Objective To detect antibodies against human papilloma virus-47 E2m345-362 peptide whichis homologous to profilaggri306324 peptide and anti-citrullinated human papilloma virus-47 E2345-362 peptide antibodies in rheumatoid arthritis(RA)and to investigate its role in the pathogenesis of RA.Methods Serum samples were obtained from 119 patients with RA, other rheumatic diseases and healthy individuals.We searched the homologus sequence of profilaggrin306-324peptide by using NCBI (the National Center for Bioteehnology Information)BLAST (basic local alignment search tool),and synthesized human papilloma virus-47 E2345-362 peptide which was highly homologous to profilaggrin306-324 peptide and the citrullinated Human papilloma virus-47 E2345-362 peptide.The presence of antibodies against E2345-362 peptide and citrullinated E2345-362 peptide was examined by enzyme-linked immunosorbent assay(ELISA).The associations between these antibodies and the clinical features of RA were evaluated.Results ①(41.2%)and titer (AU was 105.7)of anti citrullinated E2345-362 peptide antibodies in RA were significantly higher than those of patients with other rheumatic diseases and healthy individuals.However,the prevalence of anti-E2345-362 peptide antibodies in RA patients was similar to that of patients with other rheumatic diseases and healthy individuals(P>0.05).②The samples that were pre-incubated with cyclic citrullinated peptide (CCP) had lower titer of anti-citrulllinated E2345-362 peptide antibodies.③The titers of anti-CCP antibodies and anti-PAD14 antibodies in anti-citrullinated E2345-362 positive patients were higher than those of anti-citrullinated E2345-362 negtive patients.It showed significant correlations between anti-citrulllinated E2345-362 peptide antibodies and anti-PAD14 antibodies(r=0.485,P<0.01).④ DAS28 score,ESR,X-ray stages,AKA in anti-citrullinated E2345-362 positive patients were higher than those of anti-citrullinated E2345-362 negative patients.Conclusion The presence of anti-citrullinated E2345-362 peptide antibodies in RA indicates that HPV-47 E2 may act as an auto-antigen which may play an important role in the pathogenesis of RA.The increasing of PAD14 may make it easy for HPV-47 E2 to be citrullinated and may induce the subsequent auto-immune reactions.

Objective:To evaluate the effect of mucosal administration of altered collagen Ⅱ(CⅡ)263-272 peptide(267Q→A,270K→A and 271G→A) on collagen induced arthritis(CIA),and to explore the mechanism of the inhibitory effect of the altered CⅡ263-272 peptide on CIA.Methods:CIA was induced in Lewis rats by immunization with bovine CⅡ.Altered CⅡ263-272 peptide was given intranasally beginning from the onset of arthritis(100 ?g/dose,daily for 5 doses and continuing every other day for other 7 doses).Wild CⅡ263-272 peptide(100 ?g/dose) or PBS was administered as controls with the same procedure.Therapeutic effects were evaluated by arthritis scores,body weight change,and joint pathologic scores.The anti-CⅡ antibody and its subtypes were measured with ELISA.The cytokines of IFN-? and IL-10 were measured with ELISA.The induction of regulatory T cells was assessed by FACS analysis of percentage of peripheral CD4+CD25+ T cells,and by real-time PCR analysis of the expression of Foxp3 and TGF-? mRNA.Results:(1) Following treatment with the altered CⅡ263-272 peptide,arthiritis scores were reduced and body weight was increased.The mean arthritis scores of rats treated with altered peptide,wild peptide and PBS were 2.50?2.43,4.50?2.23 and 6.33?2.73,respectively.The altered peptide could retard the histologic lesion of the joints.(2) The titers of anti-CⅡ antibodies IgG and IgG1 in the three groups were similar,but the IgG2a in altered peptide-treated rats decreased markedly as compared with PBS-treated rats(0.56?0.19 vs 0.95?0.29,P

Tuberculosis is a chronic infectious diseases caused by Mycobacterium tuberculosis(MTB).As the drug-resistance characteristics are different in patients with various genotypes,thus,the gene polymorphism study have critical clinical significance.Among the all kinds of techniques,some have been used to analyze polymorphism for a long time and new development in that respect has also been made recently.On the other hand,some techniques are e-merging but demonstrate promising application prospects.This study summarizes the gene polymorphism study of MTB which have been used or are emerging in recent years,and points out a few shortcomings briefly.Our object is to make a contribution to theoretical basis and knowledge accumulation in the drug-resistance and epidemiological survey field.

Objective To investigate the clinical efficacy of white mustard seed moxibustion on four flower points [bilateral points Geshu(BL 17) and Danshu(BL 19)] in treating night sweat.Method One hundred and twenty patients with night sweat were randomly allocated to groups A, B and C, 40 cases each. Group A received white mustard seed moxibustion on four flower points; group B, oral administration of zuogui pills; group C, application of millet paste to four flower points. After 22 days of treatment, pre-treatment and post-treatment TCM symptoms and signs of hyperhidrosis scores were compared in the three groups and the clinical therapeutic effects were compared between the three groups.ResultThere was a statistically significant pre-/post-treatment difference in the TCM symptoms and signs of hyperhidrosis score in the three groups (P<0.05). The cure rate and the total efficacy rate were 50.0% and 92.5%, respectively, in group A; 27.5% and 75.0%, respectively, in group B; 12.5% and 50.0%, respectively, in group C. There were statistically significant differences in the cure rate and the total efficacy rate, and the TCM symptoms and signs of hyperhidrosis score between group A and group B or C (P<0.05) and between groups B and C (P<0.05).Conclusion White mustard seed moxibustion on four flower points is an effective way to treat night sweat and can markedly improve its symptoms in the patients.

Objective To revise the American College of Rheumatology classification criteria for rheumatoid arthritis(RA)with anti-cyclic citrullinated peptide(anti-CCP)antibodies and to evaluate its utility in the diagnosis of Chinese patients.Methods All patients from the Department of Rheumatology and Immunology of Peking University People's Hospital who had arthritis complaints in recent two years were enrolled.Patients were divided into RA group and non-RA group according to the clinical diagnosis by experienced rheumatologists.The diagnostic value of ACR criteria and the anti-CCP revised criteria(RA-6,RA-7 and RA-8)were evaluated by analyzing the clinical and laboratory parameters.Results A total of 604 patients were included in the study.312 patients were diagnosed as RA and 292 were diagnosed as other rheumatic diseases by rheumatologists.For those patients who had disease course for less than 2 years,the sensitivity of 1987 ACR criteria,RA-6,RA-7 and RA-8 criteria was 82.0%,91.0%.87.0%and 87.0%,respectively.The specificity of them was 95.6%,83.9%,95.6%and 95.6%.respectively.The sensitivity of 1987 ACR criteria,RA-6,RA-7 and RA-8 criteria for all the RA patients was 92.3%,96.8%.94.6%and 94.6%,respectively.The speciflcity of them was 92.8%,83.6%,92.8%and 92.8%.respectively.Conclusion The 1987 ACR criteria have high sensitivity and specificity in established RA.but its sensitivity in early RA is low.The RA-6criteria can improve the sensitivity dramatically but with reduced specificity.The RA-7 criteria can increase the sensitivity without sacrifice the specificity,especially in early RA patients.It may be used as a new set of classification criteria in clinical practice.

Objective To evaluate the prevalence and clinical significance of IgG,IgA and IgM isotypes of anti-peptidylarginine deiminase 4 (anti-PAD4) antibodies in early rheumatoid arthritis (RA).Methods IgG,IgA and IgM isotypes of anti-PAD4 antibodies were measured in the sera of 88 RA patients with disease duration less than 2 years,62 patients with other rheumatic diseases and 57 healthy subjects.The diagnostic performance of IgG,IgA and IgM isotypes of anti-PAD4 antibodies and their relationship with disease duration,DAS28,ESR,CRP,anti-CCP antibodies and rheumatoid factor (RF) were evaluated.Data analysis were performed using t test,U test and Spearman's association analysis.Results ① The sensitivities of IgG,IgA,and IgM isotypes for early RA were 28.41％,36.36％ and 9.09％ respectively.The specificities of IgG,IgA and IgM isotypes were 94.12％,93.28％ and 95.80％ respectively.② IgA isotype was positively correlated with age (r=0.234,P=0.028),DAS28 (r=0.309,P=0.007),ESR (r=0.382,P=0.000) and CRP (r=0.291,P=0.008),while negatively correlated with disease duration (r=-0.295,P=0.006).③ IgA isotype was positively correlated with IgG isotype (r=0.451,P＜0.01).In the IgG negative patients,the positivity of IgA isotype was 29％(18/63),which indicated that the IgA isotype might be helpful in diagnosing RA in IgG isotype negative patients.Conclusion Anti-PAD4 antibodies can be detected in early RA,primarily with IgA and IgG isotypes.IgA isotype has negative correlation with disease duration,indicating that IgA isotype of anti-PAD4 antibody may play a role in the very early stage RA.

Objective To detect influenza A virus by reverse transcription‐loop mediated isothermal amplification (RT‐LAMP) assay to established a rapid ,simple and visualization nucleic acid detection method .Methods The RT‐LAMP primers were designed in accordance with the hemagglutinin gene of influenza A virus .Then ,the specificity of the primers was evaluated by detection of different influenza viruses ,and the sensitivity was confirmed by testing multiple diluted RNA samples . Hydroxynaphthol blue (HNB) was used for visually evaluation and gel electrophoresis was used for validation .Clinical samples were detected by RT‐LAMP assay .Its consistency with fluorescent quantitative polymerase chain reaction (PCR) methods was compared .Results The primers of RT‐LAMP assay had high specificity . This technique could amplify influenza A virus accurately .The detection limit of RT‐LAMP assay was 2 .5 × 103 copies/mL by detection of multiple diluted RNA samples .In addition ,the results of RT‐LAM P assay could be visually inspected using HNB by color change ,and the results was in accordance with that of gel electrophoresis . RT‐LAMP assay was in consistence with fluorescent quantitative PCR when clinically applied .Conclusions RT‐LAMP assay is a rapid ,specific ,sensitive and simple method to detect influenza A virus .

Objective To introduce a kind of UPS for portable type-B ultrasonic in vehicular-charging at the situation of flowing in battle field. Methods Long time power supply for type-B ultrasonic and other related medical devices was realized through AC/DC alteration switch. Conclusion It functioned well in field test. Conclusion Being light and convenient with AC/DC, it is worth popularizing.

Objective To establish a novel method to detect autoantibodies against platelatespecific receptors by flow cytometric bead assay and study its clinical application. Methods The beads were coated with monoclonal antibodies SZ2, SZ22, SZ21 and 7E3 against platelet GP Ⅰ b, GP Ⅱ b, GP Ⅲa and GP Ⅱ b/Ⅲ a, respectively. Captured platelet glycoprotein and beads complex was detected by FITC labeled polyclonal goat antihuman immunoglobulin using flow cytometer. The platelet samples that reacted with antibodies (SZ2, SZ22, SZ21 and 7E3) negatively and positively were tested, respectively. Each sample was repeated 20 times to generate intra-day CV for the MFI and once a day for 8 days to generate inter-day CV values. The 85 ITP patients, 17 NITP patients and 50 controls from the First Affiliated Hospital of Soochow University during March 2006 to December 2008 were included in the studies. The sensitivity and specificity of these four platelet antibodies to diagnose ITP were analyzed using ROC curve. The results were compared with MAIPA. Results The CV of the intra-day-assay for samples negative to antibody SZ2, SZ22,SZ21 and 7E3 were 3.26%, 2. 86%, 1.65% and 4. 94%, respectively; While the CV of the intra-day-assay for samples positive to antibody SZ2, SZ22, SZ21 and 7E3 were 6. 16%, 4. 88%, 5.20% and 5. 85%,respectively. The CV of the inter-day-assay for samples negative to antibody SZ2, SZ22, SZ21 and 7E3 were 5. 86%, 4. 74%, 5.69% and 7.56%, respectively; While the CV of the inter-day-assay for samples positive to antibody SZ2, SZ22, SZ21 and 7E3 were 7.53%, 5.49%, 7.11% and 6.25%,respectively. The MFI for SZ2 in ITP group, NITP group and healthy control group were 1.49(0. 88-16. 24),1.12(1.00-1.33), 1.01 (0. 83-1.37), respectively, which showed significant differences (H = 36.89,P＜0.01). The MFI for SZ22 in the three groups were 1.55 (0.84-11.30), 1.13(1.03-1.29), 0.98(0. 85-1.24), respectively (H=28.41, P ＜0.01). The MFI of SZ21 were 1.50 (0.87-11.04), 1.13(0.97-1.32), 1.05 (0.85-1.48), respectively (H=54.42, P＜0. 01). The MFI for7E3 were 1.51(0. 84-9.81), 1.05(0.86-1.13), 1.03 (0.74-1.28), respectively (H =31.97, P ＜0.01). Based on ROC analysis, with cut-off values of 1.37, 1. 24, 1.48 and 1.28 for SZ2, SZ22, SZ21 and 7E3,respectively, the AUC were 0. 86, 0.90, 0. 87 and 0. 84, respectively. The sensitivities of the assays were 58. 82% (50/85), 52. 94% (45/85), 52.94% (45/85) and 51.76% (44/85), respectively. When all four antibodies were used, the sensitivity was increased to 74. 12% (63/85), which was higher than that of MAIPA [ 50. 59% (43/85) ,χ2 = 6. 78, P ＜ 0. 05) ]. Conclusion Flow cytometric bead assay can be used to detect four platelet-specific autoantibodies simultaneously, and may be a useful method to aid in the diagnosis of ITP.

Objective To investigate the effect of CXCL16 on the development of murine collageninduced arthritis (CIA). Methods CXCL16 mRNA of the involved synovium and serum CXCL16 protein were determined respectively by reverse transcription-polymerase chain reaction (RT-PCR) and enzyme linked immunosorbent assay (ELISA) in murine collagen-induced arthritis. The proliferation of lymphocytes from murine spleen and the level of RANKL mRNA, stimulated by CXCL16 at different concentrations (0,100, 200, 400, 800 ng/ml), was detected respectively by CCK-8 and RT-PCR, then the level of IL-2 and IFN-γ in culture supernatant was detected by ELISA. Comparisons between groups were tested by t test and one-way ANOVA analysis. Results The serum CXCL16 [(127± 10) vs (72±8) pg/ml, P＜0.05] and synovial CXCL16 mRNA (0.214±0.007 vs 0.375±0.009, P＜0.01) in CIA were all significantly higher than those in normal controls. The proliferation of CXCL16 (200, 400, 800 ng/ml) in CIA mouse lymphocytes, was significantly higher than that of CXCL16 (0 ng/ml) (0.51±0.06, 0.56±0.05, 0.55±0.04, (0.41±0.04, P＜0.05). And CXCL16 on the CIA stimulated lymphocyte proliferation was significantly higher than controls on normal lymphocytes (P＜0.05). Compared with blank control group, the expression of IL-2, IFN-γ and RANKL mRNA of CIA CXCL16 (400, 800 ng/ml) groups was higher significantly (P＜0.01). Conclusion CXCL16 plays an important role in the development of murine CIA by activating lymphocytes.