Objective:To prepare bifunction antibody of anti-human laryngocarcinoma/anti-CD 3 for the active immune therapy of laryngocarcinoma.Methods:To corss link monoclonal antibody of anti-human laryngocarcinoma and anti-CD 3 into bifunction antibody by chemical method,which can guide the killer function on carcinoma cells.Results:The killer rate of effector cells guided by bifunction antibody is higher than that of anti-CD 3 which can arise with the elevation of effect target ratio.Conclusion:Bifunction antibody of anti-human laryngocarcinoma/anti-CD 3 prepared by chemical cross linking method has the potential value for clinical application. [

Objective To investigate the effect of radiation on the mitochondrion in adenocarcinoma A549 cells.Methods After A549 cells were irradiated with 0,0.5,3 or 8 Gy of 60Co γrays,mitochondrion membrane potential of A549 cells was detected by JC-1 probe,and ATP activity was measured by ATP kit in a chemiluminescence apparatus.The mitochondria DNA copy numbers was detected by real-time PCR assay.Results At 24 h after radiation,the mitochondrion membrane potential of A549 cells in all the irradiated groups changed significantly (F =243.44,P ＜ 0.05),among which 0.5 or 3 Gy of radiation resulted in a significant increase of mitochondrion membrane potential of A549 cells (t =-10.12,-5.59,P ＜ 0.05).However,the mitochondrion membrane potential of A549 cells exposed to 8 Gy of radiation decreased significantly 24 h after radiation (t =15.22,P ＜ 0.05).The mitochondrion membrane potential of A549 cells in all radiation groups returned to the normal level 48 h after radiation (F =10.36,P ＜ 0.05).24 h after radiation,the level of ATP in A549 cells significantly changed respectively(F =97.08,P ＜ 0.05),similar to the mitochondrion membrane potential.The ATP level in 0.5 and 3 Gy groups increased significantly (t =1.66,7.27,P ＜ 0.05),and the level of ATP in 8 Gy group decreased significantly (t =-8.24,P ＜ 0.05).Furthermore,48 h after both 0.5 and 3 Gy of radiation,the ATP content in A549 cells was still higher than that in untreated A549 cells (t =4.60,8.53,P ＜0.05).The mitochondria DNA copy numbers in A549 cells increased significantly in all the radiation groups (F =118.00,P ＜ 0.05).Compared with untreated A549 cells the mitochondria DNA copy numbers in A549 cells increased at 0.5 Gy by 12 times(t =0.02,P ＜0.05),and increased at 3 and 8 Gy by 7 and 10 times,respectively (t =9.68,15.10,P ＜ 0.05).Conclusions High dose of radiation resulted in the decrease of mitochondrion membrane potential of A549 cells,which subsequently affected the production of ATP.However,radiation with moderate and lower dose could lead to the compensatory increase of mitochondrion membrane potential of A549 cells,which promoted the production of ATP.The mitochondria DNA copy numbers compensatory would increase after A549 cells were exposed to radiation within 8 Gy.

BACKGROUND: Adhesion molecules are closely associated with inflammation. Inflammation due to white blood cell (WBC) infiltration and free radical injury following brain ischemia are believed to be important factors contributing to the pathogenesis of multi-infarct dementia.OBJECTIVE: To determine the level of serum adhesion molecules and free radicals in patients with multi-infarct dementia to explore the relationship between their levels and multi-infarct dementia.DESIGN: A case-control trial.SETTING: Department of Neurology, Second Affiliated Hospital of Jilin University.PARTICIPANTS: Totally 82 patients with multi-infarct dementia were hospitalized in the Department of Neurology, Second Affiliated Hospital of Jilin University between January 2000 and December 2004. These patients included 32 cases of mild dementia, 21 of moderate dementia, and 29 of severer dementia. The normal controls were 23 concomitant healthy volunteers who came for routine physical examination.METHODS: Enzyme-linked immunosorbent assay (ELISA) and electron spin resonance were used to determine the level of serum soluble intercellular adhesion molecules (ICAM) and vascular cell adhesion molecules (VCAM), as well as oxygen free radical concentration in the normal controls and patients with multi-infarct dementia, and the association between the severity of the illness and the levels of adhesion molecules and oxygen free radicals was analyzed.MAIN OUTCOME MEASURES: Serum levels of ICAM-1 and VCAM-1 and oxygen free radical concentration in the two groups.RESULTS: Totally 82 patients with multi-infarct dementia and 23 healthy controls were included in this study and all enter the result analysis. In multi-infarct dementia patients, the serum levels of soluble ICAM-1 and VCAM-1 and oxygen free radical concentration [(469.00±76.33), (196.00± 45.91) and (1 103.30±98.96) μg/L, respectively] were significantly higher than those of the control group [(601.00±76.30), (4.018±1.656), and (1.295±0.718) μg/g, respectively, t=5.517-6.754, P ＜ 0.01], and the 3 indices were positively correlated with the severity of dementia (r=0.659 4,r=0.697 2, r=0.649 4, respectively, P ＜ 0.05); serum ICAM-1 and VCAM-1 levels were positively correlated with the concentration of oxygen free radicals (r=0.714 7, r=0.732 4, respectively, P ＜ 0.01).CONCLUSION: Serum ICAM-1, VCAM-1 and oxygen free radicals might be implicated in the pathophysiological development of multi-infarct dementia, and their levels increase in parallel with the severity of dementia.

Objective To investigate the proliferation inhibition of chlorpromazine combined with taxol on Hep-2 cells (human laryngeal carcinoma cell line) and the effects on cell cycle progression. Methods Hep-2 cells at logarithmic growth phase were divided into taxol groups(3.0,6.0 and 12.0 mg?L-1),chlorpromazine(12.0 mg?L-1) comined with taxol (4 mg?L-1)group and control group (100 mL culture fluid).MTT and flow cytometry were used to detect the proliferation inhibition rates of Hep-2 cells in various groups.Flow cytometry was also used to analyze the cell cycle progression of Hep-2 cells and apoptotic rate after administration.Results The proliferation inhibition rates in 3.0,6.0 and 12.0 mg?L-1 taxol groups were 14.0%,23.9% and 36.7%,respectively,there were significant differences between three groups(P

Objective To explore the effect of triptolide(TL) combined with hyperthermia on proliferation and apoptosis of Hep-2 cells and its affect on cell cycle phases,and provide theoretical foundation for treatment of laryngocarcinoma.Methods Hep-2 cells at logarithmic growth phase were randomly divided into control,hyperthermia,TL,TL combined with hyperthermia groups.MTT method was adopted to investigate the proliferation inhibition rate of the Hep-2 cells.Cell cycle and apoptotic rate of Hep-2 cells were analyzed by flow cytometry,and then the changes of subcellular structures were observed by electron microscope.Results The proliferation inhibition rates in hyperthermia,TL,TL combined with hyperthermia groups were 21.2%,28.5% and 54.5%,respectively;the proliferation inhibition rate in TL combined with hyperthermia group was higher than those in hyperthermia and TL groups(P

Objective To explore the changes of human mitochondrial COXI,ND1 and ND6 genes expression induced by ionizing irradiation.Methods Changes of human COXI,ND1 and ND6 gene expression were detected by RT-PCR and Real-time PCR 8 h after the irradiation in human lymphoblastoid cell lines,which were exposed to 1-10 Gy 60Co γ-rays.And the dose-effect relationships between expression changes of the genes and the doses were analyzed.The changes of these three genes expression were also analyzed at different post-radiation time-points between 0.5 h and 72 h after irradiation of 5 Gy in order to explore the time-effect.Results The expression of three genes COXI,ND1 and ND6,showed either the dose-effect or the time-effect after irradiation.The gene expression levels of three genes up-regulated generally and the peak change time-point was 4 h after irradiation.Conclusion Ionizing radiation,msht induce the changes of mitochondrial gene expression,and the gene expression level is up-regulated.

Objective To study the cell apoptosis and the dynamic expression and significance of apoptosis-related genes in graft veins. Methods A rat experimental model of autogenous graft vein was established by transplanting the right external jugular vein to infrarenal abdominal aorta in 100 Wistar rats. TUNEL and immunohistochemistry were used to detect the apoptosis, the expression of apoptosis related genes bcl 2 and bax in vascular smooth muscle cells (VSMCs) of graft veins. Results Within the 8 weeks after transplantation, the apoptotic VSMCs in the graft veins were much more than those in the control group with the apoptotic rate reaching the peak〔(28.5?16.6)%〕 on the 2nd week and dropping to (8.1?2.8)% during the 4th to 8th week. There was statistical difference compared to the control group 〔(0.5?0.2)%, P

Objective To investigate the dose response of S100A4 gene expression in the irradiated lymphoblastoid cells AHH-1 at different time points post irradiation.Methods AHH-1 cells was exposed to different doses(0,1,3,5,8,10,15 and 18 Gy)of 60Co γ-rays,and its mRNA levels of S100A4 was detected by reverse transcription PCR and real-time PCR at 4,8,12,24,48 and 72 h after irradiation.Results Within the range of applied doses,the level of S100A4 gene expression was upregulated with a good dose-response (R2 =0.79-0.93,P ＜ 0.05) and had obvious difference at different time points (F =8.91,P ＜ 0.01).Conclusion S100A4 gene expression at transcriptional level could be detected easily and had optimum dose-responses at certain time points after irradiation,and hence is applicable as a dosimeter.

Objective To explore whether a low dose of 60Co γ-rays could induce the adaptive response in the formation of nucleoplasmic bridges (NPB) in human peripheral blood lymphocytes,and if so,the range of the priming dose.Methods Human peripheral blood samples from healthy males were collected and irradiated with 0,20,50,75,100,150 and 200 mGy (dose-rate was 25 mGy/min) of 60Co γ-rays.After 6 h,the samples were irradiated with a challenge dose of 2 Gy (dose-rate was 1 Gy/min).The cytokinesis-block micronucleus (CBMN) assay was carried out to analyze the NPB and micronuclei (MN) formation in binucleated cells.Results Within the dose range of 0-200 mGy,the yields of NPB and MN increased with irradiation dose of γ-rays and the dose response of NPB followed with a linearquadratic equation of y =(1.5 × 10-4) x2-(5.67 × 10-3)x + 0.598 (R2 =0.893 8).Compared with the samples irradiated with 2 Gy alone,the yields of NPB and MN were significantly reduced when the samples were irradiated with a priming dose of 75-100 mGy before 2 Gy irradiation (U =2.66,2.97,3.96,5.89,P ＜0.05).The biggest decrease ratio of NPB yields approached to 43.2％ at the priming dose of 100 mGy.Conclusions Low doses in the range of 75-100 mGy of 60Co γ-rays could induce the adaptive response of NPB formation in human peripheral blood lymphocytes.

Objective To use three different methods in attempt to estimate the biological dose of the patient partially exposed to 192Ir source at5.7 accident in Nanjing,so as to provide dosimetric information for clinical remedy of exposed patients in the emergency of a nuclear accident.Methods Peripheral blood samples were collected on days 5 after exposure.The biological dose was estimated by the yields of dicentrics plus rings ( dic + r),cytokinesis-block micronuclei (CBMN) assay and nucleoplasmic bridge plus FHC (NPB + FHC).The homogeneity of radiation exposure was examined by Poisson distribution of dicentrics.Results By using three different methods,the whole body equivalent dose was dic + r estimated to be 1.51 Gy (95％ CI 1.40-1.61),1.47 Gy (95％ CI 1.36-1.60) by CBMN and 1.30 Gy (95％ CI 1.00-1.60) by NPB + FHC,respectively.A non-poisson distribution was also detected,suggesting partial body radiation exposure.Conclusions The estimated whole body equivalent dose ot a non-uniform radiation exposure was consistent with clinical diagnosis,suggesting that the yields ofdic + r,CBMN,as well as NPB + FHC,are efficient approaches to the estimation of biological doses.