Objective To investigate the induction of specific anti-tumor immune response by transfected dendritic cells (DCs) with survivin mRNA of human pancreatic cancer, and to provide the experimental evidences for the treatment of human pancreatic cancer with DCs vaccine. Methods DCs were isolated and cultured from peripheral blood mononuclear cells (PBMCs). After being transcripted and amplified, survivin mRNA was transfected into DCs by electroporation. The expression of survivin in DCs at different time points was detected by quantitative real-time PCR. The survival rate of DCs before and after transfection was determined by MTT method. The induction of specific cytotoxic T lymphocyte (CTL) response by survivin mRNA transfected DCs was measured by 51Cr standard cytotoxicity test. The induction of specific CTL activation by survivin mRNA transfected DCs was evaluated through testing released IFN-γ by ELISA method. Results After survivin mRNA transfection for 48h, the expression of survivin mRNA in DCs reached the highest point (46.09±6.57). After transfection, the survival rate of DCs was stabilized around 80%. The DCs transfected with survivin mRNA could effectively induce HLA-A2+ / survivin+ specific CTL immune responses. Stimulated with pancreatic cancer cell line Capan-2 cells or SCL-1 cells as control group, the IFN-γ released in 24 hours by survivin specific CTL were (28.79±5.70) U/ml and (25.12±2.13) U/ml respectively, there was no significant difference (P=0.761). Conclusion The induction of CTLs by DCs transfected with human pancreatic cancer survivin mRNA could produce specific anti-tumor immunity.

Objective To investigate the best method of transfecting total RNA extracted from pancreatic cancer MiaPaCa-2 cells into dendrtic cells (DCs). Methods DCs were cultured from peripheral blood mononuclear cells induced by rhGM-CSF, rhIL-4 and TNF-α. Morphology of DCs was observed. Flow cytometry was used to detect the mature DCs specific surface markers:CD40, HLA-DR, CD83, CD86. Mixed lymphocyte (MLR) was used to determine the ability of DCs to stimulate allogeneic T cell proliferation.Liposomal transfection, electroporation method and passive transfection was used to transfect MiaPaCa-2 cell total RNA into DCs, Real time RT-PCR and MTT assay was used to determine the expression of MUC1 mRNA and the survival rate of the RNA transfected DCs. Results The cells acquired showed typical DCs morphology, the positive rate of CD40, HLA DR, CD83 and CD86 were 34.3% ,50.2% ,89.2% and 73.6%,and they showed a strong ability to stimulate allogeneic T cell proliferation. 48 h after transfection with MiaPaCa-2 cells total RNA by using electroporation, the MUC1 mRNA amount (45.39 ± 9.33) in DCs was higher than that of liposomes method (3 1. 68 ± 7.25) and passive transfection method (18.53 ± 3.26) . DCs survival rate was (80.36 ± 2.43)% by using electroporation, which was relatively lower than (91.48 ±5.42) % by using passive transfection method, but higher than (67.44 ± 2.51) % by using liposomes method,and it was stabilized around 80%. Conclusions Transfecting total RNA extracted from pancreatic cancer MiaPaCa-2 cells into DCs with electroporation is efficient and safe.

PURPOSETo evaluate the radiosensitizing effect of cisplatin combined with radiotherapy for esophageal carcinoma. METHODS From 1989 to 1990. 60 patients with esophageal were treated. It was divided into two groups. Radiotherapy group (R) and radiotherapy plus cisplatin (20mg iv. twice a week) (R + C). A total dose of 66-70 Gy/6-7 wk was used in (R) and (R+C) group. RESULTS The 1 year. 3 year. 5 year survival rats were R : R+C = 46. 7%、20%、10% : 60%、30%、23%. CONCLUSION This study may imply that rational administration of cisplatin in the radiotherapy could improve the radiotherapy result of the esophageal carcinoma.

The tongue and its surrounding structures have special anatomical and functional characters and how to restore its shape and function after glossectomy (especially total glossectomy) still remains a difficult problem. The authors devised a new method of repair by using a forearm free flap with results in 16 out of 17 patients. The selection and shaping of the flap, the evaluation of the recipient site and its blood vessels, the prevention of the oral infection,the monitoring of the circulatory status of the flap and the evalution of the operative result were discussed.

Objective: To summarize the experience of repairing or al and maxillofacial defects with radial forearm free flap. Methods: The clinical records of 110 cases received oral and maxillofacial reconstruction with radial forearm free flap were analyzed.Results: Forearm free flap healed well in 10 5 out of the 110 cases.with the overall success rate of 96.4%.Failure was in 4 c ases of cancer.One patient died from cerebral infarction 19 days after operation .Post operation infection did not result in the necrosis of the flaps because o f corresponding treatment in 5 cases.Conclusion: The radia l forearm free flap characterized by the anatomical consistency,easy fold, ideal thickness, flexibility and long pedicle, is feasible in the reconstruction of soft tissue defects in oral and maxillofacial region.

Objective To evaluate the clinical results of late course accelerated hyperfractionation(LCAF) radiotherapy for nasopharyngeal carcinoma(NPC) patients . Methods From October 1995 to September 1996, 136 NPC patients were divided into conventional fractionation(CF) and LCAF radiotherapy groups.Seventy patients in CF group received conventional radiotherapy :2.0 Gy/f,5 fractions a week to a total dose of 70 Gy in 47 49 days .Sixty six patients in LCAF group were treated with the same fractionation as CF group untill the dose of 36 40 Gy ,and then followed by LCAF radiotherapy :1.35 Gy/f ,twice daily with 6 hour interval between the two fractions,to the total dose of 75.1 76.5 Gy in 45 47 day .Results The complete response rate in the nasopharynx was 97% in the LCAF group and 90% in the CF group after 3 months of radiotherapy.The 1 ,3 ,and 5 year local control rates were 97.0%, 95.4%,89.9% in the LCAF group and 92.8%,77.1%,70.1% in the CF group ,with the difference between the two groups significant (P0.05 ),without finding any significant difference in acute mucusitis or late complications between these two groups. Conclusions The local control rate of NPC treated by LCAF radiotherapy is superior to CF. However there is no significant difference in the survival rates between the two groups .LCAF radiotherapy is well tolerated and not increase the toxicity.

0.05).The scores of MMSE increased significantly(all P

Objective:To develop a scale used to measure smoking-related attitudes in Chinese secondary school students. Methods:Based on literatures published in China and abroad and suggestions by relevant specialists and teachers, we developed a self-administered scale with 16-item used to measure smoking-related attitudes in Chinese secondary school students. Factor analysis was used to assess the construct validity of the scale. The differences of the scores between trying smokers and non-smoker, and males and females were tested for assessing the discriminant validity of the scale. Cronbach's alpha for the total scale and the sub-scales was calculated for evaluating the internal consistency, and the split-half reliability test analysis was also conducted. Moreover, 112 subjects were re-investigated after two weeks of the first survey, and the difference and correlation of scores between the two surveys were tested for assessing the stability of the scale. Results: Factor analysis identified two potential components that could explain 46.2% of the total variance, and the first factor including 9 items was defined as "The opinions to tobacco and smoking-related behaviors", and the second one including 7 items was defined as "The opinions to measures for preventing and controlling smoking". The mean score of the scale in non-smokers was significantly higher than that of the trying smokers, while the mean score in females was significantly higher than that in males. The Cronbach'? coefficient was 0.87 for Factor One, 0.75 for Factor Two, and 0.86 for the general scale, and the coefficient of the split-half reliability was 0.71 for Factor One, 0.59 for Factor Two, and 0.72 for the general scale. By analyzing the data of 112 students who participated in both surveys, we obtained test-retest reliability of factorⅠas 0.72, of factor Ⅱ as 0.44 and of all items as 0.67, and all the correlation coefficients were statistically significant. There were no differences for the scores of the scale between the two surveys. Conclusion: The results indicated that the smoking-related attitude scale had reasonable validity and reliability,It could provide valuable reference for future similar surveys in China.

Objective:To study the effects of rat interleukin-10 (rIL-10) gene treatment on the expression of collagen , matrix metalloproteinase 13(MMP13) and their specific inhibitors the tissue inhibitor of metalloproteinase 1(TIMP1) in porcine serum in-duced liver fibrosis rats then to explore the anti-fibrotic effect of rL-10.Methods:Thirty SD rats were divided into normal control and fibrosis model group.Normal control group (group C) was intraperitoneally injected with 0.5 ml normal sodium twice a week for 8 week, while the fibrosis model group was injected with equal volume of pig serum for 8 week.At the beginning of the 5th week, fibrosis model group was further randomly divided into a fibrosis model subgroup ( group M ) , rIL-10 gene treatment subgroup ( group I ) and empty vector control subgroup(group P).Rats in group C and M were injected with Ringer’s solution as a reagent control via the tail vein weekly, rats in group I were injected with the rIL-10 plasmid pcDNA3-rIL-10, and rats in group P were injected with empty vector pcDNA3.All rats were sacrificed at the end of 8th week, and the liver tissue samples were collected to observe deposition of collegan in liver tissue by sirius red staining and detected the expression of MMP 13 and TIMP1 in the liver tissue by SP immunohistochemistry .Re-sults:Sirius red staining showed that the area of the collegan deposition was dramatically increased in fibrosis model subgroup and emp -ty vector control subgroup compared with the normal control group , and the area of the collagen deposition was dramatically decreased in rIL-10 gene treatment subgroup compared with the fibrosis model and empty vector control subgroup .Immunohistochemistry analysis showed that the expression of MMP 13 and TIMP1 in fibrosis model subgroup and empty vector control subgroup was significantly higher than the normal control group , but compared with normal control group , expression of MMP13 was significantly increased and expres-sion of TIMP1 was significantly decreased in rIL-10 gene treatment subgroup .Compared with fibrosis model subgroup and empty vector control subgroup, the expression of MMP13 and TIMP1 was dramatically decreased in rIL-10 gene treatment subgroup.Conclusion:rIL-10 gene treatment attenuates the area of collagen deposition in liver fibrosis rats associated with downregulation of TIMP 1.

Objective To construct and identify recombinant expression plasmid of small interfering RNA (siRNA)targeting hepatitis B virus X protein(HBx), and observe its effect on mitoehondrial function in healthy liver cell line steadily expressed HBx gene (HL-7702/HBx). Methods Two siRNA sequences containing short hairpin structure, which target on the total length HBx gene, were synthesized and cloned into the vector psiRNA-Hh1GFPzeo to eonstruct recombinant expression plasmids pX1 and pX2. Non-specific recombinant pScr plasmid served as control. After siRNA transfected into HL-7702/HBx cells line by liposome, reverse transcription-polymerase chain reaction (RT-PCR) and Western blot were performed to identify the suppressive effect on HBx expression. Levels of intraeellular reactive oxygen species (ROS) and mitochondrial membrane potential (△ m) were determined by flow cytometry. The experimental results were compared by analysis of variance. Results Successful constructions of pX1 and pX2 were confirmed by restriction enzyme digestion and sequencing. The expressions of HBx mRNA and protein after 48 h of transfection into HL-7702/HBx cells in control group were 0.65± 0.12 and 0.62± 0.09, respectively, which were both higher than those (0.33±0.10 and 0.19±0.08, respectively) in group pX1 (t=4.73, P<0.05; t=7.53, P<0.05) and those (0.48±0.10 and 0.37±0.11, respectively) in group pX2 (t=2.39, P<0.05;t=4.43,P<0.05). But the inhibition of group pX1 was stronger than that of pX2 (t=2.28,P<0.05). Levels of ROS and △ m after RNA interference were 5.00±0.38 and 33.86±0.50, respectively, while those in control group were 72. 10±0. 55 and 3. 57±0.26, respectively (ROS: t=276.22, P<0.05; △ m: t=107.15, P<0.05). Conclusions siRNA targeting HBx can efficiently and specifically suppress the HBx expression in HL-7702/HBx cells, and decrease the level of ROS and increase the level of △ m, thus relieve cellular oxidative stress.