Cell biochemistry and biophysics

Publications in this journal

• Article: Erythrocyte Membrane Properties in Patients with Essential Hypertension.

  Magdalena Kaczmarska, Maria Fornal, Franz H Messerli, Jozef Korecki, Tomasz Grodzicki, Kvetoslava Burda
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  ABSTRACT: In spite of the extensive research efforts that have been conducted over the last decades, it is still very difficult to point out genetic determinants or environmental conditions responsible for the development of essential hypertension. We searched for differences in the RBC membrane skeleton structure and O2 membrane permeability between RBCs from patients with both essential arterial hypertension and hypercholesterolemia, from patients having only hypercholesterolemia and from healthy donors. The topography of RBCs and the content of various hemoglobin forms were detected using atomic force microscopy and Mössbauer spectroscopy, respectively. We found that the membrane skeleton of RBCs from healthy donors displayed a well-known honeycomb pattern, whereas in patients with essential hypertension and/or hypercholesterolemia, who had never received anti-hypertensive therapy, it displayed a corncob pattern. Hypertensive RBCs had an oval shape and the average lateral to longitudinal diameter ratio for the changed cells (about 70 %) did not exceed 0.80. We observed that after the incubation of RBCs under high nitrogen (low O2) pressure at room temperature and then their transfer into 85 K, a content of oxyHb (deoxyHbOH) already after 1 h reached a stable level of about 85 ± 3 % (15 ± 3 %) in hypertensives, whereas in healthy individuals it showed a decrease for deoxyHbOH and an increase for oxyHb, which stabilized at a level of about 81 ± 5 % and 19 ± 5 %, respectively, only after 9 h. Quantitative analysis of the Δ(oxyHb) change estimated as the difference between the oxyHb level measured after 9 and 2 h at 85 K under low N2 pressure (to slow down oxyHb formation) was significantly higher in normotensives than in hypertensive patients with or without hypercholesterolemia, 19.9 versus -4.2, p < 0.02. Our findings indicate an impaired oxygen release by Hb in RBCs of patients with hypertension under low oxygen pressure which if present in vivo may cause hypoxemia and, in turn, further increase of blood pressure.
  Cell biochemistry and biophysics 05/2013;
• Article: DC Electric Fields Direct Breast Cancer Cell Migration, Induce EGFR Polarization, and Increase the Intracellular Level of Calcium Ions.

  Dan Wu, Xiuli Ma, Francis Lin
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  ABSTRACT: Migration of cancer cells leads to invasion of primary tumors to distant organs (i.e., metastasis). Growing number of studies have demonstrated the migration of various cancer cell types directed by applied direct current electric fields (dcEF), i.e., electrotaxis, and suggested its potential implications in metastasis. MDA-MB-231 cell, a human metastatic breast cancer cell line, has been shown to migrate toward the anode of dcEF. Further characterizations of MDA-MB-231 cell electrotaxis and investigation of its underlying signaling mechanisms will lead to a better understanding of electrically guided cancer cell migration and metastasis. Therefore, we quantitatively characterized MDA-MB-231 cell electrotaxis and a few associated signaling events. Using a microfluidic device that can create well-controlled dcEF, we showed the anode-directing migration of MDA-MB-231 cells. In addition, surface staining of epidermal growth factor receptor (EGFR) and confocal microscopy showed the dcEF-induced anodal EGFR polarization in MDA-MB-231 cells. Furthermore, we showed an increase of intracellular calcium ions in MDA-MB-231 cells upon dcEF stimulation. Altogether, our study provided quantitative measurements of electrotactic migration of MDA-MB-231 cells, and demonstrated the electric field-mediated EGFR and calcium signaling events, suggesting their involvement in breast cancer cell electrotaxis.
  Cell biochemistry and biophysics 05/2013;
• Article: Over-Expression of Small Ubiquitin-Related Modifier-1 and Sumoylated p53 in Colon Cancer.

  Hongjie Zhang, Xiaoyi Kuai, Zeyu Ji, Zhengyang Li, Ruihua Shi
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  ABSTRACT: Here we investigated whether the cellular accumulation of p53 protein caused by over-expression of small ubiquitin-related modifier-1 (SUMO-1) could be used as a predictive marker for prognosis in colon cancer. We detected SUMO-1 and p53 protein levels in 46 cases of colon cancer and adjacent tissues by immunohistochemistry and found that SUMO-1 was expressed at much higher levels in colon cancer compared with that in normal colon tissue. Immunoprecipitation and Western blot analysis revealed that the tumor suppressor p53 was present predominantly in the sumoylated rather than the non-sumoylated form in the colon cancer cell lines. A small interfering RNA targeted to SUMO-1 mRNA sequences was used to observe the levels of the p53 protein. Patients who showed high dual expressions of SUMO-1 and p53 tended to experience metastasis more frequently. These results suggest that the cellular accumulation of p53 protein caused by over-expression of SUMO-1 may be involved in tumor aggressiveness. Multivariate analysis confirmed that the high dual expression of SUMO-1 and p53 was an independent factor for evaluating prognosis. SUMO-1 may be useful as a novel target for therapy in colon cancer as well as a clinical indicator for tumor aggressiveness.
  Cell biochemistry and biophysics 05/2013;
• Article: PT-ACRAMTU, A Platinum-Acridine Anticancer Agent, Lengthens and Aggregates, but does not Stiffen or Soften DNA.

  Samrat Dutta, Matthew J Snyder, David Rosile, Kristen L Binz, Eric H Roll, Jimmy Suryadi, Ulrich Bierbach, Martin Guthold
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  ABSTRACT: We used atomic force microscopy (AFM) to study the dose-dependent change in conformational and mechanical properties of DNA treated with PT-ACRAMTU ([PtCl(en)(ACRAMTU-S)](NO3)2, (en = ethane-1,2-diamine, ACRAMTU = 1-[2-(acridin-9-ylamino)ethyl]-1,3-dimethylthiourea. PT-ACRAMTU is the parent drug of a family of non-classical platinum-based agents that show potent activity in non-small cell lung cancer in vitro and in vivo. Its acridine moiety intercalates between DNA bases, while the platinum group forms mono-adducts with DNA bases. AFM images show that PT-ACRAMTU causes some DNA looping and aggregation at drug-to-base pair ratio (r b) of 0.1 and higher. Very significant lengthening of the DNA was observed with increasing doses of PT-ACRAMTU, and reached saturation at an r b of 0.15. At r b of 0.1, lengthening was 0.6 nm per drug molecule, which is more than one fully stretched base pair stack can accommodate, indicating that ACRAMTU also disturbs the stacking of neighboring base pair stacks. Analysis of the AFM images based on the worm-like chain (WLC) model showed that PT-ACRAMTU did not change the flexibility of (non-aggregated) DNA, despite the extreme lengthening. The persistence length of untreated DNA and DNA treated with PT-ACRAMTU was in the range of 49-65 nm. Potential consequences of the perturbations caused by this agent for the recognition and processing of the DNA adducts it forms are discussed.
  Cell biochemistry and biophysics 05/2013;
• Article: Noninvasive Monitoring of Small Intestinal Oxygen in a Rat Model of Chronic Mesenteric Ischemia.

  Elaine M Fisher, Mahmood Khan, Ronald Salisbury, Periannan Kuppusamy
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  ABSTRACT: We noninvasively monitored the partial pressure of oxygen (pO2) in rat's small intestine using a model of chronic mesenteric ischemia by electron paramagnetic resonance oximetry over a 7-day period. The particulate probe lithium octa-n-butoxynaphthalocyanine (LiNc-BuO) was embedded into the oxygen permeable material polydimethyl siloxane by cast-molding and polymerization (Oxy-Chip). A one-time surgical procedure was performed to place the Oxy-Chip on the outer wall of the small intestine (SI). The superior mesenteric artery (SMA) was banded to ~30 % of blood flow for experimental rats. Noninvasive measurement of pO2 was performed at the baseline for control rats or immediate post-banding and on days 1, 3, and 7. The SI pO2 for control rats remained stable over the 7-day period. The pO2 on day-7 was 54.5 ± 0.9 mmHg (mean ± SE). SMA-banded rats were significantly different from controls with a noted reduction in pO2 post banding with a progressive decline to a final pO2 of 20.9 ± 4.5 mmHg (mean ± SE; p = 0.02). All SMA-banded rats developed adhesions around the Oxy-Chip, yet remained asymptomatic. The hypoxia marker Hypoxyprobe™ was used to validate the low tissue pO2. Brown cytoplasmic staining was consistent with hypoxia. Mild brown staining was noted predominantly on the villus tips in control animals. SMA-banded rats had an extended region of hypoxic involvement in the villus with a higher intensity of cytoplasmic staining. Deep brown stainings of the enteric nervous system neurons and connective tissue both within layers and in the mesentery were noted. SMA-banded rats with lower pO2 values had a higher intensity of staining. Thus, monitoring SI pO2 using the probe Oxy-Chip provides a valid measure of tissue oxygenation. Tracking pO2 in conditions that produce chronic mesenteric ischemia will contribute to our understanding of intestinal tissue oxygenation and how changes impact symptom evolution and the trajectory of chronic disease.
  Cell biochemistry and biophysics 05/2013;
• Article: Protein Phosphatases but not Reactive Oxygen Species Play Functional Role in Acute Amphetamine-Mediated Dopamine Release.

  Erzsebet Paszti-Gere, Judit Jakus
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  ABSTRACT: Drug abuse-induced neurodegeneration can be triggered by elevated production of reactive oxygen species (ROS). Involvement of oxidative stress in acute amphetamine (AMPH)-mediated dopamine (DA) release, however, has not been completely understood yet. In order to elucidate the dopaminergic response of PC12 cells to a single dose of 10 μM AMPH, ROS production was measured as related to the extracellular DA level. Due to the spontaneous oxidation of peroxide-sensitive fluorophore 2',7'-dichlorofluorescin diacetate (DCFH-DA) to 2',7'-dichlorofluorescein (DCF), the increase in fluorescence could not be unambiguously attributed to AMPH-triggered ROS production. Based on Amplex Red fluorescence, no ROS production was detected after acute AMPH application. Our data strongly suggest that ROS development was not the main triggering factor for immediate DA release after acute AMPH treatment. On the other hand, AMPH-induced elevation of DA levels in rat brain striatal slices was quenched by the water soluble antioxidant, N-acetylcysteine (NAC) at 10 mM. In this study, we also investigated the contribution of protein phosphatases to the AMPH-induced rat brain striatal dopaminergic response. The experimental protocol, double AMPH challenge was applied for screening the effect of NAC and cantharidin on AMPH-mediated DA release. Here we show that AMPH-mediated DA release increased nearly twofold in striatal rat brain slices pretreated for 30 min with 1000 μM cantharidin, a selective PP1 and PP2A inhibitor. These findings prove the lack of ROS inhibitory action on protein phosphatase activity in acute AMPH-mediated DA efflux.
  Cell biochemistry and biophysics 04/2013;
• Article: The Value of Expression of M2-PK and VEGF in Patients with Advanced Gastric Cancer.

  Lanning Yin, Xiang Wang, Changjiang Luo, Haipeng Liu, Ling Zhang, Hong Zhang, Youcheng Zhang
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  ABSTRACT: Glycolytic pyruvate kinase isoenzyme type M2 (M2-PK) plays a key role in tumor metabolism and energy production. Vascular endothelial growth factor (VEGF) is critical in regulating angiogenesis which is an essential process required for tumor growth and metastasis. These two genes may function in accordance with tumor development. The purpose of this study was to investigate the relationship between the expression of M2-PK and VEGF, and their association with clinicopathological features in patients with advanced gastric cancer. Expression of M2-PK and VEGF were examined in 142 cases of paraffin-embedded tissue blocks from patients with advanced gastric cancer. M2-PK expression was found to strongly correlate with that of VEGF (r = 0.718). In addition, expression of M2-PK and VEGF correlates with tumor size (p = 0.0001, and p = 0.0017, respectively), depth of invasion (p = 0.0024, and p = 0.0261, respectively), and lymph node metastasis (p = 0.036, and p = 0.028, respectively). The high expression levels of M2-PK and VEGF may indicate poor prognosis in patients with advanced gastric cancer.
  Cell biochemistry and biophysics 04/2013;
• Article: Epigenetic Signature of Early Cardiac Regulatory Genes in Native Human Adipose-Derived Stem Cells.

  Alice Pasini, Francesca Bonafè, Marco Govoni, Carlo Guarnieri, Paolo G Morselli, Hari S Sharma, Claudio M Caldarera, Claudio Muscari, Emanuele Giordano
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  ABSTRACT: Adipose-derived stem cells (ADSCs) are stromal mesenchymal stem cells isolated from lipoaspirates, and they display a broad potential to differentiate toward different lineages. The role of epigenetics in regulating the expression of their lineage-specific genes is under evaluation, however till date virtually nothing is known about the relative significance of cardiac-specific transcription factor genes in human ADSCs. The aim of this study was to investigate DNA promoter methylation and relevant histone modifications involving MEF-2C, GATA-4, and Nkx2.5 in native human ADSCs. CpG sites at the transcription start in their promoters were found unmethylated using methylation-specific PCR. Chromatin immunoprecipitation assay showed low levels of total acetylated H3 histone (acH3) and high levels of trimethylated lysine 27 in H3 histone (H3K27me3) which were associated with both GATA-4 and Nkx2.5 promoters, indicating their transcriptional repressive chromatin arrangement. On the other hand, the opposite was apparent for MEF-2C promoter. Accordingly, MEF-2C-but not GATA-4 and Nkx2.5-transcripts were evidenced in native human ADSCs. These results suggest that the chromatin arrangement of these early cardiac regulatory genes could be explored as a level of intervention to address the differentiation of human ADSCs toward the cardiac lineage.
  Cell biochemistry and biophysics 04/2013;
• Article: Construction and Characterization of a CTLA-4-Targeted scFv-Melittin Fusion Protein as a Potential Immunosuppressive Agent for Organ Transplant.

  Hailong Jin, Congran Li, Ding Li, Ming Cai, Zhouli Li, Shuang Wang, Xin Hong, Bingyi Shi
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  ABSTRACT: Immunotoxins with selective cytotoxicity are frequently used as therapeutic immunosuppressive agents in solid-organ transplantation because of their efficiency and high specificity. In this study, we present a new recombinant immunotoxin termed anti-CTLA-4-scFv-melittin prepared from Escherichia coli aimed at clearing activated T cells at the same time avoiding all-round decline in systematic immunity. This fusion protein is composed of anti-CTLA-4-scFv unit and melittin analog unit with properties of low immunogenicity and selective cytotoxicity to CTLA-4-positive T cells. In preliminary biological activity assays, our results confirmed the feasibility of activated T cell clearance strategy and there were significant differences in cell survival rates between CTLA-4-positive group and control group at all experimental concentrations of the immunotoxin. The selective cytotoxicity, low immunogenicity, and low production cost make it an attractive alternate to traditional immunosuppressants.
  Cell biochemistry and biophysics 04/2013;
• Article: Disruption of Mitochondrial Complexes in Cancer Stem Cells Through Nano-based Drug Delivery: A Promising Mitochondrial Medicine.

  C George Priya Doss, C Debajyoti, S Debottam
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  ABSTRACT: Mitochondria are the fulcrum for regulating cellular metabolism as well as apoptosis. The multi-lamellar vesicles (MLVs) liposome targeted against mitochondria can be formulated to disrupt mitochondrial integrity to attain programmed cell death of cancer stem cells (CSCs). The gold nanoparticles (GNPs) and a steroid nucleus (cyclopentanoperhydrophenanthrene ring) are encapsulated within MLV liposome that targets specifically to the CD44 receptor of the CSCs. Entering cytosol, it would bind distinctively to the malate-aspartate shuttle through a specifically designed ligand. Liposome fuses with the mito-membrane after associating with shuttle, thereby releasing both the components. The steroid disrupts mito-membrane's integrity facilitating release of cytochrome c. Thus, GNPs enter into the mitosol and interact with the mitochondrial complexes to cease cellular respiration. Since the solid nano-based pharmaceutics has shown a lot of promises as a potent anticancer therapy, the role of MLV liposome can be proved to be a better weapon to terminate malignancy.
  Cell biochemistry and biophysics 04/2013;
• Article: The Relationship Between Insulin Resistance and CpG Island Methylation of LMNA Gene in Polycystic Ovary Syndrome.

  Wang Ting, Qian Yanyan, Huang Jian, Hua Keqin, Ma Duan
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  ABSTRACT: We sought to investigate the relationship between the changes of CpG island methylation status of LMNA gene and insulin resistance in polycystic ovary syndrome (PCOS) patients. The genome-wide methylation microarray screening was done in three PCOS cases of insulin resistance and one case of a normal woman. The PCOS insulin resistance-related genes were identified as indicated by the results of gene chip screening. Then, 24 cases of insulin-resistant PCOS patients and 24 cases of normal individuals were studied to identify the effects of the candidate genes using genome-wide study of DNA from the peripheral blood analyzed by MassARRAY(®)EpiTYPER™ DNA methylation analysis technique. We found that the methylation status of CpG island in the promoter area of LMNA gene was changed. The 20 CG sites in CpG island of LMNA gene were examined using case control experiment among which 12 CpG sites differed significantly (P < 0.05) between two groups while the remaining eight CpG sites differed non-significantly. We, therefore, concluded that the changes in the hypermethylation status of CpG island of LMNA gene were related to the insulin resistance in PCOS patients, indicating that this gene may be involved in the regulation of PCOS-associated insulin resistance.
  Cell biochemistry and biophysics 04/2013;
• Article: Study of the Mechanism of Bystander Effect of KDR-CDglyTK System Mediated by Adenovirus for the Treatment of Gastric Cancer.

  Li Qiang, Li Yanping, Huang Zonghai, Chen Fei, Li Zhou, Yu Jinlong
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  ABSTRACT: We evaluated the relationship between cellular gap junction and bystander effect in gastric cancer SCG7901 cell killing by adenovirus-mediated KDR-CDglyTK system. SCG7901 and HeLa cells were transfected (MOI-100) with recombinant adenovirus carrying fusion gene. Transfection efficiency/fusion gene mRNA expression were determined; intercellular communications in the presence of apigenin were measured by fluorescence recovery after photobleaching (FRAP). Transfected/non-transfected cells were mixed at 5:95/10:90 % ratios, respectively, incubated for 24 h with/without apigenin, and for 72 h with GCV and 5-FU. Cell survival was determined by MTT method. The transfected SCG7901/HeLa cells expressed GFP. Fluorescence intensity in SCG7901 cells decreased after photobleaching but recovered over time. Fluorescence intensity in HeLa cells also decreased after photobleaching with no recovery. Following apigenin treatment, fluorescence intensity differed between SCG7901 and HeLa cells. Survival rates of SCG7901 cells differed among prodrug, apigenin, and prodrug + apigenin groups compared with controls, but no significant difference was observed for HeLa cells. We concluded that the intercellular communication in SCG7901 cells related with the gap junctions, while there was no such communication in HeLa cells. The bystander killing effect related with the intercellular gap junctions and was increased by apigenin with no such effect seen in HeLa cells.
  Cell biochemistry and biophysics 04/2013;
• Article: The Characterization of Childhood Occipital Epilepsy of Gastaut: A Study of Seven Patients.

  Xiao Mei Shu, Gui Ping Zhang, Bing Zhu Yang, Juan Li
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  ABSTRACT: Characterization of the electroclinical features and evolution of childhood occipital epilepsy of Gastaut (COE-G). Seven children were retrospectively identified as having COE-G and were followed-up clinically using EEGs. Visual manifestations were the most common ictal event. Eye-associated ictal deviation was associated with ipsilateral turning of the head and migraine-like symptoms were frequent. Hemiconvulsions occurred in two children, and only one child had secondary generalized tonic-clonic seizures. In all patients, seizures occurred while awake, while two patients also had seizures while sleeping. EEG showed five patients with occipital spike-wave discharges when their eyes were closed which disappeared once their eyes were opened. Two cases continued having frequent seizures despite antiepileptic drug treatment. These patients also displayed learning difficulties and behavioral impairments after seizure onset. COE-G is a distinctive epileptic syndrome; however, the long-term prognosis for patients with the condition is unclear.
  Cell biochemistry and biophysics 04/2013;
• Article: A Murine Model of Hepatic Veno-occlusive Disease Induced by Allogeneic Hematopoietic Stem Cell Transplantation.

  Lingyu Zeng, Licai An, Ting Fang, Bin Pan, Haiying Sun, Chong Chen, Jiang Cao, Zhenyu Li, Kailin Xu
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  ABSTRACT: Hepatic veno-occlusive disease (HVOD) is a life-threatening complication of bone marrow stem cell transplantation. The understanding of this clinical condition is hampered by the lack of suitable animal models. Here, we present a murine (BALB/c-based) model of HVOD induced by allogeneic hematopoietic stem cell transplantation (allo-HSCT). The chimerism rate of bone marrow was measured on days 5 and 10, while the chimerism rate of peripheral blood was measured on day 15 after allo-HSCT. Percentages of peripheral reticulocytes and serum levels of bilirubin and alanine aminotransferase (as liver function tests) were measured on days 5, 10, 15, 20, and 30. Livers were obtained on days 5, 10, 15, 20, and 30, and fixed in formaldehyde or glutaric dialdehyde. Liver slices were processed using the hematoxylin-eosin, Masson's trichrome, or immunohistochemistry staining, and examined by light or transmission electron microscopy. Sinusoidal damages were the earliest pathological changes occurring in the allo-HSCT-induced HVOD, followed by coagulative necrosis of liver cells. The liver cell necrosis was later attenuated and sinusoidal endothelial cell morphology improved. However, on day 30, the edema and necrosis of liver cells became aggravated again. Furthermore, sinusoidal lining cell regeneration and partly attenuated liver cell necrosis were followed by the moderate to severe central vein fibrosis. In conclusion, we have successfully established a murine model of HSCT-HVOD. This model develops moderate to severe HVOD which cannot heal without intervention.
  Cell biochemistry and biophysics 04/2013;
• Article: 2,3,4',5-Tetrahydroxystilbene-2-O-β-D-glucoside Suppresses Expression of Adhesion Molecules in Aortic Wall of Dietary Atherosclerotic Rats and Promonocytic U937 Cells.

  Yu-Qin Wang, Yan Shen, Feng Li, Chun-Hua Wang, Wei Zhang
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  ABSTRACT: We sought to investigate whether TSG suppressed the ICAM-1/VCAM-1 expression in dietary atherosclerotic rats and in Ox-LDL-induced U937 cells. For this purpose, 60 male Sprague-Dawley rats were randomly-and-equally divided into six groups. Atherosclerosis was induced by feeding rats a hyperlipidemic diet. TSG (120, 60 or 30 mg/kg/day) was administered by oral gavage. Simvastatin (2 mg/kg/day) was administered as positive control whereas physiological saline (0.9 % NaCl) served as untreated control. After 12 weeks, rats were euthanized by ethyl carbonate (1,200 mg/kg) and aortic wall samples were collected. Besides, U937 cells were stimulated for 48 h by Ox-LDL (80 μg/mL) with and without TSG (120, 60, 30 μg/L) or simvastatin (100 μg/L). ICAM-1/VCAM-1 mRNA expression was determined by RT-PCR and protein expression was detected by immunohistochemistry and/or western blotting. The data show that ICAM-1/VCAM-1 mRNA/protein expression was significantly enhanced in atherosclerotic aortas compared with normal diet group. Ox-LDL-induced ICAM-1/VCAM-1 mRNA/protein expression in U937 cells. Importantly, TSG significantly inhibited ICAM-1/VCAM-1 expression in atherosclerotic aortas in a dose-dependent manner. TSG-pretreatment also inhibited ICAM-1/VCAM-1 expression in Ox-LDL-induced U937 cells. Therefore, we concluded that TSG suppressed the expression of adhesion (ICAM-1/VCAM-1) molecules both in vivo (in aortic wall of dietary atherosclerotic rats) and in vitro (U937 cells).
  Cell biochemistry and biophysics 04/2013;
• Article: Elevated Plasma Levels of Hypermethylated RASSF1A Gene Sequences in Pregnant Women with Intrahepatic Cholestasis.

  Ping Yi, Na Yin, Yingru Zheng, Hongmei Jiang, Xinmei Yu, Yaohua Yan, Qiang Liu, Fenglian Xiao, Li Li
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  ABSTRACT: Intrahepatic cholestasis of pregnancy (ICP) is associated with increased perinatal mortality and morbidity. Circulating cell-free fetal DNA has been a useful parameter for monitoring of pregnancy-associated diseases. The purpose of this study was to determine the concentrations of hypermethylated RAS-association domain family 1, isoform A (RASSF1A) gene sequences in the plasma of pregnant women with intrahepatic cholestasis. This study included 56 women in their third trimester of pregnancy, of whom 26 had ICP (study group) and 30 were healthy (control group). Real time PCR was performed to detect RASSF1A concentrations after methylation-sensitive restriction digestion with HinpII and HhaI to measure cell-free fetal DNA. Beta-actin was detected as an internal control to confirm complete enzyme digestion. The data show a significant increase in the circulating hypermethylated RASSF1A levels regarding the pregnancies complicated with ICP as compared with normal pregnancies. Circulating hypermethylated RASSF1A levels in maternal plasma related to total bile acid. Based on these observations, we suggest that the circulating hypermethylated RASSF1A levels in maternal plasma may be used as a diagnostic marker for ICP.
  Cell biochemistry and biophysics 04/2013;
• Article: HOXA9 Gene Expression in Acute Myeloid Leukemia.

  De-Peng Li, Zhen-Yu Li, Wei Sang, Hai Cheng, Xiu-Ying Pan, Kai-Lin Xu
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  ABSTRACT: Homeobox genes encode the class of transcription factors in vertebrates and are found in clusters called A, B, C, and D on four separate chromosomes. HOXA9 gene is part of the cluster A on chromosome 7 and encodes a DNA-binding transcription factor which may regulate gene expression, morphogenesis, and differentiation. The objective of this study was to determine the HOXA9 gene expression in acute myeloid leukemia (AML). For this purpose, semi-quantitative reverse transcriptase-polymerase chain reaction was used to measure HOXA9 gene expression in human erythroleukemia (HEL) cell line and bone marrow mononuclear cells from 54 AML patients and 20 healthy individuals. The data show that HOXA9 mRNA expression was negative in 20 healthy individuals but was positive in HEL cells and in 22 out of 54 (40.74 %) AML patients. The complete remission rate (45.45 %) of the patients who expressed the gene was significantly lower than that (71.86 %) in patients who did not express the gene after chemotherapy. Therefore, it was concluded that HOXA9 gene might be involved in the pathogenesis of AML and played as a worse prognostic factor in AML.
  Cell biochemistry and biophysics 04/2013;
• Article: Influence of Magnetic Field on Brain Activity During Administration of Caffeine.

  M I El Gohary, A A Salama, A A El Saeid, T M El Sayed, H Sh Kotb
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  ABSTRACT: The aim of the present work is to evaluate the effect of caffeine, the world's most popular psychoactive drug, on the electric activity of the rat's brain that exposed to extremely low-frequency magnetic field (ELF-MF), during 15 days. The obtained results showed that administration of caffeine in a group of rats by dose of 10 mg/kg (equivalent to human daily consumption) caused a reduction in the mean power amplitude of electroencephalogram (EEG) trace for almost all frequency bands especially α (8-12 Hz). It was observed that the influence of caffeine was more evident in motor cortex than in visual cortex. While the exposure of another group to ELF-MF of intensity 0.2 mT during the same period caused an enhancement in the mean power amplitude of most EEG frequency bands; this was more observed in the right hemisphere of the brain than that of the left hemisphere. The administration of caffeine while rats were exposed to ELF-MF, led, after 5 days of exposure, to a great increase in the mean power amplitude of α band at all places of recording electrodes. It may be concluded that caffeine administration was more effective in reducing the hazardous of ELF-MF in motor cortex than in visual cortex.
  Cell biochemistry and biophysics 04/2013;
• Article: Kinetics of Apoptosis and Expression of Apoptosis-Related Proteins in Rat CA3 Hippocampus Cells After Experimental Diffuse Brain Injury.

  Jianliang Chen, Xuesong Li, Jiandong Qiu, Hengxing You, Qingping Zhang, Guangyu Dong, You Zuo
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  ABSTRACT: The present study examined kinetics of apoptosis and expression of apoptosis-related proteins Bcl-2, Bax, and caspase-3 in the CA3 hippocampus cells after diffuse brain injury (DBI) induced experimentally in rats. Percentage of apoptotic cells and expressions of above proteins were examined by flow cytometry and immunohistochemistry. Substantial neuronal apoptosis was documented in the CA3 hippocampus cells after DBI (22.26 ± 2.97 % at 72 h after DBI vs. 2.92 ± 0.88 % in sham-operated animals). Expression of Bc1-2 decreased, while expression of Bax and caspase-3 increased after DBI, with caspase-3 expression peaking after that of Bax (72 vs. 48 h, respectively). Further, the Bc1-2/Bax expression ratio decreased prior to increase of caspase-3 expression. In conclusion, cell apoptosis and altered expressions of Bcl-2, Bax, and caspase-3 are present in the CA3 region of hippocampus after experimental DBI. Changes in the Bc1-2/Bax expression ratio may facilitate activation of caspase-3 and aggravate neuronal apoptosis after brain injury.
  Cell biochemistry and biophysics 04/2013;