AIM:To investigate the effect of RWDD3 gene silencing on the biological characteristics of human glioma U251 cells.METHODS: A lentiviral vector expressing RWDD3 shRNA was constructed and transfeeted into the U251 cells.The expression of RWDD3 at mRNA and protein levels was detected by real-time PCR and Western blot , re-spectively .The cell activity was determined by MTT assay .The colony formation ability was detected by the colony forma-tion assay .The cell proliferation ability was detected by BrdU incorporation assay .The cell invasion and migration were evaluated by Transwell assay .Flow cytometry was used to monitor the changes of cell cycle distribution and apoptosis .RE-SULTS:Recombinant lentivirus was successfully transfected into U 251 cells.Compared with the cells transfected with the scrambled shRNA and control cells, the cell activity, colony formation ability, and the invasive and migratory activities were inhibited, the cell cycle was arrested in G 0/G1 phase, and the apoptosis was increased in the U 251 cells transfected with RWDD3 shRNA ( P <0.05 ) .CONCLUSION: RWDD3 plays a vital role in proliferation and invasion of glioma cells.It may serve as a potential target of gene therapy for glioma .

Objectives To investigate characterization of imipenem resistance among imipenem non susceptible Pseudomonas aeruginosa isolated from patients who treated without imipenem and explore risk factors of imipenem resistance.Methods From April,2006 to March,2008,a total of 37 non-susceptible to imipenem without imipenem therapy isolates were collected from affiliated 3201st Hospital of Medical College of Xi'an Jiaotong University.The minimum inhibition concentration (MIC) to 11 antimicrobial agents were determined by the broth dilution method.We also tested imipenem MIC combined with efflux pump inhibitor PAβN.PCR was performed to check for the presence of carbapenem-hydrolylzing MBL genes and oprD gene.The expression level of oprD2 and ampC were evaluated by qRT-PCR.Molecular typing was performed using PFGE.Results There is significant difference ( t =- 2.9004,P ＜ 0.01 ) of the average number days of therapy between with two or more antibiotics in the 16 patients (20.0 ± 9.5 ) d and that with only one antibiotic in the other 21 patients ( 12.6 ± 4.4 ) d before imipenem-non-susceptible strains were isolated.In all 37 strains,32 strains showed resistance to more than three antibiotics.The MBL gene ( IMP-9 ) was only found in one strain,but its phenotype is negative,oprD2 gene from the 29 strains were found forward inserted by ISpa1328.Thirty-five isolated were considered to have no oprD expression.The patterns of the total DNA of 37 strains appeared six PFGE types.The 26 strains belonged to C2 PFGE type.In the presence of PAβN,all 37 strains increased sensitivity to meropenem.Conclusion Fluoroquinolones and cephalosporins treatment could play an important role in imipenem non-susceptible production in the research isolates.

OBJECTIVE To develop an HPLC method for determination of main components in prepared rhubarb decoction and investigate its effects on rats with noxious heat blood stasis syndrome.METHODS The analysis was performed on a Hy-persil GOLD C18column with the mobile phase of 0.1％phosphoric acid solution(A)-acetonitrile(B).The detection wave-length was set at 254 nm and the column temperature was 30 ℃.The method was validated and the samples were determined under this condition.The abdominal aortic blood of rats with noxious heat blood stasis syndrome was collected and tested to obtain the hemorheological parameters.Also,the TXB2and 6-keto-PGF1αlevels in rats plasma were measured.RESULTS The developed HPLC method showed good linearity within certain concentration ranges.And the precision,repeatability,sta-bility and recovery all met the requirements.Compared with rats in model group,the whole blood viscosity at four shear rates(P<0.05),plasma viscosity(P<0.01)and the level of plasma fibrinogen content(P<0.01)in the high dose group were sig-nificantly decreased after doing,and the TT(P<0.01)and PT(P<0.01)values were significantly prolonged.Furthermore,the levels of TXB2(decreased,P<0.05)and 6-keto-PGF1α(increased,P<0.05)were significantly changed,and their ratio was corrected to be normal.CONCLUSION The method was simple,accurate and reliable,which could be used for quality control of prepared rhubarb decoction.Prepared rhubarb could improve vascular endothelial function by modifying blood rheol-ogy,promoting the secretion of PGI2,reducing the generation of TXA2and recovering the balance of TXB2/6-keto-PGF1α.

Acute Disease ; Biomarkers, Tumor ; analysis ; Child ; Cyclin-Dependent Kinase Inhibitor p16 ; analysis ; Histocytochemistry ; Humans ; Leukemia ; metabolism ; pathology ; Proliferating Cell Nuclear Antigen ; analysis ; Proto-Oncogene Proteins c-bcl-2 ; analysis

Adolescent ; Bacteria ; isolation & purification ; Child ; Child, Preschool ; Humans ; Infant ; Infant, Newborn ; Respiratory Tract Infections ; microbiology

OBJECTIVETo study the biological properties of human dental pulp cells (HDPC) by cloning and analysis of genes differentially expressed in HDPC in comparison with human gingival fibroblasts (HGF).

METHODSHDPC and HGF were cultured and identified by immunocytochemistry. HPDC and HGF subtractive cDNA library was established by PCR-based modified subtractive hybridization, genes differentially expressed by HPDC were cloned, sequenced and compared to find homogeneous sequence in GenBank by BLAST.

RESULTSCloning and sequencing analysis indicate 12 genes differentially expressed were obtained, in which two were unknown genes. Among the 10 known genes, 4 were related to signal transduction, 2 were related to trans-membrane transportation (both cell membrane and nuclear membrane), and 2 were related to RNA splicing mechanisms.

CONCLUSIONThe biological properties of HPDC are determined by the differential expression of some genes and the growth and differentiation of HPDC are associated to the dynamic protein synthesis and secretion activities of the cell.

Cloning, Molecular ; Cloning, Organism ; Dental Pulp ; Fibroblasts ; Gene Library ; Gingiva ; Humans ; Polymerase Chain Reaction

OBJECTIVETo study the modulatory effect of Panax gingseng and coadministration with Veratrum nigrum on the activity and mRNA expression of cytochrome P450 isoenzymes in rat liver.

METHODRat liver microsomal cytochrome P450, b5, aminopyrine N-demethylase(APND), p-nitrophenol-hydroxylase(pNPH)activities were quantitated by UV chromatography. The mRNA expression level of five CYP isoenzymes CYP1A1, CYP2B1/2, CYP2C11, CYP2E1 and CYP3A1 were detected by semi-quantitative reverse transcriptase-polymerase chain reaction(RT-PCR).

RESULTP. gingseng coadministrated with V. nigrum obviously decreased the P450 contents of liver microsomes, and the b5 contents. Both single and combined used inhibited the activities of aminopyrine N-demethylase. At the mRNA level, the expression of CYP2C11 markedly induced exposure to V. nigrum, but combinative groups decreased the expression of CYP2C11. The combination of P. gingseng and V. nigrum induced the expression of CYP1A1. P. gingseng has inhibitory effect on CYP2B1/2 and inductive effect used with V. nigrum. The combination of P. gingseng with V. nigrum also induced the expression of CYP3A1.

CONCLUSIONP. gingseng used singly has some different modulation effects compared with combinative used, which may occur because of drug-drug interaction based on cytochrome P450. To elucidate the drug-drug interaction, it needs further analysis and metabolism research.

Aminopyrine N-Demethylase ; metabolism ; Animals ; Cytochrome P-450 Enzyme System ; biosynthesis ; genetics ; Cytochromes b5 ; metabolism ; Drug Incompatibility ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Female ; In Vitro Techniques ; Isoenzymes ; biosynthesis ; genetics ; Male ; Microsomes, Liver ; metabolism ; Panax ; chemistry ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Rats, Wistar ; Veratrum ; chemistry

OBJECTIVETo evaluate the morphological change of masseter after the mandibular angle osteotomy.

METHODSComputerized tomography (CT) examination was performed on 120 patients treated by mandibular angle osteotomy before operation and at 3, 6, 12 months after operation, respectively. The pre- and postoperative masseter muscle thickness and cross-sectional area were evaluated using 3D CT images observed from 3 selected slice planes, which were paralleled with Frankfurt horizontal plane. These CT images were stored and three-dimensional reconstruction were made for calculation of masseter muscle volume through software.

RESULTSAfter operation, the reduction of the masseter muscle volume and cross-sectional area was seen. The volume of the masseter at 3, 6, 12 months postoperatively decreased to 82.02%, 77.00% and 80.43% (P < 0.05). The cross-sectional area at 3, 6,12 months postoperatively decreased to 85.81%, 78.86% and 81.56% at A plane, 80.94%, 75.03% and 77.04% at B plane, and reached to 13.46%, 11.48% and 13.89% at C plane (P < 0.05, P < 0.05, P < 0.01). The masseter thickness after operation was significantly different from that before operation during the follow-up period, but not at 12 months after operation at A plane.

CONCLUSIONSThe masseter atrophy happens spontaneously after mandibular angle osteotomy, especially at the region of mandibular angle. It should be considered during surgical design.

Adolescent ; Adult ; Female ; Humans ; Mandible ; surgery ; Masseter Muscle ; anatomy & histology ; diagnostic imaging ; Osteotomy ; Postoperative Period ; Tomography, X-Ray Computed ; Young Adult

The aim of this study was to investigate the expression status of the helicase antigen (HAGE) transcript and its clinical significance in patients with acute myeloid leukemia (AML) and chronic myeloid leukemia (CML). The expression of HAGE cDNA in bone marrow mononuclear cells from AML and CML patients was detected by using real-time quantitative PCR. The results indicated that overexpression of HAGE transcript (117.12% - 9842.70%, median 434.96%) was detected in 14.8% (11/74) AML patients. AML patients with HAGE cDNA expression were significantly older than those HAGE-negative patients (median 67 and 45 years, respectively, p = 0.001). HAGE cDNA expression was more frequently present among the patients with acute monoblastic leukemia (M(4) and M(5), 7 of 20, 35.0%), compared to the patients with acute non-monoblastic leukemia (M(1), M(2), M(3) and M(6), 4 of 54, 7.4%) (p = 0.007). 28.6% (8/28) cases with normal karyotypes showed HAGE cDNA overexpression, significantly higher than 7.5% (3 of 40) in those with chromosomal abnormalities (p = 0.041). Overexpression of HAGE transcript was found in 9 (34.6%) CML cases and more frequently observed at accelerated phase and blast crisis (4/4, 100%) than that at chronic phase (5/22, 22.7%) (p = 0.008). It is concluded that HAGE cDNA expression is relevant to specific subtypes of AML and to the progression of CML.
Blast Crisis ; Bone Marrow Cells ; metabolism ; DEAD-box RNA Helicases ; genetics ; metabolism ; DNA, Complementary ; Gene Expression ; Humans ; Karyotype ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; metabolism ; Leukemia, Myeloid, Acute ; genetics ; metabolism ; Neoplasm Proteins ; genetics ; metabolism ; RNA, Messenger ; genetics

OBJECTIVETo discuss the effect of Yisui Shengxue granules on expression of alpha-hemoglobin stabilizing protein (AHSP) mRNA in different developmental stages mice.

METHODThe total RNAs were extracted from the bone marrow karyocyte of normal adult mice and the karyocyte of fetus liver and fetus spleen in pregnanted mice (pregnanted 21 days) and fetal mice (pregnanted 14 days). The expression level of AHSP mRNA in different developmental stages mice interfered with Yisui Shengxue granules was measured by real-time PCR.

RESULTThe intervention of Yisui Shengxue granules could significantly up-regulated the expression levels of AHSP mRNA in normal adult mice.

CONCLUSIONThe result revealed that one of possible molecular mechanism of the effects caused by Yisui Shengxue granules is that it can promote the AHSP gene expression, reduce the free a-globin deposit, then prevent the poison to erythrocyte and decrease the haemolysis.

Animals ; Blood Proteins ; genetics ; Bone Marrow Cells ; cytology ; drug effects ; metabolism ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Erythrocytes ; cytology ; drug effects ; metabolism ; Female ; Gene Expression Regulation, Developmental ; drug effects ; Liver ; cytology ; embryology ; metabolism ; Male ; Mice ; Molecular Chaperones ; genetics ; Plants, Medicinal ; chemistry ; Pregnancy ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Spleen ; cytology ; embryology ; metabolism ; Up-Regulation ; drug effects ; genetics