Objective To investigate the antitumor effect of specific cytotoxic T lymphocytes induced by renal cell carcinoma multiple antigen peptide dendritic cell (DC) vaccine in vitro. Methods Dendritic cells were induced by hGM-CSF,hIL-4 from blood.Peptide of renal cell carcinoma cell line (RCC786-0)was got by citrate-phosphate buffer elution.Peripheral blood mononuclear cell was cultured.Multiple antigen peptide DC cell vaccine was obtained by acid-eluted peptide pulsed DC.The tumor antigen specific CTL was generated from activated T cell by vaccine.Killing activity of the tumor antigen specific CTL was activated by vaccine. Results Antigen peptide DC cell vaccine could show a strong cytotoxic activity of CTLs(31.93±5.05%),which was much higher than control groups(5.88±2.26%,8.03±6.70%,9.70±2.09%,9.35±3.58%). Conclusion Renal cell carcinoma antigen peptide DC vaccine could show a high antitumor effect in vitro.

Objective To study the biological characteristics of the human breast cancer cell mammospheres (MSs) ,and con‐struct breast cancer stem cell experiment model .Methods MCF‐7 cells were cultured in the serum‐free media supplemented with growth factors (the MCF‐7 group) ,and the MSs was collected (the MSs group) .The migration ,invasive and animal tumor forma‐tion abilities of MSs were detected by wound healing ,transwell invasive assay and animal tumor formation test .Results The wound line of MSs healed after 48 hours ,but the line of MCF‐7cells could not heal after 48 h .The number of the cells going through the membrane in MSs group was (76 .24 ± 0 .35) ,and the number in MCF‐7cells was (17 .38 ± 0 .18)(P<0 .05) .MSs had stronger ani‐mal tumor formation ability than MCF‐7 cells .Conclusion MSs have stronger abilities in migration ,invasive and animal tumor for‐mation ,and could be used in the studies of breast cancer stem cell as experimental model .

Objective To study the effect of partial parenterai nutrition support in severe cases of general surgery.Methods Treated 170 severe cases from January 2001 to September 2008,ran two sets, 80 cases of ex-perimental group and 90 cases of the control group.Results The exaltation of hemoglobin, albumin, lymphocytes and weight in experimental group were better than those in the control group (P < 0.05).The level of glycemic control was also better than that in the control group (P < 0.05).Conclusions The effection of partial parenteral nutrition support is reliable, practical and low-cost.

Objective To explore the clinical characteristics,therapy reactions and prognosis of the elderly patients with idiopathic thrombocytopenic purpura(ITP). Methods A total of 43elderly ITP patients(age≥60 years old)including 16 men and 27 women were reviewed and further followed up for 1 month to 15 years. Results Until June 2007,35 elderly ITP patients survived,platelet counts were sustained(30-50)×109/L in 7 cases,but no significant bleeding was found.Thirty-six patients had adrenocorticosteroid therapy first, 25 patients were sensitive to adrenocorticosteroid therapy,4 patients underwent splenectomy,and 3 patients achieved a normal platelet count. Immunosuppressive agents(vinscristine,cyclophosphamide, azathioprine and Cyclosporin A)treatments were held in 5 6 case-times,Cyclosporin A and azathioprine were more effective than vinscristine and cyclophosphamide.The refractory rate was 13.9%.One patient progressed to monoclonal gammopathy of unknown significance and 1 to lymphoma.Eight patients died.1 patient died of brain bleeding after trauma,3 patients died of malignant neoplasm,4 patients died of heart failure induced by infection. Conclusions The clinical features of elderly ITP patients are atypical.the mortal bleeding in them was rare,treatment strategy should be individualized tO each elderly patient.

Objective To investigate the prevalence by age,response to different therapies and outcome in newly diagnosed idiopathic thrombocytopenic purpura(ITP). Methods ITP patients who were hospitalized from July 1992 to December 2006 and followed uD with telephone were retrospectively analyzed.Results 103 patients with ITP were investigated. The time of follow-up was between 2 months to 15years.to adrenocorticosteroid and 4 patients under going splenectomy achieved a normal platelet count. In those immunosuppressive agents:including vincristine,cyclophosphamide,azathioprine and cyclosporin A(CsA)used in the present series. CsA was shown to be more effective. It could increase the platelet count when given together with prednisolone,the effective rate was 81.3%(26/32). Severe side effects like kiney function failure were not found in CsA treated patients so the use of geug in ITP would be recommended.There were 2,1 and 1 ITP patients progressing respectively to Sjogren's syndrome,systemic lupus erythematosus and lymphoma. 7 patients died,1 patient died of cerebral bleeding,2 brain infarction,3 malignant neoplasm and 1 nephrosis The refractory rate of ITP is 17.2%(10/58). Conclusions The morbidity in older people is high. The mortal bleeding in ITP is low. Treatment should be tailored to the individual patient.

Objective Cancer radio-therapy may induce bone damage of the patients.collagen type I gene expressions in osteoblast after radiation indicates the influence of radiation on the function of early and late osteoblast.Methods Bone marrow stromal cells were differentiated into osteoblasts in vitro.and the characteristics was indentified.The collagen type I expressions in early and late stage osteoblasts exposed to 1～4Gy radiation were examined by RT-PCR.Results Compared to control group,collagen type I gene expressions increased in early osteoblast after 1～3 Gy radiation (P＜0.05),while the gene expressions in late osteoblast that cultured 10 days decreased.Collagen type I gene expression in late stage ostoblast after 4 Gy irradiation was greatly higher than that in early stage osteoblast (P＜0.01).Conclusion After 1～3 Gy irradiation,the collagen type I expression in early osteoblast was enhanced,indicating the increased ability of bone formation.The exposure to 1～3 Gy decreased collagen type I expression in late osteoblast and weakened the ability of bone formation.The result of high expression of collagen type I in late osteoblast after 4 Gy irradiation may be the manifestation of compensatory function.

Objective To investigate the effects of pulsed electromagnetic fields (PEMF) on homing and proliferation-related genes of mouse osteoblasts.Methods 9 week-old C57BL/6 mice were treated with PEMF (70 Hz,1 mT) for 4～5 weeks,while mice in control group didn't not receive PEMF.Bone marrow cells of femurs and tibias were flushed out,and the bones were minced and incubated at 37 ℃ with a type Ⅰ collagenase.Bone associated mononuclear cells (MNCs) were isolated via density centrifugation with Lymphoprep.Magnetic cell sorting was used before flow-cytometric sorting,and the ALCAM+Sca-1-cells were collected.The homing and proliferation-related genes expressed in ALCAM+Sca-1-cells were detected with high throughput microarray and RT-PCR.Results The expression of Jag1 and Ang-1 in mouse osteoblasts increased under the effects of PEMF.Conclusions PEMF may have regulation effects on HSC (hematopoietic stem cell) survival through modulating the homing and proliferationrelated genes in ALCAM+Sca-1-osteoblasts.

Objective To investigate the effect of radiation on the expressions of RANKL and OPG in osteoblasts in order to disclose the molecular mechanism of bone injury induced by ionizing radiation.Methods The osteoblasts were differentiated from MC3T3-E1 cells.After 2 or 4 Gy137 Cs γ-irradiation,the mRNA and protein expression levels of RANKL and OPG of osteoblast precursor and osteoblast were detected by real-time PCR and Western blot.Results The expressions of RANKL mRNA (t=5.41,P＜0.05)and protein(t=68.37,P＜0.01)were up-regulated after 4 Gy irradiation,while the expressions of OPG mRNA(t=5.20,7.02,P＜0.05)and protein(t=7.78,9.45,P＜0.05)were down-regulated after 2 and 4 Gy irradiation.Conclusions 2 and 4 Gy ionizing radiation alters RANKL/RANK/OPG pathway in osteoblasts,which may promote the osteoclast differentiation and maturation and hence promote bone resorption of osteoclasts.

Objective To study the influence of irradiation on the osteoblast function by the gene expression changes of RANKL and OPG.Methods Bone marrow stromal cells were induced to develop into early and mature osteoblasts in vitro.The characterization of osteoblasts was indentified by ALP staining.The RANKL and OPG mRNA levels in early and mature osteoblasts, which exposed to 0 -4 Gy radiation were determined by RT-PCR.Results Bone marrow stromal cells had been induced to early and mature osteoblasts by osteoblast differentiation medium in vitro.In early stage of osteoblast, RANKL mRNA expression levels treated with 1Gy irradiation was 2.83-fold higher than those other irradiation dosage groups.The RANKL mRNA expression levels of each group in early stage of osteoblasts were significantly higher than those in the mature counterpart ( t = 8.34 - 103.57, P ＜ 0.05 ).The ratio of RANKL/OPG mRNA was obviously greater in early osteoblast compared with the mature cells ( t = 2.84 - 20.99, P ＜0.05 ), and it was the highest in 1Gy irradiation treated early osteoblast.Conclusions Radiation exposure of the early osteoblasts promotes osteoclasts function and results in the bone loss.

4-Amino dimethyl phthalate as the hapten was coupled to carrier protein and then used to immunize New Zealand rabbits. Polyclonal antibody which showed specific binding to dimethyl phthalate ( DMP) was thus obtained, and on the basis of this, an indirect competitive chemiluminescent enzyme-linked immunoassay ( icCLEIA ) was developed. The experimental parameters of icCLEIA were optimized as follows: the concentration of coating antigen was 50 μg/L, the primary antibody concentration was 92. 5 μg/L, the secondary antibody concentration was 1μg/mL, distilled water (pH 6. 0) was used as diluent solution and the competitive reaction time was 40 min. Under the optimal conditions, the icCLEIA exhibited a linear working range from 0. 74μg/L to 30. 32μg/L with the limit of detection of 0. 29μg/L. The cross-reactivity of thirteen structural analogues was lower than 5%. The recovery of DMP from spiked liquor and soy sauce samples ranged from 80 . 2% to 116 . 0% and the average RSD was less than 3 . 6%. The detection results of the spiked liquor and soy sauce samples were consistent with those by standard gas chromatography-mass spectrometry method. The developed icCLEIA method exhibited a practical potential for detecting DMP residue in food samples.