Bone Density Conservation Agents ; Dental Implants ; Diphosphonates ; Humans

Objective:To establish a double antibody sandwich ELISA assay for detection of human IL-37 in serum.Methods: Mouse anti-human IL-37 monoclonal antibody was used as capturing antibody,rabbit anti-human IL-37 polyclonal antibodies served as detection antibody,HRP labeled goat anti-rabbit IgG employed as second antibody and recombinant human IL-37 protein used as reference standard for the establishment of a doubleantibody sandwich ELISA.The working conditions were optimized,such as sensitivity,linear range,reproducibility and evaluated the serum IL-37 in patients with Dengue fever.Results: The sensitivity of the established ELISA method was 1.465 μg/L approximately.Likewise,the linearity range of this method was about (1.465-46.875) μg/L.Further,the co efficient of variation (CV) of inter-batch and intra-batch in this study were 6.6％ and 11.7％,respectively.Notably,this method could be used in the detection of IL-37 in serum of the patients with Dengue fever,showing that the level of IL-37 in Dengue fever patients was much higher than that in healthy controls.Conclusion: The double antibody sandwich ELISA assay for the detection of human IL-37 was successfully established,which can be apply to detect of human IL-37 in clinical samples.

Objective To investigate the antimicrobial resistance ,molecular phenotypes ,virulence gene profiles of Salmonella A gona (S .A gona) isolated from patients with acute diarrhea ,and to better understand its epidemic trend ,prevention and treatment .Methods Clinical data and stool samples of patients with acute diarrhea during April to October in 2013 and 2014 from the Second Hospital of Tianjin Medical University were collected .Enrichment culture ,biochemical identification and serotyping analysis were used to isolate and identify S .A gona strains .The isolated strains were further analyzed with antibiotics susceptibility test ,pulsed field gel electrophoresis (PFGE) ,multiple locus sequence typing (MLST ) , Quinolone resistance determining region (QRDR) .Plasmid-mediated quinolone resistance (PMQR) and β-lactamases genes (TEM ,SHV ,OXA ,and CTX-M) were amplified by polymerase chain reaction (PCR) and sequencing .The representative genes carried by Salmonella pathogenicity islands (SPI) 1 — 6 ,9 — 12 and virulence plasmids were amplified by PCR .And the clinical characteristics of S .Agona infection were analyzed .Results Among 119 non-repetitive (non-typhoidal salmonella ,NTS) isolates during the two years ,eight isolates (6 .7％ ) of S .A gona were identified . The resistance rate of S .A gona strains to streptomycin was 100 .0％ , those to ampicillin and gentamicin were 62 .5％ ,to levofloxacin ,ciprofloxacin and nalicixic acid were 25 .0％ ,to chloramphenicol ,amoxillin/clavulanic acid and piperacillin tazobactam were 12 .5％ .The strains were susceptible to other drugs .All 8 isolates had the identical ST13 genotype .PFGE showed 5 clones ,and 4 out of 5 isolates had the exact same patterns of PFGE and drug susceptibility .Two (fluoroquinolones ,FQ) resistant strains carried gyrA mutation leading to amino acid substitutions at position 87 in GyrA ,and no PMQR genes was detected ,while one of which was sensitive to ciprofloxacin by K-B method .All five ampicillin-resistant isolates were positive for TEM-1b gene and one isolate of them was resistant to β-lactam/β-lactamase inhibitor complex .The representative genes carried by SPI 1 — 6 , 9 ,11 ,12 (hilA ,sseL ,mgtC ,siiE ,sopB ,pagN ,bapA ,pagC and sspH2) were 100 .0％ positive ,while the genes carried by SPI10 (sef A ) virulence plasmids (spvB , prot6E) were negative . Two patients with FQ resistant strains infection were clinically diagnosed with bacillary dysentery ,and the remaining six cases with FQ susceptible strains infection were clinically diagnosed with acute gastroenteritis .Conclusions FQs-resistant and multi-drug resistant S .A gonaisolates have emerged in clinical settings .These isolates carry a variety of virulence genes .Resistance to FQ of S .Agonamay cause more severe illness .ST13 might be the dominant genotype of S . A gona in China ,and we should try to prevent the infection outbreak of S .A gona .

Objective:To explore whether the electroacupunture stimulation (ES) at acupoint Zusanli (ST36) can inhibit the bone loss caused by Staphylococcus aureus (SA) infection and its mechanism in a model of SA osteomyelitis.Methods:Twelve male C57 BL/6 mice aged 10 to 12 weeks were randomly divided into 2 even groups ( n=6) for SA infection + ES or SA infection only. After ES at ST36 was conducted for 4 weeks in the model of SA osteomyelitis, samples were harvested from the femora and tibiae. Micro-CT reconstruction was performed to detect trabecular bone volume fraction (BV/TV), trabecular number (Tb.N), trabecular thickness (Tb.Th), trabecular separation (Tb.Sp), connectivity density (Conn.Dn) to analyze changes in bone mass. Leptin receptor (LEPR) staining was performed to detect osteoblasts. Tartrate resistant acid phosphatase (TRAP) staining was used to detect the changes in osteoclasts. The changes in plasma inflammatory factors were detected by enzyme-linked immunosorbent assay (ELISA). Results:Micro-CT results showed that the BV/TV, Tb.N, Tb.Th, and Conn.Dn in the cancellous bone in the target areas in the SA + ES group were all higher than those in the SA group, LEPR immunofluorescence results indicated that the number of osteogenic precursor cells in the ES group was larger than that in the SA group, and serum ELISA indicated a decrease in inflammatory factors in the blood in the SA+ES group compared with the SA group. There were significant differences in the comparisons above ( P<0.05). There was no significant difference in the number of osteoclasts on the surface of trabecular bone between the 2 groups in TRAP staining. Conclusion:ES may slow down infectious bone destruction by inhibiting the inflammatory response induced by SA infection and by inducing aggregation and differentiation of mesenchymal stem cells into trabecular bone.