Objective To elucidate the prevalence and clinical characteristics of human metapneumovirus(hMPV)in hospital- ized children with respiratory infection.Methods A total of 452 hospitalized children with lower respiratory tract infection were observed from Aug 2004 to Jan 2005.Respiratory tract aspirates were collected from all patients within 48 hours after admis sion.The specimens were routinely tested for respiratory syncytial virus,influenza virus A and B,parainfluenza virus 1 to 3 and adenovirus by direct fluorescent assay(DFA).The 245 specimens negative by DFA were tested for hMPV by RT-PCR. PCR products of hMPV M gene from some patients were randomly selected for sequencing analysis.Results hMPV was identi- fied in 59(24.1%)of the 245 specimens tested,hMPV infection alone accounted for 13.1% of the infections in the 452 chil- dren under study,The prevalence of hMPV was higher than other respiratory viruses in winter.The mean age of hMPV-infec- ted children(n=59)was 27.7 months.There was no significant difference between age groups in terms of the prevalence of hMPV(P＞0.05).There were no statistically significant difference in demographics and clinical symptoms between hMPV in- fection and other common respiratory virus infection.Genotyping for the hMPV M gene from 23 Shanghai patients showed two distinct hMPV genotypes.Sequence analysis of these hMPV M genes showed 82.8%-100% homology to the registered se- quence in GenBank.There was no significant difference in clinical characteristics between the 2 genotypes.Conclusions hMPV plays an important pathogenic role in lower respiratory tract infection of children,hMPV prevailed in the winter of 2004.Clini- cally,hMPV infection can not be discriminated from the infection of other respiratory viruses.Clinical manifestation is similar between the two hMPV genotypes.

OBJECTIVE: To study the forensic application of Goldeneye DNA ID 26Y Kit in the She nationality. METHODS: Through capillary electrophoresis, the genotype of 26 Y-STR loci were analyzed in 53 unrelated male individuals from Fujian She nationality. The population genetics parameters such as allele frequency and haplotype diversity were calculated. The comparisons among the She nationality and the other nationalities were analyzed. RESULTS: A total of 126 alleles were observed on the 26 Y-STR loci of 53 unrelated male individuals. The allele frequencies and GD value ranged from 0.010 1 to 0.886 8 and 0.211 2 to 0.846 2, respectively. The GD value was greater than 0.5 in the 19 loci. A total of 47 haplotypes were observed. Based on R(ST), multidimensional scaling plot indicated that the genetic relationship among Fujian She nationality and Minnan Han nationality was closest, followed by Southern China Han nationality and Northern China nationality. CONCLUSION Goldeneye™ DNA ID 26Y Kit including 26 Y-STR loci has good polymorphism in the She nationality. As an additional system, it has forensic application value in some special cases.
Asian People/genetics* ; China ; Chromosomes, Human, Y/genetics* ; Ethnicity/genetics* ; Forensic Genetics ; Genetic Markers ; Genetics, Population ; Humans ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Population Groups

Objective To find out the importance of human bocavirus (HBoV) as an infectious agent for population in Beijing, China. Seroprevalence study was conducted by using expressed recombinant major capsid VP2 protein as an antigen.Methods Serum specimens collected from infants and children who visited the Children's Hospital Affiliated to the Capital Institute of Pediatrics for health check-up and adults visiting the Xuanwu Hospital, Beijing for diseases other than respiratory infections from April 1996 to March 1997 were used for the investigation. The major capsid protein VP2 from HBoV was expressed in E. coli strain BL21 (DE3) with the transformed PET30b vector inserted with full-length VP2 gene of HBoV and the specific antigenicity of this expressed protein was validated by previous study. Western blotting was used to detect specific IgG antibody against HBoV in collected serum specimens diluted to 1:200. Mock expressed protein was E. coli cells strain BL21 (DE3) with the transformed PET30b vector without insert. Anti-His monoclonal antibody and rabbit anti-HBoV VP2 polypeptides hyper-immune serum were used as positive control for antibody detection.Results Out of 677 serum specimens tested, 400 (59.1% ) were positive for HBoV by Western blotting. About 45.3% (34/75) of the newborns under 1 month of age had anti-HBoV antibodies, and antibody positive rates were decreased in age groups of 1 and 2 months (41.4% and 31.3%, respectively) then increased in the following ages from 6 months to 7 years old ( from 45.6% to 69.7% ). The antibody positive rates were maintained at a relatively constant level ( about 70% ) in the age groups from 7 years to 40 years of age and became lower ( 61.8% - 62. 8% ) in those over 50 years.Conclusions The high seroprevalence of antibody against recombinant HBoV VP2 protein and early age antibody acquisition indicate that HBoV has been circulating in population of Beijing, China as early as in 1996 and most of children had been exposed to HBoV by the age of 7 years. Infants under the age of 6 months were susceptible to this virus.

OBJECTIVETo establish the triple-transgenic mouse model and study their biological characteristics by molecular biology, behavior and pathology.

METHODSHybrid the Tau and amyloid precursor protein (APP)/presenilins (PS1) transgenic mouse, the genotype of offspring mice were identified by PCR. Transcribed target genes were detected by RT-PCR. The protein expression of exogenous genes was detected by Western-blot. The pathological change of neurofibrillary tangles and senile plaque were observed by Bielschowsky silver staining and ABC immunohistochemical method. The changes time of learning and memory were observed by Morris water maze.

RESULTSAPP, PS1 and Tau genes were transcript in Tau/APP/PS1 mice. In 6 to 8 months old Tau/APP/PS1 mice, the neurofibrillary tangles and senile plaque could be found in cortex and hippocampus. In 6 months old Tau/APP/PS1 mice, the learning and memory abilities were worse.

CONCLUSIONWith the behavior change and pathological changes in Tau and beta-amyloid protein (AP), the Tau/APP/PS1 triple-transgenic mice can be used as a further study animal model of AD's pathogenesis and the target of drug treatment.

Alzheimer Disease ; pathology ; Amyloid beta-Protein Precursor ; genetics ; Animals ; Brain ; pathology ; Disease Models, Animal ; Learning ; Male ; Memory ; Mice ; Mice, Transgenic ; Neurofibrillary Tangles ; pathology ; Plaque, Amyloid ; pathology ; Presenilin-1 ; genetics ; tau Proteins ; genetics

OBJECTIVETo characterize the HA1 regions of hemagglutinin gene of influenza viruses (H3N2) isolated from children in Beijing from 1998 - 2004.

METHODSThe HA1 regions of hemagglutinin gene were amplified by RT-PCR from the viruses isolated and identified as A3 (H3N2) from clinical samples collected from infants and children during the peak seasons of influenza between 1998 and 2004. PCR products were sequenced or cloned into T-A vector and were analyzed after being sequenced.

RESULTSThe HA1 regions of hemagglutinin genes amplified from those isolates were 987 bp in length, encoding a protein of 329 amino acids in length. The identities of nucleotides and amino acids among these H3N2 isolates in Beijing and vaccines strains from 1998 - 2004 were 95.5% - 100.0% and 93.0% - 100.0%, respectively. The homology of the HA1 regions were related to the date of virus isolation, meaning the homology was higher among those strains isolated in nearer dates than others. Seven potential N-linked glycosylation sites in the HA1 regions located at amino acid positions 8, 22, 38, 63, 126, 165 and 285 were conserved in all the viruses analyzed. Two sites at 122 and 133 were inserted in those virus isolated after 1997, and another site at 144 appeared in those isolated after 1999. More amino acid substitutions located in the five putative antigenic sites or receptor binding sites were found more in the isolates than the isolates from previous year. Phylogenetic analysis showed new branches appeared continuously during 1998 - 2004. The strains isolated during winter in 2004 belonged to different branches, suggesting the appearance of new variants.

CONCLUSIONAmino acid substitutions continuously occurred in the HA1 regions of hemagglutinin genes in influenza virus (H3N2) isolated from children in Beijing from 1998 - 2004, which might have resulted in antigenic drift and led to the appearance of new variants.

Amino Acid Substitution ; China ; DNA, Viral ; analysis ; Gene Amplification ; Hemagglutinins ; genetics ; Humans ; Influenza A Virus, H3N2 Subtype ; genetics ; Influenza, Human ; virology ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA

BACKGROUNDHuman bocavirus (HBoV) is a newly identified human parvovirus that was originally detected in the respiratory secretions of children with respiratory infections. This study aimed to learn about the importance of HBoV infections by revealing the prevalence of serum antibodies against HBoV in Beijing population.

METHODSTwo batches of serum specimens collected in different periods were tested by Western blotting for specific IgG against HBoV using recombinant VP2 as antigen.

RESULTSOut of 677 serum specimens collected during April 1996 to March 1997, 400 (59.1%) were positive and antibody positive rate for another batch of 141 serum specimens collected in August, 2005 from adults aged from 20 years to over 60 years was 78.7% (111/141). Comparison of the sero-prevalence profiles for serum specimens collected during 1996 - 1997 to those collected in 2005 indicated that the antibody positive rate for specimens collected in 2005 was higher than that of the corresponding age groups collected during 1996 - 1997.

CONCLUSIONSThe data suggest that HBoV has been circulating in Beijing population for at least over 10 years, and most of children had been exposed to HBoV by age of 7 years. Higher HBoV antibody positive rate shown in the serum specimens collected in 2005 suggested that infections by HBoV have been increased in Beijing population in recent years.

Adult ; Antibodies, Viral ; blood ; Blotting, Western ; Bocavirus ; pathogenicity ; China ; epidemiology ; Humans ; Immunoglobulin G ; immunology ; Middle Aged ; Parvoviridae Infections ; blood ; epidemiology ; immunology ; Seroepidemiologic Studies ; Viral Proteins ; immunology ; Young Adult

OBJECTIVETo develop a rapid, sensitive and specific method in identifying and typing on adenovirus from clinical specimens.

METHODSPrimers were designed using hexon gene of adenovirus as target. One primer pair was designed as universal primers for amplifying a 1278 bp gene fragment located at the hexon gene of adenovirus from all types. Four primer pairs located within the region of this 1278 bp were specifically designed for amplifying types 3, 7, 11 and 21 of adenoviruses, which were used for multiplex nest-PCR in a single tube. The products from this multiplex nest-PCR were 502 bp (for type 3), 311 bp (for type 7), 880 bp (for type 11) and 237 bp (for type 21), respectively. Type of the adenovirus tested could then be determined after agarose electrophoresis analysis of the PCR products.

RESULTSPCR products with predicted sizes were visualized in the agarose gel for prototype strains of adenovirus types 3, 7, 11 and 21, but not for other respiratory viruses, indicating that the technique was specific without cross reaction with other viruses. Out of the 118 clinical specimens which had been proved to be adenovirus positive by tissue culture and/or immunofluerescence assay, 76 belonged to adenovirus type 3 (76/118, 64.4%), 37 to adenovirus type 7 (37/118, 31.4%), 3 to adenovirus type 11 (3/118, 2.5%) but no adenovirus type 21 was detected. Two of the 118 positive specimens which were positive by both tissue culture and immunofluerescence could not be identified, suggesting that these 2 strains (1.7%) were with the types other than types 3, 7, 11 and 21. Out of the 33 specimens which were negative by both tissue culture and immunofluerescence, 3 showed positive by this multiplex PCR (2 of type 3 and 1 of type 7), suggesting this method was more sensitive than tissue culture and immunofluerescence.

CONCLUSIONThis multiplex nest-PCR method had the benefit of rapid,sensitive and specific nature so could be used for identifying types of adenoviruses in the clinical specimens.

Adenoviridae ; classification ; isolation & purification ; Adenovirus Infections, Human ; virology ; DNA Primers ; DNA, Viral ; analysis ; Polymerase Chain Reaction ; methods ; Sensitivity and Specificity ; Sequence Analysis, DNA

OBJECTIVETo understand the relationship between human rhinovirus (HRV) and acute respiratory infections in infants and young children in Beijing.

METHODSThroat swab/nasopharyngeal aspirates were collected from 3292 infants and young children with acute respiratory tract infections in Beijing from November 2002 to November 2006. Primers derived from the highly conserved 5'-noncoding region of human rhinovirus were used to detect HRV from clinical specimens by nested RT-PCR for which the sensitivity and specificity had been determined previously.

RESULTSOut of these 3292 specimens, 507 were (15.4%, 507/3292) HRV positive with RT-PCR method. HRV were detected from 220 out of 1315 outpatients and 287 out of 1977 inpatients with positive rates as 16.7% and 14.5% respectively. HRV was detected from 50.0% (8/16) of the patients with pharyngitis. Among 280 specimens collected from patients with acute bronchitis, 43 (15.4%) were HRV positive, including 14 from 80 patients with wheezy bronchitis (17.5%). High positive rates were also found in specimens from patients with pneumonia (12.6%, 150/1189), bronchiolitis (16.0%, 42/262) and asthma (12.8%, 10/78). In 53 patients with initial diagnosis as hematic disease or other complicate respiratory infections, 14 were HRV (26.4%, 14/53) positive. As for the seasonal distribution, HRV were detected in most of the months during thie period of research. The highest positive rate of HRV in each year fell in September (32.6%), February (24.2%) of 2004, February of 2005 (35.3%) and March (31.3%) from 2003 to 2006, respectively. Among these HRV positive patients, 44.8% were under 1 year of age (227/507), 15.4% (78/507) were 1 to 2 years old and 12.4% (63/507) were 2 to 3 years old.

CONCLUSIONHRV was associated with acute upper respiratory infections and lower respiratory infections including bronchitis, pneumonia and bronchiolitis in pediatric patients. Patients with lower immunity such as those with hematic diseases, were more susceptible to be infected by HRV. HRV could be detected in all age groups in this study, but the positive rates were decreasing with the increase of patients' age. Infants under 1 year of age seemed to be more likely to get HRV infection.

Acute Disease ; Adolescent ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Picornaviridae Infections ; epidemiology ; virology ; Respiratory Tract Infections ; epidemiology ; virology ; Reverse Transcriptase Polymerase Chain Reaction ; Rhinovirus ; classification ; genetics ; pathogenicity ; Seasons

OBJECTIVETo develop a rapid, sensitive and specific method for detection human rhinovirus (HRV) from clinical specimens.

METHODSPrimers derived from the highly conserved 5'noncoding region of human rhinovirus were used to develop a nested RT-PCR for detecting HRV. The sensitivity and specificity of the RT-PCR were determined using various RNA while DNA viruses were used as control. Seven hundred and seventy-one specimens collected from children with symptoms of acute respiratory infections from Nov. 2002 to Oct. 2003 were analyzed for HRV by RT-PCR as well as for other respiratory viruses through isolation of virus and indirect immunofluorescent assay.

RESULTSOnly the cDNA from HRV was positive by RT-PCR, indicating the nested RT-PCR was specific. With RT-PCR, HRV were detected in 148 out of 771 specimens (19.2%). As for HRV positive rates, it was found 53.3% in pharyngitis patients; 43.8% in laryngitis patients and 28.7% in bronchitis patients. In Sep. 2002 and from Aug. 2003 to Oct. 2003, HRV positive rates were high (21.6% - 32.6%), with Sep. 2003 in particular--32.6%. From Mar. 2003 to Jul. 2003, HRV positive rates maintained from 16.0% to 19.1%.

CONCLUSIONHRV was one of the important agents for acute respiratory infections in infants and young children in Beijing.

5' Untranslated Regions ; Acute Disease ; Child, Preschool ; Conserved Sequence ; DNA, Viral ; analysis ; Humans ; Infant ; RNA, Viral ; analysis ; Respiratory Tract Infections ; virology ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Rhinovirus ; genetics ; isolation & purification ; Sensitivity and Specificity

OBJECTIVETo understand the relationship of parainfluenza virus (PIV) and acute respiratory infections in infants and young children in Beijing, occurred in recent years.

METHODS3141 throat swab/nasopharyngeal aspirate specimens were collected from infants and young children with acute respiratory tract infections in Beijing from Jan 2001 to Dec 2003. All of these 3141 specimens were inoculated into MDCK cells for influenza virus and PIV isolation, since PIV had been isolated in MDCK cells in this laboratory from preliminary studies. Out of 3141 specimens, 702 were inoculated into MDCK as well as Vero cells to compare the sensitivity on virus isolation of these cell lines by micro plate method. Growth of PIV in cell culture were identified by haemoagglutination test and indirect immunofluorescent assay.

RESULTSThe PIV positive cases in Vero cells were also positive in MDCK cells, indicating that the sensitivity for PIV isolation in MDCK was equal to Vero cells. Out of these 3141 specimens, 94 (3.0%) were PIV positive, including 35 (35/1191, 2.9%) of PIV1, 11 (11/1191, 0.9%) of PIV3 in upper respiratory tract infections; 15 (15/1634, 0.9%) of PIV1, 24 (24/1634, 1.5%) of PIV3 in lower respiratory tract infections; 3 (3/207, 1.4%) of PIV in asthma; 1 (1/38) of PIV in patients with fever; 5 (5/71) of PIV in others. Data indicated that among upper respiratory tract infections caused by PIV, PIV1 was more commonly seen than PIV3.

CONCLUSIONMDCK cells could be used for PIV isolation from clinical samples while PIV was one of the important pathogenic viruses causing acute respiratory tract infections in infants and young children in Beijing for the recente years.

Acute Disease ; Animals ; Cell Line ; Cercopithecus aethiops ; Child, Preschool ; China ; Humans ; Infant ; Parainfluenza Virus 1, Human ; isolation & purification ; pathogenicity ; Parainfluenza Virus 3, Human ; isolation & purification ; pathogenicity ; Paramyxoviridae Infections ; diagnosis ; Respiratory Tract Infections ; virology ; Sensitivity and Specificity ; Vero Cells ; Virus Cultivation ; methods