Objective Construct a recombinant adenovirus containing the human hydroperoxidase(hTPO)gene,transfer the hTPO genes and human sodium/iodide symporter(hNIS)gene into glioma cells and study the iodide uptake ability of the cells and the killing effect of radioiodine,so as to investigate the feasibility of radioiodine therapy on glioma.Methods The recombinant adenovirus AdTPO was constructed by AdEasy system.Tranfecting hNIS gene into human glioma cell line U251 through recombinant plasmid,the stably expressing hNIS gene cell line hNIS-U251 was determined as negative control group.Then hTPO gene was transfected into hNIS-U251 by recombinant adenovirus AdTPO,achieving AdTPO-hNIS-U251,determined as testing group.U251 cell line without hNIS or hTPO gene was applied as control group.We investigated the iodide uptake assay,perchlorate suppressive assay,iodide organification assay to confirm gene expression and function.Killing effect of radioiodide on tumor cells was studied by clonogenic assay in vitro.Results Recombinant adenovirus AdTPO was correctly constructed.we were successful in co-transfecting hTPO gene into stably expressing hNIS cell line(hNIS-U251),obtaining AdTPO-hNIS-U251.In hNIS-U251 group,the iodide uptake ability(55769.96±4353.26count·min^-1)was 110 folds higher than that in control group(507.67±57.69count·min^-1),the effective half-life time was 7 minutes,the organification percentage was 0.1%,and the rate of clonal forming was(9.08±2.86)%,that was 10 folds lower compared with control group.While in AdTPO-hNIS-U251 group,the iodide uptake ability(74647.53±3605.88count·min^-1)was 147 folds higher than that in control group,the effective half-life time prolonged to 13 minutes,the organification percentage reached to 10%,and the rate of clonal forming was(6.80±2.09)%,which was 13 folds lower than control group respectively.Conclusion By co-transfection of hTPO and hNIS genes into glioma cell line U251,the iodide uptake ability increased markedly,hTPO increased the retension of iodide in the cells,and the ¹³¹I could kill tumor cells effectively.

Objective To investigate the mRNA expression of tumor suppressor gene ING1 in breast carcinoma and evaluate its clinical significance.Methods Reverse transcription polymerase chain reaction(RT-PCR)was used to detect the mRNA expression of ING1 in 51 cases of breast carcinoma,30 adjacent normal tissues and 12 benign breast tumors.The relationship of ING1 mRNA expression to clinicopathological characteristics was statistically analyzed.Results (1)The expression of ING1 mRNA in breast carcinoma tissues(0.5458±0.2310)was lower than that in the adjacent normal tissues(0.6625±0.1699).The expression of ING1 mRNA in groups of breast carcinoma tissues,the adjacent normal tissues and the benign breast tumor tissues were significantly different(P＜0.05).(2)ING1 mRNA expression in breast carcinoma tissues was related with pathological type and lymph node metastasis(P＜0.05).ING1 mRNA expression in breast carcinoma tissues had no relevance with the other clinicopathological characteristics(P＞0.05).Conclusion The mRNA expression of ING1 is significant associated with tumor progression in breast carcinoma,and it may be considered as a valuable marker for the gene therapy of breast carcinoma

Objective To investigate the effect of Pine Cone Extract(PCE)on the mice with transplanted S180 sarcoma.Methods On the second day after S180 sarcomas being transplanted into the right axilla of mice,the mice were divided into four groups(water control group,cyclophosphamide group,low PCE group and high PCE group)and administered water,cyclophosphamide(ip),low PCE group and high PCE by gavage for 14 days,respectively.The dose volume was 0.02mL/g body weight and once daily.Another group was used as normal control.Then indices of thymus and spleen and inhibitory rate of S180 tumor were observed.Results The indices of the thymus and spleen in the tumor PCE group had no significant difference compared with water control group.Inhibition rate of S180 sarcoma in the high PCE group compared with the control group was 46.1%.Conclusion PCE obviously inhibits the growth of S180 sarcoma with 14 days administration at suitable dosage.

Objective To observe the function of calcium sensing receptor(CaSR)in differentiated THP-1(human acute monocytic leukemia cell line)cells.Methods In the present study,we induced and differentiated the monocyte-like THP-1 cells into macrophage-like THP-1 cells with PMA.The secretion of cytokines was analyzed by ELISA.The expression of CaSR and NF-κB in the differentiated THP-1 cells was identified by Western blotting.Results (1)We found that GdCl₃ elevation stimulated both of IL-1β and TNFα release in differentiated THP-1 cells.(2)GdCl₃ could increase the expression of NF-κB-p65 and decrease the expression of NF-κB-IκBα.The effects of GdCl₃ were inhibited by NPS2390.Conclusion The activation of CaSR in macrophage-like THP-1 cells could commit to cytokine secretion via NF-κB signal transduction pathway.These results suggested that CaSR might be involved in a variety of pathological processes mediated by activated monocyte-macrophages.

Objective To study the expression of Pokemon、Ki-67 and its clinical significance in gastric cancer and normal gastric mucosa.Methods The Pokemon and Ki-67 in gastric cancer(n=125),normal gastric mucosa(n=25)was detected by RT-PCR.Results Pokemon protein expression was high in gastric cancer group,which was significantly different from the expression in normal gastric mucosa.It was highly expressed in gastric cancer and was related with stage of TNM and lymph node metastasis,but were not related with the age,sex and the size of tumor.Conclusion Pokemon and Ki-67 protein expression may be correlate with the carcinogenesis and genesis,development and metastasis of gastric cancer.Pokemon may be an effective target gene for therapy of gastric cancer,accordingly,which may develop a new way of gene therapy for gastric cancer.

Purpose To explore the expression,relation and clinical significance of the apoptotic inhibitors Survivin and Livin mRNA in breast cancer.Method The mRNA expression of Surviving and Livin were detected in breast cancer samples and matched para-cancerous normal tissues by real-time quantitative RT-PCR(qRT-PCR).Results The positive rates for Survivin and Livin mRNA expressions were 62.3%(38)and 54.1%(33)in 61 breast cancer tissues,respectively.The significant correlation of Survivin was found with clinical stage,lymph node metastasis and HER-2 expression(P＜0.05).And there was a relationship between the expression of Livin and lymph node metastasis.There was no correlation between mRNA expression of Survivin and Livin.It was shown that patients with high Survivin and Livin mRNA expression had significantly poorer overall survival compared with patients with low expression of Livin and Survivin(P＜0.001 and P＜0.001,respectively).Conclusions Our study suggests that up-regulated mRNA expression of Survivin and Livin genes may closely correlated with the carcinogenesis and development of breast cancer.The relationship between Survivin or Livin and lymph node metastasis indicates that higher Survivin and Livin expressions may imply patient′s poor prognosis,and Survivin and Livin may serve as a new target for the therapy of breast cancer.

Objective To observe the expression differences of vascular endothelial growth factor(VEGF)and tumor suppressor gene P16 between the two tumor samples of hepatocellular carcinoma which has been separated respectively by transcatheter arterial chemoembolization(TACE)and surgical resection alone,and to study the effects of TACE on long-term prognosis of liver cancer.Methods The 65 paraffin embedded tissues blocks of hepatocellular carcinoma which have been pathologically confirmed were selected randomly.Among them,the 35 specimens were got by transcatheter arterial chemoembolization(TACE group),and the other 30 were separated by surgery without any treatment.The levels of VEGF and tumor suppressor gene P16 of the specimens were detected by immunohistochemical technique and comparative analysis.Results The expression of VEGF and P16 in TACE group was significantly higher than that of control group.Conclusion The high expression of VEGF of hepatocellular carcinoma after TACE may promote tumor recurrence and metastasis.However,the high expression of P16 may play a role in the inhibition of VEGF,thereby inhibiting tumor recurrence and metastasis,and there is no correlation between the expression of VEGF and P16.

Objective To investigate the expression of RASSF1A gene in human breast cancer and evaluate their clinical significance.Methods RT-PCR was used to detect the expression of RASSF1A mRNA in 51 cases of breast carcinoma,30 cases of normal tissue adjacent to breast cancers and 12 cases of breast benign diseases.Results (1)There is no difference between the expression of RASSF1A mRNA in normal tissue adjacent to breast cancers and breast benign diseases(P＞0.05).The deficient expression rate of RASSF1A mRNA in breast cancer is 51.0%(26/51)and lower than normal tissue adjacant to breast cancers and breast benign diseases(P＜0.01);(2)In breast cancer,loss of RASSF1A mRNA expression had significantly association with lymph node metastasis(P＜0.05);(3)Low or loss of RASSF1A mRNA expression in breast cancer had no association with age of patients,tumor size,tumor pathological type,the clinical TNM stage,ER,PR and HER-2 expressions(P＞0.05).Conclusion Loss of RASSF1A gene expression may play an important role in tumor progression,and may be considered as avaluable marker for predicting the prognosis in breast cancer.

Objective To study the expressions and clinical significance of VEGF,Ki-67 and P53 in human non-small cell lung cancer(NSCLC).Methods The expressions of VEGF,Ki-67 and P53 were detected by immunohistochemical method in 78 NSCLC,and the clinicalpathological features of lung cancer were analyzed statistically.Results Positive rates of VEGF,Ki-67 and P53 were 67%,71% and 50%,respectively.VEGF,Ki-67 and P53 expressions were close related with pathology type and TNM staging(P＜0.05).VEGF,Ki-67 and P53 expressions had no relationship with histological substype,lymph node metastasis status and patients′age(P＞0.05).The survival time of VEGF,Ki-67 and P53 positive expression groups was shorter than that of negative expression groups(P＜0.05).Conclusion VEGF,Ki-67 and P53 expressions may play an important role in the pathogenesis and progression of NSCLC.They have important prognostic value in NSCLC.

Objective To study the expression and significance of coagulation and fibrinolysis indexes in lung caner patients with different pathology types、stages and therapeutic states.Method The plasma levels of platelet(PLT),prothrobin time(PT),activated partial thromboplastin time(APTT),throbin time(TT),D-dimer(D-D),fibrinogen(FIB),tissue type plasminogen activator(tPA) and urokinase type plasminogen activator(uPA)and urokinase plasminogen activator receptor(uPAR)were measured by ELLISA technique in 62 lung cancer patients before and after therapy.Result The levels of PLT,PT,APTT,TT were no difference in all kinds of patients(P＞0.05).The levels of FIB were significantly higher in patients with Ⅳ stage and small cell lung cancer(SCLC)than those in other stages and types (P=0.028 and P=0.005,respedida)and 0.005 and the differences have statistical significance.The level of D-D was higher in adenocarcinoma than those in other types(P=0.005).The level of uPAR was lower in adenocarcinoma than those in other types and higher in Ⅲ stage patients than Ⅳ stage,bat the uPAR levels of patients who received operation and chemotherapy were lower than those not receiving any therapy (P=0.03,P=0.045 and P=0.007,respedively).The tPA level was higher in SCLC than those in other types(P=0.04).Conclusion The plasma level of UPAR was lower in patients who received any therapy than those not.The plasma of D-dimer is higher in adenocarcinoma patients than those in other type patients;Different type and clinical stage of lung cancer expressed different coagulation and fibrinolysis factors and quantity.