Objective To investigate the role of PI3K/AKT pathway on growth and invasion effect in human glioma cells U251 by Ad-PTEN and phosphatidylino-sitol 3-kinase inhibitor LY294002.Methods Western blotting analysis was used to detect the PTEN、PI3K and AKT protein expression after LY294002 and Ad-PTEN were transfected into glioma cell lines U251.Flow cytometry,Annexin V/FITC methods,MTT methods were used to detect the proliferative activitity of the U251 cells.The invasion of U251 cells in vitro was examined by Scrath assay and Transwell cell invasion test.The difference between the combined therapy group and the single LY294002 therapy group was also examined.The downstream protein expression in PI3K/AKT signal pathway were also detected by Western blotting.Results The protein expression of PTEN in combined group were significantly higher,while PI3K、AKT(p-ser473)was significantly lower than that in the other three groups(P＜0.05).After treating,the cells in combined group were inhibited at G₀/G₁ phase,and the cell number decreased at S phase.The apoptosis rate in combined group was higher than that in DMSO、Ad-nonsense and LY294002 group.As compared with DMSO、Ad-nonsense and LY294002 group,the cell multiplication rate in combined group presented decreasing trend since the second day in culture(P＜0.05).The invasion and migration cells in combined group was lower than that in other three groups.The protein expression of PCNA、CyclinD1、Bcl-2、NF-κB、MMP-2、MMP-9、and FAK was significantly lower than that in the other three groups(P＜0.05).Conclusion The combined therapy of Ad-PTEN and LY294002 plays a synergistic role in inhibiting the proliferation and invasion of U251 cells.

Objective To study the expressing of recombinant adenovirus with exogenesis genes in AGZY and the genetic stability.Method Transfected AGZY with two kinds of rAd.Then,detect the mRNA expression of Gene hIL-2 and Gene hGM-CSF with half-quantity RT-PCR.Result The expression level of hIL-2 and hGM-CSF in AGZY increased with time upto 72h.Conclusion The AGZY passage has no affect on the expression efficiency and the genetic stability of rAd-IL、rAd-GM-CS.

Objective To explore effects of norcantharidin(NCTD)on the proliferation and killing activity of cytokine-induced killer cells(CIK).Methods Cord blood mononuclear cells were extracted by Ficoll density gradient method and then induced into CIK cells.The experiment was divided into NCTD-CIK group with a final concentration of 30μmol / L NCTD and control group CIK without NCTD.Changes in the proliferation of CIK cells,the immune phenotype and cytotoxic activity were observed using cell counting,flow cytometry and MTT method.Results The rate of cell proliferation and anti-tumor activity in NCTD-CIK group were significantly higher than those of the control CIK group,which showed statistical significance(P＜0.05).Conclusion NCTD promoted the expansion of CIK cells and enhanced their anti-tumor activity,which will provide a new approach for adoptive immunotherapy of CIK.

Objective To evaluate the value of differential diagnosis in solitary pulmonary nodule(SPN)with HRCT and ¹⁸F-FDG PET/CT.Methods 60 solitary pulmonary nodules were examined with ¹⁸F-FDG PET/CT and local thin-slice high resolution CT(LTHRCT,for short HRCT),all of which were proven pathologically.The diagnosis of PET/CT relied on the metabolism feature of PET and the morphology feature in low-dose CT during quiet respiration.The diagnosis of PET/CT combining HRCT considered the morphology feature of HRCT on the basis of PET/CT.Results The sensitivity,specificity,accuracy of PET/CT were 89.5%,77.3% and 85.0% respectively.accordingly,the sensitivity,specificity,accuracy of PET/CT combining HRCT were 97.4%,81.8% and 91.7% respectively.Conclusions PET/CT combining HRCT can raise the sensitivity,specificity and accuracy of PET/CT in the diagnosis of SPN.

Objective To explore the difference of serum protein mass spectrometry between the sensitive and drug resistance lung adenocarcinoma patients,and to discover serum protein markers for predicting chemotherapy sensitivity of lung adenocarcinoma patients.Methods To detect the protein mass spectrometry of 22 chemosensitive lung adenocarcinoma patients and 21 chemotherapy-resistant cases with WCX2 protein chip combining with SELDI-TOF-MS technique,and then screen differentially expressed proteins.Results In this experiment,7 differentially expressed proteins were screened,which can differentiate the chemosensitive or the chemotherapy-resistant patients with standard platinum-based chemotherapy.The m/z value of them is 16083.50Da,7963.17Da,16121.50Da,15921.60Da,8066.79Da,8040.17Da and 3394.57Da respectively.Predictive model with M/Z 7963.17Da and 16083.50Da could differentiate the chemosensitive or the chemotherapy-resistant patients,whose sensitivity was 86.36% and specificity was 80.95%.Conclusions SELDI-TOF-MS is an effective instrument to find sensitive serum marker for predicting lung adenocarcinoma chemotherapy sensitivity.

Objective To study the expression of Cry61(Cysteine-rich 61)in epithelial ovarian cancer and its relationship with angiogenesis.Methods Using semi-quantitative reverse transcription-polymerase chain reaction(RT-PCR)and immunohistochemistry,the mRNA and protein expression of Cyr61 were examined in 40 ovarian epithelial cancers and 10 normal ovarian tissues,and using immunohistochemistry examine MVD in the tissues above.Results The expression of Cyr61 and MVD in tissues of ovarian epithelial cancer were higher than those in normal ovarian tissues,and had significant correlation with phases and metastasis of carcinoma.Increase of MVD was in accordance with the up-expression of Cyr61 in tumor tissue.Conclusions Cyr61 plays an important role in the pathogenesis and development of ovarian epithelial cancer,Cyr61 may play a role in ovarian carcinogenesis by stimulating angiogenesis.

Objective To investigate the expression of MMP-2 and TIMP-2 mRNA in transitional cell carcinoma of bladder and discuss their clinical significance.Methods Using RT-PCR and real time quantitative PCR(RQ-PCR)technique,the expression of MMP-2 and TIMP-2 mRNA of 45 cases of bladder carcinoma(tumor group)and 10 cases of normal bladder tissue(control group)were analyzed.Results MMP-2 and TIMP-2 were not expressed in control group.MMP-2 was expressed in 30 cases tumor samples and TIMP-2 was expressed in 26 cases.The expression of MMP-2 mRNA in superficial bladder cancer and G1 cancer were lower than that in infiltrating bladder cancer and G2-G3 cancer respectively.The differences were statistically significant(P ＜0.05).The expression of MMP-2 in recurrence group was higher than that in incipient group.TIMP-2 mRNA expression decreased with grades and stage.The expression of TIMP-2 in superficial bladder cancer and G1 cancer were higher than that in infiltrating bladder cancer and G2-G3 cancer respectively.There was statistical difference between two groups(P ＜0.05).TIMP-2 expression in incipient group was higher than that in recurrence group,but the difference was not significant(P＞0.05).Conclusion These results suggest that MMP-2 and TIMP-2 play an important role in the invasive step of transitional cell carcinoma of bladder.MMP-2 may become a new approach to the diagnosis of bladder carcinomas.

Objective To investigate the expression of Ezrin in colorectal carcinoma and the relationship between Ezrin and epithelial mesenchymal transition of colorectal carcinoma.Methods By SP immunohistochemical method,we determined Ezrin,E-cad and Vimentin expressions and the relationship between them in 64 specimens of coloretal carcinoma and 20 specimens of nomal colorectal tissues.Results The positive rate of Ezrin in colorectal carcinoma was significantly higher than that in the normal tissue(73.4% vs 15%;P＜0.01).The positive rate of E-cad in cancerous tissue was lower than that in the normal tissue(48.4% vs 80%;P=0.028).The positive rate of Vimentin in colorectal carcinoma was significantly higher than that in the normal tissue(20.3% vs 0.00%;Fisher′s exact test,P=0.032).The overexpression of Ezrin was correlated with the clinical stage and age(P＜0.05).The Spearman correlation analysis demonstrated that Ezrin expression was negatively correlated with E-cad expression,while it was not correlated with Vimentin expression.Conclusions Ezrin may be not associated with epithelial mesenchymal transition of colorectal carcinoma,but it may play a crucial role in the progression of colorectal carcinoma.

Objective The aim of this study was to evaluate the efficacy and safety of combination therapy with docetaxel and nedaplatin in advanced esophageal cancer.Methods 45 patients were included in the study(30 male,15 female);median age:54 years(range 37-72);37 cases of squamous cell carcinoma,8 cases of adenocarcinoma.10 patients never received treatment,35 re-treatment patients who received more than one chemotherapy.Patients received docetaxel 37.5mg/ m² on day 1、8,nedaplatin 80mg/ m² on day 2,every 3 weeks.Evaluate the efficacy and toxicity every 2 cycles according to WHO criteria.Results A total of 177 chemotherapy cycles was administered.The median number of cycles was 3.9(range 1-6).The response rate was 37.8% including one complete response,22 partial response.Initial treatment or retreatment,stage,histological type had no effect on short-term effect.Common toxicity was neutropenia.Conclusion the combination chemotherapy of docetaxel and nedaplatin is an active regimen with moderate toxicity in the treatment of patients with progression esophageal cancer.

Objective To evalue recent efficiency and side effects of ¹²⁵I seed brachytherapy for late-staged malignant tumors.Methods All the patients with residual or recurrent unresectable malignancies were treated with ¹²⁵I seed implantation under CT scan after they were given a local anesthesia.The seeds were retrogradely placed with a gun applicator according to the previous therapy planning.Results All the patients were implanted succeedly and no severe complications were recorded postoperatively.In all cases,the particles didn′t fall off or shift.Only minority of patients showed light loss of thrombocyte and hemoglobin.Acute leucocytopenia was seen in 3 patients.A complete response was achieved in 5 patients and a partial response in 11 patients.6 patients were assessed as having stable disease while 1 patients having progressive disease.In three patients the serum TSGF level was increased significantly,which was supposed to be failure.The local control rate was 82.6%(19/23)and the total response rate was 69.6%(16/23).Thirty patients suffered the pain before treated with¹²⁵I.Postoperatively a complete pain relief was achieved in 17 patients,a partial relief in 9 patients and no change in 4 patients.The response rate was 86.7%(26/30).The survival rate of 1,3,6 month and 1 year were 100%,94.59%,81.08% and 62.16% respectively.Conclusions ¹²⁵I brachytherapy for late-staged malignant tumors gives a wide range,definite recent efficiency and satisfactory reliability.However,own to all the patients are very late-staged,a high survival rate have not been obtained.

Objective To study the clinical effect of bronchial artery infusion(BAI)chemotherapy combined with systemic chemotherapy for patient with advanced non-small cell lung cancer.Methods 138 cases with advanced non-small cell lung cancer were divided into two groups.70 cases(A group)received BAI chemotherapy conbimed with systemic chemotherapy.68 cases(B group)were treated with systemic chemotherapy alone.The same systemic chemotherapy was taken in the two groups.The clinical data were analyzed retrospectively.Results Short-term response rate(61.42%),time to progression(TTP)(125.23 days),half-year survival rate(94.29%)and 1-year survival rate(94.29%)in BAI chemotherapy group were significantly higher than those in control group(46.27%,85.94 days,82.35%,42.65% respectively).Conclusion BAI chemotherapy combined with systemic chemotherapy is helpful for patient with advanced non-small cell lung cancer to improve therapeutic effect,to prolong survival time and improve quality of life.

Objective To evaluate the outcome of locally advanced non-small cell lung cancer treated with radiotherapy and chemotherapy.Methods From July 2001 to December 2006,83 patients with locally advanced non-small cell lung cancer were enrolled.Three groups were divided based on time and sequence of radiotherapy and chemotherapy.All patients used three dimensional conformal radiation therapy and cisplatin based chemotherapy.The short-term response and 1-year,2-year survival rates,median survival time and toxicity in three groups were compared.Results The overall response in three groups were 89.3%,78.1% and 68.5%(P＞0.05)respectively.The median survival time were 21 months,18 months and 16 months,respectively,The 1-,2-year survival rate were 75%,67.86%,51.16% and 49.01%,44.44%,31.97%(P＜0.05)respectively for three groups.And the differences of toxicity rates for three groups were not statistically significant.Conclusion Induction chemotherapy followed by three dimensional conformal radiation therapy was efficacy and safety.The sequential chemoradiotherapy improved the overall survival of locally advanced non-small cell lung cancer.The output indicated early irradiation with chemoradiotherapy is superior to late irradiation.Induction chemotherapy is optimization for one or two cycles.

Objective To study the loads of HR-HPV in cervical diseases,and to explore the relationship between high viral loads of HR-HPV infection and cervical carcinoma,cervical intraepithelial neoplasia(CIN).Methods The study was carried out in Liaoning Cancer Hospital,3676 women who received treatment were recruited HPV DNA by hybrid capture Ⅱ(HC-Ⅱ),then they were divided into 4 groups basing on the different levels of the viral loads of HPV DNA:negative(RLU/CO＜1.0),low viral loads(1.0≤RLU/CO＜10),medium viral loads(10≤RLU/CO＜100),high viral loads(RLU/CO≥100).The relationship between viral loads and various cervical diseases were analyzed by unconditional multi-normial logistic regression.Results In CIN1 group,there was 23.07% of patients in high viral loads,the odds ratio was only 9.42(95%CI:6.84-12.99).In CIN2 group,the proportion of high viral loads accounted for 30.84% with odds ratio to 22.55(95%CI14.88-34.18:).In CIN3 group,the proportion of high viral loads accounted for 47.89% with odds ratio up to 82.13(95%CI:52.69-128.03).In cervical carcinoma group,the proportion of high viral loads was 51.66% with odds ratio more higher up to 161.90(95%CI:109.45-239.50).Conclusion There is significant correlation between the viral loads of HR-HPV and severity of cervical lesion.High viral loads of HR-HPV which persistently infects predicts high degree of cervical diseases and cervical carcinoma.

Objective To compare the therapeutic effect of Ultrasonically Activated Scalpel(UHS)with electrotome in axillary lymph node dissection(ALND)of breast cancer.Methods A retrospective analysis was performed on 105 patients who received modified radical mastectomy.And 41 cases only subject to UHS,64 cases only subject to electrotome.We observed the difference of operation time,blood loss of ALND,postoperative draining volume,duration of axillary draining and length of stay and postoperative complications.Results There was no significant difference in operating time,blood loss and length of stay between the two groups.However,the incidence of postoperative complications such as flap necrosis and postoperative rehaemorrhagia in UHS group was dramatically decreased as compared with electrotome group.Conclusion The therapeutic effect of UHS is better than that of elect rotome in modified radical mastectomy.

Objective To examine the expression level of let-7 precursor and K-ras in lung adenocarcinoma patients by real time-PCR,in order to explore their roles in lung adenocarcinoma initiation and development.Methods GAPDH was adopted as a control,we use real time PCR to examine the expression of let-7 precursor and K-ras in carcinomatous tissue and the around normal tissue respectively in 31 selected lung adenocarcinoma patients.Results Compared to the normal tissue,let-7 precursor expression was reduced about 61.29%,while K-ras expression were increased about 67.74%.The measured value of let-7 precursor and K-ras in carcinomatous tissue and the around normal tissue respectively carry on statistics analysis.Their difference has the remarkable statistics significance.The correlation between let-7 precursor and K-ras in the expression of carcinomatous tissue has the remarkable statistics significance.Conclusion Those results suggest that let-7 precursor or/and K-ras may play an certain cooperated role in the initiation and development of lung adenocarcinoma.

Objective To evaluate the objective response rate,time to progression(TTP)and toxicity between Docetaxel/Capecitabine and Oxaliplatin/Capecitabine in the treatment of advanced gastric cancer.Methods 21 patients with advanced gastric cancer were administrated with Docetaxel 75mg/m²(d1),Capecitabine 1700mg/m²/d divided in two daily doses given from d1 to d14.The cycles were repeated every 21 days.All patients received two to six cycles.The other 20 patients with advanced gastric cancer were administrated with Oxaliplatin 130mg/m²,(d1)combined Capecitabine 1700mg/m²/d divided in two daily doses given from d1 to d14.All patients received two to six cycles.The treatment was finished after 2 recycles.Results The total response rate in Docetaxel/Capecitabine was 52.4 %,the most common adverse effects were febrile neutropeni,neutropenia,alopecia,diarrhea and inflammation of oral mucosa.The response rate in XELOX was 50.0 %,the most common adverse effects were hand foot syndrome and peripheral neuropathy.Conclusion The Oxaliplatin/Capecitabine regimen offers a better therapeutic option in patients with advanced gastric cancer.