Objective To construct a prokaryotic expression system of gingipain R(Rgp) genes to identify the correlation between Porphyromonas gingivalis(Pg) gingipain R and chronic periodontitis.Methods The routing Phenol-chloroform method was applied to prepare genomic DNA of Pg ATCC33277.Oligonucleotide primers were designed to amplify the RgpAcat and RgpB genes according to the published corresponding nucleotide sequences.Prokaryotic expression system was established,and Ni-NTA affinity chroma-tography was applied to extract and purify the recombinant proteins.rRgpAcat and rRgpB were applied to human monocyte THP-1 cell strain.Flow-cytometric analyses were performed to detect the expression of CD-14 on the cell surface.Enzyme-linked immunosorbent assay( ELISA ) was used to detect the inhibition of lipopolysaccharides-induced IL-1β production from human monocytes by rRgpAcat or rRgpB.Results In comparison with the corresponding sequences from GeneBank,homologies of the nucleotide sequences of the cloned RgpAcat and RgpB genes were ＞ 97％.rRgpAcat and rRgpB output of the constructed prokaryotie expression system was approximate 50％ of the total bacterial proteins.The intensity of fluorescence of expression of CD-14 by THP-1 cells was 68.97.After rRgpAcat or rRgpB treatment with THP-1 cells for 0.5 h,it reduced to 45.30,46.47 (P ＜ 0.05 ),and the level of IL-1 β was also decreased (P ＜0.01 ).Conclusions Prokaryotic expression systems of RgpAcat and RgpB genes were successfully established in this study.rRgpAcat and rRgpB could cleave monocyte CD-14,block the THP-1 cells secreting cytokines,and as a consequence sustain chronic inflammation.

The aim of this study is to assess the status of treatment of chronic prostatitis (CP) in Chinese men. A population-based cross-sectional survey was performed, in which 15 000 men aged between 15 and 60 years were randomly selected to receive a questionnaire designed to assess National Institutes of Health Chronic Prostatitis Symptoms Index (NIH-CPSI) status, therapeutic efficacy and 28 other items. A total of 12 743 men (84.95%) completed the questionnaire, of whom 1 071 (8.4%) were identified as having prostatitis-like symptoms and 517 (4.5%) were diagnosed with CP according to NIH-CPSI criteria and prostatitis-like symptomatology. Of the CP patients, 372 (65%) underwent long-term routine treatment 12 times per year. Additionally, 217 (72.8%) patients received antibiotic therapy and 215 (79.3%) men showed therapeutic effects. The treatment cost USD 1 151 (8 059 yuan) per person per year on average. Most CP patients received routine treatment, in most cases with antibiotics. Treatment was costly and most CP patients were not satisfied with its effectiveness. Antibacterial treatment might have been effective primarily in patients with bacterial disease.
Adolescent ; Adult ; Anti-Bacterial Agents ; therapeutic use ; Bacterial Infections ; complications ; drug therapy ; epidemiology ; China ; epidemiology ; Chronic Disease ; Cross-Sectional Studies ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Male ; Middle Aged ; Prevalence ; Prostatitis ; drug therapy ; epidemiology ; microbiology ; Surveys and Questionnaires ; Treatment Outcome ; Young Adult

The purposes of this study was to determine the effects of recombinant human interleukin-10 (rhIL-10) on proliferation of vascular smooth muscle cells (VSMCs) stimulated by advanced glycation end products (AGE) and neointima hyperplasia after rat carotid arterial injury. Rat aortic VSMCs were cultured and treated with rhIL-10 or AGE respectively, and then co-treated with rhIL-10 and AGE. Proliferation of VSMCs was quantified by colormetric assay. Cell cycle analysis was performed by flow cytomertry. Sprague-Dawley rats were treated with recombinant human IL-10 (rhIL-10) for 3 d after carotid arteries injury. The ratio of neointima to media area at the site of arterial injury was measured 28 d after balloon injury. The p44/42 MAPK activity was evaluated by the immunoblotting technique using anti-p44/42 phospho-MAPK antibody. Compared to control, AGE stimulated VSMCs proliferation. rhIL-10 alone had no effect on VSMCs growth. With AGE stimulation, rhIL-10, at dose as low as 10 ng/ml, inhibited VSMCs growth (P<0.05). The cell number in G(0)/G(1) phase of AGE and rhIL-10 co-treatment group was higher than that of AGE treatment alone (P<0.01) by flow cytometry analysis. Compared with the control group of neointima hyperplasia in rats, the ratio of neointima to media area of recombinant human IL-10 group was reduced by 45% (P<0.01). The p44/42 MAPK activity was significantly enhanced by AGE. The AGE effects were opposed by rhIL-10. The anti-inflammatory cytokine rhIL-10 inhibits AGE-induced VSMCs proliferation. Recombinant human IL-10 also inhibited neointima hyperplasia after carotid artery injury in rats. The results suggest the possibility that recombinant human IL-10, as a potential therapeutic approach, prevents neointimal hyperplasia.
Animals ; Aorta, Thoracic ; cytology ; Atherosclerosis ; physiopathology ; Carotid Artery Injuries ; pathology ; physiopathology ; Carotid Intima-Media Thickness ; Cell Proliferation ; drug effects ; Cells, Cultured ; Glycation End Products, Advanced ; antagonists & inhibitors ; pharmacology ; Hyperplasia ; prevention & control ; Interleukin-10 ; pharmacology ; Male ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; drug effects ; Neointima ; drug therapy ; prevention & control ; Rats ; Rats, Sprague-Dawley ; Recombinant Proteins ; pharmacology ; Tunica Intima ; pathology