AIM: The effective antisense sequences targeted VEGF mRNA with computer software would be screened and designed, and effect of them on growth K562 cells and protein expression of VEGF were studied with experiments. METHODS: Seven antisense sequences were selected and synthesized, which consisted of 18-20 deoxynucleotide acid and were modified with phosphorothioate, according to principle of low free energy of overall △G 37 Overall. Cell growth was assayed by trypan blue dye exclusion assay and level of VEGF protien in the media was determined by ELISA. RESULTS: Six of seven sequences were capable of inhibing growth of K562 cells and downregulating the VEGF protein expression significantly, compared with Scrambed control group. It was found that there was a close correlation between low level of overall △G 37 and antisense effectiveness ( r =0 887, P

AIM: To investigate inhibition of K562 cell growth by antisense drug targeted VEGF mRNA. METHODS: X7, 20-mer antisense sequences were selected, synthesized and modified with phosphorothioate. The drug was transfected into K562 cells in the present of lipofection. Cell growth was assayed by trypan blue dye exclusion assay and MTT. The level of VEGF protein in the media was determined by ELISA. The morphology of apoptotic cells were observed by Giemsa staining, and the propotion of apoptotic cells was detected by flow cytometry. RESULTS: The antisense drug inhibited growth of K562 and downregulated expression of VEGF protein significantly, compared with Scrambed control group and showed dose-dependent relation. Signs of apoptosis of K562 cells were not observed. CONCLUSION: Inhibition of K562 cell proliferation, but not cells apoptosis induction is the mechanism of inhibing growth of K562 cells by antisense drug targeted VEGF mRNA. At same time, VEGF has function of promoting K562 cell proliferation, and VEGF mRNA may be a new target attached by drugs. [

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Objective To investigate the influence of abaI expression on acinetobacter baumannii biofilm formation.Methods Acinetobacter baumannii strain S isolated from bums patients was collected for the study,while the standard strain ATCC19606 was served as the control.At 6,24 and 48 hours,the gene expressions of abaI,pgaA,pgaB and pgaC were detected by real-time fluorescent quantitative PT-PCR,secretion of N-acyl-homoserine lactones (AHLs) by biological sensor and biofilm formation by MTT method.Results (1) Gene expressions of abaI,pgaA,pgaB and pgaC at 6 hours were 8.63 ±5.93,1.98 ± 1.93,1.01 ± 1.32 and 2.67 ± 3.46 respectively,which showed a quick increase at 24 hours (22.81 ± 17.60,5.13 ± 4.32,5.66 ± 3.97,11.97 ± 7.75 respectively),followed by a rapid decline in 48 hours (3.43 ± 0.88,1.30 ± 0.24,3.01 ± 3.00,3.02 ± 3.29 respectively).Gene expressions of pgaB and pgaC at 6 hours and that of pgaA and pgaC at 48 hours revealed statistically significant differences from those at 24 hours (P ＜ 0.05).(2) AHLs showed a level of 18.49 ± 11.03 at 6 hours,reached a peak of 52.23 ± 15.95 at 24 hours,then descended to 5.53 ± 0.94 at 48 hours.AHLs level at 24 hours showed statistically significant difference from that at 6 hours and 48 hours (P ＜ 0.05).(3)Biofilm formation at 24 hours and 48 hours was 2.83 ±0.44 and 2.71 ±0.15 respectively,far higher than that at 6 hours (0.49 ± 0.11,P ＜ 0.05).(4) In the correlation analysis among AHLs,biofilm formation and gene abaI,pgaA,pgaB and pgaC expressions,significant positive correlation was found between abaI and pgaA and between AHLs and pgaC expression (P ＜ 0.05).Conclusion Acinetobacter baumannii may regulate gene expressions of pgaA and pgaC responsible for biofilm formation to adjust to the external environment by means of changing abaI gene expression and AHLs secretion.

The study on mycoflora in bottled purified d rinking water was carried out.91 Samples of products were colleted from 59 bottl ing factories in Fujian Province and were examined and identified.461 Strains of fungi were isolated from 58(63 74%)different samples.Fungi imperfecti was the e umycetes isolated most frequently.These fungi were not closely related to the ae robic bacterial count and coliform of the studied waters but was clearly associa ted with the packaging of the products.The result confirmed that the cross conta minations on the process of post-purification were the main source of fungi in the final products.

Objective To investigate the relationship between proteinuria components and the severity of pregnancy induced hypertension syndrome (PIH), the unconcentrated urine samples from patients with PIH were analyzed on proteinuria components by electrophoresis.Methods Proteinuria components were analyzed by sodium dodecyl sulfate-agarose gel electrophoresis (SDS-AGE) in unconcentrated urine samples from PIH patients (PIH group,n=114) and normal third trimester pregnant women (control group,n=110).Results Eleven kinds of urinary protein were detected in the PIH group and four in the control group. The results showed positive relationship between the urine protein component complexity and the severity of PIH (P

Objective To explore radiographic and MRI features of the hemangiomas and vascular malformation in the deep soft-tissue of the extremities.Methods 89 cases confirmed by surgical pathology and angiography were analyzed retrospectively with clinical data and radiological findings.Imaging exams included plain X-ray performed in all patients,and MRI scan in 33 patients.Results On X-ray plain film,there was normal bone and soft tissue in 54 cases(60.7%),soft tissue abnormalities in 14 cases(1 5.7%),and phleboliths in 30 cases(33.7%).Also,plain X-ray film studies demonstrated bone changes adjacent to the deep soft tissue in 32 cases(36.0%), including periosteal reactions(13 cases),cortical erosion (1 6 cases),involvement of the bone marrow (10 cases)and 7 diffuse lesions with all above changes.On MRI,lesions were similar to honeycomb or sponge,and T1 WI showed isohypointense signal in 25 cases (75.8%),hypointense in 5 cases (1 5.2%),inhomogeneous slightly hyperintense in 3 cases (9.0%).On T2 WI,all lesions were well defined and showed hyperintense signals with hypointense septation,of which there were 9 cases with nodular hypointense areas and vascular flow effect.Hypointense phleboliths were showed in 10 cases(30.3%).Following injection of the contrast medium,all lesions had heterogeneous enhancement patterns.Of the 33 patients studies with MRI,lesions of 18 cases(54.5%)had bone changes which were adjacent to or partially or fully wrapped by neighbouring soft tissue lesions,in which lesions of 12 cases showed abnormal signal within bone marrow and lesions of 3 cases with vascular flow void phenomenon.In 15 cases (45.5%)without osseous change,lesions of 3 cases were adjacent to bone,and lesions of 12 cases were with fat and muscle septum between the bone and lesions.Conclusion The hemangiomas and vascular malformation in the deep soft-tissue of the extremities may cause changes in adjacent bones.Familiarity with the performance of reactive bone changes on X-ray and MRI may help to improve the diagnosis and avoid misdiagnosis.

OBJECTIVETo explore the effects of antisense phosphorothioate oligodeoxynucleotides (AS PS-ODN) of vascular endothelial growth factor (VEGF) on drug-sensitivity of AML and CML cells to homoharringtonine and its possible mechanism.

METHODSA7, which was the most effective AS PS-ODN selected by computer aid-designing and experimental assay, contains 20-mer modified with phosphorothioate. It was transferred into cells by lipofectin while cultured in 2% serum medium for 8 h and then in 10% serum medium. Twenty four hours later, homoharringtonine was added into the culture and cultured for another 48 h. Cell viability was detected by trypan blue exclusion every 24 hours, cell apoptosis by flow cytometry, and level of VEGF protein by a VEGF ELISA kit.

RESULTSThe combination of A7 and homoharringtonine was able to inhibit cell survival (AML cell survival reduced 38%, CML cell survival reduced 25%), down-regulate VEGF protein expression (VEGF protein expression of AML cell and CML cell declined 25.44% and 30.81%, respectively) and increase homoharringtonine induced apoptosis in both AML and CML cells (apoptotic rates of AML and CML cells increased 48.29% and 52.76%, respectively).

CONCLUSIONThe VEGF AS PS-ODN is able to enhance the drug-sensitivity to homoharringtonine, suggesting that inner VEGF proteins in myeloid leukemia cells had a drug-resistant effect.

Acute Disease ; Adult ; Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; genetics ; physiology ; Cell Survival ; drug effects ; genetics ; physiology ; Drug Screening Assays, Antitumor ; methods ; Female ; Harringtonines ; pharmacology ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; pathology ; Leukemia, Myeloid ; genetics ; pathology ; Male ; Middle Aged ; Oligonucleotides, Antisense ; genetics ; Transfection ; methods ; Tumor Cells, Cultured ; Vascular Endothelial Growth Factor A ; genetics ; metabolism ; physiology ; Young Adult

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