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Objective:To investigate the effect of decreasing DKC1 gene expression on radiosensitivity of HeLa cells.Methods:A cell model with low expression of DKC1 gene was established by shRNA technology with lentivirus as vector, and the interference efficiency was verified by RT-PCR and Western blot assay. Cells were divided into two groups of interference (Lv-shDKC1) and its negative control. Telomerase activity was detected by TRAP-ELISA, and telomere length was measured by Real-time PCR. Cell survival was obtained through clone formation assay and fitted by multi-target single-hit model, and radiobiological parameters ( D0, Dq, N, SF2) and radiosensitization ratio (SER) were calculated. Results:After DKC1 interfering, the expression levels of mRNA and protein of DKC1 in HeLa cells were significantly decreased by (71.330±4.112)% ( t=25.53, P<0.05) and (35.520±3.804)% ( t=4.833, P<0.05), respectively. Compared with the blank control group and negative control group, the telomerase activity of Lv-shDKC1 group decreased significantly from 0.900±0.044 and 0.897±0.031 to 0.713±0.021 ( F=31.44, P<0.05), the relative telomere length was significantly decreased from 4.233±0.306 and 4.633±0.379 to 2.667±0.404 ( F=39.15, P<0.05). The telomerase activity and relative telomere length of blank control group and Lv-shDKC1 negative control group had no significant difference( P>0.05). SF2 in the interference group (0.571±0.006) was significantly lower than that of the blank control group (0.861±0.009) and the Lv-shDKC1 negative control group (0.807±0.002) ( F=1812, P<0.05), and the radiosensitization ratio (SER) of shDKC1 interference was 1.508. Conclusions:Downregulation of DKC1 in human cervical cancer HeLa cells enhances the radiosensitivity through inhibiting the activity of telomerase and shortening the length of telomere. DKC1 gene may become a new target of radiosensitization.
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Objective To observe the correlation between CD4+ CD29+ T cells and metastasis and radiotherapy for patients with pulmonary adenocarcinoma. Method Seventy-one patients with lung adenocarcinoma, 93 patients with lung adenocarcinoma ,76 cases of chronic obstructive pulmonary disease (COPD),63 cases of healthy volunteers were enrolled. Frequencies of blood CD4+ CD29+ T cells and their intracellular necrosis factor alpha(TNF-α)and interleukin 1(IL-1)were compared. Compare TNF-α,IL-1,integrin beta 1 and vascular endothelial growth factor(VEGF)levels in the patients with transferred pulmonary adenocarcinoma or with non-transferred pulmonary adenocarcinoma and their changes with the treatment of radiotherapy. Results the patients with lung adenocarcinoma and non lung adenocarcinoma were significantly higher than that of COPD and health group,and patients with lung adenocarcinoma is significantly higher than patients with non lung adenocarcinoma (P<0.05);Integrin beta 1,VEGF and CD4+CD29+T cells,TNF-αand IL-1 level in patients with lung adeno-carcinoma metastasis were significantly higher than non-transferred group(P < 0.05);After radiotherapy,CD4+CD29+T cells,TNF-αand IL-1 in patients with lung adenocarcinoma were significantly lower than before(P<0.05);CD4+ CD29+ T cells,TNF alpha and IL-1 with integrin beta 1 and VEGF had significantly positive correlations. Conclusion CD4+CD29+T cells and cytokines increase significantly in the blood of patients with lung adenocarci-noma,and are related to the prognosis of metastasis and radiation therapy,which has important clinical significance.
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Objective To evaluate the efficacy and toxicity of induction chemotherapy with nedaplatin and docetaxel plus concurrent intensity?modulated radiotherapy ( IMRT) with nedaplatin or cisplatin in the treatment of locally advanced nasopharyngeal carcinoma (NPC). Methods A total of 223 patients with pathologically diagnosed locally advanced NPC in five treatment centers from 2011 to 2012 were randomly divided into two groups. In the test group, one hundred and thirteen patients received two cycles of induction chemotherapy with docetaxel (65 mg/ m2 on day 1) and nedaplatin (80 mg/ m2 on day 1) plus concurrent IMRT with nedaplatin (40 mg/ m2 on day 1). In the control group, 110 patients received two cycles of induction chemotherapy with the same regimens plus concurrent IMRT with cisplatin (40 mg/ m2 on day 1). The survival rates were calculated with the Kaplan?Meier method and the differences in the survival rates between the two groups were analyzed using the log?rank test. Comparison of the incidence rates of adverse reactions between the two groups was made by the chi?square test. Results The follow?up rate was 99?? 1%.The response rates at 3 months after treatment in the two groups were both 100%. The 2?year local recurrence?free, regional recurrence?free, distant metastasis?free, and overall survival rates were 94?? 0%, 94?? 2%, 88?? 2%, and 90?? 3%, respectively, in the test group, versus 93?? 4%, 94?? 1%, 86?? 7%, and 87?? 3% in the control group ( P= 0?? 757、 0?? 478、 0?? 509、 0?? 413). The incidence rates and severity of leucopenia, neutropenia, and thrombocytopenia were significantly higher in the test group than in the control group ( P= 0?? 027 , 0?? 028 , 0?? 035 ) . The incidence rates and severity of hemoglobin reduction and nausea /vomiting were significantly lower in the test group than in the control group (P= 0?? 000,0?? 023). There were no significant differences in the incidence rates of mucositis and xerostomia between the two groups ( P=0?? 483,0?? 781). Conclusions The short?term efficacy of induction chemotherapy with nedaplatin and docetaxel plus concurrent IMRT with nedaplatin is similar to that with cisplatin in the treatment of locally advanced NPC. The mild gastrointestinal reactions can be tolerated by patients. However, the severe myelosuppression should be closely monitored during the treatment.
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Objective To explore the effect and underlying mechanism of Sarcandra glabra Nakai on radiation pneumonnopathy in miniature pigs irradiated with 60Co γ-rays.Methods Totally 60 miniature pigs were randomly divided into blank control group,irradiated group with 15 Gy of 60Co γ-rays,and intervention group administered with Sarcandra glabra Nakai at the dosage of 0.3 g/kg before 15 Gy irradiation.At 4,8,12 and 24 weeks post-irradiation,the respiratory rate and weight were observed,and a portion of the right lung tissue was taken out to conduct HE and Masson staining examination,while the expressions of TGF-β1 and TNF-α as well as the content of hydroxyproline (HP) were detected.Results The respiratory rate,the lung coefficient and the content of HP in the irradiated group showed an increasing trend at 4,8,12 and 24 weeks post-irradiation and significantly higher than those in other two groups (F =21.035,146.014,32.610,P ＜ 0.05).Those indicators were indistinguishable between the control group and intervention group at 4 and 8 weeks post-irradiation (F =0.055,2.456,5.581,P ＞ 0.05),while the indicators of the irradiation group were significantly higher than that of intervention group (F =91.897,93.149,83.487,P ＜0.05) at 12 and 24 weeks post-irradiation.The pulmonary histopathological results showed that no inflammatory response or fibrosis was found in the control group and intervention TGF-β1 and TNF-α in the intervention group were negatively expressed at 4 and 8 weeks post-irradiation and had low expressions at 12 and 24 weeks post-irradiation which was lower than that of the irradiated group.Conclusions Sarcandrae has protective effect against radiation-induced lung injury in miniature pigs probably by inhibiting the expressions of TNF-α and HP.
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Objective To investigate the relationship between RKIP expression and the efficiency of radiotherapy in NPC patients and evaluate the possibility of using RKIP as a predictor of radiosensitivity.Methods A total of 180 patients with NPC in Sun Yat-sen University Cancer Center without evidence of distant metastasis at initial diagnosis were enrolled in this study,who had received intensity-modulated radiotherapy alone.Patients were classified into 2 groups according to criteria below:patients with biopsy proven recurrent diseases occurring at nasopharynx and/or neck within 5 years after radiotherapy were classified as the radioresistant group.The pathological type at relapse was the same as the previous one before treatment.Patients with a minimum follow-up of 5 years after radiotherapy without evidence of recurrence at the original site of the tumor were classified as the radiosensitive group.Patients in the 2 groups were matched according to the factors related with radiosensitivity.RKIP was examined by immunohistochemical staining before radiotherapy.The relationship between RKIP expression and the effect of radiotherapy were analyzed.Results The positive rate of the RKIP expression in the radiosensitive group versus the radioresistant group was 80.0％ versus 26.7％.The positive rate (x2 =12.498,P ＜0.01) and the intensity of the RKIP expression (x2 =51.429,P ＜ 0.01) were significantly different between 2 groups with a negative correlation with radio-resistance to NPC (r =-0.344,-0.535,respectively,P ＜ 0.01).Based on the RKIP expression,the radiosensitivity,specificity,accuracy,positive predictive value,negative predictive value,false positive and false negative were predicted as follows:80.0％,73.3％,77.2％,75.0％,78.6％,26.7％,and 20.0％,respectively.Conclusions RKIP protein shows negative correlation with radioresistance to NPC and could serve as a biomarker in preliminarily screening the intrinsic radiosensitivity of NPC.
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Objective To evaluate the treatment efficacy,toxicities and prognostic factors of nasopharyngeal carcinoma ( NPC ) treated with intensity modulated radiation oncology ( IMRT ).Methods Between January 2006 and August 2008,300 patients with pathologically diagnosed NPC from 6 center received IMRT.The number of patients with stage Ⅰ,Ⅱ,Ⅲ and Ⅳa+b disease (UICC/AJCC 2002 staging system) were 6,45,141 and 108,respectively.The prescription doses were as follows:70-74 Gy/30f toplanning target volume of primary nasopharynx tumor ( PTVRλ),68-70 Gy/30f to planning target volume of positive lymphnode (ptvnd),60-64 Gy/30f to higher risk region (PTV1),50-54 Gy/30f to lower risk region (PTV2).Patients with stage Ⅲ and Ⅳa+b disease also received cisplatin-based chemotherapy.Cox method was used for Multivariate analysis.ResultsThe follow-up rate was 99.7％.The 4-year rate of local control,regional control,metastasis-free survival (DMFS),disease-free survival (DFS) and overall survival (OS) was 94.0％,95.5％,87.4％,80.8％,86.1％,respectively.Mucositis was the most severe acute toxicity,with 18.0％grade 1,48.7％grade 2,33.3％grade 3.No patient suffered from grade 4mucositis.Xerostomia was the most common late toxicity,with 12.0％ grade 0,75.7％ grade 1,12.3％grade 2.No grade 3-4 xerostomia was observed.There were 18,15 and 42 patients failed in local,regional and distant metastasis,respectively.Multivariate analysis showed that N stage was the only prognostic factor for OS (x2 =5.17,P=0.023),DMFS (x2 =6.91,P=0.009) and DFS (x2 =8.15,P=0.004) in these patients.ConclusionsIMRT can improve the treatment efficacy of NPC.The acute and late toxicities were tolerated.Distant metastasis becomes the main treatment failure.N stage is a significant prognostic factors.
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Objective To discuss the feasibility of Monte Carlo N-particle transport code(MCNP)simulated calculation.Methods The calculation in water phantom was contrasted with the practical measurements and the reported values using the percent depth dose(PDD)curve and normal peak scatter factor.Results There Was no significant difference between calculated and measured results in the 10 cm×10 cm field(t＝-0.41,P>0.05),however,there were significant differences in the 5 cm×5 cm field(t=7.2,P<0.05)and in the 12 cm×12 cm field(t=-4.6,P<0.05).There was no significant difierence between the calculated results and the reported values(t=-1.91,P>0.05).In the same radiation field,the PDD decreased as the depth increased,but increased as the size of the radiation field increased at the same depth.PDD and normal peak scatter factor were both important parameters for calculation of prescribed dose.Conclusions It is possible to establish a set of accurate and comprehensive percent depth doses and normal peak scatter factor parameters so as to provide the basis of clinical use, quality assurance and quality control for radiotherapy.
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Objective To study more reasonable method of radiation barrier for cervical cancer treated with external radiation. Methods The distance from cervical canals to tumor's margin in x axis was measured by B ultrasonic, around uterus were evaluated by physical examination.and the dose around uterus for patient who used fixed lead brick for radiation barrier when treated with external radiation were calculationed by TPS, in 39 case of cervical cancer treated with external radiation combine with intracavitary irradiation. Pay attention to the lower dose area around uterus. Results It might bring about lower dose area around uterus who used fixed lead brick for radiation barrier when treated with external radiation,and mass might be in above area. Conclusion Cervical cancer treated with external radiation with source axial distance (SAD), and radiation barrier with lead brick individuate may help for to avoid the lower dose area around uterus.
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Objective:To study the expression and significance of transforming growth factor-β1 (TGF-β1) in nasal polyps.Method:Expression of TGF-β1 in nasal polyps from 34 patients and middle turbinates from 30 patients with deviation of nasal septum were prospectively studied with immunohistochemistry.Each tissue section was observed under optical microscope.Result:①The TGF-β1 positivity in extracellular matrix and in cells in the stroma was significantly higher in nasal polyps than in middle turbinates (P＜0.01).②The distribution and shape of TGF-β1 expressing cells in nasal polyps was similar to that of eosinophil,their positivities were significantly correlative (P＜0.05).③The positivity of TGF-β1 did not correlate with clinical type of nasal polyps (P＞0.05),eosinophil infiltration correlated significantly with clinical type of nasal polyps(P＜0.05).Conclusion:①The TGF-β1 may contribute to some of the pathologic changes observed in nasal polyps,such as thickening of the epithelial basement membrane and stromal fibrosis.②Eosinophils in nasal polyps represent a major source of TGF-β1.③Eosinophils infiltration may play a prominent role in the development and recurrence of nasal polyps.
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AIM: To investigate the effect of 17?-estradiol on insulin action in cultured C2C12 mytoblasts. METHODS: C2C12 mytoblasts were cultured in 35 mm wells of six-well culture plate in an atmosphere of 5% CO 2 at 37℃ in DMEM supplemented with 10% FBS and penicillin/streptomycin(1?10 5 U/L) to reach 80% confluence. Insulin-resistance C2C12 mytoblasts were obtained by incubating the cells for 24 hours in the presence of a high concentration (5?10 -7 mol/L) of insulin. After treatmented with 17?-estradiol (1 nmol/L and 10 nmol/L, respectively) for 24 hours, C2C12 mytoblasts were performed to measure insulin-stimulated 2-DG uptake and GS, PFK, PK activities. RESULTS: 17?-estradiol enhanced the capacity of insulin-stimulited 2-DG uptake, increased the GS, PFK and PK activities and prevented insulin-induced resistance in cultured C2C12 mytoblasts. CONCLUSION: 17?-estradiol potentiates insulin action and preventes insulin-induced resistance in cultured C2C12 mytoblasts.
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AIM:To investigate the influence of ovariectomy and estrogen replacemeot treatment on profile of gene expression in myocardium by cDNA microarray,and to characterize the targeting genes of estrogen.METHODS:cDNA microarray containing 1 400 rat cDNAs was used to study the genes differentially expressed in myocardium between sham(Ⅰ),ovariectomy(Ⅱ,OVX)and estrogen replacement treatment(Ⅲ,OVX+E2)group.Then down-regulated genes in myocardium of OVX rats were further confirmed by RT-PCR.RESULTS:177 genes were differentially expressed in myocardium between sham and OVX rats,with 91 genes up-regulated and 86 genes down-regulated in OVX rats.164 genes were differentially expressed in myocardium between OVX and OVX+E2 rats,with 113 genes up-regulated and 54 genes down-regulated in OVX rats.There were 54 genes differentially expressed in OVX compared to sham and OVX+E2.They are involved in membrane channels and transporters(18),cell receptors(9),intracellular transducers/effectors/modulator(7)and metabolism(6).Most of the genes(45)were down-regulated in OVX rats and up-regulated in OVX+E2 rats.RT-PCR test confirmed the results of cDNA microarray.CONCLUSIONS:Long-term estrogen replacement may influence the expression of genes involved in membrane channels and transporters,cell receptors,intracellular transducers/effectors/modulator and metabolism.Long-term estrogen replacement has some beneficial effects on ionic concentration and cardiac function which partially comes from the results of influence of expression on Na+,K+-ATPase and Na+/H+ exchanger.Estrogen has an inhibitory effect on the expression of dopamine receptor,which partially clarify the myocardial protection of estrogen.
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AIM: To study the molecular mechanism in modulation of expression of insulin receptor substrate-1 and-2 (IRS-1,-2) by estrogen and high concentration of insulin. METHODS: The 5′-regulatory regions of IRS-1 and IRS-2 gene were cloned into the pGL3 plasmid with luciferase reporter, and the clones were transfected into HeLa cells. The cells were incubated with estradiol (1 nmol/L) and high concentration of insulin (100 nmol/L). The relatively transcriptional activity of the 5′-regulatory regions of IRS-1 and IRS-2 gene was detected. RESULTS: It was found that the relatively transcriptional activity of the 5′-modulatory regions of IRS-2 reduced markedly after cells were incubated with 100 nmol/L insulin (P