STI571 is a new antileukemia agent targeting BCR-ABL tyrosine kinase and successfully used in treatment of leukemia patients. Despite strong efficacy of the tyrosine kinase inhibitor STI571, resistance has been observed in a significant proportion of leukemia patients. This review focuses on the diverse mechanisms of ST1571 resistance and its preventive measures.
Animals ; Antineoplastic Agents ; pharmacology ; Benzamides ; Drug Resistance, Neoplasm ; Humans ; Imatinib Mesylate ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; drug therapy ; Piperazines ; pharmacology ; Pyrimidines ; pharmacology

ObjectiveTo identify serum specific protein biomarkers for the early diagnosis of lymphoma patients by using surface enhanced laser desorption/ionization time of flight mass spectrometry (SELDI-TOF-MS)protein chip array.MethodsSELDI-TOF-MS technique and weak cation exchanger (WCX2)protein chip were used to detect the serum proteomic patterns of 96 patients with lymphoma (86 patients with non-Hodgkin lymphoma and 10 patients with Hodgkin lymphoma)and 30 normal control.The serum level of LDH were measured by biochemical methods,the serum level of β2-MG and CA125 were detected by enzyme-linked immunosorbent assay (ELISA).ResultsFour protein peaks pattern mass/charge ratio (M/Z) 6880,8564,8692 and 13751 were decreased in lymphoma patients,and they could be used to distinguish lymphoma from healthy people and other malignant tumors.Their sensitivity and specificity were 82.29 ％ and 80.00 ％,78.13 ％ and 73.30 ％,75.00 ％ and 80.00 ％,71.88 ％ and 83.30 ％ respectively in lymphoma patients.When it was used to early diagnosing lymphoma in stage Ⅰ or Ⅱ,the sensitivity of four protein peaks pattern (M/Z 6880,8564,8692 and 13 751) were 77.55 ％,71.43 ％,71.43 ％ and 67.35 ％respectively and the specificity of all four protein peaks pattern were 100.00 ％.Four protein peaks pattern sensitivity were higher than LDH,CA125 and β2-MG respectively.ConclusionApplication of SELDI-TOF-MS can be of great potential for the early diagnosis of lymphoma patients.

Pure culture of Phlebopus portentosus was inoculated in the roots of coffee tree. The results indi-cated that the young fruit bodies would come out around the rhizomes of host tree after inoculation in 30 to 90 days, single or cluster, 3 to 4 days for mature, weight 20.0 g to 62.0 g. Brown rhizomorph and hyphae can be seen on the seedlings`rhizome, main root and side root while nothing is on the tip of the root.It was found that rhizomorph on the surface of roots would die after inoculation in 90 days in pot.

OBJECTIVETo investigate the effect of hypoxia on isolated human periodontal ligament stem cells (hPDLSC) in vitro.

METHODShPDLSC were exposed to normoxia (20% O(2)) and hypoxia (1.5% - 2% O(2)). Total cellular RNA and protein were collected on day 1, 2, 4 and 6 after culture. Western analysis and semi-quantitative RT-PCR were used to analyze osteogenic differentiation of hPDLSC, including alkaline phosphatase (ALP), osteocalcin (OCN), secreted protein acidic and rich in cysteine (SPARC) and bone morphogenetic protein-2 (BMP-2).

RESULTSIn the first two days hypoxia slightly increased the growth of hPDLSC [A value was 0.697(hypoxia) vs 0.617 (nomoxia)] and after the third day hypoxia dramatically decreased the growth of the hPDLSC [A value was 0.870(hypoxia) vs 1.242 (nomoxia)]. Up to 90% reduction of ALP activity was observed in hPDLSC after 5 days of hypoxia [A value was 0.004(hypoxia) vs 0.049(nomoxia)]. Hypoxia decreased SPARC expression at protein level and down-regulated ALP, OCN and BMP-2 expression at mRNA level in comparison with nomoxia.

CONCLUSIONSHypoxia inhibited proliferation of hPDLSC and down-regulated BMP-2 mRNA expression, then down-regulated their target genes such as ALP, OCN, and SPARC, thus inhibiting critical steps in osteogenic differentiation of hPDLSC.

Bone Morphogenetic Protein 2 ; metabolism ; Cell Differentiation ; Cell Hypoxia ; Cells, Cultured ; Humans ; Periodontal Ligament ; cytology ; metabolism ; Stem Cells ; cytology ; metabolism

OBJECTIVETo test the bovine cementoblasts (CBs) cementum-forming ability in vivo.

METHODSRoot fragments of newborn bovine freshly extracted mandibular incisor were cultured routinely and 4th-5th passages of CBs were harvested. CBs were then cultured in the medium supplemented with 50 mg/L alpha-ascorbic acid and 10 mmol/l beta-glycerolphosphate to form a thick layer as tissue engineering scaffold for cementum formation. Collagen membrane was used as control scaffold. 2 x 10(6) cells were attached to the CBs-made carrier as well as collagen membrane scaffolds and transplanted subcutaneously into immunodeficient mice. Transplants were harvested at 7th week. Histological sections were stained with HE, alizarin red S and van Kossa methods as well as monoclonal Ab against bovine cementum attachment protein (CAP).

RESULTSCBs-made scaffold supported more cementum-like tissue (CLT) formation than collagen-made scaffold. The CLT formed on CBs scaffold was partly calcified with embedded cells. Uncalcified cementoid-like material could be seen on the surface and was encircled by cubical CB-like cells. The CLT was also positive to CAP and van Kossa staining.

CONCLUSIONSThese results suggest that the bovine CBs can form cementum-like tissue. The cell-made carrier is a better scaffold than collagen membrane.

Alkaline Phosphatase ; analysis ; Animals ; Bone Transplantation ; methods ; Cattle ; Cell Adhesion Molecules ; analysis ; Cells, Cultured ; Dental Cementum ; chemistry ; cytology ; transplantation ; Immunohistochemistry ; Integrin-Binding Sialoprotein ; Male ; Mice ; Mice, Nude ; Osteocalcin ; analysis ; Osteonectin ; analysis ; Sialoglycoproteins ; analysis ; Tissue Engineering ; methods ; Transplantation, Heterologous

OBJECTIVETo investigate the repression of c-myc induced by Blimp-1 in 2-methoxyestradiol(2ME2)-mediated differentiation of multiple myeloma cells.

METHODSCZ-1 and LP-1 myeloma cells lines were exposed to 0.5 micromol/L of 2ME2 and 0.5 micromol/L of 2ME2 + antisense oligonucleotides (ASODN) for 72 h. The effects on transcription of c-myc mRNA were studied by RT-PCR. The c-myc protein was assayed with Western blot. The changes of the cell lines in morphology, expression of surface CD49e and quantity of immunoglobin light chain secretion in the supernatant were studied.

RESULTSIncubation of the cells with 0.5 micromol/L of 2ME2 could up-regulated Blimp-1 expression and increase the expression of c-myc gene. In contrast, c-myc expression was decreased with Blimp-1 expression down-regulated by ASODN and the cell lines differentiation was arrested.

CONCLUSIONBlimp-1 could directly repress the expression of c-myc in 2ME2-mediated differentiation induction of multiple myeloma cells.

Cell Differentiation ; drug effects ; genetics ; Cell Line, Tumor ; Estradiol ; analogs & derivatives ; pharmacology ; Gene Expression Regulation, Neoplastic ; Humans ; Multiple Myeloma ; genetics ; metabolism ; pathology ; Positive Regulatory Domain I-Binding Factor 1 ; Proto-Oncogene Proteins c-myc ; genetics ; metabolism ; RNA, Messenger ; genetics ; Repressor Proteins ; metabolism ; Transcription Factors ; genetics ; metabolism

Objective To study the effect of different concentrations of fluoride on cultured human umbilical vein vascular endothelial cells(HUVEC). Methods Different doses of sodium fluoride (NaF) were added to HUVEC culture medium, fluoride concentrations were 0(control), 100,400,700,1000,2000 μmol/L, respectively,6 re-set hole in each group. After continuous culture for 48 h, cells and culture medium were collected. Cell morphology was studied by Wright-Giemsa staining; cells apoptosis was determined by acridine orange fluorescence staining; cell activity was measured by methyl thiazolyl tetrazolium (MTT) assay; superoxide dismutase (SOD),glutathione peroxidase(GSH-Px) activity, malonaldehyde(MDA) content, induced nitricoxide synthase(iNOS), and endothelia nitricoxide synthase(eNOS) activity in cell culture medium were determined by spectrophotometry; cell iNOS mRNA and eNOS mRNA expression were detected by RT-PCR; intercellular adhesion molecule-1 (ICAM-1)and vascular cell adhesion molecule-1 (VCAM-1) levels were detected by double antibody sandwich ELISA method.Results With increased dose of fluoride, HUVEC cells decreased, the structure changed. In 400 - 2000 μmol/L group, the SOD activity[(6.627 ± 0.213), (6.668 ± 0.152), (5.935 ± 0.122), (4.755 ± 0.182)kU/L] was lower than those of the control group[(7.457 ± 0.398)kU/L, P ＜ 0.05 or ＜ 0.01], GSH-Px activity[(481.284 ± 43.785),(492.223 ± 16.474), (382.762 ± 25.167), (293.687 ± 24.881 )kU/L] was also lower than those of the control group [(585.078 ± 47.323)kU/L, P ＜ 0.05 or ＜ 0.01], MDA level[(0.609 ± 0.011 ), (0.646 ± 0.016), (0.852 ± 0.013),(1.188 ± 0.045)nmol/L] was higher than those of the control group[(0.512 ± 0.027)nmol/L, P ＜ 0.05 or ＜ 0.01];iNOS activity[(3.604 ± 0.115), (3.615 ± 0.075), (3.848 ± 0.103), (4.275 ± 0.079)kU/L] also was higher than those of the control group[(2.798 ± 0. 136)kU/L, all P ＜ 0.01], iNOS mRNA expression increased, eNOS activity [(5.539 ± 0.079), (5.503 ± 0.064), (5.226 ± 0.142), (4.809 ± 0. 107)kU/L] decreased compared to those of control group[(5.996 ± 0.155)kU/L, P ＜ 0.05 or ＜ 0.01], eNOS mRNA expression decreased; ICAM-1 levels [(0.852 ± 0. 102), (0.886 ± 0.061 ), (0.961 ± 0.158), (1.418 ± 0. 167)μg/L] increased compared to those of the control group[(0.687 ± 0.046)μg/L, P ＜ 0.05 or ＜ 0.01], VCAM-1 levels[(2.719 ± 0.197), (2.946 ± 0.167),(3.173 ± 0.225 ), (3.613 ± 0. 153 ) μg/L] was higher than those of the control group [(2.375 ± 0.067 ) μg/L, all P ＜0.01]. Conclusions High concentrations of fluoride reduce the activity of antioxidant enzymes, which leads to metabolic disorders of nitric oxide and abnormal cytokines expression, thereby inhibiting vascular endothelial cell growth, structural change and induced apoptosis. This is an important factor in high fluoride-induced vascular endothelial injury.

OBJECTIVETo study the protective effect of formula of removing both phlegm and blood stasis (TYTZ) on myocardial tissues of Chinese mini-swine with coronary heart disease of phlegm-stasis cementation syndrome.

METHODTotally 36 Chinese mini-swine were randomly divided to six groups: the normal control group, the model group, the Danlou tablet group, and TYTZ groups with doses of 2.0, 1.0, 0.5 g x kg(-1), with six in each group. Except for the normal control group, all of other groups were fed with high-fat diet for 2 weeks. Interventional balloons are adopted to injure their left anterior descending artery endothelium. After the operation, they were fed with high-fat diet for 8 weeks to prepare the coronary heart disease model of phlegm-stasis cementation syndrome in Chinese mini-swine. After the operation, they were administered with drugs for 8 weeks. The SOD activity and MDA content of each group were observed at the 0th week (before the experiment), the 2nd week after the high-fat diet (before the operation or drug administration) , the 6th week after the high-fat diet (4 weeks after the drug administration) and the 10th week after the high-fat diet (8 weeks after the drug administration). Meanwhile, the myocardial enzymogram test and the HE staining pathological observation were performed at the end of the experiment. The changes in the myocardial cell ultra-structure were observed under transmission electron microscope.

RESULTCompared with the normal control group, the model group showed significant decrease in serum SOD activity and notable increase in MDA content from the 2nd week to the end of experiment (P < 0.05 and P < 0.01). In the 10th week, the CK, LDH and CK-MB levels in serum also significantly increased in the model group (P < 0.05 and P < 0.01), with obvious structural abnormality in myocardial tissue pathologic morphology and ultra-structure. Compared with the model group, TYTZ groups showed specific increase in serum SOD activity and oblivious decrease in the MDA level (P < 0.05 or P < 0.01). Meanwhile, TYTZ could significantly decrease serum CK and LDH levels in the model group (P < 0.05 or P < 0.01), attenuate the ischemia injury of myocardial tissue, and improve the ultra-structure of cardiomyocytes.

CONCLUSIONTYTZ shows an obvious protective effect on the myocardial injury in Chinese mini-swine with coronary heart disease of phlegm-stasis cementation syndrome. Its mechanism is related to the resistance against free radical oxidation injury and the inhibition of the lipid per-oxidation.

Animals ; Coronary Artery Disease ; genetics ; metabolism ; prevention & control ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Humans ; Male ; Mucus ; drug effects ; metabolism ; Myocardium ; metabolism ; Protective Agents ; administration & dosage ; Swine ; Swine, Miniature

OBJECTIVEDevelopment and application of a real time fluorescent quantitative PCR (FQ-PCR) assay for detecting WU polyomavirus in children with low respiratory tract infections.

METHODSThe VP2 gene of WU polyomavirus was selected as the detection target, from which the real time primers and probes were designed. The standard curve was established by using recombinant plasmid as template. And the FQ-PCR assay for specific detection of WU polyomavirus was established. The specificity, sensitivity and reproducibility of the method were evaluated. Furthermore, the clinical specimens from children with respiratory tract infections collected in Wenling First People's Hospital were quantitatively detected using this method.

RESULTSIn this study, the FQ-PCR method was established to detect a specific fragment in VP2gene of WU polyomavirus. The standard curve coefficient R2 was 0.998. And this method can detect as low as 50 copies recombinant plasmid. The clinical specimens of sputum and throat swab from children with respiratory tract infections were quantitatively detected using this method. 7 sputum specimens were detected as WU polyomavirus positive in 700 sputum specimens, the positive ratio was 1.00%. No positive specimens were detected in 146 specimens of throat swabs and 846 blood samples from same patient population.

CONCLUSIONThe results indicated that the FQ-PCR assay method established in this study was specific, rapid and sensitive for detecting WU polyomavirus in children with lower respiratory tract infections. The sputum specimen is more suitable to be used for gene detection of WU polyomavirus than throat swab or blood.

Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Polyomavirus ; isolation & purification ; Real-Time Polymerase Chain Reaction ; methods ; Respiratory Tract Infections ; virology ; Sputum ; virology

BACKGROUNDIn our previous studies, we found the expression of 14-kD phosphohistidine phosphatase (PHPT1) was associated with lung cancer cells migration and invasion, and PHPT1 mRNA expression level in lung cancer tissues clinically correlated with lymph node metastasis. In the present study, we aimed to further investigate the expression of PHPT1 protein in lung cancer.

METHODSExpression of PHPT1 protein in tissue samples from 146 lung cancers and 30 normal tissues adjacent to lung cancers was assessed using immunohistochemical method. Fisher's exact test was used to analyze expression patterns of PHPT1 protein in these tissue types. Meanwhile, we studied the correlation between expression of PHPT1 protein and clinicopathological features in lung cancer.

RESULTSSignificantly higher expression levels of PHPT1 protein were found in lung cancer samples (53.42%) than in normal tissues adjacent to lung cancer (23.33%) (P = 0.003). Fisher's exact test showed that lung cancer stage positively correlated with expression of PHPT1 protein (P = 0.02), and lung cancer samples with lymph node metastasis showed higher PHPT1 protein expression (P = 0.016) than the samples without lymph node metastasis.

CONCLUSIONSThe results of this study agree with findings from our previous study of PHPT1 mRNA expression in lung cancer tissues, and strongly suggest that PHPT1 protein is closely associated with the carcinogenesis and metastasis of lung cancer. Thus, therapy targeting PHPT1 (inhibition or silencing) could be potentially benefited for lung cancer patients.

Blotting, Western ; Gene Expression Regulation, Neoplastic ; Humans ; Immunohistochemistry ; In Vitro Techniques ; Lung Neoplasms ; metabolism ; Phosphoric Monoester Hydrolases ; metabolism