1.Study on Inhibitive Mechanism of Epithelial-mesenchymal Transition of Trophoblast Cells by Targeted Silencing of Notch-1 Gene
Yang YU ; Leilei WANG ; Tao SHANG
Journal of China Medical University 2015;(3):238-242,258
Objective To discuss the effect and related mechanism on epithelial?mesenchymal transition(EMT)process and invasive ability of the trophoblasts JEG?3 by targeted silencing of Notch?1 gene. Methods Notch?1?siRNA interference fragment was designed and synthesized to sta?bly transfect JEG?3 cells. Changes in the invasive ability of trophoblasts were detected by Transwell chamber invasion assay. The expressions of Notch?1 ,E?cadherin ,vimentin and EMT transcriptive regulators slug ,snail ,twist were analyzed in transfected trophoblasts by Western blot. Results The relative expression of Notch?1 protein in JEG?3 and the invasive ability of JEG?3 were significantly decreased after transfection with Notch?1?siRNA,the expression of the epithelial indictor E?cadherin was significantly increased while the mesenchymal indicator vimentin was de?creased with the reduction of EMT transcriptive regulators slug,snail and twist. Conclusion The invasion of trophoblasts was affected by targeted si?lencing of Notch?1 gene and Notch?1 can reduce the invasion of trophoblasts through influencing EMT process.
2.Carcino-embryonic antigen targeted and drug loaded ultrasound nanoparticle agents inhibit growth of ovarian cancer cells in vitro
Hang ZHOU ; Xiaoling HUANG ; Pan LI ; Tingting SHANG ; Leilei ZHU ; Zhigang WANG
Chinese Journal of Medical Imaging Technology 2017;33(6):816-821
Objective To prepare carcino-embryonic antigen (CEA) targeted and paclitaxel loaded phase-shifting PLGA nanoparticles (Ab-PTX-NPs),and investigate the targeting capability and inhibition to the ovarian cancer cell in vitro.Methods Single-emulsion/solvent evaporation (O/W) and carbodiimide method were used to prepare the Ab-PTX-NPs.The size of nanoparticles was determined by Malvern analyzer.The encapsulation and drug loaded efficiency of paclitaxel were detected by high performance liquid chromatography.And the drug release characteristics was measured by dialysis method in constant temperature shaker.The targeting ability of Ab-PTX-NPs to the ovarian cancer SKOV3 cell was evaluated by the laser scanning confocal microscope and flow cytometry.And the inhibition ability of Ab-NPs was investigated by the CCK-8 assays.Results The size of Ab-PTX-NPs was (397.70±99.95)nm.The encapsulation efficiency and drug loading capacity of PTX were (67.26±4.15) % and (6.31±0.39) %,respectively.The conjugating rate of Anti-CEA antibody was (92.74 ± 5.75) %.The targeting study in vitro showed that such a number of contrast agents landed around the SKOV3 cells in targeting group,and the mean fluorescence intensity of ovarian cells in targeting group was significantly higher than other groups (P<0.05).After 24 h,the viability rate of ovarian cells in targeting group was lower than the non-target group (P<0.05),only higher than that of the pure PTX group (P<0.05).But there was no significant difference between the targeting group and the pure PTX group (P>0.05) at 48 h.Conclusion The CEA targeted and paclitaxel loaded phase-shifting PLGA nanoparticles are successfully prepared.It can enhance ultrasound imaging well after activated by LIFU.With high drug-loading efficiency and fast drug release velocity,the Ab-PTX-NPs appeares great targeted ability.
3.Prediction of the onset time of acute stroke by deep learning based on DWI and FLAIR
Liang JIANG ; Leilei ZHOU ; Zhongping AI ; Yuchen CHEN ; Song'an SHANG ; Siyu WANG ; Huiyou CHEN ; Mengye SHI ; Wen GENG ; Xindao YIN
Chinese Journal of Radiology 2021;55(8):811-816
Objective:To evaluate the effect of deep learning based on DWI and fluid attenuated inversion recovery (FLAIR) to construct a prediction model of the onset time in acute stroke.Methods:A total of 324 cases of acute stroke with clear onset time, from January 2017 to May 2020 in Nanjing First Hospital, were retrospectively enrolled and analyzed. The patients were divided into a training set of 226 patients and a test set of 98 patients according to the complete randomization method using a 7∶3 ratio, and the patients were divided into ≤ 4.5 h and >4.5 h according to symptom onset time in each group. The acute infarction areas on DWI and the corresponding high signal area on FLAIR were manually outlined by physician. Using the InceptionV3 model as the basic model for image features extraction, the deep learning prediction model based on single sequence (DWI, FLAIR) and multi sequences (DWI+FLAIR) were established and verified. Then the area under curve (AUC), accuracy of human readings, single sequence model and multi sequence model in predicting the acute stroke onset time from imaging were compared.Results:DWI-FLAIR mismatch was found in 94 cases (94/207) of patients with symptom onset time from imaging ≤ 4.5 h, while in 28 cases (28/117) of patients with symptom onset time from imaging >4.5 h. ROC analysis showed that the AUC of DWI-FLAIR mismatch in predicting acute stroke onset time from imaging was 0.607, and the accuracy was 60.2%. The prediction model of deep learning based on single sequence showed that the AUC of FLAIR was 0.761 and the accuracy was 71.4%; the AUC of DWI was 0.836 and the accuracy was 81.6%. The AUC of predicting stroke onset time based on the multi-sequence (DWI+FLAIR) deep learning model was 0.852, which was significantly better than that of manual identification ( Z = 0.617, P = 0.002), FLAIR sequence deep learning model ( Z = 2.133, P = 0.006) and DWI sequence deep learning model ( Z = 1.846, P = 0.012). Conclusion:The deep learning model based on DWI and FLAIR is superior to human readings in predicting acute stroke onset time from imaging, which could provide guidance for intravenous thrombolytic therapy for acute stroke patients with unknown onset time.
4. Role of PI3K/Akt signaling pathway in ischemic rats underwent cardiac shock waves therapy
Leilei SHANG ; Zhen SU ; Xuejuan MA ; Yanqiong WANG ; Yu WANG ; Qixian WANG ; Ping YANG
Chinese Journal of Cardiology 2019;47(6):457-464
Objective:
To investigate the role of PI3K/Akt signaling pathway in ischemic rats underwent cardiac shock therapy.
Methods:
Adult male Sprague Dawley (SD) rats weighing 220-250 g were used to establish a heart failure model by ligation of the left anterior descending coronary artery. Rat models were defined by echocardiographic assessment at 4 weeks post operation and heart failure rats were randomly divided into 4 groups,namely heart failure group (HF group, 9 cases),heart failure+cardiac shock waves therapy group (HF+CSWT group, 9 cases),heart failure+inhibitor(HF+LY294002 group, 9 cases),heart failure+cardiac shock waves therapy group+inhibitor (HF+CSWT+LY294002 group, 9 cases),and another 9 sham-operated SD rats served as control group (sham group, 9 cases). At 8 weeks postoperation, echocardiography was used to evaluate cardiac function in each group,myocardial infarct size was measured by TTC staining,the apoptotic index of rats cardiomyocytes were detected by TUNEL method,the myocardial mRNA expression of apoptosis-related factor was detected by real-time quantitative PCR, the protein expression levels of PI3K/Akt signaling pathway and apoptosis-related pathways were detected by Western blot.
Results:
(1) Eight weeks after operation, left ventricular end diastolic diameter (LVEDD) and left ventricular end systolic diameter (LVESD) were significantly lower in HF+CSWT group than in HF group (all