1.Reversal of multidrug resistance in leukemic cell line K562/AO2 by chlordelazine in vitro.
Li-jun CHEN ; Shao-hua SHEN ; Hong-mei WANG ; Xin YE ; Sha-yi JIANG ; Fei GAO ; Gui-mei LI
Chinese Journal of Pediatrics 2003;41(7):525-527
OBJECTIVESome recent studies revealed that phenthiazine might be able to reverse tumor cell drug-resistance. Chlorderazin belongs to the phenthiazine compounds. The study aimed to investigate the reversing effect and mechanism of chlorderazin on multidrug resistance of leukemic cell line K562/AO2.
METHODS(1) The cytotoxicities of chlorderazin were assayed with the tetrazolium dye, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method. (2) The reverse effect of chlorderazin on K562/AO2 cells was analyzed with MTT method. The multidrug resistance reversal index (RI) was equal to the ratio of control group IC(50)/test group half inhibition concentration IC(50). (3) The intracellular daunorubicin (DNR) concentrations were measured by the flow cytometry. (4) Mdr1 mRNA expression was detected by reverse transcription-polymerase chain reaction (RT-PCR). The ratio of mdr-1/beta-actin density was calculated.
RESULTS(1) Chlorderazin 3 micro g/ml showed little toxicity to K562/AO2 cells and the suppression rate was less than 5%, so the concentration of 3 micro g/ml chlorderazin was selected as the experiment concentration. (2) The cytotoxicities of DNR to K562/AO2 were enhanced by 3 micro g/ml of chlorderazin (P < 0.05) and RI was 1.901. (3) Chlorderazin of 3 micro g/ml could increase the intracellular DNR accumulation significantly (P < 0.05), and the fluorescence staining by the flow cytometry was higher (250.95 +/- 18.96) than the control group (112.75 +/- 15.78) and shift right in K562/AO2 cells treated with chlorderazin, and the difference was significant (P < 0.05). (4) Chlorderazin has no significant influence to the expression level of mdr-1 mRNA. Both test group and control group showed a clear mdr-1 mRNA band located at the position of 157 kb. The ratios of mdr-1/beta-actin density were 0.414 +/- 0.012 in the test group and 0.447 +/- 0.027 in the control group, respectively, and the difference was not significant (P > 0.05).
CONCLUSIONChlorderazin could reverse the multidrug resistance by increasing the intracellular DNR accumulation in K562/AO2 cells. The effects had no correlation to the mdr-1 gene. Further study is needed.
ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; Antiemetics ; pharmacology ; Cell Division ; drug effects ; Chlorpromazine ; pharmacology ; Drug Resistance, Multiple ; drug effects ; genetics ; Drug Resistance, Neoplasm ; drug effects ; Flow Cytometry ; Humans ; K562 Cells ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction
2.The analysis of allelic deletion in 4 cases of Amelogenin loci in paternity testing
Yi-Dong LIU ; Yi-Kai HU ; Sha LI ; Hong-Mei XU
Fudan University Journal of Medical Sciences 2017;44(z1):21-25
Objective To apply PowerPlex(R) 21 System Kit and AGCU 21 + 1 STR Fluorescence Detection Kit as supplementary detection kit in some samples with Amelogenin locus X deletion.Methods While with the help of Investigator Argus X-12 Kit,MicroreaderTM 19X ID System kit and the application of capillary electrophoresis,the complete X pattern of these four samples can be identified.Results From Mar,2013 to Dec,2016,4 cases of male samples with Amelogenin locus X deletion have been found.Conclusions Sometimes Amelogenin locus X deletion may happen when using PowerPlex(R)21 System Kit and AGCU 21 + 1 STR Fluorescence Detection Kit,while validating with other forensic biology detection kit can assure the accuracy of genotyping.
3.Effects of heme oxygenase-1 on proteins related to apoptosis in INS-1 cells exposed to intermittent high glucose.
Yi-guang CHEN ; Yao-ming XUE ; Mei-ping GUAN ; Bo ZHU ; Jian-ping SHA
Journal of Southern Medical University 2010;30(10):2330-2332
OBJECTIVETo study the effect of heme oxygenase-1(HO-1) on proteins related to apoptosis in INS-1 cells with exposure to intermittent high glucose.
METHODSINS-1 cells cultured in vitro were divided into control group, persistent high glucose group (PHG), intermittent high glucose group (IHG), CoPP + intermittent high glucose group (CoPP+IHG), and ZnPP+ intermittent high glucose group (ZnPP+IHG). After 72 h of treatment with the corresponding protocols, the cells were examined for expressions of HO-1 protein by Western blotting and for expressions of Bax and Bcl-2 by immunocytochemistry.
RESULTSIn comparison with the control group, the cells in both PHG group and IHG group showed significantly increased expressions of HO-1 (P<0.01) and decreased Bcl-2/Bax ratios (P<0.05). The cells in CoPP+ IHG group exhibited a greater HO-1 protein expression but a lower Bcl-2/Bax ratio than those in IHG group (P<0.05) The ZnPP+IHG group demonstrated opposite changes in terms of HO-1, Bax and Bcl-2 expressions compared with the CoPP+IHG group.
CONCLUSIONIntermittent high glucose can lower Bcl-2/Bax ratio in INS-1 cells, and HO-1 may protect INS-1 cells against apoptosis possibly by up-regulating the Bcl-2/Bax ratio.
Animals ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cell Line ; Glucose ; administration & dosage ; adverse effects ; Heme Oxygenase (Decyclizing) ; metabolism ; Islets of Langerhans ; cytology ; drug effects ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; bcl-2-Associated X Protein ; metabolism
4.Investigation of nutrition and feeding status among children under 2 years in Beijing.
Yao ZHAO ; Lei HUANG ; Zheng ZHANG ; Xing-huo PANG ; Yi-mei SHA ; Zhong DONG
Chinese Journal of Preventive Medicine 2006;40(1):46-49
OBJECTIVETo investigate the nutrition status, breast feeding and complementary feeding among children under 2 years of age in Beijing.
METHODSA stratified cluster randomly sampling method was used. After having got the compliance a questionnaire survey was carried out to 422 children < 2 year-old in total 18 districts of Beijing.
RESULTSThe average infant birth weight was 3325 g, the rate of weight (2500 g and > or = 4000 g were 2.8% and 13.0% respectively. The rate of exclusive breast-feeding, partial breast-feeding, bottle-feeding of infant under 4 months was 56.6%, 33.9% and 9.5% respectively. The average weaning time was 7.4 months. The rate of complementary feeding of infant up to 6 months was 99.4%. The average ages for addition of complementary feeding such as cereal, vegetable and fruit, egg, oil, nutrients in rural areas were later than those in urban.
CONCLUSIONIt is important to promote breast feeding and improve complementary feeding by education and intervention.
Bottle Feeding ; statistics & numerical data ; Breast Feeding ; statistics & numerical data ; China ; Humans ; Infant ; Infant Food ; statistics & numerical data ; Infant, Newborn ; Nutrition Surveys ; Rural Population ; statistics & numerical data ; Surveys and Questionnaires ; Urban Population ; statistics & numerical data
5.Preparation, characterization of paclitaxel-loaded Pluronic P105 polymeric micelles and in vitro reversal of multidrug resistant tumor.
Yong-Zhong WANG ; Xiao-Ling FANG ; Ya-Juan LI ; Zhi-Wen ZHANG ; Li-Mei HAN ; Xian-Yi SHA
Acta Pharmaceutica Sinica 2008;43(6):640-646
Drug delivery system (DDS) is a novel approach to overcome multidrug resistance (MDR) in tumors nowadays. This work was designed to investigate a new micellar delivery system for in vitro reversal of resistant ovarian tumor cells, based on a nonionic triblock copolymer Pluronic P105 and paclitaxel (PTX). The PTX-loaded polymeric micelles (P105/PTX) were prepared by thin film-hydration methods. Based on the results of single factor experiments, the P105/PTX micelle formulation was optimized by employing the central composite design-response surface methodology. The physico-chemical properties of the P105/PTX micelles were characterized, including micelle size, drug loading coefficient, in vitro release behavior, etc. The cytotoxicity of the P105/PTX micelles was assessed against human ovarian tumor cell line, SKOV-3/PTX, by a standard 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl (MTT) assay. In order to understand the possible mechanism of Pluronic effects in resistant tumor cells, cellular uptake study of micellar PTX or Rhodamine-123 (R-123) was also carried out. The results showed that the micelle size was about 24 nm with drug loading coefficient of 1.1% and PTX concentration of 700 microg x mL(-1). The cumulative release amount of PTX from the P105/PTX micelles was only 45.4% in 6 h (P < 0.05) and 79.6% in 24 h, whereas Taxol injection in 6 h released 95.2% PTX. The IC50 values of the P105/PTX micelles and Taxol injection against SKOV-3/PTX were 1.14 and 5.11 microg x mL(-1), and resistance reversion index (RRI) was 9.65 and 2.15, respectively. The micellar PTX or R-123 exhibited a significant increase in cellular uptake in resistant SKOV-3/PTX cells compared with free PTX or R-123. These results indicated that PTX could effectively be solubilized by Pluronic P105 block copolymers via thin film-hydration process and formulation optimization, producing nano-scale polymeric micelles with sustained release property in vitro. The P105/PTX micelles were effectively able to reverse resistance to PTX in SKOV-3/PTX tumor cells compared with Taxol injection or free PTX solution, and the enhanced cytotoxicity in the resistant SKOV-3/PTX cell was related to the improved cellular uptake of PTX by Pluronic P105 copolymers.
Antineoplastic Agents, Phytogenic
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administration & dosage
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chemistry
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pharmacology
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Cell Line, Tumor
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Drug Carriers
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Drug Delivery Systems
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Drug Resistance, Multiple
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drug effects
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Drug Resistance, Neoplasm
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drug effects
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Excipients
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chemistry
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Female
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Humans
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Inhibitory Concentration 50
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Micelles
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Ovarian Neoplasms
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metabolism
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pathology
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Paclitaxel
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administration & dosage
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chemistry
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metabolism
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pharmacology
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Particle Size
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Poloxamer
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chemistry
6.Study of the HLA-A, B, DRB1 allele and haplotype frequencies of 4194 cord blood samples in Guangzhou.
Wen-qiong SHA ; Can LIAO ; Yan LI ; Xin YANG ; Xue-wei TANG ; Yi-ning HUANG ; Xing-mei XIE
Chinese Journal of Medical Genetics 2008;25(4):473-476
OBJECTIVETo analyze the distribution of the human leukocyte antigen (HLA)-A, B and DRB1 allele and haplotype in cord blood samples preserved in Guangzhou Cord Blood Bank collected in the last 10 years.
METHODSThe HLA-A, B and DRB1 genotyping of 4194 cord blood samples were detected by Special Monoclonal Tray, PCR-sequence specific promer (PCR-SSP), PCR-sequence specific oligonucleotide probe (PCR-SSO) and sequence based typing (SBT). Frequencies of HLA-A, B and DRB1 allele and haplotype were calculated by Arlequin software.
RESULTSThe total numbers of HLA-A, B and DRB1 alleles are 18, 43, 13 respectively. The obviously high frequency alleles are A*11, A*02, A*24, A*33, B*40, B*15, B*46, B*13, DRB1*12, DRB1*15, DRB1*09 and DRB1*04, with accumulative frequency of each locus being more than 50%. The most common haplotypes are A2-B46, B46-DR9, A11-DR12 and A2-B46-DR9.
CONCLUSIONThe distribution of HLA-A, B and DRB1 allele and haplotype of cord blood in Guangzhou Cord Blood Bank has typical characteristics of southern Chinese Han population. Authors' data may help in searching for appropriate donors.
Alleles ; Asian Continental Ancestry Group ; genetics ; China ; Female ; Gene Frequency ; Genetics, Population ; HLA-A Antigens ; genetics ; HLA-B Antigens ; genetics ; HLA-DR Antigens ; genetics ; Haplotypes ; Humans
7.Analysis on influence factors of dietary supplement used in population aged above 45 years in Beijing.
Yu-na HE ; Zhen YANG ; Jun XU ; Yi-mei SHA ; Zhi-yong REN ; Xing-huo PANG ; Guo ZENG ; Feng-ying ZHAI
Chinese Journal of Preventive Medicine 2008;42(11):823-826
OBJECTIVETo explore influence factors of dietary supplement used in population aged above 45 years in Beijing.
METHODSUtilizing the data of the survey of dietary supplement (DS) used in Beijing in 2006 was to investigate the influence factors by using multi-factorial logistic regression analysis.
RESULTSAll 2487 samples were included in the analysis. Sex, age, average income of each month for each member of the family, experiences of deficiency of nutrition, attitude to nutrition knowledge, attitude to the need of DS and city-or-rural resident had shown effects on DS using. The percentage of using DS in female was higher than that in male being 19.3% and 11.8% respectively. The percentage grew up as the age became older. The same trend appeared as the economic-status going up, but not as education level change. Use of DS was greater in urban population (16.9%) than in rural population (14.6%). The percentage of using DS in people who had or just been diagnosed as malnutrition/deficiency disease or chronic disease, or those who thought themselves having got these diseases only by their own feeling were higher than those did not.
CONCLUSIONPeople the female, the elder, or those having higher income, having experiences of deficiency of nutrition, are more interested in nutrition knowledge, and more positive in need of DS and those living in rural should be the target population of nutrition education for DS use.
Age Factors ; Aged ; Aged, 80 and over ; China ; Dietary Supplements ; statistics & numerical data ; Female ; Humans ; Male ; Middle Aged ; Nutrition Surveys ; Sex Factors
8.Protection of MICU1 against myocardial hypertrophy induced by angiotensin Ⅱ
Yi YANG ; Yue TIAN ; Yang SHEN ; Mei-Li GUAN ; Sha CHEN ; Jian FENG ; Hai-Feng PEI ; Yong-Jian YANG
Medical Journal of Chinese People's Liberation Army 2017;42(12):1051-1055
Objective To investigate the role of mitochondrial calcium uptake 1 (MICUI) in myocardial hypertrophy of mice and underlying mechanism.Methods The model of myocardial hypertrophy was established via incubation of mouse cardiac myocytes (MCM) with 300nmol/L angiotensin Ⅱ (Ang Ⅱ) for 48 hours in vitro.After that,MICU1 specific small interfering RNA (siRNA) was delivered to knockdown MICU1 levels in MCM.On the other hand,adenovirus-mediated over-expression of MICU 1 was transfected into MCM.Accordingly,the expressions of ANP and BNP in myocardial cells were measured by qRT-PCR.Mitochondrial membrane potential and ATP contents were detected byJC-1 assay kit and ATP assay kit,respectively.Then,Western blotting and qRT-PCR were used to detect the levels of MICU1 in myocardial cells.The mitochondrial Ca2+ contents were measured via atomic absorption flame spectroscopy.The size of myocardial cells was determined by α-actinin staining.Results Mitochondrial membrane potential and ATP contents in hypertrophic cardiomyocytes induced by Ang Ⅱ were both decreased.Meanwhile,myocardial hypertrophy significantly increased mitochondrial Ca2+ contents but decreased MICU1 levels.With the method of genetic intervention,we found that MICUI deficiency exacerbated mitochondrial Ca2+ overload,increased cell surface and elevated the expression of BNP.Conversely,the overexpression of MICU1 obviously decreased mitochondrial Ca2+ overload,cell surface of MCM and expressions of ANP and BNP.Conclusion MICU1 alleviates Ang Ⅱ-induced myocardial hypertrophy via inhibiting mitochondrial Ca2+ overload.
9.Pregnancy outcomes among women with high and low risk in first and second trimester prenatal screening analysis
Yi-Ming CHEN ; Wen ZHANG ; Sha LU ; Yan-Zhen ZHANG ; Jie-Jing LIAN ; Li-Dan ZHANG ; Min WANG ; Hao WANG ; Jin MEI ; Wen-Sheng HU
Journal of Preventive Medicine 2018;30(3):240-243,247
Objective To analyze adverse pregnancy outcomes among women with high risk and low risk during prenatal screening. Methods Clinical data of 180006 pregnancies in 5 prenatal screening center in Hangzhou were collected. We compared the adverse pregnancy outcomes of high and low risk pregnancies. Results Among 180006 pregnancies (age<35 years old), there were 10296 high-risk cases and 169710 low-risk cases, with 168654 cases followed. There were 9406 high-risk cases of Down's syndrome (DS) (5.23%), 273 high-risk cases of Edwards' syndrome (ES) (0.15%) and 617 high-risk cases of open neural tube defect (ONTD) (0.34%) . The detection rate of pregnancy outcome of premature birth, spontaneous abortion, termination of pregnancy, stillbirth in the high risk was 5.46%, 0.80%, 1.80%, 0.37%, respectively, and that rate in low risk results was 3.50%, 0.21%, 0.38%, 0.18%, respectively. Adverse pregnancy outcomes in high-risk group were significantly higher than that in low-risk group (all P<0.001) . The actual incidence rate of DS, ES or ONTD in high-risk group (4.56‰, 1.65‰, 0.97‰, respectively) were significantly higher than that in low-risk group (0.12‰, 0.04‰, 0.09‰, respectively, all P<0.001) . The detection rates of prenatal screening were 70.15%, 68.00% and 38.46%, and false positive rates were 5.23% , 0.14% and 0.34% . Conclusion High risk of serum prenatal screening analysis is associated with adverse pregnancy outcomes. The number of pregnancies experienced invasive prenatal diagnosis can be reduced by routine serum prenatal screening. That is an Effective method to reduce the birth defects.
10.Expression and significance of microRNA-134 in mouse brain tissue with FMR1 gene knockout
Zhi-Yong ZENG ; Wei DI ; Du XIAO ; Xun-Sha SUN ; Yu-Liang WANG ; Mei OU-YANG ; Yong-Hong YI
Chinese Journal of Neuromedicine 2010;09(8):765-767
Objective To observe the expression ofmicroRNA-134 (miR-134) in the mouse brain tissue with FMR1 gene knockout during the different development periods and its expression characteristic, and explore whether the deficiency of fragile X mental retardation protein (FMRP) can induce the changes of miR-134 transcription. Methods FVB strain male mice, including FMR1 gene knockout (KO, n=15) and their wild type (WT, n=15) counterparts were chosen in the experiment. The expressions of miR-134 in the brain tissues of these KO mice that were 0 d, 4 and 6 w old and the age-matched WT mice were detected by qRT-PCR. Results The transcriptional level of miR-134 in the brain tissue of KO mice had no significant difference as compared with that of age-matched WT mice (P>0.05). The transcriptional levels of miR-134 in 6-w-old KO and WT mice were significantly decreased as compared with the newbom and 4-w-old same genotype mice (P<0.05). Conclusion The absence of FMRP does not influence the transcription of miR-134 and the transcriptional level of miR-134 in the brain tissues maintains a high level during the developmental stage of the nervous system and gradually decreases to a low level after grow-up, demonstrating its important role in regulating the development of nervous system.