1.T-2 toxin inhibits mitochondrial function of differentiated murine embryonic stem cells
Haiqin FANG ; Lizhong LI ; Zengming ZHAO ; Jun HE ; Jun ZHAO ; Rong YANG ; Xue GENG ; Shuangqing PENG
Chinese Journal of Pharmacology and Toxicology 2014;(3):415-420
OBJECTIVE ToexplorethepossiblemechanismoractiontargetsofT-2toxinembryo toxicity by observing the effect of T-2 toxin on mitochondrial function of differentiated murine e mbryonic stemcells(mESCs).METHODS Duringdifferentiationat24,72and120h,ESCswereexposedto T-2 toxin 0.5 μg·L-1 .Meanwhile,mESCs were pre-treated with antioxidant Trolox (200 μmol·L-1 )for 30 min and exposed to T-2 toxin (0.5 μg·L-1 )for 72 h.The mitochondrial ultrasture of differentiated mESCs was observed under a transi mission electrical microscope (TEM).The differentiated ESC mito-chondrial function,including respiratory control ratio (RCR),ATP synthase activity and mitochondrial membranepotential(MMP),wasmeasuredat144hafterdifferentiation.RESULTS Significant decrease of the mitochondrial number,deformation of mitochondrial structure,and lack of complete mito-chodrial crest were observed through TEM in the groups of T-2 toxin exposed for 72 and 1 20 h,respec-tively.Compared with the normal control group,RCR declined by 49.5% and 55.1%,ATP synthase activity decreased by 84.9% and 89.3%,and MMP decreased by 23.2% and 35.2% in T-2 toxin 0.5 μg·L-1 exposure 72 and 1 20 h group,respectively.However,the inhibition of mitochondrial function by T-2 toxin in differentiated mESCs recovered significantly in the presence of the antioxidant Trolox. CONCLUSION T-2toxininducesoxidativestressandinhibitsmESCsmitochondrialfunctionindifferenti-ated mESCs,and ROS-induced mitochondrial malfunction plays an i mportant role in T-2 toxin e mbryonic toxicity mechanis m.
2.Effect of lentivirus-mediated interference with long non-coding RNA LINC00630 expression in vitro on proliferation and migration of bladder cancer cells
Geng HUANG ; Dingwen GUI ; Xiaoying WANG ; Wei PENG ; Yunfei ZHAO ; Jinghua WAN ; Fang XIE
Cancer Research and Clinic 2021;33(4):254-258
Objective:To explore the expression of long non-coding RNA LINC00630 in bladder cancer cell lines, and to explore the effect of interference with its expression in vitro on the proliferation and migration of bladder cancer cells.Methods:Real-time fluorescent quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression of LINC00630 in bladder cancer cell lines 5637, BIU-87, T24, J82 and normal bladder epithelial cell line SV-HUC-1. The bladder cancer cell line with the highest LINC00630 expression was selected for follow-up experiments, then the cell line infected with the control lentivirus was used as the control group, and the cell line infected with the lentivirus that could interfere with the expression of LINC00630 was used as the experimental group. qRT-PCR was used to detect the expression of LINC00630 in the two groups of cells. MTS method and cell scratch test were used to detect the proliferation and migration abilities of cells in the two groups. qRT-PCR was used to detect the expression of neuregulin 1 (NRG1) mRNA in the two groups of cells, and Western blot was used to detect the expressions of NRG1 protein, cell proliferation-related proteins (cyclin D3 and CDK2) and cell migration-related proteins (Vimentin and N-cadherin) in the two groups of cells.Results:Compared with SV-HUC-1 cells (1.05±0.17), the expression of LINC00630 was significantly increased in all bladder cancer cell lines (all P < 0.01), and the expression was highest in J82 cells (relative expression 5.83±0.42). Compared with J82 cells of the control group, the expression of LINC00630 in J82 cells of the experimental group decreased (0.18±0.02 vs. 1.00±0.05, t=14.36, P < 0.01); from day 2 of transfection, the cell proliferation activity of the experimental group was lower than that of the control group (all P < 0.05). The cell scratch closure rate of the experimental group was lower than that of the control group [(27.4±7.1)% vs. (66.0±5.4)%, t = 4.31, P < 0.01]. Therelative expression of NRG1 mRNA in the experimental group was lower than that in the control group (0.34±0.03 vs. 1.07±0.24, t = 2.99, P < 0.05). Compared with the control group, the expressions of NRG1 protein, cell proliferation-related proteins and cell migration-related proteins in the experimental group were reduced. Conclusions:LINC00630 is up-regulated in bladder cancer cell lines, and interference with LINC00630 may inhibit the proliferation and migration of J82 cells by down-regulating the expression of NRG1 gene. LINC00630 may be a new molecular target for the treatment of bladder cancer.
3.The development of a treadmill system.
Zhao-ying GENG ; Hong-xia ZHOU ; Chun-fang ZHANG
Chinese Journal of Medical Instrumentation 2002;26(2):133-137
This paper introduces the designing concept of the ECG treadmill system and discusses the methods of its realization.
Amplifiers, Electronic
;
Computer Simulation
;
Computer Systems
;
Coronary Disease
;
diagnosis
;
Electrocardiography
;
instrumentation
;
methods
;
Equipment Design
;
Exercise Test
;
instrumentation
;
methods
;
Humans
;
Microcomputers
;
Signal Processing, Computer-Assisted
;
Software
4.Establishment of Murine Tumor Cell Line Expressing HPV types 6b and 11 E6/E7 Genes
Kejia ZHAO ; Hao CHENG ; Minli CHEN ; Zhishan DING ; Liyi GENG ; Yongming FANG
Chinese Journal of Dermatology 1994;0(02):-
Objective To construct four expression plasmids, pcDNA3.1-GFP/HPV6bE6, pcDNA3.1-GFP/HPV6bE7, pcDNA3.1-GFP/HPV11E6, pcDNA3.1-GFP/HPV11E7 and their transfected murine cell lines. Methods The Four recombinant expression plasmids comprising HPV6bE6,HPV6bE7,HPV11E6 and HPV11E7 linked with GFP, respectively, were constructed and transfected to B16 cells by lipofectamine kit. Positive clones were selected by G418 and observed by fluorescent microscopy and identified by RT-PCR. Results The four constructed recombinant plasmids were authenticated by restriction enzyme digestion and DNA sequencing. Under the fluorescent microscope, the green fluorescence could be observed in cytoplasm and nucleus of four transfected B16 cell lines. The RNA extracted from positively transfected clones resistant to G418 were analyzed by RT-PCR, which demonstrated the presence of four expected fragments. Conclusions The transfected murine cell lines B16 can express HPV6bE6,HPV6bE7,HPV11E6 and HPV11E7 gene. These transfected cell lines can be further transplanted to mice in order to investigate the biological properties and immunological mechanisms of these genes in vivo.
5.Clinical therapeutic effects and pulmonary function effects of phentolamine combined with Dangshen Bufei Decoction in the treatment of asthmatic disease in infants
Li ZHAO ; Fang LI ; yi Shao GENG
Chinese Traditional Patent Medicine 2017;39(11):2259-2263
AIM To investigate the clinical therapeutic effects and pulmonary function effects of phentolamine combined with Dangshen Bufei Decoction (Astragali Radix,Codonopsis Radix,Atractylodis macrocephalae Rhizoma,etc.) in the treatment of asthmatic disease in infants.METHODS From August 2013 to April 2016,94cases of infants with asthmatic disease in our hospital for diagnosis and treatment were selected and randomly divided into observation group of 47 cases and control group of 47 cases.The control group was given phentolamine treatment,while the observation group was treated with phentolamine combined with Dangshen Bufei Decoction.Two groups were both treated for seven days.RESULTS The total effective rates in the observation group and the control group were 97.9% and 85.1%,respectively,the total effective rate in the observation group was significantly higher than that in the control group (P < 0.05).The total disappearance time of dyspnea,cough,fever and other symptoms in the observation group were significantly faster than those in the control group (P < 0.05).The incidence rate of facial flushing,irritability,bradycardia,nausea,vomiting and other adverse reactions in the observation group was 17.0%,which was 19.1% in the control group,there was no significant difference between the two groups (P >0.05).CD4 + values in the observation group and the control group after the treatment were significantly lower than those before the treatment (P < 0.05),while CD8 + values were significantly higher than those before the treatment (P < 0.05).After the treatment,CD4 + and CD8 + values in the observation group showed significant statistical differences as compared with those in the control group (P < 0.05).FVC values in the observation group and the control group after the treatment were (83.55 ± 15.29)% and (75.20 ± 11.49)%,respectively,which were significantly higher than those before the treatment [(68.24 ± 15.20)% and (69.01 ± 14.03)%,respectively] (P <0.05),and FVC value in the observation group was also significantly higher than that in the control group (P < 0.05).CONCLUSION Phentolamine combined with Dangshen Bufei Decoction has good safety in the treatment of asthmatic disease in infants,which can improve clinical symptoms and immune function,so as to promote the improvement of lung function and therapeutic effects.
6.The capability of high field MRI in demonstrating post-mortem fetal brains at different gestational age
Zhonghe ZHANG ; Shuwei LIU ; Xiangtao LIN ; Gaojun TENG ; Taifei YU ; Fang FANF ; Bin ZHAO ; Fengchao ZANG ; Hequn GENG
Chinese Journal of Radiology 2009;43(11):1131-1134
Objective To study the capability of high field MRI in demonstrating the post-mortem fetal brains at different gestational age(GA).Methods One hundred and eight post-mortem fetal brains of 14-40 weeks GA were evaluated by 3.0 T MRI. Eleven brains of 14 to 27 weeks GA with good 3.0 T MRI images were chosen and scanned by 7.0 T MRI. The developing sulci, layered structures of fetal cerebral cortex and basal nuclei were evaluated on MRI of different Tesla(3.0 T and 7.0 T)and their results analyzed. Results On T_1 WI of 3.0 T MRI, the layered structures of fetal cerebral cortex were present at 14 weeks GA, the sulci were more accurately identified after 16 weeks GA. The basal nuclei were clearly distinguishable after 20 weeks GA. and these structures were better visualized as the GA increased. On T_2WI of 7.0 T MRI, the sulei, layered structures of fetal cerebral cortex and basal nuclei were shown more clearly at the same GA when compared to 3.0 T, especially the sulci at the early developmental stages. Conclusions T_1 WI of 3.0 T MRI could show the developing structures of post-mortem fetal brain well, but the T_2 WI of 7.0 T MRI were comparatively better.
7.Study on the sensitizing potential of shuanghuanglian injection using popliteal lymph node assay in C57BL/6J mice.
Zhao-Hua LIU ; Fang CHENG ; Geng-yin ZHOU
Chinese Journal of Integrated Traditional and Western Medicine 2010;30(1):64-67
OBJECTIVETo investigate the sensitizing potential of Shuanghuanglian Injection (SHL) by comparing the popliteal lymph node (PLN) response in mice induced by SHL and chemicals.
METHODSSixty female C57BL/6J mice were equally and randomly divided into six groups, i.e. the blank control group (A) and five treated groups treated respectively with phenobarbital 1 mg/mouse (B), mercuric chloride ( HgCl2) 50 microg/mouse (C), D-penicillamine 2 mg/mouse (D), and SHL in low (1 mg/mouse) and high (5 mg/mouse) dosages (E and F) via subcutaneous injection into left pad of hind foot. Animals were sacrificed on the 8th day after injection, their bilateral PLNs were isolated and weighed respectively to calculate the PLN mass index (MI). Then the PLNs get from four mice in each group were fixed with 4% paraformaldehyde solution for histopathologic examination; the other six PLNs were prepared into single-cell suspensions to calculate cell index (CI) for comparing the changes of PLN in various groups.
RESULTSMI and CI in Group F reached to > or = 2 and > or = 5 (average) respectively, which was higher than those in Group A (P<0.05). Pathological examination showed that the left PLN in Group F enlarged, with remarkable germinal center and increased high endothelial venules proliferation.
CONCLUSIONSHL could induce significant PLN response in C57BL/6J mice, suggesting it has certain sensitizing potential.
Animals ; Drugs, Chinese Herbal ; adverse effects ; Female ; Hypersensitivity ; pathology ; Local Lymph Node Assay ; Lymph Nodes ; drug effects ; immunology ; pathology ; Mice ; Mice, Inbred C57BL
8.Primary investigation of space mutated tumor cells
Jin-tian TANG ; Qing XIANG ; Mei XU ; Zhihua CHEN ; Hongyan LI ; Qing FANG ; Xuan LIU ; Bo XU ; Chuanying GENG ; Tiande ZHAO ; Weichang CHEN
Chinese Journal of Rehabilitation Theory and Practice 2004;10(11):641-643
ObjectiveTo search for a new method of anti-tumor immunity,using mutated tumor cells by spaceflight.MethodsTumor cells were carried in the Shenzhou 4 and the recoverable satellite No.18.After 7 days and 18 days spaceflight respectively,the effects of spaceflight were investigated primarily.Results2382 and 1 strains of mutated tumor cells from the airship No.4 and the recoverable satellite No.18 had been obtained respectively. Comparing with the control group, the growth rate of mutated cells decreased, moreover, the secretion of cytokines also changed.ConclusionSpaceflight may affect physiological characteristics of tumor cells, and that, there was a negative correlation between the ratio of surviving cells and carrying time.
9.Relation between glycosylated hemoglobin and acute cerebral infarction
Qing-Bo GE ; Qiao-Yun LUO ; Jing LI ; Zhao-Geng FANG ; Zhen-Zhen WANG ; Zhi-Hong LIU
Chinese Journal of Neuromedicine 2010;09(7):715-718
Objective To investigate the differences of blood glycosylated hemoglobin (HbA1c) levels between the patients with acute cerebral infarction and healthy controls, and explore the relation between HbA1c level and both neurological deficits scores and imaging of lesions in number. Methods One hundred and eighty-six patients with acute cerebral infarction within 1 week were performed neurological deficits scales after the admission; the HbA1c level was measured within 24 h admission; brain MRI scan was performed on these patients 48 h after onset or stabilization. Glucose tolerance test was taken at the rehabilitation of infarction (except for having a clear history of diabetes before). At the same time, 160 healthy controls were checked on the level of HbA1c and taken the glucose tolerance test. The differences of blood HbA1c levels between the patients with acute cerebral infarction and healthy controls were investigated; and the relation between HbA1c level and both neurological deficits scores and imaging of lesions in number was explored. Results The HbA1c level in patients with acute cerebral infarction (6.982%±1.803%) was significantly higher than that in the controls (5.128%±0.592%, P<0.05). The level of HbA1c in patients with cerebral infarction and the neurological deficits scores were positively correlated (r=0.760, P<0.05). The level of HbA1c in patients with 2 lesions (6.635%±0.427%) was obviously higher than that in patients with 1 lesion (5.803%±0.307%, P<0.05); The level of HbA1c in patients with 3 or more lesions (8.571%±0.519%) was obviously higher than that in patients with 1 or 2 lesions (P<0.05). Conclusion Diabetes is a major risk factor for cerebral infarction. High HbA1c level might cause a series of cerebrovascular diseases, thus it is an important factor in the happening of cerebral infarction and HbA1c level is an important indicator of the early assessment of the severity of the diseases. The incidence of cerebral infarction can be decreased by controlling hyperglycaemia, lowering the HbA1c levels.
10.Inhibition of 6A8 alpha-manosidase expression induces decrease of adhesion to laminin and reduction of lamellipodia of human nasopharyngeal carcinoma cell CNE-2L2.
Wei YUE ; Fang-tao ZHAO ; Geng-xian SHI ; Yin LIU ; Li-ping ZHU
Acta Academiae Medicinae Sinicae 2003;25(3):316-319
OBJECTIVETo investigate the inhibitory effect of 6A8 alpha-manosidase expression on the adhesiveness of CNE-2L2 cells to laminin and the lamellipodia on cell surface.
METHODS6A8 alpha-manosidase expression was detected by Western blotting. For assaying the adhesion of cells to laminin, cells were incubated in laminin-coated plate at 37 degrees C for 1 h, the adhered cells were stained with crystal purple dissolved in 0.1 mol/L Sodium Citrate/50% ethanol. Absorbance 540 nm was measured. Adhesion rate (R) was calculated according to formula R = AT/A100 x 100%. Here A100 represents 100% adhesion. lamellipodia on cell surface was observed upon a scanning electron microscopy.
RESULTSThe adhesion rate of two clones (AS1 and AS2) with inhibition of 6A8 alpha-manosidase expression to laminin was 0.447 +/- 0.096 and 0.533 +/- 0.065 respectively. The adhesion rate of three controls with normal expression of 6A8 alpha-manosidase to laminin was 0.78 +/- 0.035, 0.7 +/- 0.05 and 0.80 +/- 0.04 respectively. The difference was significant (P < 0.01). CNE-2L2 cells with normal expression of 6A8 alpha-manosidase was rich in lamellipodia on their surface. Lamellipodia nearly disappeared on the cells with inhibition of 6A8 alpha-manosidase expression.
CONCLUSIONSInhibition of 6A8 alpha-manosidase expression results in decrease of adhesion to laminin and reduction of lamellipodia of human nasopharyngeal carcinoma cell CNE-2L2.
Cell Adhesion ; drug effects ; Humans ; Laminin ; physiology ; Nasopharyngeal Neoplasms ; pathology ; Neoplasm Metastasis ; Neoplasm Proteins ; genetics ; physiology ; Pseudopodia ; physiology ; Tumor Cells, Cultured ; alpha-Mannosidase ; biosynthesis ; genetics