2.miR-21 targets Fas ligand-mediated apoptosis in breast cancer cell line MCF-7.
Ming-fu, WU ; Jie, YANG ; Tao, XIANG ; Yan-yan, SHI ; Li-jiang, LIU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2014;34(2):190-4
Over-expression of Fas ligand (FasL) on tumor cell surface can induce the apoptosis of specific activated tumor infiltrating lymphocytes (TILs) via the Fas/FasL pathway, leading to the formation of a site of immune privilege surrounding the tumor mass for escaping immune surveillance and promoting tumor proliferation, invasion and metastasis. The blocking effect of miR-21 on FasL-mediated apoptosis in breast cancers was investigated in this study. The expression levels of miR-21 and FasL in human breast carcinoma cell lines were detected by using RT-PCR and Western blotting. FasL as a target gene of miR-21 was identified by Luciferase assay. The apoptosis of Jurkat T lymphocytes induced by MCF-7 cells was determined by flow cytometry. It was found that in four human breast cancer cell lines, FasL expression level in MCF-7 cells was the highest, while miR-21 was down-regulated the most notably. After miR-21 expression in MCF-7 cells was up-regulated, FasL was identified as a target gene of miR-21. When the effector/target (E/T) ratio of MCF-7 cells and Jurkat cells was 10:1, 5:1 and 1:1, the inhibitory rate of apoptosis of Jurkat T lymphocytes induced by MCF-7 cells was 95.81%, 93.16% and 91.94%, respectively. It is suggested that in breast cancers miR-21 expression is negatively associated with FasL expression, and FasL is a target gene of miR-21. miR-21 targeting and regulating FasL-mediated apoptosis will bring us the possibility of a new tumor immunotherapy via breaking tumor immune privilege.
3.Emodin stimulates glucose uptake by HepG2 hepatocyte through activation of PPAR?
Li-Juan YANG ; Hai-Yan YU ; Yi-Ming MU ; Bao-An WANG ; Jing-Tao DOU ; Ju-Ming LU ; Chang-Yu PAN
Chinese Journal of Endocrinology and Metabolism 1986;0(03):-
Objective To construct PPAR?and PPAR?response element (PPRE)-controlled luciferase expression vectors,and to determine whether the traditional Chinese medicine emodin activates PPAR?and improves the glucose uptake by HepG2 hepatocytes.Methods (1) PPAR?and PPRE luciferase expression vectors were constructed and were applied to screen more than 20 ingredients of the traditional Chinese medicine. (2) HepG2 cells were incubated with emodin which can activate PPAR?and PPRE luciferase activity,and the PPAR?mRNA expression level was evaluated by RT-PCR/Southern blot.(3) PPAR?and glucose transporter 2 (Glut2) proteins were determined by Western blot analysis in HepG2 cells treated with emodin.(4) The glucose uptake rate was measured using 2-deoxy-[~3H]-D-glucose in HepG2 cells after treatment with emodin.Results (1) Emodin stimulated luciferase activity controlled by PPRE in dose-dependent manner at concentrations of 0.04 to 180?mol/L in COS-7 cells.The highest value was about 4 folds of control in the cells treated with 90?mol/L emodin (P
4.The experimental study on infant rabbit lung injury induced by ischemia-reperfusion
Wanshan QIU ; Bing JIA ; Ming YE ; Xiangang YAN ; Gang CHEN ; Qilin TAO ; Sheng SHEN ; Zhanggen CHEN
Chinese Journal of Thoracic and Cardiovascular Surgery 2012;(12):729-731
Objective To explore the characteristics of ischemia-reperfusion induced infant lung damage and the potential mechanisms of the injuried.Methods Both infant (15-21 days old) and adult (5-6 months old) rabbits were subjected to either ischemia-reperfusion or sham operation.Ischemia-reperfusion was induced by clamping the right pulmonary hilum for 1 hour and then removal of the clamp for 4 hours under anesthesia.The lung tissue were sampled for histological examination by light and electron microcopies and for biological evaluation of mitochondrial alterations.Production and expression of free radical species-hydroxyl radical (ROS-HR),malondialdehyde (MDA),superoxide dismutase (SOD),glutathione peroxidase (GSH-PX),myeloid differentiation factor-88 (MyD-88),and nuclear factor-κB (NF-κB) in the lung tissue were also examined.In addition,circulating levels of interleukin-β and tumor necrosis factor-α were measured during the ischemia-reperfusion process.Results In comparison to adult lungs,the infant lungs had more increased neutrophil infiltration,edema,swelled alveolar epithelial and endothelial cells,and severer mitochondrial impairment reflected by damage of the inner membrane as well as decrease in the membrane potential after ischemia-reperfusion.The lungs in infant animals subjected to sham operation displayed higher levels of ROS-HR and MDA and lower levels of SOD and GSH-PX than those in adult controls.The lungs in infants with ischemia-reperfusion were found to further produce more ROS-HR,and MDA,and less SOD and GSH-PX than the ischemia-reperfused adult lungs.Moreover,the circulating levels of interleukin-1β and tumor necrosis factor-α were elevated during the period of ischemia-reperfusion,particularly in the infant animals,which appeared to be associated with the expression of MyD-88 and NF-κB in the lungs.Conclusion Lung ischemia-reperfusion causes more severe lung damage in infants than in adults,probably due to combination of low antioxidant capacity and overproduction of ROS in infants.
5.Relationship between the prevalence of negative life events and depressive and anxiety symptoms and dissatisfaction of school life among middle school students in Shaoxing city, Zhejiang province
Chao XING ; Fangbiao TAO ; Chunyu TU ; Yirong FANG ; Lijun FU ; Yan MA ; Ming LI
Chinese Journal of Health Management 2015;9(6):450-455
Objective To understand the prevalence of negative life events and its association with depressive, anxiety symptoms and dissatisfaction of school life among middle school students in Shaoxing, and to provide scientific evidence for further interventions. Methods A total number of 3 197 students (including 1 134 urban and 2 063 town adolescents) were recruited from 2 middle schools in Shaoxing, their average age was (13.73 ± 1.03) years. The Multidimensional Life Events Rating Questionnaire for Middle School Students (MLERQ), Depression Self-rating Scale for Children (DSRSC) and Screening for Child Anxiety Related Emotional Disorders (SCARED) were used to assess the mental health status, while the school life satisfaction was evaluated by the School Life Satisfaction Rating Questionnaire for Adolescents (SLSRQA), and the data were analyzed by descriptive statistics and multivariate logistic regression models. Results The prevalence of the negative life events was 84.9%, unsatisfactory examination performance (48.6%), unreached the teacher's expectation (46.2%), parental chatter (41.4%), unbalanced learning (37.9%), and getting parents scold (27.6%) constituted the main negative life events. The prevalence of depressive symptom, anxiety symptom, and dissatisfaction of school life were significantly higher among middle school students living with negative life events (41.1%, 22.6%, 26.0%) than those without negative life events (20.9%, 3.5%, 16.5%), (?2=71.33, 94.78, 19.83, P<0.001). The prevalence of the psychosomatic health involvement significantly increased with the increase of the number of the dimensions and events (P<0.001). The multivariate logistic regression showed that negative life events were the risk factors of depression, anxiety and dissatisfaction of school life. Their OR values were 2.483 (1.951-3.160), 7.245 (4.411-11.899) and 1.733 (1.325-2.267), respectively. The risk of occurring mental symptoms among children with the number of dimensions≥4 and/or the number of events≥12 was two times higher than those of children without such increase in numbers of dimensions and events. Conclusion The status of negative life events is serious among middle school students in Shaoxing. There are statistical associations between negative life events and mental symptoms.
6.Endemic fluorosis in Jilin province: analysis of surveillance data for 2006 - 2010
Hai-tao, ZHANG ; Zhen-ming, LU ; Hong-yan, TANG ; Xiu-li, ZHANG ; Lian-ying, FANG
Chinese Journal of Endemiology 2011;30(3):298-302
Objective To identify changes in the occurrence of endemic fluorosis in order to provide scientific basis for making countermeasures. Methods Five villages from 14 counties of mild, moderate and severe fluorosis affected areas were selected by stratified cluster sampling every year in the whole province during 2006 - 2010. Water and urinary fluorine were determined by ion selective electrode method(GB/T 8538-1995); dental fluorosis of children 8-12 years old was diagnosed with Dean method; skeletal fluorosis was diagnosed according to "clinical indexing standards of endemic skeletal fluorosis "(GB 16396-1996), between 2006 and 2008, and "clinical diagnosis standard of endemic skeletal fluorosis"(WS 192-2008) between 2009 and 2010. Results A total of 25 diseased villages were surveyed, 14 with water sources changed, covered a resident population of 8005 people, beneficiary population 7154, and accounting for 89.37% of the resident population; not changed villages 11. In accordance with the "State drinking water health standards", in the 14 changed villages the fluoride in drinking water was qualified (≤ 1.20 mg/L), there were 3 schools whose water fluorine content exceeded the standard; among the 11 villages that did not change water sources 7 drinking water samples fluorine content exceeded the standard. Of the 8 to 12 years old children in villages with changed water sources, 363 of them were checked and 142 dental fluorosis were found, the detection rate of dental fluorosis was 39.12% (142/363); in villages with water sources not changed, 303 children were checked, the detection rate of dental fluorosis was 43.89%(133/303). Of sixteen and elder adults in water source changed villages, 6424 people were checked and 403 skeletal fluorosis were found, skeletal fluorosis detection rate was 6.27% (403/6424); 3572 people were checked in not changed villages, the detection rate of skeletal fluorosis was 13.89%(496/3572). In water sources changed areas, geometric mean of urinary fluoride was in the normal reference value(WS/T 256-2005, 1.40 mg/L)or less. Conclusions Endemic fluorosis is decreased in water improved areas, but in unimproved areas the disease is still severe, and control of endemic fluorosis is still an arduous task.
7.Targeting therapy of choroidal neovascularization by use of polypeptide- and PEDF-loaded immunoliposomes under ultrasound exposure.
Tao, LI ; Ming, ZHANG ; Yong, HAN ; Hong, ZHANG ; Lingjuan, XU ; Yan, XIANG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2010;30(6):798-803
Pigment epithelium derived factor (PEDF) has been proven to be an effective drug for the treatment of choroidal neovascularization (CNV). However, the lack of ideal administration route is the biggest bottleneck preventing PEDF from wider clinical use. In this study, we developed a novel PEDF-carrying system which employed immuno-nano-liposomes (INLs) under ultrasound exposure. PEDF-loaded INLs were prepared by conjugating nanoliposomes to the peptide ATWLPPR specifically targeting the receptor-2 for vascular endothelial growth factor (VEGFR-2) and reversely encapsuling PEDF. RF/6A cells were incubated with PEDF-loaded INLs. CNV models of BN rats were injected with PEDF-loaded INLs. MTT assay was used to evaluate the cytotoxicity of the INLs on RF/6A cells. Flow cytometry was conducted to detect the apoptotic rate of cells. Laser scanning confocal microscopy was employed to observe the binding and transmitting process of PEDF-loaded INLs and to calculate the area of CNV in the rat model. The results showed that the PEDF-loaded INLs could exclusively bind to CNV but not to the normal choroidal vessels. The CNV area was significantly decreased in PEDF treatment groups in comparison with control group (P<0.05). Moreover, PEDF-loaded INLs exposed under ultrasound were more efficient in reducing the CNV area (P<0.05). It was concluded that INLs in combination with ultrasonic exposure can transmit PEDF into cytoplasma with high specificity and efficiency, which strengthens the inhibitory effects of PEDF on CNV and reduces its side effects. PEDF-loaded INLs possibly represent a new treatment paradigm for patients with ocular neovascularization.
8.Contrast-enhanced ultrasonography in evaluation of splenic trauma and injury grading and its clinical apllication
Qiao-rong, LIANG ; Chun-yan, HUANG ; Tong, LIANG ; Shi-ming, TAO ; Zhi-qiang, ZENG
Chinese Journal of Medical Ultrasound (Electronic Edition) 2008;5(2):288-294
Objective To evaluate the conventional ultrasound (US) and contrast-enhanced ultrasound (CEUS) in diagnosis of splenic trauma including its grading diagnosis. Methods US and CEUS in 42 patients with splenic trauma confirmed by CT and/or operation were performed during 9.2004-10.2007. All the data were compared and analyzed retrospectively. Results Of 42 patients with splenic trauma, 28 cases were detected and 14 cases were missed on US examination, whereas, 40 cases were detected and only 2 mild cases were missed on CEUS examination. The detection rate of lesions with CEUS was significantly higher than that of US (P<0.001) . Ten cases in grading the injury were underestimated by US, however, none of them were underestimated by CEUS. CEUS had good concordance with CT and/or operation in grading diagnosis of 42 cases except two mild cases. Conclusions CEUS has very good concordance with CT and/or operation in detecting injury and grading the splenic trauma compared with conventional US. CEUS as a new imaging technology has made great advance of ultrasoongraphy in evaluation of splenic trauma including injury grading,and it is very useful in clinical application.
9.Non-invasive prenatal diagnosis of trisomy 21 by dosage ratio of fetal chromosome-specific epigenetic markers in maternal plasma.
Ming, ZHANG ; Tao, LI ; Jingyi, CHEN ; Li, LI ; Chun, ZHOU ; Yan, WANG ; Wenhui, LIU ; Yuanzhen, ZHANG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2011;31(5):687-92
This study examined the methylation difference in AIRE and RASSF1A between maternal and placental DNA, and the implication of this difference in the identification of free fetal DNA in maternal plasma and in prenatal diagnosis of trisomy 21. Maternal plasma samples were collected from 388 singleton pregnancies, and placental or chorionic villus tissues from 112 of them. Methylation-specific PCR (MSP) and methylation-sensitive restriction enzyme digestion followed by fluorescent quantitative PCR (MSRE + PCR) were employed to detect the maternal-fetal methylation difference in AIRE and RASSF1A. Diagnosis of trisomy 21 was established according to the ratio of fetal-specific AIRE to RASSF1A in maternal plasma. Both methods confirmed that AIRE and RASSF1A were hypomethylated in maternal blood cells but hypermethylated in placental or chorionic villus tissues. Moreover, the differential methylation for each locus could be seen during the whole pregnant period. The positive rates of fetal AIRE and RASSF1A in maternal plasma were found to be 78.1% and 82.1% by MSP and 94.8% and 96.9% by MSRE + PCR. MSRE + PCR was superior to MSP in the identification of fetal-specific hypermethylated sequences (P<0.05). Based on the data from 266 euploidy pregnancies, the 95% reference interval of the fetal AIRE/RASSF1A ratio in maternal plasma was 0.33-1.77, which was taken as the reference value for determining the numbers of fetal chromosome 21 in 102 pregnancies. The accuracy rate in 98 euploidy pregnancies was 96.9% (95/98). Three of the four trisomy 21 pregnancies were confirmed with this method. It was concluded that hypermethylated AIRE and RASSF1A may serve as fetal-specific markers for the identification of fetal DNA in maternal plasma and may be used for noninvasive prenatal diagnosis of trisomy 21.
10.Transient folate deprivation in combination with small-molecule compounds facilitates the generation of somatic cell-derived pluripotent stem cells in mice.
Wen-tao, HU ; Qiu-yue, YAN ; Yu, FANG ; Zhan-dong, QIU ; Su-ming, ZHANG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2014;34(2):151-6
Induced pluripotent stem cells (iPSCs) can be propagated indefinitely, while maintaining the capacity to differentiate into all cell types in the body except for the extra-embryonic tissues. This iPSC technology not only represents a new way to use individual-specific stem cells for regenerative medicine but also constitutes a novel method to obtain large numbers of disease-specific cells for biomedical research. However, the low efficiency of reprogramming and genomic integration of oncogenes and viral vectors limit the potential application of iPSCs. Chemical-induced reprogramming offers a novel approach to generating iPSCs. In this study, a new combination of small-molecule compounds (SMs) (sodium butyrate, A-83-01, CHIR99021, Y-27632) under conditions of transient folate deprivation was used to generate iPSC. It was found that transient folate deprivation combined with SMs was sufficient to permit reprogramming from mouse embryonic fibroblasts (MEFs) in the presence of transcription factors, Oct4 and Klf4, within 25 days, replacing Sox2 and c-Myc, and accelerated the generation of mouse iPSCs. The resulting cell lines resembled mouse embryonic stem (ES) cells with respect to proliferation rate, morphology, pluripotency-associated markers and gene expressions. Deprivation of folic acid, combined with treating MEFs with SMs, can improve the inducing efficiency of iPSCs and reduce their carcinogenicity and the use of exogenous reprogramming factors.