Objective To investigate the effects of reactive oxygen species (ROS) inhibition on the down-regulation of adiponectin (ADPN) in mouse 3T3-L1 adipocytes by advanced glycation end-products (AGEs).Methods AGEs were prepared for incubating with cell.3T3-L1 preadipocytes were cultured in vitro and differentiated into mature adipocytes.Cell differentiation and lipid accumulation were determined by oil red O staining.After being intervened with AGEs,2',7'-dichlorofluorescein diacetate (DCFH-DA) was used as a reactive oxygen species (ROS) capture agent and the fluorescent intensity of 2',7 '-dichlorofluorescein (DCF) was detected by flow cytometry.Adiponectin expression under AGEs in 3T3-L1 adipocytes pretreated with N-acetyl-L-cysteine(NAC) or not was detected by real-time fluorescent PCR and ELISA.Results The level of ROS in 3T3-L1 adipocytes treated with AGEs was increased.mRNA and protein of ADPN were down-regulated.After inhibition with ROS,mRNA and protein expressions of ADPN injured by AGEs were ameliorated.Conclusion Exposure of 3T3-L1 adipocytes to AGEs induces oxidative stress in vitro,which decreases the expression of ADPN,and causes functional impairment of adipose cells and insulin resistance.

Objective To prepare the chitosan-potD nanoparticles and to evaluate its protective efficacy against pneumococcal nasopharyngeal colonization. Methods potD gene was amplificated from pneumococcal genome and was inserted into pVAX1 expression vectors to construct pVAX1-potD recombinant plasmid which was then transfected into 293T cell using LipofectAMINE 2000 to analyze transient potD gene expression in vitro by RT-PCR and Western blot. Chitosan-potD nanoparticles were freshly prepared by coacervation methods at each time and the characterizations of the nanoparticles were then evaluated. BALB/c mice were immunized with chitosan-potD, naked potD DNA or pVAX1 for 4 times at two-week intervals. Anti-PotD IgG, IgG1 and IgG2a levels in serum and IgA levels in nasal washes, bronchoalveolar lavage fluids (BALF) and middle ear lavages(MEL) were detected by indirect enzyme-linked immunosorbent assay (ELISA). IL-17A, IL-4 and IFN-γ levels in splenocytes were determined by double sandwich ELISA. Mice were intrannsally challenged with Streptococcus pneumoniae ATCC6303, and Pneumococci were recovered from the nasopharyngeal niche at the fifth day after challenge. Results potD gene was successfully amplificated by PCR and the sequence was confimed to be consistent with that in the Genbank. The pVAX1-potD recombinant plasmid was successfully constructed and was expressed in eukaryocytes in vitro. The mean size and zeta potential of chitosan-potD nanoparticles was 430 nm and + 20.5 mv, respectively. Chitosan-potD nanoparticles were not digested by DNase Ⅰ , while naked potD DNA was completely digested. The levels of antibodies inculding IgG, IgG1, IgG2a, IgA and cytokines including IL-17A, IL-4 and IFN-γ were significantly higher in mice immunized with chitosan-potD nanoparticles than mice with naked potD or pVAX1 ( P <0.05) only. More importantly, much less Pneumococci were recovered from mice immunized with chitosan-potD nanoparticles than the other groups(P <0.05). Conclusion Chitosan-potD nanoparticles significantly enhanced the immunogenicity and protection efficacy of DNA vaccines by intranasal immunization and could be used as a potential mucosal vaccine to prevent pneumococcal infection.

The abdominal visceral fat content in obese SD rats induced by high fat diet for 10 weeks was significantly higher than that in control group [(26±6 vs 13±3)g,P<0.01] ,along with increased CRP mRNA expression in abdominal visceral fat (0.901±0.085 vs O. 402±0.036, P<0.01). As compared with normal control group, in the high fat group the concentrations of CRP in portal vein [(743.8±95.8 vs 558.3 ±118.3) mg/L, P<0.01] and peripheral vein[(596.3±38.9 vs 485.8±30.2) mg/L,P<0. 05] were higher. The concentration of CRP in portal vein was significantly higher than that in peripheral vein in high fat diet group(P<0.01) ,but this was not evident in control group. These results suggest that the increased CRP expression in visceral adipose tissue may partially account for the elevation of serum CRP in obesity.

Objective To discuss the feasibility and value of indocyanine green combined with methylene blue in sentinel lymph node biophy (SLNB) for patients with breast cancer.Methods 196 breast cancer patients who received surgery were studied.SLNB was performed by indocyanine green combined with methylene blue .Frozen section was performed,followed by conventional histopathology .The evaluation criterion of Louisvill University on SLNB was adopted to judge the detection rate ,accuracy rate,sensitivity,false negative rate.Results Among the 196 cases,196 cases were detected with SLN ( detection rate was 100.0%).SLN was all stained in 196 cases. Furthermore,56 cases had ALND metastasis,140 cases had no metastasis,and the sensitivity was 91.8%(56/61). The SLN and ALN fully complied with pathological examination in 191 cases,and the accuracy rate was 97.5%(191/196).Five cases were false negative,and the false negative rate was 8.2%(5 /61).No case was false positive.The total SLNs was 705,and average number of detected SLNs was 3.59 pieces(2-6 pieces).The average ALN was 17.66 pieces(12-26 pieces).Conclusion With the advantages of real-time visual,precise positioning,no radio-active pollution and reduce the operation difficulty ,indocyanine green joint methylene blue as lymph tracer shows high detection rate,accuracy when it is used for SLNB in patients with breast cancer ,it is worthy of clinical application.

Objective To explore the correlation of the timing of enteral nutrition (EN) and the prognosis of the patients with septic shock.Methods From December 2016 to August 2017,fifty-five patients with septic shock who received treatment in the ICU of Tongji University School of Medicine were selected in the study and randomly divided into two groups,28 cases in the early enteral nutrition (EEN) group and 27 cases in the delayed enteral nutrition (DEN) group.All the patients was treated with EN before and after 48 h at admission respectively.After 1 week of treatment,the nutrition indicators,clinical efficacy,complications and prognosis of the two groups were compared.Results The levels of ALB,PA and HB in EEN group after treatment were significantly increased,compared with those before treatment,and were superior to those in DEN group ((38.4±4.5) g/L vs.(34.6±3.8) g/L,(207.8±41.4) mg/Lvs.(180.6±47.6) mg/L,(119.2 ±18.3) g/Lvs.(110.7±14.2) g/L;t =3.39,2.26,1.99,P＜0.05).Compared with DEN group,the length of ICU stay,CRRT time,mechanical ventilation time of EEN group were significantly shortened ((6.4± 1.7) d vs.(9.8±2.4)d,(4.7±1.2) d vs.(7.9±2.5)d,(3.6±1.2)d vs.(6.4±1.8) d;t=6.16,6.32,7.09,P＜0.05).After treatment,the SOFA score,APACHE Ⅱ score and qSOFA score were significantly improved in both groups,and the improvement in EEN group was significantly better than that in DEN group((3.8±0.9)points vs.(5.1 ± 1.2) points,(12.2 ± 2.4) points vs.(15.1 ± 3.7) points,(87.4 ± 14.7) points vs.(77.4 ± 13.2) points),(t=4.53,3.46,4.31,P＜0.05).Conclusion The implementation of EEN in patients with septic shock can improve the nutritional status of the patients,alleviate the severity of the disease,reduce the incidence of complications and improve the prognosis.