Fowl adenovirus serotype 4 (FAdV-4) strain SD1511 was isolated from chickens with severe inclusion body hepatitis and hydropericardium syndrome in Shandong Province, China. The isolate was cultured in primary chicken embryo kidney cells. A study of pathogenicity indicated that SD1511 readily infected 7-35-d-old chickens by intramuscular injection and intranasal and oral routes, causing 50%-100% mortality. The 35-d-old chickens suffered more severe infection than 7- and 21-d-old chickens with mortality highest in the intramuscular injection group. The serum from surviving chickens showed potent viral neutralizing capability. The complete genome of SD1511 was sequenced and analyzed. The strain was found to belong to the FAdV-4 cluster with more than 99% identity with the virulent FAdV-4 strains isolated in China in recent years except for some distinct variations, including deletions of open reading frame 27 (ORF27), ORF48, and part of ORF19. Our findings suggest that SD1511 might be used as a prototype strain for the study of pathogenesis and vaccine development.
Animals ; Antibodies, Neutralizing ; Aviadenovirus/pathogenicity* ; Cell Line ; Chick Embryo/virology* ; Chickens/virology* ; China ; Gene Deletion ; Genetic Variation ; Genome ; Genome, Viral ; Genomics ; Kidney/virology* ; Liver/virology* ; Open Reading Frames ; Poultry Diseases/virology* ; Serogroup ; Viral Load ; Virulence ; Virus Diseases/virology*

Objective: To understand the disease progression and human leukocyte antigen (HLA) gene polymorphism of HIV-infected persons without disease progress for long term, also known as long-term non-progressors (LTNPs), in Henan province. Methods: A retrospective study was conducted in 48 LTNPs with complete detection and follow-up information during 2011-2016 in Henan. Changes of CD(4)(+)T cells counts (CD(4)) and viral load (VL) during follow-up period were discussed. Polymerase chain reaction-sequence-specific oligonucleotide probe (PCR-SSOP) was used for the analyses of HLA-A, HLA-B and HLA-DRB1 alleles between LTNPs and healthy controls. Results: From 2011 to 2016, forty-eight LTNPs showed a decrease of the quartile (P(25)-P(75)) of CD(4) from 601.00 (488.50-708.72)/μl to 494.00 (367.00-672.00)/μl, and the difference was significant (P<0.05). The increase of the quartile (P(25)-P(75)) of log(10)VL from 3.40 (2.87-3.97) to 3.48 (2.60-4.37), but the difference was not significant (P>0.05). HLA polymorphism analysis revealed that HLA-B*13:02 and HLA-B*40:06 were more common in LTNPs (P<0.05), while HLA-B*46:01 and HLA-DRB1*09:01 were more common in healthy controls (P<0.05). Conclusions: The CD(4) of LTNPs in Henan showed a downward trend year by year. HLA-B*13:02 and B*40:06 might be associated with delayed disease progression for HIV infected persons in Henan.
Adult ; Alleles ; Asian People/genetics* ; China ; Disease Progression ; Female ; HIV ; HIV Infections/virology* ; HIV-1/immunology* ; HLA-B Antigens/genetics* ; Humans ; Middle Aged ; Polymorphism, Genetic ; Retrospective Studies ; Viral Load

OBJECTIVETo investigate the variations of hepatitis C virus (HCV) quasispecies and the changes in their composition in untreated patients with chronic hepatitis C.

METHODSEleven patients chronic hepatitis C without previous specific anti-HCV treatment were tracked for disease progression and blood samples were collected at multiple time points. The major clinical parameters of liver function and viral load were tested. A fragment of HCV hypervariable region 1 (HVR1) was amplified and cloned, and the positive clones were sequenced and subsequently analyzed to determine the composition variation of HCV quasispecies during disease progression in relation to the major clinical parameters.

RESULTSA total of 631 HVR1 sequences were acquired from the positive clones. The evolution of HCV HVR1 quasispecies in untreated chronic hepatitis C patients featured 3 patterns of variation in quasispecies composition, namely stable, fast and slow changes during the natural course of chronic hepatitis C. The genetic distance of the quasispecies was found to inversely correlated with ALT (R=-0.438, P=0.011) and AST level (R=-0.500, P=0.003), and sense mutation rate was also inversely correlated with ALT level (R=-0.387, P=0.026) and AST level (R=-0.410, P=0.018). No significant association was found between HCV load and any clinical or virological parameters.

CONCLUSIONDue to individual differences and immune pressure, HCV quasispecies can present with different patterns of evolution in the natural disease progression of chronic hepatitis C. HCV quasispecies evolution, due to its close correlation with the biochemical parameters, can be used to evaluate the severity and prognosis of chronic hepatitis C.

Base Sequence ; Disease Progression ; Genetic Variation ; Hepacivirus ; genetics ; Hepatitis C, Chronic ; virology ; Humans ; Viral Load

OBJECTIVETo study the relationship between metastasis or recurrence of hepatocellular carcinoma (HCC) and hepatitis B virus (HBV) DNA load or the presence of double mutation at 1762/1764 in the basic core promoter (BCP).

METHODSOne-hundred-and-fifty-seven patients with HCC were included in the study. Events of tumor metastasis or recurrence were recorded during 120 weeks of clinical follow-up after treatment by surgery or transarterial chemoembolization (TACE). The 1-year follow-up included monthly alpha fetoprotein (AFP) measurement and abdominal ultrasonography (US), as well as helical computed tomographic (CT) scan performed every 3 months. Follow-up beyond 1-year (surveillance) included AFP measurement and abdominal US every 2 months and helical CT scan every 6 months. Suspected metastasis or recurrence was investigated by hepatic angiography and confirmed according to the combined imaging findings. Serum HBV DNA level was measured by real-time PCR. HBV genotypes were determined by PCR-restriction fragment length polymorphism analysis.

RESULTSOf the 157 HCC cases 110 experienced tumor metastasis or recurrence; the cumulative probability of post-treatment HCC metastasis or recurrence was 4 (2.55%) at week 12, 14 (8.92%) at week 24, 28 (17.83%) at week 48, 64 (40.76%) at week 72, 92 (58.60%) at week 96, and 110 (70.06%) at week 120. Multivariate analysis indicated that both the BCP 1762/1764 double mutations and HBV DNA levels were risk factors for HCC recurrence or metastasis. In particular, the incidence of HCC recurrence or metastasis increased with baseline serum HBV DNA levels in a dose-response manner, ranging from 8/19 (42.1%) for less than 3 log10 copies/ml HBV DNA to 35/61 (57.3%) for 3-5 log10 copies/ml and 67/77 (87.0%) for more than 5 log10 copies/ml. After adjusting for potential confounders, serum HBV DNA level remained independently associated with HCC metastasis or recurrence. HCC recurrence or metastasis occurred in 22/43 (51.2%) of patients without BCP 1762/1764 mutations and 88/114 (77.2%) of patients with BCP 1762/1764 mutations. The adjusted odds ratio for patients infected with BCP 1762/1764 double mutation HBV, compared with those infected with non-BCP 1762/1764 mutation HBV, was 5.264 (95% CI: 1.436-12.574, P less than 0.05).

CONCLUSIONInfection with HBV carrying the BCP 1762/1764 double mutation and presence of high HBV DNA load are independent risk factors for developing HCC metastasis or recurrence after surgery or TACE.

Adult ; Aged ; Carcinoma, Hepatocellular ; pathology ; virology ; DNA, Viral ; blood ; Female ; Genotype ; Hepatitis B Core Antigens ; genetics ; Hepatitis B virus ; genetics ; Humans ; Liver Neoplasms ; pathology ; virology ; Male ; Middle Aged ; Mutation ; Neoplasm Metastasis ; Neoplasm Recurrence, Local ; Promoter Regions, Genetic ; Viral Load

OBJECTIVETo evaluate polymorphisms and forensic efficiency of 22 non-binary single nucleotide polymorphism (SNP) loci.

METHODSOne hundred ethnic Han Chinese individuals were recruited from Dongguan, Guangdong. The 22 loci were genotyped with matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS).

RESULTSNine loci were found with a single allele, 4 loci were found to be biallelic, whilst 9 loci were found to have 3 alleles. For 13 polymorphic loci, the combined discrimination power and power of exclusion were 0.999 98 and 0.9330, respectively. For the 9 non-biallelic loci, the combined discrimination power and power of exclusion were 0.9998 and 0.8956, respectively. For motherless cases, the combined power of exclusion was 0.6405 for 13 polymorphic SNPs and 0.6405 for 9 non-binary SNPs.

CONCLUSIONNon-binary loci have a greater discrimination power and exclusion power per SNP.

Alleles ; Asian Continental Ancestry Group ; genetics ; China ; Female ; Gene Frequency ; Genetic Load ; Genetics, Population ; Genotype ; Humans ; Male ; Polymorphism, Single Nucleotide

OBJECTIVETo investigate the genetic variation and typing of hepatitis B virus (HBV) in patients with chronic hepatitis B in relation to HBeAg status.

METHODSFluorescence quantitative polymerase chain reaction (PCR) was employed to detect serum HBV DNA in patients with chronic hepatitis B negative for HBeAg. Real-time fluorescent PCR and PCR-reverse dot blot hybridization were used to detect HBV genotypes and mutations, respectively.

RESULTSOf the 389 patients, 214 (55.01%) were positive and 175 (44.99%) were negative for HBV DNA; 102 (26.22%) had a HBV DNA copy number of 1×10(3), and 41 (10.54%) had a copy number of 1×10(4) (Χ(2)=226.6729, P<0.001). Of the 21 patients with a HBV DNA load of 1×10(5), 15 (71.43%) were found to have precore mutations, and 11 (52.38%) had basic core promoter (BCP) mutations; a higher HBV-DNA load was associated with an increased incidence of HBV mutations. In the 214 patients positive for HBV DNA, HBV genotypes A, B, C, D and the mixed type were found in 6 (2.80%), 84 (39.25%), 106 (49.53%), and 7 (3.27%), and 11 (5.14%) patients, who showed precore mutation rates of 16.67% (1 case), 36.90% (31 cases), 44.34% (47 cases), 0, and 0, and BCP mutation rates of 0, 19.05% ( 16 cases), 26.42% (28 cases), 0, and 0, respectively, demonstrating significant differences in HBV mutations between the genotype groups (P<0.001).

CONCLUSIONHBeAg-negative and HBV DNA-positive patients with chronic hepatitis B have a relatively low HBV replication level, and HBV DNA load is associated with HBV mutations. The B and C genotypes are more likely to have HBV mutations in HBeAg-negative patients.

DNA, Viral ; blood ; Female ; Genotype ; Hepatitis B e Antigens ; blood ; Hepatitis B virus ; classification ; genetics ; Hepatitis B, Chronic ; blood ; virology ; Humans ; Male ; Mutation ; Promoter Regions, Genetic ; Viral Core Proteins ; genetics ; Viral Load

A transient four-plasmid cotransfection system was used to construct avian influenza A (H5N1) pseudotyped viral particle (H5N1Pp) by incorporating hemagglutinin (HA) protein and neuraminidase (NA) protein from H5N1 avian influenza virus onto Murine leukemia virus pseudotyped viral particles, the transmission electron microscopy, infectivity titer assay, hemagglutination assay, neutralization assay of H5N1Pp were studied. We established a pseudotyped H5N1 viral particle at a high titer of 10(8) Pp/mL, the morphology,the hemagglutination activity and neutralization specificity of H5N1Pp is simililar to wild H5N1 virus. The research result sets a platform for studying this virus, including its receptors, the functional analysis of HA and NA, neutralizing antibodies and anti-H5N1 drug development.
Animals ; Birds ; Cricetinae ; Genetic Engineering ; HEK293 Cells ; Hemagglutination ; Hemagglutinin Glycoproteins, Influenza Virus ; genetics ; Humans ; Influenza A Virus, H5N1 Subtype ; genetics ; physiology ; Influenza in Birds ; virology ; Neutralization Tests ; Transfection ; Viral Load ; genetics ; Virion ; genetics

The purpose of this study is to elucidate the molecular mechanism of one-way serological reaction between XJ-160 virus and SINV by recombinant viruses which exchanged the glycoprotein genes individually or simultaneously. Three recombinant viruses were obtained based on the whole-length infectious cDNA clone of XJ-160 virus. The infectivity and pathogenesis to BHK-21 cells and animals were studied and the gene which controlled this one-way serological reaction phenomenon was searched by MCPENT. The results showed that the E2 glycoprotein was the main factor which influenced the growth rate, plaque morphology and pathogenicity of BHK-21 cells and suckling mice. The results of MCPENT showed that the E2 glycoprotein of SINV played a major role in this one-way serological reaction phenomenon. Our study identified the SINE2 gene was the determined gene for one way serological reaction between XJ-160 virus and SINV, and this research laid the foundation for further analysis of the genomic structure and function of SINV.
Alphavirus ; genetics ; immunology ; physiology ; Amino Acid Sequence ; Animals ; Cell Line ; DNA, Recombinant ; genetics ; Female ; Genetic Engineering ; Glycoproteins ; chemistry ; metabolism ; Mice ; Mice, Inbred BALB C ; Molecular Sequence Data ; Neutralization Tests ; Sindbis Virus ; immunology ; Viral Load ; Viral Proteins ; chemistry ; metabolism

OBJECTIVETo determine the complementary determining region 3 (CDR3) length diversity of T cell receptor Vbeta repertoires of CD8+ T lymphocytes and to explore its association with viral load in individuals with HIV-1 infection.

METHODSSeparation of CD8+ T cells from peripheral blood mononuclear cells (PBMCs) was carried out by using immunomagnetic beads coated with anti-CD8 antibody. Total RNAs from the purified CD8+ T lymphocytes were isolated and used to perform polymerase chain reaction (PCR) amplifications in CDR3 of 22 T cell receptor (TCR) gene families. CDR3 diversity and its association with viral load in individuals with HIV-1 infection were analyzed.

RESULTSAn average diversity for all CDR3 profiles in CD8+ T cells from 9 HIV-infected individuals was significantly different as compared to 7 age-matched healthy donors (P<0.05) with the HIV-infected individuals losing diversity in the CDR3 profiles. There was positive correlation between changes in TCR CDR3 diversity and viral load (r=0.771, P<0.05). The changes in CDR3 length diversity of Vbeta families in HIV-infected individuals, particular in Vbeta2, Vbeta4, Vbeta5, Vbeta17, Vbeta20, Vbeta21, Vbeta23, Vbeta24, were statistically different from the healthy controls.

CONCLUSIONHIV-1 infection might induce the loss of TCR Vbeta repertoire diversity and disrupt the CDR3 distributions within CD8+ T cells. There should be positive correlation between changes in TCR CDR3 diversity and the viral load in HIV-1 infected patients.

CD8-Positive T-Lymphocytes ; immunology ; HIV Infections ; genetics ; virology ; HIV-1 ; immunology ; Humans ; Polymorphism, Genetic ; Receptors, Antigen, T-Cell ; genetics ; Viral Load

OBJECTIVETo investigate the association of genetic polymorphism of dendritic cell-specific ICAM-grabbing nonintegrin (DC-SIGNR) and hepatitis C infection.

METHODSPatients with hepatitis C (n = 268) were genotyped and analysed for the repeat sequences polymorphism of DC-SIGNR using PCR and DNA sequencing. HCV virus load and HCV RNA genotypes were analyzed. Inter-group comparison was analyzed using LSD method.

RESULTSNo significant correlation was found between DC-SIGNR genotypes/ alleles and HCV RNA genotypes in patients. HCV-infected patients with 7-repeat (medium) alleles had lower HCV RNA levels compared to patients with 9-repeat (onger) alleles (P = 0.036). HCV-infected patients with 7/7 genotype had lower HCV RNA levels compared to patients with 9/7 genotype (P = 0.025). These findings suggest that optimal attachment of hepatitis C virions to DC-SIGNR may be associated with longer alleles.

CONCLUSIONThe fact that DC-SIGNR polymorphism might affect HCV loads supports the concept that DC-SIGNR contributes to HCV replication efficacy. There is no significant correlation between the genetic polymorphism of DC-SIGNR and HCV-RNA genotypes.

Adolescent ; Adult ; Aged ; Alleles ; Cell Adhesion Molecules ; genetics ; Child ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Hepacivirus ; genetics ; Hepatitis C, Chronic ; ethnology ; genetics ; virology ; Humans ; Lectins, C-Type ; genetics ; Male ; Middle Aged ; Polymerase Chain Reaction ; methods ; Polymorphism, Genetic ; RNA, Viral ; Receptors, Cell Surface ; genetics ; Viral Load ; Young Adult