Swine influenza A virus belongs to the family of Orthomyxoviridae,which can cause upper respiratory tract infection in swine,avian and human.The influenza A virus that could not be sub-typed was detected from the respiratory tract specimens of patients with influenza-like symptoms in March 2009 in USA and Mexico,followed by identification of a novel swine-origin influenza A (H1N1) virus (S-OIV),which led to an epidemic outbreak in the above two countries and then spread all over the world,causing quite a few deaths.S-OIV differs from other influenza viruses in genotyping,antigenic characteristics,propagating characteristics,pathogenic characteristics and therapies.

Objective　To study efficacy of 2,4-dibromo-5, 5-d imethylhydantoin in killing vegetative forms of bacteria and spores of B. subtil is var.niger, and efficacy of 2,4-dibromo-5,5-dimethylhydantoin in dest roying antigenicity of HBsAg. Methods　Neutralizer test and efficacy of so lution of 2,4-dibromo-5,5-dimethylhydantoin in killing vegetative forms of ba cteria and spores. Neutralizer test and efficacy of solution of 2,4-dibromo-5, 5-dimethylhydantoin in destroying HBsAg antigenicity in suspension. Resul ts　The killing rate of Staphylococcus aureus and E. coli. was 100 % a fter exposure to its solution containing 4 mg*L-1 and 2 mg*L-1 av ailable bromo after 10 min and 20 min. The killing rate of spores of Bacil lus subtilis var. Niger also was 100% after exposure to its solution co ntainin g 2 000 mg*L-1 available bromo after 30 min. Its solution containing 1 0 00 mg*L-1 available bromo with could destroy HBsAg in su spension for 5 min. Conclusions　2,4-dibromo-5,5-dimethylhydantoi n can effe ctively kill vegetative forms of bacteria and spores of B. subtilis var.ni ger, and can completely destroy the antigenicity of HBsAg in the water.

The majority of current crisis intervention models are stage models.However,it is not certain that crisis intervention follows a linear process,and fixed procedure of stage models therefore may not provide efficient or effective crisis intervention.Task models,on the other hand,emphasize flexibility in crisis intervention by presenting main components,including three continuous tasks (i.e.,assessmem,safety,and support) and 4 focused tasks (i.e.,contact,re-establishing control,problem solving,and follow up).This theoretical framework may be helpful in creating standard practices of crisis intervention,examining the effectiveness of intervention plans,and providing guidance and training to counselors and researchers.

Objective To study the impacts of B7-H3 molecule on the proliferation of T lymphocytes in different activation conditions and on the secretion of relevant cytokines.Methods Peripheral blood samples were collected from healthy subjects to separate peripheral blood mononuclear cells (PBMC) by Ficoll density gradient centrifugation.T lymphocytes were isolated from some of the PBMCs and purified with T Cell Enrichment Kit.PBMC and purified T lymphocytes were activated by anti-CD3/CD28 monoclonal antibodies (McAb) in vitro.Flow cytometry analysis was used to detect the expression of CD28 and CTLA-4 on T lymphocytes at different time points for further analyzing the activation states of T lymphocytes.On this basis, human B7-H3-Fc fusion protein was added into the mixed co-cultivation system on days 0, 1, 2, 3 and 4, and Hu-Fc fusion protein was used as isotype control.CCK-8 method was performed to detect the proliferation of T lymphocytes in each group.ELISA method was used to detect the secretion of cytokines (IL-2, IL-10 and IFN-γ) and to analyze the immune responses induced by stimulating T lymphocytes at different states of activation with B7-H3.Results B7-H3 molecule significantly inhibited the quiescent T lymphocytes from secreting IL-2 and IL-10, but had no significant impact on IFN-γ secretion.Moreover, it significantly promoted the activated T lymphocytes to secret IL-2 and IFN-γ, but had no obvious impact on IL-10 secretion.Results of the cell proliferation assay showed that B7-H3 molecule inhibited the in vitro proliferation of T lymphocytes in the PBMC, but had no obvious impact on purified T lymphocytes.ConclusionThe regulatory effects of B7-H3 molecule on the immune functions of T lymphocytes vary with the activation states of T lymphocytes.

Objective To study the correlation between fractional excretion of uric acid (FEUA) and blood uric acid,body mass index (BMI),blood pressure,blood glucose,blood lipid and other metabolic factors in patients with primary gout.Methods Sixty-two patients with primary gout (gout group) and 32 healthy people (control group) were selected in this study.Gout group was divided into uric acid excretion decreasing group (FEUA ＜ 7％,29 cases),mixed group (7％ ≤FEUA ≤ 12％,25 cases) and uric acid production increasing group (FEUA ＞ 12％,8 cases) according to the level of FEUA.The fasting blood glucose (FPG),2-hour postprandial blood glucose (2 h PBG),blood lipid,serum creatinine,blood uric acid,glycosylated hemoglobin were tested.24 hours urine was collected and urinary uric acid and urinary creatinine was measured,FEUA was calculated and analyzed.Results BMI,mean arterial pressure,blood uric acid,glycosylated hemoglobin,total cholesterol,2 h PBG in gout group was higher than that in control group,and high density lipoprotein cholesterol,FEUA was lower than that in control group,and there was significant difference (P ＜ 0.05).There was no significant difference in age,FPG,low density lipoprotein cholesterol,triacylglycerol between two groups (P＞ 0.05).There was no significant difference in age,blood uric acid,FPG,2 h PBG,glycosylated hemoglobin,total cholesterol,low density lipoprotein cholesterol,high density hpoprotein cholesterol among uric acid excretion decreasing group,mixed group and uric acid production increasing group (P ＞ 0.05),and there was significant difference in BMI,mean arterial pressure,triacylglycerol,FEUA among three groups(P＜ 0.05).FEUA was negatively correlated with blood uric acid in control group and gout group (r =-3.900,-0.476,P ＜0.05).FEUA was positively correlated with 24 h urinary uric acid in gout group (r =0.465,P =0.001),and nagatively correlated with triacylglycerol (r =-0.304,P ＜ 0.05).Pearson analysis showed that FEUA was negatively correlated with blood uric acid in uric acid excretion decreasing group (FEUA ＜ 7％) (r =-0.392,P ＜ 0.05),FEUA was positively correlated with blood uric acid in non uric acid excretion decreasing group (FEUA ≥7％)(r =0.437,P ＜ 0.05),but 24 h urinary uric acid was not correlated with blood uric acid(P ＞ 0.05).Multi-stepwise regression analysis showed that blood uric acid,glycosylated hemoglobin,FEUA was significantly correlated with the onset of the gout (P ＜ 0.05).Conclusions Besides blood uric acid level,there are significant changes in primary gout in blood pressure,serum glucose and lipid levels.FEUA could be used to estimate the ability of renal excrete the uric acid.Mean arterial pressure,glycosylated hemoglobin and FEUA are the risk factors for gout.

Objective To investigate the value of B7-H3 in expressed prostatic secretions (EPS) in differential diagnosis of patients with inflammatory elevation of PSA in t-PSA gray zone (4-10 ng/ml). Methods One hundred and sixteen patients from the ages of 19 to 80 years (mean, 40 years) were stu-died. In the group there were 91 chronic prostatitis (CP) patients (mean age 31 years, 19-49 years), including 11 chronic bacterial prostatitis (type II) patients, 26 inflammatory nonbacterial prostatitis (IIIA) patients and 54 noninflammatory nonbacterial prostatitis (IIIB) patients. Transrectal ultrsound guided prostate biopsy was performed on 25 patients (mean age 71 years, 62-80 years) with t-PSA in gray zone (7.21±2.60 ng/ml). Five had positive results, Gleason score was 6 in two cases, 7 in two cases and 8 in one case. Twenty patients had negative results, of whom 11 patients had inflammatory cell infiltration. EPS was collected by transrectal massage, and Enzyme-linked immunosorbent assays (ELISA) were performed for B7-H3 detection. In addition, 11 normal male controls with a mean age of 30 years (24-46 years) were recruited into the study. Volunteers were excluded if they had a history of genitourinary symptoms or surgery.Results The EPS B7-H3 levels of controls, II, IIIA, IIIB groups were 49.81±11.54, 19.33±13.90, 17.67±15.76, 25.14±13.44 ng/ml, respectively. The levels of EPS B7-H3 in positive biopsy, noninflammatory negative biopsy and inflammatory negative biopsy groups were 26.30±16.32, 30.23±18.42, 10.11±5.42 ng/ml, respectively. The highest levels were found in the control group (P<0.01). Compared to the IIIB, B7-H3 levels in II and IIIA groups were significantly lower (P<0.05). There was no significantly difference between II and IIIA groups (P>0.05). The EPS B7-H3 levels in the inflammatory negative biopsy group were statistically lower than in positive biopsy and noninflammatory biopsy groups (P<0.05). But no significant differences were found among inflammatory negative biopsy, II and IIIA groups (P>0.05). Receiver operating curve (AUC=0.883, P=0.001) utilizing EPS B7-H3 levels≤16.24 ng/ml identified patients with inflammatory elevation of PSA with a sensitivity of 78.6% and a specificity of 81.8% from patients with t-PSA in gray zone. Conclusion The EPS B7-H3 detection provides a new way for differential diagnosis of patients with inflammatory elevation of PSA in t-PSA gray zone resulting in a reduction of unnecessary prostate biopsy.

Objective To study characteristics and immune mechanisms of CD11b+ GR-1-myeloid-derived suppressor cells (CD11b+ GR 1+ MDSCs) in elderly mice,as compared with those of healthy young mice.Methods Totally 20 healthy C57BL/6 young mice (aged 1-2 months) and 20 elderly mice (aged over 18 months) were randomly chosen and splenetic CD11b+ GR-1+ MDSCs were sorted with the MDSCs Isolation Kit.In vitro assays,the effects of young and elderly CD1 1b+ GR 1+ MDSCs on the proliferation of T cells were determined by Brdu Elisa.Transwell co-culture and real-timePCR were used to identify the mechanisms of different immune suppressive functions of CD11b+GR 1+ MDSCs sorted from young mice and elderly mice.Results Compared with young MDSCs,elderly MDSCs could evidently suppress the proliferation of T cells (t=8.67,P＜0.001),and this function could be reversed by trans-well co-culture (t=6.93,P＜0.001).The results of realtime PCR revealed that,compared with young MDSCs,elderly MDSCs expressed higher levels of arginase-1 (ARG-1),inducible nitric oxide synthase (iNOS),reactive oxygen species (ROS),interleukin 10 (IL-10),IL13 and transforming growth factor (TGF)-β (t=9.04,4.86,7.04,6.92,4.51,5.46,respectively,P＜0.05 or P＜0.01).Conclusions CD11b+GR-1+MDSCs sorted from healthy elderly mice can evidently suppress the proliferation of T cells through cell-cell contact and secretion of suppressive medium.

Objective To investigate the level of subpopulations of myeloid-derived suppressor cells (MDSCs) in elderly tumor-bearing mice versus elderly tumor-free mice,and to study the difference in immune suppressive functions between different subpopulations and their mechanisms.Methods A total of 20 healthy C57BL/6 elderly mice(aged 18-20 months) were randomly chosen to establish Lewis lung cancer models.The amount of monocytic-MDSCs (MO-MDSCs) and polymorphonuclear granulocytic-MDSCs(PMN-MDSCs) in tumor-free and tumor-bearing elderly mice was evaluated by using flow cytometry.MO-MDSCs and PMN-MDSCs were separated with Magnetic-Activated Cell Sorting (MACS) MicroBeads and their morphological characteristics were observed after May-Grunwald-Giemsa staining.The effects of MO-MDSCs and PMN-MDSCs on the proliferation of T cells were determined by Brdu-enzyme-linked immunosorbent assay(ELISA).And the immune suppressive mediators secreted by the subpopulations were detected by Real-time polymerase chain reaction(PCR).Results Compared to the tumor-free group,the proportion of MO-MDSCs in the spleen of tumorbearing group were increased [(12.44± 1.20) ％ vs.(38.42±3.66) ％,t=5.67,P＜0.001],while PMN-MDSCs were not [(10.34±0.68) ％ vs.(12.18±1.27) ％,t=2.21,P=0.09].The result of Brdu-ELISA showed that MO-MDSCs could suppress the proliferation of T cells [(0.30 ± 0.18) vs.(3.38±0.96),t=8.33,P＜0.001],while PMN MDSCs could not [(2.69±0.45)vs.(3.38±0.96),t =1.72,P=0.11].The result of PCR showed that as compared with PMN-MDSCs,Mo-MDSCs had the increased expression levels of arginase-1 (ARG-1),inducible nitric oxide synthase (iNOS),interleukin-10 (IL-10),interferon-γ(IFN-γ) (t =4.31,8.89,1.70,3.13,respectively,P ＜ 0.01 or 0.05),while the expression levels of interleukin-13 (IL-13),transforming growth factor-β(TGF-β) had no differences (t=4.94 and 2.75,P =0.39 and0.47).Conclusions MO-MDSCs are significantly increased in elderly Lewis lung cancer mice models.MO-MDSCs could mediate lung tumor evasion by suppressing the proliferation of T cells through highly expressing ARG-1,iNOS,IL-10 and IFN-γ.

Objective To investigate the feasibility of using leukocytes that were filtered out by LeukoReduction System ( LRS) to replace conventional human peripheral blood leukocytes in experimental researches and to comparatively analyze the differences between them in vitro biological functions and pheno-types of T cells. Methods Mononuclear cells were isolated from LRS-separated leukocytes and whole blood sample that collected from the same person by using Ficoll. Fluorescence-activated cell sorting ( FACS) was performed to analyze the phenotypes of T cells. CD3+T cells were sorted out by using magnetic beads. The T cells that were collected by using two different ways were incubated with anti-CD3 and anti-CD28 antibodies and IL-2 in vitro for 10 days. Several assays including cell counting, FACS and cytometric beads array ( CBA) were performed to comparatively analyze the differences in biological functions and phenotypes of T cells that were isolated by different methods. Results The phenotypes of T cells isolated from LRS filter and whole blood sample were highly similar at the initial stage. The sorting rate of CD3+T cells form LRS filter reached a high level and met the requirements for experimental researches. No statistically significant differ-ences in cell count, phenotype, expression of costimulatory molecules and cytokine secretion were observed between T cells isolated from LRS filter and whole blood sample. Conclusion This study suggested that the T cells isolated from LRS filter could be used as an alternative to whole blood T cells for fundamental resear-ches since they were similar in cell vitality, phenotype and biological functions. It provided a new way to solve the problem of blood shortage in clinic and scientific research.

Objective To study on the expression and clinical significance of programmed death ligant 1 (PD-1) in the development of colorectal cancer.Methods Flow cytometry (FCM) was used to detect and analyse the positive expression rate of PD-1 on CD3+ T cells in peripheral blood samples of 88 colorectal cancer patients and 74 healthy volunteers,and in sixpairs of tissue specimens (colorectal cancer tumor and adjacent normal tissues).Colon cancer tumor-bearing mice models were constructed by subcutaneous implantation of murine colon cancer cell line CT26,and the changes of positive expression rate of PD-1 on CD3+ T cells in spleens and transplanted tumor tissues were also detected by FCM in different time points during five,eight,11,14,17,20 days after the model was constructed.Finally,the experimental results were analyzed by t test or variance analysis.Results The positive expression rate of PD-1 on CD3+ T cells in peripheral blood of colorectal cancer patients was significantly higher than that in healthy volunteers ((29.25 ± 9.37) ％ vs (19.35 ± 7.37) ％,t =7.375,P＜ 0.01),and the positive expression rates were significantly increased in colorectal cancer patients with lower degree of differentiation,lymph node metastasis and higher Duke's stage.The positive expression rate of PD-1 on CD3+T cells was significantly higher in colorectal cancer tissues compared to adjacent normal tissues ((52.38±12.28)％ vs (24.72±4.78)％,t=5.143,P＜0.01).In the colon cancer tumor-bearing mice models,along with the growth of transplanted tumors,PD-1 positive expression rates on the CD3+T cells of tumor-bearing mice spleens and transplanted tumors showed a gradually rising tendency.From the eighth day to the 20th day after subcutaneous transplanted colon cancer cells,the average positive expression rate of PD-1 on the CD3+T cells of tumor-bearing mice spleens rose from (52.83±6.17)％ to (77.30± 6.84) ％,and the average positive expression rate of transplanted tumors rose from (60.43 ± 2.77)％ to (90.47±4.31) ％.Conclusion There is a correlation between the expression of PD-1 on mature T cell surface and the development of colorectal cancer,and the high specific expression of PD-1 may promote the invasion and metastasis of colorectal cancer.