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 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.

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.

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 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 role of T follicular helper ( Tfh) cells in the immuno-pathogenesis of persistent immune thrombocytopenic purpura ( pITP) .Methods Twenty children with pITP and twenty healthy controls were enrolled in this study .The proportion of CD4+CXCR5+ICOShigh PD-1high T ( cTfh) cells and the expression of ICOSL on CD 19+B cells in peripheral blood of the patients and healthy subjects were analyzed by flow cytometry .The expressions of Bcl-6, c-Maf, IL-21 and ICOSL at mRNA level were detected by real-time PCR.The plasma concentrations of IL-2, IL-6 and IL-21 were determined by ELISA.Results (1) Compared with the healthy controls , the proportions of cTfh cells increased signifi-cantly in patients with pITP [(17.45±9.04) %vs.(7.57±2.57) %, P<0.05], but decreased with the treatment of dexamethasone (DEX) for 7 days [(5.93±1.64) %vs.(17.45±9.04) %, P<0.05].(2) The expression of Bcl-6, c-Maf and IL-21 at mRNA level in patients with pITP were higher than those in healthy controls (P<0.05).(3) Compared with healthy controls, the expression of ICOSL at mRNA and protein levels in CD19+B cells were significantly up-regulated in patients with pITP (P<0.05), which showed no significant changes after treatment with DEX (P>0.05).(4) The plasma concentration of IL-21 was remarkably elevated in patients with pITP , regardless of DEX treatment (P<0.05).But the plasma concentrations of IL-2 and IL-6 showed no significant changes compared with control group (P>0.05).Con-clusion The immunopathogenesis of persistent immune thrombocytopenic purpura might be associated with the hyper-activation of Tfh cells caused by excessive expression of ICOSL and IL-21.The persistent high expression of ICOSL and IL-21 might be one of the important factors resulting in the recurrence of pITP in children .