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Zhao S.M.,Peking Union Medical College | Wang J.X.,Peking Union Medical College | Liu H.,Beijing Hospital | Peng M.T.,National Center for Clinical Laboratory
Zhongguo shi yan xue ye xue za zhi / Zhongguo bing li sheng li xue hui = Journal of experimental hematology / Chinese Association of Pathophysiology | Year: 2013

The aim of this study was to develop an ex vivo cell culture system for establishing the hematological malignancy model. Mouse bone marrow cells were transfected with GFP-expressed retroviral vectors encoding various leukemia/lymphoma-associated fusion proteins (TEL-PDGFR, Rabaptin5-PDGFR, p210BCR-ABL, AML1-ETO, NPM-ALK). After transfection, the cells were cultured in IMDM containing 10% FCS without growth factors, or with one of the following growth factor combinations: (1) murine c-kit ligand (KL) plus human flt3 ligand (FL); (2) IL-3, thrombopoietin, G-CSF, and hyper-IL-6 (3/T/G/H6); (3) KL/FL plus 3/T/G/H6. The flow cytometry was used to detect the ability of combinations of growth factors to complement the oncogene fusion protein to support self-renewal of the transfected cells. The results showed that the transfected cells could be amplified sustainably in the logarithmic growth way. The indicated combination of c-Kit ligand (KL) with flt-3 ligand (FL) supported the self-renewal of the marrow cells transfected with vectors encoding TEL-PDGFR, Rabaptin5-PDGFR, AML1-ETO and NPM-ALK, in addition to KL/FL, the self-renewal of p210 BCR-ABL transfected-marrow cells also required IL-3. The morphology of cells emerged from culture can be the predictor of the corresponding oncogene-associated malignancy. It is concluded that this study establishes a culture system ex vivo which provides a generalized method for studying hematological malignancies, and may facilitate the screening for therapeutic agents.

Dai D.-P.,Key Laboratory of Geriatrics | Zhou X.-Y.,Key Laboratory of Geriatrics | Xiao Y.,Key Laboratory of Geriatrics | Xu F.,Beijing Hospital | And 12 more authors.
European Journal of Clinical Investigation | Year: 2010

Background A mutation in MEF2A (myocyte enhancer factor-2A) had been reported to be the first gene linked directly to coronary artery disease (CAD). However, an opposing opinion was proposed recently that MEF2A mutations are not a common cause of sporadic CAD. In this study, we screened exon 11 of the MEF2A gene in people of the Han nationality in China and finished some functional analysis of found variations. Materials and methods A gene structural investigation of MEF2A in 257 CAD patients and 154 control individuals were developed in this study. Subsequently, typical MEF2A variations were cloned and expressed in HeLa or 293T cell line to illustrate whether found structure changes could influence the main biological functions of these proteins. At last, another set of gene structural screen was initialized to get more reliable conclusions. Results Totally 16 different variations were detected in exon 11 of this gene in the first set of gene structural screen. By cloning and expressing typical MEF2A proteins in cultured cells, all the acquired MEF2A variations had transcriptional activation capabilities and subcellular localization patterns similar to those of the wild-type protein. Further larger scale genetic screening also revealed that the reported genetic variations of MEF2A did not differ significantly between CAD patients and healthy controls. Conclusions Our results reveal that structural changes of exon 11 in MEF2A are not involved in sporadic CAD in the Han population of China. © 2010 Stichting European Society for Clinical Investigation Journal Foundation.

Ceriotti F.,Istituto Scientifico Universitario ffaele | Queralto J.,Servei de Bioquimica | Ziyu S.,National Center for Clinical Laboratory | Ozarda Y.,Uludag University | And 3 more authors.
Clinical Chemistry and Laboratory Medicine | Year: 2010

Background: Aspartate aminotransferase (AST), alanine aminotransferase (ALT) and γ-glutamyl transferase (GGT) measurements are important for the assessment of liver damage. The aim of this study was to define the reference intervals (RIs) for these enzymes in adults, paying attention to standardization of the methods used and careful selection of the reference population. Methods: AST, ALT and GGT were measured with commercial analytical systems standardized to the IFCC-recommended reference measurement systems. Three centers (two in Italy and one in China) measured their own freshly collected samples; one of these centers also measured frozen samples from the Nordic Countries RI Project and from a Turkish center. RIs were generated using non-parametric techniques from the results of 765 individuals (411 females and 354 males, 18-85 years old) selected on the basis of the results of other laboratory tests and a specific questionnaire. Results: AST results from the four regions (Milan, Beijing, Bursa and Nordic Countries) were statistically different, but these differences were too small to be clinically relevant. Likewise, differences between the upper reference limits for genders was only 1.7 U/L (0.03 μkat/L), allowing a single RI of 11-34 U/L (0.18-0.57 μkat/L) to be defined. Interregional differences were not statistically significant for ALT, but partitioning was required due to significant gender differences. RIs for ALT were 8-41 U/L (0.13-0.68 μkat/L) for females and 9-59 U/L (0.15-0.99 μkat/L) for males, respectively. The upper reference limits for GGT from the Nordic Country population were higher than those from the other three regions and results from this group were excluded from final calculations. The GGT RIs were 6-40 U/L (0.11-0.66 μkat/L) for females and 12-68 U/L (0.20-1.13 μkat/L) for males, respectively. Conclusions: For AST and ALT, the implementation of common RIs appears to be possible, because no differences between regions were observed. However, a common RI for GGT that is applicable worldwide appears unlikely due to differences among populations. © 2010 by Walter de Gruyter.

Zhang L.-Q.,Peking Union Medical College | Zhang L.-Q.,National Center for Clinical Laboratory | Dai D.-P.,Beijing Institute of Geriatrics | Gan W.,Peking Union Medical College | And 5 more authors.
Molecular and Cellular Biochemistry | Year: 2012

The molecule 8-oxo-7,8-dihydroguanine (8-oxoGua), an oxidized form of guanine, can pair with adenine or cytosine during nucleic acid synthesis. Moreover, RNA containing 8-oxoGua causes translational errors, thus leading to the production of abnormal proteins. Human NUDT5, a MutT-related protein, catalyzes the hydrolysis of 8-oxoGDP to 8-oxoGMP, thereby preventing misincorporation of 8-oxoGua into RNA. To investigate the biological roles of NUDT5 in mammalian cells, we established cell lines with decreased level of NUDT5 expression. In NUDT5 inhibited cells, the RNA oxidation was not significantly higher than that of normal cells. However, the cell cycle G1 phase was significantly delayed, and cell numbers in both S and G2/M phases were reduced, indicating that cell proliferation was hampered by NUDT5 suppression. Key proteins for preventing the G1-S transition, including p53, p16, and Rb were increased, while the Rb phosphorylation was decreased. These results suggested that the NUDT5 protein may play significant roles in regulating the G1-S transition in mammalian cells. © 2011 Springer Science+Business Media, LLC.

Bao X.,Laboratory of An Hui Chest Hospital | Wang Q.,Laboratory of An Hui Chest Hospital | Chen G.,Laboratory of An Hui Chest Hospital | Wang Q.,Anhui Medical University | And 2 more authors.
Angiology | Year: 2014

We evaluated the association between serum uric acid (SUA) levels and prehypertension in a Chinese population. A cross-sectional study was performed during 2008 to 2010, and a total of 11199 participants without hypertension or other cardiovascular diseases (CADs), aged ≥35 years, were available for analysis. After adjusting for age, alcohol consumption, smoking status, body mass index, diabetes, total cholesterol, triglycerides, serum creatinine, the odds ratios (ORs) and 95% confidence intervals (CIs) of the prehypertension from the lowest (referent) to the highest levels of SUA were 1.00 (95% CI, 0.91-1.10), 1.12 (95% CI, 1.03-1.21), 1.17 (95% CI, 1.09-1.27), and 1.25 (95% CI, 1.13-1.39; linear trend P =.002). This association persisted in subgroup analysis by gender and was also consistent with separate analysis by classification of age, smoking status, alcohol usage, overweight, and diabetes mellitus. Independent of other cardiovascular risk factors, higher SUA levels are positively associated with prehypertension in a Chinese population without hypertension and CADs. Prospective trials should evaluate interventions that lower the SUA levels. © The Author(s) 2013.

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