Objective To apply RNA interference technique for reducing the expression of MDC1 gene in esophageal carcinoma cell line ECA109, observe the changes in cell cycle and radiosensitivity after radiation, and discuss related mechanisms. Methods Three pairs of effective interference sequences and negative control sequences were synthesized for MDC1 mRNA sequence, and a recombinant plasmid was constructed with the vector pSIH1?H1?copGFP. RT?PCR and Western blot were used to determine the expression levels of MDC1 mRNA and protein. Colony?forming assay was applied to measure radiosensitivity, flow cytometry to determine cell cycle, Western blot to determine the expression of CHK1 and CHK2 proteins, and laser scanning confocal microscope to observe the number of MDC1 blotches inside the nucleus. One?way analysis of variance was used to analyze the differences between groups. Results The pSIH1?H1?copGFP plasmid was constructed successfully and ECA109 cells were infected to obtain ECA109M cells with stable transfection. The expression levels of MDC1 mRNA and protein in ECA109M cells were lower than those in ECA109N and ECA109 cells ( P= 0. 032 and 0. 041, respectively ) . After 5?Gy radiation, ECA109M cells had a lower proportion of G2+M cells than ECA109N and ECA109 cells ( P=0. 026) . After 5?Gy radiation, ECA109, ECA109N, and ECA109M cells had similar expression levels of CHK1 and CHK2 proteins ( P= 0. 345 and 0. 451, respectively ) , and ECA109M cells had a lower expression level of CHK2 T68 protein than ECA109 and ECA109N cells ( P=0. 012) . ECA109 cells had a D0 value of 3. 06 Gy and an SF2 value of 0. 91;the D0 values for ECA109N and ECA109M cells were 2. 90 Gy and 1. 88 Gy, respectively, and the SF2 values for them were 0. 89 and 0. 84, respectively ( P=0. 021 and 0. 037, respectively ) . Conclusions RNA interference can reduce the expression levels of MDC1 protein and cell cycle?related proteins, release cell cycle arrest, and enhance radiosensitivity in esophageal carcinoma ECA109 cells.

Objective To investigate the effects of inhibition of MDC1 protein expression on xenografted tumors in nude mice,and to observe the histopathological and cellular changes in nude mice.Methods Three pairs of effective and control short hairpin RNA targeting MDC1 mRNA were designed and cloned into the pSIH1-H1-copGFP vector.Real-time PCR and Western blot were used to determine the mRNA and protein expression of MDC1.After selection by copGFP reporter gene,cells were divided into negative transfection group (ECA109-N) and MDC1 transfection group (ECA109-M).The transfected cells were injected into nude mice.The mice were divided into ECA109 group,ECA109-N group,and ECA109-M group.Each group was divided into irradiation subgroup and non-irradiation subgroup.The changes in tumor size after irradiation were evaluated in each group.Western blot was used to measure the expression of CHK1,CHK2,and CHK2T68 in xenografted tumors.Flow cytometry was used to analyze the cell cycle distribution and apoptosis of tumor cells in nude mice.The variance analysis was used to compare the mean of multiple groups,and the SNK-q test was used in the two two groups.Results The pMDC1-shRNA plasmid was successfully constructed and used to transfect ECA109 cells.ECA109-M cells were obtained by stable transfection with the recombinant plasmid.All inoculated nude mice survived with visible xenografted tumors at the underside of the paw in about one week.There was no swelling and wound in inoculation sites.There was no significant difference in tumor size between different groups (P＞0.05).The tumor growth in the ECA109 group and the ECA109-N group significantly slowed down after irradiation with a dose of 15 Gy (P＜0.05).Compared with the other two groups,the ECA109-M group had a significant smaller tumor size,significantly slower relative tumor growth,and significantly higher growth inhibition (all P＜0.05).The q value of the ECA109-M group was 1.36.In the ECA109-M group,there were no significant changes in the protein expression of CHK1 and CHK2 after irradiation (P＞ 0.05);however,the phosphorylation of CHK2T68 protein was significantly reduced after irradiation (P＜0.05).There were no significant differences in cell cycle distribution or the proportion of apoptotic cells in tumor tissue between the three groups (P＞0.05).Conclusions Inhibition of MDC1 protein expression by RNA interference can effectively inhibit the growth of xenografted tumors after irradiation in the nude mice by increasing their radiosensitivity.

Objective: To identify the relationship between the expression of protein tyrosine phosphatase non-receptor type 12 (PTPN12) and radiotherapy effect in non-small cell lung cancer (NSCLC) tissues and to determine whether PTPN12 deficiency can sensi-tize lung cancer cells to irradiation. Methods: From September 2013 to October 2014, 92 NSCLC patients undergoing radiotherapy with or without platinum-based combination chemotherapy were analyzed retrospectively. Before the treatment, PTPN12 expression was detected through immunohistochemistry. After the completion of radiotherapy, the patients' responses were assessed and radio-therapeutic efficacy analyzed. The human NSCLC cell line H1299 was infected with shPTPN12 knockdown, and colony survival assay was analyzed after irradiation. Chi-square test was used to examine the correlation between PTPN12 expression and clinicopathologi-cal characteristics. Univariate analyses and Logistic regression test were used to analyze the relationship between clinicopathological characteristics and radiotherapeutic response. Results: Patients with low PTPN12 expression were more sensitive to radiotherapy than those with high PTPN12 expression (80.0%vs. 57.1%, P=0.018). Multivariate analysis showed that PTPN12 expression was the on-ly independent predictor of radiotherapeutic response in NSCLC. The H1299-shPTPN12-knockdown cells were sensitive to irradiation. Conclusions:The results of the study indicated that downregulation of PTPN12 improved the radiosensitivity of NSCLC cells.

Objective To investigate the possibility and accuracy of predicting malignant middle cerebral artery infarction (mMCAI) with bedside electroencephalography (EEG). Methods Thirty-five patients with massive hemispheric infarction (MHI) underwent bedside EEG monitoring within 48 h of onset. The EEG indicators were interpreted blindly, and the clinical, laboratory and imaging parameters were analyzed. The patients were divided into mMCAI group and non-mMCAI group according to whether they had occurred mMCAI or not within 7 days of onset. The differences of EEG indicators, clinical, laboratory and imaging parameters between the 2 groups were compared. When the parameters of significant difference and statistical significance appeared the odds ratio (OR) of occurring mMCAI were analyzed, and their accuracy of predicting mMCAI was calculated. Results Of the 35 patients with MHI, 20 were in the mMCAI group and 15 were in the non-mMCAI group. There were significant differences in the EEG indicators (infarction on the contralateral side, including disintegration of occipital α rhythm, generalized slow-wave, dominant frequency wave low amplitude, regional attenuation without delta [RAWOD]pattern, and absence of EEG reactivity), clinical parameters (nausea accompanied with vomiting), and imaging parameters (the infracted area more than the entire MCA territory, and midline shifting 3 to 5 mm at the level of septum pellucidum) between the 2 groups (P < 0. 05). Of those, the risk of mMCAI was the highest in patients with disintegration of occipital a rhythm on the contralateral side of infarction (P = 22. 67, 95% CI 3. 89-132. 10). The sensitivity of predicting mMCAI was 85. 0%, the specificity was 80.0%, the positive predictive value was 85.0%, and the negative predictive value was 80. 0%, which were superior to other EEG indicators and clinical or imaging parameters. Conclusions Bedside EEG indicators can early predict mMCAI, moreover, the predictive accuracy is superior to the clinical and imaging parameters.

in order to maintain a normal temperature range.

Objective To investigate the value of prophylactic irradiation to the lymphatic drainage area in radical three-dimensional conformal radiotherapy (3DCRT) and to evaluate the efficacy and adverse effects of 3DCRT with different clinical target volumes.Methods A retrospective analysis was performed on the records of 219 esophageal cancer patients without distant metastasis who received 3DCRT from January 2005 to December 2010.One hundred and five patients received involved-field irradiation (IFI) with a total dose of 54-66 Gy;114 patients received elective nodal irradiation (ENI) with a total dose of 46-52 Gy; the prescribed dose to the primary lesion was 56-70 Gy.The Kaplan-Meier method was used to calculate local control (LC) and overall survival (OS) rates,and the log-rank test was used for univariate prognostic analysis.Results The 1-,3-,and 5-year sample sizes were 219,172 and 67,respectively.The 1-,3-,and 5-year LC rates for IFI group were 63.0％,39.1％,and 27.2％,respectively,versus 70.5％,53.3％,and 51.7％ for ENI group (x2 =6.22,P =0.013) ;the 1-,3-,and 5-year OS rates for IFI group were 67.6％,24.9％,and 15.0％,respectively,versus 73.7％,45.1％,and 26.0％ for ENI group (x2=5.04,P =0.025).The univariate stratified analysis showed that the LC and OS rates were significantly higher in the ENI group than in the IFI group for patients with middle-or lower-thoracic primary lesion or N0 disease (P=0.007,0.015;P=0.054,0.013).Conclusions For esophageal cancer patients with middle-or lower-thoracic primary lesion or without lymph node metastasis,prophylactic irradiation to the lymphatic drainage area can increase LC and OS rates.

Objective To compare the efficacy of elective nodal prophylactic irradiation ( ENI) and involved?field irradiation ( IFI) in radical radiotherapy for early?stage esophageal cancer and to determine the appropriate irradiation range for early?stage esophageal cancer. Methods The clinical data of 121 patients with early?stage esophageal cancer receiving radical radiotherapy in our hospital from January 2006 to December 2011 were collected and respectively analyzed. Sixty?one patients received ENI, and the other 60 patients received IFI. The Kaplan?Meier method was used to calculate local control ( LC) and overall survival ( OS) rates;the log?rank test was used for survival difference analysis and univariate prognostic analysis;the Cox regression model was used for multivariate prognostic analysis. Results The 1?, 3?, and 5?year LC rates in ENI group and IFI group were 81. 1%, 60. 1%, and 57. 5% vs. 64. 5%, 43. 9%, and 27. 2%, respectively ( P=0. 003 ) . The 1?, 3?, and 5?year OS rates in ENI group and IFI group were 86. 9%, 56. 8%, and 34. 8% vs. 86. 7%, 34. 3%, and 19. 1%, respectively ( P=0. 019) . The 1?, 3?,and 5?year overall failure rates in ENI group and IFI group were 22. 3%, 53. 8%, and 63. 2% vs. 43. 3%, 65. 8%, and 78. 8%, respectively ( P=0. 023) . Multivariate analysis showed that irradiation range was the influencing factor for LC and OS. Conclusions As for the radical radiotherapy for early?stage esophageal cancer, ENI can significantly increase LC and reduce locoregional failure, and therefore improve long?term OS.

Objective To analyze the efficacy of chemoradiotherapy in the treatment of esophageal carcinoma and its influencing factors,and to provide an optimal combination mode of chemoradiotherapy for treating esophageal carcinoma. Methods A retrospective analysis was performed on clinical data from 232 patients with esophageal carcinoma who were admitted to our hospital from January 2006 to December 2012 and received radical chemoradiotherapy. All patients received three?dimensional conformal radiotherapy or intensity?modulated radiotherapy as well as platinum?based chemotherapy. The overall survival ( OS ) and local control ( LC) rates were calculated using the Kaplan?Meier method and analyzed using the Logrank test. Univariate and multivariate prognostic analyses were made by the log?rank test and the Cox proportional hazard model,respectively. Results In all patients,the 1?,3?,and 5?year LC rates were 66?1%,42?2%, and 38?5%,respectively;the median LC time was 24?4 months;the 1?,3?,and 5?year OS rates were 73?3%, 37?2%,and 19?5%,respectively;the median OS time was 21 months. The univariate analysis revealed that T stage,N stage,clinical stage,irradiation range,and no less than 3 cycles of chemotherapy were influencing factors for OS ( P=0?000,0?000,0?000,0?030,0?001) and LC ( P=0?112,0?031,0?009,0?074,0?218) . The multivariate analysis revealed that N stage,clinical stage,and no less than 3 cycles of chemotherapy were independent prognostic factors for OS ( P=0?006,0?000,0?001) . Conclusions The LC and long?term OS rates in patients with early?stage esophageal carcinoma can be substantially improved by radical chemoradiotherapy. The irradiation range and no less than 3 cycles of chemotherapy improve the long?term survival in patients.

Background and purpose:p53 binding protein (53BP1) expresses in many normal and tumor cells. In vitroexperiments have conifrmed that inhibition of the protein expression of 53BP1 can effectively eliminate cycle arrest of tumor cells, and increase the radiosensitivity after irradiation. However, thein vivo experiment has not been re-ported. This study aimed to explore the effect of silencing 53BP1 gene on the growth and radiosensitivity to esophageal cancer cell ECA109 xenograft in nude mice.Methods:Forty-eight male BALB/c/nu nude mice were randomly divided into 6 groups: ECA109, ECA109/R, ECA109/N, ECA109/NR, ECA109/B and ECA109/BR. Three kinds of prepared cells (ECA109, ECA109/N and ECA109/B) were subcutaneously injected into the paw pads of mice (2×106/100 μL per mouse). The nude mice in ECA109/R, ECA109/NR, and ECA109/BR groups were irradiated with 15 Gy. Tumor growth was monitored every other day on the 6th day after injection. Tumor volume was measured with calipers. Theexpression levels of CHK1, CHK2 and phosphorylated CHK2-T68 protein were examined in different groups by West-ern blot. Apoptotic cell and cell cycle distribution were detected by lfow cytometry assay.Results:Visible tumors were detectable by day 7 after implantation, and the tumor volumes showed no signiifcant differences among all the groups (F=0.67, P=0.69). After irradiation with 15 Gy, tumor volumes in ECA109/BR group were smaller than those in other groups (P=0.03); the growth inhibition rate increased, but the relative growth rate decreased signiifcantly (P=0.01). The q value which relfected the radiosensitizing effect in ECA109/BR group was 1.45. The expressions of CHK1 and CHK2 at protein level in ECA109/BR group were not inlfuenced (P=0.71). However, the level of phosphorylated CHK2-T68 expression decreased signiifcantly after irradiation with 15 Gy (P=0.03). Cell cycle distribution and apoptosis were not signiifcantly different among all the groups (P=0.45).Conclusion:Silencing 53BP1 gene expression could inhibit the growth of esophageal cancer cell xenograft and increase the radiosensitivity to tumors in the nude mice.

Objective To explore the value of elective nodal prophylactic irradiation with intensity modulated radiotherapy(IMRT) for esophageal carcinoma.Screening patients who are suitable for elective nodal prophylactic irradiation (ENI),in order to improve locoregional control and overall survival.Methods The concurrent control study was conducted to esophageal cancer patients who were treated by definitive radiotherapy.A total of 148 patients finished treatment were identified.Seventy-four patients received ENI,while the other seventy-four patients received involved-field irradiation (IFI).Kaplan-Meier method was used for calculation of locoregional control rates and overall survival rates.The univariate and multivariate analysis of prognostic factors were also tested.Results The 1-,3-,and 5-year locoregional control rates of ENI group and IFI group were 72.5％,52.8％,50.6％ and 58.4％,35.8％,21.9％ (x2 =7.881,P ＜0.05),respectively.The 1,3,and 5 years survival rates of the ENI group and IFI group were 74.3％,44.2％,24.5％ and 68.9％,27.6％,15.9％ (x2 =1.903,P ＜ 0.05),respectively.In Cox multivariate analysis,clinical T stage,tumor location,different radiotherapy region were independent factors for the locoregional control of all patients,and clinical T,N stage,the length of esophageal barium meal and chemotherapy were independent factors for the overall survival of all patients.Conclusions Esophageal carcinoma patients treated with ENI could achieve better locoregional control than those treated with IFI.Esophageal carcinoma patients with early stage or middle thoracic lesion could benefit from ENI for local control and overall survival.