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 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 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 investigate the possible influence of immunosuppressive therapy,including sirolimus (SRL) and calcineurin inhibitors (CNI, tacrolimus), on level of Treg in the liver allo-graft recipients, Methods Forty-seven liver transplant recipients with stable liver function were assessed for at least 2 years, and divided into two groups: one composed of 15 patients receiving SRL,and another, of those receiving CNI (32 patients with tacrolimus). Thirty-eight age-matched healthy subjects (HS) were used as normal controls. We examined the expression of CD4, CD25, and Foxp3 in peripheral blood monouclear cells. Treg of every group was analyzed using Cell Quest software.Results SRL significantly increased the prevalence of CD4+ CD25high T cells, compared with HS and CNI group. The prevalence of CD4+ CD25high T cells of CNIs group was significantly lower than that of HS. The percentage of CD4+ CD25high T cells in the total CD4+ T cells was 1.88 % (1.56 %-2. 60 %), 1. 15 % (0. 57 %-l. 48 %) and 0. 84 % (0. 46 %-1. 45 %) in SRL, NS and CNI group,respectively (P<0. 01 or P<0. 05). Foxp3 was expressed in more than 95 % of CD4 + CD25high T cells and less than 20 % of CD4+ CD25low T cells, whereas not expressed in CD4+ CD25- T cells.Conclusion Different immunosuppressive therapy (SRL or CNI) might have different roles in tolerance induction in liver transplant recipients. Namely, SRL promoted the induction of allo-antigen tolerance, but CNI hampered the progression.

Objective To investigate the clinical value of serum VEGF levels in the diagnosis and treatment of colon cancer.Methods Serum VEGF was detected by ELISA,and CEA and CA199 concentration were detected by CLEIA in 66 patients with colon cancer,55 patients with colon benign diseases and 50 health persons.The value of individual and joint detection for VEGF in colon cancer was evaluated.Analysis had been done on relationships between serum VEGF and pathology,treatment effects and prognosis.Results The levels of serum VEGF in colon cancer group [(318.5±148.6) ng/L] were significantly higher than those in control group [(125.7±49.4) ng/L] and benign colon diseases [(136.9±52.6) ng/L] (t =8.830,8.805,all P ＜ 0.01).There was a positive correlation between serum levels of VEGF and depth of tumor size,tumor invasion,lymph node metastasis and TNM stage (P ＜ 0.01).The susceptibilities of VEGF,CEA,CA199 were 61％,45 ％,53 ％.The sensitivity of detection was improved to 86 ％ when the combined detection of VEGF,CEA and CA199 (x2 =11.237,P ＜ 0.01).The serum levels of VEGF in patients with colon cancer was significantly decreased after treatment in the 3,7,10 day compared with that before operation [(272.3±88.1),(236.8±77.4),(173.1±59.9) vs (318.5±148.6) ng/L,t=2.173,P ＜ 0.05; t =3.961,P ＜ 0.01; t=7.464,P ＜ 0.01],respectively.Conclusion The VEGF was related to the onset and progression and metastasis of colon cancer.It has clinical significancy for diagnosis of colon cancer and judgment of curative effect and prognosis.

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.