The mechanism study on Chaihudaxiong mixture in the treatment of coronavirus disease 2019 with network pharmacology approach / 药学实践杂志

Objective To investigate the pharmacological mechanism of Chaihudaxiong mixture in the treatment of coronavirus disease 2019 (COVID-19) based on a network pharmacology approach. Methods The effective ingredients and targets of Chaihudaxiong mixture were collected from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP). The targets’ names were standardized by Uniprot database. Genes associated with coronavirus were obtained from the GeneCards and OMIM, which were intersected with effective therapeutic targets. A "herbs-ingredients-targets" network was compiled and analyzed by Cytoscape 3.7.2. The protein-protein interaction of the targets was analyzed by String. The GO gene annotation and KEGG signaling pathway analysis were performed using related packages of the R software. Results A total of 165 active ingredients and 51 targets were collected. Further analysis revealed that the main active ingredients were β-sitosterol and 11 flavonoids. The core targets were CASP3, MAPK3, IL-6, MAPK8, IL-10, CXCL8, MAPK1 and IL-1B. A total of 1722 GO entries were obtained from the GO gene annotation (P<0.05), including 1612 entries for biological processes, 30 entries for cell composition, and 80 entries for molecular functions. 156 signaling pathways (P<0.05) were obtained with KEGG signaling pathway screen. The important signaling pathways were AGE-RAGE signaling pathway in diabetic complication, Influenza A, IL-17 signaling pathway, TNF signaling pathway and hepatitis B. Conclusion This study revealed the synergistic features of multi-component, multi-target, and multi-pathway of Chaihudaxiong mixture in the treatment of COVID-19, which provided an important scientific basis for further understanding the mechanism of Chaihudaxiong mixture in the treatment of COVID-19.

Differentially expressed microRNA and target gene analysis in patients with postmenopausal osteoporosis / 中华创伤杂志

Objective To analyze the differentially expressed microRNA (miRNA) and their target genes in peripheral blood and bone tissue of postmenopausal osteoporosis (PMOP),and provide basis for the study of pathogenesis as well as biomarkers identification of PMOP.Methods Two miRNA datasets of PMOP from the public platform NCBI-GEO DataSets were obtained,including GSE64433 (the miRNA expression profile of peripheral blood samples,including 23 PMOP patients and 25 controls) and GSE74209 (the miRNA expression profile of the femoral neck bone tissue sample,including six PMOP patients and six controls).R/Bioconductor was performed for data analysis and differentially expressed miRNA screening,and miRNAs with fold change ＞ 2 & P ＜ 0.05 between osteoporosis and controls were selected as differentially expressed miRNA.The miRNA target gene prediction was performed by combining TargetScan,miRDB and miRTarBase databases.The miRNA-target gene regulatory network was constructed and analyzed by Cytoscape.Results There were 224 differentially expressed miRNAs (75 up-regulated miRNAs and 149 down-regulated miRNAs) in the peripheral blood samples of PMOP group (GSE64433 dataset) and 132 differentially expressed miRNAs (58 up-regulated miRNAs and 74 down-regulated miRNAs) in the femoral neck bone tissue of PMOP group (GSE74209 dataset).We combined the results from the two datasets and obtained 8 miRNAs down-regulated in both datasets,and the 8 miRNAs regulated a total of 327 target genes,and 10 of these target genes were co-regulated by two miRNAs.Conclusions The core miRNAs and the target genes regulated by multiple miRNAs in the regulatory network may play important roles in the pathogenesis of PMOP and have potential application values as molecular biomarkers of disease.These findings are meaningful for the studies of PMOP pathogenesis,biomarkers screening and prevention of osteoporotic fractures.

Differentially expressed microRNA and target gene analysis in patients with postmenopausal osteoporosis / 中华创伤杂志

Objective@#To analyze the differentially expressed microRNA (miRNA) and their target genes in peripheral blood and bone tissue of postmenopausal osteoporosis (PMOP), and provide basis for the study of pathogenesis as well as biomarkers identification of PMOP.@*Methods@#Two miRNA datasets of PMOP from the public platform NCBI-GEO DataSets were obtained, including GSE64433 (the miRNA expression profile of peripheral blood samples, including 23 PMOP patients and 25 controls) and GSE74209 (the miRNA expression profile of the femoral neck bone tissue sample, including six PMOP patients and six controls). R/Bioconductor was performed for data analysis and differentially expressed miRNA screening, and miRNAs with fold change>2 & P<0.05 between osteoporosis and controls were selected as differentially expressed miRNA. The miRNA target gene prediction was performed by combining TargetScan, miRDB and miRTarBase databases. The miRNA-target gene regulatory network was constructed and analyzed by Cytoscape.@*Results@#There were 224 differentially expressed miRNAs (75 up-regulated miRNAs and 149 down-regulated miRNAs) in the peripheral blood samples of PMOP group (GSE64433 dataset) and 132 differentially expressed miRNAs (58 up-regulated miRNAs and 74 down-regulated miRNAs) in the femoral neck bone tissue of PMOP group (GSE74209 dataset). We combined the results from the two datasets and obtained 8 miRNAs down-regulated in both datasets, and the 8 miRNAs regulated a total of 327 target genes, and 10 of these target genes were co-regulated by two miRNAs.@*Conclusions@#The core miRNAs and the target genes regulated by multiple miRNAs in the regulatory network may play important roles in the pathogenesis of PMOP and have potential application values as molecular biomarkers of disease. These findings are meaningful for the studies of PMOP pathogenesis, biomarkers screening and prevention of osteoporotic fractures.

Diagnostic and prognostic roles of loss of CIC protein expression in oligodendroglial tumors / 中华病理学杂志

Objective@#To investigate the usefulness of loss of CIC expression as the prescreening detection of 1p/19q co-deletion in the diagnosis of oligodendroglial tumors and its prognostic implication.@*Methods@#The retrospective study included 113 oligodendroglial tumors diagnosed in the Department of Pathology, Xuanwu Hospital, Capital Medical University. Expression of CIC protein was detected by immunohistochemistry, and the 1p/19q co-deletion by fluorescence in situ hybridization in all the tumors; and the correlation of the loss of protein and 1p/19q co-deletion with prognosis was assessed.@*Results@#The rate of negative CIC protein expression was 59.3% (67/113) in 113 oligodendroglial tumors. CIC protein expression was differentially lost in various gliomas, 85.7% (42/49) in pure oligodendrogliomas and 39.1% (25/64) in mixed oligodendroglial tumors (P<0.01). The loss of CIC protein expression showed a sensitivity of 76.1% (54/71), specificity 71.1% (27/38), false positive rate of 16.9% (11/65), and a false negative rate of 38.6% (17/44). In 63 cases integrated diagnosis as oligodendroglial tumors with mutant IDH and 1p/19q co-deletion, the loss of CIC protein expression was 81.0% (51/63); the sensitivity and specificity were increased to 81.0% (51/63) and 76.9% (20/26), and the false positive rate and false negative rate decreased to 10.5% (6/57) and 37.5% (12/32), respectively. By using Kaplan-Meier analysis, the CIC negative group showed a trend towards better outcome than the CIC positive group, but there was no statistical difference (overall survival: P=0.218; progression free survival: P=0.249).@*Conclusions@#Detection of the lost CIC protein expression can predict the chromosome 1p/19q co-deletion. In oligodendroglial tumors with IDH mutant and 1p/19q co-deletion, there is no relation between prognosis and CIC protein expression.

Exercise effects on pain relief and extracellular matrix production in a rat model of intervertebral disc degeneration / 中国组织工程研究

BACKGROUND: Exercise has been proved to accelerate the proliferation of intervertebral disc cells and extracellular matrix production in healthy rats. For the degenerative intervertebral disc, whether exercise also has positive effects on its cell proliferation, extracellular matrix production or pain relief remains unclear. OBJECTIVE: To investigate the effect of exercise on the extracellular matrix production in a rat model of intervertebral disc degeneration.METHODS: A rat model of intervertebral disc degeneration was prepared by Freund's complete adjuvant injection into the intervertebral disc at L5-6 levels. Then, the model rats were allowed to have a rest for 2 weeks. All rats were then randomly divided into exercise and control groups. Rats in the exercise group were forced to run every day, while the controls allowed free activities in the cage. The behavioral tests were performed at 7, 14, 28, 42, 56 and 70 days after modeling; meanwhile, the intervertebral disc samples were collected used for alcian blue staining and immunohistochemical staining to detect the levels of proteoglycan, aggrecan and collagen type Ⅱ in the intervertebral disc cells, respectively.RESULTS AND CONCLUSION: Vocalization threshold on the rat back of punctured disc was significantly decreased, while grooming and wet-dog shaking were significantly increased at 7 days after modeling compared with the baseline (P < 0.05), suggesting that Freund's complete adjuvant injection successfully induces disc degeneration, hyperalgesia and abnormal behaviors. Further, the vocalization threshold and wet-dog shaking in the exercise group showed significant improvement compared with the control group after 14 days of exercise (P < 0.05), while the grooming was significantly reduced until the 28th day (P < 0.01), indicating that exercise can alleviate pain caused by disc degeneration in model rats. At 21 days after modeling, the levels of proteoglycan, aggrecan and collagen type Ⅱ in the nucleus pulposus and annulus fibrosus were significantly decreased compared with the baseline (P < 0.01), indicating the occurrence of disc degeneration. After 14 days of training, the levels of proteoglycan, aggrecan, and collagen type Ⅱ in the nucleus pulposus and annulus fibrosus in the exercise group were significantly increased compared with the control group (P < 0.01). Moreover, after 8-week exercise, the level of proteoglycan in the nucleus pulposus and annulus fibrosus in the exercise group was increased by 4-5 times compared with the control group, and levels of aggrecan and collagen type Ⅱ in the nucleus pulposus in the exercise group also was increased by 3-4 times compared with the control group. To conclude, exercise can promote extracellular matrix increased by production by increasing the levels of proteoglycan, aggrecan, and collagen type II in the degenerative intervertebral disc.

Piperine inhibits Ang Ⅱ-induced cellproliferation and migration of rat airway smooth muscle cells / 基础医学与临床

Objective To explore the effects of piperine on cell proliferation and migration in angiotensin Ⅱ (Ang Ⅱ)-treated rat airway smooth muscle cells (ASMCs).Methods The primary ASMCs of rats were cultured by improved tissue-piece digestion inoculation and trypsin digestion.MTT assay was used to detect the effects of Ang Ⅱ and Ang Ⅱ receptor antagonist losartan on cell proliferation activity.After treatment with Ang Ⅱ and piperine, the cell proliferation activity, the cell cycle distribution and the cell migration were detected by MTT, flow cytometry and Transwell assay respectively.ERK1/2 inhibitor PD98059 and losartan were then applied to determine the expression of cyclin D1, MMP-9, p-ERK1/2, ERK1/2, and β-actin proteins by Western blot assay.Results After 24 h culture, Ang Ⅱ treatment promoted the cell proliferative activity in rat ASMCs (P<0.05), and the promotive effect of 10-7 mol/L Ang Ⅱ was the most significant.Additionally, losartan blocked the Ang Ⅱ-induced cell proliferative activity in rat ASMCs (P<0.05).10-7 mol/L Ang Ⅱ treatment resulted in the elevated cell proliferative activity, higher S phase fraction, increased migrated cell number, and enhanced expression of cyclin D1, MMP-9and p-ERK1/2 proteins (P<0.05);these effects were dose-dependently reversed by piperine.Both PD98059 and losartan blocked Ang Ⅱ-induced expression of p-ERK1/2, cyclin D1 and MMP-9 proteins in rat ASMCs.Conclusions Piperine may inhibit Ang Ⅱ-induced cell proliferation and cell migration via ERK1/2 signaling pathway in rat ASMCs.

The clinical and pathological features of 85 cases with positive cerebrospinal fluid cytology by thin-layer centrifugal cytological test / 中华内科杂志

Objective To evaluate cytological test of cerebrospinal fluid in the diagnosis of meningeal dissemination of tumor cells.Methods The clinical and imaging features of 85 cases with tumor cells diagnosed by thin-layer centrifugal cytological test of cerebrospinal fluid were retrospectively reviewed.The characteristics of cellular morphology and immunocytochemical staining were analyzed.Results The main presentations of all the patinets was meningeal irritation and neurological dysfunction.The features of the brain MRI were meningeal thicking and enhancement,intracranial abnormal signals and intracranial space occupying lesion in part of the patients.Atypical cells were found in 84 cases (98.8％) with the first sample test and immunocytochemical staining was conducted in 48 cases to identify the tissue origin.Meningeal carcinomatosis was shown to be the majority with lung cancer as the dominated tissue type and adenocarcinoma as the most common histological type.Others were lymphatic hematopoietic system (13 cases),melanomas (5 cases),primitive neuroectodermal tumor (3 cases) and glioma (1 case).In addition,12 cases were only proved to be cancer by cytological test of cerebrospinal fluid.Conclusion The thin-layer centrifugal cytological test of cerebrospinal fluid has a relatively high accuracy for detecting disseminated tumor cells of meninges and could be of great help to identify the source and type of lesion with immunocytochemical staining.

SGK1-induced imbalance of T lymphocyte subsets in childhood asthma / 国际儿科学杂志

Objective To study the expression of SGK1 in T lymphocytes from pediatric asthma,and the effect of SGK1 on the differentiation of T cells,also to explore the function of SGK1 regulating the differen-tiation of T subset in pediatric asthma. Methods Twenty-eight children with asthma were recruited in Xi′an children′s hospital and divided into moderate group and severe group according to diagnostic guideline of asth-ma. The serum levels of IL-4,IL-13 and IL-17A were analyzed by ELISA. The CD4 +T cells from PBMC and na?ve T cells were selected using magnetic beads. Na?ve T cells were differentiated in vitro under cytokines. SGK1 expression were analyzed with Real-time PCR. The ability of Th2 and Th17 on secreting IL-4 and IL-17A were detected after SGK1 was inhibited by siRNA. In vivo,shRNA-SGK1 Na?ve T cells were transferred into the mice asthma models by intravenous injection. The airway inflammation were observed in shRNA-SGK1 Na?ve T models. Results Compared with healthy children,the serum levels of IL-4、IL-13 and IL-17A increased signifi-cantly in the children with asthma. Importantly,the levels of these three cytokines were much higher with the de-velopment of asthma. SGK1 were up-regulated remarkably in CD4 +T cells from the children with asthma and were positively correlated with IL-13 and IL-17A. Besides,SGK1 expression increased in the differentiated Th2 and Th17 in vitro,but had no change in the differentiated Th1. The levels of IL-4 and IL-17A associated with Th2 and Th17 decreased after SGK1 was inhibited by siRNA. Similarly,In vivo,the serum levels of IL-13 and IL-17A and airway inflammation were reduced in shRNA-SGK1 Na?ve T models. Conclusion The over-expres-sion of SGK1 in pediatric asthma enhances the asthma progress by promoting the differentiation of T subsets.

Expression of autophagy-related proteins in malformations of cortical development / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the expression of autophagy-related proteins (Beclin-1, LC3 and p62) in brain tissue with malformations of cortical development and related molecular pathogenesis.</p><p><b>METHODS</b>The brain tissue of 18 cases with epileptogenic foci resection, including 6 cases of tuberous sclerosis complex (TSC), 6 cases of focal cortical dysplasia type IIb (FCD IIb) and 6 cases of focal cortical dysplasia type I (FCD I), were retrieved. Immunohistochemical study for Beclin-1, LC3 and p62 proteins was performed. The degree of positivity for Beclin-1 and LC3 proteins was compared. Western blot was used to quantitatively analyze the LC3 protein in focal lesion of each disease groups.</p><p><b>RESULTS</b>Immunohistochemical study showed that the three proteins were mainly expressed in the dysmorphic neurons and balloon cells/giant cells of TSC and FCD IIb. The positivity was more intense in the dysmorphic neurons than the other cell types. Immunostaining for Beclin-1 showed granular or diffuse cytoplasmic positivity, in addition to the strong expression in axons. On the other hand, LC3 showed diffuse or perinuclear cytoplasmic expression. The staining for p62 was mainly cytoplasmic or perinuclear and sometimes nuclear. In FCD type I, only individual cells showed positive expression for the three proteins. The number of Beclin-1 and LC3-positive cells was larger in TSC group, followed by FCD IIb group and FCD I group.And there were significant differences between TSC group and FCD I group, as well as FCD IIb group and FCD I group (P<0.05). Quantitative expression of LC3 protein by Western blot showed smaller amount in TSC group, followed by FCD IIb group and FCD I group.</p><p><b>CONCLUSIONS</b>The dysmorphic neurons and balloon cells/giant cells of TSC and FCD IIb show abnormality in autophagy, resulting in intracytoplasmic protein accumulation. There are differences in molecular pathogenesis in these cell types.</p>

Investigation on malnutrition status among the elderly in age care institutions in Shanghai suburb and its influential factors / 中华老年医学杂志

Objective To investigate the malnutrition status among the elderly in age care institutions in Shanghai suburb and analyze its potential influential factors.Methods The MiniNutritional Assessment (MNA) was adopted to evaluate the nutritional status of the 190 elderly people in age care institutions.The dietary supply by the institution canteen and the quantity of residual food left by the malnourished elderly people were weighted.Results In the age care institutions,the malnutrition rate reached 23.7％,47.9％ of the elderly people were at the risk of malnutrition,and only 28.4％ of the elderly people were well nourished.Logistic regression analysis showed that the major influential factors for malnutrition in the elderly people were food intake ability,mobile capability,chewing and swallowing ability,ageing and mental Illness.The malnourished elderly people had the most residual meat and vegetables,and insufficiency of nutrient intake was the main cause for the malnutrition in the elderly people.Conclusions The elderly people in age care institutions in Shanghai suburb have the higher risk of malnutrition.The malnutrition occurs under influence of many factors,of which some are unavoidable,however,some factors like dietary factors can be changed to improve the nutritional status of the elderly people in age care institutions.

Screening of novel miRNAs targeting EZH2 3' untranslated region using lentivirus miRNAs library and their expressions in breast cancer cells and tissues / 南方医科大学学报

<p><b>OBJECTIVE</b>To screen novel miRNAs targeting EZH2 3' untranslated region (UTR) in recombinational MCF-7 breast cancer cells over-expressing EZH2 3' UTR and quantitative analyze the expressions of the screened miRNA in breast cancer cells and tissues.</p><p><b>METHODS</b>A lentiviral library was transfected into the recombinant cell line MCF-7. The cells were screened with cytotoxic agents before extraction of the genome for amplification of the miRNA precursors using PCR. The screened miRNAs were identified with sequence analysis and their expressions were analyzed quantitatively with real-time PCR in breast cancer cells and tissues.</p><p><b>RESULTS</b>Seven miRNAs were screened from the recombinant MCF-7 cells, namely miR-15b, miR-16-2, miR-181b2, miR-217, miR-224, miR-329-1, and miR-487b, all of which failed to be predicted by bioinformatics software. Real-time PCR showed that miR-217, miR-329-1, and miR-487b were over-expressed in MCF-7 cells, and the expression of miR-15b and miR-16-2 was significantly increased in cancer tissues compared with the adjacent tissues (P<0.05).</p><p><b>CONCLUSION</b>Novel targeted miRNAs that can not be predicted by bioinformatics software were successfully screened from MCF-7 breast cancer cells over-expressing EZH2 3' UTR. These miRNAs are expressed differentially between normal breast cells and breast cancer tissues.</p>

Comparison of effects of dexmedetomidine and fentanyl on median effective concentration of propofol required for inducing respiratory depression / 中华麻醉学杂志

Objective To compare the effects of dexmedetomidine and fentanyl on the median effective concentration (EC50) of propofol required for inducing respiratory depression.Methods ASA physical status Ⅰ or Ⅱ patients,aged 30-50 yr,weighing 50-75 kg,with body mass index of 22-28 kg/m2,scheduled for elective gynecological surgery under general anesthesia,were randomly divided into 3 groups:control group (group C),dexmedetomidine group (group D) and fentanyl group (group F).Dexmedetomidine 0.5 μg/kg and fentanyl 1.5 μg/kg in 10ml of normal saline were infused over 10 min in D and F groups,respectively,while the equal volume of normal saline was given in group C.Lidocaine was injected intravenously followed by target-controlled infusion of propofol.In C,D and F groups,the initial target concentration of propofol was 2.5,2.1 and 1.1 μg/ml,respectively,and the ratio between the two successive concentration gradients was 1.1.It was defined as positive when patients developed respiration depression.EC50 and 95％ confidence interval of propofol inducing respiratory depression were calculated.BIS values and OAA/S scores were recorded after admission to operating room (T0),after infusion of dexmedetomidine,fentanyl or normal saline,at 5 and 10 min after the start of propofol target-controlled infusion and after the target effect-site and plasma concentrations were balanced.Results EC50 (95％ confidence interval) of propofol required for inducing respiratory depression was 2.72 (2.55-2.91),2.15 (2.05-2.27)and 1.17(1.08-1.25) μg/ml in C,D and F groups,respectively.Compared with group C,the EC50 was significantly decreased in D and F groups and the BIS values and OAA/S scores were increased in group F (P ＜ 0.05),and no significant changes were found in the BIS values and OAA/S scores in group D (P ＞ 0.05).The EC50 was significantly decreased and the BIS values and OAA/S scores was increased in group F as compared with group D (P ＜ 0.05).When the BIS values reached 65 or OAA/S scores was 3,the effect-site concentration was significantly lower in D and F groups than in group C (P ＜ 0.05).In C,D and F groups,the percentage of patients who developed upper airway obstruction caused by glossocoma was 100％,60％ and 20％,respectively,and who developed decreased respiratory rate or apnea (RR≤ 8 bpm or respiratory arrest time≥ 15 s) was 0,40％ and 80％,respectively.Conclusion Although dexmedetomidine induces no respiratory depression,dexmedetomidine can enhance the potency of propofol required for inducing respiratory depression and the prtency is lower than that of small-dose fentanyl.

Comparison of multi-slice spiral CT versus 3.0 T MR imaging in assessing tumor regression after radiofrequency ablation of hepatocellular carcinomas / 中华放射学杂志

Objective To assess the diagnostic value of MR imaging in follow-up evaluation of patients with hepatocellular carcinomas ( HCC ) treated with radiofrequency ablation ( RFA ) and to compare it with that of multi-slice CT.Methods From December 2009 to September 2011,there were 48 patients (56 HCCs) treated with RFA after transcatheter arterial chemoembolization (TACE). MR imaging and multi-slice CT were performed for follow-up.Two radiologists independently reviewed these images,detection of residual or recurrent tumor were assessed on a five-point scale and compared with Kappa test and with the method of receiver operating characteristic (ROC) curve analysis.Sensitivity,specificity and accuracy were evaluated.Results The observer agreement rate for MR imaging was higher ( 0.925 ) than for multi-slice CT (0.701,P ＜ 0.05).The area under the ROC curve (AUC)of MR imaging( 0.987 and 0.971 by two radiologists respectively) was significantly higher than that of CT( 0.674 and 0.598 by two radiologists respectively),P ＜0.05. The sensitivity, specificity and accuracy of detection rate for MRI [100％(22/22),95.5％ (86/90) and 95.5％ ( 107/112),respectively] were significantly different with that for multi-slice CT [40.9％ ( 9/22 ),57.8％ ( 52/90 ) and 60.7％ ( 68/112 ),respectively]. Conclusion Diagnostic accuracy and detection rate of residual or recurrent tumor were found to be superior with MR imaging than with multi-slice CT.

The displacement of silver clips in breast cavity during kV-plain film guided three-dimensional con-formal external-beam partial breast irradiation assisted by active breathing control / 中华放射肿瘤学杂志

Objective To measure the displacement of the silver clips guided by kV-plain film at state of moderate deep inspiration hold(mDIBH) assisted by active breathing control(ABC) and to explore the margin of clinical target volume(CTV) to planning target volume(PTV) for breast cancer patients treated with three-dimensional conformal external-beam partial breast irradiation (EB-PBI) assisted by ABC. Methods The patients undertook CT simulation assisted by ABC to get the CT images on the respiratory condition of mDIBH. Four selected silver clips in breast cavity were delineated and the cavity based on all of the clips were delineated as gross tumor volume (GTV). Before each irradiation, two orthogonal kV-plain films were taken for the patients in the respiratory condition of mDIBH assisted by ABC device. 2D-2D auto-matie registration was performed based on pixel between the kV-plain films and the digital reconstructed radi-ographs(DRR). Then manual registration was undertook to get the shifts of the four clips separately at LAT, LNG,and VRT directions. Based on the shift data,the margins of CTV to PTV at LAT,LNG and VRT direc-tions were calculated. Results The margins from CTV to PTV were 5.00 mm,7.78 mm and 9.30 mm at LAT,LNG and VRT directions based on the clip at cephal border of the cavity. The corresponding margins were 4.40 mm,6.43 mm and 6.73 mm based on the clip at bottom of the cavity;5.04 mm,8.63 mm and 10.54 mm based on the clip at lateral border of the cavity;5.40 mm,8.59 ram and 10.81 mm based on the clip at pedal border of the cavity. Conclusions The silver clips in breast cavity can be clearly showed on the kV-plain film. The displacement of the clips can be exactly measured by registration of kV-plain film and planning DRR in condition of mDIBH assisted by ABC. The margins from CTV to PTV for EB-PBI can be calculated based on the displacement of the clips.