1.Experience exchange in vitro micronucleus assay
Chinese Journal of Comparative Medicine 2016;26(3):86-88
Objective To compare the conventional marrow collection with new marrow collection, in the number of cell, other tissues pollution and the background of the smear, providing the reference for future micronucleus test. Methods The mice was enthanasia and the sternums were taken.One group, using the conventional method of marrow collection, squeezing the marrow to the slide with fetal bovine serum;the other group, using 1-mL injector extracting fetal bovine serum 100μL, injecting into mice sternums and rushing out the bone marrow for circle smear.Results Two methods can meet the requirement of test, but the new marrow collection can acquire more number of cells, less the tissues pollution and more clear in the background of smear.Conclusions Comparing with the conventional marrow collection, the new method has more superiority to simplify the next cell counting.
2.Research progress of the methods and applications of micronucleus assay
Si CHEN ; Keqing LU ; Xingming MA
Chinese Journal of Comparative Medicine 2016;26(2):83-86
Micronucleus ( MN) assay as a routine examination for genotoxicity has been widely used.The testing specimens were taken from bone marrow and extended from blood and tissues.In addition to testing genotoxicity of drugs, it is also applied in disease diagnosis for genetic mutation, evaluation of curative effectiveness and disease prevention. Moreover, MN assay is also an important safety indicator for drugs and health foods registration.This review will discuss the staining method of MN test and its application in the field of diseases and virology.
3.Mechanism of AMPK-enhanced chemosensitivity of breast cancer MCF-7/adr cells to adriamycin
Wenxian CUI ; Keqing XU ; Yuanguo LI ; Guijin CHEN ; Xiaoqun LU
China Oncology 2016;26(11):908-915
Background and purpose: AMP-activated protein kinase (AMPK) plays an important role in the regulation of cell metabolism and energy balance and is associated with cell proliferation, survival and multiple signaling pathways. Recent reports found that AMPK is involved in tumor suppression and drug resistance. The aim of this study was to explore the effect of AMPK on the anti-tumor effect of adriamycin and underlying mechanism in breast cancer MCF-7/adr cells. Methods:The anti-proliferative effects of adriamycin was detected by methyl thiazolyl tetrazolium (MTT) assay in MCF-7/adr, MCF-7/adr-vector and MCF-7/adr-AMPKαcells. The cell morphology in each group was stained with the lfuorescent dye Hoechst 33528, and the effects on apoptosis induction were examined by lfow cytometry (FCM). The intracellular concentration of adriamycin was detected by lfuorescence assay. The resis-tance-and apoptosis-related proteins were analyzed by Western blot. Results:The growth of breast cancer MCF-7/adr cells was inhibited by adriamycin in a dose-and time-dependent manner. The IC50 values at 24 and 48 h were (36.8±2.1) and (28.8±1.3) μg/mL, respectively. AMPKαover-expression enhanced the cytotoxic effect of adriamycin in MCF-7/adr-AMPKαcells in a dose-and time-dependent manner. Its IC50 values at 24 and 48 h were (16.0±0.7) and (4.2±0.2) μg/mL, respectively. Fluorescent morphological assay showed that AMPKαoverexpression contributed to adriamycin induced apoptosis in MCF-7/adr-AMPKαcells. After treatment with 1.0 μg/mL adriamycin for 48 h, the apoptosis rates of MCF-7/adr, MCF-7/adr-vector and MCF-7/adr-AMPKα cells were (12.0±1.4)%, (12.7±1.6)% and (32.0±4.2)%, respectively, indicating that overexpression of AMPKα enhanced the adriamycin-induced apoptosis in MCF-7/adr cells. Fluorescence microplate assay showed that over expression of AMPKαsigniifcantly increased the intracellular accumulation of adriamycin, in a concentration dependent manner. Western blot analysis showed that, compared with MCF-7/adr and MCF-7/adr-vector cells, the expressions of Bax, caspase-3 and cleaved PARP proteins were increased. Meanwhile, Bcl-2 and P-gp protein expressions were decreased in MCF-7/adr-AMPKαcells. Furthermore, the release of cytochrome c from mitochondria into the cytosol was also observed in MCF-7/adr-AMPKαcells. Conclusion:AMP-Kαoverexpression can enhance the chemosensitivity of breast cancer MCF-7/adr cells to adriamycin through inhibiting the drug effux transporter and regulating the expression of apoptosis-related proteins.
4.Tamsulosin versus nifedipine for the management of lower ureteral stones: a meta-analysis
Chong LI ; Juan WANG ; Wenjuan CAO ; Yanjun GAO ; Keqing LU ; Zhiping WANG
Chinese Journal of Urology 2015;36(11):847-853
Objective To compare the efficacy and safety of tamsulosin with nifedipine for medical expulsive therapy (MET) in patients with lower ureteral stones (LUS).Methods Randomized controlled trials(RCTs) in comparison of tamsulosin and nifedipine in treatment of LUS published in Pubmed, Cochrane Library,Embase,CNKI,CBM, Wanfang and VIP from databases establishment to July 2015 were retrieved.According to Cochrane handbook, the quality of included RCTs were assessed, and the relevant data including the number of participants, stone size, stone expulsion rate, time to stone expulsion, drug-related side effect,the incidence of ESWL or ureteroscopy lithotripsy (URSL) after MET and analgesic dose were extracted by two reviewers independently.The statistical software RevMan 5.2 was used for meta-analysis with regard to the stone expulsion rate, the incidence of ESWL or URSL and adverse effects.This study lasted more than one month from June to July 2015.Results A total of 13 RCTs with 4 831 patients were eligible.The results showed that the stone expulsion rate and the incidence rate of ESWL or URSL after MET were 92% (2 221/2 423) and 8% (27/333) in the tamsulosin group,and 73% (1 748/2 408) and 20% (67/328) in the nifedipine group.There are statistically significant differences (RR =1.24,95 % CI 1.13-1.37, P < 0.05;RR =0.40,95 % CI 0.27-0.60, P < 0.05, respectively).The subgroup analysis indicated no statistically significant differences in drug-related adverse effects between tamsulosin and nifedipine with 5% (99/1 804)and 7% (117/1 796) minor adverse effects respectively and less than 1% severe adverse effects in both groups (RR =0.85,95% CI 0.65-1.10, P =0.21;RR =0.49,95 % CI 0.09-2.59, P =0.40).Conclusion Compared to nifedipine, tamsulosin has higher stone expulsion rate and lower incidence rates for ESWL or URSL.Since there was no obvious adverse effects, tamsulosin could be considered as a preferable option for patients with LUS.
5.Phosphorylated TDP-43 Staging of Primary Age-Related Tauopathy.
Xiaoling ZHANG ; Bing SUN ; Xing WANG ; Hui LU ; Fangjie SHAO ; Annemieke J M ROZEMULLER ; Huazheng LIANG ; Chong LIU ; Jiadong CHEN ; Manli HUANG ; Keqing ZHU
Neuroscience Bulletin 2019;35(2):183-192
Primary age-related tauopathy (PART) is characterized by tau neurofibrillary tangles (NFTs) in the absence of amyloid plaque pathology. In the present study, we analyzed the distribution patterns of phosphorylated 43-kDa TAR DNA-binding protein (pTDP-43) in the brains of patients with PART. Immunohistochemistry and immunofluorescence double-labeling in multiple brain regions was performed on brain tissues from PART, Alzheimer's disease (AD), and aging control cases. We examined the regional distribution patterns of pTDP-43 intraneuronal inclusions in PART with Braak NFT stages > 0 and ≤ IV, and a Thal phase of 0 (no beta-amyloid present). We found four stages which indicated potentially sequential dissemination of pTDP-43 in PART. Stage I was characterized by the presence of pTDP-43 lesions in the amygdala, stage II by such lesions in the hippocampus, stage III by spread of pTDP-43 to the neocortex, and stage IV by pTDP-43 lesions in the putamen, pallidum, and insular cortex. In general, the distribution pattern of pTDP-43 pathology in PART cases was similar to the early TDP-43 stages reported in AD, but tended to be more restricted to the limbic system. However, there were some differences in the distribution patterns of pTDP-43 between PART and AD, especially in the dentate gyrus of the hippocampus. Positive correlations were found in PART between the Braak NFT stage and the pTDP-43 stage and density.
Aged
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Aged, 80 and over
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Aging
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metabolism
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pathology
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Brain
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metabolism
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pathology
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DNA-Binding Proteins
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metabolism
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Disease Progression
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Female
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Humans
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Immunohistochemistry
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Inclusion Bodies
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pathology
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Male
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Middle Aged
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Neurofibrillary Tangles
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metabolism
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pathology
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Neurons
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metabolism
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pathology
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Severity of Illness Index
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Tauopathies
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metabolism
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pathology
6.Interaction Between Variations in Dopamine D2 and Serotonin 2A Receptor is Associated with Short-Term Response to Antipsychotics in Schizophrenia.
Liansheng ZHAO ; Huijuan WANG ; Yamin ZHANG ; Jinxue WEI ; Peiyan NI ; Hongyan REN ; Gang LI ; Qiang WANG ; Gavin P REYNOLDS ; Weihua YUE ; Wei DENG ; Hao YAN ; Liwen TAN ; Qi CHEN ; Guigang YANG ; Tianlan LU ; Lifang WANG ; Fuquan ZHANG ; Jianli YANG ; Keqing LI ; Luxian LV ; Qingrong TAN ; Yinfei LI ; Hua YU ; Hongyan ZHANG ; Xin MA ; Fude YANG ; Lingjiang LI ; Chuanyue WANG ; Huiyao WANG ; Xiaojing LI ; Wanjun GUO ; Xun HU ; Yang TIAN ; Xiaohong MA ; Jeremy COID ; Dai ZHANG ; Chao CHEN ; Tao LI ; Chinese Antipsychotics Pharmacogenomics Consortium
Neuroscience Bulletin 2019;35(6):1102-1105