Objective Assaying HBV cccDNA in periphral blood lymphocyte and discuss its clinical significance.Methods Using lymphocyte isolating solution to isolate lymphocytes in the peripheral blood, and quantitative florescent realtime PCR was used to detect HBV cccDNA.Results 29 cases of 36 CAH patients was detected positive(80.56%),32 cases of 61 CPH patients was detected positive(52.46%) and 1 case of 59 ASC was detected positive(0.02%) respectively.All 47 healthy people specimen was found negative by the method.CAH group,CPH group and ASC & negative control group show significance statistically(P

Objective To investigate whether nuclear transcription factor κB(NF-κB) through Toll-like receptors 2(TLR2) was activated by lipid-associated membrane proteins(LAMPs) of Mycoplasma genitalium.Methods LAMPs were extractded and THP-1 cells were stimulated.The activation of NF-κBp65 was detected by ELISA and the expression of TLR2 mRNA was detected by RT-PCR.Effects of TLR2 neutralizing antibody on LAMPs induced the activation of NF-κBp65 was analyzed by ELISA.After LAMPs stimulated 293T cells with the co-transfection pFLAG-TLR2,pNF-κB-luc,pRL-TK,the activity of NF-κB firefly luciferase and pRL-TK Renilla luciferase were detected by the dual-luciferase reporter gene,to analyzed the role of TLR2-mediated NF-κB activation by LAMPs in 293T cells.Results The activation of NF-κBp65 was mediated in LAMPs induced THP-1 cells and was significantly increased by LAMPs in a dose dependent manner.when LAMPs was 4.0 μg/ml,the activation of NF-κBp65 was the highest level.TLR2 mRNA expression was up-regulated by LAMPs in THP-1 cells.TLR2 neutralizing antibody could inhibit the activation of NF-κB by 60% in LAMPs stimulated THP-1.NF-κB fluorescence was significantly increased by co-transfection pFLAG-TLR2 in a dose-dependent manner. ConclusionMycoplasma genitalium-derived lipid-associated membrane proteins activate NF-κB via TLR2 and the activation of TLR2-mediated play an important role in pathogenic process of LAMPs.

In this paper, a remote monitoring system based on three tiers framework of ″Home-Community-Hospital″ for home monitoring is introduced. Its characteristics include multiuser concurrency operation, multiple testing parameter, real-time interactivity, dependable and secure data communication, high-speed link with database and load balancing. According to user's requirements, various applications can be developed to perfect medical information system based on B/S mode. A portable multiparametric tester based on embedded system has the functions of wireless data transmission and network communication, whose hardware and software designs are modularized.

This thesis analyzed the inevitability of medical negligence according to the nature of medical behavior,particularity of medical profession and psychological factors of medical staff.How to control the occurrence of medical errors is an important social problem which is the whole community has been looking forward to solve.This thesis proposed methods to reduce medical negligence from the aspects of service concept,medical education and healthy psychology.

Quality is the lifeline of graduate education and the tutor team plays an important role in postgraduate training. To control the selection of tutors strictly, to implement the combination of openness and stability in tutor team, and to carry out academic exchanges actively are the keys to construct an excellent tutors staff and to ensure the quality of graduate student training.

To investigate the effects of CD14 on nuclear transcription factorκB (NF-κB) was activated by lipid-associated membrane proteins (LAMPs) of Mycoplasma genitalium (Mg) ,THP-1 cells were pretreated with serum human or CD14 neu-tralizing antibody ,and then were stimulated by LAMPs .The activation of NF-κBp65 was detected by ELISA .After LAMPs was pretreated with sCD14 stimulated Hela cells with the co-transfection ,the activity of NF-κB luciferase were detected by the dual-luciferase reporter gene to analyze the role of CD14-mediated NF-κB activation by LAMPs .The activation of NF-κBp65 was significantly up-regulated in LAMPs activated THP-1 cells by human serum .It’s suggested that CD14 neutralizing anti-body could inhibit the activation of NF-κBp65 in LAMPs stimulated THP-1 .The activation of NF-κB was significantly up-regu-lated in LAMPs activated Hela cells by mCD14 or sCD14 .CD14 could augment the activation of NF-κB by Mg LAMPs .

ObjectiveTo screen a 12-mer phage display peptide library by the polyclonal antibody (pAb) against the recombinant adhesion protein of Mycoplasma genitalium (rMgPa) in order to obtain the antigenic mimic epitopes of MgPa.MethodsThe purified pAb was used to screen the immunodominant mimic epitopes of MgPa by a random 12-peptide phage display library.Seventy-four recombinant phage clones were randomly selected,and then DNA sequence analysis and computer-based bioinformatics analysis were performed to define the consensus amino acid residues of the mimotopes by MIMOX.The binding specificities of the selected phage-displayed peptides to the purified pAb were confirmed by ELISA,competitive ELISA and Western blot analysis.Results After four rounds of biopanning,a significant enrichment of phages was achieved,the inserts from 74 phage clones distinguished 45 peptides based on the different amino acids sequences.Amongst 45 peptides,36 peptides were ELISA positive and 23 peptides that absorbance values were higher than 1.5 showed high reactivities with pAb and effectively inhibited the binding of pAb to rMgPa.Immunoscreening via phage display peptide library revealed three different mimptopes of adhesion protein of M.genitalium,P-S-A-A/V-X-R-F/W-E/S-L-S-P,A-K-I/L-T/Q-X-T-L-X-L and K-S-L-S-R-X-D-X-I.Results of bioinformatics analysis by MIMOX demonstrated that S,A,F for cluster 1,A,K,I,T and L for cluster 2,K,S,L,R,D and I for cluster 3,may be the key consensus amino acid residues in the aligned mimotopes,respectively.ConclusionAntigenic mimics on MgPa were successfully identified and the motif P-S-A-A/V-X-R-F/W-E/S-L-S-P,A-K-I/L-T/Q-X-T-L-X-L and K-S-L-S-R-X-D-X-I may represent the immunodominant mimic epitopes of MgPa.And S,A,F K,I,T,L,R and D may be the key amino acid residues for the epitopes of MgPa.

Objective To provide experimental evidence for the development of multi-epitope-baseded marker vaccines through investigating the humoral and cellular immune responses in BALB/c mice induced by the multiple antigen peptides (MAPs) with the mimic epitope.Methods Three types of MAPs in eight branched forms containing the mimic epitope of Mycoplasma genitalium adhesion protein (MgPa) were prepared using poly-lysine as the core matrix.The purity of MAPs was analyzed by reverse phase high performance liquid chromatography (RP-HPLC).The molecular weights of MAPs were characterized by Mass Spectrometry.The BALB/c mice were immunized intramuscularly for four times with single or mixed MAPs.The specific IgG antibody and the subtype of IgG antibody in serum of the immunized mice were detected by indirect ELISA.The proliferative responses of the spleen lymphocytes were detected using MTT assay.The ELISA were used to detect IFN-γ and IL-4 levels in the cultured supematant of spleen lymphocytes.Results The three types of MAPs containing the mimic epitopes were successfully prepared with high purity.They,could stimulate mice to produce specific IgG antibodies,of which,the major antibody isotype was Th1 immune response-associated IgG2a.Compared with the single MAP immunization group,the mixed-MAPs immunized mice produced more IgG,IgG1 and IgG2a antibody (P＜0.05).Furthermore,these MAPs could enhance the specific proliferation of spleen lymphocytes in immunized mice and induce the production of IFN-γ and IL-4.The levels of IFN-γand IL-4 in mixed-MAPs group were significantly higher than those of the single MAPs group (P＜0.01).Conclusion The three types of MAPs could induce strong specific cellular and humoral immune responses.The immunological competence of the mixed-MAPs was stronger than those of the single MAP.

Objective To investigate whether macrophage-activating lipopeptide-2 ( MALP-2) in-duces the expression of heme oxygenase-1 ( HO-1 ) in THP-1 cells and its possible mechanism .Methods Human monocyte cells THP-1 were cultured in vitro and then were incubated with various concentrations (0, 0.01, 0.1, 1.0 or 5.0 ng/ml) of MALP-2 for 16 h, or were stimulated by 5.0 ng/ml MALP-2 for different length of time (0 h, 4 h, 8 h, 12 h, 16 h or 24 h).The expression of HO-1 at mRNA and protein levels were detected by real-time PCR analysis and Western blot assay .The enzyme activity of HO-1 was detected by colorimetric analysis.THP-1 cells were pre-incubated with 30 μmol/L of SB203580, PD98059 and SP600125 for 30 min and then were cultured with 5.0 ng/ml MALP-2 for 16 h to investigate the role of mito-gen-activated protein kinases (MAPKs) signaling pathway in HO-1 production.After incubating THP-1 cells with 5.0 ng/ml MALP-2 for different periods of time, NF-E2-related factor 2 (Nrf2) protein was detected by Western blot assay to study the effects of Nrf2 pathway on MALP-2-induced HO-1 expression.Nrf2 and HO-1 proteins were measured by Western blot assay after transfecting THP-1 cells (1×106/well) with Nrf2 siRNA at a final concentration of 100 nmol/L.Results MALP-2 enhanced the expression of HO-1 at mRNA and protein levels as well as the enzyme activity of HO-1 in THP-1 cells in a concentration-dependent manner.The expression of HO-1 protein induced by MALP-2 was significantly inhibited by 30 μmol/L MAPKs specific inhibitors ( SB203580 , PD98059 and SP600125 ) .MALP-2 induced Nrf2 translocation at a concentration of 5.0 ng/ml.The expression of Nrf2 and HO-1 proteins were significantly decreased in Nrf 2 siRNA-transfected THP-1 cells.Conclusion MAPKs and Nrf2 signaling pathways were involved in the MALP-2 induced HO-1 expression .

Objective:To observe the molecular mechanism involved in expression of hemeoxygenase -1 (HO-1) induced by a macrophage-activating lipopeptide-2 (MALP-2).Methods:THP-1 cells were cultured in vitro and stimulated by MALP-2 for 12 h, expression of HO-1 was detected by Western blot .TLR2 and TLR6 neutralizing antibodies incubation , dominant negative plasmids transfection were used to assess the functional of TLR 2,6 in mediating HO-1 expression.Phosphorylation of c-Src and Akt were detec-ted by Western blot, and c-Src siRNA and PI3K inhibitor LY294002 were used to investigate the role of c-Src and PI3K in HO-1 ex-pression.Results:MALP-2 induced c-Src phosphorylation , and TLR2 and TLR6 neutralizing antibodies , or their dominant negatively plasmids could abrogate this effect .In addition, siRNA of c-Src could decrease the phosphorylation level of Akt , and the PI3K inhibi-tor could inhibit HO-1 expression.Conclusion: MALP-2 can induce THP-1 cells expression of HO-1 through TLR2,6/c-Src/PI3K pathways .