OBJECTIVETo explore the effect of cytokine in the different prognosis of patients with severe influenza A (H1N1) infection.

METHODS28 cases with severe influenza A (H1N1) were enrolled in the study including 16 cured cases and 12 dead cases. The cytokine level in serum was detected by Luminex technology.

RESULTSThe levels of IL-2, IL-12 (P70) and IFN-gamma in dead group was lower than cured and normal control group and the difference were significant, P <0.05, respectively. IL-4 level in the dead group was significantly lower than cured group and normal control group, P value was 0.0310 and 0.0012, respectively.

CONCLUSIONSThe Thl cytokine level in the severe 2009 epidemic H1N1 influeaze cases shows decreased trend, and the trend is more obvious in dead cases. The decrease of Th1 cytokine may be one of reasons leading to severe clinical situation and related withthe bad prognosis.

Cytokines ; blood ; immunology ; Humans ; Influenza A Virus, H1N1 Subtype ; immunology ; isolation & purification ; Influenza, Human ; diagnosis ; immunology ; mortality ; virology ; Prognosis ; Th1 Cells ; immunology ; Th2 Cells ; immunology

OBJECTIVETo analyze the correlations between clinical features in paediatric patients with acute respiratory tract infection (ARTI) and viral load of human bocavirus.

METHODSA prospective study was conducted on 956 children < 5 years admitted with an acute respiratory tract infection from November 2009 to December 2010, and 251 healthy children conclused as control group in the corresponding period. Human bocavirus was investigated in nasopharyngeal aspirates (NPA) and throat swab by PCR, and viral load was detected by real-time polymerase chain reaction (RT-PCR) in HBoV positive sample. Clinical data were also prospectively recorded.

RESULTSA significant difference was found in HBoV positive rate between children with ARTI and control group at enrollment. There was a significant difference in HBoV viral load between children with upper respiratory tract infection and lower respiratory tract infection. HBoV viral load did not differ significantly between children with upper respiratory tract infection and control group. Among children with lower respiratory tract infection, no significant difference were detected between common and severe cases in HBoV viral load. HBoV viral load did not differ significantly whether the children were with or without co-infection.

CONCLUSIONSHBoV could be detected perennial and considered as a major pathogen associated with acute respiratory tract infection in children. However, HBoV may not be a independent factor in children with ARTI and the HBoV viral load was not associated with the severity of respiratory illness.

Case-Control Studies ; Child, Preschool ; Female ; Human bocavirus ; genetics ; isolation & purification ; physiology ; Humans ; Infant ; Male ; Parvoviridae Infections ; virology ; Respiratory Tract Infections ; virology ; Viral Load

OBJECTIVEIn an attempt to study the moleculr characterization and epidemiology of simian adenoviruses in nonhuman primate (NHP) populations.

METHODSWe examined a colony of captively bred rhesus macaques (Macaca mulatta) in China for the presence of adenoviral DNA in stool samples. This was done by using the PCR method that targeted the adenovirus polymerase gene, and the PCR positive fragments were cloned for sequencing and phylogenetic analyses.

RESULTSAmong the 57 animals analyzed, fecal samples from 12 animals were positive for the presence of adenoviral DNA. The results suggested that the viral DNA clones were primarily segregated into two large groups: SAdV-6 (2 non-redundant sequences) and SAdV-7 (9 non-redundant sequences). In addition, there were three clones with more similarity to SAdV-1, SAdV-3 and HAdV-52 respectively.

CONCLUSIONOur data confirmed the prevalence of adenoviral DNA in the feces of NHPs and revealed the adenoviruses in the gastrointestinal tract of the study animals. heterogeneity and phylogenetics of the adenoviruses in the gastrointestinal tract of the study animals.

Adenoviridae ; classification ; enzymology ; genetics ; isolation & purification ; Animals ; DNA-Directed DNA Polymerase ; genetics ; Feces ; virology ; Female ; Macaca mulatta ; virology ; Male ; Molecular Sequence Data ; Phylogeny ; Sequence Analysis, DNA ; Viral Proteins ; genetics

OBJECTIVETo investigate the correlation between the expression of tumor O (6)-methylquanine DNA methyl-tranferase(MGMT) and pathological grade,and the influence of racial factors on tumor MGMT expression levels for glioma patients.

METHODSCompare and analysis the correlation between the pathological grade and MGMT levels and the racial factors on MGMT expression levels by the immunohistochemical staining on the tumor specimens of 33 Uygur and 61 Han.

RESULTSThe positive rate of 61 Han gliomas pations with MGMT is 45.90% and 33 cases of the Uygur is 30.30% , there's no clear correlation between the racial factors and the tumor MGMT levels. (P >0.05). Comparative the 94 patients with pathological level and tumor MGMT level, there is no clear correlation between pathologic level and MGMT pression in tumor tissues (P >0.05).

CONCLUSIONThere's no clear correlation of tumor MGMT expression and pathological levels; and there's no significant effect between racial factors and expression of glioma MGMT.

Adolescent ; Adult ; Aged ; Child ; China ; ethnology ; DNA Modification Methylases ; genetics ; metabolism ; DNA Repair Enzymes ; genetics ; metabolism ; Female ; Gene Expression Regulation, Enzymologic ; Gene Expression Regulation, Neoplastic ; Glioma ; enzymology ; ethnology ; genetics ; pathology ; Humans ; Male ; Middle Aged ; Tumor Suppressor Proteins ; genetics ; metabolism ; Young Adult

OBJECTIVETo investigate human cytomegalovirus (HCMV) glycoprotein B (gB) genotypes and clinical features in neonates with congenital infections.

METHODSUrine samples were obtained from 67 neonates with HCMV infection confirmed by polymerase chain reaction (PCR). The gB gene fragment was amplified by nested PCR. HCMV gB genotyping was detected by restriction fragment length polymorphism.

RESULTSIn all these cases, the most prevalent genotype was gBl (50.7%), followed by gB3 (23.9%), gB2 (17.9%), and gBl/gB3 coinfection (7.5%); gB4 was not found. Moreover, gB1 was more prevalent in infants with liver damage (27/37, 73.0%) than in other symptomatic infants without liver damage (13/30, 43.3%; P < 0.05).

CONCLUSIONThe gBI genotype is the most prevalent in infants with congenital symptomatic HCMV disease, especially in those with liver damage, followed by genotypes gB3, gB2, and gB4.

Cytomegalovirus ; genetics ; isolation & purification ; Cytomegalovirus Infections ; congenital ; urine ; virology ; Female ; Genotype ; Humans ; Infant ; Infant, Newborn ; Infant, Newborn, Diseases ; virology ; Male ; Viral Envelope Proteins ; genetics ; urine

OBJECTIVETo analyze the molecular characteristics of the newly isolated two Japanese encephalitis virus strains (JEV) in Wuhan.

METHODSThe mosquitoes were collected in Wuhan from April to October in 2009. The envelope (E) protein gene of JEV was detected using RT-PCR and sequenced. Sequence comparisons and phylogenetic analysis were conducted using DNAstar and MegAlign.

RESULTSTwo Japanese encephalitis virus (JEV) strains (WHJX09-9, WHJX09-10) were isolated from Culex tritaeniorhynchus among 16 mosquito pools and identified as genotype I. The result showed that the homology of the two strains was 98. 9% in nucleotides and 100% in deduced amines. The comparison between the new genotype 1 JEV strains and live attenuated vaccine strain SA14-14-2 in E gene showed that the homology of nucleotide sequence was 87.4% and 87.9%, the homology of amino acid was 96.9% (total 15 amino acid were different) in E gene. The mutation sites of amino acid distributed among three different coding domain, but no antigen binding site and neurotoxin-involved site of amino acid were changed.

CONCLUSIONWuhan had appeared a new genotype of JEV which was different from the former strain isolated in Wuhan, the new JEV strains still had neurotoxicity but had high homology with the vaccine strains adopted in Wuhan. The vaccine could still be adopted to prevent Japanese encephalitis if steps were take to eradicate mosquitos at the same time. laboratory surveillance were also an important task to build an early-warning mechanism against JEV.

Amino Acid Sequence ; Animals ; Cell Line ; China ; Culicidae ; virology ; Encephalitis Virus, Japanese ; chemistry ; classification ; genetics ; isolation & purification ; Genotype ; Insect Vectors ; virology ; Molecular Sequence Data ; Phylogeny ; Sequence Alignment ; Viral Envelope Proteins ; chemistry ; genetics

OBJECTIVETo study the diversity of HIV-1 tat gene in CNS and peripheral tissue of a patient with ADC and a patient with non-ADC, so as to research HIV evolution, the mechanism of CNS invasion and the pathogenesis of ADC.

METHODSThe tat gene was amplified with nested PCR from genomic DNA which was extracted from spleen and basal ganglia of one non-ADC patient with a wide range of cerebral artery atherosclerosis and one ADC patient. PCR products were cloned into the PGEM-T vector, after transformation and selection by ampicillin and blue/white spotting. Five of positive clones were sequenced. HIV-1 tat sequences were processed with BioEdit and MEGA4. With the softwares, neighbor-joining tree, p-distances, values of ds/dn, and analysis of amino acid motifs were all done, so as to research the diversity of HIV-1 tat gene in CNS and peripheral tissue.

RESULTSGene mutation of HIV-1 tat exist in the two patients, the mutation process of tat isolated from ADC patient suffered more compartmentalization than tat isolated from non-ADC patient, the differences of tat genes between CNS and peripheral tissue in ADC patient were greater than the non-ADC patient. Ds/dn showed that the virus gene mutation played a major role, the body intend to remove harmful non-synonymous mutations.

CONCLUSIONSThe compartmentation of tat gene in CNS and peripheral tissue of the two patients was different, the reason may be related to the pathway of HIV into the CNS, the relationship between HIV gene mutation in CNS and ADC still need more investigation.

AIDS Dementia Complex ; virology ; Adult ; Amino Acid Sequence ; Central Nervous System ; virology ; Female ; Genetic Variation ; HIV-1 ; genetics ; isolation & purification ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Peripheral Nervous System ; virology ; tat Gene Products, Human Immunodeficiency Virus ; genetics

OBJECTIVETo compare the difference of HBV DNA levels and HBV genotypes between the patients with primary hepatocellular carcinoma (HCC) and liver cirrhosis who infected with hepatitis B virus.

METHODSTotal 430 patients with hepatitis B were enrolled and further divided into the HCC group (210 cases) and liver cirrhosis group (HBV LC, 220 cases). The levels of HBV DNA and HBV genotypes were detected in all of the serum samples from the two groups, and the differences in the genotypes and virological markers between HCC patients and HBV LC patients were further analyzed.

RESULTSThe positive rates of HBV DNA of HCC patients and HBV LC patients were 84.3% (177/210) and 94. 5% (208/220), respectively. The mean values of serum HBV DNA in HCC patients and HBV-LC patients were (5.06 +/- 1.01) log10 cps/ml and (5.36 +/- 1.13) log10 cps/ml, respectively. The positive rates of HBV DNA and the mean values of serum HBV DNA were higher in HBV-LC patients than those in HCC patients (P < 0.01). Furthermore, the main genotype was C in both groups and the distribution of genotype C and genotype B had no statistical difference.

CONCLUSIONSMainly presented as a C genotype in both groups, the total levels of serum HBV DNA in HCC patients were lower than those in HBV-LC patients.

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Hepatocellular ; blood ; virology ; DNA, Viral ; blood ; genetics ; Female ; Genotype ; Hepatitis B ; blood ; virology ; Hepatitis B virus ; genetics ; isolation & purification ; Humans ; Liver Cirrhosis ; virology ; Liver Neoplasms ; blood ; virology ; Male ; Middle Aged

OBJECTIVETo explore the variations of gene C in hepatitis B viruses between hepatitis B patients and healthy carriers, and provide experimental evidences for analysis of virus gene mutations acting on the virus material science and response of the body to the virus.

METHODSThe virus DNA load in hepatitis B patients and healthy blood donors was investigated by real-time polymerase chain reaction (PCR). Gene sequence analysis was taken to detect gene polymorphism, and all the success samples were compaired with standard strain by DNAstar.

RESULTS(1)G Compared with standard strain, C region in all samples had mutations, there were 31 mutations in at least 2 samples (3 mutations in gene PreC and 28 mutations in gene C), including 9 missense mutations, 1 chain termination mutation and 21 synonymous mutation. Mutations nt 1827 c-->a and nt 2221 c-->t existed in all the samples, and most samples had 6 synonymous mutations. Four hepatitis B patients had mutation nt1896 g-->a, and another 4 patients had 2 mutations, namely, S87G and I97F (or 197L) in HBcAg CTL recognition episome. (2) The success ratio of amplification and sequencing of HBV DNA was closely associated with its copy numbers. In the present study, copy numbers of HBV DNA which were successfully amplified and sequenced were almost more than 40 193/ml.

CONCLUSIONSHBV genome were easily affected by nucleotide mutations, 2 residues had mutations in gene of C region, which is firstly reported, suggesting these mutations may be geographical restricted. Mutations in gene of C region may either change the structure and function of HBeAg and HBcAg, which may further induce the escape of immune clearance for HBV or influence the detection of HBsAg or HBeAg, which may creat new problems for the prevention, diagnosis and treatment of hepatitis B.

Female ; Hepatitis B ; virology ; Hepatitis B Core Antigens ; genetics ; Hepatitis B virus ; genetics ; isolation & purification ; Humans ; Male ; Mutation ; Polymorphism, Genetic

OBJECTIVETo prepare streptavidin-tagged hepatitis C virus (HCV) fusion protein and explore its application for the detection of antibody against HCV infection.

METHODSA recombinant plasmid pET-11d-C44P-SA was constructed, which coding a novel HCV diagnostic antigens (C44P) and streptavidin (SA) fusion protein, and the fusion protein was generated with BL21 (DE3) E Coli and identified by Western Blot analysis. Then the fusion protein was purified through the Ni-NTA affinity chromatography and over 90% purity has been achieved. Anti-HCV ELISAs were developed when the fusion protein was used in the biotin-pre-coated microplate or ordinary microplate, and then the sensitivity and specificity of the ELISA were evaluated with confirmed human sera panels.

RESULTSThe fusion protein was expressed in high yields and purified successfully, the ELISA detection of anti-HCV with human sera panel indicated that its sensitivity and specificity is higher when SA-tagged HCV antigen (C44P-SA) coated in biotin-pre-coated microplate, compared to C44P or C44P-SA coated in ordinary microplate.

CONCLUSIONThe sensitivity and specificity of anti-HCV ELISA can be improved when a novel HCV diagnostic antigen fused to SA combined with the biotin- pre-coated microplate. This study laid a foundation for improving the performance of HCV diagnostics.

Antibodies, Viral ; immunology ; Enzyme-Linked Immunosorbent Assay ; Hepatitis C Antigens ; genetics ; immunology ; metabolism ; Recombinant Proteins ; genetics ; immunology ; metabolism ; Streptavidin ; genetics ; immunology ; metabolism