BACKGROUND: The ulcer wound is hard to heal in diabetic patients,and it is believed to be caused by the microcirculatory disorder of wound and decreased contents of endogenous growth factors in patients with diabetes mellitus.OBJECTIVE: To observe the biological effects of adeno-associated virus (AAV) mediated transforming growth factor beta1 (AAV-TGFβ1) and vascular endothelial growth factor (AAV-VEGF) in promoting the dermal ulcer healing of diabetic rabbits.DESIGN: A randomized controlled animal experiment.SETTINGS: Medical College, Qingdao University; Affiliated Hospital of Medical College, Qingdao University.MATERIALS: The experiments were carried out in the gynecological laboratory, Affiliated Hospital of Medical College, Qingdao University from July 2004 to January 2006. Twenty-four healthy adult New Zealand rabbits were randomly divided into co-transfection group (n=12) and control group (n=12).METHODS: ① The dermal ulcer models of diabetic rabbits was established by injecting alloxan (130 mg/kg) via ear vein, and the ulcer wound was made by operation. ② In the co-transfection group, the wound was locally infiltrated, and injected with AAV-TGFβ1 virus and AAV-VEGF virus (the concentration was 9×106 virus granules/mL respectively). The rabbits in the control group were treated with injection of saline.MAIN OUTCOME MEASURES: ① The levels of TGFβ1 and VEGF gene transcription in the healing tissue were detected with polymerase chain reaction (PCR) at 1 month postoperatively. ② The capillary density in the wound margin was counted with microcirculation microscope at 3 weeks postoperatively. ③ The collagen Ⅰ and Ⅲ were isolated and detected with Western blotting by protein gel electrophoresis and semi-dry electrophoretic transfer. ④ The content of collagen in the ulcer healing issue was detected at 1 day, 1, 2 and 3 weeks, 1, 2, 3, 4, 5 and 6 months postoperatively. ⑤ The thickness, color and quality of the wound healing were observed postoperatively.RESULTS: All the 24 rabbits were involved in the analysis of results. ① The expressions of TGFβ1 and VEGF gene transcription in the co-transfection group were increased as compared with those in the control group. ② The capillary density at the wound margin in the co-translection group was higher than that in the control group [(19.18±3.56), (6.43±1.52)/mm2,t=3.21, P ＜ 0.05]. ③ The content of collagen Ⅰ in the ulcer healing tissue in the co-transfection group was obviously increased, and the proportion of collagen Ⅰ in the constituent ratio of collagen Ⅱ and Ⅲ was increased.④ The contents of collagen in the ulcer healing tissue at 1 day after transfection were close without significant difference between the two groups (t=1.55, P ＞ 0.05). After that, the contents of collagen in the cotransfection group were all significantly higher than those in the control group (P ＜ 0.05). ⑤ The ulcer healing in the control group was obviously lagged with bad quality.CONCLUSION: The combination of AAV-TGFβ1 and AAV-VEGF can efficiently transfect the dermal ulcer wound in diabetic rabbits, obviously increase the capillary density in ulcer tissue, markedly improve the proportion of collagen I in the constituent ratio of collagen Ⅰ and Ⅲ in healing tissue, and obviously ameliorate the condition of ulcer healing.

Objective To study the phosphorylation mode of extracellular regulated kinase( ERK)induced by acute and chronic morphine treatment on Chinese hamster ovary( CHO)cells expressed withμopioid receptors. Methods The time course of ERK phosphorylation 1 h and 36 h after morphine exposure as well as naloxone-precipitated withdrawal was detected by immunobloting. Results A transient enhancement of ERK phosphorylation was induced by 1 μmol · L-1 morphine with the peak effect at 5 min(P〈0. 01),and the effect was dose-dependent. No difference in ERK phosphorylation was found after 36h of treatment with 10 μmol · L-1 morphine compared with the control. However,5 or 10 min-naloxone precipitation induced remarkable decrease in ERK phosphorylation compared with the control(P〈0. 01). Conclusion Different changes of ERK phosphorylation were found under acute and chronic morphine treatment and naloxone precipitation,indicating a compensation of ERK related pathway induced byμ opioid receptors.

By analyzing school factor of honesty and credit shortage in students,we put forward concrete measures to enhance honesty and credit education in students from aspects of education,management and university surroundings construction.It is expected to offer better help to honesty and credit education in colleges and universities.

Calorie restriction refers to a dietary regimen low in calories without malnutrition.During the recent 70 years,the benefit of calorie restriction regimen has been explored extensively.Work in widely diverse species,from model organism to rodents even human beings,has demonstrated that calorie restriction is an effective nutritional intervention for retarding aging and preventing chronic diseases.

OBJECTIVE:To study the in vitro percutaneous permeability of galanthamine cream.METHODS:Using iso?lated mouse's skin as barrier to permeation,the promoting effect of azone in different concentrations on permeation of galanth_ amine was studied.RESULTS:Azone could obviously enhence the permeability of galanthamine through skin.The steady flux of galanthamine cream containing2%and5%azone increased56.12%and23.29%respectively.CONCLUSION:Galanthamine cream has good percutaneous permeability and2%azone promotes the permeability best.

Objective To study the preparation and the in vitro and in vivo release profile of GH-Chitosan-Alginate microcapsules.Methods GH-Chitosan-Alginate microcapsules were prepared through impulsive electrostatic technique.The interrelated factors influencing the diameter and sphericity were studied through orthogonal experiments,and finally the statistic analysis made sure the optimum conditions to prepare microspheres.The morphology and size of the microcapsules were observed,and the content,encapsulation efficiency and recovery efficiency of the microcapsules were measured.Moreover,their in vitro and in vivo release experiments were carried out.Results The results showed that the diameter of needle was the most significant factor to the diameter of microspheres.The optimum conditions for the least diameter of microspheres were 450?m diameter of needle,2cm from needle tips to the gelation surfaee,1.5% alginate concentration,8ml/h speed of flowing-liquid and metal containers.The microcapsules had good sphericity morphology and distribution.The size of the microcapsules was in the range of 10-25?m with an average size of 47.93?m.The encapsulation efficiency and GH-load of the microcapsules were 94% and 11.24% respectively.The release kinetics of microcapsules was studied in false gastric and intestines juice.In false gastric juice,the GH of microcapsules was not released;in false intestines juice,it was released well,and TAM was completely released after about 12h.in vivo release profile made sure that the serum GH level of GH microcapsule group was at the highest value(98.59ng/ml) at 8h.The release profile was fitted well in both in vitro and in vivo conditions.Conclusion GH-Chitosan-Alginate Microcapsules have good morphology and sustained release effect.

OBJECTIVE: To study in vitro percutaneous permeability of methylphenidate cream. METHODS: Isolated rat skin was taken as permeable barrier. The influence of different concentrations of azone(0%, 2%. 5% ) in methylphenidate cream on drug permeation was observed. RESULTS: Steady-state percutaneous flow(J ) of methylphenidate cream with 2% and 5% azone increased 27. 80% and 49. 05%. respectively. CONCLUSION: Methylphenidate cream will be a safe. effective and conve- nient new preparation.

Objective To explore the effects of H.pylori and crude extracted proteins secreted by H.pylori(broth culture filtrate protein,BCF-P)on acid secretion from isolated rabbit parietal cells.Methods Parietal cells from rabbit gastric mucosa were isolated and enriched with digestion and elutriation.H.pylori(NCTC 11637,CagA+ VacA+)were grown in liquid broth culture and BCF-P was precipitated with ammonium sulfate.The vacuolation activity of BCF-P was evaluated with neutral red dye uptake test in HeLa cell.Isolated parietal cells were incubated with H.pylori(bacteria/cell=100∶1)for 2 h and 16 h,or BCF-P(100μg/ml)for 1 h and 12 h.Acid secretion from parietal cells was studied using 14C-aminopyrine(14C-AP)accumulation indirectly and H+-K+ ATPase α subunit mRNA expression was assessed using RT-PCR.Results (1)BCF-P containing vacuolating cytotoxin(VacA)with vacuolation activity on HeLa cells had positive result on neutral red uptake test.(2)The basal expression of H+-K+ ATPase α subunit mRNA could be detected in isolated parietal cells and 14C-AP accumulation was significantly increased in response to the stimulation of histamine with different concentrations for 30 min(P＜0.05).These results indicated that the isolated parietal cells retain relative intact acid secretion function.(3)The histamine(1.0×104 mol/L)stimulated acid secretion was inhibited sustainedly in response to H.pylori by 81% at 2 h and by 94% at 16 h(P＜0.05).However,H+-K+ ATPase α subunit mRNA expression was up-regulated in tlle acute period(2 h)and was down-regulated in the chronic period (16 h)by H.pylori(P＜0.05).(4)BCF-P significantly inhibited the histamine-stimulated acid secretion by 24% at 1 h and by 58% at 12 h(P＜0.05),and this inhibition was accompanied by the down-regulated expression of H+-K+ATPase α subunit mRNA.Conclusions Intact H.pylori and VacA secreted by H.pylori could directly inhibit histamine-stimulated acid secretion from parietal cells and this inhibition may be mediated by the down-regulated H+-K+ ATPase expression.

Anti-angiogenesis has been one of the important strategies in treating tumor metastasis,diabetic retinopathy,rheumatic arthritis etc.Angiogenesis models have played an active role in studying the mechanism of angiogenesis and developing drugs of pro-or anti-angiogenesis.A common problem faced by angiogenesis researchers has been the difficulty of choosing suitable models.This paper reviews principal models in use for some details and their advantages and shortages.

A series of valproic acid salicylanilide esters were designed and synthesized based on the principle of prodrug. The structures of the target compounds were confirmed by MS, 1H NMR and 13C NMR. Anti-tumor activities of these compounds against K562, A549, A431 cells in vitro were investigated by MTT assay and SRB assay. The results indicated that the compounds 6h-6j were found to have stronger cell growth inhibitory action than gefitinib, and comparable to niclosamide, which are worth to be intensively studied further.