Objective To explore the role of the PI3K/Akt signaling pathway in the calvarial osteolysis induced by TCP wear particles in mice model.Methods Thirty-six male ICR mice were randomly divided into a sham group (n=12),TCP group (n=12)and a LY294002-treated group (n=12).A murine calvarial model of osteolysis was established through implanting 30 mg of TCP particles onto the surface of bilateral parietal bones following the removal of the periosteum.On the second postoperative day,LY294002 (5 mg·kg-1)was locally injected to the calvarium under the periosteum three times a week;mice in the sham group received local injection of normal saline (N.S.)in the calvarium,and the injection time was consistent with that of LY294002.Two weeks later,the calvaria and periostea were obtained after the mice were executed.The calvarial osteolysis,bone mineral density (BMD)and bone mineral content(BMC)were analyzed using Micro-CT,Hematoxylin-Eosin (HE)staining was conducted to observe the inflammatrory response and formation of osteoclasts.Real-time PCR was applied to detect the mRNA level of tartrate-resistant acid phosphatase (TRAP),the marker of osteoclasts formation,cathepsin K (CstK),receptor activator for nuclear factor-κB kigand (RANKL)and c-Fos.The release of tumor necrosis factor-α (TNF-α),interleukin-6 (IL-6)and IL-1β were measured using enzyme-linked immumsorbent assay (ELISA).Results Micro-CT and histological analysis indicated that LY294002,the specific inhibitor of PI3K,significantly prevented TCP wear particles-induced osteolysis and osteoclastogenesis,and increased BMD and BMC in the calvaria of mice.Real-time PCR data revealed LY294002 significantly suppressed the increase in mRNA level of osteoclastogenic genes such as TRAP,CstK,RANKL and c-Fos in the calvaria of TCP wear particles-implanted group.ELISA assay showed that TCP wear particles-induced release of TNF-α,IL-1β and IL-6 was significantly inhibited by LY294002 treatment.Furthermore,LY294002 significantly attenuated TCP wear particles-triggered activation of Akt,and down-regulated the level of p-AktSer473 and p-AktThr308.Conclusion PI3K/Akt signaling pathway contributes to TCP wear particle-induced osteolysis,and can be developed as a new therapeutic target for the prevention and treatment of bone destruction diseases caused by wear debris.

OBJECTIVETo investigate the effects of Annexin A2 (ANXA2) deficiency on the malignant biological behaviour of hepatoma cells.

METHODSThe human hepatocellular carcinoma (HCC) cells lines MHCC97-H, HepG2, SMMC-7721, SMMC-7402 and L02 were evaluated. The expression and distribution of ANXA2 were analysed by western blotting, real-time PCR, immunofluorescence and immunohistochemistry.Cell cycle was assessed by flow cytometry and propidium iodide staining. Effects of ANXA2 silencing on invasion and migration potential were assessed by transwell assay and wound healing assay, respectively. Proliferative potential was assessed by CCK-8 kit in vitro and xenograft tumour-growth assay in vivo. The t-test, chi square test, rank sum test, q-test and F-test were used for statistical analyses.

RESULTSThe expression level of ANXA2 was markedly higher in the MHCC97-H cells with high metastasis potential than in the HepG2, SMMC-7721, SMMC-7402 and L02 cells. The efficiency of shRNA-mediated ANXA2 deficiency was more than 80%. Immunofluorescence analysis of the MHCC97-H cells indicated that ANXA2 expression was mainly localized to the cellular membrane and cytoplasm, with some nuclear localization. Down-regulation of ANXA2 led to S-phase arrest of HCC cells (q =8.001, P =0.002) and an inhibition of proliferation (q =17.140, P less than 0.01), migration (q =12.808, P less than 0.01) and invasion potential (q =9.069, P =0.002). Xenograft tumour-growth assay indicated that shRNA targeting of ANXA2 led to lower tumour weight (q =11.968, P < 0.001) and down-regulated ANXA2 expression (Z =2.530, P =0.011).

CONCLUSIONDown-regulation of Annexin A2 gene transcription effectively changes the biological behaviours of hepatoma cells, and may represent a potential target of HCC molecular therapies.

Animals ; Annexin A2 ; genetics ; Carcinoma, Hepatocellular ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Humans ; Liver Neoplasms ; pathology ; Neoplasm Transplantation ; RNA, Small Interfering ; genetics ; Signal Transduction ; Transcription, Genetic

Objective: To analyze the failure patterns of locoregional recurrence (LRR) and investigate the range of radiotherapy in T_1-2N₁ breast cancer patients undergoing modified radical mastectomy. Methods: From September 1997 to April 2015, 2472 women with T_1-2N₁ breast cancer after modified radical mastectomy without neoadjuvant systemic therapy were treated in our hospital. 1898 patients who did not undergo adjuvant radiotherapy were included in this study. The distribution of accumulated LRR was analyzed. The LR and RR rates were estimated by the Kaplan-Meier method, and the prognostic factors were identified in univariate analyses with Log-rank test. Multivariate analysis was performed using Cox logistic regression analysis. Results: With a median follow-up of 71.3 months (range 1.1-194.6), 164 patients had LRR, including supraclavicular/infraclavicular lymph nodes in 106(65%), chest wall in 69(42%), axilla in 39(24%) and internal mammary lymph nodes (IMNs) in 19 patients (12%). In multivariate analysis, age (>45 years vs.≤45 years), tumor location (other quadrants vs. inner quadrant), T stage (T₁ vs. T₂), the number of positive axillary lymph nodes (1 vs. 2-3), hormone receptor status (positive vs. negative) were significant prognostic factors for both LR and RR. Conclusions In patients with T_1-2N₁ breast cancer after modified radical mastectomy, the most common LRR site is supraclavicular/infraclavicular nodal region, followed by chest wall. The axillary or IMN recurrence is rare. The prognostic factors for LR and RR are similar, which indicates that supraclavicular/infraclavicular and chest wall irradiation should be considered for postmastectomy radiotherapy.

Objective To study the effect of deep inspiration breath-hold(DIBH)technique on the heart dose in whole breast irradiation(WBI)for left breast cancer after breast-conserving surgery, and to investigate the anatomical factors for heart dose. Methods Fifteen patients with left breast cancer who received WBI after breast-conserving surgery and met breathing control requirements were prospectively enrolled as subjects. Simulated CT scans were performed during free breathing(FB)and DIBH. The WBI plans were optimized based on DIBH images.The position,volume,and radiation doses to the heart and lung were compared between the status of FB and DIBH. Correlation of heart dose with various anatomical factors was analyzed in FB status. Between-group comparison of categorical data was made by nonparametric Wilcoxon rank test.A two-variable correlation analysis was made by the Pearson method.Results There was no significant difference in heart volume between the status of FB and DIBH(P=0.773).The volume of both lungs was significantly larger in DIBH status than in FB status(P=0.001). The mean and maximum doses and V5-V40for the heart,left anterior descending coronary artery,left ventricle,right ventricle,and left lung were significantly lower in DIBH status than in FB status(all P<0.05). The greater DIBH increased the lung volume,the greater the mean heart dose decreased. In FB status,the left breast volume,heart-to-lung volume ratio,distance between the inferior margins of breast and heart,and maximum heart margin distance showed a linear correlation with heart dose. Particularly, the heart-to-lung volume ratio and maximum heart margin distance were independently correlated with heart dose. Conclusions DIBH technique in WBI for left breast cancer after breast-conserving surgery significantly reduces heart and lung doses compared with FB. Changes in lung volume are the basis for improving the relative anatomical location of the heart. The heart-to-lung volume ratio and maximum heart margin distance may provide a reference for DIBH technique.

Objective To analyze the correlation between treatment time and radiotherapy plan of deep inspiration breath-hold (DIBH) technique for the whole breast irradiation (WBI) in the left breast cancer after breast-conserving surgery,verify the inter-fractional reproducibility of radiotherapy,observe the heart location and dosimetric changes and calculate the effect of DIBH upon the WBI setup error after the surgery.Methods We prospectively enrolled 15 patients with left breast cancer undergoing WBI after breast-conserving surgery,who met the requirement of D1BH.Treatment time was recorded,its correlation with the number of field and monitor unit was analyzed.Inter-fractional setup errors and PTV delineation were calculated using cone beam CT (CBCT).The accuracy of the position and dose of the heart during radiotherapy was verified by the imaging fusion of CBCT and CT images.The variables among groups were analyzed by non-parametric Firedman test.Results The average treatment time of DIBH radiotherapy was 4.6 minutes.The treatment time was correlated with the maximal and total number of sub-fields and total monitor units.During DIBH treatment,the mean cardiac displacement volume was 19.1 cm3(3.8％).The mean cardiac dose difference between CBCT and planning CT was 5.1 cGy,and there was no significant difference in the heart V5-V30.The mean inter-fractional system setup error (∑) and random setup error (σ) in the left-right (x),superior-inferior (y) and anterior-posterior (z) direction were ∑x 1.9 mm,∑y 2.1 mm,∑z 2.0 mm,σx 1.3 mm,σy 1.3 mm,σz 1.4 mm,respectively.The corresponding minimal margins for setup error were 5.7 mm,6.2 mm and 6.0 mm,respectively.Conclusion DIBH for WBI after breast-conserving surgery does not significantly prolong the treatment time.Treatment time is related to treatment plan.DIBH yields high inter-fractional reproducibility and protects the heart.

Objective To quantify the setup errors measured with kV cone-beam CT (CBCT) using breast board with or without a thermoplastic head mask in breast cancer patients who received whole breast and supraclavicular nodal region irradiation.Clinical target volume (CTV) to planning target volume (PTV) in 3 directions were also calculated.Methods The study included thirty patient receiving both whole breast and supraclavicular nodal region irradiation on Elekta Versa HD linear accelerators (Elekta Oncology Systems,Crawley,UK) between June 2016 and January 2018.The setup error data were retrospectively analyzed.All the patients were immobilized in the supine position on a breast board system (Carbon fibre breast board,Civco,Iowa,USA) with both arms raised.Twenty of the patients added an extra thermoplastic head mask to immobilize the neck.A CBCT scan was taken before treatment at the first 3 to 5 fractions and then once every week.Registration with the planning CT was performed and adjusted to match the target volume of the supraclavicular nodal region and the cervical vertebra by experienced medical staff,the position of the arms and the breast were also considered.For all patients the couch shifts in left-right (x),superior-inferior (y),anterior-posterior (z) were recorded.Statistical analysis included two-tails significance tests (t-Student and Manne-Whitney test for means,x2 test for variances).Population of the two groups was compared in terms of distribution of the mean shift (systematic error) and their standard deviations (random error).Van Herk's setup margin defined as MPTV =2.5Σ + 0.7δ was calculated in patients with or without mask immobilization.Results Altogether,56 images and 109 images were acquired in breast board only group and head mask group,respectively.Shifts of the breast board only group and the head mask group in x,y,z were 0.212±0.174 cm vs.0.272±0.242 cm (P=0.070);0.364±0.246 cm vs.0.242±0.171 cm (P=0.001);0.423±0.302 cm vs.0.364±0.269 cm (P=0.204).Proportion of the shift less than 5 mm in the breast board only group and the head mask group were 91.07％ vs.85.32％ (P=0.294);67.86％ vs.89.91％ (P=0.001);67.86％ vs.74.31％ (P=0.381).The CTV to PTV margin in x,y,z were 0.645 cm,0.981 cm,1.317 cm in breast board only group and 0.873 cm,0.709 cm,0.961 cm in head mask group,respectively.Setup error in the x direction was significantly correlated with BMI (P=0.001).Conclusions For the alignment and immobilization of patients who received whole breast and supraclavicular nodal region irradiation,using a breast board in combination of a thermoplastic head mask may significantly help to reduce the shift variance in superior-inferior direction compared to using breast board only.The anterior-posterior error is relatively large,other immobilization device or patient alignment methods are needed to be further explored to improve the accuracy.

Objective: To validate whether the prognostic stage groups by the 8th edition of the American Joint Committee on Cancer (AJCC) staging system provides improved prognostic accuracy in T1-2N1M0 postmastectomy breast cancer patients compared to 7th edition. Methods: a total of 1 823 female patients with T1-2N1M0 breast cancer who underwent mastectomy and axillary lymph node dissection without neoadjuvant chemotherapy were analyzed and restaged according to 8th edition. Univariate analysis of prognostic factors was evaluated by using log-rank test. Multivariate analysis was estimated by using the Cox proportional hazards model. The prognostic accuracy of the two staging systems was compared using receiver operating characteristic (ROC) analyses and the concordance index (C-index). Results: 5-year locoregional recurrence rate (LRR) for the whole group was 6.0%, 5-year distant metastasis (DM) rate was 11.5%, 5-year disease-free survival (DFS) was 85.0%, and 5-year overall survival (OS) was 93.1%. Cox analysis showed that 7th edition of the AJCC staging system and progesterone receptor status were independent risk factors for LRR, DM, DFS and OS (P<0.05). Compared with stage by 7th edition, 1 278(70.1%) were assigned to a different prognostic stage group: 1 088 (85.1%) to a lower stage and 190 (14.9%) to a higher stage. LRR, DM, DFS and OS were significantly different between prognostic stage ⅠA, ⅠB, ⅡA, ⅡB and ⅢA according to 8th edition of the AJCC staging system(P<0.001). Prognostic stage had significantly higher C-indexes and provided better estimation of prognosis compared to stage by 7th edition of the AJCC staging system (P<0.001). Conclusion The prognostic stage groups of 8th edition AJCC staging system has superior prognostic accuracy compared to 7th edition in T1-2N1M0 breast cancer, and has better clinical therapeutic guidance value.

Objective:To analyze the efficacy of chest wall boost radiotherapy in stage T 4 breast cancer patients after modified radical mastectomy. Methods:A retrospective analysis was performed on the data of 148 stage T 4 breast cancer patients who were admitted from 2000 to 2016 and received radiotherapy after modified radical mastectomy. There were 57 cases in the chest wall boost radiotherapy group and 91 cases in the conventional dose group. Radiotherapy was performed by conventional+ chest wall electron beam, three-dimensional conformal+ chest wall electron beam, intensity modulated radiotherapy+ chest wall electron beam irradiation. EQD 2 at the boost group was >50Gy. All patients received neoadjuvant chemotherapy. Kaplan-Meier method was used to analyze survival; Logrank was used to test differences; and Cox model was used to do multivariate prognostic analysis. Results:The median follow-up time was 67.2 months. The 5-year rates of chest wall recurrence (CWR), locoregional recurrence (LRR), disease-free survival (DFS), and overall survival (OS) were 9.9%, 16.2%, 58.0%, and 71.4%, respectively. The 5-year rates of CWR, LRR, DFS, and OS with and without chest wall boost radiotherapy were 14% vs. 7%, 18% vs. 15%, 57% vs. 58%, 82% vs. 65%( P>0.05), respectively. Multivariate analysis showed that chest wall boost radiotherapy had no significant effect on prognosis ( P>0.05). Among 45 patients in the recurrent high-risk group, boost radiotherapy seemed to have higher OS rate ( P=0.058), DFS rate ( P=0.084), and lower LRR rate ( P=0.059). Conclusions:Stage T 4 breast cancer patients had strong heterogeneity. Chest wall boost radiotherapy did not apparently benefit all patients. For patients with 2-3 high risk factors including positive vascular tumor embolus, pN 2-N 3, and hormone receptor negative, chest wall boost radiotherapy showed a trend of improving efficacy.

ObjectiveTo explore the effect of external diaphragm pacing therapy combined with abdominal functional electrical stimulation on respiratory function for stroke patients. MethodsFrom October, 2020 to September, 2022, 54 stroke patients were randomly divided into control group (n = 18), external diaphragm pacing group (n = 18) and combined treatment group (n = 18). All the groups received breathing training, while the external diaphragm pacing group received external diaphragm pacing therapy, and the combined treatment group received external diaphragm pacing and abdominal functional electrical stimulation therapy, for two weeks. They were measured forced vital capacity (FVC), forced expiratory volume in first second (FEV₁), ratio of forced expiratory volume in first second in forced vital capacity (FEV₁/FVC), peak expiratory flow (PEF), maximal inspiratory pressure (MIP) and maximal expiratory pressure (MEP) with pulmonary function instrument; measured diaphragmatic excursion (DE) and diaphragmatic thickness (DT) with ultrasound, before and after treatment. ResultsThree cases in the control group, two cases in the external diaphragm pacing group and one case in the combined treatment group dropped off. The FVC, FEV₁, PEF, MIP, MEP and DE improved in all the groups (|t| > 3.366, P < 0.01) after treatment; and the FVC, FEV₁, MIP and DE increased more in the combined treatment group and the external diaphragm pacing group than in the control group (P < 0.05); the FVC and FEV₁ increased more in the combined treatment group than in the external diaphragm pacing group (P < 0.05). ConclusionExternal diaphragm pacing therapy may improve ventilation and inspiratory muscle strength, and increase diaphragm movement for stroke patients; while the ventilation improved more after combining with abdominal functional electrical stimulation.

OBJECTIVES: To investigate the clinical characteristics and risk factors for severe events of coronavirus disease 2019 (COVID-19) in elderly patients. METHODS: Retrospective analysis was performed on the clinical data of all elderly COVID- 19 patients treated in Changsha Public Health Treatment Center from January 17, 2020 to March 15, 2020, which included basic diseases, symptoms, test results, and other clinical characteristics, and prognostic indicators such as severity of illness, length of hospital stay, virus shedding time and mortality rate. The differences in clinical characteristics and prognostic indicators between elderly, middle-aged, and young COVID-19 patients were also analyzed. Logistic regression model was used to conduct univariate and multivariate analysis of risk factors for developing severe events in elderly COVID-19 patients; receiver operating characteristic (ROC) curve analysis was used to evaluate the prediction efficacy. RESULTS: Of the 230 COVID-19 adult patients, 34 were young patients (14.8%), 136 were middle-aged patients (59.1%), and 60 were elderly (26.1%). Among the 60 elderly patients, 23 were male (38.3%) and 37 were female (61.7%), with a medium age of 66 years old. Common symptoms were fever (66.7%), cough (50.0%), and fatigue (41.7%). C reactive protein (CRP) was increased significantly. The proportion of severe cases was 31.7%, and mortality was 1.7%. The median length of hospitalization and median virus shedding time were 18.5 days and 21 days, respectively. Compared with the young and the middle-aged patients, the elderly had a higher proportion of hypertension, diabetes, and cardiovascular diseases, more common shortness of breath, higher proportions of pneumonia and severe cases (all <0.05), and the decreased lymphocyte count and lymphocyte percentage (both <0.05), as well as higher CRP and erythrocyte sedimentation rate (ESR) levels (both <0.05). Compared with non-severe cases, severe elderly patients demonstrated higher CRP and aspartate aminotransferase (AST) levels (all <0.05), the reduced lymphocyte count (<0.05), and the prolonged length of hospitalization and virus shedding duration (both <0.05). Univariate logistic regression analysis indicated that the lymphocytes proportion, CRP and AST levels were significantly correlated with the risk for developing severe events in elderly COVID-19 patients (all <0.05). Multivariate logistic regression found that severe events in elderly patients with COVID-19 were significantly correlated with CRP level (OR=1.041, =0.013). ROC curve analysis revealed that the area under the curve (AUC) for CRP to diagnose severe events in elderly COVID 19 patients was 0.851. CONCLUSIONS The proportion of severe cases in elderly COVID-19 patients is higher than that in young and middle-aged patients. CRP level has a good predictive value for the possibility of severe events in elderly COVID-19 patients.
Adult ; Aged ; Betacoronavirus ; C-Reactive Protein ; analysis ; China ; Comorbidity ; Coronavirus Infections ; diagnosis ; physiopathology ; Female ; Humans ; Male ; Middle Aged ; Pandemics ; Pneumonia, Viral ; diagnosis ; physiopathology ; Retrospective Studies ; Risk Factors