Objective To investigate the effect of mouse double minute 2 (MDM2) mRNA ASON and mismatched oligonucleotide (ASONM) radiolabeled with 99Tcm on target gene expression in LNCaP cells.Methods The ASON and ASONM targeted to MDM2 mRNA were synthesized and radiolabeled by 99Tcm with the bifunctional chelator of HYNIC.The labeling efficiency,radiochemical purity,stability and molecular hybridization activity were investigated.The different concentrations of 99Tcm-HYNIC-ASON (0,100,500 nmol/L) and 99Tcm-HYNIC-ASONM (500 nmol/L) coated with lipofectamin 2000 were incubated with prostate cancer cells for 24 h,then RT-PCR and Western blot were carried out to assay the MDM2,p53 mRNA and the corresponding protein level.The variables of RT-PCR and Western blot were analyzed using one-way analysis of variance and q test.Results The labeling efficiency of ASON and ASONM were (65.15± 2.05)％ (n=5) and (64.93±2.18)％ (n=5),respectively.The radiochemical purity were both more than 90％.99Tcm-HYNIC-ASON had a good stability and could hybridize to the sense oligonucleotide (SON).The contents of MDM2 mRNA in 0,100,500 nmol/L 99Tcm-HYNIC-ASON and 500 nmol/L 99Tcm-HYNIC-ASONM groups were 0.458±0.035,0.250±0.026,0.174±0.032,0.463±0.033,respectively,and there were significant differences between each 2 groups except between 0 nmol/L 99Tcm-HYNIC-ASON and 500 nmol/L 99Tcm-HYNIC-ASONM groups (F=33.69,q =24.32-91.45,all P＜0.01).The average density of MDM2 protein in the 4 groups were 90.712±3.042,71.218±2.915,32.775±3.062,88.121±2.710,respectively (F=235.93,q=6.43-19.14,all P＜0.01; except 0 nmol/L99Tcm-HYNIC-ASON and 500 nmol/L 99Tcm-HYNIC-ASONM).The contents of p53 mRNA in the 4 groups were 0.185±0.046,0.203±0.040,0.213±0.027,0.163±0.049,respectively(F =2.18,P＞ 0.05).The average density of p53 protein was 33.865 ± 2.213,70.445±2.180,99.025±3.012,38.351±3.271,respectively (F=53.98,q =3.32-6.74,all P＜0.01 ; except 0 nmol/L 99Tcm-HYNIC-ASON and 500 nmol/L 99Tcm-HYNIC-ASONM).Conclusions The MDM2 antisense probe can accumulate in the prostate cancer cells,and specially hybridize to the MDM2 mRNA and inhibit target gene expression.This novel molecular probe has a promising potential for the diagnosis of prostate cancer at gene level.

Objeetive To explore the value of antisense imaging of 99Tcm-labeled ASON targeting mouse double minute 2(MDM2) mRNA for the diagnosis of human prostate cancer.Methods The ASON targeting MDM2 mRNA and the mismatched oligonucleotide (ASONM) were synthesized and radiolabeled with 99Tcm using the bifunctional chelator HYNIC.The labeling efficiency and radiochemical purity were investigated.Animal models of nude mice bearing human prostate cancer LNCaP were established and divided into 3 groups with 10 mice in each group.99Tcm-HYNIC-ASON,99Tcm-HYNIC-ASONM (study groups) and 99TcmO4-(control group) were injected at the dose of 7.4 MBq through the tail vein,respectively.Tumor imaging was acquired with SPECT and the tumor-to-muscle (T/M) ratio was measured.The data was compared by one-way analysis of variance.Results The labeling efficiencies of ASON and ASONM were (65.15± 2.05) ％ and (64.93±2.18) ％,respectively.Their radiochemical purity was greater than 90％.At 1,4 and 10 h post injection,the T/M ratios of 99Tcm-HYNIC-ASON group were 3.217±0.125,3.749± 0.201 and 4.028±0.186,and those of 99Tcm-HYNIC-ASONM group were 1.579t0.128,1.715±1.140 and 1.683±0.139,and control group 2.146±0.132,1.847±0.124,1.528±0.152,respectively.The T/M ratios in control group and 99Tcm-HYNIC-ASONM group were significantly lower than those in 99Tcm-HYNICASON group at 1,4 and 10 h,respectively (F=213.37-235.41,t=3.527-4.738; all P＜0.01).The T/M ratios of 99Tcm-HYNIC-ASONM group and control group were not significantly different at 1,4 and 10 h (t=2.154,2.287 and 2.236,all P＞0.05).Conclusion The antisense probe of MDM2 can accumulate specifically in prostate cancer tissue in animal models,which might be useful as a non-invasive genetic tool for the early diagnosis of prostate cancer.

Objective:To optimize the formula of 5-aminosalicylic acid enema in situ gel. Methods:5-Aminosalicylic acid ene-mas in situ gel was prepared using a cold dissolution method with carbomer as the gel matrix and xanthan gum as the thickener. A 32 full factorial design was used to investigate the effects of the concentrations of carbomer and xanthan gum on the viscosity before and af-ter the gelling, duration of inversion tube and sedimentation rate. Response surface methodology was used to optimize the formula. Re-sults:The quantitative relationships between the two factors and the four evaluation indices were obtained. The optimum formula was as follows:the concentration of carbopol and xanthan gum in the enema was 0. 7% and 0. 15%, respectively. The viscosity before and af-ter the gelling was 500-1 000 mPa·s and 2 200-2 700 mPa·s, respectively. The duration of inversion tube test was 40-80 min and the sedimentation rate was more than 98. 5%. Conclusion:The multi-objective simultaneous optimization of the formula of 5-aminosal-icylic acid enema in situ gel is accomplished by factorial design and response surface methodology.

Objective To explore the protective effects of dipeptidyl peptidase-4 inhibitor (DPP-4I) on AD-like neurodegenerative changes and its mechanism. Methods The human neuroblastoma cell line SH-SY5Y on the logarithmic phase was divided into six groups:control group (CON group, treated with PBS contained 1‰DMSO for 12 h), wortmannin intervention group (W group, treated with 0.03 μmol/L wortmannin for 12 h), DPP-4I intervention group (DPP-4I group, treated with 10μmol/L DPP-4I for 12 h), both DPP-4I and wortmannin intervention group (DPP-4I+W group, pre-treated with 10 μmol/L DPP-4I for 2 h, then 0.03 μmol/L wortmannin for 12 h), DPP-4I, wortmannin and Ex9-39 intervention group (DPP-4I+W+Ex9-39 group, pre-treated with 10μmol/L Ex9-39 for 2 h, then 10μmol/L DPP-4I for 2 h followed by 0.03μmol/L wortmannin for 12 h), and Ex9-39 intervention group (Ex9-39 group, treated with 10μmol/L Ex9-39 for 12 h). MTT assay was used to detect the cell vitality. Western blot assay was used to detect the level of total tau protein (tau-5) and phosphorylated tau at different sites (pSpS199/202, pT231 and pS396), the level of phosphorylated neurofilaments (NF-H, NF-M) and phosphorylation of critical enzyme in PI3K/Akt/GSK-3β signaling pathway. Results (1) The cell vitality decreased, the levels of pSpS199/202, pT231, pS396 and NF-H/M increased significantly in W group than those in CON group. However, comparing with CON group, the above mentioned parameters reversed in DPP-4I group. Comparing with W group, the cell vitality increased and phosphorylated levels of above mentioned indices were decreased in DPP-4I+W group. (2) The cell vitality showed a decline trend while the levels of phosphorylation tau at three different sites and NF-H/M were higher in Ex9-39 group than those in CON group. Comparing with DPP-4I+W group, the results of the phosphorylated levels showed the same changes in DPP-4I+W+Ex9-39 group. (3) Comparing with CON group, the expression levels of phosphorylated PI3K, Akt and GSK3β increased significantly in DPP-4I group, while those decreased in W group. Additionally, the expression levels of phosphorylated PI3K, Akt and GSK3β were significantly increased in DPP-4I+W group than those in W group. Conclusion DPP-4I can enhance the level of GLP-1 and activate PI3K/Akt/GSK-3βinsulin signaling pathway to improve the hyperphosphorylated tau and NFs induced by wortmannin, and to protect AD-like neurodegeneration.

To explore the changes in metabolites in the greasy tongue coating in patients with chronic gastritis.

Objective To evaluate the detection rate and the efficiency of the modified crushing-cercariae escaping method.Methods The detection rates of the modified crushing-cercariae escaping method and the crushing methods were compared by using a double-blind control experiment with the latter as a gold standard.meanwhile the number of the cercariae was quantified.The efficiency of the two methods aforementioned and the cereariae escaping method were compared in field.Results The detection rate of the modified crushing-cercariae escaping method was 100%.the average number of cercariae in each infected snail was (4 778±1 157);and the number in certain volume of water sample was positively correlated with the number of infected snails.The efficiency of the modified menthed Was 18.2 times and 17.3 times as high as those of the crushing method and cereariae escaping method,respectively.Conclusions The modified crushing-cercariae escaping method Can detect the infected snails quicky and Can quantify the number of infected snails and cercariae,and is suitable for the detection of infected snails in large number.

ObjectiveTo investigate the practicability of gastric intubation with zebra-guidewire under the induction of anesthesia with sevoflurane (zebra-guidewire was the obsolete guide wire in percutaneous nephrolithotomy in urology surgery). MethodsThe patients who needed gastric intubation were randomized into the experimental group and the control group with 20 patients in each group.the experimental group received gastric intubation with zebra- guidewire under the induction of anesthesia with sevoflurane.In the control group,the gastric tubes were catheterized with routine method.The physiological indices changes,incidence rate of adverse reaction and success rate of one-time catheterization and average time of catheterization per patient were compared between the two groups. ResultsIn patients of the experimental group,the blood pressure,heart rate before and during the intubation did not fluctuate evidently,but in the control group the results were the opposite.In the experimental group the incidence rate of adverse reaction,success rate of one-time catheterization and average time of catheterization per patient were superior to the control group. ConclusionsGastric intubation under the induction of anesthesia with sevoflurane was a painless way of gastric intubation.Gastric intubation with zebra-guidewire under anesthesia state was simple and fast.

OBJECTIVE To investigate the nuclear proteomes in mitochondrial DNA (mtDNA)-depleted A549 cells (Rho0 cells) and their parental cells (Rho+ cells),and to learn more about the nuclear responses to mitochondrial dysfunction.METHODS The nuclear proteomes of Rho and Rho + cells were characterized by two dimensional electrophoresis (2-DE) and SELDI-TOF ProteinChip technologies,the differentially expressed protein- spots were identified by MALDI-TOF mass spectrum (MS),the nucleophosmin and P53 expression were detected by Western blotting assay,and the mitochondrial memhrane potential (MMP) was measured by the laser scanning confocal microscope.RESULTS 2-DE results showed 11 protein-spots were down-regulated and 21 protein-spots were up-regulated in Rho0 cell nuclei.SELDI-TOF MS analysis with NP20 ProteinChips revealed 4 protein-peaks decreased in Rho0 cell nuclei.One down-regulated protein-spot was identified as elF-6,and 4 up-regulated proteinspots were identified as nucleophosmin,SFRS1,SFRS3 and hnRNP G,respectively.The increased expression of nucleophosmin in Rho0 cells was verified by Western blotting.Based on the clues from proteomic analysis,P53 expression in Rho0 cells was higher than in Rho + cells,and MMP was consistent in Rho + and Rho0 cells.CONCLUSION mtDNA-depletion induces nuclear proteome alteration.Rho0 cells can be used as a model to study the crosstalk between mitochondrion and nucleus.

ObjectiveTo assess the characteristics of left ventrlcular ( LV) systolic syncnrony before and 6 days after percutaneous transluminal septal myocardial ablation(PTSMA)in patients with hypertrophic obstructive cardiomyopathy (HOCM) using two dimensional speckle tracking imaging(2DSTI),and to investigate correlation between the LV systolic synchrony and LV outflow-trace gradient (LVOTG).MethodsLVOTG were measured before and 6 days after PTSMA in 21 patients with HOCM.The two dimensional loop-cinec were obtained in three levels of the short axis views of LV (mitral valve,papillary muscle and cardiac apex).The time from the onset of QRS complexes to systolic peak strain from the radial vectors (TRs) was recorded using 2D-STI.The standard deviation of the TRS of 18 segments (TRsSD) were calculated as indicator of LV systolic synchrony.Results Six days after PTSMA,LVOTG decreased significantly from (84.3 ± 19.1)mm Hg to (40.6 ± 8.3)mm Hg( P ＜0.01).TRS of the base and middle segments of anteroseptal were prolonged dramatically( P ＜0.05) 6 days after PTSMA compared with that before.There were no significantly differences in TRs-SD of LV between before and 6 days after PTSMA ( t =1.03,P ＞ 0.05).Decreased LVOTG correlated moderately to prolonged TRS of the base segments of anteroseptal ( r =0.657,P ＜0.05).Conclusions2D-STI can detect exactly the characteristics of LV systolic synchrony before and 6 days after PTSMA in patients with HOCM.

Objective In order to know the effect of heating infusion for premature infants to pro-mote their restoration of physiological function. Methods Divided 160 premature infants who need infu-sion into the experimental group and the control group randomly, there were 80 cases in each group. Normal temperature infusion was used in the control group, while heating infusion was used in the experimental group, compared the body temperature, pluse, respiration, blood pressure, oxygen saturation and the blood ghcose between the two groups. Results There were significant difference about the indexes which had mentioned above between the two groups. Conclusions Heating infusion can effective promote the reha-bilitation of physiological function of premature infants.