1.EXPRESSION AND DISTRIBUTION OF AROMATASE mRNA IN THE BRAIN OF THE MOUSE
Acta Anatomica Sinica 1953;0(01):-
Objective\ To exam the expression and the distribution of the aromatase mRNA in the brain of the mouse. Methods\ RNA dot\|blotting as well as in situ hybridization technique were used. Results\ (1)There were aromatase specific mRNA expression in the brain tissue during the period from E16 to P300,the highest levels of mRNA were detected at postnatal 6 days,and the lowest levels were found at adulthood.(2)The location of the aromatase mRNA was confined to neuronal(but not glial)cell bodies and their processes.(3)The mainly distribution of aromatase mRNA was detected in the regions of the cerebral cortex,thalamus,hypothalamus and limbic system.Many heavily labeled cells were found in the layer of pyramidal cells of cerebral cortex,medial preoptic area.medial septal nucleus,pyramidal layer of hippocampus,amygdaloid nuclei.cingulate cortex,piriform cortex and periamygdaloid cortex.The moderately dense signal was present in several thalamic and hypothalamic nuclei such as ventromedial nucleus,ventrolateral nucleus,laterodorsal thalamic nucleus,paraventricular nucleus,etc. Conclusion\ There was relationship between the gene expression of aromatase with the development of brain,there was good agreement between the distribution of aromatase mRNA and aromatase activity as previously reported.The high levels of aromatase mRNA in the region of hippocampus and cerebral cortex suggested that aromatase may implicate for sex dimorphism in cognition as well as learning and memory.\;
2.STUDY ON GENE EXPRESSION AND REGULATION OF AROMATASE IN MALIGNANT GLIOMA CELL LINE SHG-44
Acta Anatomica Sinica 2002;0(05):-
Objective To explore the gene expression of aromatase and estrogen receptor (ER-?) in malignant glioma cell line SHG-44. Methods Cell culture, immunocytochemistry, in situ hybridization and RT-PCR techniques were used. Results Aromatase and estrogen receptor gene expressions were detected in SHG-44 cells.The aromatase gene in these cells was expressed by means of the multi promoters (1^3, 1^4 and P Ⅱ).Conclusion It may provide some new data for the hormone regulation in carcinoma of nerve system.
3.EXPRESSION OF AROMATASE CYCOCHROME P - 450 IN THE NEURAL STEM CELLS AND THE EARLY DIFFERENTIATED CELLS
Lan XIAO ; Wei LI ; Wenqin CAI ; Xiangdong ZHOU
Acta Anatomica Sinica 1989;0(S1):-
Objective To explore the gene expression in the neural stem cells as well as the cells after the clone was differentiated. Methods Neural stem cell culture, Immunocytochemistry and RT-PCR technique were used. Results There was Aromatase expression in the neural stem cells, after the stem cells were differentiated, the Aromatase strongly expressed in the neruons and weakly expressed in the astrocytes. The aromatase gene in these cells was expressed by means of the brain-specific promoter 1. 4.Conclusion It may provid a new clue for the source of estrogen in the central nerve system.
6.Effects of 17?-estrogen, testosterone, tamoxifen, and aminoglutethimide on the growth and proliferation of lung adenocarcinoma cell line A549
Xiangdong ZHOU ; Wenqin CAI ; Lan XIAO ; Jiqiang ZHANG ; Zhong YANG ;
Journal of Third Military Medical University 1984;0(02):-
Objective To determine the aromatase protein expression in A549 cell and to investigate the effects of 17? estrogen (E 2), testosterone (T), estrogen receptor antagonist tamoxifen (TAM), and aromatase inhibitor DL aminoglutethimide (AMIN) on the growth and proliferation of A549 cells. Methods The expression of aromatase protein was determined by immunohistochemical methods. The changes of cell cycle and cell number before and after treatment with E 2, T, TAM, and AMIN were measured by flow cytometry and tetrazolium method (MTT). Results The aromatase protein was positively expressed in A549 cells. The aromatase inhibitor AMIN and 5?10 -7 mol/L TAM could inhibit the growth of A549 cells and block them in G 0/G 1 phase ( P
7.Knowledge and attitude of clinicians to pediatric obstructive sleep apnea hypopnea syndrome.
Feng-lei XU ; Rui-lan ZHANG ; Xiao-lan CAI ; Wei WEI
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2011;46(8):632-637
OBJECTIVETo investigate the knowledge and attitude of clinicians in the departments of pediatrics and otolaryngology to pediatric obstructive sleep apnea hypopnea syndrome (OSAHS), since in China, the clinicians in these two departments had closest relationship with the diagnosis and treatment of OSAHS in children.
METHODSA validated questionnaire from USA which was the obstructive sleep apnea knowledge and attitudes questionnaire in children (OSAKA-KIDS) was used and permission by original author. The questionnaire was mailed to ENT doctors and pediatricians in 43 public hospitals in Shandong province.
RESULTSOSA-KIDS in Chinese version was re-tested by 30 physicians, r = 0.92. Totally, 391 valid questionnaires (87.7%) were returned. Average of correct rate (x(-) ± s) in 18 knowledge items was 64.1% ± 19.1%. Cronbach's α coefficient was 0.76. There was no difference between ENT doctors and pediatrics in total knowledge score. However, there was significant difference in below 2 questions: ENT doctors had more correction in answer "nearly 2% of children have OSAHS" and pediatrics had more correction in answer "pediatric OSAHS may be associated with pulmonary hypertension". Only 24.3% clinicians correctly know the degree of snoring (mild to severe) was not correlated with the severity of obstructive apnea in children. Only 16.1% could correctly answer the question about cardio-respiratory monitor could not reliably detect both central and obstructive apnea in infant. Cronbach's α coefficient was 0.72 in 5 items which was about importance of disease and self-evaluation in confidence. While more than 90% clinicians stated that "As a clinical disorder OSAHS is important or very, extremely important". However, among them, only about 36% felt confident in identifying or managing children with OSAHS. Total knowledge score about OSAHS was not different by gender or specialty (P > 0.05), but more knowledge was associated with more positive attitudes overall (P < 0.05) and more elder in age or longer years in practice (r = 0.384, P < 0.0001).
CONCLUSIONSIt should be paid more effort to elevate the knowledge and attitude about pediatric OSAHS in pediatricians and otolaryngologists.
Adult ; Aged ; Child ; Female ; Health Knowledge, Attitudes, Practice ; Humans ; Male ; Middle Aged ; Pediatrics ; Physicians ; Sleep Apnea, Obstructive ; diagnosis ; therapy ; Surveys and Questionnaires
8.Expression and biological function of N-myc down-regulated gene 1 in human cervical cancer.
Jing, WANG ; Jing, CAI ; Zhimin LI ; Sha, HU ; Lili, YU ; Lan, XIAO ; Zehua, WANG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2010;30(6):771-6
The expression of N-myc down-regulated gene 1 (NDRG1) has previously been reported to be involved in the proliferation, differentiation, invasion and metastasis of cancer cells, but its role in cervical cancer is still unclear. This study aimed to investigate the expression of NDRG1gene in human cervical cancer and its effect on aggressive tumor behaviors. The NDRG1 expression in cervical tissues and cells was detected by RT-PCR. Specific expression plasmid pEGFP-N1-NDRG1-GFP was used to enhance the expression of NDRG1 in human cervical cancer cell lines. The mRNA and protein level of NDRG1 was assessed by RT-PCR and Western blotting, respectively. Its effects on cell proliferation, migration, invasion, cell cycle and apoptosis were detected by MTT, transwell migration assay and flow cytometry (FCM), respectively. The results showed that the expression of NDRG1 in cervical cancer tissues and cells was significantly lower than in normal cervical tissues (P<0.001). After transfection with pEGFP-N1-NDRG1-GFP, the mRNA and protein expression of NDRG1 was up-regulated in Siha cells, which suppressed cell proliferation (P<0.001), induced cell cycle arrest (P<0.05), reduced invasion and migration of Siha cells (P<0.05), but caused no cell apoptosis. Moreover, vascular endothelial growth factor (VEGF), a tumor-induced angiogenesis factor, was markedly reduced and E-cadherin, a cell adhesion molecule, was increased in the cells transfected with pEGFP-N1-NDRG1-GFP. It was concluded that up-regulated NDRG1 may play a role in the suppression of malignant cell growth, invasion and metastasis of human cervical cancer.
9.Inhibitory effect of platelet-derived growth factor-α receptor silencing on the proliferation of human lens epithelial cell
Xiao-hui, LIU ; Yan-yi, PENG ; Cai-wen, FAN ; Lan-zhen, HUANG
Chinese Journal of Experimental Ophthalmology 2013;31(8):749-753
Background Platelet-derived growth facto(PDGF) affectthe proliferation of human lenepithelial cell(LECs),and human LECexpresPDGF-α recepto(PDGFR-α) throughoutheilifetime.The binding of activated PDGF-α receptowith PDGF promotethe synthesiof DNA.Othestudiedemonstrated thasilencing of PDGFR-α by antisense oligodeoxynucleotide(ASODN) inhibitthe growth of RPE cellin proliferative vitreoretinopathy (PVR),buwhethethitechnique ifeasible foLECiunclear.Objective Thistudy wato investigate the effecof the knockdown of the PDGFR-α on the proliferation of human LECin vitro,and to offean experimental basifothe gene therapy of posteriocapsule opacification.MethodHuman LECstrain SRA01/ 04 wacultured in α-MEM containing fetal bovine serum.The cellwere incubated in 6-well platea5 × 104 cells/ well and transfection of ASODN-containing liposome waperformed.The cellwere divided into the blank control group (with blank liposome),PDGFR-α missense oligodeoxynucleotide(MSODN) group (with PDGFR-α MSODN + liposome),0.5 μmol/L PDGFR-α ASODN group (with 0.5 μmol/L PDGFR-α ASODN+liposome) and 1.0 μmol/L PDGFR-α ASODN group (with 1.0 μ mol/L PDGFR-α ASODN+liposome).The morphology of LECwaexamined undean inverse microscope 24 houraftetransfection.The expression of PDGFR-α mRNin the cellwadetected by reverse transcription-PC(RT-PCR).The rate of proliferation (A490) of the cellwaassayed using Mtand the inhibitory rate of PDGFR-α ASODN on proliferation wameasured.The percentage of LECin G1 phase waanalyzed by flow cytometer.ResultThe LECgrew well and exhibited polygonal shape in the blank control group and PDGFR-α MSODN group 24 houraftetransfection.Buin the 0.5 μmol/L and 1.0 μmol/L PDGFR-α ASODN groups,the cellappeared round in shape and the numberof cellwere obviously decreased.The expression of PDGFR-α mRNdetected by RT-Pcdemonstrated highelevel in the blank control group and PDGFR-α MSODN group;however,the PDGFR-α mRNexpression waobviously lowein the 0.5 μmol/L and 1.0 μmol/L PDGFR-α ASODN groups.The A490 value wa0.661 ± 0.036,0.655 ± 0.016,0.529 ± 0.030 and 0.441 ± 0.039 in the blank control group,PDGFR-α MSODN group,0.5 μmol/L PDGFR-α ASODN group and 1.0 μmol/L PDGFR-α ASODN group,respectively,showing significandecline in the 0.5 μmol/L PDGFR-α ASODN group and 1.0 μ mol/L PDGFR-α ASODN group in comparison with the blank control group (F=34.08,P<0.01).The percentageof LECin G1 phase were (47.73±1.18)%,(49.48±1.09)%,(53.31±1.30)% and (59.98±0.95) % in the blank control group,PDGFR-α MSODN group,0.5 μmol/L PDGFR-α ASODN group and 1.0 μmol/L PDGFR-α ASODN group,showing significandifference among them (F =68.41,P<0.01),and thain the 0.5 μmol/L PDGFR-α ASODN group o1.0 μmol/L PDGFR-α ASODN group showed significantly increase in comparison with the blank control group (P<0.05).ConclusionPDGFR-α silencing could inhibithe proliferation of human LECin vitro.
10.Expression of microRNA 27a and its correlation with drug resistance in human ovarian cancer A2780/Taxol cells
Zhimin LI ; Sha HU ; Lan XIAO ; Jing WANG ; Jing CAI ; Lili YU ; Zehua WANG
Chinese Journal of Obstetrics and Gynecology 2010;45(5):372-375
Objective To investigate the expression of microRNA 27a (miR-27a) and relationship with drug resistance in human ovarian cancer A2780/Taxol cells.Methods A stem-loop-mediated real-time PCR was used to detect miR-27a expression in A2780 and A2780/Taxol cells.The cells were transfected with the mimics or inhibitors of miR-27a or negative control RNA ( NC) by lipofectamine 2000.The expressions of MDR1 gene,P-glycoprotein (P-gp) and homeodomain-interacting protein kinase 2 (HIPK2) protein levels were measured by real-time PCR and western blot respectively.Methyl thiazolyl tetrazolium (MTT) assay was used to analyze drug sensitivity.Apoptosis analysis was measured by fluorescence activated cell sorter ( FACS).Results (1) miR-27a was an average of 2.2-fold higher expression level in A2780/Taxol cells than that in A2780 cells,with a significant difference between the two groups (P <0.05).(2) A2780/Taxol cells transfection with inhibitors of miR-27a showed that the levels of MDR1 mRNA was decreased by 39%,P-gp protein level[(26 ±5)%]decreased than that in the NC group[(43 ±7)%],HIPK2 protein level[(30 ±6)%]increased than that in the NC group[(19 ±4)%],the 50% inhibitionconcentration (0.5 (μmol/L) was less than that in the NC group (6.8 μmol/L),apoptosis rate[(32.5 ± 3.6) %]was higher than that in the NC group[(5.6 ±2.1) %],and there were significant differences between two groups (all P < 0.05 ).( 3 ) Transfection of A2780 cells with mimics of miR-27a led to increase MDR1 mRNA expression by 121% as compared with one transfection with NC (P<0.05).Conclusion The expression of miR-27a is upregulated in A2780/Taxol cells,which may regulate MDR1 and P-gp expression by targeting HIPK2.