We investigated the roles of the crude antigen(CA) of Clonorchis sinensis and excretory secretory products (ESPs) in the polarization of Th1 and Th2 cells.Bone marrow-derived cells were generated from BALB/c mice and isolated into immature DCs;immature DCs were then treated with either CA (CA stimulated group),ESPs (ESPs stimulated group),LPS (positive control group) or PBS (negative control group) for 24 hours.Then the CD4+T cells were isolated from mouse spleen by using anti mouse-CD4 Microbeads,and further cocultured with stimulated DCs for another 72 hours.The purities of DCs and CD4+ T cells were evaluated by flow cytometry and the expressing levels of T-bet mRNA and GATA-3 mRNA were detected by real-time PCR.ELISA was used to detect the levels of IFN-γ and IL-4 cytokines in the supernatant.mRNA levels of T-bet and GATA-3 in the ESPs group were higher than those in PBS-stimulated group (P＜0.05).The concentrations of IFN-γ and IL-4 cytokines in the culture were increased in the ESPs group,compared with PBS stimulated group(P＜0.05).IFN-γ but not IL-4 was increased in CA group (P＜0.05).The results implied that CA might play a role in Th1 type immune response,and ESPs likely play roles in both Th1 and Th2 immune responses.

Background:Wheezing is common in early childhood and remains an important health concem.The aim of this study was to assess the lung function of wheezing infants and to investigate the relationship between lung function and respiratory outcome.Methods:Infants ＜2 years of age with acute lower respiratory tract infection (ALRTI) who had undergone lung function tests were included in the study.They were assigned to wheeze or no wheeze group based on physical examination.Infants without any respiratory diseases were enrolled as controls.Lung function was measured during the acute phase and 3 months after ALRTI.One-year follow-up for infants with ALRTI was achieved.Results:A total of 252 infants with ALRTI who had acceptable data regarding tidal breathing were included in the final analysis.Compared with the control and the no wheeze groups,infants in the wheeze group had significantly decreased time to peak tidal expiratory flow as a percentage of total expiratory time (TPTEF/TE) (20.1 ± 6.4％ vs.34.4 ± 6.2％ and 26.4 ± 8.3％,respectively,P ＜ 0.0001) and significantly increased peak tidal expiratory flow (PTEF) (90.7 ± 26.3 ml/s vs.79.3 ± 18.4 ml/s and 86.1 ± 28.0 ml/s,respectively,P ＜ 0.01),sReffand Reff.The infants in the wheeze group still had lower TPTEF/TE and volume to peak tidal expiratory flow as a percentage of total expiratory volume (VPTEF/VE) than the no wheeze infants 3 months after the ALRTI.Moreover,there was a significant inverse relationship between TPTEF/TE,VPTEF/VE,and the recurrence of wheezing and pneumonia.Conclusions:Impaired lung function was present in wheezing infants with ALRTI and the deficits persisted.In addition,the lower level of TPTEF/TE and VPTEF/VE was a risk factor for poor respiratory outcome.

Objective To study the effects of heat shock proetin 27(Hsp27)on the cardiac dysfunction induced by endotoxemia.Method All experiments were performed in the geriatric lab of the First Affiliated Hospital Of Nanjing Medical University,and in the Animal Model Center of Nanjing University.The genotyping of the transgenic mice with cardiac-specific overexpression of Hsp27(Hsp27 Tg)was assayed by PCR and the expression of Hsp27 was determined by western blot.Hsp27 Tg and its wild type littermates(WT)were intraperitoneally injected with LPS(10 mg/kg),and 24 hours later,cardiac function was measured by echocardiography(n=6/group).The accumulated mice mortality was recorded within 70 hous after intraperitoneal injection of LPS(20mg/kg)(n=37/WT,n=27/Hsp27Tg).The NF-kB activity for cardiac-tissue samples was analyzed by electrophoretic mobility shift assay(ENSA)and for cell culture samples by dual-reporter gene assay(n=4/group).The comparison of multiple groups was performed by one-way analysis of variance(ANOVA),and comparison of two groups was performed by Scheffe-test.Survival curves were analyzed by the log-rank test.P＜0.05 wns considered to be significant.Results The high expression of Hsp27 exhibited in myocardium of Hsp27 Tg,whereas not in myocardium of WT.LPS significantly reduced the cardiac function both in Hsp27 Tg and WT.However,compaled with LPS-treated WT,cardiac function was more significantly improved as evidenced by the increases of EF by 27.33%and FS by 37.09%(P＜0.01 or P＜0.05).Seventy hours after LPS injection,the mortality was 11.11% in Hsp27 Tg and 37.84% in WT.Compared with WT,the survival rate of Hsp27 Tg significantly increased(P＜0.05).The NF-kB activation was significantly inhibited by Hsp27(P＜0.01 or P＜0.05).Conclusions The high cardiac-specific expression of Hsp27 significantly inhibits cardiac dysfunction induced by endotoxemia,and at the same time improve the survival rate.The mechanism may be connected with Hsp27 downregulating NF-kB-activation induced by LPS.

Objective To observe the implementation of national food safety standard for “Iodine Concentration in Edible Salt”(GB 26878-2011) and its effectiveness on iodine nutritional status of key populations. Methods Information of iodine concentration in edible iodized salt of various provinces (autonomous regions and municipalities, including Xinjiang Production and Construction Corps) was collected using Baidu Searching Engine through the establishment of key words. Sal t samples were collected in Tianjin City and Aksu Region of Xinjiang , and the salt iodine concentration in both places was 30 mg/kg. In Tianjin, Hongqiao, Tanggu and Hangu, Beichen were selected as representatives of the downtown areas, the coastal areas and the suburbs, respectively and counties of Baodi and Ji were iodine deficiency areas in history. Sampling work was carried out from August 2012 to March 2013 in Tianjin. In Aksu, Yatuoer Township and Charqi Town in Baicheng County, Aotebeixi and Aketuohai Townships in Wushen County were chosen as iodine deficiency areas, and the survey was carried out from January to September 2013 . Random urine samples of school-age children ( 8 - 10 years old ) , pregnant women and lactating women were collected; urinary iodine was measured following the Method for Determination of Iodine in Urine by As3+-Ce4+ Catalytic Spectrophotometry(WS/T 107-2006) and iodine in edible iodized salt was measured following the General Test Method in Salt Industry Determination of Iodideion ( GB/T 13025 . 7-1999 ) . Results Fourteen of the provinces(autonomous regions and municipalities, including the Corps of Xinjiang) chose 25 mg/kg as their iodine concentration in edible iodized salt and 13 provinces chose 30 mg/kg. Besides, there were another 5 provinces providing 30 mg/kg particularly for pregnant women and lactating women while 25 mg/kg for other populations. In Tianjin, the medians of iodine concentration in edible iodized salt were ranged from 24.4 - 32.1 mg/kg in retail stores and 26.4 mg/kg at households. The household coverage rate of iodized salt and the proportion of households using adequately iodized salt were 78.5%(168/214) and 62.6%(134/214), respectively. The median ranges of urinary iodine were 178.2 - 183.9 μg/L in school children, 124.3 - 130.9 μg/L in pregnant women and 72.7 - 109.5 μg/L in lactating women. In Aksu, the medians of iodine concentration in edible iodized salt were 27.1 and 26.5 mg/kg in retail stores and households, respectively. The household coverage rate of iodized salt and the proportion of households using adequately iodized salt were 100.0% (363/363) and 98.9%(359/363), respectively. The median ranges of urinary iodine were 174.8 - 293.0, 154.9 - 230.0 and 135.8 - 239.3 μg/L among school children, pregnant women and lactating women, respectively. The median of iodine concentration in a special edible iodized salt sample reached 49.1 mg/kg, and qualified rate was 0(0/11) in Aksu. Conclusions All provinces , municipalities and autonomous regions ( including the Corps of Xingjiang ) in China have adjusted the iodine content in edible iodized salt in accordance with GB 26878-2011. However, in Tianjin the household iodine concentration in edible salt is lower than the local standards; the household coverage rate of iodized salt and the proportion of households using adequately iodized salt are lower than the national standards; pregnant women and lactating women are at risk of mild iodine deficiency.

Objective To compare treatment response according to the PERCIST1.0,RECIST1.1,EORTC,and WHO criteria in patients with colorectal liver metastases (CLM) who received neoadjuvant chemotherapy.Methods A total of 41 CLM patients (27 males,average age 68.48 years;14 females,average age 62.43 years) from January 2010 to September 2013 were included in this retrospective study.PET/CT scan was performed before chemotherapy and after 4-6 cycles′ chemotherapy.The baseline and the sequential follow-up 18F-FDG PET/CT of each patient were evaluated according to the PERCIST1.0,RECIST1.1,EORTC,and WHO criteria.The response was categorized into 4 levels including CR,PR,SD,PD.PET/CT images were used for both metabolic and anatomic evaluation.The concurrent diagnostic CT or MRI images (performed within 1 week of PET/CT) were also utilized when needed.The agreements of criteria were analyzed using Kappa test.The response rate (RR) and disease control rate (DCR) were compared using χ2 test.Results The RR and DCR according to the PERCIST1.0,EORTC and RECIST1.1 criteria were 31.71％(13/41) and 63.41％(26/41),31.71％(13/41) and 60.98％(25/41),17.07％(7/41) and 68.29％(28/41),respectively.The general comparison of PERCIST1.0 and RECIST1.1,EORTC and RECIST1.1 criteria showed good agreements (κ values: 0.711,0.689).Significant difference was not found in the DCR(χ2=2.000,P>0.05) but found in the RR(χ2=6.000,P<0.05) between PERCIST1.0 and RECIST1.1.Difference of DCR between EORTC and RECIST1.1 was not significant(χ2=3.000,P>0.05),while the RR had significant difference(χ2=6.000,P<0.05).The RR and DCR according to WHO criterion were 12.20％(5/41) and 70.73％(29/41),which had a good consistency with those according to PERCIST1.0 criteria (κ=0.629).Significant statistical difference was not found in the DCR(χ2=3.000,P>0.05) but found in the RR(χ2=8.000,P<0.05) between PERCIST1.0 and WHO criteria.Conclusions In evaluating CLM treatment response,anatomical criteria and metabolic criteria have a good consistency.But metabolic criteria are more sensitive for RR evaluating.

Objective:To predict and identify HLA-A * 0201 restricted CD8+ CTL epitopes in Mycobacterium tuberculosis (Mtb) antigen Ag85C, so as to provide evidence for epitope-based study for tuberculosis (TB) vaccine. Methods: The online database SYFPEITHI was applied to predict the potential HLA-A * 0201 restricted epitopes from Ag85C, an antigen of Mycobacterium tuberculosis. T2 cell line was used to assay the affinity between the predicted peptides and HLA-A * 0201 molecules. The specific CTL lines were induced from peripheral blood mononuclear cells (PBMCs) of HLA-A * 0201 positive TB patients and PPD+ healthy donors by peptides with high binding affinity to HLA-A * 0201 molecules. IFN-?production, in vitro proliferation and cytotoxicity of peptide-induced CTL were determined to screen HLA-A * 0201 restricted CD8+ CTL epitopes from those candidates. Results: Fourteen potential epitopes were identified from the SYFPEITHI database. After binding affinity assay, 3 of the 14 peptides (170-178 aa, 317-325 aa, and 144-153 aa) were found to have high binding affinity to HLA-A* 0201 molecules. However, only one peptide (144-153 aa) stimulated its specific CTL to release IFN-y, proliferate in vitro and produce specific cytotoxicity. Conclusion: We have successfully identified a HLA-A * 0201 restricted CD8+ CTL epitope of Mtb Ag85C-FLTREMPAWL( 144-153 aa) , which might be a candidate epitope for TB vaccine designing. Our findings provides a basis for developing novel and effective anti-TB vaccine.

Objective To study the activation induced cell death (AICD) of CD4+ T cells in primary biliary cirrhosis(PBC)murine model induced by poly I∶C. Methods Thirty female C57BL/6 mice were divided into model and control group randomly, and the former were injected with 5 mg/kg of poly I∶C, the later with PBS. PBC mice were detected 16 weeks after injection. CD4+ T cells isolated from spleen were stimulated in vitro by Con A and anti-CD3, and the apoptosis were determined by Annexin-V and PI staining. The expression of Fas, FasL and TRAIL were assayed by relative quantitative real-time PCR. Bcl-2 was detected by Western blot. Results Compared with control group, the portal areas of mice in model group were infiltrated with mononuclear cells obviously. The positive rate of serum antimitochondrial antibody(AMA) and the level of alkali phosphatase (ALP) were higher than that in control group (P<0.001). AICD of splenic CD4+ T cells in model group was lower than that of control group (P<0.001). The mRNA of FasL and TRAIL in model mice was down-regulated. Simultaneously, the anti-apoptosis protein Bcl-2 was up-regulated in model group. Conclusion These observations suggest that a defect in AICD of auto-reactive TH1 cells may contribute to the pathogenesis of PBC model. Furthermore, this defect in AICD may results from the change of Fas/FasL, TRAIL pathway and the up-regulation of Bcl-2.

Objective To investigate the cell phenotype for T cells in polyI: C induced primary biliary cirrhosis (PBC)animal model.Methods 20 female C57BI/6 mice,8 weeks old,were divided into model group and control group randomly. Mice in model group and control group were injected with polyI:C at a dose of 5 ms/ks and PBS,respectively.All mice were acrificed after 16 weeks after injection, and the sections of liver specimen were subjected to hematoxylin and eosin(H.E) staining.Serum AMA and ALP were detected.CD4+,CDs8+ and NKT cells in peripheral blood were determined by flow cytometry.The level of serum IL-4 and IFN-γ were assayed by EUSA.Results PBC mouse model was developed 16 weeks fter polyI: C injection. Infiltration of lymphocytes in portal area,positive serum AMA and high level of serum ALP were observed.The ratios of CD4+ T cells in model group and control group were(25.45±11.12)% and (26.72±0.63)%,respectively(t=0.314,P>0.05).The ratios of CDs+T cells in two groups were (18.3±0.91)% and (17.8±0.58)%,espectively(t=0.226,P>0.05).No significant change Was found for CD;and CDs+T cells in mice of both groups.However,NKT cells in peripheral blood of two groups were(11.56±5.09)% and (1.26±0.53)%,respectively(t=9.504,P<0.01).The number of NKT cells in model group was more than that of control group significantly.Simultaneously,serum L-4 and IFN·γ in mice of model group were also higher than that of control group.IL4 in senlm of two groups were (22.19±2.31)pg/ml and(8.72±0.87)pg/ml,respectively(t=58.06,P<0.01).IFN-γ in serum of two groups were(3.34±0.76)ng/ml and(1.14±0.21)ng/ml,respectively(t=23.31,P<0.01).Conclusions NKT cells increase greatly in eripheral blood of polyI:C induced PBC mouse model.NKT cells may play a critical role in the pathogenesis of PBC.

UNLABELLEDOBJECTIVE To study the effects of intra-abdominal pressure (IAP) on respiratory system, circulatory system and renal function in rats. To investigate the difference between the direct measure and indirect measure methods (via inferior vena cava and bladder) for IAP.

METHODSSixty Sprague-Drawly (SD) rats were randomly divided into 5 groups with different IAP (IAP value of 1-5 groups was 5, 10, 15, 20, 25 mm Hg,respectively) and healthy control group, 10 rats in each group. The parameters of respiratory system, circulatory system, renal function, and IAP value were recorded. The correlation between direct and indirect measurement methods was also analyzed.

RESULTSThere were no significant differences in above parameters between IAPI and healthy control groups. Compared with those in healthy control group, PaO2 significantly decreased (P < 0.05), SCr and BUN increased significantly in IAP2 group (P < 0.05). Other indexes in IAP3, IAP4 groups increased (P < 0.05 or P < 0.01) except for respiratory frequency(RF) and MAP (P > 0.05). PaO2 and MAP decreased (P < 0.01), and other indexes increased (P < 0.05 or P < 0.01) in IAP5 group. The values obtained from the indirect measure method were positively correlated with that from the direct measure method (r = 0.937, 0.955, P < 0.01, respectively).

CONCLUSIONSIAP can affect respiratory system, circulatory system and renal function in different degrees in rats. The indirect measure method can replace the direct measure method for IAP measure with little injuries.

Abdomen ; physiology ; physiopathology ; Animals ; Blood Pressure ; Cardiovascular System ; Compartment Syndromes ; Female ; Kidney Function Tests ; Male ; Pressure ; Rats ; Rats, Sprague-Dawley ; Respiratory System

To investigate the feasibility of using Real-Time PCR to evaluate the effectiveness of Sindbis virus inactivation by Methylene Blue with visible light. Sindbis virus was treated by Methylene Blue with different intensity of visible light and the transcribed cDNA was quantified by Real-Time PCR. Residual infectivity of treated virus was tested by cell infection method as parallel control at the same time. The residual infectivity of virus decreased from 6.50 lgTCID50/mL to under the limit of detection as light intensity increased. Meanwhile, the quantity of virus cDNA decreased significantly (P < 0.05), which correlated to the decline of virus infectivity (R2 > 0.98). Methylene Blue with visible light could cause lesion to nucleic acid of Sindbis virus, the extent of which was light intensity-dependent and correlated to the decrease of virus infectivity. The results demonstrated that Real-Time PCR can be a useful tool for evaluating effect of virus inactivation after Methylene Blue treatment with light.
Light ; Methylene Blue ; pharmacology ; Polymerase Chain Reaction ; methods ; Sindbis Virus ; drug effects ; genetics ; physiology ; radiation effects ; Virus Inactivation ; drug effects ; radiation effects