Objective To explore the mechanism of Glaucocalyxin A(GLA)in inhibiting ovalbumin(OVA)-induced airway remodeling in asthmatic mice through the topoisomerase Ⅱ α(TOP2A)/cyclin-dependent kinase 1(CDK1)signaling pathway.Methods Forty Balb/c mice were randomly divided into 5 groups:the control group,the model group,the low-dose GLA group,the high-dose GLA group and the Dexamethasone group,with 8 mice in each group.The effect of GLA on airway remodeling was examined by immunohistochemical staining,ELISA and other methods,and bioinformatics methods were used to predict new targets of GLA.The action targets of TOP2A were screened using the STRING database,and the interaction relationship between the two was verified by co-immunoprecipitation.In vitro,GLA and siRNA were used to interfere with interleukin-4(IL-4)-stimulated human airway epithelial cells BEAS-2B.The expressions of TOP2A,epidermal growth factor receptor(EGFR),Integrin β1,focal adhesion kinase(FAK),β-catenin,CDK1 and DRP1 were detected by Western Blot.Results GLA intervention could significantly reduce OVA-induced asthma airway remodeling,airway smooth muscle thickening,collagen deposition around the airway,the number of eosinophils in alveolar lavage fluid,the expression of pro-inflammatory cytokines such as IL-4,and the level of serum IgE.The new target of GLA screened out was TOP2A,which was highly expressed in the lung tissue of the asthma airway remodeling model.GLA intervention could down-regulate its expression.In vitro,intervention with GLA and si-TOP2A could significantly down-regulate the expressions of IL-4-induced TOP2A,EGFR,Integrin β1,FAK and β-catenin.Further studies have found that TOP2A had an interaction relationship with CDK1.si-TOP2A could downregulate the expression of CDK1,and knockdown of CDK1 could significantly down-regulate the expression of phosphorylated DRP1.Conclusion GLA may alleviate asthma airway remodeling by targeting the TOP2A/CDK1 signaling pathway,providing experimental evidence for the clinical diagnosis and treatment of asthma airway remodeling in asthma.

Objective To explore the mechanism of Glaucocalyxin A(GLA)in inhibiting ovalbumin(OVA)-induced airway remodeling in asthmatic mice through the topoisomerase Ⅱ α(TOP2A)/cyclin-dependent kinase 1(CDK1)signaling pathway.Methods Forty Balb/c mice were randomly divided into 5 groups:the control group,the model group,the low-dose GLA group,the high-dose GLA group and the Dexamethasone group,with 8 mice in each group.The effect of GLA on airway remodeling was examined by immunohistochemical staining,ELISA and other methods,and bioinformatics methods were used to predict new targets of GLA.The action targets of TOP2A were screened using the STRING database,and the interaction relationship between the two was verified by co-immunoprecipitation.In vitro,GLA and siRNA were used to interfere with interleukin-4(IL-4)-stimulated human airway epithelial cells BEAS-2B.The expressions of TOP2A,epidermal growth factor receptor(EGFR),Integrin β1,focal adhesion kinase(FAK),β-catenin,CDK1 and DRP1 were detected by Western Blot.Results GLA intervention could significantly reduce OVA-induced asthma airway remodeling,airway smooth muscle thickening,collagen deposition around the airway,the number of eosinophils in alveolar lavage fluid,the expression of pro-inflammatory cytokines such as IL-4,and the level of serum IgE.The new target of GLA screened out was TOP2A,which was highly expressed in the lung tissue of the asthma airway remodeling model.GLA intervention could down-regulate its expression.In vitro,intervention with GLA and si-TOP2A could significantly down-regulate the expressions of IL-4-induced TOP2A,EGFR,Integrin β1,FAK and β-catenin.Further studies have found that TOP2A had an interaction relationship with CDK1.si-TOP2A could downregulate the expression of CDK1,and knockdown of CDK1 could significantly down-regulate the expression of phosphorylated DRP1.Conclusion GLA may alleviate asthma airway remodeling by targeting the TOP2A/CDK1 signaling pathway,providing experimental evidence for the clinical diagnosis and treatment of asthma airway remodeling in asthma.

Objective To investigate the hematological characteristics of patients with light β- thalassae-mia and rapidly identify different mutational genotypes. Methods RBC、Hb、MCV、MCH、MCHC、RDW-CV and HbA2 were studied in the 646 patients,the differences between β0/βN and β +/βN mutations were also compared. Results Most of them were microcytic hypochromic anemia. The most common genotype were β654/βN(33%)、β41-42/βN(32.5%)、β17/βN(14.4%)、β - 28/βN(10%)respectively,β0/βN were relatively higher. The differences in RBC、MCV、MCH、RDW-CV and HbA2 were significant between β0/βN and β +/βN. Compared with β +/βN patients,the MCV and MCH of β0/βN were significantly reduced,RDW-CV and HbA2 were significantly higher. Conclusion Light β- thalassaemia with different genotypes has its own unique hematological features and can be quickly and ef-fectively identified. Clinical efficiency can be improved through hematological analysis.

Objective To compare volumetric size, conformity index (CI), degree of inclusion (DI) of internal gross target volumes (IGTV) delineated on 4D-CT-MIP and PET-CT images for primary thoracic esophageal cancer. Methods Fifteen patients with thoracic esophageal cancer sequentially underwent enhanced 3D-CT, 4D-CT and PET-CT simulation scans. IGTVMIP was obtained by contouring on 4D-CT maximum intensity projection ( MIP). The PET contours were determined with nine different threshold methods (SUV≥2?0, 2?5, 3?0, 3?5), the percentages of the SUVmax(≥20%, 25%, 30%, 35%, 40%) and manual contours. The differences in size, conformity index (CI), degree of inclusion ( DI) of different volumes were compared. Results The volume ratios ( VRs) of IGTVPET2. 5 to IGTVMIP , IGTVPET20% to IGTVMIP, IGTVPETMAN to IGTVMIP were 0?86, 0?88, 1?06, respectively, which approached closest to 1. The CIs of IGTVPET2?0,IGTVPET2.5,IGTVPET20%,IGTVPETMAN and IGTVMIP which were 0?55, 0?56, 0?56, 0?54,0?55, respectively, were significantly larger than other CIs of IGTVPET and IGTVMIP (Z= -3?408-2?215,P <0?05). There were no statistical significance in the DIs of IGTVMIP and IGTVPET2.5,IGTVMIP and IGTVPET20%, IGTVMIP and IGTVPETMAN(0?77,0?82,0?71,0?67, 0?68,0?82,P>0?05). Conclusions The targets delineated based on SUV threshold setting of≥2?5, 20% of the SUVmax and manual contours on PET images correspond better with the target delineated on maximum intensity projection of 4D-CT images than other SUV thresholding methods.

Objecttve To compare the positional and volumetric differences of planning target volumes(PTVs)based on axial three-dimensional CT(3D-CT)and four-dimensional CT(4D-CT)for the primary tumor of non-small cell lung cancer(NSCLC).Methods Sixteen NSCLC patients with lesions located in the upper lobe and 12 patients with lesions in middle and lower lobes,totally 28 patients, initially underwent three-dimensional CT scans followed by 4D-CT scans of the thorax under normal free breathing.PTVvector was defined on gross tumor volume (GTV) contoured on 3D-CT and its motion vector. The clinical target volumes(CTVs)were created by adding 7 mm to GTVs,then, internal target volume (ITVs)were produced by enlarging CTVs isotropically based on the individually measured amount of motion in the 4D-CT,lastly,PTVs were created by adding 3 mm setup margin to ITVs. PTV4D was defined on the fusion of CTVs on all phases of the 4D data.The CTV wag generated by adding7 mm to the GTV on each phase.then,PIVs were produced by fusing CTVs on 10 phases and adding 3 mm setup margin.The position of the target center,the volume of target and the degree of inclusion(DI)were compared reciprocally between the PTVvector and the PTV 4D The difference of the position,volume and degree of inclusion of the targets between PTVvecter and PTV4D were compared,and the relevance between the relative characters of the targets and the three-dimensional vector was analyzed based on the groups of the patients. Results The median of the 3 D motion vector for the lesions in the upper lobe was 2.8 mm, significantly lower than that for the lesions in the middle and lower lobe ( 7.0 mm, z = - 3. 485, P < 0. 05 ). In the upper lobe group there was only significant spatial difference between the PTVvector and PTV4D targets in the center coordinate at the x axe (z = -2. 010, P < 0. 05 ), while in the middle and lower lobes there was only significant spatial difference between the PTVvector and PTV4D targets in the center coordinates at the z axe (z = -2. 136,P <0.05). The median of ratio of PTV4D and PTVvector, of the upper lobe group was 0. 75, significantly higher than that of the middle and lower lobes group (0. 52, z = - 2. 949, P < 0. 05 ).A significant correlation was found for the motion vector and the ratio of PTV and PTV4D in both groups ( r = - 0. 638, - 0. 850, P < 0. 05 ). For all patients, the median of D[ of PTV4D in PTVvector was 66. 39% ,while the median of DI of PTVvector, in PTV4D was 99. 55% , both showed a positive significant correlation with the motion vector (r = -0. 814,0. 613 ,P < 0. 05). Conclusions PTV4D defined based on 4D-CT simulation images is obviously less than PTV defined based on 3D-CT simulation images. The ratio and DI of both targets are related with the three-dimensional motion vector of the tumor.

Objective To compare positional and volumetric differences of internal gross tumor volume (IGTV) delineated separately by three approaches based on four-dimensional CT (4DCT) for the primary tumor of non-small cell lung cancer (NLCLC). Methods Twenty-one patients with NLCLC underwent big bore 4DCT simulation scan of the thorax. IGTVs of the primary tumor of NSCLC were tumor on the MIP images were delineated to produce IGTVMIP. The position of the target center, the volume of target, the degree of inclusion (DI) and the matching index (MI) were compared reciprocally between IGTV10, IGTVEI+EE and IGTVMIP. Results Average differences between the position of the center of IGTVs on direction of x,y and z axes were less than 1 mm, with no statistically significant difference. The volume of IGTV10 was larger than that of IGTVEI+EE, the difference was statistically significant (t=2.37,P=0.028);the volume of IGTV10 was larger than that of IGTVMIP, but the difference was not statistically significant(t=1.95 ,P=0.065). The ratio of IGTVEI+EE with IGTV10, IGTVMIP with IGTV10 were 0.85±0.08 and 0.92±0.11, respectively. DI of IGTVEI+EE in IGTV10, IGTVMIP in IGTV10 were 84.78% ± 8. 95% and 88.47% ±9.04%. MI between IGTV10 and IGTVEI+EE, IGTV10 and IGTVMIP were 0.85 ±0.09, 0.86±0. 09, respectively. Conclusions The center displacement of the IGTVs delineated separately by the three different techniques based on 4DCT images are not obvious; IGTVEI+EE and IGTVMIP can not replace IGTV10 , however , IGTVMIP is more close to IGTV10 comparing to IGTVEI+EE . The ratio of GTVEI+EE with IGTV10 is correlated to the tumor motion vector. As the vector increases, the ratio of GTVEI+EE with IGTV10decreases, especially for small tumors.

Objective To investigate the correlations of the whole breast displacement in different respiratory cycle during free breathing (FB) following breast-conserving surgery to the displacement of selected skin marker,nipple,and selected surgical clip based on four-dimensional computed tomography (4D-CT).Methods Thirteen breast cancer patients who had undergone breast-conserving surgery received whole breast intensity-modulated radiotherapy (IMRT).Respiration-synchronized 4D-CT image data were gathered during FB and were exported to the Varian Eclipse treatment planning system,and the whole breast target,nipple,superior clip,and metal marker on the skin at the anterior body midline were delineated on the CT images of ten phases of the respiratory cycle by the same radiotherapist based on the same delineating criteria.The displacement distances of the delineated target in the mediolateral (x),anteroposterior (y),and superoinferior (z) axles were achieved,and the correlations of the whole breast target displacement to the displacement of the clip,nipple,and skin marker were analyzed.The ipsilateral lung was delineated on the CT images of every phase of the respiratory cycle,and the changes in ipsilateral lung volume were analyzed during the respiratory cycle relative to the displacement of the breast.Results The maximal displacement distances of the whole breast target in the x,y,and z axles during FB were 0.71,0.76 and 1.29 mm,respectively ( F =5.755,P ＜ 0.05 ).There was no relationship between the three-dimensional (3D) displacement of the whole breast and the volume of the whole breast (r =-0.264,P ＜ 0.05 ),and there was no relationship between the displacemeat of the whole breast and the volume change of the ipsilateral lung ( r =0.346,P ＜ 0.05).There was no significant difference among the mean target displacement distances in 3 axles,and among 2 selected successive end-inspiration (EI) phases and 3 selected successive end-expiration (EE) phases.There was no significant difference between the volumes of the whole breast targets at the selected El and EE phases.There was no relationship between the displacement of the whole breast target and the displacement of the nipple,skin marker or superior clip in the cavity along the x- and z-axles.Along the y-axle,8/13,7/11 and 9/13 of the patients showed displacement of the whole breast target relative to the displacement of the nipple,skin marker and superior clip respectively.However,according to a population-based analysis,the displacement of the whole breast target was only significantly associated with the displacement of the superior clip ( r =0.657,P ＜ 0.05 ).Conclusions The clip registration is more credible and sensitive than a skin marker or the nipple for measuring and correcting the displacement of the whole breast target during radiotherapy.