1. Over-expression of transducer of ErbB2, 1 inhibits migration and invasion of human lung cancer cell line A549 in vitro
Tumor 2012;32(2):85-91
Objective: To investigate the effect of over-expression of transducer of ErbB2, 1 (TOB1) on migration and invasion of human lung adenocarcinoma cell line A549. Methods: The expression levels of TOB1, phosphatase and tensin homolog deleted on chromosome ten (PTEN), breast cancer metastasis suppressor 1 (BRMS1), ras homolog gene family, member C (RHOC) and non-metastatic cells 1 (NME1) mRNAs in human bronchial epithelial (HBE) cells and ten types of human lung cancer cells were detected by RT-PCR. The recombinant plasmid pcDNA3.0-TOB1 and the empty vector pcDNA3.0 were transfected into A549 cells, respectively. The A549 cells over-expressing TOB1 protein (A549/TOB1 cells) and the A549/PC3 cells (as the control cells) were established. The abilities of migration and invasion of A549/TOB1 cells were determined by wound healing and Transwell assay. The expression levels of PTEN, BRMS1, RHOC and NME1 mRNAs were detected by RT-PCR, and the expression levels of α-N-catenin, β-catenin, γ-catenin, matrix metalloproteinase 2 (MMP2) and MMP9 were detected by Western blotting. Results: The expression levels of TOB1 mRNA in ten types of lung cancer cells were lower than that in HBE cells (P <0.05). The expression level of TOB1 mRNA was positively correlated with the expression levels of PTEN, BRMS1 and NME1 mRNAs (P <0.05). The abilities of migration and invasion of the A549/TOB1 cells were obviously decreased as compared with those of the A549 cells (P <0.05). The expression levels of PTEN, BRMS1 and NME1 mRNAs were higher whereas the expression level of RHOC mRNA was lower in the A549/TOB1 cells as compared with those in the A549 cells (P <0.05). The expression levels of α-N-catenin, β-catenin, γ-catenin, MMP2 and MMP9 proteins in the A549/TOB1 cells were lower than those in the A549 cells (P <0.05). Conclusion: Exogenous TOB1 over-expression can inhibit the abilities of migration and invasion of A549 human lung cancer cells in vitro . This effect may be associated with the downregulation of α-N-catenin, β-cateninm γ-catenin, MMP2 and MMP9 expressions, and which was induced through the regulation of PTEN, BRMS1, RHOC and NME1 expressions by TOB1. Copyright © 2012 by TUMOR.
2.Effects of nicorandil, glutamine, metoprolol used in single and combination on antiapoptosis ability of cardiocytes and expression of HSP70 against myocardial ischemia/reperfusion injury in rats
Wenhua CHEN ; Chang SUN ; Ying ZHANG ; Yanna WU ; Ke WEN ; Yi KANG ; Jianshi LOU
Chinese Pharmacological Bulletin 2014;(9):1242-1246
Aim To investigate the protective effects of nicorandil, glutamine and metoprolol used in single and combination on the antiapoptosis ability of cardio-cytes and the expression of HSP70 after myocardial is-chemia/reperfusion injury in rats. Methods Male Wistar rats were divided randomly into seven groups:( 1 ) Sham group:in which the coronary artery was not roped;(2) I/R group:in which only 30 min ischemia and 120 min reperfusion of LAD was executed; ( 3 ) Nicorandil group; ( 4 ) Glutamine group; ( 5 ) Meto-prolol group; ( 6 ) Nic + Glu + Met ( NGM ) group;(7) low dose of Nic+Glu+Met ( NGML) group. In the pharmacological precondition groups, correspond-ing drugs were administered 30 min before I/R proto-col, and the initial drug dose in each group was 1 mg ·kg-1 . Myocardial apoptotic rates were measured with TUNEL ( terminal dexynucleotidyl transferase-mediated dUTP nick end labeling) , and the expression of apop-tosis related proteins-Bcl-2/Bax and protective pro-tein-HSP70 was detected by Western blot. ResultsIn I/R group, the apoptotic rate in ischemic region was very high ( 36.9% ± 10.3%) , and Bcl-2/Bax was very low ( 0.14 ±0.08 ) . Compared with I/R group, the apoptotic rate of pharmacological precondition groups was decreased significantly ( P <0.01 ) , and the ratio of Bcl-2 to Bax was raised ( P<0.01 ) . Com-pared with Sham group, the expression of HSP70 in-creased significantly ( P<0.01 ) in I/R group. Com-pared with IR group, the expression of HSP70 of phar-macological precondition groups was elevated ( P <0.01 ) , and the expression in NGM group was higher than in the single drug group ( P<0.01 ) . Conclusion Compared with single therapy after I/R in rat, the combination therapy of nicorandil, glutamine, meto-prolol can decrease the apoptotic rate, and the expres-sion of apoptosis related proteins is developed. In the mean time, the expression of protective protein HSP70 is elevated.
3.Leptin receptor of the hind brain nuclei is involved in the conditioned taste preference of rats.
Cai-xia LIN ; Shao-yun ZHANG ; Ke CHEN ; Xiao LUO ; Bo SUN ; Yu-ming KANG ; Jian-qun YAN
Chinese Journal of Applied Physiology 2015;31(6):549-555
OBJECTIVEConditioned taste preference (CTP) is a taste learning reflex by which an animal learns to prefer a substance which tastes not well and has been studied with much interest in recent years. However, the neural substrates of CTP are less known. This study aimed to determine the possible neural path- ways of CTP and whether serum leptin level and the leptin receptor (OB-Rb) in the hind brain are involved following CTP formation.
METHODSWe established CTP of quinine in rats with a 2-bottle preference test. The serum leptin concentrations were detected, the expression of c-fos in the rat brain was tested to determine the nuclei in relation with establishment of CTR Finally, the OB-Rb mRNA expression was examined by RT-qPCR assay in parabrachial nucleus (PBN) and the nucleus of the solitary tract (NST) of the hind brain.
RESULTSCompared with control group, the level of serum leptin was higher in the CTP group (4.58 ± 0.52 vs 1.67 ± 0.25 µg/L, P < 0.01); increased c-fos positive cells were found in the anterior hypothalamus (AH, 221.75 ± 4.96 vs. 178.50 ± 6.63 cells/mm², P < 0.05), the basal lateral amygdala (BLA, 70.75 ± 6.17 vs 56.50 ± 3.62 cells/ mm², P < 0.05) and the nucleus of the solitary tract (NST, 41.25 ± 1.32 vs 32.50 ± 1.02 cells/mm², P < 0.05). But in ventromedial nucleus of the hypothalamus (VMH, 20.75 ± 2.73 vs 38.5 ± 1.54 per 1 mm², P < 005), PBN (21.50 ± 2.24 vs 36.25 ± 1.49 cells/mm², P < 0.05) and the central nucleus of the amygdala (CeA, 22.25 ± 1.53 vs 35.50 ± 2.11 cells/mm², P < 0.05), the number of c-fos positive cells was decreased in the CTP group. In addition, we found OB-Rb mRNA expression in PBN of CTP group rats was higher than that of control group (0.95 ± 0.055 vs 0.57 ± 0.034, P < 0.05), while there was no significant difference of OB-Rb mRNA expression in NST between the two groups.
CONCLUSIONNuclei AH, BLA, NST, VMH, PBN and CeA participate in the formation of CTP. Leptin and its receptor in PBN may be involved in the formation and maintenance of CTP.
Animals ; Conditioning (Psychology) ; Leptin ; blood ; Rats ; Receptors, Leptin ; physiology ; Rhombencephalon ; physiology ; Taste ; physiology
4.Study on enterovirus infection in family close contacts of patients with hand, foot and mouth disease and the contamination of environment
Limei SUN ; Jinyan LIN ; Hanri ZENG ; Min KANG ; Huanying ZHENG ; Aiping DENG ; Jianfen HE ; Changwen KE ; Yonghui ZHANG
Chinese Journal of Infectious Diseases 2012;30(6):339-342
Objective To understand the enterovirus infection in family close contacts of patients with hand,foot and mouth disease (HFMD) and the contamination of environment.Methods Forty-one HFMD cases confirmed by laboratory from web-based surveillance system during July to August 2010 in Guangdong Province were selected.All members of the cases′ family were investigated by collecting their information on demography,habit of domestic hygiene and hygiene status in household.The stool samples of all members and the smear samples from the surface of family belongings from 16 families were collected and the enterovirus was detected by real time quantitative polymerase chain reaction.The data were analyzed by chi square teat and t test.ResultsForty-one HFMD cases′ families and 135 close contacts were included in this survey.The infection rate of the enterovirus was 39.2% (53/135) in all close contacts.Of all the investigated families,the infectionrate was 58.5% (24/41) in family with one or more close contacts and 9.8% (4/41) in family with all close contacts.The differences of infection rates of enterovirus among the members of parents (32.5%,25/77),grandparents/aunts/ uncles (43.3%,13/30) and cousins (53.6%,15/28) didn′t show statistical significance (χ2 =4.07,P=0.131).The infection rate of enterovirus in close contacts from family with more than 5 members was higher than that from family with 4 or less members (OR=1.4,95%CI 1.1-1.9).Among 135 close contacts,27.4% (37/135) were infected with the same types of entervirus as that of HFMD case in the family and 11.9% (16/135) were infected with the different virus types.In 33 family belongings samples from 16 families,the positive rate of enterovirus detection was 6.1% (2/33).Between 17 families with enterovirus testing negative and 23 families with enterovirus testing positive in close contacts,there were no statistical differences of the family hygiene status,hand-washing of babysitter,disinfection of tableware and drinking,sharing towels,airing bedding articles and toy cleaning (P>0.05).ConclusionsThe infection rate of enterovirus in close contacts of HFMD cases is high and the enterovirus contamination exists in case family environment.Management of close contacts of HFMD cases and disinfection of the family environment are important in HFMD controls.
5.Study on sini decoction in treatment of intestinal ischemia-reperfusion injury in rats: mechanism relating to oxygen radical and bcl-2 protein.
Ke-Xuan LIU ; Wei-Kang WU ; Wei HE ; Hui-Lan SUN
China Journal of Chinese Materia Medica 2006;31(4):329-348
OBJECTIVETo investigate the protective effect of Sini decoction (SND) on intestinal mucosa in rats with intestinal ischemia reperfusion (I/R) and the mechanism relating to oxygen radical and Bcl-2 protein expression.
METHODThirty-two SD rats of both sexes were randomly divided into 4 equal groups: (1) control group in which sham operation was performed; (2) model group in which intestinal I/R was produced by clamping super mesenteric artery(SMA) for 1 hour and declamping SMA for 3 hours; (3) SND low dose group in which SND(3 g x kg(-1) x d(-1)) was given via stomach tube for 3 days before operation; (4) SND high dose group in which SND(6 g x kg(-1) x d(1)) was given via stomach tube, for 3 days before operation. A strip of small intestine was taken from distal end of ileum for light microscopic examination, Chiu's score and the detection of intestinal water content (IWC). Apoptosis of intestinal mucosa cell was examined by TUNEL method. Superoxide dismutase (SOD) activity, malondisldehyde (MDA) concentration of intestinal mucosa were detected. The protein express of Bcl-2 of intestinal mucosa was analyzed by immunochemistory.
RESULTIntestinal /R resulted in histopathological changes in intestinal mucosa, increased Chiu's scores, apoptosis index, IWC and MDA content, and reduced SOD activity and the protein expression of Bcl-2 significantly (P < 0.01). The pretreatment of SND could attenuate the above changes significantly (P < 0.01), but there was no significant difference for the above variables between SND low dose group and SND high dose group (P > 0.05). Apoptosis index was significantly negatively correlative to SOD activity in model group and two SND groups. There were significantly negative correlation between apoptosis index and protein expression of Bcl-2 in model group and SND low dose group.
CONCLUSIONSND can attenuate intestinal mucosa injury following intestinal U/R, which is related to reducing intestinal mucosa cell apoptosis by removing oxygen free radical and upregulating the protein expression of Bcl-2.
Aconitum ; chemistry ; Animals ; Apoptosis ; drug effects ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Female ; Ginger ; chemistry ; Glycyrrhiza uralensis ; chemistry ; Intestinal Mucosa ; metabolism ; pathology ; Male ; Plants, Medicinal ; chemistry ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; metabolism ; pathology
6.Role of NF-kappaB in the induction of delayed preconditioning in rat heart by pretreatment with Sini decoction.
Ying LIU ; Wei-Kang WU ; Chen CHEN ; Ke-Xuan LIU
China Journal of Chinese Materia Medica 2005;30(6):453-456
OBJECTIVEThe present study was designed to investigate whether Sini decoction (SND) containing several chinese medicinal herbs, could induce delayed preconditioning-like effect in rat heart and the possible mechanism.
METHODIn anesthetized open-chest Sprague Dawley rats, left anterior descending coronary artery (LAD) was occluded for 1 h and reperfused vided into four groups: Sham ( n = 8) group in which the surgical procedure was identical to other groups,but the LAD ligature was not ligated; I/R group (n = 8) in which the LAD of rat hearts was subjected to the index occlusion; IPC/SWOP group (n = 8) in which the LAD was occluded for three cycles of 5-minute episodes of preconditioning ischemia followed by 5-minute episodes of reperfusion 24 hours prior to the index occlusion; SND group (n = 8) in which rats were pretreated with Sini decoction (5 g x kg(-1) d(-1)) for three days, the last treatment was pretreated 24 h before the index occlusion. Myocardial infarct size, the activity of superoxide dismutase( SOD) and the content of malondialdehyde (MDA) in myocardial tissue were measured. The mRNA expression of MnSOD and Cu-ZnSOD were detected by RT-PCR. The expression of P65 subunit of NF-kappaB was determined by immunohistochemistry assay.
RESULTAs compared with I/R group, myocardial infarct size and the amount of MDA in myocardial tissue were significantly decreased, the activity of SOD and the mRNA expression of MnSOD were increased, the translocation of NF-kappaB was detected and furthermore the expression of NF-kappaB was significantly elevated in SND group as well as in IPC/SWOP group. The mRNA expression of Cu-ZnSOD was lowered in I/R, SND and IPC/SWOP groups, and there was no significant difference among these groups.
CONCLUSIONThe results suggest that Sini decoction can induce delayed preconditioning-like effect in rat heart and the activation of NF-kappaB may play an important role in it.
Animals ; Drugs, Chinese Herbal ; pharmacology ; Ischemic Preconditioning, Myocardial ; methods ; Male ; Myocardial Reperfusion Injury ; metabolism ; pathology ; NF-kappa B ; metabolism ; physiology ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; biosynthesis ; genetics
7.Effects of extracellular ATP on GFAP expression after spinal cord injury in rats
Yan-Chao MA ; Jun QIAN ; Zheng-Yi SUN ; Xue-Wen KANG ; Hai-Hong ZHANG ; Shuan-Ke WANG
Chinese Journal of Neuromedicine 2008;7(10):988-991
Objective To investigate the effects ofextracellular adenosine triphosphate (ATP)on the expression of glial fibrillary acidic protein (GFAP) and the recovery of motor function after spinalcord injury in rats. Methods Six rats were selected as the normal control group randomly from 66adult healthy Wistar rats, and the rest 60 rats were divided into 2 groups after contusion injury wasperformed by the weight drop method with Allen impactor: group A (ATP group) and group B (controlgroup), and every group contained 30 rats. At day 1, 3, 7, 14, and 28 after injury, the rats were killed, andthe expression of GFAP was detected with immunohistochemistry. The expression of GFAP in theadjacent area was semi-quantitatively analyzed with a computer image analysis system; the recovery ofmotor function after spinal cord injury was assessed with improved Tarlov scores. Results Theexpression of GFAP was significantly higher in group A than in Group B after spinal cord injury in rats(P<0.05), and significant difference was also revealed in the expression of GFAP between the 2 groups atday 7, 14 and 28 after injury (P<0.05). At day 14 and 28, Tarlov scores were significantly higher in groupA than in group B (P<0.05). Conclusion Extracellular ATP can promote the expression of GFAP inspinal cord, and may be helpful to the recovery of motor function after spinal cord injury in rats.
8.Efficacy and safety of telbivudine alone and combined with adefovir for the treatment of nucleos(t)ide-naive chronic hepatitis B in patients with high-level hepatitis B virus load.
Na YAO ; Chunfu WANG ; Zhuoran YU ; Ke ZHAO ; Wen KANG ; Qing LIU ; Bianli DANG ; Yongtao SUN
Chinese Journal of Hepatology 2015;23(4):250-253
OBJECTIVETo compare the efficacy of telbivudine monotherapy and telbivudine combination therapy with adefovir in patients with nucleos(t)ide-naive chronic hepatitis B, high-level hepatitis B virus (HBV) load and hepatitis B e antigen (HBeAg)-positivity, and to explore the relationship between treatment regimen adherence and treatment outcomes.
METODSA retrospective study was performed with 123 HBeAg-positive, high-level viral load (HBV DNA≥10(7) copies/ml), nucleos(t)ide-naive chronic hepatitis B patients. Fifty-three of the patients received combination therapy with telbivudine and adefovir dipivoxil,while 70 patients received the telbivudine monotherapy. All patients were tested for rates of conversion to HBV DNA-negative status,alanine aminotransferase (ALT) normalization, HBeAg seroconversion, drug resistance, and side effects at treatment weeks 12, 24, and 48. Treatment regimen adherence was assessed through self-reporting, and interviews were used to explore the relationships to treatment outcomes. The chisquare test, t test and Fisher's exact test were used for statistical analyses.
RESULTSThe rates of HBV DNA negative conversion in the combination group at treatment weeks 12, 24 and 48 were 62.3% (33/53), 88.7% (47/53) and 94.3% (50/53) and were significantly different from those in the monotherapy group at weeks 12 and 24.The rates of ALT normalization were significantly different between the two groups at treatment week 12 (94.3% vs. 77.1%). The rate of HBeAg seroconversion in the combination group at treatment week 48 was 39.6%, and significantly different than that of the monotherapy group. The rates of drug-resistance in the combination and monotherapy groups at treatment week 48 were 3.8% and 11.4%, and the proportion of non-adherence to the treatment regimen was 53.3%, which significantly affected treatment outcome. No side effects occurred in either treatment group.
CONCLUSIONTelbivudine combination treatment with adefovir was more effective than telbivudine monotherapy and elicited a low drug resistance rate in nucleos(t)idenaive chronic hepatitis B patients with high-level HBV load and HBeAg-positivity. Adherence to the therapy regimen was a key factor influencing treatment outcomes.
Adenine ; analogs & derivatives ; Alanine Transaminase ; Drug Therapy, Combination ; Hepatitis B e Antigens ; Hepatitis B virus ; Hepatitis B, Chronic ; Humans ; Organophosphonates ; Retrospective Studies ; Thymidine ; analogs & derivatives ; Treatment Outcome ; Viral Load
9.Expression of estrogen receptor alpha in the testis of infertile men with spermatogenic arrest.
Gang WANG ; Shou-yi GU ; Kang-ning CHEN ; Zhen-xian WANG ; Ting-jiang LIU ; Ke-jian SUN ; Ying-wei ZHAO ; Fu-zhen SUN ; Xiang-yun YIN
National Journal of Andrology 2011;17(1):27-31
OBJECTIVETo explore the association of spermatogenic arrest with the expression of estrogen receptor alpha (ERalpha) in human testes.
METHODSWe examined the testicular biopsy specimens of 120 infertile men by HE staining, detected the expression of ERalpha in the specimens of those with spermatogenic arrest by the two-step immunohistochemical method, and compared the results with those of 10 healthy men.
RESULTSOf the 120 specimens from the infertile men, 31 (25.8%) met the diagnostic criteria of spermatogenic arrest. In the testis tissue of normal men, ERalpha expressed in Sertoli, myoid and Leydig cells, but not in spermatogenic cells, while in the testis tissues of those with spermatogenic arrest, ERalpha expressed lowly in Sertoli, myoid and Leydig cells, with statistically significant differences in immunostaining intensity between the two groups (P < 0.05).
CONCLUSIONAndrogen receptor (AR) and ERalpha may play a coordinating role in facilitating spermatogenesis. Spermatogenic arrest may be related to a complex series of disorders in cell signal transduction involving AR, ERalpha and HSP90.
Adult ; Case-Control Studies ; Estrogen Receptor alpha ; metabolism ; Humans ; Infertility, Male ; etiology ; metabolism ; pathology ; Male ; Spermatogenesis ; Testis ; metabolism ; pathology ; Young Adult
10.Peroxisome proliferator-activated receptor alpha agonist attenuates oxidized-low density lipoprotein induced immune maturation of human monocyte-derived dendritic cells.
Hong-Yu SHI ; Jun-Bo GE ; Wei-Yi FANG ; Kang YAO ; Ai-Jun SUN ; Rong-Chong HUANG ; Qing-Zhe JIA ; Ke-Qiang WANG ; Yun-Zeng ZOU ; Xue-Tao CAO
Chinese Medical Journal 2008;121(17):1747-1750
Atherosclerosis
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etiology
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prevention & control
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Cytokines
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secretion
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Dendritic Cells
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drug effects
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immunology
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Endocytosis
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Fenofibrate
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pharmacology
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Humans
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Immunophenotyping
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Lipoproteins, LDL
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toxicity
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Monocytes
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cytology
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PPAR alpha
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agonists
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physiology