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.

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.

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 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.

Objective To investigate the therapeutic effect of immunodepressant on Crohn's disease (CD).Methods 105 patients with CD were collected from 1983 to 2006 in Peking Union Medical College Hospital.All of their clinical manifestations and therapeutic results were analyzed retrospectively.Results (1)The application of immunedepressant was significantly increased after the year of 2000 (34.7% vs 3.0%,P=0.000).(2)The application of immunodepressant and the clinical features of these patients were as fouows：①The number of patients with modcrate to severe CD were more than that with mild CD in those using immunodepressant (28.9% vs 0).②The use of immunodepressant was not related with the diseased site of CD.because there were no difference among the groups with lesion in small intestine (20.0%),colon(27.3%)and ileocolon(27.1%),P=0.726.③The serum albumin level of CD patients using immunodepressant was significantly lower than that of those not using(31.9 g/L vs 35.1 g/L,P=0.047).④The use of immunodepressant did not decrease the incidenee of operative treatment(38.5% vs 50.0%,P=0.320).(3)The rate of remission in 19 CD patients using azathioprine is 68.4%(13/19)and the percentage of neutrophil in the group with relief was lower than that without relief(0.76 vs 0.65,P=0.032).Conclusions Immunodepressant is playing more important role in the treatment of CD.The patients with moderate and severe CD with as well as lower serum albumin should be treated with immunodepressant as early as possible.Whether immunedepressant is necessary or not is not decided by the diseased site.CD patients with higher percentage of neutrophil have less therapeutic effect than those with lower.

Objective To investigate the causes, clinical features, treatment and prognosis of gastrointestinal bleeding (GIB) patients in emergency department.Methods To analyze prospectively the clinical data of 168 GIB patients admitted to the emergency department of Peking Union Medical College Hospital during 2006.1-2006.12.Results (1) General data; male: female = 1.75:1 ( 107: 61) , mean age 13-87(56.5 ±17.8) years with a peak in 60-69 years.The percentage of old patients was significantly higher than that of young and middle age ( 52.4% vs 19.6% and 28.0% , P = 0.000 ).( 2 ) The incidence of acute gastric mucosal lesion in patients taking non-steroidal antiinflammatory drugs ( NSAIDs) ( 18.5% ) was significantly higher than that in patients not taking( 0.7% , P = 0.000 ).( 3 ) 86.9% ( 146/168 ) of the patients had anemia.(4) More patients who took emergency gastroscopy could be diagnosed than those patients who did not (89.4% vs 58.5% , P =0.000), while no significant difference could be seen between patients who took emergency enteroscopy and patients who had non-emergency gastroscopy (20.0% vs 57.9% , P =0.315).(5)The hemostatic ratio in GIB patients due to peptic ulcer was obviously higher than that in GIB patients due to other causes (86.0% vs 40.7% ,P =0.000).The rate of emergency operation for GIB patients was 1.8%.Conclusions Most of the GIB patients admitted to tertiary general hospitals are elderly males.NSAIDs administration is one of the most important causes of upper GIB.Upper GIB patients should have gastroscopy as soon as possible, while emergency coloscopy is of little significance in cases with lower gastrointestinal hemorrhage.

Objective Matrix-assisted laser desorption ionization-time of might-mass spectrometry (MALDI-TOF-MS) was utilized to analyze the protein fingerprint in brain-gut interaction of irritable bowel syndrome (IBS) model rats' colon, so as to find the clues for IBS. Methods Fourteen healthy male adult Wistar rats were selected and divided into a control and a chronic and acute stress ( CAS) group. Colon motility, visceral sensation and behavior changes of rats were detected to evaluate the model. MALDI-TOF-MS was used to observe the overall view of protein in colon so as to study whether there are abnormalities of protein levels in IBS. Results As compared with those in the control group, the number of fecal pellets [ (6. 00 ± 1. 69 ) pellets/1 h vs ( 1. 14 ± 0. 69 ) pellets/1 h, P < 0. 01 ] and frequency of abdominal contraction induced by colorectal distention (CRD) increased, while the amount of weight gain [ (298. 88 ± 18.61)gvs (348. 00±12. 44)g, P<0.01] and consumption of sucrose solutions [ (13. 63 ± 1. 69) ml/1 h vs (19.00±3.06) ml/1 h, P<0.05] decreased in the CAS group (P <0. 05). As far as protein/peptide quality different peak was concerned, CAS rats had 12 different peaks compared with the control rats. The different proteins could be divided into 4 types, which were related to iron secretion, protein synthesis, G protein system and immunity. The protein levels of the model group were higher than those in the control group (P < 0. 05). Conclusions The CAS rats integrate the major characteristics of IBS such as altered colon motility, higher visceral hypersensitivity and psychiatric disorder and can mimic the brain-gut interaction of IBS partly. The detection of differential proteins provides reference for the pathogenesis and treatment of IBS.