The study is designed to investigate if human interleukin 18 (hIL-18) cDNA expressed in nasopharyngeal carcinoma (NPC) cells could enhance the cytotoxicity of CD8+ T cells from the PBMC of the MFC-bearing patient and how to enhance it.Methods: A constructed eukaryotic expression vector for human IL-18 ,pcDNA3.1(-)/hIL-18, was used to transfect the NPC cell strain-SUNE cells. Then, the transfected SUNE cells were mix-cultured with CD8+ T cells which were separated from the PBMC of the patient with NPC, the cytotoxicity was measured by lactate dehydrogenase(LDH) releasing method, the expression for Perforin, Fas-L and Granzyme B were detected with the mix-cultures. Results: The eukaryotic expression vector could highly express hIL-18 in transfected SUNE cells and only traces of the protein exist in non-transfected SUNE cells. The concentration of hIL-18 in supernatant of the transfected SUNE cells was (85 ? 10) pg/ml, but less than 5 pg/ml of hIL-18 contained in supernatant of the non-transfected SUNE cells.The expression of hIL-18 in SUNE cells could enhance the cytotoxicity of CD8+ T cells from the NPC patient significantly( P

Objective To establish a solid phase extraction(SPE)-high performance liquid chromatography(HPLC)method for determination of melamine in foods. Methods Melamine in sample was extracted with extractive agent. After centrifugation, the supernatant fluid was purified by Waters Oasis MCX cartridge,and detected with HPLC method. Chromatographic column of TIANHE C18(250 mm?4.6 mm,5 ?m) was used at 35 ℃ and mobile phase was water(containing 0.01 mol/L 1-hexanesulfonic acid sodium salt and 0.01 mol/L citric acid)∶ acetonitrile = 94∶6 (V∶V) at a flow rate of 1.0 ml/min. Injection volume was 20 ?l. Results In the range of 0.80 to 80.0 mg/L, the regression equation was y=84.64 x-6.127 1 with the correlation coefficient of 0.999 98. The detection limit of melamine in foods was 0.5 mg/kg. The average rates of recovery were 88.0%-96.0% and the relative standard deviation was less than 5%. Conclusion The method established in the present paper is applicable to determination of melamine in foods with good precision,accuracy and sensitivity.

Until now, the exactly effect of Kupffer cells (KCs) on inducing immune tolerance or aggravating acute rejection is still unknown. Activated by various way after liver transplantation, have the ability of phagucytosis the apoptositic T cells, up-regulated expression of FasL and many Th2/Th3 cytokines, such as interleukin-10 and transforming growth factor-lB. These up-regulated cytokines could induce the apoptosis of Th1 cells and enhance the proliferation and differentiation of the Th2 cells, finally induce the immune tolerance, However, the activated KCs also have the ability of expression many cytokine-dependent molecules,such as class Ⅱ major histocompatibility antigens, adhesion molecule and costimulatory molecules which could enhance the function of the antigen presentation, increase the expression of Thl cytokine and aggravate the acute rejection after liver transplantation. It maybe relate to the ratio of theTh1/Th2 cells determined by the complicated net of the cytokine produced by the activated Kupffer cells: the predominance of Th2 cells could induce the immune tolerance, on the contrary, the acute rejection proceed.

Objective To study the effects and clinical significance of tea pigment on 24 hour urinary endothelin(UET) excretion in patients with early stage diabetic nephropathy.Methods 65 cases of early stage diabetic nephropathy were randomly divided into expermental group and control group.Control group was treated by routine treatment and the expermental group was treated by tea pigment,in addition to routine treatment,and was orally given tea pigment capsule 0 24g three time daily,for 8 weeks.The amounts of 24 hours UET and urinary albumin excretion rate(UAER)in the two groups patients before and after treatment were measured using radioimmunoassay method.Results The amounts of 24 hour UET in patients with diabetic nephropathy were elevated significantly as compared with those in normal control group(P0 05).After 8 week's treatment,the amounts of 24 hour UET and UAER in expermental group were significantly lower than those of control group(P

Objective To investigate the phosphorylation and dephosphorylation of CDK1 based on the specific cell cycle apoptosis in Molt-4 cells and active variety of CDK1 in cell cycle specific apoptosis.Methods The exponential phase of growth Molt-4 cells (the human acute lymphoblastic leukemia cell line) were induced with dose response and time course of Camptothecin (CPT).The specific cell cycle apoptosis was detected with API method,then cell apoptosis was verified with post sorting confocal method.Meanwhile,the phosphorylation and dephosphorylation of CDK1 were detected by the protein electrophoretic analysis (Western blot).Results The specific cell cycle apoptosis occurred on exponential phase of growth Molt-4 cells after CPT treatment.When Molt-4 cells occured S-phase apoptosis, the apoptosis cell phosphorylation of CDK1-Thr161 band was more narrow than that of control cells, the apoptosis cell phosphorylation of CDK1-Thr15 band almost had the same band with control cells.Conclusion Cell apoptosis frequently developed in different cell cycle phase. API assay is quick and efficient method to analyze specific cell cycle apoptosis. When cell apoptosis take place in S-phase,the phosphorylation activity of CDK1 is inhibited.

Aim To prepare for psoriasis transgenic animals, psoriasis susceptible gene was constructed. Methods Extract gDNA from blood preparations of psoriasis patients, screen positive gDNA owns HLA-Cw*0602 gene, then copy the whole sequence and link it to pMD-19T Simple Vector. Results The identification showed the construction of psoriasis susceptible gene was sucessful. Conclusion The successful construction of HLA-Cw*0602 gene will lay the foundation for development of transgenic animal models of psoriasis.

Objective To compare the efficacy and safety of arsenic trioxide ( ATO) with all-trans retinoic acid ( ATRA) for the treatment of acute promyelocytic leukemia ( APL) . Methods We searched the database of Cochrane Library ( Issue 1,2009) ,CENTRAL ( 1970 to 2009) ,Medline ( 1978 to 2008) ,EMBASE ( 1950 to 2009) ,CBM ( 1978 to 2008) ,CNKI ( 1994 to 2008) and CMAC ( 1994 to 2008) . We also searched the Meta register,Conference Proceedings of American Society of Hematology ( 1946 to 2008) and American Society of Clinical Oncology ( 2004 to 2008) on the internet for grey literature. We had searched the related journals in the library of Third Military Medical University,too. We included randomized controlled trials which compared ATO with ATRA for the treatment of APL. We adopt complete remission rate,overall survival rate, disease-free survival rate,time to complete remission,relapse rate,mortality and adverse reactions as result indicators. Data were entered and analyzed with the Cochrane review manager software ( Revman 5. 0) . Results Four eligible randomized controlled trials ( RCTs) were included ( n =243) . All the RCTs were methodologically graded as B. They all are focusing on the comparison of ATO monotherapy with ATRA monotherapy in treating newly diagnosed APL patients. Meta analysis showed that effect index for complete remission,2-year disease-free survival,time to complete remission,relapse rate and mortality was 0. 96 ( 0. 50,1. 86) ,2. 76 ( 0.71,10.66) ,-1.30 d ( -1.83,-0.78) ,0.86 ( 0.45,1.63) ,and 1.15 ( 0.45,2.95) ,respectively. All indicated no statistically significant difference. Effect index for incidence of liver dysfunction was 3. 03 ( 1. 25, 7. 37) ,which showed statistically significant difference between ATO group and ATRA group. Conclusion ATO is not superior to ATRA in treating newly diagnosed APL patients regarding complete remission,diseasefree survival rate,time to complete remission,relapse rate and mortality. What is worse,it will increase the incidence of liver dysfunction during treatment. Due to limitation of included trials,this conclusion need to be validated by further studies.

Objective To investigate the effect of early enteral nutrition on prognosis of severe acute pancreatitis (SAP) .Methods Fifty-eight patients with SAP were randomly divided into early enteral nutrition group (experimental group ,n=30) and total par-enteral nutrition(TPN) group(control group ,n=28) .The experimental group was feed by Nose-jejunum nutrition tube and the con-trol group were supported with TPN through central vena .Compared the differences in complication incidence rate ,infection rate , mortality rate ,length of hospital stay and costs between two groups .Results The incidence rate of complications ,infection rate , length of hospital stay and hospital costs in experimental group were lower than control group ,the difference was statistically sig-nificant(P<0 .05) .The mortality in experimental group was lower than control group ,but the difference was not statistically sig-nificant(P>0 .05) .Conclusion Early enteral nutrition support therapy can improve the nutritional status of patients with SAP ,re-duce the incidence of complications ,infection ,length of hospital stay and hospital costs .

Objective To investigate apoptosis of gastric cancer cells induced by taxol,and its specific apoptotic cell cycle phase. Methods The apoptosis rate of gastric cancer cell line MKN-28 induced by taxol was detected with Sub-G1 method.The specific apoptotic cell cycle phase Was detected with API and PSC method.The sensitivity of 20 cases clinic gastric cancer specimens induced by taxol was detected with the method MTT.Results With the Sub-G1 method the MKN-28 cells were induced by 10 μg/ml taxol,after 10 h,the apoptosis rate reached its top apex;with the API method and the PSC method,the specific apoptosts took place in G_2/M phase;the chemotherapy sensitivity of 16 cases out of 20 cases clinic gastnc cancer specimens exceed 50%with the method MTT. Conclusion Taxol could induce gastric cancer cells to aimptosis and the apoptosis takes place in G_2/M phase;Taxol is sensitive to clinic gastric cancer speclmens.

In order to evaluate the effect of mitofusin-2 gene (mfn2) on proliferation and chemotherapy sensitivity of human breast carcinoma cell line MCF-7 in vitro, pEGFPmfn2 plasmid carrying full length of mitofusin-2 gene was transfected, by using sofast, into MCF-7 cells. Mitofusin-2 gene expression in MCF-7 cells transfected by sofast after 48 h was detected by PCR and Western blotting, and the stable expression of GFP protein in MCF-7 cells by Western blot analysis. The proliferation of MCF-7 cells was assayed by MTT and cell counting. By using PI method, the effects of mfn2 on the cell cycle distribution of MCF-7 were measured. Annexin-V/PI double labeling method was employed to detect the changes in apoptosis induced by chemotherapeutics before and after transfection. The results showed that the MCF-7 cells transfected with mfn2 gene could stably and highly express GFP protein. MTT assay revealed that after transfection of mfn2 cDNA, the proliferation of MCF-7 cells was significantly inhibited. DNA histogram showed that cells arrested in S phase, and the percentage of S phase cells was 42.7, 17.2 and 19.6 in mfn2 cDNA transfection group, blank plasmid transfection group and blank control group, respectively (P<0.05). The apoptosis ratio of the cells transfected with mfn2 gene was increased from 3.56% to 15.95%, that of the cells treated with camptothecin (CAMP) followed by mfn2 gene transfection was 69.6%, and that in blank plasmid transfection group and blank control group was 31.0% and 23.4% respectively (P<0.05). It was suggested that transfection of mfn2 gene could significantly inhibit the proliferation of MCF-7 cells and promote their sensitivity to CAMP with a synergic effect.
Antineoplastic Agents, Phytogenic/pharmacology ; Apoptosis ; Camptothecin/pharmacology ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Drug Screening Assays, Antitumor ; Flow Cytometry ; Gene Expression Regulation, Neoplastic ; Green Fluorescent Proteins/metabolism ; Membrane Proteins/*biosynthesis ; Membrane Proteins/*genetics ; Mitochondrial Proteins/*biosynthesis ; Mitochondrial Proteins/*genetics ; Transfection