Objective:To evaluate the role of IL-23 in the production of IFN-?, cell subsets and regulation by human peripheral blood mononuclear cells(PBMCs).Methods:PBMCs were isolated from normal human peripheral blood and cultured with IL-23 in different culture conditions. The level of IFN-? in the culture supernatants was examined by ELISA. The subsets and frequency of IFN-?-producing cells were examined at a single cell level by flow cytometry.Results:IL-23 could directly induce IFN-? production by PBMCs and have synergistic effect with IL-2 on the induction of IFN-? production in a dose dependent manner. The data from Flow cytometric analysis indicated that IL-23 could induce IFN-? expression by CD3~-CD56~+NK cells but not CD3~+CD4~+ and CD8~+T cells. It is noted that IL-23 predominantly induced IFN-? expression by NK cells with high expression of CD56 molecules. Addition of Th2 cytokines or anti-IL-12R?1 mAbs resulted in the inhibition of IL-23-inducing IFN-? production.Conclusion:IL-23 can directly induce IFN-? production by PBMCs. The induction of IFN-? induced by IL-23 can be suppressed by Th2 cytokines and anti-IL-12R?1 mAbs. The data indicated that Th2 cytokines and anti-IL-12R?1 mAbs might have the potential application for the treatment of IL-23-mediated autoimmune diseases.

AIM: To study the effect and mechanism of BCG on human nature killer cells. METHODS: PBMC or purified NK cells were isolated from normal human peripheral blood with negative anti-Mycobacterium tuberculosis antibody and cultured with BCG, IL-12, BCG plus IL-12 and BCG plus anti-IL-12Rβ1 mAb (2B10), respectively. The levels of IFN-γ and IL-12p40 in the culture supernatants were measured by ELISA. The frequency of IFN-γ and granzyme B producing cells were analyzed by ELISpot. The cytolytic activity was detected by MTT reduction assay. The surface expression of IL-12Rβ1 on NK cells was detected by flow cytometry. RESULTS: BCG significantly induced IFN-γ production by PBMC in a dose-dependent manner. When PBMC was stimulated with BCG, the frequency of granzyme B producing cells was higher than that in unstimulated PBMC (P<0.05). BCG enhanced the cytotoxic activity of PBMC. BCG alone didn' t induce IFN-γ production by purified NK cells, but it can augment IL-12-induced IFN-γ production by purified NK cells. The cytotoxic activities of BCG-stimulated and unstimulated purified NK cells were not significantly different (P>0.05). BCG induced IL-12 production by PBMC in a dose-dependent manner and enhanced IL-12Rβ1 expression on different subsets of NK cells. Blocking the effect of IL-12 by anti-IL-12Rβ1 mAb (2B10) inhibited BCG-induced IFN-γ production and granzyme B releasing by PBMC. CONCLUSION: BCG can indirectly promote biologic activity of NK cells and the production of endogenous IL-12 combined with up-regulation IL-12Rβ1 expression on the surface of NK cells is a part of the mechanisms of IL-12 on human NK cells.

Objective To investigate the effects of metformin on plasma ghrelin and peptide YY (PYY)levels in newly-diagnosed type 2 diabetic patients,and to study the impact of metformin on body weight.Methods A prospective nested case-control study was designed as a research protocol.Sixty four newly-diagnosed type 2 diabetic patients were treated with metformin for 12 weeks.The patients were divided into two groups:weight loss group and non-weight loss group according to the changes in body weight after metformin treatment.Fasting plasma ghrelin and PYY levels and other metabolic parameters were measured before and after metformin treatment.ResultsFasting plasma ghrelin level was significantly decreased in the patients after metformin treatment [ ( 10.71 ±2.68 vs 11.81 ±3.19 )ng/ml,P＜0.05 ].Fasting plasma PYY level was significantly increased in patients after metformin treatment [ ( 136.86+39.14 vs 128.42+37.31 ) pg/ml,P＜0.05 ].After metformin treatment,43.7％ of the patients lost body weight significantly.Fasting plasma ghrelin level was decreased by 16.6％ after treatment in the weight loss group,as compared with 6.2％ in non weight loss group( P＜0.05 ).Fasting plasma PYY level was increased by 10.8％ after treatment in the weight loss group,as compared with 3.5％ in the non-weight loss group (P ＜ 0.05 ).Conclusions The fasting plasma ghrelin level in the weight loss group was lowered more significantly compared with that in the non-weight loss group after metformin treatment.The fasting plasma PYY level in the weight loss group was elevated more significantly as compared with that in the non-weight loss group after metformin treatment.The mechanism remains to be further studied.

Objective To investigate effect of the transtheoretical model and stages of change (TTM) on self-management paradigm,serum calcium and phosphorus levels in patients undergoing maintenance hemodialysis (MHD).Methods Total 140 MHD patients were separated into two groups as the intervention group (65 of 70 cases completed with TTM) and the control group (63 of 70 cases completed with routine nursing care) by random number table.Data obtained by using hyperphosphatemia related knowledge questionnaires and Self-management Rating Scale before and after intervention.The levels of serum calcium and phosphorus between the above groups were compared.Results The scores of self-management behavior were higher in control group after 3 months intervention,the differences was statistically significant (P ＜ 0.05),in which the scores of problem solving (2.97 ± 0.28),self care activities (2.99±0.28),relationships (3.13±0.35) and emotional treatment (2.80 ± 0.32) in experimental group was apparently higher than control group at the same time,the scores were (2.63±0.32),(2.67±0.38),(2.76±0.42),(2.44±0.36) respectively in conrtol group,t values were-0.907,-7.68,-6.03 and-8.43,respectively.And also,there was statistically significant difference between the two groups in serum phosphorus level [(1.81±0.49) mmol/L vs.(2.56±0.69) mmol/L],the product of calcium and phosphorus [(55.33±14.45) mg2/dl2 vs.(72.35±20.94) mg2/dl2],serum parathyroid hormone [238.00 (130.35,413.85) ng/L vs.297.75 (155.02,760.37) ng/L] (t=11.01,8.43,Z=-2.09,P＜0.05).Conclusions TTM is conducive to promote behavior change,improve self-management conduct,and also reduce serum phosphorus level in MHD patients.

Objective To investigate the distribution and sequence conservation of Hap adhensin encoding gene (hap) in clinical isolates of nontypeable Haemophilus influenzae (NTHi), to screen out and identify the predominant T-and B-cell (T-B) combined antigenic epitopes on Hap protein and to analyze their immunogenicity.Methods Sequence conservation of hap genes in NTHi strains and T-B combined antigenic epitopes were predicted using bioinformatic softwares.PCR was used to amplify the 156 bp segment at 5′-end and the 855 bp segment at 3′-end of hap gene (hap-5′-156 and hap-3′-855) and the amplified products were sequenced.Phage display systems of seven T-B combined antigenic epitopes located on the 55 aa segment at N-terminal and the 285 aa segment at C-terminal of Hap protein (Hap-N52 and Hap-C285) were constructed.Western blot assay and ELISA were performed to detect the antigenicity and immunoreactivity of different T-B combined epitopes displayed by recombinant phage PⅢ protein (rPⅢ).Results Hap protein encoded by the hap gene in NTHi was located on membrane surface.Sequences of the 156 bp segment at 5′-end and the 855 bp segment at 3′-end of hap genes extracted from different NTHi strains were relatively conservative, but many mutations were found in sequences at the middle regions of these hap genes.All of the 56 NTHi strains carried hap-5′-156 and hap-3′-855 segments and shared 92.3％-100％ identities in nucleotide and amino acid sequences of these segements.Hap-N5-24 in the Hap-N52 segment as well as Hap-C4-27, Hap-C28-47, Hap-C114-129, Hap-C150-173, Hap-C200-227 and Hap-C241-267 in the Hap-C285 segment was predicted as the T-B combined antigenic epitope with a higher score and less mutations.Results of Western blot assay and ELISA confirmed that the rPⅢ-displayed Hap-C4-27 and Hap-C150-173 epitopes presented clear hybridization bands with NTHi antisera, and 96.9％ (63/65) and 92.3％ (60/65) of serum samples from children with NTHi infection were positive for antibodies against Hap-C4-27 and Hap-C150-173 epitopes, respectively.Conclusion The gene of hap is widely distributed in clinical isolates of NTHi.Moreover, sequences of the 156 pb segment at 5′-end and the 855 bp segment at 3′-end of hap gene are conservative.Hap-C4-27 and Hap-C150-173 are the predominant T-B combined antigenic epitopes on Hap protein, suggesting that they can be used as epitope candidates for developing multiple antigenic peptide vaccines against NTHi.

Objective To study protein variation between PtenL/LMEFs and Pten△/△MEFs cells.Methods Two-dimensional electrophoresis(2-DE) was employed to compare the differential expression proteins between PtenL/LMEFs and Pten△/△MEFs cells.Six differential expression proteins were digested in gel by enzyme and the mass of generated peptides was measured by matrix assisted laser desorption ionization time of flight mass spectrometry(MALDI-TOF-MS).The data obtained from peptide mass fingerprinting(PMF) were searched using the Internet available database and five proteins were identified.Results Compared with that of PtenL/LMEFs,expression level of proteins including phosphoglycerate mutase 1(PGAM1) and peptidyl-prolyl cis-trans isomerase C(PPIC) was up-regulated,whereas expression level of Transgelin 2 was down-regulated in Pten△/△MEFs cells.B and F proteins were both identified to be peroxiredoxin-6.They had similar molecular weight but different PI which might be caused by post-translation modification.B protein was only expressed in Pten△/△MEFs cells.Conclusion The protein profile of Pten△/△MEFs cells displayed obvious difference compared to that of PtenL/LMEFs cells.The results implied that various distinct different proteins might lead to cancer.

Unfavorable psychic reactions or psychological crisis always occur in patients preliminarily diagnosed with cancer.By analyzing the psychological activities and affecting factors,we try to discuss the measures of psychological crisis intervention for patients preliminarily diagnosed with cancer.

Objective:To elucidate the characterization of antigen-specific memory T cells from PPD~+ individuals after stimulation with BCG in vitro.Methods:PBMCs were isolated from PPD~ -/+ normal human peripheral blood and stimulated with BCG. The level of IFN-? in the culture supernatants was assayed by ELISA and the frequency of IFN-?-producinging cells was detected by ELISPOT. The subsets and frequency of cytokine-producing cells were determined at a single cell level by flow cytometry.Results:After stimulation with BCG, PBMCs from PPD~+ but not PPD~- individuals produced significantly high levels of IFN-? in culture supernatants detected by ELISA(P

Objective To explore the relation of the expressions of MDM2 and VEGF in osteosarcoma with the pathological parameters and prognosis of the tumor.Methods The expressions of MDM2 and VEGF were detected with immunohistochemical(SP) method in specimens from 56 cases of osteosarcoma.The correlation between the expressions of MDM2,VEGF and pathological grade,metastasis and prognosis was analyzed statistically.while 8 cases of fibrous dysplasia of bone were used as negative control group.Results The positive rates of MDM2 and VEGF in osteosarcoma were 64.3%(36/56) and 67.9%(38/56),respectively .MDM2 and VEGF didn't express in negative control group.The expression of MDM2 and VEGF were not significantly correlated to the pathological grades of the osteosarcoma,but which were significantly correlated with tumor metastasis and prognosis(P

This paper introduced the framework and approach, explained the new categories, and elaborated the implementation of International Classification of Functioning, Disability and Health (Children and Youth Version) (ICF-CY) in fields of rehabilitation for children with disabilities. The publishing and distribution of ICF-CY international Chinese version would be a big thing for the Chinese to implement ICF in related fields.