Objective To investigate the effect of rapamycin(RPM)on hepatic interleukin-10(IL-10)gene and acute liver injury in rats with postburn Staphylococcus aureus sepsis.Methods Fifty male Wistar rats were randomly divided into four groups:normal control group,scald control group,postburn sepsis group,and RPM treatment group.Tissue samples from liver and plasma were collected to determine IL-10 mRNA and protein expressions,and liver function parameters were also measured.Results Compared to postburn Staphylococcus aureus sepsis group,in RPM treatment group hepatic IL-10 mRNA expression and plasma IL-10 were significantly increased at 0.5 hour after RPM treatment(P

Objective To investigate the effect of the inhibitor SB203580 of p38 mitogen-activated protein kinase (MAPK) signal transduction pathway on biopterin (BH 4)/nitric oxide (NO) expression and elucidate the potential mechanism of MAPK in biopterin-mediated NO induction after endotoxic shock. Methods A total of 56 male Wistar rats were randomly divided into normal control group (n=8), endotoxic shock group (n=32) and SB203580 treatment group (n=16). After animals were sacrificed, tissue samples from the liver, the lungs as well as the kidneys were harvested to determine the expressions of guanosine triphosphate cyclohydrolase I (GTP-CHI) and inducible nitric oxide synthase (iNOS) mRNA and observe the changes of BH 4 and NO levels in blood and tissues. Results With endotoxin challenge, GTP-CHI mRNA expression and BH 4 levels were significantly elevated in various tissues and maintained at high levels up to 24 hours. Similarly, the iNOS mRNA expression and NO levels in the tissues significantly increased too, especially in the liver and the lungs. Treatment with SB203580 significantly down-regulated GTP-CHI mRNA expression in the liver, the lungs and the kidneys at 12, 24 and 2-12 hours, respectively (P

PEI Xue-yi has 60 years,experience in treating pediatric diseases with traditional Chinese medicine,especially obtains good effect in children's nephrapiathy.Based on the regulation and characteristics of children's zang-fu,yin-yang,blood-qi and children's nephrapiathy,he makes different therapeutic plans in different phases.In clinical treatment,he emphasizes the lung,the spleen and the kidney and combines syndrome differentiation with disease diagnosis.Eliminaing pathogens,excessive damp-heat is used in acute phase and strengthening healthy qi,securing the spleen and kidney is used in recovery phase.Children's nephrapiathy is treated by three phases as the edema,protein urine and rescovery stage.He adopts the methods of dispersing lung qi and invigorating spleen for diuresis,clearing heat-damp,nourishing spleen and kidney,nourishing yin for protecting lower jiao.

Objective: To prepare naringenin-loaded solid lipid nanoparticles (NRG-SLN) lyophilized powder, and investigate its physicochemical properties and release characteristics, then to investigate the pharmacokinetic characteristics in rats after pulmonary delivery. Methods: NRG-SLN were prepared by solvent emulsification-evaporation method, the formulation was optimized by orthogonal design, with encapsulation efficiency as reference, and the measurements of particle size, morphology, Zeta potential, the polydispersity index (PDI) and in vitro drug release behavior were performed. To screen the best lyoprotectants in appearance, color, and redispersibility as indexes the differential scanning calorimetry (DSC) was used to analyze its material phase of the drug in nanoparticles. The study on pulmonary pharmacokinetics in rats was carried out by pulmonary instillation. Results: The NRG-SLN assumed a spherical shape with an even distribution of diameter and particle size of (97.69±2.84) nm, the PDI was 0.207±0.010, Zeta potential was (-26.20±0.45) mV, entrapment efficiency was (81.09±1.37)%, and drug loading was (8.30±0.04)% (n=3). Mannitol (5%) was the best protective agent for lyophilized powder of NRG-SLNs. The characterization indicated that the drug to amorphous state dispersed in a lipid. In vitro dissolution experiments showed NRG-SLN compared with pure drugs had obviously sustained release. After pulmonary administration to rats, the pharmacokinetic parameters of NRG-SLN and solution were as follows: Cmax (163.00±23.05) and (269.00±35.34) ng/mL, AUC0-t (929.32±190.28) and (3 390.23±533.68) ng∙h/mL, t1/2 (5.13±0.23) and (18.93±7.90) h, MRT (7.19±0.44) and (23.29±9.27) h. Conclusion: The technique of preparing NRG-SLN by solvent emulsification-evaporation has small particle size, high entrapment efficiency, and good stability, and the process is simple. Compared with the naringenin solution, the SLN show the sustained-release characteristics and can significantly improve the bioavailability after pulmonary administration.

Objective To observe the expression law of cytokine signaling suppressors (SOCSs) mRNA in burn rats with Staphylococcus aureus sepsis and investigate their potential role in the pathogenesis of postburn sepsis. Methods Wistar rats were inflicted with 20% TBSA Ⅲ? scald, followed by Staphylococcus aureus challenge. Then, the expressions of SOCS1, SOCS2 and SOCS3 mRNA and interferon-? (IFN-?) levels in the liver and lungs were determined. Results With Staphylococcus aureus challenge after burn, IFN-? levels in the liver and lungs were significantly elevated and reached peak at the 0.5th and 6th hours, respectively (P

Objective To construct a model for persistent genital tract infection with Chlamydia trachomatis in mice with low serum iron. Methods Iron deficiency was induced in female BALB/c mice by lowiron diet. Mice with or without iron-deficency were inoculated intravaginally with direct instillation of 50 μl of bacterial suspension containing 3.4344 × 107 IFU/ml of C. trachomatis serovar E elementary body 1 week after intramuscular injection of progesterone. Moxifloxacin and ferrous sulfate, alone or in combination were given to inoculated mice with iron deficiency daily for 1 week. Subsequently, vaginal discharge was obtained from mice and observed under an immunofluorescence microscope, and tissue specimens were resected from the uterus,uterine tube and ovary of mice and subjected to pathological examination at various time points. Results Compared with normal-iron mice, iron-deficient mice were infected with C. trachomatis for a longer duration (more than 18 weeks). After 18-week infection with C. trachomatis, there was an obvious chronic inflammation and lymphocytic infiltration in tissues specimens from the uterus,uterine tube and ovary of mice. Immunofluorescent examination of vaginal discharge for C. trachomatis turned negative in 60% of iron-deficient mice treated with both moxifloxacin and ferrous sulfate, but not in any of those receiving moxifloxacin or ferrous sulfate alone. Conclusion As animal models show, low serum iron may contribute to the persistent genital tract infection with C. trachomatis.

Objective To investigate the immunological activity change of regulatory T cells (Treg) and discuss its significance in the outcomes of patients with multiple organ dysfunction syndrome (MODS) and severe burn. Methods A total of 106 patients with total burn surface area (TBSA) larger than 30% were included in the study and randomly divided into three groups according to the burn area: Group Ⅰ (TBSA of 30%-49%, n = 41), Group Ⅱ (TBSA of 50% -69%, n = 34) and Group Ⅲ (TBSA of 70%-99%, n = 31). According to the development of MODS, patients were divided into MODS group (n =21) and non-MODS group (n =85). The patients with MODS were further divided into non-survival group (n = 16) and survival group (n = 5) based on their outcomes. Healthy volunteers were served as normal control (n = 25). Peripheral blood samples were collected at days 1,3,7, 14 and 21 postburn. The immunomagnetic separation technique was applied to separate and purify CD4+ CD25+Tregs in peripheral blood, and phenotypes (CTLA-4) were analyzed by flow cytometry and the contents of interleukin-10 released in the supernatants were determined by ELISA. Results Expression of CTLA-4 and level of IL-10 were significantly increased in burn patients compared with normal control group, with statistical differences. The expression of CTLA-4 and level of IL-10 were significantly increased in patients with severe burns at all time points. The expression of CTLA-4 and level of IL-10 in MODS group were much higher than those in non-MODS group at days 3-21 postburn (P ＜ 0.01). Among the MODS patients, the expression of CTLA-4 and level of IL-10 in the survival group were obviously lower than those in the non-survival group at days 3-21 postburn (P ＜ 0.05 or P ＜ 0.01). Conclusions After severe burn injury, expressions of the markers on CD4 + CD25 + Treg surface and secretion of cytokines produced by CD4 + CD25 + Tregs show significant difference in patients with different born areas, MODS development and survival state. CD4 + CD25 + Treg may play an important role in the pathogenesis of immunoregulation, MODS and mortality of burn patients through secretion of inhibitory cytokines.

Objective To clone, express and purify Chlamydia trachomatis polymorphic membrane protein (Pmp G), and to identify its immunogenicity. Methods The Pmp G gene of C. trachomatis serotype E was amplified by PCR, cloned into prokaryotic expression vector PET30a (+). The positive recombinant was transformed into the bacterium E coli (BL-21), identified by enzyme digestion, PCR amplification and gene sequencing. Then, it was induced to express followed by the identification of expression product with SDS-PAGE and Western blotting. The purified protein was used to immunize BALB/C mice to test its immunogenicity. Results PCR produced a 1092 bp-sized DNA fragment, which had a sequence consistent with that of PmpG gene of C. trachomatis E type in the GenBank database. The molecular weight of expression product was 55 kD, which was proved to be the expected size, and Western Blotting confirmed it to be the specific protein. Moreover, special antibodies to PmpG were induced to be generated by mice immunized with the purified protein. Conclusions The constructed prokaryotic expression vector for PmpG is expressed successfully in E. coli, and the expression product shows immunogenicity.

Objective:To investigate the effects of Helicobacter pylori on NLRP3 inflammasomes activation in THP-1 ( human monocytic cell line) -derived macrophages and evaluate the role of ROS.Methods:H.pylori strain SS1 was co-cultured with the THP-1-derived macrophages at a multiplicity of infection (MOI) of 1∶100 based on trial results with different MOIs (ratios of THP-1 cells to bacteria ranging from 1∶25 to 1∶200).The co-culture supernatants and THP-1 cells were collected at various time points (3 h,6 h,12 h,24 h) and cytokine production was quantitated using ELISA analysis.The generation of intracellular ROS was detected by FCM,and the mRNA transcript levels of NLRP3 and caspase-1 were measured by Real-time PCR.Western blot was employed to analyze the expression of active caspase-1 subunit ( p10).Then we observed the inhibitory effects of NAC and siRNA specific for NLRP3 on the ex-pression of NLRP3 inflammasome-related components and the secretion of cytokines induced by H.pylori.Results:We found that H.pylori SS1 induced IL-1βand IL-18 production in human acute monocytic leukemia cell line THP-1 in a time-and dose-dependent manner.We further showed that H.pylori could induce the mRNA expressions of NLRP3 and caspase-1 in THP-1 cells.Moreover, release of IL-1βand IL-18 from H.pylori-infected THP-1 cells was suppressed by the ROS scavenger NAC,which was an agent known to inhibit NLRP3 inflammasome formation.NAC administration also resulted in a significant decrease in the level of H.pylori-induced caspase-1 protein expression in THP-1 cells.Additionally,secretion of IL-1βand IL-18 in response to H.pylori infection was remarkably reduced by NLRP3-siRNA.Conclusion:The induction of IL-1βand IL-18 secretion by H.pylori strain SS1 in THP-1 cells could be mediated through activation of NLRP3 inflammasome via ROS signaling pathway, which may be involved in the host innate immune defence and the pathogenesis of the bacteria.

Absract: Objective To report the result of annual monitoring and analysis of nasopharyngeal virus in children with respiratory tract infections in Nanxiang, Shanghai District. Methods Nasopharyngeal secretions were collected from 4389 children with acute respiratory tract infection in outpatient department from January 2007 to September 2013, 9 common respiratory viruses were analyzed by Multiplex RT-PCR, including inlfuenza virus (FLU), parainlfuenza virus (PIV), respiratory syncytical virus (RSV) , adenovirus (ADV), human bocavirus(HBOV), human coronavirus(Cov), enterovirus(EV), human metapneumovirus(HMPV), and rhinovirus(HRV). The same analysis was done in 123 asymptomatic children during the same period. Results The positive rate of detected respiratory viruses in children with respiratory tract infections in nasopharyngeal secretions were 34.8% (1526/4389), including FLU 10.3% (453/4389), RSV 7.3% (320/4389), PIV 6.2%(274/4389), ADV 3.3%(146/4389), HBOV 2.7%(118/4389), EV 2.5%(110/4389), Cov 2.4%(105/4389), HRV 1.6%(72/4389), HMPV 1.5%(67/4389);two and more combined respiratory viral infection were found in 273 cases (6.2%). The virus detection
rate between age groups was signiifcantly different (χ2=41.91, P<0.001). The school-age group had the lowest positive rate of 23.4%and the positive rates in other three groups were all higher than 35.0%. The infant group had the higher positive rate of RSV and HRV. FLU detection rate in school-age group was 13.6%. Respiratory viruses in children with asthmatic disease has high detection rate. RSV infection rate was the highest 14.8%(30/204) in the asthmatic disease group, followed by HBOV 13.8% (28/204). In nasopharyngeal secretions of 123 asymptomatic children, virus-positive detection rate of 6.5% (8/123), which showed signiifcant difference from that in respiratory virus infection group (χ2=42.60, P<0.001). Conclusions In seven consecutive years of testing, the inlfuenza virus and respiratory syncytial virus play an important role in children with respiratory tract infections in this region. The detection rate of virus showed difference between different age groups and a higher detection rate of RSV in infants with respiratory tract infections was observed. The overall detection rate of virus was decreased with the increase of age excluding the inlfuenza virus.