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.

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.

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

Acquired Immunodeficiency Syndrome ; HIV Infections ; Humans

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

Objective To retrospectively summarize the normal reference range of trachea wall thickness,lumen diameter,wall area and wall area ratio[WA%=mean wall area/(mean wall area+lumen area)]of Chinese healthy adults,and its related factors. Also,to observe the difference of inner diameter between superior and inferior bronchus.Methods Based on computer measurement techniques of bronchus,a CT quantitative analysis was carried out in 701 cases of normal healthy people who had negative results in lung cancer screening of health examination at our hospital.Results The value of trachea wall thickness,lumen diameter,wall area and wall area ratio was(1.322 mm,18.024 mm,78.93 mm2 ,0.27)respectively.In different gender,the trachea wall thickness,lumen diameter,wall area and wall area ratio had statistical significance (P<0.05).Also,they had good consistency with gender (r=-0.512,-0.472,-0.559,0.315).In different gender and age,the difference of inner diameter between the superior bronchus and inferior bronchus was always a positive value.Conclusion The CT quantitative analysis method has advantages of convenience,direct-vie-wing and accuracy.It is good for quantitative detection and research of bronchus structure.Bronchial wall thickness,lumen diameter, wall area and wall area ratio have significant difference because of gender.The inner diameter of superior bronchus is always greater than that of the inferior bronchus.

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.

Objective To prospectively evaluate the mid-term results of mitral annuloplasty using autologous pericardium for mitral valve insufficiency.Methods From April 2004 to December 2011,48 patients underwent mitral annuloplasty using autologous pericardium,the average length of pericardial strips was (51.9 ±2.8)cm.Carpentier classification was class Ⅰ in 5 cases,class Ⅱ in 41 cases,and class Ⅲ in 2 cases.The mitral valve repair techniques included quadrangular resection in 30 cases,valve repair in 7 cases,chordae transposition in 4 cases,edge to edge technique in 2cases,and artificial chordae tendineae in 3 cases.Concomitant procedures included one case arotic valve replacement,11 cases tricuspid valve repair,9 cases coronary artery bypass grafting,1 case coronary artery muscle bridge resection,and 1 case permanent pacemaker implant.Datum on long-term outcomes were obtained by questionnaires and by phone interview [average follow-up time (62.2 ± 21.3) months].Results Compared with preoperative datum,the diameters of left ventricular end diastolic diameter (LVDD) and left atrial diameter (LAD) examined by echocardiography show significant reduction in postoperative [(58.6 ± 1.7) mm vs (45.1 ± 1.3) ram,t =12.85,P ＜0.01 ; (50.6 ± 1.6)mm vs (38.0 ± 1.4)ram,t =9.58,P ＜0.01].There was early postoperative death in one case,cerebral infarction in one case,but none of patients died in late postoperative period.One patient had moderate mitral valve regurgitation in long-term follow-up.None of patients had redo operation and hemolytic complications.Conclusions Mitral annuloplasty using an autologous pericardium was an acceptable technique with low anticoagulation complications,permanent,well left ventricular function maintenance,and an economic method.

Adult ; Humans ; Male ; Military Personnel ; Models, Biological ; Muscle Contraction ; physiology ; Physical Endurance ; physiology ; Reference Values

Animals ; Apoptosis ; physiology ; Brain Ischemia ; metabolism ; physiopathology ; Down-Regulation ; drug effects ; Ischemic Postconditioning ; methods ; Male ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; metabolism ; prevention & control ; p38 Mitogen-Activated Protein Kinases ; metabolism