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*

C57BL/6 mice were inoculated intranasally (50 microl) with serial 10-fold dilution of HAB/01 H5N1 virus. Three and five days later, three mice of each group were euthanized. Lung injury was assessed by observation of lung histopathology, virus titers and MCD50 were also measured. Our data showed that H5N1 viral infection in mice resulted in mainly epithelial injury and interstitial pneumonia, featuring significant weight loss, dramatically increased lung wet weight:body weight ratio, inflammatory cellular infiltration, alveolar and interstitial edema, hemorrhage in lungs with high virus titers, and MCD50 was 10(-6.5)/ 0.05 mL. These results suggested that a mouse model of H5N1 viral infection was successfully established which may benefit study of H5N1 avian influenza virus and pathogenic mechanism of host.
Animals ; Brain ; pathology ; virology ; Disease Models, Animal ; Female ; Humans ; Influenza A Virus, H5N1 Subtype ; pathogenicity ; Influenza, Human ; pathology ; virology ; Liver ; pathology ; virology ; Lung ; pathology ; virology ; Mice ; Mice, Inbred C57BL ; Random Allocation ; Spleen ; pathology ; virology

The aim of this study is to investigate the effect of hyperin on the cccDNA of duck hepatitis B virus and its immunological regulation. Duck hepatitis B virus (DHBV) infection model and normal mouse spleen lymphocyte were used to evaluate the anti-HBV and immunoregulation effects. The DHBV-DNA of serum was detected at different time points by using serum DOT-BLOT hybridization. Polymerase chain reaction (PCR) was used for the determination of nuclear covalent closed circular DNA (cccDNA). Cytokine secretion was determined by ELISA method. DHBV-DNA were inhibited by hyperin (25 or 50 mg x kg(-1)), while cccDNA of liver could be eliminated efficiently by hyperin (25 or 50 mg x kg(-1), P < 0.05, P < 0.01). The T helper 1 effector cytokine was markedly enhanced by hyperin (25 or 50 microg x mL(-1), P < 0.01). In conclusion, hyperin has anti-HBV activity via multiple targets and pathways, and cccDNA may be one of the important targets.
Animals ; Antiviral Agents ; pharmacology ; DNA, Circular ; metabolism ; DNA, Viral ; metabolism ; Hepadnaviridae Infections ; virology ; Hepatitis B Virus, Duck ; genetics ; Hepatitis, Viral, Animal ; virology ; Interferon-gamma ; secretion ; Interleukin-12 ; secretion ; Liver ; virology ; Lymphocytes ; secretion ; Mice ; Quercetin ; analogs & derivatives ; pharmacology ; Spleen ; pathology ; virology

Two strains of Avian leukosis virus subgroup B (ALV-B) were isolated for the first time in China Hy-line White on the cultured DF-1 cells which were inoculated tissue samples from by an ELISA assay, a histopathology examination and a PCR-based diagnosis. The results from the ELISA assay indicated that the positive rate of serum antibodies to ALV-B and ALV-J virus were 16.3% (15/92) and 13% (12/92), respectively. The histopathological examination indicated that two types of tumor cells existed at same focus in liver and spleen, which mainly were myelocytoma cells and lymphosarcoma cells. The PCR-based diagnosis were performed as follows: the cellular DNA was extracted from the inoculated DF-1 cells; the specific fragments of 1100 bp and 924 bp were obtained by a PCR system with the diagnostic primers of ALV-B and ALV-J; and the PCR results for ALV-A, MDV and REV were all negative. Then, the amplified fragments of the two ALV-B stains were partially sequenced and shown an identity of 92.8%,94.7% with the prototype strain of ALV-B (RSV Schmidt-ruppin B). The identities of two ALV-J strains with the prototype strain HPRS-103 at 96.9%, 91.5%; The identities of two ALV-J strains with the American prototype strain at 85.9%, 81.5%. Our study had shown that ALV-B was isolated for the first time from the ALV-J infected commercial layer flocks in China. It also indicated that the chance of genetic recombination among various subgroups of ALV was increased.
Animals ; Avian Leukosis ; pathology ; virology ; Avian Leukosis Virus ; classification ; genetics ; isolation & purification ; Cell Line ; Chickens ; China ; Liver ; pathology ; virology ; Molecular Sequence Data ; Phylogeny ; Poultry Diseases ; pathology ; virology ; Spleen ; pathology ; virology

Severe chronic active Epstein-Barr virus (EBV) infection is a rare and life-threatening illness. Although the criteria for diagnosis include chronic or recurrent infectious mononucleosis-like symptoms lasting more than 6 months and high titers of anti-EBV antibodies, clinical and laboratory findings may be heterogeneous and flexible application of those criteria is necessary in cases showing typical clinical and pathologic findings. We report a case of severe chronic active EBV infection in a 62-yr-old female patient who showed classical clinical findings with infiltration of EBV-infected T lymphocytes in the bone marrow, spleen, and lymph nodes, and died four months after presentation.
Antigens, CD3/biosynthesis ; Antigens, CD4/biosynthesis ; Antigens, CD8/biosynthesis ; Bone Marrow Cells/virology ; Epstein-Barr Virus Infections/*diagnosis/*mortality ; Female ; Herpesvirus 4, Human/genetics ; Human ; Immunohistochemistry ; Lymph Nodes/virology ; Lymphocytes/metabolism ; Middle Aged ; Organ Weight ; Spleen/pathology/virology ; Support, Non-U.S. Gov't ; T-Lymphocytes/virology

Studies were performed to determine the effects of Bcell suppression on the pathogenesis of Subgroup J avian leukosis virus (ALV-J) in broiler chickens. Neonatal chickens were treated with cyclophosphamide (CY) or PBS, and then infected with ALV-J (ADOL-7501) at 2 weeks of age. CY treatment induced B cell specific immunosuppression throughout the experiment confirmed by decreased bursal weight, intact lymphocyte mitogenetic activity stimulated by Con A and increased relative subpopulation of CD3-positive cells as measured by flow cytometry. Chickens in this experiment had Mareks disease virus exposure prior to three weeks of age as determined by the presence of lymphocytic infiltration and antibody. Virus neutralizing antibody against ALV-J was first observed at 6 weeks post-infection in some of the infected chickens in the PBS group. As expected, none of the chickens from the CY group and uninfected chickens developed virus-neutralizing antibody. The viremic status was measured by real time RT-PCR using SYBR green I dye. The percentage of viremic chickens was significantly higher, and more chickens had high titered viremia, in the CY treated group. No neoplastic foci consistent with ALVJ infection were observed in any of the experimental chickens. The frequency and intensity of viral antigen expression determined by immunohistochemistry was significantly higher in tissues from CY treated birds than those of PBS treated chickens at 3 weeks post-infection. This study showed that B cell specific immunosuppression with CY treatment in chickens resulted in increase in viremia and viral antigen load in tissues.
Animals ; Avian Leukosis/*immunology/virology ; Avian leukosis virus/genetics/*immunology ; Body Weight/physiology ; Bursa of Fabricius/immunology ; *Chickens ; Concanavalin A/immunology ; Cyclophosphamide/*pharmacology ; Flow Cytometry/veterinary ; Immunocompromised Host ; Immunohistochemistry/veterinary ; Immunophenotyping/veterinary ; Immunosuppressive Agents/*pharmacology ; Lymphocyte Activation/drug effects/immunology ; Organic Chemicals/chemistry ; Poultry Diseases/immunology/*virology ; RNA, Viral/chemistry/genetics ; Random Allocation ; Reverse Transcriptase Polymerase Chain Reaction/veterinary ; Spleen/immunology/virology ; Statistics, Nonparametric ; Viremia/veterinary

OBJECTIVETo explore the role of HIV-1 tat gene variations in AIDS dementia complex (ADC) pathogenesis.

METHODSHIV-1 tat genes derived from peripheral spleen and central basal ganglia of an AIDS patient with ADC and an AIDS patient without ADC were cloned for sequence analysis. HIV-1 tat gene sequence alignment was performed by using CLUSTAL W and the phylogentic analysis was conducted by using Neighbor-joining with MEGA4 software. All tat genes were used to construct recombinant retroviral expressing vector MSCV-IRES-GFP/tat. The MSCV-IRES-GFP/tat was cotransfected into 293T cells with pCMV-VSV-G and pUMVC vectors to assemble the recombinant retrovirus. After infection of gliomas U87 cells with equal amount of the recombinant retrovirus, TNF-α, and IL-1β concentrations in the supernatant of U87 cells were determined with ELISA.

RESULTSHIV-1 tat genes derived from peripheral spleen and central basal ganglia of the AIDS patient with ADC and the other one without ADC exhibited genetic variations. Tat variations and amino acid mutation sites existed mainly at Tat protein core functional area (38-47aa). All Tat proteins could induce U87 cells to produce TNF-α and IL-1β, but the level of IL-1β production was different among Tat proteins derived from the ADC patient's spleen, basal ganglia, and the non-ADC patient's spleen. The level of Tat proteins derived from the ADC patient's spleen, basal ganglia, and the non-ADC patient's spleen were obviously higher than that from the non-ADC patient's basal ganglia.

CONCLUSIONTat protein core functional area (38-47aa) may serve as the key area of enhancing the secretion of IL-1β. This may be related with the neurotoxicity of HIV-1 Tat.

AIDS Dementia Complex ; metabolism ; pathology ; virology ; Adult ; Amino Acid Sequence ; Basal Ganglia ; virology ; Cell Line, Tumor ; Gene Expression Regulation, Viral ; Genes, tat ; HIV-1 ; genetics ; pathogenicity ; Humans ; Interleukin-1beta ; biosynthesis ; genetics ; secretion ; Middle Aged ; Molecular Sequence Data ; Neuroglia ; pathology ; secretion ; Spleen ; virology ; Tumor Necrosis Factor-alpha ; biosynthesis ; genetics ; secretion ; tat Gene Products, Human Immunodeficiency Virus ; genetics ; physiology

OBJECTIVETo investigate the pathogenesis and immunogenicity of H9N2 influenza virus A/Guangzhou/333/99 (a reassortant of G1 and G9 viruses isolated from a female patient in 1999) in a mouse model of infection.

METHODSMice were infected with increasing virus titers. Viral load in the lungs and trachea was determined by EID50 assay. Pulmonary histopathology was assessed by hematoxylin-eosin staining. Anti-HI antibody titers and T-cell responses to viral HA were determined by ELISPOT and confirmed by flow cytometry.

RESULTSMice presented a mild syndrome after intranasal infection with A/Guangzhou/333/99 (H9N2) influenza virus. Virus was detected in the trachea and lungs of mice harvested on days 3, 6, and 9 post-infection. A T-cell response to viral HA was detected on day 6 and H9 HA-specific CD(4+) T-cells predominated. Seroconversion was detected after 14 days and antibody persisted for at least 28 weeks.

CONCLUSIONOur results suggest that H9N2 (A/Guangzhou/333/99) can replicate in the murine respiratory tract without prior adaptation, and both humoral and cell-mediated immunity play an important role in the immune response.

Animals ; CD4-Positive T-Lymphocytes ; immunology ; CD8-Positive T-Lymphocytes ; immunology ; Cell Line ; Dogs ; Enzyme-Linked Immunospot Assay ; Female ; Hemagglutination Inhibition Tests ; Hemagglutinins, Viral ; immunology ; Humans ; Infant ; Influenza A Virus, H9N2 Subtype ; immunology ; isolation & purification ; pathogenicity ; Interferon-gamma ; immunology ; Lung ; virology ; Lymphocytes ; immunology ; Mice ; Mice, Inbred BALB C ; Orthomyxoviridae Infections ; immunology ; virology ; Spleen ; immunology ; Trachea ; virology ; Viral Load ; Virulence

OBJECTIVETo investigate the correlation between the stage of hepatic fibrosis and ultrasonographic findings of the liver, spleen and gallbladder and establish a sensitive ultrasonographic semi-quantitative scoring system for screening compensated liver cirrhosis.

METHODSTotalling 248 patients with chronic hepatitis B and hepatitis C virus infection underwent liver biopsy and ultrasonic examination. The images of the liver surface, parenchymal echo, intrahepatic vessels, gallbladder, spleen and diameter of portal vein were analyzed.

RESULTSThe stages of hepatic fibrosis were not correlated to ultrasonographic findings of the liver surface or diameter of portal vein, but hepatic fibrosis of different stages showed significant differences in parenchymal echo, intrahepatic vessels, gallbladder and splenomegaly. In cases with normal liver parenchymal, intrahepatic vessels, gallbladder and spleen, the negative predictive value of the ultrasonographic semi-quantitative scoring system for diagnosing compensated liver cirrhosis amounted to 96.3%. The sensitivity of a score not lower than 5 was 90% for detecting compensated cirrhosis. With a score not lower than 7, the diagnostic accuracy and specificity was 85.9% and 95.2%, respectively, but the sensitivity was lowered to 37.5%.

CONCLUSIONThe ultrasonic images of the liver parenchyma, intrahepatic vessels, gallbladder and spleen in patients with compensated liver cirrhosis vary significantly in patients with hepatic fibrosis of different stages, and this ultrasonographic scoring system allows for a sensitive diagnosis of compensated cirrhosis.

Female ; Fibrosis ; Gallbladder ; diagnostic imaging ; Hepatitis B, Chronic ; complications ; Hepatitis C ; complications ; Humans ; Liver ; diagnostic imaging ; pathology ; virology ; Liver Cirrhosis ; complications ; diagnosis ; Male ; Reproducibility of Results ; Sensitivity and Specificity ; Spleen ; diagnostic imaging ; Splenomegaly ; diagnostic imaging ; Ultrasonography ; methods

OBJECTIVETo investigate the effect of Dureping Injection (DRP) on the T-cells function of mice and the function of T-cells in killing MF infected by influenza virus subtype A mice-lung adaptive strain FM1 in vitro.

METHODSNumber of splenic normal and FM1 infected T-cells in mice were measured by MTT and double-antibody sandwich ELISA, after being treated with DRP at different concentrations (2.1, 8.5 and 17.0 microg/mL), and the effect of DRP on interferon-gamma (IFN-gamma) and interleukin-10 (IL-10) production as well as on splenic T-cell killing FM1 infected Mphi/Ana-1 function were detected.

RESULTSDRP inhibited the multiplication of normal spleen T cells induced by concanavalin A in vitro, suppressed Th2 cell factor IL-10 production, and maintained Th1 cell factor IFN-y at a definite level, moreover, it directly enhanced the power of T-cells in killing FM1 infected Mphi (P < 0.05, P < 0.01).

CONCLUSIONDRP could act on mice T-cells to enhance the immune response for antiinfluenza viral FM1 in vitro.

Adjuvants, Immunologic ; pharmacology ; Animals ; Antiviral Agents ; pharmacology ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Influenza A virus ; drug effects ; Interferon-gamma ; immunology ; metabolism ; Interleukin-10 ; immunology ; metabolism ; Mice ; Mice, Inbred BALB C ; Orthomyxoviridae Infections ; immunology ; Spleen ; cytology ; T-Lymphocytes ; cytology ; immunology ; virology