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.

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.

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.

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.

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 establish a method for indentification of the podophyllum emodi species, Radix Clematis and Radix Gentiana species. Methods: HPLC/UV fingerprint analysis method of toxic ingredient podophyllotoxin and its derivatives were developed, and the method had been evaluated. Results: The methodological evaluation showed that this method had a good repeatability and reproducibility, and different samples had different HPLC fingerprints. Conclusion: This method can be used to differentiate podophyllum emodi from two commonly used medicinal herbs of a different genus but having similar appearance, Radix Clematis and Radix Gentiana.

Objective To obtain the Tat-GFP fusion proteins with penetrating activity and labeled with green fluorescence protein (GFP), and to explore the cell membrane penetrating activity of Tat-GFP in MCF-7 cells. Methods The plasmid pET-24a-Tat-GFP was transformed into Escherichia coli BL21 cells. Different concentrations (0.5 and 1.0 mmol · L-1 ) of isopropyl-β-D-thiogalactopyranoside (IPTG ) and cell culture temperatures (22℃ and 37℃)were used to optimize the protein expression.The Tat-GFP proteins in supernatant were purified using Ni-IDA resins. Western blotting analysis was used to identify the Tat-GFP protein, and confocal laser scanning microscope (CLSM ) was used to examine the cell penetration of Tat-GFP protein. Results There was no significant difference in the Tat-GFP protein production induced by 0.5 and 1.0 mmol·L-1 IPTG;however,the low temperature (22℃)-induced BL21 cells expressed more Tat-GFP proteins than that at 37℃ induction.The Western blotting analysis results showed that GFP antibody could specifically recognize the proteins in PVDF membranes in dose-dependent manner;the CLSM results indicated the distribution of green fluorescence in cytoplasm and nucleus of MCF-7 cells.Conclusion The Tat-GFP protein highly expresses in the supenatant of Escherichia coli i BL2 1 cells at low temperature;the obtained Tat-GFP protein with green fluorescence preserves the cell penetrating activity.

Objective To analyze the airflow characteristics and investigate the relationship of the structure and the function of nasal cavity by the three?dimensional reconstruction of nasal airway of patients with structural abnormalities(nasal septum deviation)and healthy people and the estab?lishment of finite element model by computer. Methods On the basis of CT imaging of the nasal cavity in patients with structural abnormalities(na?sal septum deviation,n=20)and healthy people(n=20),three?dimensional reconstruction of nasal airway was conducted by resurfacing finite ele?ment subdivision to simulate the characteristics of airflow in nasal cavity. Results The airflow mainly went through the commodious side of the nose in patients with nasal septum deviation and the maximum fluence appeared in the middle part of meatus nasi communis. The airway pressure de?creased most significantly in the most flank?curvature part of nasal septum deviation,accounting approximately 79.65%of the total pressure. In healthy people,the bilateral airflow was affected by nasal cycle and was mainly characterized by one nasal cavity,and the maximum fluence was ob?served in the middle and the inferior part of meatus nasi communis. The airway pressure decreased most significantly in limen nasi,accounting ap?proximately 58.78%of the total pressure. Conclusion Numerical modeling of nasal cavity can be used to analyze the relationship between the nasal structural abnormalities and the airflow characteristics,which is a scientific method to analyze the association of nasal structure and function with dis?ease and can be used for pre?and post?operative individual evaluation of operative therapeutic regimen targeting at optimizing airway and altering air?flow distribution.

Objective To study the killing effects of the killer cell immunoglobulin-like receptors (KIR) KIR2DS1-positive natural killer (NK) cells against leukemia cells.Methods High-purified NK cells separated by RosetteSep NK cell enrichment kit from healthy donor peripheral blood were taken as effector cells,and the freshly isolated bone marrow mononuclear cells from newly diagnosed acute myelogeneous leukemia (AML) patients were taken as target cells.The cytotoxic activity of NK cells were detected by CCK8 kit assay.HLA-Cw,KIR gene of the healthy donors and patients were detected by polymerase chain reaction and sequence specific primer (PCR-SSP) genotyping techniques,respectively.NK cells were divided into KIR2DS1-positive group and KIR2DS1-negevitive group,and then anti-KIR2DS1 mononuclear antibody was used to block KIR2DS1 of NK cells.Meanwhile based on HLA-Cw,KIR2DS1-positive NK cells and target cells were divided into C1 homozygote group(expressing HLA-Cw 01,03,07,08,12,14,16 alleles),C2 homozygote group (expressing HLA-Cw 02,04,05,06,15,17,18 alleles) and the C1/C2 heterozygote group (co-expressing the alleles in C1 group and C2 group).Results The purity of NK cells was (91.2±5.94) ％ through flow cytometry analysis.At the same effector-target ratio,cytotoxicity of KIR2DS1-positive NK cells against target cells was higher than that of KIR2DS1-negetive NK cells.While the E:T =10:1,cytotoxicity of KIR2DS1-positive NK cells against target cells [(2.82±6.81) ％] was significant higher than that of KIR2DS1-negetive NK cells [(28.61±5.14) ％] against target cells.The killing effects of KIR2DS1-positive NK cells was significantly weakened after KIR2DS1 blockaded with specific antibody (t =-3.00,P =0.05).When focusing on the C1 group NK cells,KIR2DS1-positive NK cells against C2 group [(4.39±3.46) ％] target cells was significantly higher than than against C1 group [(41.22±3.68) ％] (t =8.33,P ＜ 0.05) and C1/C2 group [(41.32±5.09) ％] (t =6.37,P ＜ 0.05) target cells.Conclusions The killing effects of KIR2DS1-positive NK cells are significantly higher than that of KIR2DS1-negetive NK cells.The HLA-C1 group KIR2DS1-positive NK cells could recognize HLA-C2 loci and then kill the target cells.