2.The study of hematopoietic cell reaction to interleukin-15 in children with myelodysplastic syndrome
han-rong, CHENG ; ming-zhen, CHEN ; ri-ling, CHEN ; de-yuan, ZHENG ; zhong-lv, YE
Journal of Applied Clinical Pediatrics 1986;0(01):-
Objective To investigate children′s myelodysplastic hematopoietic cells reaction to interleukin (IL)-15.Methods CD 34 + cells in bone marrow from 18 myelodysplast syndrome(MDS) patients were purified by an immunomagnetic beads sorting system. Apoptosis of hematopoietic precursors was assayed by propidium iodine staining and flow cytometric analysis.Results On 8th cultured day,when IL-15 concentration was between 0-100 ng/ml,it could suppress apoptosis of hematopoietic cells in MDS patients in a dose-and- time dependent manner. IL-15 in study group significanthy lower than that of control group.Conclusion IL-15 may partly suppress apoptosis of hematopoietic cells in MDS patients.
3.Research on mechanism of chloroquine phosphate-induced U937 cell apoptosis
Jia, LIU ; Fang-yuan, CHEN ; Hai-rong, WANG ; Ji-hua, ZHONG ; Li-ming, WANG ; Hua, ZHONG ; Jie-ying, HAN ; Ren-rong OUYANG
Journal of Shanghai Jiaotong University(Medical Science) 2009;29(7):813-816
Objective To observe the effects of chloroquine phosphate on apoptosis of leukemic cell line U937, and investigate whether chloroquine phosphate induces leukemic cell apoptosis by normalizing protein PNAS-2's abnormal subcellular location. Methods Chloroquine phosphate of different concentrations were added into culture fluid of leukemic cell line U937 at logarithmic phase. MTr was used to measure cell proliferation, flow cytometry and laser confocal microscopy were applied to detect cell apoptosis, and immunofluorescence technology was employed to observe the effects of chloroquine phosphate on the changes of subcellular location of protein PNAS-2. Results Apoptosis of leukemic cell line U937 was significantly induced by 50 μg/mL chloroquine phosphate, and subcellular location of protein PNAS-2 was changed. Conclusion Chlorequine phosphate can induce apoptosis of leukemic cell line U937, and the mechanism may be related to the normalization of PNAS-2's abnormal subcellular location in U937 cell line. Chloroquine phosphate has the potential to be used in leukemic therapy.
5.Optimization of processing technology for xanthii fructus by UPLC fingerprint technique and contents of toxicity ingredient.
Yan-Quan HAN ; Yan HONG ; Lun-Zhu XIA ; Jia-Rong GAO ; Yong-Zhong WANG ; Yan-Hua SUN ; Jin-Hai YI
China Journal of Chinese Materia Medica 2014;39(7):1248-1254
The experiment's aim was to optimize the processing technology of Xanthii Fructus which through comparing the difference of UPLC fingerprint and contents of toxicity ingredient in water extract of 16 batches of processed sample. The determination condition of UPLC chromatographic and contents of toxicity ingredient were as follows. UPLC chromatographic: ACQUITY BEH C18 column (2.1 mm x 100 mm, 1.7 microm) eluted with the mobile phases of acetonitrile and 0.1% phosphoric acidwater in gradient mode, the flow rate was 0.25 mL x min(-1) and the detection wavelength was set at 327 nm. Contents of toxicity ingredient: Agilent TC-C18 column (4.6 mm x 250 mm, 5 microm), mobile phase was methanol-0.01 mol x L(-1) sodium dihydrogen phosphate (35: 65), flow rate was 1.0 mL x min(-1), and detection wavelength was 203 nm. The chromatographic fingerprints 16 batches of samples were analyzed in using the similarity evaluation system of chromatographic, fingerprint of traditional Chinese medicine, SPSS16.0 and SIMCA13.0 software, respectively. The similarity degrees of the 16 batches samples were more than 0.97, all the samples were classified into four categories, and the PCA showed that the peak area of chlorogenic acid, 3,5-dicaffeoylquinic acid and caffeic acid were significantly effect index in fingerprint of processed Xanthii Fructus sample. The outcome of determination showed that the toxicity ingredient contents of all samples reduced significantly after processing. This method can be used in optimizing the processing technology of Xanthii Fructus.
Caffeic Acids
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analysis
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toxicity
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Chemistry, Pharmaceutical
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Chromatography, High Pressure Liquid
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methods
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Drugs, Chinese Herbal
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analysis
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toxicity
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Quinic Acid
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analogs & derivatives
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analysis
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toxicity
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Xanthium
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chemistry
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classification
6.Effect of IL-15 on the proliferation, differentiation and anti-apoptosis of CD34+ cells in patients with MDS.
Ming-Zheng CHENG ; Zhong-Lu YE ; Kang-Rong CAI ; Xiu-Lan HUANG ; Ri-Ling CHENG ; Han-Rong CHEN
Journal of Experimental Hematology 2005;13(4):620-623
To study the effect of interleukin-15 (IL-15) on the proliferation, differentiation and apoptosis of MDS CD34(+) cells, CD34(+) cells of high enrichment were separated by MACS system, and cultured in liquid media with different concentration of IL-15 in treated group and without IL-15 in the control group. Apoptosis of hematopoietic precursors was assayed by propidium iodine staining and cell by FCM, and the other MDS CD34(+) cells were stained by cytochemical staining after culture. The results showed that after culture with IL-15 the proliferation and differentiation of MDS CD34(+) cells were obviously promoted. It was found the every lineage of mature cells developed, the expressions of cell surface antigens CD71, CD33 and CD19 all increased in the MDS CD34(+) cell treated with IL-15. It is suggested that IL-15 stimulates the proliferation and differentiation of MDS CD34(+) cells, and partly shows anti-apoptosis effects which may be applicable to the therapy MDS.
Antigens, CD
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immunology
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Antigens, CD19
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immunology
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Antigens, CD34
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immunology
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Antigens, Differentiation, Myelomonocytic
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immunology
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Apoptosis
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drug effects
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Bone Marrow Cells
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drug effects
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immunology
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pathology
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Cell Cycle
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drug effects
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Cell Differentiation
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drug effects
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Cell Proliferation
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drug effects
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Cells, Cultured
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Flow Cytometry
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Humans
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Interleukin-15
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pharmacology
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Microscopy, Fluorescence
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Myelodysplastic Syndromes
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blood
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immunology
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pathology
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Receptors, Transferrin
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immunology
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Sialic Acid Binding Ig-like Lectin 3
7.Expression spectra of apoptosis-related gene pnas-2.
Hai-Rong WANG ; Jian-Yi ZHU ; Chun-Hong GU ; Hua ZHONG ; Ji-Hua ZHONG ; Jie-Ying HAN ; Fang-Yuan CHEN ; Ren-Rong OUYAN
Journal of Experimental Hematology 2008;16(2):282-285
To explore the expression spectra of apoptosis-related gene pnas-2 in normal tissues and acute leukemia (AL) patient tissues, the expressions of pnas-2 gene in tissues including heart, brain, placenta, lung, liver, skeletal muscle, kidney, pancreas, spleen, lymph node, thymus, leukocyte, bone marrow and fetal liver were detected by Northern blot. The expressions of pnas-2 in samples including 44 de novo, 9 non-CR, 27 CR and 12 relapsed AL patients were measured by real-time RT-PCR and Northern blot, and the expression levels of pnas-2 in normal and tumor tissues from 31 patients with malignancies were also detected. The results showed that pnas-2 was not expressed in the most tissues except in placenta. The results of real-time PCR indicated that pnas-2 expressions in samples of de novo, non-CR and relapsed patients ware significantly higher than that in CR, tumor tissues and normal tissues. In serial monitoring of 7 AL patients, the expression level of pnas-2 was high at first visit examination, but remarkably decreased after remission, and the pnas-2 expression level increased again when relapsed. It is concluded that the pnas-2 is specifically up-regulated in acute leukemia patients, which might be an oncogene and participate in leukemogenesis.
Acute Disease
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Apoptosis
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genetics
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Apoptosis Regulatory Proteins
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genetics
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metabolism
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Biomarkers, Tumor
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genetics
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Gene Expression Regulation, Leukemic
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Humans
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Leukemia
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pathology
8.Preparation and identification of monoclonal antibody against PNAS-2 protein.
Jia LIU ; Fang-Yuan CHEN ; Hai-Rong WANG ; Ji-Hua ZHONG ; Hua ZHONG ; Jie-Ying HAN ; Ren-Rong OUYANG
Journal of Experimental Hematology 2009;17(5):1269-1272
This study was purposed to prepare and primarily identify the specific monoclonal antibodies (McAbs) against the apoptosis related protein PNAS-2 so as to provide the essential tool for study of PNAS-2 function. The McAbs against PNAS-2 were prepared via the immunization of mice, cell fusion and cloning using synthetic peptide of PNAS-2 as immunogen; the specificity, titer and subtype of McAb were detected by Western blot, ELISA and immunofluorescence. The results showed that the stable hybridoma cell line S-31-7 producing McAbs against PNAS-2 protein was successfully obtained. The immunoglobulin of the McAb was identified to be IGg1lambda. The titer of ascetic fluid fled McAb were 1:8,000. A single specific band with 28 kD was shown in Western blot test, and the antigen recognized was present in cell cytoplasm by immunofluorescence. In conclusion, the obtained McAb against PNAS-2 displays strong specificity and high titer, which may be applied to the advanced research on PNAS-2 protein.
Animals
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Antibodies, Monoclonal
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immunology
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isolation & purification
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Antibody Specificity
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immunology
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Apoptosis Regulatory Proteins
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immunology
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Female
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Mice
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Mice, Inbred BALB C
9.Effects of Red Orpiment on Cell Morphology and Expression of PML mRNA and Protein in NB4, and HL-60 Cells
Lu ZHONG ; Fang-Yuan CHEN ; Jie-Ying HAN ; Nian-Xian SHAO ; Ren-Rong OUYANG
Journal of Experimental Hematology 2001;9(3):223-227
In this study the effects of red orpiment on NB4 and HL-60 cells were tried to determine. Semi-quantitative RT-PCR to determine the PML mRNA expression, immuno-fluorcscence study, together with the fluorescence stain and morphological observations were used. The results showed that: (1) red orpiment induces apoptosis morphologically in NB4 and HL-60 cells, the morphology of typical apoptosis can be seen in NB4 and HL-60 cells after 12 hours of treatment with red orpiment. Through the Wright's stain, we can see the extensive membrane blebbing, condensation of heterochromatin, and cell fragmentation, apoptotic body appearing. Many dead cells can be found on the second day. (2) in NB4 cells, red orpiment is shown to induce the PML-RARalpha chimera disappearance and to reorganize then to degradation of PML nuclear bodies which also seen in HL-60 cells, indirect immunofluorescence staining of PML with a specific monoclonal antibody was performed in control and treated cells. In NB4 cells, the control was diffusely microspeckled pattern of immunoreactivity. Upon red orpiment treatment, the microspeckled pattern disappeared, PML protein reversed into normal location. and the the size and the brightness of the particles were increased obviously. The normal nuclear distribution of PML protein was seen in untreated HL-60 cells. After treatment with red orpiment, in the nuclei of HL-60 cells, the size and the brightness of the particles were also increased. After two days of treatment with red orpiment, the immunofluorescent particles in cells almost disappeared. (3) the expression of PML mRNA is not changed in red orpiment-treated cells, RT-PCR to determine the PML mRNA expression in NB4 and HL-60 cells treated with red orpiment, the expression results are similar to the controls, that to say, the PML mRNA lever is unaffected. It was concluded that, red orpiment induced PML to play the effects of induce apoptosis in leukemia cells at the translational level and inhibited the proliferation of leukemia cells.
10.A cross-sectional study on injuries in residents at the community level of Zhejiang.
Min YU ; Li-ming CONG ; Lai-rong XU ; Zi-cheng XIA ; Cheng-xing HAN ; Yan MA ; Jie-ming ZHONG
Chinese Journal of Epidemiology 2003;24(8):681-683
OBJECTIVETo study the incidence of injuries among residents living in the communities in Zhejiang.
METHODSA household questionnaire survey was conducted to 16 899 residents in four communities.
RESULTSThe incidence of injuries was 16.11% in total, higher in urban area than in rural area and higher in men than in women. The leading causes of injuries were falls (4.80%), knife-cutting or by sharp articles (3.86%), traffic accident (2.07%), collision (1.99%), scalds (1.13%) and animal bites (1.06%). Peak incidence of age group was different in different injuries, such as traffic accident was high in 45 - 60 age group and falls was high in 5 and 60 age group. Direct economic loss for each case with injury would cost 324.21 RMB yuan.
CONCLUSIONInjury was common and frequently occurred among residents at the community level which seriously on danger their health and families.
Accidental Falls ; statistics & numerical data ; Accidents, Traffic ; statistics & numerical data ; Adolescent ; Adult ; Aged ; Animals ; Bites and Stings ; epidemiology ; Burns ; epidemiology ; Child ; China ; epidemiology ; Community Health Services ; Dogs ; Female ; Humans ; Male ; Middle Aged ; Prevalence ; Wounds and Injuries ; epidemiology ; mortality