AIM: To investigate the changes in intracellular potassium activity(a iK) and membrane potential(V_m) induced by low external sodium infusion (Low _o) and to detect the mechanisms involved and the relationship between a iK and V_m. METHODS: a iK and V_m were measured in infusion with different sodium concentrations using methods of convenient and ion selective microelectrodes in guinea pig ventricular myocardium. RESULTS: Low _o resulted in a decrease in a iK and an increase in V_m in a Na + concentration-dependent manner.At the same time,contraction and resting potential increased, and action potential duration decreased markedly,but action potential amplitude was not affected. A change of the pH from 7.4 to 7.0 in low _o solution reduced the decrease in a iK, but did not affect the increase in V_m. CONCLUSION: A better linear relationship appeared between the changes in a ik and _o or in V_m and _o,while during each low _o the change in both a iK or V_m may reach a new balance level.

ObjectiveTo assess the efficacy of herpes virus entry mediator (HVEM) gene modifled dendritic cells (DCs) in protecting against myosin induced myocarditis,and to investigate the involving mechanism.MethodsWe treated experimental autoimmune myocarditis (EAM) mice with myosin-pulsed DCs which were transfected with HVEM-expressing adenovirus (Ad-HVEM) or control vectors,then evaluated myocarditis,plasm cTn [ and autoantibody by histopathology,fluoroimmunoassay,and ELISA,respectively.ResultsWe found that DCs transfected with Ad-HVEM (DC-Ad-HVEM) could protect against EAM.Further study showed DC-Ad-HVEM could produce regulatory cytokine IL-10,and IL-10 promoted the production of a key regulatory T cell subset which is important in peripheral tolerance.The T cells mediated protection against EAM.ConclusionThis study suggest that myosin-DC-Ad-HVEM cell gene therapy is a safe and effective way for inhibiting the development of EAM,and the signal net mediated by HVEM plays different roles in different cells.

Objective: To explore the differences in the levels of 25 hydroxy vitamin D3 (vitamin D3) of the asthmatic children with different control levels, to analyze the relationships between the vitamin D3 level and the pulmonary function indexes, and to clarify the effects of vitamin D3 level on the asthma control and the pulmonary function. Methods: A total of 64 asthmatic patients who aged from 6 to 14 years old were selected as asttma group, and then according to the control level, asthma groups were divided into well-controlled group, partially controlled group and uncontrolled group. Eighteen healthy children who aged from 6 to 14 years old were selected as control group. The general materials of subjects in gwo groups were collected. The levels of serum vitamin D3 of the subjects in two groups were detected by Roche electrochemiluminescence, and the pulmonary function indexes (FEV1, FEV1 % pred, FEV1/FVC%, and FVC) were measured by spirometry. The levels serum of vitamin D3 of the subjects in asthma and control groups were analyzed by Wilcoxon rank sum test, the differences of vitamin D3 levels among different control level groups were analyzed by Kruskal-Wallis test, and the relationships between the vitamin D3 level and the pulmonary function indexes were analyzed by Spearman correlation analysis. Results: The serum vitamin D3 level of the children in asthma group was significantly lower than that in control group (P< 0. 05). The level of serum vitamin D3 of the children in well-controlled group was higher than that in uncontrolled group (P<0. 05), but the level of serum vitamin D3 of the children in partially controlled group had no differences neither with well-controlled group nor with uncontrolled group (P>0, 05). The percentage of children with vitamin D3 deficiency in uncontrolled group was significantly higher than that in well-controlled group (P<0. 05). The vitamin D3 level was positively correlated with FEV1 and FEVl/FVC% (r=0. 651, P<0. 01; r= 0. 642, P<0. 01). Conclusion: The level of serum vitamin D3 of the asthmatic children affects the control level of asthma and pulmonary function, so vitamin D3 inefficiency or deficiency should be paid attention in the treatment of asthma. Vitamin D3 supplementation is expected to be an important means of adjuvant treatment for asthma.

Objective To assess the sequence variations in preS/S regions of occult hepatitis B virus (OHB) and their relationship to severe chronic hepatic injury. MethodsWe collected samples from HBsAg negative patients, and evaluated their HBV-DNA by nest-PCR. HBV-DNA positive samples were used for analysis of preS/S region by PCR sequencing. Results Sixty-nine cases with HBV-DNA were identified in 468 cases without HBsAg. The positive percents were 16％, 8.7％, 36.4％, 18.3％ and 0％in group of only HBcAb positive, only HBeAb positive, only HBeAg positive, both HBcAb and HBsAb positive and all indexes negative, respectively. The level of HBV-DNA of OHB was significant lower than that in HBsAg positive patients. Compared with HBsAg positive controls, preS/S deletion, M1I and Q2K in preS2 region, Q129N/R/P,G185R and S210R in S region were more common in OHB. Moreover, M1I and Q2K in preS2 region, G185R and S210R in S region in OHB with severe chronic hepatic injury were more common that those in OHB without severe chronic hepatic injury. Compared with HBsAg positive patients with severe chronic hepatic injury, the level of HBV-DNA was lower, while the frequency of M1I and Q2K mutation in preS2 region, G185R and S210R in S region were more common in OHB patients with severe chronic hepatic injury. ConclusionThe virological factors were different between OHB and HBsAg positive patients. The M1I and Q2K in preS2 region, G185R and S210R in S region might be useful for prognosis evaluation of OHB patients.

Objective:To prepare gACE polyclonal antibody for functional study of gACE.Methods:According to the bioinformatics analysis and prediction of the possible conformational structure,hydrophobicity and antigenicity of gACE and the principal for antibody production,a partial peptide with 18-amino acid residues of gACE was synthesized after homology search.The synthesized peptide was then used to immunize after coupling with KLH.The properties of anti-gACE were analyzed by ELISA,Western blot and immunohistochemistry.Results:The antigenicity was repredicted by bioinformatics analysis.The polyclonal antibody against gACE was successfully obtained and its specificity and sensitivity we conformed by ELISA,Western blot and immunohistochemistry.Conclusion:By the bioinformatics analysis and prediction,the hydrophilicity and antigenicity of gACE are analyzed.The antibody of gACE is successfully obtained.

OBJECTIVE To investigate the possible effects of meteorological and environmental factors on IL-4 gene specific DNA methylation levels in CD4+T cells of children with AR.METHODS Thirty five pediatric AR patients(6-12 years) followed up for one year from the Department of Otolaryngology, Shanghai Children Medical Center from Jan, 1, 2012 to Dec. 31, 2012 were included in this study. Data on daily particulate matter of diameter smaller than 10 micrometer(PM10) and particulate matter of diameter smaller than 2.5 micrometer(PM2.5) was available as average values derived from the data of 6 state-controlled monitoring stations distributed across Pudong District, Shanghai. We quantified IL-4(interleukin-4) gene specific DNA methylation levels in CD4+T cells from 35 patients with AR and 30 healthy controls. mRNA levels of IL-4 gene were measured by real-time reverse transcriptase-PCR. Methods of personal exposure assessment of PM2.5 and PM10 were measured.RESULTS Compared with controls, IL-4 promoter region was hypomethylated in AR CD4+T cells(P=0.038). Of all observed CpG sites in IL-4 promoter region, there was significant differences in CpG-48, CpG+54(P=0.041, 0.032). IL-4 mRNA expression was significantly increased in CD4+T cells(P=0.039). The level of IL-4 mRNA expression was negatively correlated to the mean level of methylation in IL-4 promoter region. After adjusting, level of PM10 exposure was negatively correlated with level of methylation in IL-4 promoter region(r2=0.419,β=-0.470,SD=0.781,P=0.045). CONCLUSION Level of methylation in IL-4 promoter region may be affected by PM10 exposure.

[Objective]To investigate the effect of Baoyuan Jiedu Decoction(BJD)on serum lipids and white adipose tissue browning in cancer cachexia mice.[Methods]The specific pathogen free C57BL/6 mice were divided into normal group,model group,BJD group and megestrol acetate(MA)group.After 21 days of intervention,the changes of body weight,food intake,water consumption and tumor volume of the mice were observed,multidimensional mass spectrometry-based shotgun lipidomics(MDMS-SL)was used to determine the content of serum lipid of mice,white adipose tissue morphology and lipid droplet area were detected by hematoxylin-eosin(HE)staining,the expressions of white adipose tissue browning related genes were detected by Real-time quantitative polymerase chain reaction(Real-time PCR);and the protein expression of uncoupling protein 1(UCP1)was detected by Western blot and immunohistochemistry(IHC)staining.[Results]Compared with model group,the mice in BJD group were generally in good condition,and their food intake,water consumption and weight were increased significantly(P＜0.05),and the volumes of tumors were significantly suppressed(P＜0.05).Compared with normal group,there were 61 kinds of abnormal lipids in the serum of model group,while 30 kinds of lipids were influenced by BJD treatment(P＜0.05).Compared with model group,BJD alleviated the mass loss and lipid droplets(P＜0.05),inhibited the mRNA expression of UCP1,Cidea,Prdm16(P＜0.05)and the protein expression of UCP1(P＜0.05)in epididymal white adipose tissue(eWAT)and inguinal white adipose tissue(iWAT)of cancer cachexia mice.[Conclusion]BJD can inhibit weight loss,relieve the disorder of serum lipid,and inhibit the white adipose tissue browning of iWAT and eWAT of cancer cachexia mice.

Objective:To evaluate the effect of lung recruitment maneuvers combined with individualized positive end-expiratory pressure(PEEP) on the degree of postoperative atelectasis in elderly patients undergoing laparoscopic surgery.Methods:One hundred and forty-three elderly patients, aged ≥65 yr, with body mass index of 18.5-30.0 kg/m 2, scheduled for elective laparoscopic surgery, were assigned to either individualized PEEP combined with recruitment maneuvers (group Ⅱ) or fixed PEEP (group Ⅰ) using a random number table method. PEEP was maintained at 6 cmH 2O starting from the beginning of procedure until the end of the procedure in group I. Individualized PEEP titration was performed after induction of anesthesia in group Ⅱ. The primary outcome measure was the 12-zone lung ultrasound score at 15 min after tracheal extubation. Other outcome measures were the occurrence of postoperative pulmonary complications within 7 days after surgery, Quality of Recovery-15 scale score on 3rd day after surgery, rate of unplanned admission to intensive care units, length of hospital stay, incidence of intraoperative hypoxemia, usage rate of intraoperative vasoactive drugs, and incidence of postoperative hypotension. Results:Compared with group Ⅰ, the lung ultrasound score, driving pressure and postoperative pulmonary complications were significantly decreased, the dynamic lung compliance was increased ( P<0.05 or 0.01), and no significant changes were found in the other parameters in group Ⅱ ( P>0.05). Conclusions:Individualized PEEP combined with recruitment maneuvers can reduce the degree of postoperative atelectasis in elderly patients undergoing laparoscopic surgery.

Objective To investigate the phenotypic and functional properties of in vitro-induced He-lios+regulatory T cells(Helios+iTreg) and to analyze the differences between Helios+iTreg and Helios+thymic-derived natural Treg (Helios+nTreg). Methods CD4+CD25- effector T cells (CD4+CD25- Teff) that were separated from healthy subjects were cultured with anti-CD3 and anti-CD28 antibodies and stimulated with IL-2 and TGF-β to induce the generation of Helios+iTreg. Flow cytometry and real-time PCR were respectively used to analyze the differences in phenotype and CpG methylation in Treg-specific demethylated region (TSDR) be-tween Helios+iTreg and Helios+nTreg derived from the same donor. Results CD45RA was highly expressed on Helios+iTreg, but not on Helios+nTreg. Additionally, Helios+iTreg expressed significantly higher levels of CD127,ICOS and PD-1,but lower levels of CXCR3,CCR4 and CD25 than Helios+nTreg did. IFN-γ and IL-2 that expressed by Helios+iTreg were more than those by Helios+nTreg. It was also found that the level of CpG methylation in TSDR was much higher in Helios+iTreg than in Helios+nTreg. Conclusion Both nTreg and iTreg expressed Helios,but the phenotypic and functional properties of the two Treg subsets were different. It was suggested that Helios+iTreg and Helios+nTreg might play different roles in the immune system.

Objective To analyze changes in the percentages of circulating follicular helper T (Tfh) cells and Tfh subsets in patients with systemic lupus erythematosus (SLE) for better understanding their relationships with SLE , and to investigate effects of steroids on circulating Tfh cells .Methods Pe-ripheral blood mononuclear cells (PBMCs) from 27 patients with SLE (including 10 inactive patients and 17 active patients ) and 21 sex-and age-matched healthy donors were analyzed by flow cytometry to detect the percentage of CD4+CD45RA-CXCR5+Tfh-like cells.Disease activity and the concentration of anti-double-stranded DNA ( anti-dsDNA) antibody were evaluated by SLEDAI score ( SLE disease activity index ) and ELISA, respectively.PBMCs from healthy donors were treated with or without prednisone to evaluate its effects on circulating Tfh cells .Twelve patients with SLE were treated with high-dose steriods ( 200-500 mg/d, 2-3 d) and the percentages of circulating Tfh cells and Tfh subsets in them were analyzed before and after treatment .Results No significant difference in the percentage of circulating Tfh cells was ob-served between patients with SLE and healthy donors (P>0.05), but the percentage of Tfh17 cells in pa-tients with SLE was significantly higher than that in healthy donors (P<0.05).Compared with patients with inactive SLE and healthy donors , patients with active SLE had a lower percentage of Tfh 1 cells (P<0.05).Moreover, the percentage of Tfh1 cells was negatively correlated with SLEDAI score (r=-0.44, P<0.05). The percentage of Tfh2 cells in anti-dsDNA antibody-positive group was significantly higher than that in anti-dsDNA antibody-negative group (P<0.05).In vitro treatment of PBMCs from healthy donors with predni-sone could significantly down-regulate the percentage of circulating Tfh (P<0.01), Tfh1 (P<0.05) and Tfh2 cells (P<0.01), and up-regulate the percentage of Tfh17 cells (P<0.01).In vivo treatment of pa-tients with SLE with steriods could significantly down-regulate the percentage of circulating Tfh (P<0.01), Tfh1 (P<0.05) and Tfh2 cells (P<0.01) and up-regulate the percentage of Tfh17 cells (P<0.01).Con-clusion Abnormal distribution of Tfh subsets is correlated with SLE disease activity and anti -dsDNA anti-body .Steroids in the treatment of SLE could affect the percentage of circulating Tfh cells and the distribution of Tfh subsets .