2.Improvement of transcriptional activity of hTERT promoter by SV40 enhancer.
Wei-ming ZHANG ; Li-ying XUE ; Yao XU ; Jun XING ; Xin GENG ; Dong WANG ; Yan-yun LI
Chinese Journal of Pathology 2006;35(11):691-693
Base Sequence
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Cell Line, Tumor
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Enhancer Elements, Genetic
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genetics
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HT29 Cells
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Humans
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Luciferases
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genetics
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metabolism
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Molecular Sequence Data
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Plasmids
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genetics
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Polymerase Chain Reaction
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Promoter Regions, Genetic
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genetics
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Recombinant Fusion Proteins
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genetics
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metabolism
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Simian virus 40
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genetics
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Telomerase
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genetics
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Transcription, Genetic
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Transfection
3.Differential proteomic expression in human liver cells stimulated by hydroquinone.
Li JU ; Shu-Zhi ZHANG ; Ran ZHAO ; Geng-Dong YAO
Chinese Journal of Industrial Hygiene and Occupational Diseases 2006;24(11):658-661
OBJECTIVETo explore the differential proteomic expression in human liver cells L-02 after exposure to HQ.
METHODSSubcultured L-02 cells were treated by HQ for 24 h at a 1 x 10(-4) mol/L concentration and a blank group was set as the control. Immediately after the treatment, total cellular proteins were extracted and separated by 2-DE, and the images were analyzed by PDQuest software. The experiment was totally repeated 3 times with 3 repetitions for each group every time. The well repeated spots were identified by MALDI-TOF MS and then searched in NCBI human protein database with Mascot.
RESULTSAbout 1,000 spots per gel were found. Compared with the control group, 17, 18 and 24 spots were significantly altered in 3 separate experiments. The 4 well repeated spots were identified by MALDI-TOF MS as Rho GDP dissociation inhibitor GDI alpha, 6-phosphogluconolactonase, erbB3 binding protein EBP1 and lamin A/C, isoform 1 precursor. They were involved in cell skeleton, signal transduction and energy metabolization in functional classification.
CONCLUSIONHydroquinone can change the protein expression in liver cells, which provides clues for exploring the toxic mechanism.
Cell Line ; Electrophoresis, Gel, Two-Dimensional ; Hepatocytes ; drug effects ; metabolism ; Humans ; Hydroquinones ; toxicity ; Mass Spectrometry ; Proteomics ; Reproducibility of Results
4.Impact of 1.8GHz Radiofrequency Electromagnetic Fields on Gene Expression Profiles in Rat Neurons
Shu-Zhi ZHANG ; Geng-Dong YAO ; De-Qiang LU ; Huai CHIANG ; Zheng-Ping XU
Journal of Preventive Medicine 2009;21(7):3-6
Objective To investigate the changes of gene expression in rat neurons induced by 1.8 GHz radiofrequency electromagnetic fields (RF EMF) and to screen RF EMF-responsive genes. Methods The rat primary cultured neuronal cells were divided into two groups, the radiation group and control group, from which the total RNA was extracted immediately and purified after intermittently (5min on/10min off) exposed or U34 array was applied to detect the changes of gene expression in rat neurons. Results Among 1200 candidate genes, 24 up-regulated and 10 down-regulated genes which are associated with multiple cellular functions (cytoskeleton, signal transduction pathway, metabolism, etc.) after functional classification were found by using Affymetrix microarray suite software 5.0. Although the changes in gene expression were less than 2 folds, they had statistical significance (P<0.01). Conclusion RF radiation of 1.8GHz induce the changes of many genes transcription in rat neurons, some of which indicate the negative effects of RF radiation on neurons.
5.Effect of 1.8 GHz radiofrequency electromagnetic fields on gene expression of rat neurons.
Shu-zhi ZHANG ; Geng-dong YAO ; De-qiang LU ; Huai CHIANG ; Zheng-ping XU
Chinese Journal of Industrial Hygiene and Occupational Diseases 2008;26(8):449-452
OBJECTIVETo investigate the changes of gene expression in rat neuron induced by 1.8 GHz radiofrequency electromagnetic fields (RF EMF) to screen for RF EMF-responsive genes and the effect of different exposure times and modes on the gene expression in neuron.
METHODSTotal RNA was extracted immediately and purified from the primary culture of neurons after intermittent exposed or sham-exposed to a frequency of 1.8 GHz RF EMF for 24 hours at an average special absorption rate (SAR) of 2 W/kg. Affymetrix Rat Neurobiology U34 array was applied to investigate the changes of gene expression in rat neuron. Differentially expressed genes (Egr-1, Mbp and Plp) were further confirmed by semi-quantitative revere transcription polymerase chain reaction (RT PCR). The expression levels of Egr-1, Mbp and Plp were observed at different exposure times (6, 24 h) and modes (intermittent and continuous exposure).
RESULTSAmong 1200 candidate genes, 24 up-regulated and 10 down-regulated genes were found by using Affymetrix microarray suite software 5.0 which are associated with multiple cellular functions (cytoskeleton, signal transduction pathway, metabolism, etc.) after functional classification. Under 24 h and 6 h intermittent exposure, Egr-1 and Plp in experiment groups showed statistic significance (P < 0.05) compared with the control groups, while expression of Mbp did not change significantly (P > 0.05). After 24 h continuous exposure, Egr-1 and Mbp in experiment groups showed statistic significance (P < 0.05) compared with the control group, while expression of Plp did not change significantly (P > 0.05). Under the same exposure mode 6 h, expression of all the 3 genes did not change significantly. Different times (6, 24 h) and modes (intermittent and continuous exposure) of exposure exerted remarkable different influences on the expression of Egr-1, Mbp, Plp genes (P < 0.01).
CONCLUSIONThe changes of many genes transcription were involved in the effect of 1.8 GHz RF EMF on rat neurons; Down-regulation of Egr-1 and up-regulation of Mbp, Plp indicated the negative effects of RF EMF on neurons; The effect of RF intermittent exposure on gene expression was more obvious than that of continuous exposure; The effect of 24 h RF exposure (both intermittent and continuous) on gene expression was more obvious than that of 6 h (both intermittent and continuous).
Animals ; Cells, Cultured ; Dose-Response Relationship, Radiation ; Down-Regulation ; radiation effects ; Electromagnetic Fields ; Neurons ; metabolism ; radiation effects ; Rats ; Up-Regulation ; radiation effects
6.Effects of acrylamide on DNA damage in human keratinocytes.
Xiao-xia MA ; Geng-dong YAO ; Hao CHENG ; Qun-li ZENG ; Qing CHEN
Chinese Journal of Industrial Hygiene and Occupational Diseases 2003;21(2):96-98
OBJECTIVETo investigate the toxic and DNA damaging effect of acrylamide (AA) on human keratinocytes and its mechanism.
METHODS(1) After the keratinocyte cell line HaCaT cells were exposed to AA with different concentrations for 44 hours, cell survival rate was detected by MTT method. (2) The effects of DNA damage of exposed cells were detected by comet assay. (3) After treating the cells with 2.00 mmol/L of AA plus 0.50 mmol/L of 1-aminobenzotriazole (1-ABT), an inhibitor of cytochrome P-450 enzymes (CYP-450), for 4 hours, the relationship between DNA damage and CYP-450 was studied.
RESULTS(1) Cytotoxicity measurement of AA showed that cell survival rate decreased significantly after 44-hour treatment. (2) Cytotoxicity was not detected after 4-hour AA treatment, but significant DNA damage was observed in all treatment groups, and the degree of damage increased with the concentration of AA. Moreover, the tail lengths of comet cells were in dose-effect relationship. As for cells treated by 1-ABT with 2 mmol/L AA, comet rate and tail length were 15.4% and (8.2 +/- 2.0) micro m respectively, which were decreased significantly (P < 0.01) when compared with 2 mmol/L AA treatment group [80.6% and (44.3 +/- 4.0) micro m].
CONCLUSIONSAcrylamide has significant cytotoxicity and genotoxicity on HaCaT cells. AA-induced DNA damage may be related to the oxidative metabolite(s) of AA through CYP-450.
Acrylamide ; toxicity ; Cells, Cultured ; Cytochrome P-450 Enzyme Inhibitors ; DNA Damage ; Dose-Response Relationship, Drug ; Humans ; Keratinocytes ; drug effects ; enzymology
7.Effect of 1.8 GHz radiofrequency electromagnetic fields on the expression of microtubule associated protein 2 in rat neurons.
Ran ZHAO ; Shu-zhi ZHANG ; Geng-dong YAO ; De-qiang LU ; Huai JIANG ; Zheng-ping XU
Chinese Journal of Industrial Hygiene and Occupational Diseases 2006;24(4):222-225
OBJECTIVETo investigate the changes of gene expression in rat neurons induced by 1.8 GHz radiofrequency electromagnetic fields (RF EMF) and to screen for the RF EMF-responsive genes.
METHODSNewly-born SD rats in 24 hours were sacrificed to obtain cortex and hippocampus neurons. The cells were divided randomly into two groups: the experiment group (the irradiation group) and the control group (the false irradiation group). In the irradiation group, after twelve days' culture, neurons were exposed to 1.8 GHz RF EMF modulated by 217 Hz at a specific absorption rate (SAR) of 2 W/kg for 24 hours (5 minutes on/10 minutes off) while in the false control group, the neurons were put in the same waveguide as in the irradiation group, but were not exposed to any irradiation. The total RNA was isolated and purified immediately after exposure. The affymetrix rat neurobiology U34 assay was used for detecting the changes in gene expression profile according to the manufacturer's instruction. RF EMF-responsive candidate gene was confirmed by using ribonuclease protection assay (RPA).
RESULTSAmong 1200 candidate genes, the expression levels of 34 genes were up or down regulated. Microtubule associated protein 2 (Map2) gene was selected as the candidate and subjected to further analysis. RPA data clearly revealed that Map2 was statistically significantly up-regulated after neurons were exposed to the RF EMF (P < 0.05).
CONCLUSIONThe modulation of gene expression and function of Map2 as a neuron specific cytoskeleton protein is crucial to maintain the normal framework and function of neurons. The finding that 1.8 GHz RF EMF exposure increases the expression of Map2 might indicate some unknown effects of RF EMF on neurons.
Animals ; Animals, Newborn ; Cell Phone ; Cells, Cultured ; Dose-Response Relationship, Radiation ; Down-Regulation ; Electromagnetic Fields ; Female ; Gene Expression ; radiation effects ; Male ; Microtubule-Associated Proteins ; biosynthesis ; genetics ; Neurons ; metabolism ; radiation effects ; Radio Waves ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Up-Regulation
8.Curative effect observation of erlotinib plus temozolomide for recurrence/progression in patients with EGFR gene mutation in NSCLC with brain metastases after whole brain radiotherapy
Qiao YANG ; Yao LIU ; Hui QIU ; Yi GENG ; Xiaohong YAN ; Jie HOU ; Cuihua CUI ; Longke DONG
Journal of International Oncology 2019;46(5):257-261
Objective To observe the clinical efficacy and safety of erlotinib plus temozolomide for recurrence/progression patients with epidermal growth factor receptor (EGFR) gene mutation in non-small cell lung cancer (NSCLC) with brain metastases after whole brain radiotherapy.Methods A total of 68 EGFR gene mutation NSCLC patients with brain metastases of intracranial recurrence/progression after whole brain radiotherapy were selected from August 2013 to June 2018 in Baoji Central Hospital of Shaanxi Province and Xintai People's Hospital of Shandong Province.All the patients were randomly divided into erlotinib group and combined treatment group (erlotinib combined with temozolomide) using random number table method.The patients in erlotinib group (34 cases) were treated with oral erlotinib 150 mg/d until progression or unacceptable adverse reaction,and the patients in combined treatment group (34 cases) were given erlotinib and oral temozolomide 150 mg/(m2 · d) for 1-5 day,every 28 days was a cycle,temozolamide for 6 cycles.Comparison was made on curative effects and occurrence condition of adverse reactions between the two groups.Results The overall response rates in the erlotinib group and combined treatment group were 11.8% (4/34)and 32.4% (11/34) respectively,and the disease control rates in the two groups were 35.3% (12/34) and 64.7% (22/34) respectively,with significant differences (x2 =4.191,P =0.041;x2 =5.882,P =0.015).The median progression-free survival in the erlotinib group and combined treatment group were 3.22 months and 5.29 months respectively,and the median overall survival in the two groups were 5.60 months and 7.90 months respectively,with significant differences (x2 =9.269,P =0.002;x2 =11.005,P =0.001).The incidence of nausea and vomiting in combined treatment group was significantly higher than that in erlotinib group [67.6% (23/34) vs.14.7% (5/34)],with a significant difference (x2 =19.671,P < 0.001),but there were no significant differences in the incidences of other adverse reactions (all P > 0.05).The patients in the two groups had no more than grade Ⅲ of adverse reactions.Conclusion The curative effect of erlotinib combined with temozolomide is better in the treatment of recurrence/progression patients with EGFR gene mutation in NSCLC with brain metastases after whole brain radiotherapy,with mild adverse reactions and good patients' tolerance.
9.Molecular and virulence characteristics of Staphylococcus aureus isolated from neonates
Wenjing GENG ; Shixiao DONG ; Fei JIN ; Wei SHI ; Kaihu YAO ; Mingyan HEI
Chinese Journal of Microbiology and Immunology 2020;40(6):429-436
Objective:To analyze the molecular and virulence characteristics of Staphylococcus aureus ( S. aureus) isolated from neonates. Methods:A total of 189 S. aureus isolates were collected from Beijing Children′s Hospital from January 2013 to October 2019 and analyzed by multilocus sequence typing (MLST), staphylococcal cassette chromosome mec (SCC mec) typing and Spa typing. Panton-valentine leucocidin (PVL)-encoding gene ( pvl) and 21 superantigen virulence genes were also detected. Results:The 189 S. aureus strains were isolated from respiratory secretions ( n=125), pus ( n=55), blood ( n=8) and pleural effusion ( n=1). There were 98 methicillin-susceptible S. aureus (MSSA) belonging to 42 MSSA clones and 91 methicillin-resistant S. aureus (MRSA) belonging to 26 MRSA clones. ST188-t189 and ST59-SCC mecⅣa-t437 were the predominant MSSA and MRSA clones accounting for 11% and 53%, respectively. The prevalence of pvl gene in MRSA isolates was significantly higher than that in MSSA isolates (32% vs 10%, P<0.01). There were 166 isolates (88%) carrying at least one superantigen virulence gene and among the 21 genes, seq and seb were the most common genes accounting for 47% and 43%, respectively. The most common superantigen genotype was seb- sek- seq. The positive rates of superantigen genes in MRSA and MSSA isolates were 85% (77/91) and 90% (88/98), respectively ( P>0.05). Conclusions:The main clones of MRSA and MSSA were different in neonates. ST59-SCC mecⅣa-t437 was the most predominant MRSA clone, while ST188-t189 was the most predominant MSSA clone. MSSA clones were more dispersed. The prevalence of pvl gene in MRSA was higher than that in MSSA. No significant difference in the prevalence of superantigen genes was observed between MRSA and MSSA.
10.Awake craniotomy and intraoperative language cortical mapping for eloquent cerebral glioma resection: preliminary clinical practice in 3.0 T intraoperative magnetic resonance imaging integrated surgical suite.
Jun-feng LU ; Jie ZHANG ; Jin-song WU ; Cheng-jun YAO ; Dong-xiao ZHUANG ; Tian-ming QIU ; Xiu GONG ; Geng XU ; Ying MAO ; Liang-fu ZHOU
Chinese Journal of Surgery 2011;49(8):693-698
OBJECTIVESTo evaluate preliminary clinical experience for combining awake craniotomy and intraoperative language brain mapping within the integrated 3.0 T intraoperative magnetic resonance imaging (iMRI) suite.
METHODSFrom December 2010 to April 2011, 11 right hand-dominant patients with left glioma were involved in, or adjacent to, eloquent cortex was carried out awake craniotomies with cortical stimulation within an integrated 3.0 T iMRI suite. Aphasia battery of Chinese was used to test the language function before the operation. During the procedure, after the occipital, temporal, and supraorbital nerves were blocked by the anesthesiologists, the head was fixed with a custom high-field MRI-compatible head holder. The skull and dura was opened as usual and language brain mapping was then performed. Language testing followed a set protocol: counting numbers from 1 to 50, naming objects, reading single words. Resection of the tumor was guided by neuronavigation system and continued until eloquent areas were encountered or the margin of assessment was reached. An interdissection MRI was acquired to evaluate the glioma removal in a movable MRI scanner after minimal draping. Meanwhile, adverse effects caused by electrical stimulation and iMRI were recorded. The follow-up speech tests were assessed on 7th day and 1 month at least after the operation.
RESULTSThe combined use of 3.0 T iMRI and awake craniotomy was performed safely in all patients. No adverse effects were reported. The duration of surgery was prolonged by 2 to 4 h. The patients' perception of iMRI during surgery was favorable. First-look MRI studies led to further resection attempts in 6/11 cases as well as a 3/11 increase in the number of gross-total resections. One week after surgery, baseline language function worsened in 4 cases. However, no patients had a persistent language deficit one month after surgery.
CONCLUSIONSAwake craniotomy and direct cortical electrical stimulation can be performed safely and effectively within a 3.0 T iMRI suite. The combination of high-field iMRI and awake craniotomy may facilitate safe removal of eloquent glioma.
Adult ; Aged ; Anesthesia ; methods ; Brain Neoplasms ; surgery ; Cerebral Cortex ; surgery ; Craniotomy ; methods ; Female ; Glioma ; surgery ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Monitoring, Intraoperative ; Neuronavigation ; methods ; Wakefulness