The effect of continuous amylin infusion on glucose tolerance in rats and the effect of amylin on glucose-stimulated insulin secretion from isolated rat pancreatic islets were sudied. Three groups of animals formed the study of glucose tolerance: Group 1 received salined infusion; Group2 , amylin at 2. 6 nmol?kg-1/h;and group 3,amylin at 26 nmol**kg-1/h. The total infusion time was 60 min. The insulin level and serum glucose were not altered at lower dose of amylin. However glucose levels markedly rose at 2.7,12 min. after beginning IVGTT, without significant change of insulin levels in group 3. The response of insulin to 16. 7 mol glucose-stimulated isolated rat pancretic islets was singnificantly inhibiled by the addition of amylin 10 umol.

Sodium orthovanadate is one of vanadate salts. When administered orally to STZ-induced diabetic rats, sodium orthovanadate may stimulate glucose uptake and. metabolism and made them normoglycemic. However, the blood glucose levels of normal rats treated with sodium orthovanadate did not change despite their plasma insulin levels decreased. The insulin-like effect of sodium orthovanadate on carbohydrate metabclism was not due to direct stimulation of insulin secretion, because the plasma insulin concentration of vanadate-treated STZ-rats remained lower and insulin secretion of isolated pancreas perfused with solution containing sodium orthovanadate was smilar to that of controls.

OBJECTIVE: To discuss the way to evaluate outpatient prescription in medical institutions according to Prescription Administrative Policy so as to provide reference for the practice of prescription evaluation system. METHODS: 500 outpatient prescriptions from April 16 to April 20 in 2007 in our hospital were sampled randomly for an evaluation in respect of drug utilization, prescription behavior, pharmaceutical care level etc. RESULTS & CONCLUSIONS: To evalnation from The prescriptions involved use of injections and of antibacterials, prescribed drugs were covered in "national essential drugs" , prescriptions with general name, the knowledge of patients on the administration and dosage were drectively.It is supply reference for the prescription evaluation system.

Objective:To investigate outpatient's medicine knowledge and satisfaction degree and analyze their correlation.Method:150 outpatients,their doctors and pharmacists were investigated with a fieldwork method.Patients' medical knowledge and satisfaction degree were measured with a quantitative scale table,and their correlation was calculat- ed with a linear regression method.Result:Almost all the doctors and pharmacists could provide their patients with their guidance about medicines,which focused on drug dosage and taking ways.Fewer than 20% patients could take medical ad- vice of potential risks and notice of taking medicine.A significant correlation was shown between patient's medical knowl- edge and satisfaction degree(r=0.76).Conclusion:More guidance from doctors and pharmacists should be provided for outpatients.The medical knowledge learned by patients could influence patient's satisfaction degree significantly.

Objective To evaluate the value of miniprobe sonography (MPS) in diagnosing gastrointestinal submucosal protrusive lesions and selecting the indicated manual for treatment. Methods According to the sizes, properties, and depth of SMTs in the gastrointestinal tract detected by the MPS, different methods of resection were performed. Results Of 24 cases, 11 SMTs lying in submucosa under 2cm in diameter (2 benign gastrointestinal stromal tumors, 3 lipomas, 5 cysts, 1 granular cell tumor) were attempted with EMR or argon plasma coagulation (APC) ; there were no complications of hemorrhage or perforation. Thirteen SMTs lying in muscularis propria or with size of SMTs above 2 cm in diameter (4 malignant GISTs, 6 benign GISTs, 1 lipoma, 2 aberrant pancreas) were performed by surgical resection. Preoperative diagnoses of SMTs by MPS were consentient with their histological diagnoses. Conclusion MPS may detect the size, property, and depth of SMTs in the gastrointestinal tract and is helpful in selecting indicated cases for endoscopic resection. Endoscopic therapy of SMTs lying in mucosa or submucosa under 2cm in diameter is a safe and effective procedure.

Objective To construct a lentiviral vector carrying rat sirt1 gene and observe the expression of sirt1 in retinal ganglion cell (RGC) of rat.Methods Rat sirt1 cDNA was inserted into pLV5 vector.After identification by sequencing analysis and PCR,the recombinant sirt1expressinglentivirus vector was packaged by cotransfecting 293T cells with packaged plasmid.Then pLV5-sirt1 was used to infect the cultured Sprague-Dawley rat RGC cell in vitro.The expressions of sirt1 protein and mRNA in infected rat RGC were detected by quantitative real-time PCR and Western blot.Results The sirt1 expression vector pLV5 was successful constructed and sequence was proved to be correct.The expression of sirt1 protein and mRNA in RGC was significantly increased than that in cells infected with control lentiviruses (P ＜ 0.05).Conclusion We have successful constructed a sirt1 expression lentivirus vector pLV5-sirt1 and it can increase the expression of sirt1 protein and mRNA in the rat retinal ganglion cells.

OBJECTIVETo establish a canine cell line with p53 gene knockdown by lentivirus- mediated RNA interference (RNAi).

METHODSFour pairs of oligonucleotide sequences of p53 gene were synthesized and cloned into the pGMLV-SC5 RNAi vector. Following confirmation by PCR and DNA sequencing, the recombinant pGMLV-p53 plasmids and packaging mix vectors were packaged into mature lentivirus to infect 293T cells. The supernatant of the infected cells was harvested to infect WRD cells, in which p53 expression was detected using Western blotting and real-time PCR. The lentivirus with the strongest p53⁻ silencing effect was packaged for infecting WRD cells at the optimal multiplicity of infection (MOI) to establish a stably infected cell line (WRD/p53⁻) screened using puromycin.

RESULTSThe lentivirus carrying p53 shRNA was constructed successfully and a virus titer of 1 × 10⁹ TU/ml. The 4 pGMLV-p53 plasmids all produced strong p53 interference affects, and pGMLV- p53A1 with the strongest effect. The stably infected cells line WRD/p53⁻ was established successfully using pGMLV-p53A1 plasmid.

CONCLUSIONThe canine cell line WRD/p53⁻ with stable lentivirus-mediated p53 silencing has been established successfully.

Animals ; Cell Line ; Dogs ; Gene Knockdown Techniques ; Genes, p53 ; Genetic Vectors ; Lentivirus ; Plasmids ; RNA Interference ; RNA, Small Interfering ; Real-Time Polymerase Chain Reaction

Objective To establish a canine cell line with p53 gene knockdown by lentivirus-mediated RNA interference (RNAi). Methods Four pairs of oligonucleotide sequences of p53 gene were synthesized and cloned into the pGMLV-SC5 RNAi vector. Following confirmation by PCR and DNA sequencing, the recombinant pGMLV-p53 plasmids and packaging mix vectors were packaged into mature lentivirus to infect 293T cells. The supernatant of the infected cells was harvested to infect WRD cells, in which p53 expression was detected using Western blotting and real-time PCR. The lentivirus with the strongest p53-silencing effect was packaged for infecting WRD cells at the optimal multiplicity of infection (MOI) to establish a stably infected cell line (WRD/p53-) screened using puromycin. Results The lentivirus carrying p53 shRNA was constructed successfully and a virus titer of 1 × 109 TU/ml. The 4 pGMLV-p53 plasmids all produced strong p53 interference affects, and pGMLV- p53A1 with the strongest effect. The stably infected cells line WRD/p53- was established successfully using pGMLV- p53A1 plasmid. Conclusion The canine cell line WRD/p53-with stable lentivirus-mediated p53 silencing has been established successfully.

Objective To establish a canine cell line with p53 gene knockdown by lentivirus-mediated RNA interference (RNAi). Methods Four pairs of oligonucleotide sequences of p53 gene were synthesized and cloned into the pGMLV-SC5 RNAi vector. Following confirmation by PCR and DNA sequencing, the recombinant pGMLV-p53 plasmids and packaging mix vectors were packaged into mature lentivirus to infect 293T cells. The supernatant of the infected cells was harvested to infect WRD cells, in which p53 expression was detected using Western blotting and real-time PCR. The lentivirus with the strongest p53-silencing effect was packaged for infecting WRD cells at the optimal multiplicity of infection (MOI) to establish a stably infected cell line (WRD/p53-) screened using puromycin. Results The lentivirus carrying p53 shRNA was constructed successfully and a virus titer of 1 × 109 TU/ml. The 4 pGMLV-p53 plasmids all produced strong p53 interference affects, and pGMLV- p53A1 with the strongest effect. The stably infected cells line WRD/p53- was established successfully using pGMLV- p53A1 plasmid. Conclusion The canine cell line WRD/p53-with stable lentivirus-mediated p53 silencing has been established successfully.