Objective Nine VEGF siRNAs were designed and their efficiencys were evaluated for introducing new and effective design rules of siRNA.Methods Nine VEGF siRNA sequences with nineteen nucleic acids were designed.After construction in vitro,VEGF siRNAs were transferred into neuroglioma cells by lipofectamine.The level of VEGF protein in the media was determined by ELISA 48 hours after transferring.The relationship between the characteristics of VEGF siRNA and its' interfering effect were analyzed.Results A quantitative parameter called H-b index were used to characterize the secondary structure of VEGF mRNA.A low value of the H-b index would mean that the structure of mRNA is loose,in which the nucleotides of the targeting region are likely to be accessible by siRNA and the effects are better.The experiments showed a negative correlation between the suppression of VEGF protein expression and H-b index(r=-0.498).The H-b indexs of siRNAs targeting VEGF mRNA 386～406 and 401～421 position were less,however the effects of suppression were better.The siRNAs conferred a strong RNAi activity with A residue at position 6 and A/U residues at position 19 of sense strand of siRNAs.Thethermodynamic properties of siRNA play a critical role in the transferring effects.The lower average of internal stability values for positions 9～14 of the antisense strand is facilitating the interfering effect.The lower binding energy is the more stable incorporation between antisense strand of siRNA and mRNA present,the more favorable is for RISC cleaving mRNA.Conclusions The sequences of VEGF siRNA have been designed successfully and effectively inhibited the expression of VEGF protein.

AIM: To determine the inhibition of VEGF small interfering RNA (siRNA) on expression of VEGF protein and SMMC 7721 cell growth. METHODS: Nine VEGFsiRNA sequences with nineteen nucleotides were designed under the assistance of computer. The inhibitory effect of VEGFsiRNA was analyzed by CCK8. The protein level of VEGF in the media was determined by ELISA. The change of cell cycle was detected by flow cytometry. RESULTS: All the VEGFsiRNA were capable of inhibiting the proliferation of SMMC 7721 cells significantly compared with control and lipofectamine control, while the inhibitory effects of VEGFsiRNA2, VEGFsiRNA4, VEGFsiRNA6, VEGFsiRNA7, VEGFsiRNA8 and VEGFsiRNA9 were better than that of antisense oligodeoxynucleotide. All the VEGFsiRNA reduced the expression of VEGF protein. The effect of VEGFsiRNA7 and VEGFsiRNA2 were the best with the inhibitory rates of 52.65% and 50.43%, respectively. VEGFsiRNAs induced the S arrest in SMMC 7721 cells. The signs of apoptosis in SMMC 7721 cells induced by VEGFsiRNA were not observed. CONCLUSION: VEGFsiRNA sequences were designed and synthesized successfully. VEGFsiRNA effectively inhibited the proliferation of SMMC 7721 cells and reduced the level of VEGF protein.

AIM: To construct eukaryotic expression vector of human vascular endothelial growth factor (VEGF) gene, and to explore the effects of VEGF gene on proliferation and chemosensitivity of leukemia cells. METHODS: The recombinant eukaryotic express plasmid pEGFP-C1-hVEGF_(165) was constructed with conventional gene engineering methods. The pEGFP-C1-hVEGF_165 was transfected into leukemia cell K562. The proliferation of the recombinant eukaryotic expression plasmid was analyzed by CCK8 method. The level of VEGF mRNA was detected by reverse transcription and quantitative polymerase chain reaction (RT-PCR). The expression level of VEGF protein was determined by ELISA in cell culture medium and cells. The cell cycle was detected by flow cytometry. RESULTS: VEGF_(165) eukaryotic expression vector was successfully constructed, confirmed by enzyme digestion and sequencing. Compared with the K562 cells transfected pEGFP-C1, the K562 cells transfected with pEGFP-C1-hVEGF_(165) grew faster and expressions of VEGF mRNA and protein increased significantly. In addition, K562 cells transfected with pEGFP-C1-hVEGF_(165) had relatively higher cell survival rate to chemotherapy drugs homoharringtonine(HHT) and fluorouracil(FU). CONCLUSION: VEGF_(165) eukaryotic expression vector increases the level of VEGF mRNA and protein expression, accordingly promotes the proliferation of leukemia cells and decreases the sensitivity of leukemia cells to HHT and FU.

Objective To investigate the clinical significances of thyroid function changes in patients with post-stroke depression(PSD). Methods Sixty two cases with acute cerebral infarction (CI) were stratified into two groups by Zung self-rating depression scale, single cerebral infarct patients group (CI group) and PSD group. Serum levels of FT3, FT4 and TSH of the patients were detected on 2 days,7days,14 days and 21days after admission. Results ①On each time point after admission, FT3 levels in PSD group were lower than those in CI group. FT3 levels in the two groups on the 7 days were higher than those on 2 days(2.04±0.53 vs 1.28±0.19, 1.43±0.91 vs 1.07±0.37, P <0.05).On the 14 and 21 days, levels of FT3 in the PSD group were continuously increased.②On the 2 days and 7 days after admission, FT4 levels in PSD group were higher than those in CI group. FT4 levels in the two groups on the 14 days were lower than those on 7 days(1.52±0.18 vs 2.09±0.26, 1.57±0.31 vs 2.28±0.38, P <0.05). ③On the 2 days after admission,the TSH level in the PSD group were lower than that in CI group(2.02±0.36 vs 2.27±0.49,P <0.05). Conclusion The results indicated that the hypothalamic-pituitary-thyroid axis plays important role in the pathogenesis of PSD.

Objective To investigate the relationship between gliosis, synaptic reorganization and the mechanism of chronic temporal lobe epilepsy induced by kainic acid. Methods The mossy fibers sprouting, synaptic reorganization and gliosis were observed by Neo-Timm staining and immunohistochemical staining respectively. Results The injected (ipslateral) side was injured predominantly in the CA 3 subfield and hilus, whereas the CA 1 subfield was injured mildlyThe contralateral side showed pathological changes mainly in the CA 1 subfield and hilus, and moderately in the CA 3 subfieldThe longer the rats survived, the more obvious changes were observed in the hippocampuses. Conclusion The injection of kainic acid in hippocampus can cause neuronal loss, reactive gliosis and plasticity of neurons, probably form the abnormal neuronal discharge circuit, finally induce the seizures

Objective This study aimed to study the effect of intra-brachial nitroglycerin administration via the angiography catheter on radial artery spasm in transradial percutaneous coronary intervention. Methods From patients undergoing transradial percutaneous coronary intervention (TRI) and succeeding in coronary angiography (CAG), 810 patients who needed percutaneous coronary intervention (PCI) (including Percutaneous Transluminal Coronary Angioplasty and/or stent implantation) were erolled, including 413 patients in the study group and 397 patients in the control group. When coronary angiography completed, positioned the angiography catheter over brachial artery and patients in the group received an administration of 400μg nitroglycerin via the angiography catheter. The control group received an administration of 400μg nitroglycerin via the sheath. The incidence of RAS were recored and compared. Resultes Baseline characteristics were similar between two groups. In the study group, the incidence of RAS was signiifcatively lower (18 vs. 33, P=0.021). Conclusions Intra-brachial nitroglycerin administration via the angiography catheter can prevent RAS during the insertion of guiding catheter in TRI.

OBJECTIVE: To establish an HPLC method for the determination of rebamipide in rats' plasma and colonic tissue.METHODS: The rats were assigned to receive 4 mg rebamipide chitosan capsule or its gelatin capsule or its CMC-Na solution peros.Then the plasma and colon tissue samples were taken at different time for a determination of the concentration of rebamipide by HPLC gradient elution with Cmax computed.RESULTS: The linear range for rebamipide was from 20～2 000 ng?mL-1.The average extracting recoveries of rebamipide from the plasma and the colon were 88.4% and 90.8%,respectively.After oral administration of 4 mg rebamipide chitosan capsule or its gelatin capsule or its CMC-Na solution,the Cmax of rebamipide in the colon tissue were 5 963.9,2 190.7 and 1 185.8 ng?g-1 respectively versus 252.7,949.0 and 1 023.3 ng?mL-1 respectively in plasma.CONCLUSIONS: The established HPLC gradient elution in the study can be used to determine quantitatively the rebamipide level in plasma or colon tissue for the development of its colon specific delivery preparations.

Objective To measure the mRNA expression of immediate-early gene c-fos in endothelial cells by the use of different intermittent hypoxia (IH) protocols. Methods A gas control delivery system was homemade, which pro-duced IH/re-oxygenation(ROX) environmental exposure. The endothelial cells were exposed to IH/ROX cycles, and were di-vided into three groups (A, B and C). There were five sub-groups in each group. Group A included intermittent normoxia group, standard incubator control group, continuous hypoxia group (CH), 1.5%O2 IH group and 10%O2 IH group. There were different frequency IH of 1.5%O2 and 10%O2 sub-groups in group B and group C. Hypoxia time was 15 s. The intermittent hypoxia cycle was 60 and the ROX time was different. The total cycle time was different, including 1.5 h group, 3 h group, 5 h group, 6.5 h group and 9.5 h group. The mRNA expression of immediate-early gene c-fos was measured by real-time PCR. Results There was no significant difference in the level of c-fos mRNA between intermittent normoxia group and CH group. The expression of c-fos mRNA was significantly higher in 1.5%O2 IH group and 10%O2 IH group than that of intermit-tent normoxia group, and there was a higher expression level of c-fos mRNA in 1.5%O2 IH group than that of 10%O2 IH group (P＜0.01). It was found that the mRNA expression of c-fos increased gradually at first, and then gradually decreased. The expression of c-fos mRNA was significantly higher in 5 h group than that of other groups (P＜0.01). Conclusion The mRNA expression of immediate-early gene c-fos is increased after exposing to IH/ROX in endothelial cells, which is closely related with IH extents and frequencies.

OBJECTIVE To suvey the mastering of knowledge on prevention and treatment of AIDS among medical staff and evaluate the effect of health education on them.METHODS Suvey among medical staff was performed by self-designed questionnaires before and after education,then evaluated the effect of health education.RESULTS The mean score before education was 20.12?2.28;The mean score after education was 20.77?4.04.There were significant differences between them(t=6.417,P

OBJECTIVE:To observe the clinical efficacy of Yiru tiaojing granules on hyperprolacfinemia(HPRL)of renal deficiency and liver stagnation type.METHODS:HPRL patients were randomized to treatment group(Yiru tiaojing granules)or treatment group(Bromocriptine)for 3 months.Patients' symptoms and sex hormone were recorded.RESULTS:As compared with before treatment,serum level of prolactin(PRL)was significantly reduced(P