1.Molecular mechanism of indirubin-3'-monoxime and Matrine in the reversal of paclitaxel resistance in NCI-H520/TAX25 cell line.
Su-xia LUO ; Wen-ying DENG ; Xin-feng WANG ; Hui-fang LÜ ; Li-li HAN ; Bei-bei CHEN ; Xiao-bing CHEN ; Ning LI
Chinese Medical Journal 2013;126(5):925-929
BACKGROUNDMultidrug resistance (MDR) is a main reason for paclitaxel (TAX) treatment failure. Indirubin-3'-monoxime (IRO) and Matrine are traditional Chinese medicines, which may reverse the resistance of tumor cells to some chemotherapy drugs, but the relationship between paclitaxel resistance and Matrine is still unclear. The aim of this study was to explore the potential molecular mechanism of IRO and Matrine in reversal of TAX resistance.
METHODSIn this study, MTT assay was used to measure the non-cytotoxic dosage of IRO and Matrine on NCI-H520/TAX25 cells and determine the reversal extent of TAX resistance under non-toxic doses. In addition, RT-PCR and Western blotting were used to evaluate the mRNA expression and the protein level of survivin, Oct-4, and Sox-2 in NCI-H520/TAX25 cells using semi-quantitative methods.
RESULTSThere was no obvious inhibition on sensitive cell strains and drug-resistant strains, when the final concentration was at lest 4 µmol/L for IRO and 100 µmol/L for Matrine. So 4 µmol/L of IRO and 100 µmol/L of Matrine were considered as the reversal dosage. When 4 µmol/L of IRO or 100 µmol/L of Matrine were used together with TAX, the sensitivity to TAX increased evidently in NCI-H520/TAX2 cells; the reversal rate of IRO and Matrine was about 1.92 (43.56/22.6 nmol/L) and 1.74 (43.56/25.0 nmol/L), respectively. The mRNA expression and the protein level of survivin, Oct-4, and Sox-2 in NCI-H520/TAX25 decreased significantly (P < 0.05) after addition of IRO or Matrine in TAX treatment, compared to that of TAX treatment alone.
CONCLUSIONThe decrease in both mRNA expression and protein level of survivin, Oct-4, and Sox-2 might be the molecular mechanism, by which IRO and Matrine mediate the reversal of TAX resistance.
Alkaloids ; pharmacology ; Blotting, Western ; Cell Line, Tumor ; Drug Resistance, Neoplasm ; Humans ; Indoles ; pharmacology ; Inhibitor of Apoptosis Proteins ; genetics ; metabolism ; Octamer Transcription Factor-3 ; genetics ; metabolism ; Oximes ; pharmacology ; Paclitaxel ; pharmacology ; Quinolizines ; pharmacology ; SOXB1 Transcription Factors ; genetics ; metabolism
2.Prokaryotic expression, purification of prM of JEV and preparation of monoclonal antibody.
Bei-fang NING ; Huai-min ZHU ; Xiao-jun ZHOU ; Yi CAO ; Ai-guo ZHOU
Chinese Journal of Experimental and Clinical Virology 2008;22(1):65-67
OBJECTIVETo prepare monoclonal antibody (mAb) against prM epitope.
METHODSThe gene encoding prM was isolated using RT-PCR from brain of JEV infected mouse and cloned into prokaryotic expression vector pET-32a. Recombinant plasmid was transformed into E.coli BL21/DE3/LysS, then the transformed cells were expressed with the induction of IPTG. The expression and purification of the prM protein was analyzed by SDS-PAGE. The BALB/c mice were immunized with purified prM protein. Hybridoma cell lines secreting monoclonal antibodies against prM were established after cell fusion of mouse splenic cell and P3-X63-Ag8.653 cells. The specificity of mAb was identified by ELISA, Western Blot and Immunohistochemistry assay.
RESULTSmAb against prM epitope of JEV was prepared successfully.
CONCLUSIONThe obtained prM specific mAb was valuable for the prevention and dignosis of Japanese encephalitis.
Animals ; Antibodies, Monoclonal ; analysis ; immunology ; isolation & purification ; Antibody Specificity ; BALB 3T3 Cells ; Cell Line ; Cloning, Molecular ; Electrophoresis, Polyacrylamide Gel ; Encephalitis Virus, Japanese ; genetics ; immunology ; Epitopes ; immunology ; Escherichia coli ; genetics ; Mice ; Plasmids ; genetics ; metabolism ; Prokaryotic Cells ; metabolism ; Sequence Analysis, DNA ; Viral Proteins ; biosynthesis ; genetics ; immunology ; isolation & purification
3.Multi-detector-row CT findings of ACTH-independent macronodular adrenal hyperplasia
Qi SONG ; Weiwei ZHOU ; Tingwei SU ; Lianjun DU ; Bei DING ; Huan HANG ; Zilai PAN ; Kemin CHEN ; Fuhua YAN ; Weiqing WANG ; Guang NING ; Wenqiang FANG
Chinese Journal of Endocrinology and Metabolism 2012;28(4):335-339
ObjectiveTo explore the characteristics multi-detector-row computed tomography (MDCT)findings of ACTH-independent macronodular adrenal hyperplasia ( AIMAH ).Methods The un-enhanced and contrast-enhanced MDCT features in 24 patients ( 14 males and 10 females) with clinically confirmed AIMAH were retrospectively assessed for the morphology and enhancement patterns.ResultsThe adrenal glands were involved bilaterally in all of the 24 cases( 100% ).24 patients had massively enlarged multinodular adrenal glands.Nodules were( 1.79 ± 1.02) cm (0.50 ~ 3.85 cm),which usually distorted and completely obscured the normal adrenal glands.The enlarged adrenal glands were still retained the adreniform contour,showed characteristic ginger-like.22 of the 24 ( 91.7 % ) hyperplastic nodular glands demonstrated mild homogeneous enhancement.Calcification was revealed in 1 adrenals ( 1/24,4.2% ).Conclusion MDCT reveals the characteristic morphology and CT attenuation in AIMAH.Combined with its clinical presentation and biochemical findings,AIMAH is able to be diagnosed with high specificity and accuracy on MDCT.
4.Study on the comparison of high risk behaviors related to AIDS between heterosexual and homosexual men among men who have had sex with men.
Liu-Mei LIAO ; Bei-Chuan ZHANG ; Xiu-Fang LI ; Ming-Hua LIU ; Hui LI ; Ning WANG ; Quan-Ping AN ; Zeng-Zhao YU
Chinese Journal of Epidemiology 2007;28(9):844-847
OBJECTIVETo study the risk sexual behaviors related to AIDS between heterosexual and homosexual men who have had sex with men.
METHODSTarget sampling, anonymous questionnaires were adopted to compare sexual behaviors between the two groups.
RESULTSThe total amount of sex partners with same-sex was 13.8 on average among heterosexual men including 3.8 with male partners in the past 6 months. Numbers of male partners who had oral sex with was 10.5 and anal sex was 12.4 which were both less than with same sex. Among heterosexual men, the total number of female partners was 4.9 on average but number of female partners in the past 6 months was 1.7 which were both more than that among the homosexuals who were all in marriage status. The rate of condom use was 68.8% (lower than that among homosexual men), among heterosexual men when having sex with men. The rate of condom use among heterosexual men during last anal intercourse was 91.3% with male partners or 63.7% with females. Both figures were higher than that among the homosexuals. The rates of condom use among the two groups were 91.3% and 71.0% respectively during the last anal intercourse with men which were higher than the corresponding rate of condom use during the last oral sex. The rate of heterosexual men who ever had engaged in group sex was 9.9% in the previous year and the incidence of bleeding was 16.7% during sexual intercourse. 11.4% of them reported ever having had sex with partners from other areas in the last year and 4.2% had experienced same-sex harassment before 16 years of age. 4.6% had paid for male-male sex. All these figures were lower than that of the homosexuals.
CONCLUSIONThe characteristics of high risk sexual behaviors related to AIDS showed much difference in the two groups which called for attention among these groups of MSM.
Acquired Immunodeficiency Syndrome ; epidemiology ; Adult ; Condoms ; utilization ; Heterosexuality ; statistics & numerical data ; Homosexuality, Male ; statistics & numerical data ; Humans ; Male ; Risk Factors ; Risk-Taking ; Sexual Partners ; Young Adult
5.Associations between partner-seeking activities through internet and sexual risk behaviours related to HIV/AIDS among men who have sex with men in China
Xiu-Fang LI ; Bei-Chuan ZHANG ; Quan-Sheng CHU ; Ning WANG ; Yan-Fei WANG ; Zeng-Zhao YU
Chinese Journal of Epidemiology 2008;29(7):685-688
Objective To explore the associations between partner-seeking activities through internet and HIV/AIDS related risk behaviours among men who have sex with men(MSM).Methods Target sampling.anonymous questionnaires were adopted and analyzed with SPSS 11.0.Results Data from 45.6% of the 2178 valid questionnaires showed that most partner-seeking activities was through internet.73.1% of them landed on gay website.in which 83.0% looked for sexual partners and 91.7% of them had intercourse with other men met through internet when compared with MSM who did not seek sex activities through internet.The internet group obviously had fewer numbers of partners(5.73 vs.10.3),oral sex partners(5.13 vs.8.71),passive anus sex partners(4.67 vs.8.26)and active anus sex partners (3.51 vs.6.06)in the past 6 months(P<0.01 respectively)and obviously had fewer group sex(16.7% vs.20.5%)activities in the past one year but had more activities on oral sex(94.7% vs.89.6%),anus sex(95.1% vs.86.5%),regular sexual partner(52.6% vs.46.7%)in the past 6 months(83.4% vs.75.5%)(P<0.01 respectively)and more likely to have finger intercourse(52.2% vs.38.5%),hemorrhage sexual intercourse(36.4% vs.21.9%,OR=2.04),non-local sexual intercourse(28.1% vs.22.7%)(P<0.01 respectively).Conclusion Seeking sexual partners through internet had positive and double effects on MSM's risk behaviours.suggesting that it is of urgent need to take the advange of internet and gay website to disseminate HIV/AIDS prevention message.
6.The impact of childhood sexual abuse on the development of AIDS related high risk behaviors and psychological appearances among men who have sex with men
Zeng-Zhao YU ; Bei-Chuan ZHANG ; Xiu-Fang LI ; Ning WANG ; Tong-Xin SHI ; Quan-Sheng CHU
Chinese Journal of Epidemiology 2009;30(1):14-17
Objective To study the AIDS related high risk behaviors and psychological appearances among men who have sex with men (MSM) who ever experienced childhood sexual abuse (CSA).Methods Target sampling for a cross-sectional study was developed and valid anonymous questionnaires were adopted to compare the differences of high risk behaviors related to AIDS and psychological appearances betwen those with or without CSA experiences among 2147 MSM from nine cities.Results Compared to corresponding ones without CSA experience,CSA group had a significant larger numbers in the following events: total sexual partners,anal sex episodes with same sex,female sexual partners and anal sex in the previous six months,with the figures of median as 20.0,10.0,3.0,3.0 respectively.In the previous year,30.8% of them had ever participated in 'group sex',19.2% ever exchanged money for sex,36.7% bled while having sexual intercourse,37.3% had sex with male partners away from his own region.All the above said figures were higher than non-CSA group,with significant differences.It also appeared that CSA experience had an impact on significant lower rate of condom use (67.3%) in the last anal sex.Those with CSA experience had more psychological problems which appeared as: 75.6% considered they would suffer from serious discrimination if their sexual orientation ever disclosed,34.7% had a strong intention of suicide and 24.3% ever having had suicidal attempts.The differences of the two groups showed statistical significance.Condusion CSA experience not only increased the number of AIDS related high risk behaviors in adulthood,but also had negative impact on their psychological appearances.It is of urgent need to carry out psychological intervention approaches to target on MSM with CSA experiences while childhood sexual education and rights assurance towards juvenile population should also not be neglected.
7.Immunoregulation effects in vitro of the xenoprotein in combination with recombinant human granulocyte-macrophage colony stimulating factor and bacillus Calmette-Guerin.
Ming-Li WANG ; Zhi-Gang XIE ; Han LU ; Ming SHI ; Mei-Ru HU ; Ming YU ; Yuan-Fang MA ; Bei-Fen SHEN ; Ning GUO
Journal of Experimental Hematology 2008;16(6):1408-1412
This study was aimed to investigate the effects of xenogeneic antigen neu-Fc in combination with the recombinant human granulocyte-macrophage colony stimulating factor (GM-CSF) and Bacillus Calmette-Guerin (BCG) on the regulation of Th1 and Th2 immune response in vitro. The rat neu L2-S2 domain was engineered as a chimeric protein with human IgG Fc. The eukaryotic expression vector was constructed. The recombinant protein was stably expressed in CHO cells and purified by rProtein A Sepharose Fast Flow column. The recombinant protein was identified by SDS-PAGE and Western blot. Peripheral blood mononuclear cells (PBMNCs) were obtained by means of standard Ficoll separation from the blood of healthy donors. Neu-Fc-induced PBMNC proliferation was tested by MTT. The production of IL-12 and IL-10 was measured by ELISA. The results showed that the level of IL-12 decreased and IL-10 increased after PBMNCs were incubated with MCF-7 cultural supernatant. 10 nmol/L neu-Fc strongly induced the cell proliferation. Compared with neu-Fc or GM-CSF or BCG treatment alone, neu-Fc in combination with GM-CSF and BCG significantly stimulated IL-12 production and inhibited IL-10 production (p < 0.01). It is concluded that the neu-Fc can stimulate the proliferation activity of PBMNCs. neu-Fc, GM-CSF and BCG costimulation efficiently induces Th1 immune response.
Animals
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BCG Vaccine
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immunology
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CHO Cells
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Cricetinae
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Cricetulus
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Granulocyte-Macrophage Colony-Stimulating Factor
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immunology
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Humans
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Interleukin-10
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metabolism
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Interleukin-12
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metabolism
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Rats
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Recombinant Proteins
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immunology
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Th1 Cells
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immunology
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Th2 Cells
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immunology
8.Study on the comparison of high risk behaviors related to AIDS among different status of income in men who have had sex with men
Tong-Xin SHI ; Bei-Chuan ZHANG ; Xiu-Fang LI ; Jing-Xing XU ; Ning WANG ; Sheng-Jian ZHOU ; Wen-Hui GUAN ; Jing-Dong ZHANG
Chinese Journal of Epidemiology 2008;29(5):426-429
Objective To evaluate the effects of socioeconomic status on the distribution of cardiovascular risk factors and clinicaltreatments of patients with acute myocardial infarction in Beijing.Methods In Beijing, a prospective, muhi-center, registration study was carried out which including 800 patients who were consecutively hospitalized for ST-segment elevation acute myocardial infarction within 24 hours after event attack in 19 different hospitals in Beijing between November, 2005 and December, 2006.Indicators of socioeconomic status included self-reported personal income (<500, 500-2000,>2000 RMB/month), educational attainment (≤ 12 and > 12 years) and status of medical insurance (yes/no).According to categories of education, patients were categorized into two groups of lower socioeconomic status and higher socioeconomic status. Differences of cardiovascular risk factors and clinical treatments were compared across the two groups respectively. Results Proportion of diabetes and hyperlipidemia in patients with higher socioeconomic status was much higher than that of patients with lower socioeconomic status (P<0.05, P<0.01 respectively). Patients with lower socioeconomic status were more likely to be smokers (P <0.05). The rates of receiving coronary angiography and PTCA were much lower in patients with lower socioeconomic status. Medical insurance and income were the most important two socioeconomic factors determining the use of PTCA. Conclusion Compared to patients with lowersocioeconomic status,patients with higher socioeconomic status had higher rates of hyperlipidemia and diabetes but lower smoking rate among cardiovascular risk factors. The rates of receiving interventional therapies were much lower in patients with lower socioeconomic status.
9.Study on HIV/AIDS related high risk behaviors among unmarried men who have sex with women
Tong-Xin SHI ; Bei-Chuan ZHANG ; Xiu-Fang LI ; Jing-Xing XU ; Ning WANG ; Zeng-Zhao YU ; Yan-Fei WANG
Chinese Journal of Epidemiology 2009;30(1):10-13
Objective To explore the risk of sexual behaviors related to AIDS among unmarried men who often have sex with women.Methods Study objectives were recruited through target sampling and anonymous questionnaires were adopted.Men' s sexual behaviors were compared with each other between the group of men who always had sex with women (group A) and who never had sex with women (group B) in the past 6 months.Results The average numbers of same-gender partners among group A who acted as the male role in the past 6 months and who had anal intercourse in the last month were 5.64 and 3.09 respectively,which were both higher than those of group B's (P< 0.05).The rate of condom use among men in group A when having sex with men was 81.94% .The percentages of condom use among men in group A who having had oral or anal sex with male sex buyers last time were 15.38% and 70.27% respectively.They were all higher than those of group B' s (P<0.05 or 0.01 ).The rates of men in group A who were male sex workers and who having had sex with unknown men in gay places during the past 6 months were 27.59% and 61.70% respectively.Among group A,the percentages of men who had had sex with unknown men during the last time and who had had sex with nonlocal male partners during the past year were 43.06% and 40.28% respectively.They were all higher than those of group B's (P<0.01).The percentage of condom use among men in group A when having had sex with women during the past 6 months was 68.94%,but only 25.96% of the men used condom regularly.The rate of condom use in men from group A when having sex with women during the last sexual intercourse was 65.45%.Conclusion HIV high-risk sex behaviors were more ubiquitous among unmarried MSM who always had sex with women,suggesting that different interventions be developed to prevent them from transmitting HIV from MSM to women or other men.
10.Study on the high risk behaviors related to AIDS among men who having sex with men in the floating population
Tong-Xin SHI ; Bei-Chuan ZHANG ; Xiu-Fang LI ; Jing-Xing XU ; Ning WANG ; Zeng-Zhao YU ; Yan-Fei WANG
Chinese Journal of Epidemiology 2009;30(7):668-671
Objective To explore the risks on sexual behaviors related to AIDS among men who having sex with men (MSM) in floating population and to develop relevant interventions. Methods Study objectives were recruited through target sampling and anonymous questionnaires were adopted. Male sexual behaviors were compared between floating population and permanent registered residents.Results Among those MSM, the average numbers of same-gender partners, oral sex, anal sex and sex with females during the past 6 months among floating population were 9.61, 8.61, 7.52 and 1.17 respectively. Numbers of partners on oral sex and anal sex during the past month were 2.89 and 3.69 respectively. Both figures were higher than those of the permanent registered residents (P<0.05 or P< 0.01). The rates of condom use among floating population when having anal sex with men or with non-marital female partners were 37.15% and 23.28% respectively. The figures were all higher than those of permanent registered residents' (P<0.05 or P<0.01). Among floating population, the percentages of men who had sex with male sex workers or who had sex with men acquainted through inter-net or with stable female sex partners were 17.89%, 12.56%, and 31.86% respectively. The rates of men who had group sex among gay men, who bled when having sex with men or who had sex with men from other areas during the past year were 22.13%, 31.38% and 32.36% respectively. The figures were all higher than those of permanent registered residents' (P<0.05 or P<0.01). Conclusion Sex-related high risk behaviors were more ubiquitous among floating population than those of permanent registered residents because they were more likely to transmit HIV to people from/to other areas. More attention should be paid to the floating population with MSM in particular, when health interventions are carried out.