Objective To investigate the dynamic changes of VEGF mRNA and protein in the injured brain of rats irradiated by 20 Gy of electron beams.Methods A total of 114 Sprague-Dawley rats were randomly divided into two groups,42 rats in sham group with sham-irradiation,and the other 72 rats in irradiation group that the whole brains of rats were administered with a single dose of 20 Gy electron beam (6 MeV) to establish an animal model of radiation-induced brain injury.All rats were terminated at 1,3,7,14,28,42,56 d post-irradiation respectively.The pathological changes were observed with light microscopies.Real-time quantitative reverse transcriptase-PCR was used to measure the level of VEGF mRNA in the brain tissue.The expression of VEGF protein in whole brain tissue was detected with Western blot.The VEGF protein expression in brain endothelium,gliacyte and neuron were measured via immunohistoehemistry with computerized image processing.Results The pathological process of radiationinduced brain injury (RBI) included brain endothelial cell damage,brain edema,thrombosis formation and ablation,revascularization and angiogenesis.In the irradiation group,the expression of VEGF mRNA was declined significantly at 1,3,7,2g,42 d post-irradiation (t =16.275-46.118,P ＜ 0.05).The VEGF protein expression in whole brain tissue was up-regulated at 1 and 7 d after irradiation,but downregulated at 3,14,28,42,56 d post-irradiation.There were VEGF positive cells in the brain endothelium,gliacyte and neuron at 1 d after irradiation.The expression of VEGF protein in gliacyte and neuron significantly was increased at 1,14,42,56 d post-irradiation (t =-8.394--4.697,P ＜ 0.05),and increased significantly in brain endothelial cells at 1,14,and 42 d after irradiation (t =-5.554--4.159,P ＜ 0.05).Conclusions The expression of VEGF mRNA was relatively suppressed in RBI rats.The expression of VEGF protein in whole brain tissue was up-regulated only at the early stage of RBI.The expressions of VEGF protein in brain endothelium,gliacyte and neuron were increased in the observation period,which induced brain edema and the formation of thrombosis at the acute RBI stage,and participated in the progression of angiogenesis and thrombosis ablation at the early-delayed RBI stage.

Objective Based on the published articles,this paper aims to study the pattern and probability of lymph nodes metastasis in nasopharyngeal carcinoma (NPC) and establish a radiotherapy standard for selecting and delineation of clinical target volume of neck lymphatics for NPC patients with N0 or N1.Methods Clinical trials about NPC by MRI as the main or only diagnostic imaging way were collected and extracted from database,such as WanFang,CBM,PubMed,Cochrane Library.Statistical analyses were performed using Stata software,version 12.0.Results Twenty-one clinical trials from 963 original articles were included in this analysis.The most commonly involved regions included retropharyngeal (67％) and level Ⅱ lymph nodes (71％).The overall probability of levels Ⅲ,Ⅳ and Ⅴnodal involvement were 37％,14％ and 21％,respectively.Low-risk node groups included levels ⅠA,ⅠB,Ⅵ nodes,and the rates of lymph node metastasis were 0,2％,2％ and 1％,respectively.Nodal metastases followed an orderly pattern of top-down,and the probability of skip metastasis between levels varied between 0.5％ ～7.9％.Conclusion Lymph node metastasis in NPC follows a predictable and orderly pattern.The rarity of metastasis in certain nodal groups andskip metastasis suggest that reduced treatment volume is feasible in conformal radiotherapy for NPC patients with No or N1.

Objective To evaluate the effect of cell-penetrating peptide (protein transduction domain 4,PTD4) mediated copper-zinc superoxide dismutase (Cu/Zn SOD) on hypoxia/reoxygenation injury (HRI) in rat myocardial cells.Methods Rat myocardial cell H9C2 HRI model was prepared by using the anaerobic incubator (85% N2,10% H2,5% CO2).The HRI group (without adding any treating factor in HRI cell culture fluid),HRI+Cu/Zn SOD group (adding 10 μmol/L Cu/Zn SOD) and HRI+PTD4-Cu/Zn SOD group (10 μmol/L PTD4-Cu/Zn SOD) were set up.In addition,normally cultured myocardial cells served as the normal control group.After incubating for 30 min,the ultra microstructure of mitochondria was observed under transmission electron microscope.The mitochondrial membrane potential was detected by JC-1 kit.The myocardial cell apoptosis was detected by TdT mediated dUTP nick end labeling TUNEL technique.Results The mitochondria injury degree after 30 min incubation in the PTD4-Cu/Zn SOD group was significantly improved compared with the HRI group.Compared with the normal control group,the mitochondrial membrane potential in the HRI group was significantly decreased,while the mitochondrial membrane potential in the PTD4-Cu/Zn SOD group was lower than that in the normal control group,but compared with the HRI group,which was obviously recovered.The cardiomyocyte apoptosis in the HRI+PTD4-Cu/Zn SOD group was (10.20±2.77)%,which was significantly decreased compared with (28.40±2.41)% in the HRI group,the difference was statistically significant (P<0.05).Conclusion PTD4 mediated Cu/Zn SOD can attenuate HRI in rat myocardial cells.

Objective To describe the clinical characteristics,outcome and prognostic factors of primary intraspinal tumor treated with postoperative radiotherapy.Methods Thirty-three patients diagnosed as suffering from intraspinal tumor treated with postoperative radiotherapy were analyzed.There were eighteen male and fifteen female,with a median age of 37 years(range 7-64 years).Two patients were treated with grossly total resection,25 subtotal resection,2 biopsy only.These patients were treated either with(~(60)Co)(n =18,before 1998) or 6?MV-X ray(n =15,after 1998) to the total dose of 45-68?Gy.The overall response and survival rates were calculated by Kaplan-Meier method and the difference between groups was tested by the log-rank method.The prognostic factors related to the characteristics of patient,tumor,and treatment were determined.Results The overall response,five-year and disease-free survival rate was 81.3%,59.4% and 56.3%,respectively.By multivariate analysis,the histological type(P= 0.039) and gender(P= 0.049) had independent impact on the 5-year survival rate.Ependymoma had a better prognosis compared to other histological subsets and female gained more survival advantage over male.There was no close relationship between survival and other factors,such as age,extent of surgery,initial site,irradiation dose,or the interval between surgery and radiotherapy.Conclusions Postoperative radiotherapy is safe and effective for primary intraspinal tumor.The histological type and gender are important prognostic factors of survival in primary intraspinal tumors.

Objective To investigate the dynamic changes of nuclear factor-κB(NF-κB) during brain injury of irradiated rats.Methods According to the random number table method,82 healthy Sprague-Dawley rats were divided into two groups:50 rats in the irradiation group and the other 32 rats in the control group given sham-irradiation.The whole brains of rats in the irradiation group were irradiated with a single dose of 20 Gy electron beam to establish an animal model of radiation-induced brain injury.All brain tissues were respectively taken out 1,3,7,14 and 28 d after irradiation.The quantificational real-time-polymerase chain reaction (QRT-PCR) was used to detect the expressions of the NF-κB mRNA while the expression of NF-κB protein was analyzed by ELISA and Western blot.The positioning expression of NF-κB protein in the hippocampus was measured with immunohistochemical staining.Results In the irradiation group,the expression of NF-κB mRNA significantly increased at 3 and 7 d following irradiation (t =37.79,35.30,P ＜ 0.05) and it approached a peak value on the third day following irradiation.The expression of NF-κB protein in whole brain tissues after irradiation was up-regulated and reached a peak on the first day post-irradiation and it was significantly higher than controls at 1,3,7,14 d after irradiation (t =30.94,14.87,27.17,13.27,P ＜ 0.05) then declined and dropped to the normal level by 28 d.The results of immunohistochemical staining showed that the number of NF-κB positive cells in the hippocampus increased at 1,3 and 7 d following irradiation (t =-8.49,-4.47,-3.46,P ＜ 0.05).Conclusions After irradiation,the expressions of NF-κB mRNA and corresponding protein first increase and then later decrease,which promotes the development of brain injury.

Objective:To evaluate the knowledge and recognition of the influence factors of adverse drug reactions in medical staffs in Nanjing.Method:A structured questionnaire was used to collect information on ADR related knowledge in medical staffs in some selected hospitals in Nanjing.Result:972 valid questionnaires were obtained form 1200 selected subjects,and the response rate was 81.0%.The scores were significantly different in age groups,sex and different kinds of occupational medical workers.The mean levels of the scores were analyzed between different groups.The single factor analysis demonstrated that the different levels of hospitals,sex,knowledge and occupation were correlated with the recognition. The multiple factors logistic regression analysis demonstrated that the recognition of adverse drug reaction among medical staff in Nanjing was related with different levels of hospitals and different occupations.Conclusion:In order to enhance the recognition level,it is necessary to carry out both the long-term education and train about the specialized knowledge of ADR in basic hospitals and non-pharmacy medical staff.

Objective　To investigate the association betwee n the β3-adrenergic receptor gene (β3-AR) and obesity. Methods　147 patients with obesity and 106 controls were selected. Genotype of the β 3-AR was determined by polymerase chain reaction (PCR) and restriction fragme nt length polymorphism methods (RFLP). Body mass index (BMI) was detected. Results　The frequency of Arg64 allele of β3-AR gene in the obesit y g roup (20.3%) was significantly higher than those in the control group (10.2%) ( P=0.001) . The distribution of Trp64Arg and Arg64Arg genotype in the patient group (38.7% and 0.9%) were both higher than that in the control group (19.0% and 0.7%). In the female subj ects, the Trp64Arg mutation and frequency of Arg64 allele of the β3-AR gene in the obesity group(40.6% and 20.3%) were also higher than those in control (16 . 7% and 9.5%)(P<0.01). Conclusions　These results suggest th at the Trp64Arg mutation of the β3-adrenergic-receptor gene may be associat ed with increased body fat.

Objective To investigate the radiosensitizing effect of knock-down the expression of miR-21 on human SHG-44 glioma cells and explore the possible mechanism. Methods Antisense oligonuleotidas of miR-21, mediated by LipofectamineTM 2000, were transfected to SHG-44 cells. Three groups were: blank control group ( mock group), negative control and antisense transfected group ( AS-miR-21 gorup). Cells of each group were irradiated with 6 MeV X-rays at the doses of 0,1,2,4,6 and 8 Gy.Dose-suvivial curve was established by colony-forming assay. The influence of AS-miR-21 on cell cycle and cell apoptosis was analyzed by flow cytometry assay after 6 Gy irradiation. Results The value of D0 and Dq of AS-miR-21 group declined obviously compared with the mock group and negative control group. Flow cytometric analysis showed that cell cycle distribution changed( G0/G1 phase arrest, S phase decreased)after transfected with AS-miR-21 (t = 8.18, -4.52,P ＜ 0.05 ). The sensitization enhancement ratios of D0 and Dq were 1.32 and 2.10 respectively. Apoptosis assay showed the early apoptosis rate was signiflcantely increased in AS-miR-21 、irradition alone and combined group than mock control group( t = 20.14,11.11,50.07, P ＜ 0.05). Conclusions AS-miR-21 can enhance the radiosensitivity of human glioma cells SHG-44 by promoting cell apoptosis and faciliating cell cycle redistribution.

Objective To investigate the effects of radiosensitivity enhancement and inhibition of migration ability of human lung adenocarcinoma cells by celecoxib,a selective cyclooxygenase (COX)-2 inhibitor.Methods Human lung adenocarcinoma cells of the line A549 were cultured and then inoculated into six-well plates and randomly divided into 4 groups:control group,celecoxib group administered with celecoxib at the subtoxic doses 30 and 50 μmol/L,irradiated group exposed to 0,1,2,4,6,or 8 Gy by linear accelerator,and combined treatment (celecoxib + irradiation) group.The radiosensitizing effect of celecoxib was assessed by clonogenic cell survival test.The migration ability of the A549 cells was measured by scratch-wound test and the content of metalloproteinase-2 (MMP-2) in culture supernatant was detected with ELISA.Results The sensitization enhancement ratio of the celexib group was increased dosedependently.The values of D0 ,Dq,SF2 and D0.01 of the celecoxib + irradiation group were all significantly lower than those of the irradiated group.Scratch-wound test showed that the no-scratch area of the celecoxib + irradiation group and celecoxib group were all significantly wider than those of the mere irradiation and control groups and there was a dose-dependent manner,and the no-scratch area of the celecoxib + irradiation group was wlider than that of the celecoxib group.ELISA showed that the MMP-2 levels in the supernatant of the celecoxib group and celecoxib + irradiation group were respectively significantly lower than those of the control group and mere irradiated group (t = 3.78,5.79、3.15,P ＜ 0.05),however,there was not significant difference between the mere irradiation and control groups (t = 2.73,2.38,P ＞ 0.05).Conclusions Celecoxib enhances concentration-dependently the radiosensitivity of human lung carcinoma cell and inhibits the secretion of MMP-2 of the carcinoma cells,thus inhibiting their migration ability.