Objective To study proliferation,secreted cytokines,immune phenotypes and cytotoxicity on Raji cells by cytokine induced killer (CIK) cells co-cultured with dendritic cells (DC).Methods The mononuclear cells from peripheral blood of healthy individuals were extracted,then cultured the cells under 5 ％ CO2 at 37 ℃ for 2 hours.DC were induced from suspended cells,and CIK cells were from adherent cells.After 9 days of nurturing,two types of cells were mixed.CIK cells were cultured alone as the control.The cytotoxic activity of CIK and DC-CIK cells were detected by MTT assay.The morphologies,proliferation,secreted cytokines,and immune phenotypes of the two cells in day 0,3,6,9,12,15 in culture were monitored.Results In day 12 in culture,comparing with CIK cells,DC-CIK cells significantly enhanced the proliferation rate [(42.44±2.68) fold vs (30.01±2.05) fold] (t =11.64,P ＜ 0.05) and had increased IL-2,IFN-γ,IL-12 and TNF-α secretion [(124.34±12.57) ng/L vs (56.32±6.58) ng/L,(496.60±95.32) ng/L vs (247.80± 69.45) ng/L,(84.92±6.07) ng/L vs (24.18±3.31) ng/L,(380.6±45.95) ng/L vs (196.61±24.19) ng/L] (t =15.16,P ＜ 0.05; t =6.67,P ＜ 0.05; t =27.78,P ＜ 0.05; t =11.20,P ＜ 0.05),and there were more CD3+ CD8+ cells and CD3+ CD56+ cells in the co-culture [(71.79±1.73) ％ vs (60.37±3.24) ％,(48.54±3.30) ％ vs (33.07±2.22) ％](t =9.83,P ＜ 0.05; t =12.30,P ＜ 0.05),and DC-CIK cells had a significandy increased cytotoxicity on Raji cells in vitro at the same ratio of effector cells to target cells.Conclusion CIK cells have higher proliferation rate and cytotoxicity against Raji cells when co-cultured with DC.

0.05).Conclusion CAP infiltration at trigeminal nerve infraorbital branch region in rats can produce stronger analgesic effect,it related to the dosage,concentration,using dosage, time and intermission.

The absorption process is an important factor in determining the bioavailability of orally administered drugs. however, the absorption mechanism of many drugs is not clear. Caco-2 cell model is the best in vitro absorption model nowadays. It is widely used in the research on drug absorption process and absorption mechanism, especially in the aspect of traditional Chinese medicine absorption, the use of Caco-2 cell model has become the hot spot recently. Mo- reover, Caco-2 cell model is also applied in the research on drug metabolism. Therefore, Caco-2 cell model will become an important method in the research of drug absorption, and will be helpful to accelerate the process of new drug screening and development.

Objective To investigate the differential gene expression in chronic hepatitis B (CHB) patients with typical syndromes of traditional Chinese medicine ( TCM) syndromes, and to explore the relationship between TCM syndromes and gene expression. Methods Peripheral blood samples were collected from CHB patients and healthy volunteers before treatment. After total RNA of leukocytes was isolated, the gene expression profiles were detected by microarray. The expression levels of partial genes were tested by real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR) . Results Microarray analysis results showed that there were significant differences of gene expression between CHB patients and healthy volunteers, and among CHB patients with the syndrome of liver-qi stagnation and spleen deficiency, syndrome of damp-heat accumulation, and syndrome of liver-kidney yin deficiency. The results of gene ontology ( GO) and signal pathway analysis between the healthy control and CHB patients with various syndromes showed that the specially-regulated genes of CHB patients with liver-qi stagnation and spleen deficiency were mainly related to cytokinetic process, those in patients with dampness-heat accumulation were mainly related to the positive regulation of lipid storage, and those of patients with liver-kidney yin deficiency were mainly related to the activities of nitric oxide synthase regulator. The real time RT-PCR for partial genes presented the similar results with those of gene microarray. Conclusion There are specific expression profiles of differential genes and significant differential genes in CHB patients with the syndrome of liver-qi stagnation and spleen deficiency, syndrome of damp-heat accumulation, and syndrome of liver-kidney yin deficiency, which may be the molecular foundation for the classification of TCM syndromes of CHB patients.

Objective To explore the effects of positive end-expiratory pressure on intraoperative pulmonary function and respiratory mechanics in patients receiving continuous hyperthermic peritoneal perfusion.Methods Ninety patients (55 males,35 females,aged 40-70 years,ASA grade Ⅰ-Ⅲ) undergoing continuous hyperthermic peritoneal perfusion were selected and divided into 3 groups (n=30 each): regular volume controlled ventilation group (group A),5 cm H2O PEEP group (group B) and 10 cm H2O PEEP group (group C).After tracheal intubation,the mechanical ventilation parameters in groups A,B and C were respectively given tidal volume (VT) 10 ml/kg without positive end-expiratory pressure (PEEP),VT 6 ml/kg with 5 cm H2O PEEP,and VT 6 ml/kg with 10cm H2O.PETCO2 was maintained at 35-45 mm Hg.Arterial blood samples were collected for blood gas analysis 5 min after (T1),before CHPP (T2),the end of CHPP (T3),and before the end of mechanical ventilation (T4).Besides,Pplat,Pmean,Ppeak,PaCO2,PaO2were recorded and Cdyn,OI,RI,A-aDO2 as well as VD/VT were calculated at all time points simultaneously.Pulmonary complications during 7 days after surgery were also recorded.Results Compared with group A,Ppeak,Pplat,A-aDO2 and RI were all significantly lower (P<0.05),while OI and VD/VT were higher in groups B and C at T1-T4 (P<0.05);at T2-T4,Cdyn and PaO2 were higher with lower Pmean in groups B and C (P<0.05).Compared with T1,Ppeak,Pplat and Pmean were higher (P<0.05) while Cdyn was lower (P<0.05) in group A at T2-T4;In Group B,Ppeak and Pplat were higher at T3 (P<0.05),Pmean was higher at T2-T4 (P<0.05) and Cdyn was lower at T3,T4 (P<0.05);in group C,Ppeak,Pplat and Pmean were all higher at T2-T4 (P<0.05),Cdyn was lower at T3,T4 (P<0.05);OI and PaO2 were lower (P<0.05),while A-aDO2,VD/VT and RI were all higher (P<0.05) at T2-T4 in the three groups.In addition,the incidence rates of pulmonary infection,hypoxemia,and atelectasis were significantly lower in groups B and C during 7 days after surgery than those of group A (P<0.05).Conclusion PEEP (5 cm H2O) with VT (6 ml/kg) could effectively improve intraoperative pulmonary function and reduce the risk of perioperative pulmonary complications of the patients receiving continuous hyperthermic peritoneal perfusion.

Objective To investigate the expression level of lipocalin-type prostaglandin D synthase (L-PGDS) mRNA and its protein in glioma.Methods The L-PGDS mRNA was determined by real-time quantitative RT-PCR in 23 glioma and 5 healthy brain specimens. L-PGDS protein was detected by Western blot in cerebrospinal fluid (CSF) and brain tissue specimens.Results In 23 glioma specimens L-PGDS mRNA expression ranges from 1.5?10 3～7.0?10 4 copy/?g RNA, mean value is 1.6?10 4 copy/?g RNA. In 5 normal brain specimens, the mean value is 8.1?10 5 copy/?g RNA, statistic analysis indicates a distinct discrepancy ( P

Objective To evaluate the effects of Radix Astr agali combined with prednisone and i mmunosuppressant for primary nephrotic syndrome (PNS)in adults and to compare the effects o f Radix Astragali in various prepata tions for PNS.Methods Randomized controlled trials were a pplied for systemic reviews.Electr onic and manual retrieve of Medline,Embase,Cochrane Library,CBMdisc a nd CEBM/CCD and relevant medical jou rnals in China were applied to search the RCTs of Radix Astragali,non -specific treatment,glucocorticoids and i mmunosuppresants for PNS,and the RCTs were analysed with RevMan 4.1.Results There were 14randomized controlled trials with 524cases involved.Meta-analysis showed that Radix Astragali could in crease the therapeutic effect of pre dnisone and immunosuppressant for PNS and re-duce its recurrence.Radix Astragali also had an effect in decreasing 24-hour proteinuria content and the pla sma levels of total cholesterol and albumin.There were no differences between single injection and compound decoction.Asymmetry showed in"Funnel plot"may be related to publication bias,l ow quality of methodology and small -size in sample.Conclusion Radix Astragali and its prescriptio n may become a prospect therapy for PN S and its recurrence and the com-bination of traditional Chinese med icine and western medicine can be more effective for PNS.The dedinite effect of Radix Astragali for PNS will be further con firmed by multiple -center,large -s ample randomized controlled trial.

Objective To observe the change of serum ischemia modified albumin (IMA) in continuous ambulatory peritoneal dialysis (CAPD) patients.Methods Seventy-four end stage renal disease patients undergoing CAPD more than 3 months (CAPD group) were divided into 2 groups according to clinical symptoms of cardiovascular diseases:CAPD symptoms group (29 cases) and CAPD asymptomatic group (45 cases).Seventy healthy subjects were selected as control group.The serum IMA level and abnormal rate of electrocardiogram and heart color ultrasonic were examined and compared.Results The serum IMA level in CAPD group was significantly higher than that in control group [(92.33 ± 17.17) kU/L vs.(69.63 ± 9.24)kU/L,P＜ 0.01].The serum IMA level in CAPD symptoms group was significantly higher than that in CAPD asymptomatic group [(109.37 ± 21.34) kU/L vs.(85.31 ± 8.58) kU/L,P ＜ 0.05].The elevatory rate of serum IMA level and abnormal rate of electrocardiogram and heart color ultrasonic in CAPD symptoms group were significantly higher than those in C APD asymptomatic group [37.9％ (11/29) vs.15.6％(7/45),62.1％ (18/29) vs.22.2％ (10/45),P ＜0.05].Conclusions The serum IMA level in CAPD patients is elevatory.Serum IMA level is significantly higher in CAPD patients with cardiovascular disease clinical symptoms,it can be used for diagnosis of myocardial damage in CAPD patients.

Objective To study the killing effects of the killer cell immunoglobulin-like receptors (KIR)2DS1-mediated natural killer (NK) cells against dendritic cells (DCs) and analysis the expression of chemokine receptor CCR7 after KIR2DS1-positive NK cells interact with the C2 epitope DCs in vitro,and explore the mechanism of alloreactive NK cell to limit graft-versus-host disease (GVHD).Methods NK cells were separated by Rosettesep NK sorting kit from healthy donor peripheral blood and taken as effector cells.The mononuclear cells from peripheral blood of newly diagnosed acute myeloid leukemia (AML) patients,and DCs were induced as target cells.Polymerase chain reaction and sequence specific primers (PCR-SSP) genotyping techniques and direct sequencing SBT were used to detect KIR gene,HLA-Cw of the healthy donors and patients.The cytotoxic activity of KIR2DS1-positive NK cells or KIR2DS1-negative NK cells against DCs were detected by CCK-8 assay.The expression of CCR7 after NK cells interacted with DCs was detected by flow cytometry.Results The purity of NK cells analyzed by flow cytometry was (94.20±1.23) ％.Under same target ratio (10∶1),the killing effect of KIR2DS1+ NK cell against DCs was higher than that in KIR2DS1-NK cells (t =3.70,P ＜ 0.05).Under same target ratio (10∶1),the cell lysis in C1/C1 group,KIR2DS1+ NK cells against C2/C2 group DCs (15.06±1.81) ％ was significantly higher than that against C1/C1 group (10.07±1.03) ％ and C1/C2 group (8.65±0.93) ％ DCs (F =56.368,P =0.001).KIR2DS1 + NK cells efficiently acquired CCR7 upon coculture with C2 epitope DCs.Conclusions NK cell expressing the KIR2DS1-activating receptor efficiently kill DCs,prevent GVHD,while enable NK cell acquire CCR7,obtaining migratory properties in vitro.

BACKGROUND:Mesenchymal stem cells are found to have the immunoregulatory activities and a potential application prospect in the treatment of autoimmune diseases.
OBJECTIVE:To explore the mechanism of transplanting mesenchymal stems cells on the treatment of multiple sclerosis.
METHODS:The mouse mesenchymal stems cells were prepared, and injected into the al ogenic and syngenic normal mice, to detect the frequency of CD4+CD25+Foxp3+T cells in the spleen, thymus, and lymph nodes by flow cytometry, and to detect the Foxp3, transforming growth factor-β1, and interleukin-10 mRNA in the spleen, thymus, and lymph nodes by reverse transcription-PCR.
RESULTS AND CONCLUSION:Transplantation of mesenchymal stem cells on normal mice led to a significant up-regulation of CD4+CD25+Foxp3+T cells, Foxp3, transforming growth factor-β1, and interleukin-10 mRNA in the spleen, thymus, and lymph nodes both in the al ogenic and syngenic transplant groups. Transplantation of mesenchymal stem cells may be an available method in the treatment of autoimmune diseases, and CD4+CD25+Foxp3+T cell, Foxp3, transforming growth factor-β1, and interleukin-10 may be involved in this process.