Objective To explore the association between type 2 diabetes mellitus (T2DM)combined with non -alcoholic fatty liver disease (NAFLD)and serum adiponectin (APN)and its clinical significance.Methods A total of 76 patients who were initially diagnosed with T2DM or had been diagnosed with T2DM for less than three years and admitted to the endocrine department and its outpatient clinic of the Affiliated Hospital of Medical School of Ningbo University from August 2011 to July 2013 were selected as the subjects,and were assigned to group A or B,each consisting of 38 patients,based on the presence or absence of associated NAFLD.Another 30 healthy subjects receiving physical examination were assigned to the control group (NC group).The indicators including body mass index (BMI),blood lipids,blood glucose,liver function,insulin resistance index (HOMA -IR)and serum APN,were measured and compared among the groups. Results In group A ,the indicators including BMI,triglyceride (TG),total cholesterol (TC),low density lipoprotein -cholesterol (LDL -C),alanine aminotransferase (ALT),fasting insulin (FINS)and HOMA -IR were significantly higher than those in group B and NC group,and the APN level was significantly lower than that in group B and NC group (P <0.05).Pearson correlation analysis indicated that APN was negatively correlated with BMI,TG,high density lipoprotein -cholesterol(HDL -C),and HOMA -IR,and positively correlated with TC,LDL -C and ALT(P <0.05).Multiple linear regression analysis suggested that HDL -C and HOMA -IR was the independent influencing factors for APN (P <0.05).Conclusion Severe insulin resistance and glucose and lipid metabolic disorders are present in T2DM patients combined with NAFLD,and adiponectin level is of vital significance for the development and progression of NAFLD.

Objective To study the effects of vascular endothelial growth factor(VEGF)on production of pulmonary surfactants.Method Fetal rat lungs were obtained at 19-day gestation.Primary culture of typeⅡalveolar epithelial cells(AECⅡ)was performed using IgG panning technique.The rats was divided into groups: VEGF,Dexamethasone,VEGF plus Dexamethasone and a control.Total phospholipids,phosphatidylcholine (PC),phosphatidyl glycerol(PG)and sphingornyelin(SM)were determined.Results expressed as mean?SEM. Comparison between groups were made with LSD-t test and one -way ANOVA.Result VEGF,Dexamethasone and VEGF plus Dexamethasone groups showed increased amount of total phospholipids and its compositions on the first day of culture.Conclusions VEGF-165 promotes the production and secretion of pulmonary surfactant. VEGF and Dexamethason may go through different mechanism for enhancement of synthesis of pulmonary surfactant,thereby improve biological function of AECⅡ.

Objective To study the influence of vascular endothelial growth factor(VEGF) on development of alveolar epithelial cell Ⅱ (AECⅡ) and expression of pulmonary surfactant protein B(SP-B) in premature rats.Methods Wistar rats at 19 days gestation were paunched to get embryo and primary AECⅡculture.The rats were divided into 4 groups ,VEGF-165 group,Dexamethasone group,VEGF and Dexamethason group,control group. AECⅡ and SP-B expression were measured by immunology histochemistry.Results SP-B had positive expression in VEGF group, Dexamethason group, VEGF and Dexamethason group. SP-B had negative expression in control group.Conclusion VEGF-165 can increase SP-B positive expression and secret of AECⅡ.VEGF promotes lung maturity.

Objective To investigate homografting vascular endothelial progenitor cells(EPCs)for preventing restenosis formation of carotid artery in New Zealand white rabbit models.Methods EPCs of New Zealand white rabbits were isolated,confirmed and expanded though the injured carotid arterial endothelium of rabbit model induced by dilatation with a 2.5 F balloon;and then EPCs were transplanted into the injured endothelium of the cells transplantation group(n=13,3 of them were transplanted with fluorencently-labeled- EPCs),while equal volume of saline without EPCs was injected into the injured endothelium in the control group(n=8).Histopathology was performed at 4 days after transplantation for the 2 rabbits,with fluorencently-labeled-EPCs.All of the rest remained rabbits were killed 4 weeks later for histological examinations.Results The histopathological slides showed that the fluorescence-positive expression existed in the injured endothelium 4 days after transplantation.At 4 weeks after the EPCs transplantation,there were less restenosis and less vascular wall thickening in the rabbits of cells transplantation group than those of the control group(P＜0.01).Conclusion The local interventional homografting heterogeneous endothelial progenitor cells can prevent restenosis after the carotid artery angioplasty in New Zealand White rabbit model. (J Intervent Radiol,2007,16:95-98)

BACKGROUND AND OBJECTIVEWe previously reported that C-KIT overexpression and mutation exist in biopsy samples of nasopharyngeal carcinoma (NPC). Yet whether Imatinib had an inhibitory effect on the proliferation of NPC in vitro was still unknown. So, this study examined whether sensitivities to Imatinib of other cell lines are different and whether C-KIT expression and mutations exist, to analyze the correlations between them.

METHODSThe expression of C-KIT in NPC cell lines, including CNE-1, CNE-2, Hone-1, C-666, SUNE-1, 5-8F, and nasopharyngeal epithelial (NPE) cell line NP-69, were detected by Western blot. Direct sequencing of polymerase chain reaction (PCR) products was performed to analyze the sequences of C-KIT from the above-mentioned cell lines. Inhibitory effects on proliferation by Imatinib on these cell lines were determined by CCK-8 assay. Pearson product moment correlation and t test were used to analyze the correlation betweeen C-KIT overexpression, C-KIT gene mutation, and the inhibitory effect of Imatinib.

RESULTSCompared with NPE cell line NP-69, NPC cell lines CNE-1, CNE-2, Hone-1, C-666, SUNE-1, and 5-8F had significantly higher levels of C-KIT expression. Heterozygous IVS17+78T>C were found in CNE-1, CNE-2, Hone-1, and NP-69 cell lines, homozygous IVS17+78T>C was found in C-666, and no mutation was found in SUNE-1 or 5-8F. Imatinib had a dose-dependent inhibitory effect on proliferation for CNE-1, CNE-2, Hone-1, C-666, SUNE-1, and 5-8F. No significant correlation between the inhibitory effects of Imatinib, C-KIT overexpression, or C-KIT mutation was found.

CONCLUSIONC-KIT overexpression and intron mutation were found in NPC cell lines and Imatinib had a dose-dependent inhibitory effect on proliferation for NPC cell lines, yet no significant correlation between C-KIT overexpression, C-KIT mutation, or the inhibitory effect of Imatinib was found.

Antineoplastic Agents ; pharmacology ; Benzamides ; Carcinoma, Squamous Cell ; genetics ; metabolism ; pathology ; virology ; Cell Line ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Epithelial Cells ; cytology ; metabolism ; Herpesvirus 4, Human ; isolation & purification ; Heterozygote ; Homozygote ; Humans ; Imatinib Mesylate ; Introns ; Mutation ; Nasopharyngeal Neoplasms ; genetics ; metabolism ; pathology ; virology ; Nasopharynx ; cytology ; Piperazines ; pharmacology ; Proto-Oncogene Proteins c-kit ; genetics ; metabolism ; Pyrimidines ; pharmacology

OBJECTIVETo explore the feasibility of in vitro magnetic resonance imaging on Fe2O3-arginine labeled heNOS gene modified endothelial progenitor cells (EPCs).

METHODSFe2O3 was incubated with arginine to form Fe2O3-arginine complex. Rabbit peripheral blood mononuclear cells (MNCs) were isolated and EPCs were isolated by adherence method, expanded and modified with heNOS gene using Lipofectamine 2000. After 48 hours, genetically modified EPCs were incubated with Fe2O3-arginine for 24 hours. Intracellular iron was detected by Prussian blue stain. The expression of heNOS gene was detected by Western blot. MTT assay was used to evaluate cell survival and proliferation of Fe2O3-arginine labeled heNOS-EPCs. Flow cytometry was used to measure cell apoptosis. The cells underwent in vitro MR imaging with various sequences.

RESULTSIron-containing intracytoplasmatic vesicles could be clearly observed with Prussian blue staining, and the labeling rate of labeled heNOS-EPCs were similar to that of labeled EPCs (around 100%). Survival and apoptosis rates obtained by MTT and flow cytometry analysis were similar among labeled heNOS-EPCs, labeled EPCs and unlabeled EPCs with Fe2O3-arginine. The signal intensity on MRI was equally decreased in labeled heNOS-EPCs and labeled EPCs compared with that in unlabeled cells. The percentage change in signal intensity (DeltaSI) was most significant on T2*WI and DeltaSI was significantly lower in cells labeled for 7 days than that labeled for 1 days.

CONCLUSIONSThe heNOS gene can be successfully transfected into rabbit peripheral blood EPCs using Lipofectamine2000. The heNOS-EPCs can be labeled with Fe2O3-arginine without significant change in viability and proliferation capacity. The labeled heNOS-EPCs can be imaged with standard 1.5 T MR equipment. The degree of MR signal intensity may indirectly reflect the cell count, growth and division status.

Animals ; Endothelial Cells ; cytology ; Ferric Compounds ; Humans ; In Vitro Techniques ; Magnetic Resonance Imaging ; methods ; Male ; Nitric Oxide Synthase Type III ; genetics ; Rabbits ; Stem Cells ; cytology

OBJECTIVETo investigate the changes in brain injury after the induction chemotherapy in children with acute lymphoblastic leukemia (ALL) by cranial MRI.

METHODSThe clinical data and cranial MRI results of 62 children with ALL who were hospitalized from March 2014 to June 2015 were analyzed retrospectively.

RESULTSBefore chemotherapy, MRI showed bone marrow infiltration of the skull in 33 patients (53%); the children with WBC<20×10(9)/Lhad a significantly lower incidence rate of bone marrow infiltration of the skull than those with WBC≥20×10(9)/L (16 patients/42% vs 17 patients/71%; P<0.05), and the high-risk group had a significantly higher incidence rate of bone marrow infiltration of the skull than the non-high-risk group (71% vs 44%; P<0.05). Before chemotherapy, there were 4 cases (7%) of brain atrophy, and 2 cases (3%) of abnormal signals in the sensory conduction bundle. MRI reexamination in 28 patients after 3 months of chemotherapy showed 3 new cases (11%) of brain atrophy and 1 aggravated case of brain atrophy.

CONCLUSIONSThe children with ALL have bone marrow infiltration of the skull, brain atrophy, and abnormal signals in the sensory conduction bundle before chemotherapy, especially bone marrow infiltration of the skull, and some changes in brain injury disappear after treatment.

Adolescent ; Bone Marrow ; pathology ; Brain ; drug effects ; pathology ; Child ; Child, Preschool ; Female ; Humans ; Induction Chemotherapy ; adverse effects ; Infant ; Magnetic Resonance Imaging ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; drug therapy ; pathology ; Retrospective Studies ; Skull ; pathology

Coronary artery disease (CAD)is a major cause of death and disability worldwide, and consumes a considerable amount of medical resources every year.Clopidogrel is a first-line antiplate-let therapy for CHD, butit is associated with substantial variability in PK and pharmacodynamics re-sponse. To date, gene variants explain only a smallproportion of the variability.The study aimed to identify new genetic loci-modifying antiplatelet response to clopidogrel in Chinese patients with CAD by a systematic analysis combining antiplatelet effects and PK, and further to investigate the PON1 gene promoter DNA methylation and genetic variations possibly influencing clinical outcomes in pa-tients undergoing PCI. We identified novel variants in two transporter genes (SLC14A2rs12456693, ATP-binding cassette [ABC]A1 rs2487032) and in N6AMT1 (rs2254638) associated with P2Y12 reac-tion unit (PRU) and plasma active metabolite (H4) concentration. These new variants dramatically im-proved the predictability of PRU variability to 37.7％. The associations between these loci and PK pa-rameters of clopidogrel and H4 were observed in additional patients, and its function on the activation of clopidogrel was validated in liver S9 fractions (P<0.05). Rs2254638 was further identified to exert a marginal risk effect formajor adverse cardiac events in an independent cohort.Multivariate logistic regression analysis indicated that PON1methylation level at CpG site-161 (OR=0.95; 95％ CI=0.92–0.98;P<0.01)and the use of angiotensin converting enzyme inhibitors(OR=0.48;95％ CI=0.26–0.89;P<0.01) were associated with decreased risk of bleeding events. In conclusion, new genetic variants were systematically identified as risk factors for the reduced efficacy of clopidogrel treatment.The ab-normal expression of DNA methylation-regulating key genes in the pharmacokinetic and pharmacody-namics pathways of clopidogrel and aspirin may modify clinical outcomes in dual antiplatelet-treated pa-tients undergoing PCI.

OBJECTIVETo investigate the expression of beta-catenin in patients with leukemia and explore its significance in leukemias.

METHODSRT-PCR was used to detect the expression of beta-catenin in bone marrow mononuclear cells (BMMNCs) from patients with leukemia. Immunocytochemistry was in some of patients to detect the distribution of beta-catenin at the same time. The clinical significance of beta-catenin was analyzed in combination with patients' clinical information.

RESULTSExpression of beta-catenin was statistically higher in acute myeloid leukemia (AML) and acute lymphocytic leukemia (ALL) samples than in normal donors (P = 0.001 and 0.016 respectively) and chronic phase chronic myeloid leukemia (CML) patients (P = 0.001 and P = 0.008 respectively), while there was no statistic difference between AML and ALL patients (P = 0.58). In addition, beta-catenin expression in chronic phase CML patients was like that in normal donors (P = 0.49), but increased significantly in blast crisis and accelerated phase. Immunocytochemical analysis revealed that BMMNCs from normal donors expressed beta-catenin on the plasma membrane and cytoplasma, while those from acute leukemia expressed beta-catenin to varying degrees in the nucleus as well. The expression of beta-catenin gene statistically showed the highest level in M5 (n = 15) and the lowest level in M3 (n = 18). No clinical features, such as, age, initial WBC count, therapy response rate, blast cell numbers or cytogenetic risk was found to be correlated with the expression of beta-catenin excepting for CD34+ positive rate (P = 0.004) in AML.

CONCLUSIONAs a key mediator of Wnt signal transduction way, overexpression of beta-catenin in leukemia cells indicates that it might be aberrantly activated in acute leukemia, accelerated or blastic phase of CML.

Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; Female ; Humans ; Leukemia ; metabolism ; Male ; Middle Aged ; RNA, Messenger ; metabolism ; beta Catenin ; metabolism

OBJECTIVETo inhibit the expression of beta-catenin and investigate the effect of the beta-catenin gene on Jurkat and K562 cells.

METHODSsiRNA specifically knocking down the expression of beta-catenin was used to testify the function of beta-catenin in Jurkat and K562 cells. Real time polymerase chain reaction and Western blot were performed respectively to testify the mRNA level and protein level of beta-catenin. Growth curve was determined by counting viable cells using trypan blue refusal-dyed method. The proliferation of cells was assayed by clonogenic counting and MTT method. The apoptotic cells were measured by Annexin V/PI staining. The cell cycle analysis was performed based on propidium iodide staining.

RESULTSCompared with the control group (transfected with siRNA directed against scramble gene), the survival, colonogenicity, and proliferation of the Jurkat and K562 cells were significantly decreased in experimental group transfected with beta-catenin siRNA. The colonogenicity was decreased from 31.9 +/- 5.55 (siRNA) to 25.0 +/- 5.13 (control) in Jurkat cells, and from 47.33 +/- 8.52 (siRNA) to 39.33 +/- 6.26 (control) in K562 cells (both P <0.05). The inhibition rate was (49.3 +/- 9.86)% (siRNA) and (15.1 +/- 6.55)% (control) respectively in Jurkat cells, and (39.4 +/- 7.56)% (siRNA) and (10.1 +/- 6.89)% (control) in K562 cells (both P <0.05). In addition, the apoptotic rate increased from (23.5 +/- 2.82)% (control group) to (55.9 +/- 2.22)% (experiment group) in Jurkat cells and from (14.9 +/- 8.54)% (control group) to (27.9 +/- 15.3)% (experiment group) in K562 cells. However, cell cycle analysis revealed no obvious phases change both in Jurkat and in K562 cells.

CONCLUSIONKnock-down of beta-catenin gene may decrease the proliferation, survival, and clonogenicity in Jurkat cells and K562 cells.

Apoptosis ; genetics ; physiology ; Blotting, Western ; Cell Cycle ; genetics ; physiology ; Cell Line ; Cell Proliferation ; Humans ; Jurkat Cells ; cytology ; metabolism ; K562 Cells ; cytology ; metabolism ; RNA, Small Interfering ; genetics ; physiology ; Reverse Transcriptase Polymerase Chain Reaction ; beta Catenin ; genetics ; metabolism ; physiology