (CVBs) are a group of common human pathogens producing various clinical symptoms. Although the virology of CVB is well known, there is limited information on viral pathogenesis and the relationship between clinical symptoms and viral phenotype, particularly for CVB type 2 (CVB2). In 2004 in Korea, two CVB2 strains were isolated: CB2/04/279 from stool of an acute myocarditis patient with heart failure and CB2/04/243 from an aseptic meningitis patient. In this study, a high degree of homology was observed between the CB2/04/279 and CB2/04/243 full genome sequences. The two Korean CVB2 isolates had 93.1% homology compared to 82.1%–82.5% nucleotide sequence identity with the cardiovirulence-associated reference CVB strain Ohio-1 (CVB/O). CVB2-induced pathogenesis was analyzed, focusing on virus-induced pathology of various tissues in 4-week-old BALB/c inbred male mice. Myocarditis developed and extensive pancreatic inflammation was observed in all mice infected with CB2/04/279 or CVB/O, but not in animals infected with CB2/04/243. This is the first report of the full-genomic sequence and pathogenesis of the CVB2 strain isolated from an acute myocarditis patient in Korea.
Animals ; Base Sequence ; Enterovirus ; Genome ; Heart Failure ; Humans ; Inflammation ; Korea* ; Male ; Meningitis, Aseptic* ; Mice* ; Myocarditis* ; Pathology ; Phenotype ; Virology

BACKGROUNDHepatitis B virus (HBV) replication has been reported to be involved in many extrahepatic viral disorders; however, the mechanism by which HBV is transinfected into extrahepatic tissues such as myocardium and causes HBV associated myocarditis remains largely unknown.

METHODSIn this study, endothelial progenitor cells (EPCs) were infected by HBV and then transfused into ischemic model of mice. HBV surface and core antigen as well as mutation of HBV particles were detected by immunohistochemistry, fluorescent activated cell sorter and transmission electron microscopy in vitro and in vivo.

RESULTSHuman cord blood EPCs, but not human umbilical vein endothelial cells (HUVECs) could be effectively infected by taking up HBV in vitro. HBV envelope surface and core antigen expressions were first detectable in EPCs at day 3 after virus challenge, sustained for up to 11 days, and decreased thereafter. Similarly, the virus particles were the most abundant in EPCs in the first week observed by a transmission electron microscope, and declined in 3 weeks after HBV infection. HBV DNA but not HBV cccDNA in EPCs were detectable even 3 weeks after virus challenge, as shown by PCR analysis. Furthermore, intravenous transplantation of HBV-treated EPCs into myocardial infarction Sprague & Dawley rats model resulted in incorporation of both EPCs and HBV into injured endothelial tissues of capillaries in the ischemic border zone.

CONCLUSIONSThese results strongly support that EPCs serve as virus carrier mediating HBV trans-infection into the injured myocardial tissues. The findings might suggest a novel mechanism for HBV-associated myocarditis.

Cell Movement ; Cells, Cultured ; Endothelial Cells ; cytology ; physiology ; Heart ; virology ; Hepatitis B virus ; physiology ; Humans ; Neovascularization, Physiologic ; Stem Cells ; physiology

OBJECTIVETo assess the diagnostic value of cardiac magnetic resonance (CMR) in patients with acute viral myocarditis.

METHODSThirty patients with suspected acute viral myocarditis admitted in first people's hospital of Shunde from June 2011 to June 2013 were included in this prospective study. The diagnostic sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and accuracy of acute viral myocarditis were evaluated by clinical diagnosis. Diagnostic value among different scan methods and Lake Louise criteria were compared.

RESULTSAcute viral myocarditis was diagnosed in 63.33% (19/30) patients.Values for sensitivity, specificity, PPV, NPV, and diagnostic accuracy within the overall cohort were 57.89%, 72.73%, 78.57%, 50.00%, 63.33%, respectively by edema imaging (ER).Values for sensitivity, specificity, PPV, NPV, and diagnostic accuracy within the overall cohort were 78.95%, 63.64%, 78.95%, 63.64%, 73.33%, respectively using global relative enhancement (gRE).Values for sensitivity, specificity, PPV, NPV, and diagnostic accuracy within the overall cohort were 78.95%, 54.55%, 75.00%, 60.00%, 70.00%, respectively using late gadolinium enhancement (LGE) criteria.Values for sensitivity, specificity, PPV, NPV, and diagnostic accuracy within the overall cohort were 84.21%, 81.82%, 88.89%, 75.00%, 83.33% using Lake Louise criteria. The sensitivity, specificity, PPV, NPV, and diagnostic accuracy using Lake Louise criteria were significantly higher than using ER, gRE, LGE alone(all P < 0.05).Specificity was higher using ER than using gRE and LGE (both P < 0.05). The sensitivity, NPV, and diagnostic accuracy were significantly higher using gRE than using ER (all P < 0.05) and was similar as using LGE (all P > 0.05).

CONCLUSIONCardiac magnetic resonance is an excellent imaging modality for the diagnosis of acute viral myocarditis.

Acute Disease ; Contrast Media ; Gadolinium ; Heart ; Humans ; Magnetic Resonance Spectroscopy ; Myocarditis ; diagnosis ; virology ; Predictive Value of Tests ; Prospective Studies ; Sensitivity and Specificity