1.The inhibitory effect of silencing RAGE gene expression by shRNA on the proliferation of prostate cancer cells
Xinping SUN ; Chunyan MA ; Yulian JIAO ; Yunyun QU ; Min ZHU ; Xiaowen LIU ; Jie XU ; Yueran ZHAO
Tumor 2010;(3):199-204
Objective:To construct a specific small hairpin RNA (shRNA) expressing vectors against human receptor for advanced glycation end product (RAGE) gene and study its inhibitory effect on the proliferation of androgen-independent prostate cancer cells DU145. Methods:Four RAGE specific oligonucleotides were designed and synthesized. These oligonucleotides were annealed to forill double strand DNA fragments and this fragment was cloned into psi-U6 plasmid. The recombinants were transfected into RAGE-overexpressing sub DU145-2C1 cells. Cellular morphology and transfection efficiency were observed under fluorescence microscope. The inhibitory effect of RAGE shRNA construct on RAGE mRNA and protein expression was examined with semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blotting, respectively. The cellular proliferation was detected with cell counting kit-8 (CCK-8). Scratch test was used to observe the migration of DU145 cells.Results:RAGE shRNA expression plasmids were successfully constructed and transfected into sub DU145-2C1 cells. It can effectively inhibit the expression of RAGE mRNA (P<0.05). The inhibitory effects of shRNA RAGE-1 (R1) was the most stronger. The RAGE mRNA expression was inhibited by 84% and RAGE protein expression was inhibited by 27%. Compared with negative control, the proliferation potential was significantly decreased in shRNA RAGE-transfected cells. The cell migration capability had no significant changes. Conclusion:RAGE shRNA effectively inhibited the expression of RAGE mRNA and protein and suppressed the proliferation of DU145 cells in vitro.
2.Study on changes of vascular endothelium secreted factors after oral fatty meal test in elderly diabetic patients
Minglong LI ; Ping YANG ; Bo LIANG ; Yulian JIAO ; Qiu LI ; Haiyan CHEN ; Yinyin LI
Chinese Journal of Geriatrics 2008;27(12):892-896
Objective To investigate the dynamic changes of vascular endothelium secreted factors after oral fatty meal test and the correlation of the factors with blood lipid in elderly diabetic patients. Methods Thirty-six elderly diabetic patients (diabetic group) and twenty heahhy elderly subjects(control group) were selected into the study and received oral fatty meal test for 6 hours. Diabetic group was divided into three subgroups, including fasting hypertriglyceridemia subgroup, postprandial hypertriglyceridemia subgroup and postprandial normotriglyceridemia subgroup. Serum nitric oxide (NO), endothelin-1 (ET-1), plasminogen activator inhibitor-1 (PAl-1) and tissue plasminogen activator (t-PA) were measured before and after oral fatty meal test. Results (1) The level of serum NO was significantly increased and ET-1 was significantly reduced in control group 2 hours after oral fatty meal test and were returned to basal state 6 hours after the test. But in diabetic group, postprandial serum NO level were decreased and ET-1 were increased gradually and reached to the peak 6 hours after oral fatty meal test. The ratio of NO/ET-1 was lower in diabetic group than that in control group (P< 0.01). There were important differences among fasting hypertriglyceridemia subgroup, postprandial hypertriglyceridemia subgroup and postprandial normotriglyeeridemia subgroup(P<0.05 or<0.01). (2) The level of PAl-1 was increased and t-PA was decreased slightly 4 hours after oral fatty meal test in control and diabetic groups. Compared with control group, PAI-1/ t-PA obviously increased in diabetic group. Meanwhile, PAI-1/t-PA in fasting hypertriglyeeridemia and postprandial hypertriglyceridemia subgroups were significantly higher than that in postprandial normotriglyceridemia subgroup(P<0. 05 or<0.01). (3) In the diabetic group, TG was negatively correlated with NO and t-PA(r=-0.360 P<0.05; r=-0.649, P<0.01) and positively correlated with ET-1 and PAI-1(r=0.421,P<0.01;r=0.520,P<0.01). Conclusions The elderly diabetic patients suffer from the imbalance of vascular endothelium secreted factors. The postprandial abnormal TG metabolism may aggravate the change and further damage the vascular endothelial function.
3.Association of HLA-Cw alleles with systemic lupus erythematosus
Hongsheng SUN ; Yanfeng HOU ; Yuanchao ZHANG ; Yulian JIAO ; Qingrui YANG ; Guangzhi SUN ; Yueran ZHAO
Chinese Journal of Rheumatology 2008;12(10):667-669
Objective To explore the association between HLA-Cw alleles with systemic lupuserythematosus. Methods Polymerase chain reaction-sequence specific primer method was used to analyze thedistribution of HLA-Cw01-08 alleles among 108 patients with SLE and 102 healthy controls. The allelefrequencies was compared between various patient groups and the control population. Results The frequencyof HLA-Cw07 alleles in patients with SLE was significantly increased in patients with SLE. Conclusion Theresults indicate that HLA-Cw07 may be the susceptible alleles or may be closely linked to the susceptiblegenes for SLE.
4.Gene polymorphism of killer cell immunoglobulin-like receptors and HLA-Cw in patients with ankylosing spondylitis
Bingchang ZHANG ; Zhiming LU ; Yun LIU ; Yulian JIAO ; Yueran ZHAO ; Jianfeng LI
Chinese Journal of Laboratory Medicine 2008;31(10):1114-1118
Objective To investigate the alterations in killer cell immunoglobulin-like receptors (KIRs)2D and their specific HLA-Cw ligands in patients with ankylosing spondylitis(AS)and determine whether the changes were correlate to the pathogenesis of AS.Methods Polymerase chain reaction of sequence specific primerB(PCR-SSP)was employed for genotyping the presence or absence of five KIR2D genes(KIR2DL1,2DS1,KIR2DL2,2DL3,2DS2)as well as HLA-Cw01-08 alleles from genomic DNA in 105 individuals with AS,together with 51 individuals with osteoarthritis(OA)and 120 healthy controls.Then HLA-C10-08 was divided into two groups.HLA-Cwasn and HLA-Cwlys to calculate the frequency of KIRID genotype.HLA-Gu alleles and KIR/HLA-Cw genotypes.Results The frequencies of HLA-Cwlys genes were significandy higher in patients with AS(0.269 7)compared with those in OA controls(0.148 2)and healthy controls(0.138 8,P=0.024,P=0.001,respectively).The frequency of KIR2DS1/HLA-Cwlys combination Was also markedly higher in AS group(26.67%)than that in OA controls(11.76%)and healthy controls(13.33%,P=0.039,P=0.018,respectively).Condusion The data suggest that the HLA-Cwlys allele may be associated with genetic susceptibility to AS and moreover.in the existence of HLA-Cwlys.the individuals with KIR2DS1 gene are likely to be at increased risk of AS.
5.Killer cell immunoglobulin-like receptor gene polymorphism in Hashimoto′s thyroiditis patients
Haiqing ZHANG ; Jiajun ZHAO ; Yueran ZHAO ; Ling GAO ; Lin LIAO ; Qingbo GUAN ; Yulian JIAO
Chinese Journal of Microbiology and Immunology 2008;28(5):454-457
Objective To investigate the relationship of the killer cell immunoglobulin-like receptor (KIR) gene polymorphism with Hashimoto′s thyroiditis(HT). Methods One hundred HT patients and 260 randomly matched healthy controls were enrolled to detect the KIR genotype. The genomic DNA were extracted, and 15 selected KIR genes, KIR2DL1-5, KIR3DL1-3, KIR2DS1-5, KIR3DS1 and pseudogene KIR2DP1, were determined by a polymerase chain reaction using sequence-specific primers (PCR-SSP). Results The frequency of KIR2DL5 gene was significantly lower of the patient group than that of the control group (0.200 vs 0.312, RR=0.64, P<0.01). Conclusion There may be an association between pathogenesis of HT and KIR2DL5 gene.
6.A FKBP5 mutation is associated with Paget's disease of bone and enhances osteoclastogenesis.
Bingru LU ; Yulian JIAO ; Yinchang WANG ; Jing DONG ; Muyun WEI ; Bin CUI ; Yafang SUN ; Laicheng WANG ; Bingchang ZHANG ; Zijiang CHEN ; Yueran ZHAO
Experimental & Molecular Medicine 2017;49(5):e336-
Paget's disease of bone (PDB) is a common metabolic bone disease that is characterized by aberrant focal bone remodeling, which is caused by excessive osteoclastic bone resorption followed by disorganized osteoblastic bone formation. Genetic factors are a critical determinant of PDB pathogenesis, and several susceptibility genes and loci have been reported, including SQSTM1, TNFSF11A, TNFRSF11B, VCP, OPTN, CSF1 and DCSTAMP. Herein, we report a case of Chinese familial PDB without mutations in known genes and identify a novel c.163G>C (p.Val55Leu) mutation in FKBP5 (encodes FK506-binding protein 51, FKBP51) associated with PDB using whole-exome sequencing. Mutant FKBP51 enhanced the Akt phosphorylation and kinase activity in cells. A study of osteoclast function using FKBP51V55L KI transgenic mice proved that osteoclast precursors from FKBP51V55L mice were hyperresponsive to RANKL, and osteoclasts derived from FKBP51V55L mice displayed more intensive bone resorbing activity than did FKBP51WT controls. The osteoclast-specific molecules tartrate-resistant acid phosphatase, osteoclast-associated receptor and transcription factor NFATC1 were increased in bone marrow-derived monocyte/macrophage cells (BMMs) from FKBP51V55L mice during osteoclast differentiation. However, c-fos expression showed no significant difference in the wild-type and mutant groups. Akt phosphorylation in FKBP51V55L BMMs was elevated in response to RANKL. In contrast, IκB degradation, ERK phosphorylation and LC3II expression showed no difference in wild-type and mutant BMMs. Micro-CT analysis revealed an intensive trabecular bone resorption pattern in FKBP51V55L mice, and suspicious osteolytic bone lesions were noted in three-dimensional reconstruction of distal femurs from mutant mice. These results demonstrate that the mutant FKBP51V55L promotes osteoclastogenesis and function, which could subsequently participate in PDB development.
Acid Phosphatase
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Animals
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Asian Continental Ancestry Group
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Bone Diseases, Metabolic
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Bone Remodeling
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Bone Resorption
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Femur
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Humans
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Mice
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Mice, Transgenic
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Osteitis Deformans*
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Osteoblasts
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Osteoclasts
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Osteogenesis
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Phosphorylation
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Phosphotransferases
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Tacrolimus Binding Proteins
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Transcription Factors
7.Chronic ethanol feeding impairs AMPK and MEF2 expression and is associated with GLUT4 decrease in rat myocardium.
LiYong CHEN ; FuRong WANG ; XiangLan SUN ; Jing ZHOU ; Ling GAO ; YuLian JIAO ; XiaoLei HOU ; ChengYong QIN ; JiaJun ZHAO
Experimental & Molecular Medicine 2010;42(3):205-215
Chronic and heavy alcohol consumption is one of the causes of heart diseases. However, the effects of ethanol on insulin sensitivity in myocardium has been unclear. To investigate the effects of ethanol on the expression of AMP-activated protein kinase (AMPK), myocyte enhancer factor 2 (MEF2) and glucose transporter 4 (GLUT4), all of which are involved in the regulation of insulin sensitivity, in the myocardium, we performed three parts of experiments in vivo and in vitro. I: Rats were injected with 5-amino-4-imidazolecarboxamide ribonucleotide (AICAR, 0.8 mg.kg(-1)) for 2 h. II: Rats received different dose (0.5, 2.5 or 5 g.kg(-1).d(-1)) of ethanol for 22-week. III: Primary neonatal rat cardiomyocytes were isolated and treated with or without 100 mM ethanol or 1 mM AICAR for 4 h. The cardiac protein and mRNA expression of AMPKalpha subunits, MEF2 and GLUT4 were observed by western-blotting and RT-PCR, respectively. Serum TNFalpha levels were assessed by ELISA. The results showed chronic ethanol exposure induced insulin resistance. Ethanol decreased the mRNA levels of AMPKalpha1 and alpha2, the protein levels of total- and phospho-AMPKalpha in cardiomyocytes. Similarly, ethanol showed inhibitory effects on both the mRNA and protein levels of MEF2A and 2D, and GLUT4 in a dose-response-like fashion. Correlation analysis implied an association between phospho-AMPKalpha and MEF2A or MEF2D, and between the levels of MEF2 protein and GLUT4 transcription. In addition, ethanol elevated serum TNFalpha level. Taken together, chronic ethanol exposure decreases the expression of AMPKalpha and MEF2, and is associated with GLUT4 decline in rat myocardium.
AMP-Activated Protein Kinases/genetics/*metabolism
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Aminoimidazole Carboxamide/analogs & derivatives/pharmacology
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Animals
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Enzyme Activation/drug effects
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Ethanol/*administration & dosage/*pharmacology
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Feeding Behavior/*drug effects
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Gene Expression Regulation/drug effects
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Glucose Transporter Type 4/genetics/*metabolism
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Insulin/pharmacology
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Insulin Resistance
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Male
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Myocardium/*enzymology
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Myogenic Regulatory Factors/antagonists & inhibitors/genetics/*metabolism
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Protein Isoforms/antagonists & inhibitors/genetics/metabolism
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RNA, Messenger/genetics/metabolism
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Rats
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Rats, Wistar
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Ribonucleotides/pharmacology
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Time Factors
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Tumor Necrosis Factor-alpha/blood
8.Investigation and analysis of the needs of young students for popularizing science about nuclear and radiation
Hongjun ZHAO ; Fang WANG ; Yulian LIU ; Haiyun WANG ; Ling JIAO
Chinese Journal of Radiological Medicine and Protection 2021;41(10):746-751
Objective:To be aware of the needs of the young students for nuclear and radiation science popularization, and to provide scientific basis for accurate science popularization.Methods:A simple random sampling method was used to select 1 primary school, 5 middle schools and 2 universities in the Beijing-Tianjin-Hebei region in December 2020. Questionnaires were distributed through teachers. In addition, convenience sampling method was used to distribute questionnaires in friend circle and other areas to expand the survey scope, with a total of 1 345 respondents. SPSS was used to conduct statistical analysis on the basic information of the respondents, the understanding and concern of nuclear and radiation science popularization and the demand for nuclear and radiation science popularization.Results:A total of 1 120 valid questionnaires were collected, of which 52.4% mainly remained at the conceptual level for the cognition of radiation, 52.2% occasionally paid attention to nuclear and radiation science popularization, 65.3% and 41.3% paid attention to life reference and hobbies, respectively. Radiation protection and its sources and effects received high concern, accounting for 72.6% and 68.3% respectively. Illustration and short video were popular science forms of young students, making up 45.7% and 44.3%, respectively. The students of different genders differed in radiation cognition, degree of concern, purpose of concern and content demand for radiation protection science popularization, and the differences are statistically significant( χ2=10.017, 26.859, 56.237, 17.305, P<0.05). Conclusions:Nuclear and radiation science popularization should consistent with the law of public demand, accurately locate the demand characteristics of young students, and consider the characteristics of different genders, concerns over radiation protection, treatment and damage knowledge from the point of life and fun, so as to improve the public′s attention, enhance the national nuclear science culture, and create a good nuclear safety culture atmosphere.
9.Effect of high dose irradiation on thermoluminescence detector
Shuzhou RUAN ; Zhixin ZHAO ; Wenyi ZHANG ; Kaijun SU ; Xinxin PANG ; Fang WANG ; Yulian LIU ; Menghui HUO ; Changxin YAN ; Ling JIAO
International Journal of Biomedical Engineering 2019;42(4):342-345
Objective To investigate effect of high dose irradiation on the performances of thermolumines-cence detectors (LiF:Mg, Cu, P). Methods The high-dose irradiated thermoluminescence detector was annealed by a thermoluminescence annealing furnace until the annealing was completed, and then the annealed thermoluminescent detector was irradiated 0.5Gy by 137Cs irradiator to verify the accuracy of the thermoluminescentdetector. Results The thermoluminescence detector after high-dose irradiation could not be completely annealed under the temperature condition of 240 ℃, and it could be completely annealed at a high temperature as 400℃. After 0.5 Gy irradiation by 137Cs irradiator, the measurement results of the annealed thermoluminescence detector were significantly smaller, and the dose response and dispersion of the detector were also changed significantly. Conclusions After a more than 5 Gy irradiation, the crystal structure of the thermoluminescence detector has changed, and a high temperature peak above 240 ℃ has appeared, which leads to the failure to completely anneal at normal temperature. Therefore, the thermoluminescence detector can no longer be used for dosimetry after high-dose irradiation.