The pathogenesis of immune dysfunction in septic has been summarized in this review.Innate immune response toward pathogens is initiated by pattem recognition receptor (PRR) such as Toll-like receptor (TLR) .Inflammatory cytokines derived from innate immune response not only cause inflammatory response.but also trigger adaptive immune responses through inducing the differentiation of naive T cell into Th1, Th2, CD4~+CD25~+Foxp3~+ regulatory T cells (Treg), and Th17 cells. Adaptive immune response might suppress or enhance inflammatory response.Abnormal activation of innate/adaptive immune responses may coexist in sepsis, resulting in immune dysfunction.Logical and adequate approach of immunoregulation therapy to sepsis may be to suppress PRR persistent activation by eliminating endogenous or exogenous ligands, as well as to avoid excessive inhibition of immune responses to infection.

Objective　To explore the mechanism of enhancing killing activities of tumor cell-specific cytotoxic T lymphocyte (CTL) by IL-4 gene modification. Methods　IL-4 gene was introduced into human hepatocellular carcinoma cell HepG2 cells using retroviral vector pL-IL-4-SN. Wild HepG2 and IL-4 gene-modified tumor cells were used to induce CTL responses, and cellular surface molecules were evaluated using flow cytometry. Results　The killing activities of tumor cell-specific CTL in IL-4 gene-modified groups were increased from (11.2±2.7)% to (72.5±12)%, which was about seven fold higher than CTL killing activities induced by HepG2 cells (P<0.01). High levels of expression of MHC class Ⅱ antigens as well as the co-stimulatory molecules B7-1 and ICAM-1 were found in IL-4 gene-transferred tumor cells. The expression of MHC class Ⅰwas not affected by IL-4 gene modification. The expression of cell surface molecules induced by IL-4 gene-modified tumor vaccines were completely abrogated through using anti-IL-4 McAb. A significant increase of IL-2 secretions was found during the induction of CTL responses when IL-4 gene-modified tumor cells were used as stimulatory cells. IL-2 secretion and CTL responses were inhibited by anti-IL-4 or anti-surface molecule McAb. Conclusion　IL-4 gene modification enhanced tumor cell-specific CTL killing activities through induction of the expression of cellular surface molecules and facilitation of IL-2 secretion.

Point-of-care testing ( POCT ) is a new area in the laboratory medicine.This article discusses the current situation in regulations , industry management , quality management , personnel management and training and other aspects of POCT in America and China.Then this article points out the challenges of POCT development in China , and further puts forward advanced measures and suggestions for improving POCT management system in China.

Objective To explore the effects of Interleukin 10 (IL 10) on the endothelial damage mediated by inflammatory cytokines in Kawasaki disease (KD).Methods Human umbilical vein endothelial cells were cultured in vitro and the production of tumor necrosis factor alpha (TNF ?),interleukin 1 beta (IL 1?),interleukin 6 (Il 6) and IL 10 were measured by enzyme linked immunosorbent assay (ELISA).The proportion of apoptotic cells in endothelial cells induced by the supernatants of peripheral blood mononuclear cells (PBMCs) was detected by Annexin V/PI double staining though flow cytometry.Electrophoretic mobility shift assay (EMSA) was used to detect the activity of nuclear factor ?B (NF ?B).Results The level of TNF ?,IL 1?,IL 6 and IL 10 were markedly increased in KD patients and the proportion of apoptotic cells in endothelial cells induced by the cultured supernatants of PBMCs was markedly elevated (38 4?7 8)% compared to (2 8?0 8)% in the control subjects.The apoptosis of endothelial cells induced by the cultured supernatants of PBMCs could be reversed to some degree by anti TNF ?,IL 1? and IL 6 monoclonal antibody (McAb),otherwise could be enhanced (58 6?14 6)% by anti IL 10 McAb.The activity of NF ?B in the activated PBMCs in KD patients was distinctly increased and IL 10 could significantly inhibit the activity of NF ?B,reduce the production of TNF ?,IL 1? and IL 6 and the apoptosis of endothelial cells induced by the cultured supenatants of PBMCs in KD patients.Conclusion IL 10,which can inhibit the activity of NF ?B and the transcription of inflammatory cytokines such as TNF ?,IL 1? and IL 6,acts as an important protective factor in endothelial damage of KD.

Objective To study the role of insulin-like growth factor 2 (IGF- Ⅱ ) in the proliferation and degeneration of hemangiomas. Methods The expression of IGF- Ⅱ and Ki-67 was detected with SP im-munohistochemical method in 52 specimens of hemangiomas, 27 of vascular malformation and 5 of normal skin. The expression of IGF-Ⅱ mRNA was examined by reverse transcription polymerase chain reaction (RT-PCR) in specimens from 9 cases of hemangiomas and 5 of vascular malformation. Results The labelling indexes (LI) of IGF- Ⅱ and Ki-67 and the expression of IGF- Ⅱ mRNA were significantly higher in heman gioma than those in vascular malformation and normal skin (P

ObjectiveTo study the alteration mechanism of T follicular helper (Tfh) cells in patients with Kawasaki disease (KD).MethodsTwenty children with KD and the same number of agematched healthy subjects were studied.The proportion of CD4+CXCRS+ICOS+ T in peripheral blood was analyzed by flow cytometry.Real-time PCR was performed to detect the level of Tfh transcriptional factor( Bcl6) and its inhibitor( Blimp-1 ).The plasma concentration of IL-4 and IL-21 were determined by ELISA.ResultsThe proportion of CD4+CXCR5+ICOS+ T in patients with KD was significantly higher than healthy controls [ (2.6±0.6) ％ vs ( 1.8±0.7 ) ％,P＜0.05 ].Transcription levels of Bcl-6 were significantly elevated in patients with KD( P＜0.05),its inhibitor Blimp-1 were found to be down-regulated during acute phase of KD compared with healthy controls (P ＜ 0.05 ).The significant increase of IL-4 and IL-21 plasma concentrations were detected in patients with KD(P＜0.05),in comparison with healthy controls.ConclusionThe over activity of Tfh might be correlated with immune dysregulation in Kawasaki disease.Dysregulation of Bcl-6/Blimp-1,altered microenvironment of IL-4 and IL-21 might be correlated with the abnormal activity of Tfh cells.

ObjectiveTo investigate the role of signal transduction of TLRs in the Henoch-Schonlein purpura (HSP). Methods Reverse-transcription PCR (RT-PCR) and real-time PCR were used to evaluate the levels of TLRs(1～10), MyD88, TRAF6, TRIF, IFN-α, IFN-β, IL-6, IL-1β, TNF-α, IP-10, RANTES,iNOS, Blys/April mRNA expression in peripheral blood mononuclear cells and their expression levels were compared using t test., while the concentration of plasma cytokines such as Blys、IFN-α、IFN-β、IL-6、IL-1、TNF-α was measured by enzyme-linked immunosorbent assay(ELISA).Expression levels of those genes were compared using t test. Results①Compared with the control group, the expression levels of TLR1, TLR2,TLR6, TLR5, TLR3, TLR7, TLR9 mRNA were up-regulated significantly(P＜0.01), while no difference of TLR4 was detected (P＞0.05).②Transcription levels of MyDg8(2.47±1.06) vs(0.73±0.22), TRAF6 (2.54±0.72)×10-3vs(0.70±0.20)×10-3, TRIF(3.18±0.86)×10-3vs(0.93±0.35)×10-3 were significantly up-regulated in acute phase of HSP (P＜0.01).③The levels of IFN-α and IFN-β protein and mRNA were remarkable increased (P＜0.01).④ The expression of cytokine/chemotactic factor such as IL-6, IL-1β, IP-10, RANTES,iNOS was higher than that of the control group(p＜0.01), while TNF-αdid not change in children with HSP (P＞0.05). ⑤ It was detected that the expression of Blys/April was higher than that of the control group(P＜0.01). ConclusionExpressions of TLR1, TLR2, TLR6, TIR5, TLR3, TLR7, TLR9, MyD88, TRAF6, and TRIF are up-regulated during acute phase of HSP, suggesting that aberrant activation of TLRs triggered by microbes may be one of the initiating factors of immune aberrance in HSP. Over expression of cytokine/chemotactic factor or Blys/April owning to the aberrant activation of TLRs, may be correlated with immunological pathogenesis of HSP.

Objective To investigate the effects of the intravenous immunoglobulin (IVIG) on the expression of FcγR Ⅱ b and Toll-like receptor 4 (TLR4) in children with Kawasaki disease (KD).Methods 25 children with KD and 15 healthy controls were enrolled in this study.The levels of TLR4 expression on monocytes were assessed by flow cytometry.Real-time PCR was performed to detect the transcription levels of FcγR Ⅱ b,TLR4 signaling molecules(MyD88,TRAF-6,TAK1) and cytokines(IL-1β,IL-6,TNF-α) in monocytes.The concentrations of TNF-α in plasma was determined by enzyme-linked immunosorbent assay (ELISA).Results (1) Compared with healthy controls,the expression of TLR4 and the signaling molecules in the patients were significantly up-regulated but decreased after treatment with IVIG (P＜0.05).(2)The levels of FcγR Ⅱb expression from the patients were higher than those from healthy controls,and decreased after treatment with IVIG(P＜0.05).(3) The levels of cytokine factor expression such as IL-1β,IL-6 and TNF-α in the patients were found to be higher than those in healthy controls (P＜0.05) ; plasma concentrations of TNF-α were significantly elevated during acute KD (P＜0.05).(4)There were significantly correlations to follow between FcγR Ⅱb and the levels of cytokine,TLR4 expression in monocytes (P ＜0.05).Conclusion IVIG can up regulate FcγR Ⅱb expression and down regulate TLR4 expression which might inhibit the inflammatory response.

Objective To investigate the changes and roles of Fc gamma receptors (FcγR) expressed on monocytes in the immune pathogenesis of Henoch-Schonlein purpura(HSP).Methods Thirty children of HSP and 15 health controls were enrolled in this study.The expressions of FcγR Ⅰ and FcγRⅢ on monocytes were determined by flow cytometry,and real-time PCR was performed to detect the transcription levels of FcγR Ⅱ a,FcγR Ⅱ b,cytokines(IL-1 β,IL-6,TNF-α,IFN-α),chemokine(IP-10,RANTES,iNOS),and BLyS/April in monocytes isolated by microbeads.The plasma concentrations of IL-4,IL-10 and TNF-α were analyzed by enzyme-linked immunosorbent assay (ELISA).Results (1) The expressions of FcγR Ⅰ and FcγR Ⅲ on monocytes in patients with HSP were significantly up-regulated compared with healthy controls.Transcription level of FcγR Ⅱ a on monocytes in patients with acute HSP was found to be higher than that in healthy controls while the inhibitory FcγR Ⅱ] b mRNA was significant down-regulated(P＜0.05),which resulted in a higher ratio of FcR Ⅱ a/Ⅱ b in patients with acute HSP.(2) The expressions of cytokine/chemokines factor such as IL-1 β,IL-6,TNF-α,IP-10,RANTES,and iNOS in patients with HSP was detected to be higher than those in healthy controls(P＜0.05).In addition,expression levels of BLyS/April were up-regulated during acute HSP(P＜0.05),the positive correlations were observed between the FcγR Ⅱ a/FcγR Ⅱ b and the cytokine/chemokines factor in monocytes(P＜0.05).(3) Plasma concentrations of IL-4,IL-10 and TNF-α were significantly elevated during acute HSP(P＜0.05),and a negative correlation was observed between concentrations of TNF-α and the mRNA level of FcγR Ⅱb in monocytes.Conclusion The abnormal expression of the cytokines and the imbalance of stimulatory and inhibitory FcγR of monocyte in acute HSP.

Objective To investigate the effects of sIL-2R in plasma on regulatory T cells (Treg) in children with Kawasaki disease ( KD ) .Methods Thirty-three children with KD and fourteen age-matched healthy children were enrolled in this study .The proportions of CD4+CD25+Foxp3+Treg cells in pe-ripheral blood and mean fluorescence intensity (MFI) of phosphorylated-STAT5 (pSTAT5) protein in CD4+CD25+T cells were analyzed by flow cytometry .The concentrations of sIL-2R, IL-2, IL-7 and IL-15 in plas-ma were measured by cytometric bead array ( CBA ) .Real-time PCR was performed to detect the gene ex-pressions of Foxp3, GITR, CTLA4, IL-2Rα, IL-2Rβand IL-2Rγat mRNA level as well as the expressions of IL-17A and ROR-γt at mRNA level in CD4+CD25-T cells.Results (1) Compared with the control group, the proportions of CD4+CD25+Foxp3+Treg cells in peripheral blood from patients with KD and the expressions of associated factors including Foxp 3, GITR and CTLA-4 at mRNA level were significantly down-regulated (P<0.05).However,the expressions of IL-17A and ROR-γt at mRNA level in Th17 cells were markedly up-regulated (P<0.05), which could be recovered to some extent after treatment with IVIG (P<0.05).(2)The expressions of pSTAT5 protein in CD4+CD25+T cells from patients with acute KD were sig-nificantly decreased (P<00.5 ), but increased with IVIG intervention (P<0.05).(3)The concentrations of sIL-2R in plasma were elevated during acute KD (P<0.05), but decreased after treatment with IVIG (P<0.05).Moreover, KD patients with coronary artery lesion ( KD-CAL+) presented a high level of sIL-2R than those without coronary artery lesion (KD-CAL-) (P<0.05), but there was no significant difference in the concentrations of IL-2, IL-7 and IL-15 in plasma between two groups (P>0.05).(4)The expressions of IL-2Rαand IL-2Rβat mRNA level in CD4+CD25+T cells from patients with acute KD were lower than those of the healthy subjects (P<0.05), but up-regulated to some extent with IVIG treatment (P<0.05).There was no significant change in the expression of IL-2Rγat mRNA level (P>0.05).The concentrations of sIL-2R in plasma were negatively correlated with the expressions of IL-2Rβand Foxp3 at mRNA level and pSTAT5 at protein level (P<0.05).The expression of pSTAT5 protein had positive correlation with the ex-pression of Foxp3 at mRNA level (P<0.05).Conclusion Aberrant IL-2/STAT5 signaling pathway media-ted by significantly increased concentration of sIL-2R in plasma might be one of the factors leading to down-regulation of Treg cells in patients with acute KD .