Objective To construct secreting type human TRAIL(shTRAIL) gene vector pcDNA3.1-HRE/Egr1-shTRAIL mediated by hypoxia/radiation double sensitive promoter,and observe the effect of hypoxia and radiation on shTRAIL.Methods HRE upper and lower strands were gotten by chemical synthesis,double strands HRE was gotten by PCR;pMD19T-Egr1 was digested by SacⅠ and Hind Ⅲ,then Egr1 was obtained,pshuttle-shTRAIL was digested by Kpn Ⅰ and BamH Ⅰ,then shTRAIL was obtained;HRE/Egr1 double sensitive promoter mediated shTRAIL expression vector pcDNA3.1-HRE/Egr1-shTRAIL was constructed by gene recombination technique,it was identified correctlly by enzyme digestion,PCR and sequencing.A549 cells were divided into normal,hypoxia(0.1%),irradiation(6 Gy) and hypoxia + irradiation groups.Results After enzyme digestion by BamH Ⅰ and Sma Ⅰ,the fragments which lengthes were 1284 bp and 4 998 bp,2 292 bp and 3 990 bp were obtained;the vector was amplified by PCR with Egr1 and shTRAIL primer,the products which lengthes were 469 bp and 820 bp were obtained;pcDNA3.1-HRE/Egr1-shTRAIL was sequenced,the result was same to designed,this demonstrated that the construction was right.The vectors were transfected into A549 cells of adenocarcinoma of lung,the expression levels of shTRAIL mRNA and protein were increased after treated with hypoxia and radiation,it had statistically significant differences compared with normal group(P

OBJECTIVE:To explore the effects and safety of dexmedetomidine combined with remifentanil on postoperative cognition and hemodynamics in patients underwent colon cancer surgery.METHODS:One hundred undergoing colon cancer sur gery in our hospital during Jun.2013-Apr.2016 were selected and divided into control group and observation group according to random number table,with 50 cases in each group.Control group was given Remifentanil hydrochloride for injection 2-4 μg/kg for anesthesia induction,with maintenance dose of 0.5-2 μg· kg/min;observation group was treated with Dexmedetomidine hydrochloride for injection 0.5 μg/kg and remifentanil 2-4 μg/kg for anesthesia induction,with maintenance dose of Dexmedetomidine hydrochloride for injection 0.4 μg·kg/h+Remifentanil hydrochloride for injection 0.5-2 μg·kg/min.MMSE score and the incidence of postoperative cognitive dysfunction (POCD) were observed in 2 groups 1,2,3 d after surgery,and the occurrence of ADR was record ed.RESULTS:The incidence of POCD in observation group 1,2,3 d after surgery were 16.0％,4.0％,6.0％,which was signifi cantly higher than 36.0％,12.0％,10.0％ of control group,with statistical significance (P＜0.05).There was no statistical significance in MMSE score between 2 groups 1,3 d after operation (P＞0.05).2 d after surgery,MMSE score of observation group was significantly higher than that of control group,with statistical significance (P＜0.05).There was no statistical significance in hemodynamic indexes,the incidence of ADR as blood pressure increasing,amyostasia,nausea and vomiting between 2 groups 1,2,3 d after surgery (P＞0.05).CONCLUSIONS:Dexmedetomidine combined with remifentanil can significantly improve postoperative POCD in patients underwent colon cancer surgery and have little effect on hemodynamics with good safety.

Objective To study the security of low dose and half-body irradiation by 60Co ?-rays as a new method of clinical radiotherapy.Methods The simulated manikin was used to simulate human body and two radiation modalities of facing and backing on radioactive source were adopted.Half-body irradiation was done by 60Co ?-rays with doses of 9,10 and 11 cGy.The exposure dose of every layer and important target organs in the simulated manikin were detected,and the security of low dose and half-body irradiation as a therapeutic method was evaluated.Results The exposure dose of every layer and sensitive organs were all within safety margin,when simulated manikin facing or backing on the radioactive source was irradiated by 60Co ?-rays with doses of 9,10 and 11 cGy.Further,the exposure dose of sensitive organs in the simulated manikin backing on the radioactive source was lower than that in those facing the radioacive source.Conclusion The method of low dose and half-body irradiation as a radiotherapeutic method is safe and feasible and the radiation modality of backing on the radioactive source is more safe.

Objective To construct the recombinant plasmid that highly expressed human subcellular PTEN,in order to provide a basis for further study on its anti-tumor effect. Methods PTEN cDNA was amplified by RT-PCR based on mRNA of placenta.The PCR product was ligated into T-vector,and transfected into E.coli;the obtained T-PTEN plasmid was identified with restrictive digestion and sequencing.PCR was used to incorporte nuclear signal of localization(NSL) into PTEN when T-PTEN was used as template.Then the PCR product was ligated into T-vector,and transfected into E.coli,and T-NSL-PTEN plasmid was obtained.pcDNA3.1 and T-NSL-PTEN were ligated after digested with EcoRⅠand BamHⅠ,and transfected into E.coli,the recombinant vector pcDNA3.1-NSL-PTEN was obtained,and identified with digestion and sequcncing.Results The recombinant expression vector DUM-PTEN and PUM-NSL PTEN were identified by restrictive digestion and DNA sequencing.As expected,by EcoRⅠ and BamHⅠ digestion,it showed the band of 1 200 bp.The sequencing result showed the NSL was incorporated successfully.The recombinant pcDNA3.1-PTEN was obtained with 1 200 bp,the sequencing result showed that its sequence was same as target gene;the recombinant pcDNA3.1-NSL-PTEN was comfirmed by restrictive digestion and sequencing,and the NSL was incorporated successfully. Conclusion The recombinant expression plasmid pcDNA3.1-NSL-PTEN is constructed successfully which can highly express human subcellular PTEN.

Objective To evaluate the therapeutic effect of adipose-derived mesenchymal stem cells on radiation enteritis.Methods A total of 52 male Sprague-Dawley rats were used in the present study.Herein,46 rats were randomly selected and irradiated with a dose of 15 Gy at their abdomens.Two hours post-irradiation,23 rats were randomly selected and infused intraperitoneally with adipose-derived mesenchymal stem cells in passage 6 from young-female donor.The other 23 rats were intraperitoneally infused with PBS.The rest 6 rats were set as normal control.During the first 10 days post-irradiation,peripheral blood-samples from irradiated rats were harvested for testing the levels of IL-10 in serum using ELISA assay.Additionally,after isolating the thymic cells and peripheral blood mononuclear cells,the percentages of CD4/CD25/Foxp(3)-positive regulatory T cells in thymus and peripheral blood were tested by flow-cytometry.Finally,infiltration of inflammatory cells and deposition of collagens within irradiated small intestine were analyzed by H&E staining and Masson Trichrome staining,respectively.Based on the MPO-immunohistochemistry staining,the type of infiltrated cells was identified.The Kaplan-Meier method was used for analyzing the survival rate of irradiated rats.Results During a period of 30 days post-irradiation,the irradiated rats receiving adipose-derived mesenchymal stem cells survived longer than those receiving PBS (t =4.53,P ＜ 0.05).Compared to the irradiated rats with PBS-treatment,adipose-derived mesenchymal stem cells could elevate the level of IL-10 in serum (7 d:t =13.93,P ＜ 0.05) and increase the percentages of CD4/CD25/Foxp(3)-positive regulatory T cells in both peripheral blood (3.5 d:t =7.72,7 d:t=11.11,10 d:t =6.99,P ＜0.05) and thymus (7 d:t =16.17,10 d:t =12.12,P＜ 0.05).Moreover,infiltration of inflammatory cells and deposition of collagens within irradiated small intestine were mitigated by adipose-derived mesenchymal stem cells.Conclusions Adipose-derived mesenchymal stem cells were capable of curing radiation enteritis.

Objective To assess the therapeutic effect of human adipose-derived mesenchymal stem cells on radiation-induced vascular injury in the small intestine of rat. Methods A total of 34 male Sprague-Dawley rats were enrolled in this study. To establish a model of radiation-induced intestinal injury, each rat was irradiated with 15 Gy in whole abdomen. 17 rats were randomly selected and infused intraperitoneally with passage 6 ( P6 ) Ad-MSCs, and the other 17 rats that received PBS were set as control. 10 days post-irradiation, the number of CD31+ endothelial cells in the small intestine villus was measured by flow-cytometry, the expressions of CD31, CD105 and isolectin-B4 in the na?ve endothelial cells with detected by IHC-staining, and the vascular integrity was evaluated by measuring VE-Cadherin. The origination of na?ve endothelial cells within injured intestine was also analyzed. In addition, total mRNA were extracted from irradiated small intestine to assay the expressions of VEGF, bFGF, Flk-1 and SDF-1 using quantitative Real-time PCR. Results Compared to the control, the amount of CD31-postive endothelial cells within irradiated intestine was significantly increased after Ad-MSCs infusion ( t=12?15, P<0?05). The microvascular density in the injured sites was also significantly increased by the infusion of Ad-MSCs (20 d:t=10?33, P<0. 05;30 d:t=32?85, P<0?05). Moreover, the expressions of VEGF, bFGF, Flk-1 and SDF-1 were significantly up-regulated after delivery of Ad-MSCs ( VEGF:t =10?34, bFGF:t=11?25,Flk-1:t=6?73, SDF-1:t=6?73, all P<0?05), which was beneficial in maintaining the integrity of intra-villus blood-vessels as well as promoting neovascularization in the injured sites. Conclusion Ad-MSCs had potentials in healing radiation-induced vascular injury in rat small intestine.

Objective:To study effects of large dose thymopeptides on T cell subpopulations of patients with malignant tumor under radiotherapy.Methods:Fifty one patients with malignant tumor under radiotherapy were divided into 2 groups with 100 mg and 200 mg thymopeptides respectively.The patients we re given thymopeptides,100 mg/d or 200 mg/d,iv, for 10 days.The positive percent ages of CD4,CD8,CD25 and CD56 in T cells of peripheral blood before and after thymopeptide treatment were determined by flow cytometry.Results:The positive percentages of CD4 and CD25 in T cells of peripheral bl ood after 100 mg/d thymopeptide treatment were significantly higher than those befor e thymopeptide treatment (P<0.05),while those of CD4，CD8，CD25 and CD56 in T cells of peripheral blood after 200 mg/d thymopeptide treatment all increased significantly (P<0.05 or P<0.01). Conclusion:These results suggest that large dose of thymopeptides can increase i mmune function of patients with malignant tumor under radiotherapy,and the curat ive effect of 200 mg/d thymopeptides is better.

Objective To probe the influence of electron beam radiotherapy in different manners using different tissue equivalent boluses on skin and lung.Methods Adult female cavia cobayas were randomly divided into four groups as control group,fuU-time with bolus group,half-time with bolus group and without bolus group.Acute-irradiation animal models were established using electron beam in different manners with or without 0.5 cm tissue equivalent bolus.Pathological changes in lung,hair vesicle and fibroblast cell count were analyzed 40 clays after irradiation.Results The radiation dermatitis in the group with bolus was slighter than that of the group without bolus,but the radiation pneumonia was reverse.With bolus,the radiation dermatitis of haft-time group was slighter than that of full-time group.The injury repair of half-time group was more active than full-time group.Conclusions The treatment of haft-time bolus could protect lung without serious skin complications.

Objective To analyze the difference of dosimetry and evaluate clinical efficacy and acute toxicity reaction between RapidArc and fixed gantry angle dynamic intensity modulated radiotherapy (IMRT) in advanced cervical carcinoma.Methods A total of 43 patients with locally advanced cervical cancer were studied,including 22 patients treated with RapidArc and 21 patients with IMRT.All plans were prescribed 50.4 Gy in 28 fractions.The conformity index and homogeneity index of the targets,the monitor units(MUs) and delivery time were compared.Incidence of acute intestinal and bladder side effects and rates of efficacy were calculated.Results The conformity index of RapidArc was better compared to IMRT.The V40 and V50 of bladder and V30,V40 and V50 of rectum planned by RapidArc was significantly lower than that by IMRT(t =-2.386,-2.397,P ＜0.05;t =-5.525,-2.883,-2.686,P ＜0.05).The mean dose of femoral head planned by RapidArc was also significantly lower (t =-2.395,P ＜ 0.05).For RapidArc,mean MU and treatment time were reduced by 53.15％,and 62.14％,respectively.There was no difference in the incidence of acute intestinal and bladder toxicity and rates of complete remission and efficacy between the two groups.Conclusions In dosimetric analysis,RapidArc showed advantage in protecting organs at risk and reducing treatment time in radical radiotherapy for locally advanced cervical carcinoma.

Objective:To explore the dose distribution characteristics between RapidArc and five-field intensity modulated radiotherapy(5F-IMRT) plans in the treatment of locally advanced pancreatic carcinoma,and to provide reference for selecting the appropriate radiation technique in clinic.Methods:Ten patients with locally advanced pancreatic carcinoma were selected.The patients were scanned by simulation CT, and the targets and organs at risk were contoured. RapidArc plan and 5F-IMRT plan were designed respectively.The treatment time and the differences of dose distribution in the targets and organs at risk of RapidArc and 5F-IMRT plans were compared.Results:The conformal index (CI) of RapidArc plan was superior than that of 5F-IMRT plan(P=0.01).The homogeneity index (HI) of targets were similar between two plans (P>0.05).RapidArc plan decreased the maximum dose (Dmax) of the spinal cord(P=0.005);RapidArc plan decreased the mean dose(Dmean) of stomach(P=0.019);5F-IMRT plan decreased the V20 of kidney(P=0.043);RapidArc plan decreased the mean dose(Dmean) of small intestine(P=0.011).The small intestine V10 and V20 of RapidArc plan were lower than those of 5F-IMRT plan(P=0.015,P=0.14);the monitor unit (MU) of RapidArc plan was significantly lower than that of 5F-IMRT plan,with a 18% reduction from the MU level of the latter one,and the treatment time was reduced by 70.3%.RapidArc plan had the smaller doses at liver and kidney compared with 5F-IM RT plan.Conclusion:For the patients with locally advanced pancreatic carcinoma,RapidArc plan has higher CI. RapidArc plan shows the advantages in the protection of organs.Compared with 5F-IMRT,RapidArc plan has less treatment time and significantly improves the curative efficiency.In clinic, the RapidArc plan is recommended.