1.Low dose hyper-radiosensitivity in human lung cancer cell line A549 and its possible mechanisms
Dan TAO ; Jing CHENG ; Gang WU ; Hongge WU ; Jun XUE
Chinese Journal of Radiological Medicine and Protection 2009;29(2):147-151
Objective To study the low dose hyper-radiosensitivity in human lung cancer cell line A549,and its possible mechanisms.Methods Exponentially growing A549 cells were irradiated with 60Co γ-rays at doses of 0-2 Gy.Together with flow cytometry for precise cell sorting,cell survival fraction was measured by mean of conventional colony-formation assay.ATM1981 Ser-P protein expression was examined by Western blot.Apoptosis was identified by Hoechst 33258 fluorescent staining,and Annexin V-FITC and propidium iodide staining flow cytometry.Cell cycle distribution was observed by flow cytometry.Results There was an excessive cell killing per unit dose when the doses were below about 0.3 Gy,and the cells exhibited more resistant response at the doses between 0.3 and 0.5 Gy,the cell survival fraction was decreased as the doses over 0.5 Gy.The expression of ATM1981Ser-P protein was first observed at 0.2 Gy,followed by an increase over 0.2 Gy,and reached the peak at 0.5 Gy(compared with 0.2 Gy group,t=7.96,P<0.05),with no further increase as the doses at 1.0 and 2.0 Gy(t=0.69,0.55,P>0.05).24 hours after irradiation,part cells presented the characteristic morpholos4cal change of apoptosis,and the apoptosis curve was coincident with the dose-survival curve.Compared with the control group,the cell cycle had no change post-irradiation to 0.1 and 0.2 Gy.G2/M phase arrest was manifested at 6 and 12 hours post-irradiation to 0.3,0.4 and 0.5 Gy(t=2.87,2.88,4.92 and 3.70,3.12,8.11,P<0.05),and the ratio of G2/M phase was decreased at 24 hours post-irradiation(t=3.87,4.77,3.01,P<0.05).Conclusions A549 cells displays the phenomenon of hyper-radiosensitivity(HRS)/induced radioresistance(IRR).The model of cell death induced by low dose irradiation is mainly apoptosis.The activity of ATM and cell cycle change might play an important role in HRS/IRR.
2.Double lethal effects of fusion gene of wild-type p53 and JunB on hepatocellular carcinoma cells.
Cheng, GUO ; Qingguang, LIU ; Lei, ZHANG ; Xue, YANG ; Tao, SONG ; Yingmin, YAO
Journal of Huazhong University of Science and Technology (Medical Sciences) 2012;32(5):663-8
This study explored the double lethal effects of pEGFP-C1-wtp53/junB fusion gene on hepatocellular carcinoma (HCC) cells. wtp53/junB fusion gene was constructed and transformed into HepG2 cell line. Expression of KAI1 was detected by quantitative real-time PCR and Western blotting, cells apoptosis rate was detected by flow cytometry, proliferation of cells was detected byMTT chromometry, cell transmigration was detected by using transwell systems. The results showed that after transformation with pEGFP-C1-wtp53/JunB, the expression level of KAI1 protein was up-regulated, being 8.13 times the blank control group in HepG2 cells and significantly higher than 2.87 times which transformed with pEGFP-C1-JunB, 3.11 times which transformed with pEGFP-C1-wtp53 (P<0.001). Apoptosis rate of HepG2 cells transformed with pEGFP-C1-wtp53/JunB was significantly higher than that of other groups (P<0.001), and invasive ability of HepG2 cells transformed with pEGFP-C1-wtp53/JunB was significantly lower than other groups(P<0.001). It was concluded that the fusion gene of wtp53 and JunB could not only inhibit the growth of hepatoma cells and promote tumor cell apoptosis, but also suppress the invasive ability of tumor cells by up-regulating the expression of KAI1.
3.Effects of HSP70 antisense oligonucleotide on the proliferation and apoptosis of human hepatocellular carcinoma cells.
Xue, YANG ; Haibin, HE ; Wei, YANG ; Tao, SONG ; Cheng, GUO ; Xin, ZHENG ; Qingguang, LIU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2010;30(3):337-43
The study investigated the effects of heat shock protein 70 (HSP70) antisense oligonucleotide (ASODN) on the proliferation and apoptosis of a human hepatocellular carcinoma cell line (SMMC-7721 cells) in vitro. HSP70 oligonucleotide was transfected into SMMC-7721 cells by the mediation of Sofast transfection reagent. Inhibition rate of SMMC-7721 cells was determined by using MTT method. Apoptosis rate and cell cycle distribution were measured by flow cytometry. Immunocytochemistry staining was used to observe the expression of HSP70, Bcl-2 and Bax. The results showed that HSP70 ASODN at various concentrations could significantly inhibit the growth of SMMC-7721 cells, and the inhibition effect peaked 48 h after transfection with 400-nmol/L HSP70 ASODN. Cytometric analysis showed the apoptotic rate was increased in a dose- and time-dependent manner in the HSP70 ASODN-treated cells. The percentage of cells in the G(2)/M and S phases was significantly decreased and that in the G(0)/G(1) phase increased as the HSP70 ASODN concentration was elevated and the exposure time prolonged. Immunocytochemistry showed that treatment of SMMC-7721 cells with HSP70 ASODN resulted in decreased expressions of HSP70 and Bcl-2 proteins, and an increased expression of Bax protein. It was concluded that the HSP70 ASODN can inhibit the growth of the SMMC-7721 cells and increase cell apoptosis by down-regulating the expression of HSP70. HSP70 ASODN holds promise for the treatment of hepatocellular carcinoma.
4.Construction of eukaryotic expression vector of wtp53/junB fusion gene
Cheng GUO ; Lei ZHANG ; Qingguang LIU ; Tao SONG ; Xue YANG ; Xin ZHENG ; Yingmin YAO
Journal of Xi'an Jiaotong University(Medical Sciences) 2010;31(1):41-46
Objective To construct wtp53/junB fusion gene and its eukaryotic expression vector in order to provide the basis for further application of polygene union therapy in hepatocellular carcinoma. Methods Polymerase chain reaction (PCR), reverse transcription-PCR (RT-PCR) and gene recombination techniques were used to construct the eukaryotic vector of pEGFP-C1-wtp53/junB fusion gene, which carries the enhanced green fluorescent protein (EGFP). The transfection of pEGFP-C1-wtp53/junB in hepatoma HepG2 cells was detected by the location of green fluorescence. Results The DNA sequence of wtp53/junB fusion gene was successfully cloned into the pEGFP-C1 plasmid and the sequence was the same as what we expected. Green fluorescence located on cell nucleus proved that pEGFP-C1-wtp53/junB was transfected into HepG2 cell line successfully. Conclusion We successfully constructed the eukaryotic vector of pEGFP-C1-wtp53/junB fusion gene, which carries the EGFP, and transfects it into human hepatoma cell nucleus. It may lay the basis for studying the synergetic effect of wtp53 and junB in hepatocellular carcinoma.
5.Effects of Tangshenkang on MMP-9 and TIMP-1 of Human Renal Tubular Epithelial Cell HK-2 Induced by High Glucose
Qingchun CHAI ; Lixia YANG ; Jianjun XUE ; Tao CHENG ; Dinghua ZHANG ; Tonghua LIU
Chinese Journal of Information on Traditional Chinese Medicine 2014;(10):58-60
Objective To explore the mechanism of Tangshenkang in the prevention and treatment of diabetic nephropathy. Methods HK-2 cells were cultured in vitro and divided into control group, high glucose group (30 mmol/L D-glucose), control group (30 mmol/L D-glucose+10% animal serum), and Tangshenkang drug-containing serum therapy groups (30 mmol/L D-glucose+5%low concentration Tangshenkang, 30 mmol/L D-glucose+10%middle concentration Tangshenkang, 30 mmol/L D-glucose+20% high concentration Tangshenkang). After 24 h and 48 h treatment, MMP-9 and TIMP-1 in cell cultural supernatant were observed by ELISA. Results MMP-9 of HK-2 cultured with high glucose was much decreased and TIMP-1 increased significantly than the control group, with statistical significance (P<0.05). TIMP-1 significantly decreased and MMP-9 increased in HK-2 cultured with high glucose plus Tangshenkang compared with those only induced by high glucose, with statistical significance (P<0.05). Conclusion Tangshenkang could regulate the secretion of fibrosis cell factor of HK-2 cell induced by high glucose, which may be one of the mechanisms in its treatment of diabetic nephropathy.
6.Early and late outcome of acute type A aortic dissection in dialysis dependent patients
Lichong LU ; Min GE ; Tao CHEN ; Cheng CHEN ; Zhigang WANG ; Jiaxin YE ; Dongjin WANG ; Yunxing XUE
Chinese Journal of Thoracic and Cardiovascular Surgery 2021;37(4):216-219
Objective:To summarize acute type A aortic dissection(ATAAD) is relatively uncommon in dialysis patients, and repair outcomes are not fully understood.Methods:Between January 2014 and March 2020, 20 patients with ATAAD required dialysis for preoperative end-stage renal disease(ESRD) were treated by our group. There were 11 male and 9 female patients at mean age of(47.8±11.3) years. The mean duration of dialysis therapy in the total 20 patients before the onset of ATAAD was(4.5±3.9 )years, with 90%(18 cases) of these patients undergoing hemodialysis(rather than peritoneal dialysis). 17 patients were treated emergency surgically, surgical operation were performed under deep hypothermic circulatory arrest and perfused the cerebral selective cerebral perfusion, 5 cases with ascending aorta + arch fenestrated stent, 5 cases with ascending aorta+ hemi-arch replacement(2 cases with stent elephant trunk), 4 cases with ascending aorta+ arch replacement+ stent elephant trunk(1 case with coronary artery bypass grafting for left anterior descending coronary artery), 2 cases with aortic valvuloplasty + ascending aorta+ hemi-arch replacement, 1 case with Bentall+ arch fenestrated stent.Results:2 patients were died from aortic ruptured before operation, 1 patient treated medically was alive three months after admission. Cross-clamp, cardiopulmonary bypass, and circulatory arrest times of all the surgical patients were(233.8±84.4) min, (155.5±63.6)min and(28.2±10.8)min, respectively. The following complications occurred postoperative: 3 cases died in the hospital, 1 case of tracheotomy, 2 cases of cerebral infarction, 1 case of cerebral hemorrhage, 1 case of transient paraplegia, and 1 case of surgical site infection. After a mean follow-up of(11.6±14.5) months(rang, 3-61 months). the overall survival rate at 1 year and 5 years was 53% and 27% respectively.Conclusion:Dialysis patients with ATAAD should be operated on urgently and medical treatment carries high risks of aortic rupture, although in-hospital mortality is acceptable, long-term mortality is poor.
7.Anti-infection effect of phage and its clinical application
Xue XIONG ; Tao CHEN ; Yajun LIU ; Liting CHENG ; Xingmei WANG ; Huaping LIANG
Chinese Critical Care Medicine 2021;33(4):497-499
In recent years, the problem of bacterial resistance has become more and more serious, which has brought troubles to global public health and medical care. The time and money required to develop new antibiotics is even greater than before. Bacteriophage is a kind of virus that can specifically infect bacteria, fungi, actinomycetes and other microorganisms. Relying on host bacteria to replicate in large numbers, rich species, low research and development cost, the value of anti-infection therapy is very considerable. It is a new generation of biological antimicrobial agents with great potential. This paper briefly describes the sterilization mechanism, progress of research on anti-infection aspect and clinical application of phage, in order to provide reference for phage anti-infection treatment and clinical application.
8.Mental health status of 990 rural community elderly and its associated factors in Chongqing
Daijiang ZHANG ; Xiaolin TAN ; Xue CHENG ; Bo WANG ; Jianchu ZHOU ; Guoxing CAO ; Liming CHEN ; Tao LIU ; Yuan LI
Chongqing Medicine 2014;(7):775-778
Objective To explore mental health status of rural elderly and itassociated factors of mental health in Chongqing . Methods Totally 990 rural community residents aged 60 years or more were selected by multi-stage stratified random sampling and investigate with self-designed screening inventory to pick out suspect psychosis ,then diagnose according to DSM-Ⅳ and CCMD-3 . For no psychosis ,investigated with symptom self-assessment scale(SCL-90) ,newfoundland happiness scale ,social support rating scale ,eysenck personality questionnaire short form scale (China) .Results (1) In 990 the elderly ,the screening positive rate was 24 .04% (238/990) ,prevalence rate was 19 .43% (193/990) .Diagnosis were mental disorders caused by acohol(11 .4% ) ,mental dis-orders due to physical dysfuction (5 .96% ) ,mood disorders and other psychiatric disorders (2 .12% ) .(2)17 .17% (124/722) rural elderly had psychological problems in 799 non-mental health elderty ,in which the screening positive rate and total scores of SCL-90 in family group were significant higher than the institute group screening positive rate (screening positive 20 .24% vs .3 .73% ,χ2 =20 .90 ,P=0 .000 ;total scores(109 .14 ± 11 .77)vs .(100 .59 ± 7 .87) ,t= -8 .01 ,P= 0 .000) .(3)According to the total score of SCL-90 ,those female ,without spouse ,negative life events ,live alone ,with chronic disease ,with little income had worse mental health(P<0 .05) .(4)Total score of SCL-90 in rural elderly was significant negatively correlated with total score of happiness ,posi-tive emotions ,positive experience ,all dimensions of social support and internal and external .the score was significant positively cor-related with negative emotions ,negative experience ,neuroticism ,psychoticism ,the differences were statistically significant ( P<0 .05) .(5)Negative emotional ,mental quality ,chronic diseases ,endowment way ,marital status ,whether to live alone ,total score of social support ,neurotic entered the regression equation ,determination coefficient R2 =0 .346 .Conclusion More than 1/3 of the ru-ral elderly with mental health problems ,the pension way was the most obvious impact .prevention and treatment of alcohol-induced mental disorders ,changes in pension mode was still the focus of rural elderly mental health work .
9.Autologus bone marrow-derived mesenchymal stem cells intracoronary delivery after acute myocardial infarction in miniature pig.
Cheng-qian YIN ; Ji-lin CHEN ; Yun-fang WANG ; Feng CAO ; Rui CHENG ; Xue-tao PEI
Acta Academiae Medicinae Sinicae 2005;27(6):696-699
OBJECTIVETo evaluate the possibility that using intracoronary delivery of autologus bone marrow-derived mesenchymal stem cells (MSCs) to improve the cardiac function after acute myocardial infarction (AMI) in miniature pig.
METHODSMSCs were cultured in Dulbecco's modified Eagle's medium-F12 (DMEM/F12) medium. AMI model was made by blocking the blood stream of the first diagonal branch in miniature pig, and released the branch after 90 minutes. After 10-14 days, (4-6) x 10(7) culture-expanded autologus 4', 6-diamidino-2-phenylindole (DAPI)-labelled MSCs were transplanted into each host heart's AMI area through intracoronary way. Ultrasonic cardiography (UCG) was performed to observe the left ventricular function at 3 months after transplantation. The cellular transplanted hearts were harvested and investigated by immunohistochemical analysis.
RESULTSLeft ventricular function of the MSCs group was improved significantly 3 months later compared with the control group [(54.65 +/- 3.39) vs (43.98 +/- 4.21)%, (P < 0.01)]. Exogenous MSCs survived and site-differentiated into cardiomyocytes in infracted hearts.
CONCLUSIONMSCs can play a benificial role to repair damaged heart. Heart function can be improved after MSCs transplantation in porcine myocardial infarction model.
Animals ; Female ; Male ; Mesenchymal Stem Cell Transplantation ; methods ; Myocardial Infarction ; pathology ; physiopathology ; therapy ; Swine ; Swine, Miniature ; Transplantation, Autologous ; Treatment Outcome
10.Seroepidemiology investigation of high-risk population with plague foci in Ningxia in 2007 and 2008
Ying-xu, QIN ; Jian-wei, GAO ; Tao, TIAN ; Xue-ping, MA ; Shi-tang, LU ; Sheng-long, ZHANG ; Yong, ZHU ; Xue-dong, SUN ; Xiang-yang, XUE ; Wei-cheng, FU ; Hong-ya, WANG
Chinese Journal of Endemiology 2010;29(4):443-445
Objective To understand the level and distribution of antibody F1 against plague in population of Ningxia natural plague foci in 2007 and 2008. Methods Seven hundred and eighteen blood samples were collected in five major cities and counties of natural plague foci, and 475 blood samples were collected in nonplague area as control group. Conventional indirect hemagglutination, colloidal gold test, and enzyme-linked immunoassay were employed to test the antibody. If the result was tested positive by more than two methods used then the result was defined as positive. Antibody titer that did not reach the positive standard was defined as suspected samples. Results A total of 718 serum samples were tested, the results showed that 9 samples were positive (antibody titer was 1:16 - 1:64), the positive rate was 1.25%(9/718), suspected samples was 28, the detection rate was 3.90%(28/718). Four hundred and seventy-five serum samples in the non-plague area were all negative by the three methods. There was a significant difference of antibody F1 positive rate between residents in historical epidemic area and history nonepidemic area(χ2 = 4.44, P< 0.05). There was no statistical significance of the positive rate[1.25%(9/718), 1.25%(9/718),2.51%(18/718)]among the three methods used(χ2 = 1.91, P> 0.05). Conclusion There still exists a certain proportion of Fl antibody positive people in Ningxia natural plague foci, and these people are distributed in areas where several animal plague prevalent in recent years.