Objective The techniques and early effect of perendoscopic intrasphincteric injection of botulinum toxin (BTXA) for achalasia was introduced and evaluated. Methods 13 patients with achalasia were enrolled in the study.Symptom scoring ( modified ) and esophagography ( measuring cardiac opening,height and width of contrast media retention in 5 minutes ) were performed before and one week after treatment.1 ml of BTXA (20 u) was injected endoscopically at a point of every quadrant (4 points) situated 0.5 cm above the dentate line into the lower esophageal sphincter. Results The symptoms improved remarkably the next day as chest pain,frequency and seriousness of dysphagia and regurgitation declined significantly (P

Objective To evaluate the efficacy of stereotactic body radiation therapy (SBRT) for hepatic metastases from colorectal cancer,and to collect data for the application of this technique.Methods A total of 28 patients from No.306 Hospital of PLA,including 17 male and 11 female with median age of 63.8 (range from 31 to 86),were treated with SBRT for colorectal cancer with liver metastases with 54 lesions in total.The GTV,CTV and PTV were delineated above the enhanced CT scans acquired during normal quiet respiration.CTV was obtained by adding 5 mm isotropic margin from GTV,and PTV was obtained by adding 5 to 10 mm isotropic margin from CTV.Prescription dose line covered 50％-60％ of isodose curve at 3-6 Gy/fraction.The total dose was 39-45 Gy and the biologically equivalent doses(BED)was 50.7-65.3 Gy.The patients were followed-up beginning at 3 months after SBRT.The change in size of the lesion based on enhanced CT or MR scans was evaluated.Toxicity was evaluated and scored according to the RTOG criteria.Local control rate and survival rate were analysed.Results All patients completed the treatment.With median follow-up of 15.1 months (range frome 3 to 30 months),7 patients survived at the end of follow-up.The local control rate (LC) was 79.2％,and 1-and 2-year overall survival rate(OS) were 82.7％ and 48.6％,respectively.There was a close corelation between the size of lesion and the LC.The LC (PR + CR) was much better at the size of lesion less than 14 cm3 than that at the size more than 65 cm3(x2 =4.17,P＜0.05).When the size was more than 180 cm3,the LC was zero.Toxicity included fatigue (60.7％),grade 1 and 2 digestive system toxicity (28.6％),a transient grade 1 and 2 bone marrow suppression (46.4％) and a transient increase in transaminase(17.8％).No grade 3 toxicity and above and late toxicity were observed.Conclusions Stereotactic body radiation therapy could be suggested as the first choice for the selected patients who suffer form colorectal liver metastases,especially for those who cannot undergo surgery.

Objective To evaluate clinical effect of volume modulated arc therapy ( VMAT) for spinal metastases .Methods Twenty patients with spinal metastases who had been treated with VMAT were chosen to participate in the study .The prescribed dose varied from 45 to 60 Gy within 15 -20 fractions, 3 Gy per fraction, and one fraction per day of VMAT .Pain and quality of life were measured before VMAT and at several time points up to 6 months after treatment , by the numerical rating scale (NRS) and verbal rating scale (VRS) and the quality of life scale for bone metastases (EORTC QLQ-BM22) questionnaire.In addition, Frankel grading was used to evaluate the neurological function of spinal cord.The primary endpoint was frequency and duration of complete pain relief , and the secondary endpoint was death.Results At the end of the follow-up, the number of patients reporting no pain from spinal metastases, as measured by the NRS, increased from 0 of 20 before VMAT to 10 of 14 ( t =20.24, P<0.05) .NRS-reported pain reduction from baseline to 1 weeks and 4 weeks after VMAT were clinically meaningful with NRS score (6.50 ±0.51) at baseline,(4.30 ±0.47)at 1 week and(2.50 ±0.50)at 4 weeks (t=15.98, 27.57, P<0.05).The pain reduction maintained until the end of the follow-up.These improvements were accompanied by significant reduction in opioid use during the first 6 months after VMAT (16 of 20 patients with strong opioid use at baseline vs.6 of 18 at 6 months; t =8.46, P <0.05). According to EORTC QLQ-BM22 questionnaire , patients obtained significant improvement in pain location , severity, and function with the score (46.50 ±1.50)at baseline, (38.35 ±0.98) at 4 weeks(t=21.51, P<0.05).However,no significant alleviation in social psychology was reported (P>0.05).After VMAT, the patients who suffered from nerve function impairment recovered to different extent . No one had radiation-induced injury , such as radiation myelitis , radiation pneumonitis , etc.Median survival time was 10 month.Conclusions VMAT is a safe and effective treatment method for spinal metastases .Significant reductions in patient-reported pain were observed , along with nerve dysfunction improved .The patients′quality of life was significantly improved .VMAT has no late spinal cord toxicities .

Objective To explore the effects of different dose rates of X-ray under the same dose on cell clonogenic formation in non-small-cell lung cancer cell line A549 in order to provide experimental basis for clinical radiotherapy plan. Methods The A549 cells were cultured at low density and irradiated with X-rays at dose of 4 Gy and selected dose rates of 1, 2, 4 and 6 Gy/min, respectively, from a linear accelerator. The 8th day after irradiation, the cells were fixed and stained with Giemsa solution, and colonies containing at least 50 cells were counted. The plating efficiency and surviving fraction were calculated. Results The clonogenic number in non-irradiated cells was 88.6±4.6. The numbers were significantly reduced in irradiated cells at dose rate 1, 2, 4 and 6 Gy/min (12.3±3.4, 9.0±0.8, 5.6±1.0, 11.5±1.7, respectively) than that in non-irradiated control cells (F=678.799, P<0.05). The plating efficiencies were decreased in irradiated cells, especially in 4 Gy/min irradiated cells, which was lower than that in any of the other three dose rate groups (P< 0.05). Conclusions Though at same radiation dose, cancer cells have different clonogenic formation efficiency when irradiation with X-ray at different dose rates. Thus, treatment with optimal dose rate may improve the radiotherapy efficacy.

Objective To explore the effect of dose rate of X-rays on migration of non-small cell lung cancer (NSCLC) cells and provide the experimental basis for developing radiotherapy scheme. Methods Human NSCLC cell line A549 was cultured and irradiated with X-rays at dose of 6 Gy from a linear accelerator. The dose rates of 1, 2, 4 and 6 Gy/min were selected. Monolayer adherent cells were scratched and photographed at 0 hour and 24 hours under a microscope to measure the scratch width. Results After 24 hours, the scratch width of nonirradiated control cells was (640.7±8.1)μm. The scratch widths of cells were different when cells were irradiated with X-rays of various dose rates. Scratch widths were the largest in cells irradiated at dose rates of 1 Gy/min [(691.4±7.6)μm] and 6 Gy/min [(691.8±12.1)μm]. The scratch width was (666.2±1.3) μm of X-rays at 4 Gy/min, and there were significant differences compared with nonirradiated group (all P< 0.01), which suggested that inhibitory effect of X-rays at dose rates on A549 cell migration was obvious. However, the scratch width of cells irradiated at 2 Gy/min [(643.5 ±6.8) μm] had no difference compared with the control cells (t=-0.336, P=0.742). Conclusions The effect of X-rays irradiation on cell migration of human NSCLC cell line A549 is related with irradiated dose rate. The effect of different dose rates on cell migration is significantly different. Selecting appropriate dose rates for irradiation may help to improve the efficacy of radiotherapy.

Objective To evaluate the therapeutic efficacy of stereotactic body radiotherapy (SBRT) with gamma knife on stage Ⅰ-Ⅱ non-small-cell lung cancer(NSCLC)and the quality of life of the patients undergoing this therapy.Methods Twenty NSCLC patients with the median age of 76,10 at stage Ⅰ and 10 at stage Ⅱ who were unable or unwilling to undergo surgery were given SBRT with gamma knife at the doses of 3-6 Gy in 8-15 fractions,finished within 2 to 3 weeks.The prescription isodose line was 50％,the marginal dose was 39-56 Gy,the central dose was 78-112 Gy,and the total biologically effective dose was 51-83 Gy.The patients were observed after admission and followed up by chest CT 1,3,6,and 12 months after treatment until progressive disease or death.EORTC QLQ-LC43 questionnaire was used to investigate the changes in quality of life.Results The 20 patients were followed up for 24 (12-46) months.At six months after the treatment,the overall response rate was 80％,and the complete response rate was 35％.The 1,2 and 3-year local control rates were 100％,95％ and 95％,respectively.The 1,2 and 3-year overall survival rates were 95％,80％ and 50％ respectively; The 1,2,and 3-year progression free survival rates were 85％,64％ and 33％,respectively.The failure rate was 20％ and the rate of progress within the planning target volume was 5％.No acute toxicity at grade 3 and over occurred in any patient during the treatment.15％ of the patients developed grade 1-2 radiation pneumonia.Age,gender,pathologic index or not were weakly correlated with the overall survival.The emotional function was improved significantly after treatment (P ＜ 0.05),dyspnea and cough were improved at different degrees,however,not significantly.There were no significant changes in the physical function and symptoms,such as fatigue,lack of appetite,insomnia,etc.Conclusions Significantly improving the motional function and maintaining the quality of life,SBRT with gamma knife is effective for elderly NSCLC patients with high local control rate fair overall survival rate and few side effects.

Objective To analyze the efficacy and prognosis of stereotactic radiotherapy (SRT) and whole-brain radiotherapy (WBRT) in treatment of brain metastases,and to observe the influence of temozolomide (TMZ) on survival rate during the period of radiotherapy.Methods A total of 52 patients with brain metastases were divided into two groups according to treatment methods,including 35 patients treated with WBRT plus SRT and 17 patients treated with SRT alone.WBRT dose was 1.8 - 3.0 Gy per fraction,one fraction a day,five fractions per week,with total dose of 30 - 40 Gy.After WBRT,gamma knife was performed with prescription isodose line of 45％ -70％ surrounding the planned target volume in WBRT + SRT group.The marginal dose was 12 - 15 Gy and the center dose was 20-30 Gy.In SRT group,the prescription isodosc line was 45％ - 70％ and the marginal dose was 36 - 40 Gy while the center up to 70 - 80 Gy.The follow up time was 1 - 2 years.Besides 20 patients in this study took temozolomide capsule during and after radiotherapy.The schedule of concomitant chemotherapy was temozolomide of 75 mg/m2 by oral administration every day until radiotherapy was over,and then temozolomide of 150 mg/m2 was taken for 3 -6 months after radiotherapy.Results The efficiency during 1 -3 months after treatment was 84.62％ in this study.In the WBRT + SRT group,the efficiency was 88.57％ and declined to 76.47％ in the SRT group.The six month-and one year-local control rate were 92.10％ and 85.20％,respectively.The average survival time of WBRT + SRT was 13.2 months and median survival time was 11 months.Six month-,one year-and eighteen months-survival rate were 71.40％,54.30％ and 14.30％,respectively.In the SRT group,the average survival time was 10.2 months and median survival time was 9 months.Six month-,one year- and eighteen month-survival rate were 41.20％,23.50％ and 5.88％,respectively,while those for RT + TMZ group were 80.00％,60.00％ and 10.00％.In comparison,those in RT group were 56.30％,37.50％ and 12.50％,respectively.Conclusions Effect of gamma knife stereotactic radiotherapy combined with WBRT is better than GK stereotactic radiotherapy alone in treatment of brain metastases.Compared with radiotherapy alone,concomitant temozolomide chemotherapy could improve the survival rate of the patients with brain metastases without increasirg adverse reactions significantly.

Polyethylenimine (PEI) is one of the most characterized non-viral vectors. It can condense DNA in a good manner and achieve high transfection efficiency. Minicircle DNA (mc-DNA) is a novel kind of supercoiled DNA which is devoid of bacterial backbone. mc-DNA is superior to conventional DNA for its higher transfection efficiency and longer time-span. In this study, we combined PEI and mc-DNA in gene delivery system. We investigated the physicochemical and biochemical effects of this non-viral system and further explore its potential in tumor gene therapy. mc-DNA was obtained by recombination of parental plasmid in the presence of L-arabinose, and complexed with PEI. The results of transmission electron microscopy and scanning electron microscopy showed that the particles were spherical and homogeneous. Through gel retardation assay and MTT assay, we found that there were no obvious differences in binding capability of PEI to mc-DNA and plasmid DNA, as well as in cytotoxicity. The results of dynamic light scattering showed that the size of PEI/mc-DNA was about 68 nm, a slight larger than that of PEI/plasmid DNA. Furthermore, the tumor cells transfected with mc-GFP showed higher GFP expression level than that of conventional plasmid. The same results were achieved in the cells treated with tumor-suppressor gene pten, assayed by RT-PCR and Western blot. It indicates that the system of PEI/minicircle DNA is promising in gene transfer.
DNA, Circular ; genetics ; Gene Transfer Techniques ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; genetics ; Nanoparticles ; chemistry ; PTEN Phosphohydrolase ; genetics ; Polyethyleneimine ; metabolism ; Transfection

Objective To investigate the effects of cerium oxide nanoparticles of different sizes on the number and constructions of immune cells in peripheral blood of mice after X-ray irradiation. Methods Mice were randomly divided into 4 groups according to body weight layer and the weight of each mouse was weighed. All mice were divided into 6 groups according to weight from high to low, and there were 4 mice in each group. Then 1 mouse was randomly taken from each group to form the control group. Model group, 5 nm and 25 nm cerium oxide nanoparticles groups were formed in turn. There were 6 mice in each group. The mice in model group and cerium oxide nanoparticles administration groups were irradiated once with 3 Gy of X-rays. The mice in cerium oxide nanoparticles groups began to be intraperitoneally administrated once a day with 10 μg 5 nm or 25 nm cerium oxide nanoparticles per kilogram body weight on the 4th day before irradiation and once every other 2 days after irradiation. The mice in the control group and model group were intraperitoneally administrated with 0.9 % saline. The mice were killed on the 10th days after irradiation. White cells count (WBC) and classification in peripheral blood were detected by using automatic globulimeter, and lymphocyte subsets were analyzed by using flow cytometry. Results Compared with the control group, the number of WBC, neutrophil granulocytes, monocytes, lymphocytes, total T lymphocytes, CD4+and CD8+T lymphocytes and the percentages in the model group were decreased (all P＜0.05), and percentages of the lymphocytes, B cells and NK cells and ratio of CD4 to CD8 were increased in model group (all P＜ 0.05). Compared with the model group, the above parameters except percentages of T lymphocytes, CD4+and CD8+T lymphocytes were improved in mice of 5 nm cerium oxide nanoparticle group (all P ＜0.05). Compared with the control group, the number of WBC and lymphocytes were decreased in the 5 nm cerium oxide nanoparticle group (P＜0.05), and there were no significances in other parameters compared with the control group (all P ＞0.05). Compared with the control group, the number of WBC and lymphocytes, the number and percentages of T lymphocytes, CD4+and CD8+T lymphocytes and the percentages were decreased (all P＜ 0.05), and percentage of NK cells and ratio of CD4 to CD8 were significantly increased in 25 nm cerium oxide nanoparticles group (all P＜ 0.05). The number of lymphocytes and CD8+T lymphocytes in 25 nm cerium oxide nanoparticles group was lower than that in 5 nm cerium oxide nanoparticles group (all P ＜ 0.05). Conclusions The effects of cerium oxide nanoparticles of different sizes on the immune cells of mice after X-ray irradiation are different, and 5 nm cerium oxide nanoparticle is superior to 25 nm cerium oxide nanoparticle.