1.Involvement of the mitochondrion-dependent and the endoplasmic reticulum stress-signaling pathways in isoliquiritigenin-induced apoptosis of HeLa cell.
Xuan YUAN ; Bo ZHANG ; Lu GAN ; Zhen Hua WANG ; Ba Cui YU ; Liang Liang LIU ; Qiu Sheng ZHENG ; Zhi Ping WANG
Biomedical and Environmental Sciences 2013;26(4):268-276
OBJECTIVEIsoliquiritigenin (ISL), a licorice chalconoid, is considered to be a bioactive agent with chemopreventive potential. This study investigates the mechanisms involved in ISL-induced apoptosis in human cervical carcinoma HeLa cells.
METHODSCell viability was evaluated using a 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl-tetrazolium bromide (MTT) assay. Apoptosis was determined by flow cytometry using an Annexin V-FITC Apoptosis Detection Kit. The intracellular ROS levels were assessed using a 2, 7-dichlorofluorescein probe assay. The mitochondrial membrane potential was measured with the dual-emission potential-sensitive probe 5, 5', 6, 6'-tetra-chloro-1, 1', 3, 3'-tetraethyl-imidacarbocyanine iodide (JC-1). The degradation of poly-ADP-ribose polymerase (PARP) protein, the phosphorylation of PKR-like ER kinase (PERK), the phosphorylation of the α-subunit of eukaryotic initiation factor 2 (eIF2α), the expression of the 78 kD glucose-regulated protein (GRP 78), and the activation of caspase-12 were analyzed via western blot analysis.
RESULTSISL significantly inhibited the proliferation, the increase in ROS levels and apoptotic rates of HeLa cells in a concentration-dependent manner. Moreover, ISL induced mitochondrial dysfunction, caspase activation, and PARP cleavage, which displayed features of mitochondria dependent on apoptotic signals. Besides, exposure of HeLa cells to ISL triggered endoplasmic reticulum (ER) stress, as indicated by the increase in p-eIF2α and GRP78 expression, ER stress-dependent apoptosis is caused by the activation of ER-specific caspase-12.
CONCLUSIONThe findings from our study suggest that ISL-induced oxidative stress causes HeLa cell apoptosis via the mitochondrion-dependent and the ER stress-triggered signaling pathways.
Aldehyde Reductase ; antagonists & inhibitors ; Apoptosis ; drug effects ; Cell Survival ; drug effects ; Chalcones ; pharmacology ; therapeutic use ; Chemoprevention ; Drug Screening Assays, Antitumor ; Endoplasmic Reticulum Stress ; drug effects ; HeLa Cells ; Humans ; Mitochondria ; drug effects ; Neoplasms ; prevention & control ; Reactive Oxygen Species ; metabolism
2.Study on the epidemiological characteristics and molecular typing of Salmonella enterica subsp. enterica serovar Senftenberg in Shanghai
Xue-Bin XU ; Zhen-Gan YUAN ; Hui-Ming JIN ; Wen-Jia XIAO ; Bao-Ke GU ; Min CHEN ; Lu RAN ; Bao-Wei DIAO ; Zhi-Gang CUI ; Qing-Hua HU ; Biao KAN
Chinese Journal of Epidemiology 2009;30(9):933-937
ation were complicated, with the characteristics as the obvious decreasing number of patients, with no food-borne isolates in 2007.
3.Analysis of Cut-off Value in Screening of Thalassemia by Capillary Hemoglobin Electrophoresis for Pregnant Women from Shenzhen Region of China.
Mei HUO ; Wen-Yuan WU ; Mei LIU ; Zhi-Biao GAN ; Wei-Yu MAO ; Rong-Yao LIN ; Ai-Qin LIU ; Gui-Rong HE
Journal of Experimental Hematology 2016;24(2):536-539
OBJECTIVETo investigate the cut-off value in screening of thalassemia in pregnant women from Shenzhen region by capillary hemoglobin electrophoresis.
METHODSThe data of capillary hemoglobin electrophoresis and genetic diagnosis of thalassemia from 2122 examined prenatal women were retrospectively analyzed. Capillary hemoglobin electrophoresis and α-, β- genetic diagnosis of thalassemia were carried out for every woman. Hemoglobin electrophoresis was performed using Capillarys 2 full-automated electrophoresis instrument. Gap polymerase chain reaction and reverse dot blot were used for genetic diagnosis of thalassemia genotyping test. The cut-off value in screening of thalassemia was determined by receiver operating characteristic curve and next to analyze the value of HbA2 and HbF in screening of thalassemia using the decided cut-off value.
RESULTSThe areas under the curve (AUC(Roc)) of HbA2 for diagnosis of α-, β- thalassemia were 0.75 and 0.981 respectively, and the AUC(Roc) of HbF for diagnosis of β-thalassemia was 0.787. When HbA2 ≤ 2.55 was taken as the cut-off value of HbA2 for diagnosis of α-thalassemia, the sensitivity, specificity, positive likelihood ratio (LR(+)) and negative likelihood ratio (LR(-)) were 89.5%, 54.8%, 1.98, 0.19 respectively. When HbA2 ≥3.9 was taken as the cut off value of HbA2 for diagnosis of β-thalassemia, the sensitivity, specificity, LR(+) and LR(-) were 96.1%, 99.8% 480.5, 0.04 respectively. When HbF ≥0.75 was taken as the cut off value of HbF for diagnosis of β-thalassemia, the sensitivity, specificity, LR(+) and LR(-) were 83.6%, 61.8% respectively.
CONCLUSIONThe cut-off value in screening of thalassemia by capillarys 2 full automated electrophoresis instrument is different from that of the traditional method of hemoglobin electrophoresis, such as cellulose acetate membrane electrophoresis and agarose gel electrophoresis. Each laboratory should establish their own respective cut off value.
Area Under Curve ; China ; Electrophoresis, Capillary ; Female ; Fetal Hemoglobin ; analysis ; Genotyping Techniques ; Hematologic Tests ; Hemoglobin A2 ; analysis ; Humans ; Mass Screening ; Pregnancy ; Reference Values ; Retrospective Studies ; Sensitivity and Specificity ; alpha-Thalassemia ; diagnosis ; beta-Thalassemia ; diagnosis
4.Protective effects of shikonin on brain injury induced by carbon ion beam irradiation in mice.
Lu GAN ; ; Zhen Hua WANG ; Hong ZHANG ; ; Rong ZHOU ; ; Chao SUN ; ; Yang LIU ; ; Jing SI ; ; Yuan Yuan LIU ; ; Zhen Guo WANG
Biomedical and Environmental Sciences 2015;28(2):148-151
Radiation encephalopathy is the main complication of cranial radiotherapy. It can cause necrosis of brain tissue and cognitive dysfunction. Our previous work had proved that a natural antioxidant shikonin possessed protective effect on cerebral ischemic injury. Here we investigated the effects of shikonin on carbon ion beam induced radiation brain injury in mice. Pretreatment with shikonin significantly increased the SOD and CAT activities and the ratio of GSH/GSSG in mouse brain tissues compared with irradiated group (P<0.01), while obviously reduced the MDA and PCO contents and the ROS levels derived from of the brain mitochondria. The shikonin also noticeably improved the spatial memory deficits caused by carbon ion beam irradiation. All results demonstrated that shikonin could improve the irradiated brain injury which might resulted from its modulation effects on the oxidative stress induced by the 12C6+ ion beam.
Animals
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Antioxidants
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pharmacology
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Brain Injuries
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prevention & control
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Catalase
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metabolism
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Heavy Ion Radiotherapy
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Male
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Malondialdehyde
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metabolism
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Mice
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Naphthoquinones
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pharmacology
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Protein Carbonylation
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Radiation Injuries, Experimental
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prevention & control
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Radiation-Protective Agents
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pharmacology
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Random Allocation
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Specific Pathogen-Free Organisms
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Superoxide Dismutase
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metabolism
5.Clinical application of blood matching with hemolytic test in vitro for transfusion treatment of crisis puerpera with acute hemolytic anemia.
Min YUAN ; Cong-Hai TANG ; Wei-Wei GAN ; A-Yang WU ; Hui-Cong YANG ; Tian-Xin ZHANG ; Yan Xue HUANG ; Lu-Zhen QIU ; Hong-Pu CHEN ; Feng-Li LIN
Journal of Experimental Hematology 2014;22(4):1094-1098
This study was aimed to establish the matching method of hemolytic test in vitro, and to guide the transfusion treatment for puerpera with acute hemolytic disease. The donor's erythrocytes were sensibilized by all the antibodies in plasma of patient in vitro and were added with complement, after incubation for 6.5 hours at 38 °C, the hemolysis or no hemolysis were observed. It is safe to transfuse if the hemolysis did not occur. The results showed that when the matching difficulty happened to puerpera with acute hemolytic disease, the compatible donor could be screened by hemolytic test in vitro. There were no untoward effects after transfusion of 6 U leukocyte-depleted erythrocyte suspension. The all hemoglobin, total bilirubins, indirect bilirubin, reticulocyte, D-dimex and so on were rapidly improved in patient after transfusion , showing obvious clinical efficacy of treatment. It is concluded that when the matching results can not judge accurately compatible or incompatible through the routine method of cross matching, the agglutinated and no-hemolytic erythrocytes can be screened by hemolytic test in vitro and can be transfused with good efficacy; the hemoglobin level can be promoted rapidly, and no untoward effects occur.
Anemia, Hemolytic
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therapy
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Blood Grouping and Crossmatching
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methods
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Erythrocyte Transfusion
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methods
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Female
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Humans
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Puerperal Disorders
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therapy
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Young Adult
6.Study on HPLC fingerprint and chemical constituent difference of different species of Aurantii Fructus Immaturus.
Hong-Lian ZENG ; Zhen-Li LIU ; Zhi-Qian SONG ; Chun WANG ; Yun-Zhuo DONG ; Zhang-Chi NING ; Si-Yu ZHAO ; Yi-Song SHU ; Dan HE ; Meng-Lei WANG ; Jia-He GAN ; Yuan-Yan LIU
China Journal of Chinese Materia Medica 2016;41(17):3272-3278
This study is to establish an HPLC fingerprint by HPLC-DAD method and simultaneous quantitative analysis of 17 components of 18 batches of Citrus aurantium and 10 batches of C. sinensis. The separation was performed on an Agilent Poroshell 120 SB-C₁₈ (4.6 mm×100 mm,2.7 μm) column with the gradient elution of methanol-0.1% formic acid water, the flow was 0.6 mL•min⁻¹. The detection wavelength was set at 318 nm. The column temperature was maintained at 30 ℃. The data calculation was performed with similarity evaluation system for chromatographic fingerprint of traditional Chinese medicine (Version 2004A) together with SIMCA-P 13.0 software to clarify the differential marker between these two different species of Aurantii Fructus Immaturus. This method has good precision stability and repeatability that could provide basis for quality control and evaluation of Aurantii Fructus Immaturus.