Development and clinical application of male human papillomavirus genotyping by membrane DNA chip / 中华男科学杂志

<p><b>OBJECTIVE</b>To develop a new method for the detection of male human papillomavirus (HPV) genotypes and to investigate its clinical application value.</p><p><b>METHODS</b>With computer assistance and based on the classical common primers MY09/11, modified PGMY09/11 with 23 HPV subtypes for PCR and Genbank data on HPV, we designed probes for the simultaneous detection of 18 high-risk subtypes (HPV-16, 18, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, 68, 73, 83 and MM4) and 5 low-risk subtypes (HPV-6, 11, 42, 43 and 44) and fixed them to the special membrane to make a DNA chip. A total of 112 male urethral samples were collected with swabs and studied for the clinical value. Meanwhile the single subtypes of HPV positive were sequenced and the standard samples detected for their sensitivity.</p><p><b>RESULTS</b>Of the total number, 25 samples were found to be HPV positive, 13 single HPV infection and 12 multiple infection. Nine HPV gene subtypes were noted in the samples: 6, 11, 16, 18, 33, 35, 43, 56 and 73, with sensitivity up to 10 copies of HPV DNA.</p><p><b>CONCLUSION</b>Human papillomavirus genotyping by the membrane DNA chip is applicable to the diagnosis of male HPV infection as well as to the related epidemic and etiological investigation.</p>

Roles of CD8+ CD28- T regulatory cells in acute infectious mononucleosis in children / 中华儿科杂志

<p><b>OBJECTIVE</b>To explore the role of CD(8)(+)CD(28)(-) T regulatory cells (Tr) in the immunological pathogenesis of acute infection with Epstein-Barr virus in children.</p><p><b>METHODS</b>The present study enrolled 25 children with infectious mononucleosis (IM) and 25 age-matched healthy children. Flow cytometric analysis was performed to detect the percentage of CD(3)(+), CD(3)(+)CD(4)(+), CD(3)(+)CD(8)(+), CD(8)(+)CD(28)(+) by determining the ratio of positive cells in lymphocytes. Reverse transcriptase-polymerase chain reaction (RT-PCR) and real-time PCR were used to analyze IL-6, IL-10, IFN-gamma expression in CD(8)(+)CD(28)(-) Tr cells and ILT-3, ILT-4 expression in monocytes/macrophages.</p><p><b>RESULTS</b>The proportions of CD(8)(+)CD(28)(-)T cells in children with acute-phase IM was significantly higher than those in the controls (P < 0.01). The expression level of IL-6, IL-10, IFN-gamma, ILT-3, ILT-4 mRNA significantly increased compared to those of the controls (P < 0.01).</p><p><b>CONCLUSION</b>The CD(28) expressed on CD(8)(+) T cells in vivo is gradually lost with age and CD(8)(+)CD(28)(-) cells increase up 50% to adult. EBV can directly infect B cells, trigger CD(8)(+) CTL response and destroy the target cells to cause serious immunopathological lesion. Therefore we speculate that the expansion of CD(8)(+)CD(28)(-) Tr cells in children with IM may be an adaptive immune response to avoid serious inflammation and autoimmune reactions.</p>

Relationship between Imbalance of Suppressors of Cytokine Signaling 1/Suppressors of Cytokine Signaling 3 and Abnormal Activation of Monocytes/Macrophages in Children with Asthma / 实用儿科临床杂志

Objective To investigate the relationship between the imbalance of suppressors of cytokine signaling 1(SOCS1)/SOCS3 and abnormal activation of monocytes/ macrophages in children with acute-phase asthma, and explore the molecular mechanism of chronic inflammatory process on airway.Methods The present study enrolled 20 asthmatic children and 20 age-matched normal children. Dual-color flow cytometric analysis was performal to detect the percentage of CD_ 80 、CD_ 86 expressing on CD_ 14 ~+ cell. Reverse transcriptase-polymerase chain reaction and real-time PCR were used to analyze SOCS1,SOCS3 expression in monocytes/ macrophages.Results The proportions of CD_ 80 ,CD_ 86 expressed on CD_ 14 ~+ cell in children with asthma were significantly higher than those in control subjects(CD_ 80 :7.0% vs 1.70%),(CD_ 86 :11.37% vs 2.03%),all P

Construction of a novel Schistosoma japonicum DNA vaccine pBK-Sj14-3-3 and studies on its immunoprotection in mice / 中华预防医学杂志

<p><b>OBJECTIVE</b>To prepare Sj14-3-3 DNA vaccine and observe its immunoprotection against Schistosoma japonicum in mice.</p><p><b>METHODS</b>The Sj14-3-3 gene was amplified by reverse transcription-polymerase chain reaction (RT-PCR) and subcloned into eukaryotic expression vector pBK. The recombinant plasmid pBK-Sj14-3-3 was extracted, purified and inoculated into BALB/c mice by intramuscular injection. Mice were attacked by Schistosoma japonicum cercariae and then killed. Adult worm and egg were counted, respectively. Diameter of the egg granulomas in the liver of infected mice was measured.</p><p><b>RESULTS</b>Electrophoresis on 1% agarose gel showed that the product of RT-PCR and the inserted fragment of recombinant plasmid digested with EcoR I and Xho I had the same size, about 765 bp, confirming the latter was the 14-3-3 encoding gene by nucleotide sequencing. Adult worm load declined by 27%, average egg load of per gram (EPG) of the liver tissues by 79%, average egg production per couple of adult worm (EPWP) by 51%, and mean diameter of egg granulomas by 29% in vaccinated mice.</p><p><b>CONCLUSION</b>The recombinant plasmid pBK-Sj14-3-3 was successfully constructed, which had some immunoprotection against Schistosoma japonicum in infected mice, indicating its potential to be vaccine candidate molecule of Schistosoma japonicum.</p>

Changs of B7 Family Costimulatory Molecules and Their Receptors Expression in Children with Asthma / 实用儿科临床杂志

Objective To explore the possible role of B7 family cell surface costimulatory molecules and their receptors in immunopathogenesis in children with asthma.Methods Forty children with acute exacerbation of asthma and 36 cases of age-matched healthy children were enrolled in the study.Interferon(IFN-?),IFN-? and lipopolysaccharide(LPS),phorbol 12-myristate 13-acetate and ionomycin or zero were used to stimulate peripheral blood mononuclear cells,flow cytometry was used to detect B7-1 and B7-2,B7H1,B7DC and B7H expression on CD14+ cells;CD4+ cells were separated by immune beads and stimulated by phorbol-12-myristate-13-acetate and ionomycin and fluorescence quantitative polymerase chain reaction was used to detect CD28,cytotoxic lymphocyte associated antigen-4(CTLA-4),programmed death-1(PD-1)and inducible costimulator(ICOS) mRNA expression.Results 1.The levels of B7-1 and B7-2 expression in children with asthma were higher than those of healthy control group [B7-1:(6.68?3.97)% vs(1.74?0.69)%;B7-2:(10.87?5.99)% vs(1.58?0.75)% Pa