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.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
8.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
9.The clinical characteristics and therapy of multiple endocrine neoplasia type 2.
Yi XIE ; Han-zhong LI ; Shi RONG ; Yu ZHU
Chinese Journal of Surgery 2004;42(18):1096-1099
OBJECTIVETo study the clinical characteristics and treatment of multiple endocrine neoplasia (MEN) type 2.
METHODSThe clinical features, diagnosis and treatment of from 1980 to 2002 8 cases of multiple endocrine neoplasia type 2 from Peking Union Medical College Hospital were reviewed and analyzed in clinical features, diagnosis and treatment retrospectively.
RESULTSSeven cases were with paroxysmal hypertension, the highest blood pressure was over 200 mm Hg (1 mm Hg = 0.133 kPa). Tumor was found in 3 cases. The diagnosis was confirmed by B-ultrasonography, CT and urine catecholamine test: six cases with MEN 2a 2 with MEN 2b. Six cases were medullary carcinoma of thyroid with or without parathyroidoma or hyperplasia, 1 with multiple mucosal neuromata. One case was pheochromocytoma with marfan's syndrome; 7 cases were with bilateral adrenal tumor. Total resection or resection ectomy was performed on different stages on adrenal gland, parathyroid tubercle. Average follow-up was 9 years. And the feedback was good.
CONCLUSIONThe diagnosis of multiple endocrine neoplasia type 2 depends on endocrine biochemical tests, B-ultrasonography and CT. When the pheochromocytoma and the other tumor exists at the same time, operation is the primary treatment, and it is better to remove the pheochromocytoma at the first.
Adolescent ; Adrenal Gland Neoplasms ; diagnosis ; surgery ; Adrenalectomy ; Adult ; Child ; Female ; Humans ; Male ; Middle Aged ; Multiple Endocrine Neoplasia Type 2a ; diagnosis ; surgery ; Parathyroid Neoplasms ; diagnosis ; surgery ; Parathyroidectomy ; Pheochromocytoma ; diagnosis ; surgery ; Retrospective Studies ; Thyroid Neoplasms ; diagnosis ; surgery ; Thyroidectomy
10.Solitary pulmonary nodules: comparison of multi-slice computed tomography perfusion study with vascular endothelial growth factor and microvessel density.
Rong-jie BAI ; Xiao-guang CHENG ; Hui QU ; Bao-zhong SHEN ; Ming-jun HAN ; Zhen-hua WU
Chinese Medical Journal 2009;122(5):541-547
BACKGROUNDThe solitary pulmonary nodule (SPN) is one of the most common findings on chest radiographs. The objectives of clinical practice are to differentiate malignant nodules from benign nodules in the least invasive way and to make a specific diagnosis. This study was aimed to evaluate the correlation between perfusion imaging features and microvessel density (MVD) and vascular endothelial growth factors (VEGF) in SPNs using multi-slice computed tomography (MSCT); and to provide the theoretical basis for SPN blood flow pattern and blood flow quantitative features. Also, the study called for the discussion of the method's clinical application value in the differential diagnosis of benign and malignant SPNs.
METHODSSixty-eight patients with SPN underwent multi-location dynamic contrast enhanced (nonionic contrast material was administrated via the antecubital vein at a rate of 4 ml/s) MSCT. Precontrast and postcontrast attenuations on every scan was studied. Perfusion, peak height, and the ratio of the peak height of the SPN to that of the aorta were analyzed. Perfusion was calculated using the maximum gradient of the time-density curves (TDC) and the peak height of the aorta. The quantitative parameters (perfusion, peak height, ratio of peak height of the SPN to that of the aorta) of the blood flow pattern were compared with MVD and the VEGF expression of immunohistochemistry.
RESULTSThe perfusion peak heights of malignant ((96.15 +/- 11.55) HU) and inflammatory ((101.15 +/- 8.41) HU) SPNs were significantly higher than those of benign ((47.24 +/- 9.15) HU) SPNs (P < 0.05, P < 0.05). Ratios of SPN-to-aorta of malignant and inflammatory SPNs were significantly higher than those of benign SPNs (P < 0.05, P < 0.05). No significant differences were found between the peak height and SPN-to-aorta ratio of malignant SPNs and inflammatory SPNs (P > 0.05, P > 0.05). The precontrast densities of inflammatory SPNs were lower than those of malignant SPNs (P < 0.05). Perfusion values of malignant and inflammatory SPNs were significantly higher than those of the benign SPNs (P < 0.05, P < 0.05). The VEGF positive expressions appeared in 32 patients with malignant SPNs and 2 patients with benign SPNs, and the average value of the MVD was higher in patients with malignant SPNs (36.88 +/- 6.76) than in patients with either benign (4.51 +/- 0.60) or inflammatory (26.11 +/- 5.43) SPNs (P < 0.05, P < 0.05). There were statistically significant correlations between the CT perfusion feature and the MVD. The highest correlation was between the peak height of SPN and the MVD (r = 0.657, P < 0.05).
CONCLUSIONSTumor microvessel density and VEGF expression facilitate the exploration of the pathophysiological basis of CT perfusion in SPNs. Multi-slice CT perfusion has shown strong positive correlations with angiogenesis in SPNs.
Adult ; Aged ; Female ; Humans ; Immunohistochemistry ; In Vitro Techniques ; Lung ; diagnostic imaging ; metabolism ; pathology ; Male ; Microvessels ; pathology ; Middle Aged ; Neovascularization, Pathologic ; Perfusion Imaging ; Solitary Pulmonary Nodule ; diagnostic imaging ; metabolism ; pathology ; Tomography, X-Ray Computed ; methods ; Vascular Endothelial Growth Factor A ; analysis