rAdinbitor was cloned from Gloydius blomhoffi brevicaudus in the laboratory. Previous researches had proved that rAdinbitor could inhibit proliferation of C6 glioma cells as well as promote their apoptosis. The molecular mechanism of rAdinbitor’s effects on C6 cells need to be further studied. rAdinbitor was expressed in E. coli BL21/pET23b-adinbitor and purified with Ni Sepharose 6 Fast Flow. The purified protein was confirmed by Western blotting. C6 cells were induced with fibronectin (FN). The effects of rAdinbitor with different concentrations on the expression of FAK, MEK1/2 and Caspase-3 as well as on activity of FAK and ERK1/2 in FN-induced C6 cells were studied by immunoblotting and immunoprecipitation. Results showed that rAdinbitor with different concentrations could obviously reduce the expression of FAK and MEK1/2, increase the expression of Caspase-3, as well as decrease ERK1/2 phosphorylation; besides 10 mg/L rAdinbitor, other concentrations’ rAdinbitor could inhibit FAK phosphorylation obviously. All those effects were dose-dependent. Results indicate that the effects of rAdinbitor on decreasing expression and activity of FAK and inhibiting Ras-MAPK signaling pathway play an important role in suppressing the proliferation of C6. Furthermore, the increase in Caspase-3 expression implies that the increase in apoptosis of C6 cells might be due to the suppression of rAdinbitor on the activity of ILK and PI-3K/Akt pathway.

Objective To study the releasing properties and mechanism in vitro of the active ingredients of the gastric floating sustained-release tablet containning Zuojin Pellet extraction(ZJ-GFST).Methods The release rates of berberine,palmatine,evodiamine,and rutaecarpine from ZJ-GFST in vitro within 8 h were measured by using rotating basket method in Chinese Pharmacopeia.The cumulative curve of drug release data was fitted to zero order,first-order and Higuchi equation to ascertain the kinetic modeling of drug release.Release mechanism was ascertained using Peppas equation.Results The similar factors of the cumulative release curve of all the four ingredients mutually compared were more than 80%,indicating that the release of the four ingredients were similar.The cumulative release rate of all the four ingredients fitted Higuchi equation.The value of slopes of Peppas models of all the four ingredients were more than 0.45,indicating that drug released by concurrent action of diffusion and matrix erode(non-fickian diffusion).Conclusion The releasing properties in vitro of the active ingredients of ZJ-GFST is consistent.

Objective To study the quality standard of Folium Nelumbinis.Methods Flavonoids of Folium Nelumbinis were identified by TLC.The extraction conditions of flavonoids was studied and the content of total flavonoids was determined by ultraviolet spectrophotometry .Results Quercetin in Folium Nelumbinis was identified by TLC.The optimal extraction conditions were defined and the content of total flavonoids was about 13 %.Conclusion This method is simple and convenient, and is feasible for the quantitative determination of Folium Nelumbinis.

AIM: To preparate brucine liposome. METHODS: Brucine liposomes were preparated by a combination of ammonium sulfate transmembrane gradients method and extrusion through microfiltration membranes.The methods were evaluated by particle morphology,the size range,and encapsulation efficiency. RESULTS: The optimal preparating process parameters of brucine nanometer liposome were lecithin-to-cholesterin ratio of 100∶15,the volume of 0.1 mol/L of water solution of ammonium sulfate being 66.7 times as large as lecithin,the volume of brucine being 0.0167 times than lecithin,the temperature and the time of incubating being 40 ℃ and 20 min,respectively.The encapsulation efficiency of this method was over 90%. CONCLUSION: Preparation of brucine liposomes by a combination of ammonium sulfate transmembrane gradients method and extrusion through microfiltration membranes is feasible.

Arg-Gly-Asp (RGD) is a unique minimal integrin-binding sequence found within several glycoprotein ligands and also in snake-venom disintegrins. Adinbitor, a protein with 73 amino acid residues including 12 cysteins and an RGD motif, was cloned from Gloydius blomhoffi brevicaudus in my laboratory. As a new member of disintegrin family, adinbitor can inhibit both human platelet aggregation induced by ADP and angiogenensis in vivo and in vitro, the typical characters of disintegrin family. To separate the effect of inhibiting platelet aggregation from that of inhibiting angiogenensis, the motif KGD was introduced into adinbitor cDNA to replace RGD by site-directed and PCR-based mutagenesis. The recombinant protein (recombinant adinbitor (KGD)) was expressed in E. coli BL21 and purified through the His· Bind affinity chromatography. Recombinant adinbitor (KGD) could inhibit ADP-induced platelet aggregation with IC50 value of 85 nmol/L. Considerably, it was a more effective inhibitor on platelet aggregation than recombinant adinbitor (RGD), which has an IC50 of 150 nmol/L. Interestingly, recombinant adinbitor (KGD) has no potency in inhibiting angiogenesis in vivo compared with recombinant adinbitor (RGD). These findings showed that KGD containing adinbitor was more suitable for inhibiting ADP-induced human platelet aggregation as a potential and specific inhibitor of human platelet aggregation, which might have promising therapeutic potential as an antithrombotic agent.

Objective To analyze the failure rate and failure factors related to ventilator weaning off in children. Methods Clinical data of 214 patients who received mechanical ventilation in pediatric intensive care unit of Tianjin Children's Hospital from Jan 2005 to Dec 2008 were analyzed retrospectively. Results There were 141 planned extubation events,122 of which were successful and 19 of which were unsuccessful.The failure rate of planned extubation was 13.5%. The failure rate of infants less than 6 months was higher than that of infants more than 6 months, but there was no significant difference ( 15.0% vs 9. 8%, P ＞0.05). The success rate of extubation in pneumonia cases (90. 7% )was higher than that in others (P ＜0. 05 ). The duration of mechanical ventilation had no effect on success rate of extubation ( (7. 64 ± 5.68 ) d vs (6. 95 ± 3. 14) d, P ＞ 0. 05 ). The duration of corticosteroid treatment after extubation in the successful group was less than that in the failure group ( ( 12. 35 ±9. 69) h vs ( 18.63 ± 12. 17) h,P ＜0. 05). According to the result of multiple linear regression analysis ( R2 = 0. 093), airway obstruction was the high risk factor of the extubation failure( F = 14. 256, P ＜ 0. 001 ). Conclusion Currently,weaning from mechanical ventilation in PICU depends on the combination of pediatricians' experience and objective indicators. The key point for improving the success rate of exubation is that we should explore reasonable and feasible extubation protocol and discover and exclude the related factors of exubation failure as earlier as possible.

Objective To evaluate the diagnostic utilities of CD64,CDllb,sICAM-1 and sE-selectin in early identification of neonatal sepsis related to bacterial infection. Methods The group of sepsis consisted of 36 newboms and the control group included 26 healthy newboms. The blood samples were collected right after being admitted to hospital and at recovery stage in the group of sepsis,as for the control the blood samples were collected only once in the study. In the sepsis group,blood samples were also taken in bacterial culture before treatment CD64 and CDllb were quantified with direct immunofluorescence staining and the whole blood cell flow cytometry analysis. sICAM-1 and sE-selectin level were determined by ELJSA assay,along with CRP. Results The expression level of CD64 in neonates with sepsis was (60. 37 22. 70) .shown in MFI,which was significantly higher than that in the group of control (23. 14 ±5. 10) MFI(P <0. 01). The expression level of CDllb in neonates with sepsis was (1645. 14 ±463. 68) MFI,which was significantly higher than that in the group of control (1041.48 ±260. 34) MFI (P < 0. 01). The concentration of blood sICAM-1 in neonates with sepsis was (240. 20 ± 83.46) μg/L, which was significantly higher than that in the group of control (100. 24 ±51.03)μg/L(P <0. 01). The concentration of blood sE-selectin in neonates with sepsis was (29. 63 ±9. 88μg/L,which was significantly higher than that in the group of control (14. 12 ±5. 33)μg/L(P <0. 01). In the sepsis group,the level of CD64,CDllb,sICAM-l and sE-selectin in the primary stage was higher than the recovery stage significantly (P <0. 01). The sensitivity of above-mentioned molecular markers were 95. 7% , 82. 6% , 81. 8% and 87. 0% , respectively, and the specificity were 95. 8% , 79. 2% ,86.9% and 79.2% . CD64 was the best one. Conclusions CD64 may serve as one of the reliable biomarkers in the early diagnosis of neonatal sepsis, and it may play important role in the treatment of neonatal sepsis.

Objective:To establish fusion PCR for amplification of the full-length cDNA of dengue virus type 2. Methods:According to the published nucleotide sequence of D2-43,the primers were devised and the 5′ and 3′ half genomic cDNAs of dengue virus type 2 were amplified by long reverse transcription PCR. Using the PCR products as model,the approximate 11 kb full-length cDNA was amplified by fusion PCR. The sequence containing the 5′ noncoding region was determined by PRISMTM ABI 377 automated sequencer.Results:Using fusion PCR,the full-length cDNA of dengue virus type 2 was successfully amplified and its correctness was proved by partial nucleotide sequences analysis. To our best knowledge, this is the first report of the same kind.Conclusion:Fusion PCR is an effective method to amplify the genomic cDNA of dengue virus.

AIM: To study preparation of strychnine solid liposome. METHODS: Sorbitol carrier aggradation method and freeze drying method were used to prepare strychnine solid liposomes. They were evaluated by particle morphology, the size range, resolvable peculiarity in the water, encapsulation efficiency. RESULTS: The particles of liposome by sorbitol carrier aggradation method were less than that by freeze drying method, and its encapsulation efficiency was higher than that by freeze drying method. CONCLUSION: Sorbitol carrier aggradation method is better than freeze drying method.

ABSTRACTOBJECTIVE To observe the effect of active components from Lycii cortex on the proliferation of high glucose?in-duced glomerular mesangial cells GMCs and the secretion of extracellular matrix ECM.To investigate the protective mechanism of diabetic nephropathy.METHODS GMCs were cultured and divided into high glucose groupfinal glucose con-centration of 30 mmol/Lblank groupfinal glucose concentration of 5.5 mmol/Lbetaine and kaempferol groups of 5 dif-ferent concentrations of 0.010.1110100 μmol/L.The proliferation of high glucose?induced GMCs were observed by MTT method and secretion of ECM was detected by ELISA assay.RESULTS Different doses of betaine and kaempferol inhib-ited the proliferation of GMCsand decreased the content of ECMcompared with high glucose group P < 0.05 ~ 0.01. CONCLUSION Betaine and kaempferol can protect the high glucose?induced GMCs through inhibiting the proliferation of GMCs and decreasing the content of ECM.