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Objective:To investigate the effect of hypoxia on the expression of forkhead box P3 (FOXP3) in human oral squamous cell carcinoma (OSCC) cells,and to clarify its possible epigenetic mechanism.Methods:Two kinds of OSCC cell lines,FaDu and OECM-1,were cultured under normoxic or hypoxic conditions for 18 h.The relative expression levels of FOXP3 mRNA and protein in the cells were detected by Real-time RT-PCR and Western blotting method.The histone modification levels on the FOXP3 gene promoter,including acetylation of histone 3 lysine 4 (H3K4ac),trimethylation of histone 3 lysine 4 (H3K4me3) and lysine 27 (H3K27me3),were analyzed by Chromatin Immunocipitation (ChIP) and quantitative PCR (ChIP-qPCR).The relative expression levels of histone deacetylase 3 (HDAC3) mRNA and inhibitory rates of FOXP3 mRNA expression in the HDAC3-knockdown FaDu cells were investigated by Real-time qPCR and ChIP-qPCR.Results:Compared with normoxic condition,the relative expression levels of FOXP3 mRNA in FaDu and OECM-1 cells under hypoxic condition were decreased by 65.6% and 75.7% (P<0.01).The Western blotting results indicated that compared with normoxic condition,the expression levels of FOXP3 protein in FaDu and OECM-1 cells under hypoxic condition were decreased.The ChIP experiment results showed that compared with normoxic condition,the levels of H3K4ac and H3K4me3 on FOXP3 gene promoter in FaDu cells were decreased under hypoxic condition (P<0.01),while the H3K27me3 level was not changed.In HDAC3-knockdown FaDu cells,compared with control cells,the inhibitory rates of the expressions of H3K4ac and H3K4me3 on FOXP3 gene promoter under hypoxia condition were decreased (P<0.05),so did expressions the FOXP3 mRNA expression (P<0.05).Conclusion:Hypoxia could suppress the expression of FOXP3 by HDAC3-mediated down-regulation of H3K4ac on FOXP3 gene promoter in the human OSCC cells.
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Objective To evaluate the inter-fraction setup error during the treatment with megavoltage computed tomography (MVCT) and provide theoretical basis for clinical target volume-planning target volume (CTV-PTV) margins for nasopharyngeal carcinoma (NPC) patients treated with tomotherapy.Methods Thirty-seven consecutive NPC patients treated with tomotherapy were prospectively enrolled for the study between February 2015 and September 2015.For each patient,one MVCT scan was obtained after conventional positioning,online correction and tomotherapy delivery daily,and the scan was registered to the planning CT to determine inter-fraction setup error.The expanding margin for PTV (MPTV) was calculated with the recipe:MPTV =2.5∑ + 0.76 (∑:systematic error;6:random error).Results The average absolute errors of the inter-fraction were (2.102 ± 0.040 6) mm,(1.490 ± 0.034 8) mm,(1.306 ± 0.335) mm and (1.392 ± 0.038 4) ° in the three dimensions.Gradual increases in both inter-fraction three-dimensional displacement were observed with time and treatment (P ＜ 0.05).The total MPTV ac counting for inter-error were 3.467 5 mm,2.979 5 mm and 2.888 5 mm.Conclusions Tomotherapy irradiation technology personalized MPTV should be adopted for the design of tomotherapy plan.Displacement increased as a function of time.
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Objective To assess the therapeutic efficacy of colonic tumor with targeted microbubbles encapsulated VEGFR2 monoantibody (mAb) combined with ultrasonic abrupted destroy.Methods Seventeen Balb/C nude mice with subcutaneous colonic carcinoma xenografts were divided into three groups:group A (5 mice) underwent contrast-enhanced ultrasonography (CEUS) examination and sham ultrasonic abrupted destroy;group B (6 mice) underwent lipid-microbubbles administration combined with ultrasonic irradiation;group C (6 mice) underwent VEGFR2 monoantibody-loaded microbubbles injection combined with ultrasonic irradiation.Red fluorescent protein(RFP) was labeled to all the nude mice model.Both CEUS and flurography were performed before and one week after abrupted destroy.The size,fluroscent area and fluroscent intensity(FI) and vessel density (VD) of each tumor were measured and compared.Results The parameters of length,fluroseent area,FI and VD of each tumor before abrupted destroy were no significant difference among three groups ( P ＞0.05).Parameters of post-sham ultrasonic abrupted destroy in group A were higher than those before sham ultrasonic abrupted destroy ( P ＜0.05).FI and VD in group B were significantly lower than those after abrupted destroy( P ＜0.05).There were no difference of length,fluroscent area of tumor in group B between pre- and post- ultrasonic abrupted destroy (P ＞0.05).Length,fluroscent area,FI and VD of each tumor in groups C were decreased significantly compared with post ultrasonic abrupted destroy ( P ＜0.01 ).There were significant difference of length,fluroscent area,FI and VD of each tumor among groups after ultrasonic abrupted destroy( P ＜0.05).Conclusions VEGFR2 mAb-loaded lipid microbubble combined with ultrasonic abrupted destroy can improve the therapeutic efficacy of colonic tumor.
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Objective To evaluate eutopic endometrial blood supply and vessels in patients with endometriosis(EMs) with contrast enhanced ultrasound (CEUS),to explore a newmethod to estimate vascularization of eutopic endometrium in EMs patients.MethodsThirty patients with EMs were enrolled and 20 patients who were diagnosed as other gynecological diseases as control.Informed consent of each case was obtained. AllcasesunderwentconventionalultrasoundexaminationandCEUSexamination.Characteristics of CEUS in each patients were observed and analyzed using off-line Qontrast software.Parameters of CEUS including peak intensity(PEAK),time to peak(TTP),regional blood volume(RBV),regional blood flow(RBF) were obtained automatically.ResultsComparing with control group,the eutopic endometrium in EMs group presented higher enhancement.The parameters of CEUS,including PEAK,TTP,RBV,RBF were (41.18 ± 3.29) dB,(29.01 ± 4.46) s,(29.07 ± 4.59) ml,(48.61 ± 5.35) ml/min in EMs group and (13.36 ± 2.34)dB,(24.59 ± 2.29)s,(26.51 ±- 3.80)ml,(48.71 ± 3.80)ml/min in controls respectively,the value of PEAK in EMs group was higher than that of controls ( P =0.000).Conclusions CEUS can be regarded as a new method which can be used to assess vascularization of eutopic endometrium in endometriosis.
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Objective To evaluate the correlation between quantitative parameters of contrastenhanced ultrasound(CEUS) and microvessel density (MVD) of rabbit atherosclerotic plaques.Methods Twenty-six male New Zealand rabbits were damaged abdominal aortic by balloon expanded,fed with high fat 12 weeks later.Thirty two plaques were detected using conventional ultrasound examination.The maximum area,luminal area stenosis rate and the maximum thickness of each plaque were recorded.CEUS was performed on 32 plaques and enhancement images were offline analyzed quantitatively by ACQ software and the parameters such as arrival time(AT),time to peak(TTP),basic intensity(BI),and peak intensity(PI) were acquired.Enhanced intensity (EI) was calculated according to the formula:EI =PI- BI.Then the experimentalrabbitswerekilledforpathologicalexaminationandCD34monoclonalantibody immunohistochemical detection.MVD was counted under the microscope.The correlation between the parameters of CEUS and MVD was analyzed.Based on pathological findings,the plaques were divided into vulnerable plaques and stable plaques.ResultsEI and MVD of vulnerable plaques group were significantly higher than that of stable plaques group [EI:(26.36 ± 1.44) dB vs (23.90 ± 2.92)dB,t =- 3.243,P =0.001 ；MVD:(5.23 ± 1.16)/mm2 vs (2.47 ± 1.12)/mm2,Z =- 4.378,P ＜0.001].The maximum area,luminal area stenosis rate,the maximum thickness and PI,AT,TTP had no significant difference between two groups( P ＞0.05).The value of EI showed significant positive correlation with MVD ( r =0.676,P ＜0.001).ConclusionsEI can accurately evaluate the angiogenesis of rabbit atherosclerotic plaques,which can be regarded as a useful index to distinguish between vulnerable and stable plaques.
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Objective To evaluate the application of transrectal ultrasound elastography (TRE) in the detection of prostate cancer(PCa).Methods One hundred and eighteen patients with suspected PCa were enrolled in this study.Each patient underwent transrectal ultrasound (TRUS), TRE and sonography guided prostatic biopsy on the same day.The accuracy rate of PCa detection using TRE were compared with the pathology results.Results Patients with PCa were detected in 62 of the 118 patients,including 48 cases by TRE and 38 cases by TRUS.The sensitivity for TRE (73.6％) was significantly higher than that of TRUS (52.8％) in periurethral zone(P ＜0.05).The sensitivity and specificity for strain ratio(SR) in predicting PCa were 85.7％ and 67.7％ respectively(cut off value：3.6,area under the curve 0.8, P ＜0.01,95％CI [0.68 - 0.92]).Conclusions TRE can detect PCa in the periurethral zone with good accuracy and has potential to increase ultrasound-based PCa detection rate.