Objective To assess the diagnostic value of anti-mutated citrullinated vimentin antibodies (anti-MCV) for rheumatoid arthritis (RA), and compare it with anti-cyclic citrullinated peptide antibodies (anti-CCP), rheumatoid factors (RF). Methods Commercially available enzyme-linked immunosorbent assay (ELISA) kit was used to detect anti-MCV antibodies in a group of 177 RA patients, 46 patients with other rheumatic diseases, and 48 healthy blood donors. At the same time, anti-CCP, RF were detected. T test and χ2 test were selected. Results The average concentration of anti-MCV was (523±376) U/ml in RA, (96± 55) U/ml in patients with other rheumatic diseases, (34±18) U/ml in healthy controls. Different threshold levels (20, 40, 60, 80, 100, 120, 140 U/ml) for positive results were calculated bythe areas under the ROC curve (the areas were 0.521, 0.706, 0.769, 0.791, 0.816, 0.826, 0.822), then the best diagnosis efficacy for RA was determined as more than 120 U/ml. At this level, the sensitivity and the specificity for anti-MCV were 80.1% and 80.9% for RA diagnosis. The positive and negative predictive value were 92% and 67.8%. Comparing with anti-CCP, anti-MCV showed comparable specificity but higher sensitivity. And it's also better than RF apparently. If all 3 antibodies were detected at the same time, or anti-MCV combine with one of them, the sensitivity would increase to 95.7%. In addition, Anti-MCV showed positive in 32 of 67(55.2%) patients with RA whose anti-CCP was negative, meanwhile 31 of 59 (52.5%) patients with RA whose RF was negative. Conclusion RF and anti-CCP are complementary in diagnosing RA. The combination detection of RF and anti-CCP could significantly improve the specificity for the diagnosis of RA.

Objective To investigate the awareness rate of knowledge related with vascular cognitive impairment (VCI) in community residents in Shuangjing District,Beijing.Methods 5 communities were randomly selected from 12 communities in Shuangjing District.According to roster,512 residents were selected randomly.Among them,there were 197 males and 315 females with an average age of (64.71±8.22) years.87.9％ of subjects had junior high school culture and over.The awareness rate of knowledge related with VCI was investigated by questionnaires from January 2013 to March 2013.Results Although the awareness rate of knowledge related with vascular dementia was only 36.9％,the awareness rates of symptoms of impairment in memory,orientation,language,execution,calculation,visuospatial and judgement were 43.8％-89.3％.The awareness rates of four outpatient clinics among dementia patients were 21.5％-38.9％,however,the awareness rate of dementia which was not curable was 47.5％.The awareness rates of 5 risk factors for stroke were 54.5％-83.4％,but the awareness rate of cognition impairment caused by hypotension and hypoglycemia was 42.8％ and 43.2％,respectively.About VCI-related preventive knowledge,the awareness rates of VCI population with high risk factors,cognitive abilities screening,and primary prevention were 35.5％-95.5 ％.The main routes taking healthy knowledge were the television and newspapers with the awareness rate of 86.9％ and 60.2％,respectively.85.9％ of subjects agreed that healthy education was helpful to their life qualities.Multiple linear regression analysis showed a significant relationship of accumulated points of VCI-related knowledge with gender and education degree (P＜0.05).Conclusions The awareness rates of risk factors,diagnosis and therapy and primary prevention for VCI are lower in Shuangjing community,and the health education about VCI should be strengthened.

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.

ObjectiveThis work is aimed to investigate the possible association of dendritic cell immunoreceptor (DCIR) with rheumatoid arthritis (RA) susceptibility in Chinese Han population.Methods A total of 523 patients with RA and 510 healthy controls were genotyped for single-nucleotide polymorphism (SNP) rs2377422 and rs10840759.Association analyses were performed on the whole data set and on RA subsets based on the status of anti-cyclic citrullinated peptide antibody (CCP) in RA patients.Finally,we carried out the association analysis of rs2377422 with DCIR mRNA expression in RA patients.Statistical analysis used in this study included X2 test,Logistic regression,and Mann-Whitney U test.ResultsDCIR rs2377422 was found significantly associated with RA(allele analysis: OR 1.26; 95％CI 1.06～1.51,P=0.005; genotype analysis CC vs TT+TC: OR 1.34; 95％CI 1.18～2.06,P=0.004).Following stratification for anti-CCP antibody status,association of ra2377422 with anti-CCP-positive RA was observed(allele analysis: OR 1.22,95％CI 0.99～1.48,P=0.055).In contrast,the SNP rs2377422 was found specifically susceptible to anti-CCP-negative RA(allele analysis: OR 1.46; 95％CI 1.10～1.93,P=0.0091; genotype analysis CC vs TT+TC: OR 1.58;95％CI 1.01～2.47,P=0.043),despite loss of power in the analysis.DCIR gene transcription quantification analysis further proved the dominant effect of rs2480256 CC genotype on DCIR mRNA expression levels in RA patients (CC vs TT+TC: 0.429±0.069 vs 0.238±-0.023,U=1861,P=0.0015).ConclusionThe study provides evidence for the association between DCIR rs2377422 and RA,particularly with anti-CCP-negative RA in Chinese Han populations.

Objectives To evaluate the efficacy of rituximab combined with chemotherapy in the treatment of diffuse large B-cell lymphoma (DLBCL) and the relationship of clinical prognosis with the International Prognostic Index (IPI) by the using rituximab in autologous peripheral stem cell transplantation (APBSCT) for the patients of DLBCL. Methods 21 patients with DLBCL, 11 patients of them were at IPI low risk, and 3 patients were IPI at low intermediate risk, 3 patients were at IPI high intermediate risk, 4patients IPI high risk. Rituximab combined with CHOP regimen (cyclophosphamide, adriamycin, vincfistine and prednisone) was given for 4～8 courses. 5 patients received APBSCT. The mobilizing regimen was rituximab combined with cyclophosphamide(CTX) and etoposide(VP16). The conditioning regimen were CBV(CTX combined with VP16 and carmustine). Results In 21 patients, the complete response rate was 61.9 %,with overall response rate 90.5 %. 2-year progression free survival was (69.74±10.43)%. 2-year overall survival was (84.44:1:8.35) %. The complete response rate was 92.9 % and overall response rate was 100 % in the patients IPI≤2. The overall response rate was 71.4 % in the patients with IPI≥3. The complete response rate was higher in the patients with IPI≤ 2 (P＜0.01). The amount of mononuclear cells (M NC) in harvest were 7.34 (4.6～8.53)×108/kg. The CD+34 cells in harvest were 8.82 (2.1～10.34)×1O6/kg. The mean time of neutrephil recovering to 0.5×109/L after APBSCT was +9 day. The mean time of platelet recovering to 20×109/L after APBSCT was +12 day. The major adverse reaction were infusion related response (14.3 %) and hematological toxieities. Conclusion The efficacy of rituximab combined with chemotherapy in the treatment of DLBCL is effective, The complete response rate was higher in the patients with IPI≤2 than in the patients with IPI≥3.Using rituximab in mobilizing regimen, all patients had harvested enough CD+34 cells. Rituximab given at +1day did not affect the hematopoiesis reconstruction.

Objective To determine the expression pattern of macrophage-inducible c-type lectin (MINCLE)on Macrophage(Mφ),myeloid dendritic cell (mDC)and plasmacytoid DC(pDC)in peripheral blood (PB)and synovial fluid(SF)in patients with rheumatoid arthritis (RA).Methods For mRNA expression of MINCLE,253 RA patients and 71 healthy control subjects were enrolled.The mRNA level of MINCLE was determined by real-time PCR.For protein expression of MINCLE,18 patients with RA,5 patients with osteoarthritis(OA)and 12 healthy control subjects were enrolled.The expression of MINCLE on Mφ,mDC and pDC were detected by flow cytometry.The differences of MINCLE expressions in PB between RA patients,OA patients and healthy controls,or differences between PB and SF in RA patients were analyzed using Mann-Whitney U test or paired-samples t test.Results ①Compared to the healthy controls,RA patients showed elevated mRNA expression level of MINCLE in PBMCs[(1.65±0.36)vs (0.37±0.06),U=6057,P=2.75×10-5].②At protein level,MINCLE was hardly detected in Mφ,mDC and pDC in PB of OA patients and healthy controls.In SF,MINCLE was highiy expressed on mDC in RA patients,compared with that in OA patients[(34.8±4.4)%,U=0,P=2.6×10-3].In RA patients,the expression level of MINCLE was remarkably elevated in Mφ,mDC and pDC in SF compared with that in PB[Mφ(2.01±0.53)%vs(0.273±0.51)%,t=4.879,P=2.23×10-6;mDC(34.8±4.4)%vs(22.7±5.5)%t=2.535.P=0.017].Conclusion MINCLE is selectively expressed on Mφ.mDC and pDC in SF in RA patients.MINCLE may serve as a potential important marker,or even target,for RA and possibly even for inflammation in general.

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

OBJECTIVETo establish a rapid, sensitive and specific reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay for detecting human respiratory syncytial virus (RSV) in respiratory samples collected from children with acute respiratory infections.

METHODAccording to the conserved matrix gene sequences of respiratory syncytial virus subtypes A and B downloaded from GenBank, primers were designed and RT-LAMP assay was developed to detect RNA of RSV sensitivity of the RT-LAMP method was evaluated by using ten-fold serially diluted in vitro-transcribed matrix RNA fragments from RSV A and RSV B, respectively. Specificity of the RT-LAMP method was tested through cross-reaction with other RNA and DNA viruses. Then 5 RSV strains isolated from clinical specimens using tissue cultures were tested by RT-LAMP assay. A total of 101 nasopharyngeal aspirates from hospitalized patients with acute respiratory infections which had been tested by direct immunofluorescence assay (DFA), including 40 positive for RSV and 61 negative for RSV, were tested by RT-LAMP assay and by RT-nested PCR.

RESULTSensitivity analysis indicated that this RT-LAMP method was able to detect 1 copy/µl of RSV A and RSV B RNA, no amplification was shown in RT-LAMP with DNA or cDNA from other viruses in 60 min, revealed that the RT-LAMP assay is highly specific. Five RSV isolates confirmed as 4 RSV A and 1 RSV B previously were detected by RT-LAMP method as positive in 30 min. For those 101 specimens tested, 37 were RSV positive determined by RT-LAMP assay, as well as 35 RSV positive by RT-nested PCR. The total coincidence rate of RT-LAMP assay with DFA and RT-nested PCR in detecting RSV is 95.0%, 94.1% with Kappa value 0.895 and 0.871, respectively.

CONCLUSIONA new, sensitive, accurate and rapid method, RT-LAMP assay for detecting human respiratory syncytial viruses from nasopharyngeal aspirates was developed, which should be helpful in rapid detection of RSV from respiratory tract samples of children.

Acute Disease ; Child ; Child, Preschool ; DNA Primers ; Humans ; Infant ; Molecular Diagnostic Techniques ; Nasopharynx ; virology ; Nucleic Acid Amplification Techniques ; RNA, Viral ; isolation & purification ; Respiratory Syncytial Virus Infections ; diagnosis ; Respiratory Syncytial Virus, Human ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; Sensitivity and Specificity

OBJECTIVEThe present study was designed to explore the practical application of the rapid etiological diagnosis by detecting specific IgM antibody against common respiratory viruses in children with acute lower respiratory infections (ALRI).

METHODClinical specimens including nasopharyngeal aspirates and serum of acute phase from hospitalized children were collected from 207 infants and children with acute lower respiratory infections from March 2009 to September 2010. Seven common respiratory virus antigens were identified from the collected nasopharyngeal aspirates by direct immunofluorescence assay (DFA). ELISA was used to detect specific IgM antibody against RSV, ADV, IFVA, IFVB and PIV, while indirect immunofluorescence assay (IFA) was used to detect specific IgM antibody against RSV, ADV, IFVA, IFVB, PIV1, PIV2 and PIV3 in collected acute phase serum.

RESULTThe overall positive rates to detect viral antigen by using DFA, ELISA and IFA was 67.6%, 57.5% and 39.6%, respectively. The consistent rate of ELISA and IFA versus accepted DFA were 21.7% and 31.4%, respectively. The average days from onset of the symptoms to blood sample collection for those with the consistent results by ELISA and DFA were 12.0 d for ADV, 9.6 d for PIV2, 9.5 d for IFV, and 5.3 d for RSV, respectively, and by IFA and DFA were 15.0 d for PIV3, 9.2 d for ADV, and 7.4 d for RSV, respectively. Among all age groups, the consistent rate of serum viral IgM and antigen detections was highest in children younger than 3 years old.

CONCLUSIONAlthough there were differences between serum IgM antibody and viral antigen detections, specific IgM antibody detection was of value in early and rapid etiological diagnosis of pediatric ALRI, especially for young children. It could provide serologic evidence of respiratory virus infection. The diagnostic rate of pathogen could be improved if it was used in combination with viral antigen diagnostic methods.

Antibodies, Viral ; analysis ; blood ; Antibody Specificity ; Antigens, Viral ; analysis ; Child ; Child, Preschool ; Enzyme-Linked Immunosorbent Assay ; Female ; Fluorescent Antibody Technique ; Humans ; Immunoglobulin M ; analysis ; blood ; Infant ; Male ; Nasopharynx ; virology ; RNA Viruses ; genetics ; isolation & purification ; Respiratory Syncytial Virus Infections ; diagnosis ; virology ; Respiratory Syncytial Viruses ; genetics ; isolation & purification ; Respiratory Tract Infections ; diagnosis ; immunology ; virology ; Sensitivity and Specificity

Human metapneumovirus (hMPV) is associated with acute respiratory tract infections (ARTI) in all age groups. However, there is limited information of genetic analysis of hMPV circulating in Beijing. To learn the characteristics of structural protein genes of human metapneumovirus circulating in children in Beijing, sequence analysis of matrix (M), small hydrophobic (SH) and attachment (G) proteins of hMPV from 2006 to 2010 was performed. Phylogenetic analysis of nucleotide sequences of 42 full length M genes, 49 SH gene and 55 G gene revealed that the hMPVs from pediatric patients were divided into sub-genotypes A2, B1 and B. There were highly conserved identities among M gene, with 7 conserved mutations of amino acids between A and B genotypes which were fairly conserved in the same genotype A or B. The amino acid identities of SH were 60.7% to 64.4% between different genotypes, 93.3% - 100% among same sub-genotype and 84.7% - 88.7% between different sub-genotypes. Use of alternative transcription-termination codon, nucleotide deletion and insertion resulted in variable length of nucleotide and deduced amino acid of G protein. Amino acid identities within same genotype ranged from 81.5% - 100%, whereas sequence identities between two genotypes ranged from 34.0% - 38.6% at the amino acid level. A new cluster of G genes in sub-genotype B2 appeared due to the same mutations and insertion of two amino acids in G protein encoding genes amplified from specimens collected from 2008 to 2010. Prediction of antigen sites of SH and G protein indicated that the variation of antigen sites between different sub-genotypes existed.
Child ; China ; epidemiology ; Genetic Variation ; Genotype ; Humans ; Metapneumovirus ; genetics ; Paramyxoviridae Infections ; blood ; epidemiology ; virology ; Phylogeny ; Retroviridae Proteins, Oncogenic ; blood ; genetics ; Viral Envelope Proteins ; blood ; genetics ; Viral Matrix Proteins ; blood ; genetics