Objective To inactivate Death-associated protein kinase 1 gene (DAPK1) by transfecting complementary methylated oligonucleotides and studies its effect on the proliferation of myelogenous leukemia cell line K562. Methods Methylated oligonucleotides complementary to DAPK1 gene promoter were transfected into K562 cells by Iipo2000. Methylation specific PCR (MSP) and Reverse transcription PCR (RT-PCR) were applied to detect DAPK1 gene promoter methylation status and its mRNA expressions respectively. MTT was used to detect the proliferation of K562 cells pre- and post- oligonucleotides transfection. Results DAPK1 gene promoter in non-treated and control groups were unmethylated with detectable mRNA expressions, but it became methylated with inhibited mRNA expressions after methylated oligonucleotide transfection. Proliferation in methylated oligonucleotide treatment group was significantly higher than that in non-treated and control groups. Conclusion Complementary methylated oligonucleotides could inactivate DAPK1 gene and inhibit its expression in K562 cells, which could promote its proliferation.

Objective To investigate the characteristics of HBsAg and HBcAg in combination pulsed monocyte-derived DCs(MoDC) derived from patients with HBV-associated hepatocellular carcinoma(HCC).Methods MoDCs were generated from 20 HBV-associated HCC patients,and pulsed with recombinant human serum albumin(rHSA)as conDC(control DC),or pulsed with HBsAg andHBcAg in combination as scDC.Phenotypic patterns of MoDCs were characterized by flow cytometry,and the levels of cytokines releasedby MoDCs were analyzed by ELISA,and frequencies of IFN-γ-producing antigen-specific autologous T cells were measured by Elispotassay.HBV-specific CD8 T cells proliferation was determined by pentamer staining.Results 1.The levels of MHC and costimulatorymolecules expressed on scDC were significantly higher than those on immature MoDC(imaDC)in two groups,and the levels of MHC andcostimulatory molecules expressed on imaDC,conDC and scDC in HBsAg+,anti-HBe+,anti-HBc+patients(1-4-5 positive group)were significantly higher than those in HBsAg+,anti-HBc+patients(1-5 positive group);2.The levels of IL-12 and IL-10 produced byscDC were higher than those produced by imaDC and conDC in both groups,and the levels of IL-12 produced by imaDC,conDC and scDCin 1-4-5 positive group were higher than those in 1-5 positive group.3.The frequencies of IFN-γ-producing T cells induced by scDC werehigher than those by conDC in 1-4-5 positive group.4.scDCs from 4 cases of HLA-A2+patients in 1-4-5 positive group could induceautologous T cells to generate HBVcore18-27-specific CD8 T cells.Condusions HBsAg and HBcAg pulse in combination couldsubstantially reverse the impaired function of MoDCs in HBv-associated HCC patients,and boost MoDC to induce HBV-specific T cellsresponse,especially in HBsAg+,anti-HBe+ and anti-HBe+ patients.

Objective To have knowledge of the exact cause of massive osteolysis. Methods Hair of patients from Xinjiang province was collected and 14 trace elements were determined by inductively coupled plasma mass spectrometryICP-MS. Results Trace elements imbalance in the body of patients was disturbed. Chromium and zinc which are benefit to the growth of the bones were only 0.5 ?g/g and 40 ?g/g respectively that were much lower compared with the healthy persons cadmium was much higher than the limit level in healthy person. Moreover the quantity and ratio of potassium and sodium in the patients were obvious abnormal. Conclusion According to the result of the present paper may be the environmental and dietary factors play an important role in pathogenesis of this disease.

Objective To analyze the effect of bifrontal decompressive craniectomy on patients with refractory diffusing of brain swelling after severe traumatic brain injury.Methods The clinical data of 68 patients with refractory diffusing of brain swelling after severe traumatic brain injury were analyzed retrospectively.Thirty-five patients were performed with bifrontal decompressive craniectomy as observed group,continued intracranial pressure monitoring after surgery.Thirty-three patients were treated conservatively to reduce intracranial pressure as control group,continued intracranial pressure monitoring.The Glasgow outcome scale after discharge 6 months were assessed.The efficacy,the incidence of complications were observed in observed group.Results The admission intracranial pressure in observed group was significantly higher than that after surgery [(35.9 ±6.9) mmHg (1 mmHg =0.133 kPa) vs.(17.5 ±5.2) mmHg,P ＜0.05].The admission intracranial pressure in control group was (34.2 ± 8.6) mmHg,after admission 10.5 h was (32.0 ±4.8) mmHg (P ＜0.05),difference was no statistically significant (P＞ 0.05).The intracranial pressure after admission 10.5 h in control group was significantly higher than that in observed group after surgery (P ＜0.05).Two cases of subdural effusion,1 case of postoperative hydrocephalus in observed group.The better prognosis rate in observed group was significantly higher than that in control group [45.7％ (16/35) vs.18.2％ (6/33),P ＜ 0.05].Conclusions Bifrontal decompressive craniectomy is a suitable measure to decrease the intracranial pressure in the patients with refractory diffusing of brain swelling.If carried out early,it could provide better outcome for these patients.

OBJECTIVE To discuss a highly effective method to immobilize probe on the surfaces of piezoelectric DNA sensors.METHODS Pseudomonas aeruginosa probe was immobilized on the gold surface of gene sensor(array) with routine self-assembly method(SAM)(non-reduction method) and SAM with deoxidized probe((reduction) method),respectively.The changes in frequency and time-cost were compared in reactions with(different) concentrations of probe.RESULTS Reduction method had the advantage of more probe immobilization;less time consumed in testing and higher changes in frequency during the reaction than non-reduction method.CONCLUSIONS Reduction method has a better ability to immobilize probe on the surfaces of piezoelectric DNA sensors.

OBJECTIVE To establish a SNP detection method by DNA piezoelectric biosensor and detect a SNP relative to HBV infection. METHODS To establish a model experiment with synthesis DNA sequences as target and find the lowest sensitivity. After extraction of genome DNA from inpatient blood sample, the SNP sites located in ESR1 gene region in samples were detected by SNP detecting method established. RESULTS The frequency shift of target-A was 416.0?21.5Hz, the frequency shift of target-G was 9.4?5.0Hz. And it could be detected that the lowest sensitivity of target-A was 2?10-11 mol/L. The three genotypes of blood samples, TT, TC and CC, had different frequency shifts, 109.4?13.4Hz, 52.0?11.4Hz and 7.2?4.5Hz, respectively. CONCLUSIONS SNP in blood sample could be detected specifically and sensitively by DNA piezoelectric biosensor.

Objective To investigate the relationship of serum ferritin(SF) and acute coronary syndrome(ACS) .Methods ACS group(n= 110) was divided into unstable angina(UA) subgroup(n= 46) ,non-ST-segment elevation myocardial infarction (NSTEMI) subgroup(n=30) and ST-segment elevation myocardial infarction(STEMI) subgroup(n=34) according to clinical da-ta ,coronary angiography and electrocardiogram examination .The normal coronary patients provided as control group(n=42) .The levels of serum ferritin ,hs-CRP and serum lipids were determined and the differences of SF among the groups were analyzed .The correlation of hs-CRP ,TG ,LDL-C ,HDL-C ,TC and SF were also analyzed .Results Compared with control group ,the level of SF in UA ,NSTEMI ,STEMI were significantly increased(P<0 .05) .Comparison in the levels of SF in NSTEMI ,STEMI group and UA group had significant difference(P<0 .05) .Correlation analysis indicated that the level of SF were positively correlated with hs-CRP ,TG and negatively associated with HDL-C .Conclusion The level of SF significantly increased in patients with ACS and has a positive relationship with hs-CRP and the disorder of lipid metablism .SF helps in assessing the clinical classification and risk strati-fication in patients with ACS .

Objective There are three categories of bioaerosol laser telemetry systems according to the light source configuration parameters:namely traditional lidar(light datection and ranging,laser radar), micro-pulse lidar and pseudo-random modulation(lidar).The system source parameters,which impact the degree of danger and detection sensitivity of the system, need to be optimized.Methods With reference to the USA laser product safety standards and by establishing the corresponding mathematical model of a lidar, the three categories of lidar source configuration were compared according to signal to noise ratio(SNR) and security before the repetition rate, pulse energy, divergence angle, distance and other dangerous impact factors were calculated.Results The results showed that to ensure eyesafety, the use of the pulse frequency should be set at 55 kHz for the highest SNR under the micro-pulse lidar excitation mode.Conclusion The eyesafety requirements impact the excitation of light source of a bioaerosol telemetry system.

BACKGROUND: Vascular endothelial growth factor (VEGF) is able to effectively treat the ischemic heart disease, but in vivo VEGF cannot be maintained effective concentration. OBJECTIVE: To detect the expression of VEGF mRNA and protein in human bone marrow mensenchymal stem cells transferred by VEGF-165 gene. DESIGN, TIME AND SETTING: The empirical study was conducted from March 2006 to April 2007 at Shanghai Chest Hospital. MATERIALS: Human bone marrow mesenchymal stem cell line and VEGF were offered by Basic Laboratory, Thoracic Tumor Institute, Shanghai Chest Hospital. METHODS: hVEGF165 gene was reconstructed in pcPGK-vector and transferred into human bone marrow mensenchymal stem cells (BMSCs) by liposome-mediated method, clone screening by G418. MAIN OUTCOME MEASURES: The mRNA and protein of VEGF gene in transferred cells was detected by reverse transcription-polymerase chain reaction (RT-PCR), Real time PCR, Western Blot, enzyme linked immunosorbent assay (ELISA) in 3 stem cells of pcPGK-VEGF165-IRES-GFP and pcPGK- IRES-GFP, respectively. RESULTS: pcPGK-hVEGF165 vector was reconstructed and transferred into hMSCs successfully. The expression of hVEGF165 in the transfected hMSCs was demonstrated with RT-PCR and Real time PCR. Western Blot and ELISA demonstrated that the expression of hVEGF165 in the transfected hMSCs and VEGF protein in supernatant were significantly more than untransfected hMSCs. CONCLUSION: hVEGF165 can be successfully transfected into BMSCs by using liposome mediated gene transfer. Stably expressed VEGF165 cell line can be obtained..