1.The cloning,expression and purification of cDNA in E.coli
Xiaodong WU ; Lijuan XUE ; Shaojuan YANG ; Hui ZHU ; Weizhong WANG
Chinese Journal of Immunology 1986;0(04):-
Objective:The human heparin-binding growth factor Midkine(hMK) gene was constructed and expressed in E.coli, and the activity of MK protein was tested.Methods:The MK gene was amplificated from the tissue of human embryo kidney by RT-PCR techniques. The MK fragment without signal peptide was inserted into plasmid pET30a. The gene was expressed in E.coli, the protein was purificated by affinity chromatography, and the activity was tested by ~3H-TdR method.Results:The sequence of hMK in our experiment was as same as the sequence published in Genebank. The activity assay was shown high activity of MK protein in our experiment.Conclusion:The expression carrier of hMK was established successful and it can expression high activity hMK protein in E.coli.
2.Docetaxel combined with cisplatin and fluorouracil in the treatment of advanced gastric cancer
Hui LI ; Wenhui SUN ; Bei XUE ; Changping WU
China Oncology 2006;0(10):-
Background and purpose:Gastric cancer is one of the leading causes of cancer death throughout the world.When compared with optimal supportive care alone,combination chemotherapy yields a significant advantage in the management of advanced gastric cancer.However,no single regimen has emerged or been accepted as being clearly superior over another.Here we investigated the efficacy and safety of "docetaxel+ cisplatin + fluorouracil" 5-day combination chemotherapy as treatment in patients with advanced gastric cancer.Methods:Between 2004 January and 2005 July,we enrolled 30 patients [males 22,median age 53 years(range 28-72)] with advanced gastric cancer.The regimen consisted of docetaxel 75 mg/m~(2) on day 1,cisplatin 15 mg/ m~(2) on days 1-5,and fluorouracil 500 mg/ m~(2) on days 1-5,every 3 weeks.All patients received over two cycles of this regimen.Results:A total of 99 cycles were administered.Mean cycle number per patient was 3.3.The administered dose intensity of docetaxel was 23.5 mg/m~(2)/week,fluorouracil 735 mg/ m~(2)/week and cisplatin 14.7 mg/ m~(2)/week,which corresponded to 94%,97.5% and 92.3% of planned doses.Of these patients,16(53.3%) achieved a partial response,8(26.6%) stable disease,and 6 patients(20%) showed progressive disease.The median time to progression was 5.1 months.(95% CI 4.1-6.0 months).Median overall survival was 9.9 months.(95% CI 8.1-11.6 months).Leukopenia occurred during 36.7% of cycles(36 of 99 cycles);18.2% grade Ⅰ,10.1% grade Ⅱ,5.0% grade Ⅲ and 3.0% grade Ⅳ.Anemia occurred in 12.1%(12 of 99 cycles);7.1% grade Ⅰ and 5.0% grade Ⅱ.Thrombocytopenia was 15.2%(15 of 99 cycles) and all were grade Ⅰ or Ⅱ.Diarrhea,stomatitis and hypersensitivity occurred in 16.7%(5 out of 30 patients),respectively.Neutropenic fever occurred in two patients(6.7%) and myalgia in nine(30%).Conclusions:Docetaxel combined with fluorouracil and cisplatin is an active and tolerable regimen for the treatment of patients with advanced gastric cancer.
3.AdH1-siRNA-Survivin inhibits tumor growth and metastasis of mouse melanoma
Zhiyong YU ; Weigan SHEN ; Qingyu XUE ; Hui WU
Journal of Third Military Medical University 2003;0(18):-
Objective To evaluate the effect of adenovirus-mediated siRNA targeting Survivin on tumor growth and metastasis of mouse melanoma.Methods A replication-defective recombinant adenovirus AdH1-siRNA-Survivin targeting mouse Survivin was constructed using the homologous recombination technique.The expression of Survivin protein in B16BL6 cells after infection was detected by Western blotting.Mouse melanoma models were established by subcutaneous injection of B16BL6 cells.AdH1-siRNA-Survivin of 2?109 pfu every week by multi-spot intratumoral or peritumoral injection was performed to every mouse(totally three times).The tumor volume was measured every five days after tumor cell implantation,and the microvessel density(MVD) of tumor was counted by immunohistochemistry under the microscope.For the experimental metastasis assay,B16BL6 cells infected with AdH1-siRNA-Survivin were injected into the tail veins of mice,then the visible black nodules on the lung surface were counted macroscopically at 21th day after tumor cell injection.Results AdH1-siRNA-Survivin could obviously decrease the expression of Survivin protein in B16BL6 cells.When compared with the blank control and AdH1-empty control,the tumor growth was inhibited and MVD was less in the mice injected with AdH1-siRNA-Survivin(P
5.Surgical treatment of primary pulmonary artery sarcoma.
Chinese Medical Journal 2011;124(3):461-463
BACKGROUNDPulmonary artery sarcoma originating from the pulmonary artery is a rare disease and its prognosis is extremely poor. Most patients are initially thought to have pulmonary embolism.
METHODSFive patients (3 males and 2 females, mean age was (42.0 ± 8.5) years) of primary pulmonary artery sarcomas underwent surgical procedures. Sarcoma resection and right ventricle outflow tract reconstruction were performed.
RESULTSThere was no death 30 days post-operation except one patient. The function of heart was improved to NYHA II.
CONCLUSIONSGiven the significant morbidity and mortality of pulmonary artery sarcomas, an aggressive strategy for resection is indicated. Early diagnosis and complete surgical resection is perhaps the best way to improve patients' survival with pulmonary artery sarcoma.
Adult ; Female ; Humans ; Male ; Middle Aged ; Pulmonary Artery ; pathology ; surgery ; Sarcoma ; surgery ; Treatment Outcome
8.Study of nitrogen silicon zirconium-hydroxylapatite as a coating on pure titanium implantation.
Lu YIN ; Ying-xue HUI ; Jiang-wu YAO
West China Journal of Stomatology 2009;27(5):492-495
OBJECTIVETo compare the osseointegration and corrosion of different kind of coatings on pure titanium.
METHODSBy plasma spraying technology, nitrogen silicon zirconium-hydroxylapatite (ZrSiN-HA) compound coating was prepared on the surface of pure titanium and the pure titanium was implanted with the mandible of the experimental animal. The superficial appearance of the compound coating was observed and analyzed by scanning electron microscope (SEM). The ZrSiN-HA, nitrogen silicon zirconium (ZrSiN), hydroxylapatite (HA) and pure titanium were compared and examined. In the four groups, the bone binding force of coatings were detected, and the superficial appearance of the break regions were observed by the electronic multipurpose material testing aircraft. In the four groups, the anticorrosive of coatings were compared and examined on the electrochemistry corrosion testing system.
RESULTSExperimental results indicated that the implant surface sprayed with the ZrSiN-HA was more compact than that sprayed with HA. The crystallization was obvious. Compared with the other coatings, the bone binding force of the ZrSiN-HA coating was the highest, and its anticorrosive performance was the strongest.
CONCLUSIONThe application of ZrSiN-HA coating is advantageous to the long-term retention of implant, and it has huge latent application value to dental dummy.
Animals ; Coated Materials, Biocompatible ; Durapatite ; Materials Testing ; Microscopy, Electron, Scanning ; Nitrogen ; Osseointegration ; Prostheses and Implants ; Silicon ; Surface Properties ; Titanium ; Zirconium
10.The effects of Stattic, a STAT3 inhibitor, on the growth, migration and radiosensitivity of liver cancer cells Bel-7402
Lihua ZHU ; Gang XU ; Aihua GONG ; Yan WANG ; Hui XUE ; Li ZHANG ; Chaoyang WU
Chinese Journal of Radiological Medicine and Protection 2015;35(6):413-418
Objective To study the effects and preliminary mechanism of Stattic (Y705),an inhibitor of the signal transducer and activator of transcription-3 (STAT3),on the growth,migration,invasion and radiosensitization of hepatocellular carcinoma cell line Bel-7402.Methods Bel-7402 cells were divided to four groups:blank control group,Stattic treatment group,radiation group,and Stattic combined with radiation group.The cell growth and proliferation were detected by using CCK8 kit.The influence of Stattic on radiation sensitivity of Bel-7402 cells was determined by clone formation assay.The cell migration and invasion ability were tested by scratch migration assay and transwell assay,respectively.The protein expressions of STAT3,p-STAT3,Bax,Bcl-2,Caspase-3,Cleaved Caspase-3,MMP-2 and MMP-9 were quantified by Western blotting assay.Results Stattic significantly inhibited the growth of human hepatocellular carcinoma Bel-7402 cells with a dose-depended manner.The IC50 of Stattic after 48 h treatment was 2.5 μ mol/L.When 1.0 μmol/L Stattic was combined with 8 Gy X-rays,there was a synergistic effect in inhibition of cell proliferation with a inhibition rate of (15.00 ± 1.87) % (F =63.30,P < 0.05).Scratch migration assay and transwell invasion assay showed that the migration and invasion abilities of the combination group were significantly reduced.In addition,compared with the radiation group,the SF2,D0and Dq values obtained from survival curve were decreased (t =4.20,6.92,9.32,P <0.05),the protein expressions of p-STAT3,MMP-2,MMP-9 were reduced (t =5.32,6.02,13.26,P <0.05),the protein expressions of Bax and Cleaved Caspase-3 were increased in the combination group(t =-7.82,-14.09,P < 0.05),meanwhile the protein expressions of Bcl-2 was decreased (t =18.43,P < 0.05).When the concentration of Stattic was 0.5 μmol/L,the radiation sensitization ratio at 2 Gy (SERSF2) was 1.22.Conclusions By inhibiting the activation of the p-STAT3 in Bel-7402 cells,stattic could induce cell apoptosis and increase the radiosensitivity,down regulate MMP-2 and MMP-9 and thereby reduce the invasion and migration of tumor cells.