GLP-1 has a variety of anti-diabetic effects. However, native GLP-1 is not suitable for treatment of diabetes due to its short half-life (t½, 2-5 min). Exendin-4 is a polypeptide isolated from lizard saliva, which can bind to GLP-1 receptor, produce physiological effects similar to GLP-1, t½ up to 2.5 h, therefore, we developed a long-lasting GLP-1 receptor agonists and GLP-1-exendin-4 fusion IgG4 Fc [GLP-1-exendin-4/ IgG4(Fc)]. We constructed the eukaryotic expression vector of human GLP-1-exendin-4/IgG4(Fc)-pOptiVEC- TOPO by gene recombination technique and expressed the fusion protein human GLP-1-IgG4 (Fc) in CHO/DG44 cells. The fusion protein stimulated the INS-1 cells secretion of insulin, GLP-1, exendin-4 and fusion protein in CD1 mice pharmacokinetic experiments, as well as GLP-1, exendin-4 and fusion protein did anti-diabetic effect on streptozotocin induced mice. Results demonstrated that the GLP-1-exendin-4/IgG4(Fc) positive CHO/DG44 clones were chosen and the media from these positive clones. Western blotting showed that one protein band was found to match well with the predicted relative molecular mass of human GLP-1-exendin-4/IgG4(Fc). Insulin RIA showed that GLP-1-exendin-4/IgG4(Fc) dose-dependently stimulated insulin secretion from INS-1 cells. Pharmacokinetic studies in CD1 mice showed that with intraperitoneal injection (ip), the fusion protein peaked at 30 min in circulation and maintained a plateau for 200 h. Natural biological half-life of exendin-4 was (1.39 ± 0.28) h, GLP-1 in vivo t½ 4 min, indicating that fusion protein has long-lasting effects on the modulation of glucose homeostasis. GLP-1-exendin-4/IgG4(Fc) was found to be effective in reducing the incidence of diabetes in multiple-low-dose streptozotocin-induced diabetes in mice, longer duration of the biological activity of the fusion protein. The biological activity was significantly higher than that of GLP-1 and exendin-4. GLP-1-exendin-4/IgG4(Fc) has good anti-diabetic activity. It can be used as a long-acting GLP-1 agonists.
Animals ; CHO Cells ; Cricetinae ; Cricetulus ; Diabetes Mellitus, Experimental ; drug therapy ; Glucagon-Like Peptide 1 ; pharmacology ; Glucagon-Like Peptide-1 Receptor ; agonists ; Half-Life ; Humans ; Hypoglycemic Agents ; pharmacology ; Immunoglobulin G ; pharmacology ; Insulin ; secretion ; Mice ; Peptides ; pharmacology ; Recombinant Fusion Proteins ; pharmacology ; Venoms ; pharmacology

Objective To study the imaging of inner ear malformations in children with congenital sensorineural hearing loss(SNHL).Methods CT and MRI examinations were performed on children with SNHL diagnosed by audiological test.One hundred and eighty-eight patients with complete imaging information were obtained.The imaging of inner ear malformations was analyzed according to Sennaroglu's classification. Results Thirty-five patients(54 ears) were found with inner ear malformations by CT and MRI,3 of whom(5 ears) were accompanied by outer and middle ear malformations.Among the 35 patients,2(4 ears) were found to be common cavity deformity,1(1 ear) cochlear hypoplasia,13(26 ears) incomplete partition II(Mondini deformity),4(7 ears) vestibule dilation,13(19 ears) semicircular canal deformity and 19(34 ears) large vestibular aqueduct.Internal auditory canal was found narrow in 7 patients(10 ears) and wide in 1(2 ears) with classic Mondini deformity. Conclusion CT and MRI examinations are of great importance to the diagnosis and treatment of inner ear malformations in children with congenital SNHL,especially for the candidates of cochlear implantation.

Objective To develop a rapid, sensitive and specific assay based on multiplex real-time PCR for detecting and identifying Escherichia coli O157: H7. Methods The lipopolysaccharide gene (rJbE) and H7 flagellar antigen gene(fliC) of Escherichia coli O157:H7 was chosen as targets, and then the primers and TaqMan-MGB probe were designed. The 5'end of probes was labeled with FAM and HEX fluo-resceins respectively; the 3'end of probes was labeled with MGB. The PCR reaction was optimized systemati-cally. Then the specificity, sensitivity and reproducibility of multiplex real-time PCR were estimated. Final-ly, multiplex real-time PCR was applied to detected clinical specimens. Results Escherichia coil O157:H7 were detected by multiplex real-time PCR accurately and quickly, which could distinguish Escherichia coli O157:H7 from O157: non-H7. Meanwhile, none of other bacteria could be identified. The sensitivity was 10 CFU/ml in pure culture. The coefficient of variation of intra-assay and inter-assay was less than 5%. When this assay was applied directly to identify 66 clinical specimens, the results showed that t5 were positive to Escherichia coil O157:H7 and 2 were positive to Escherichia coil O157: non-H7, in which 16 was the same to the results obtained from the conventional assays. The coincidence was 98.49%. Conclusion It is showed that multiplex real-time PCR is a reliable, accurate and feasible assay for detecting and identifying Escherich-ia coli Oi57: H7, The assay reported here provided a tool for analysis and diagnosis in the field of detecting clinical pathogens, epidemiologic survey and food safety monitoring.

To compare the curative effects of two surgical treatment methods for open angle glaucoma and cataract patients.
●METHODS: Totally 69 patients with primary open angle glaucoma and cataract were divided into two groups:group A and group B. Thirty- four patients (34 eyes) in group A underwent small incision non -phacoemulsification cataract surgery combined with trabeculectomy, just combined operations, 35 patients (35 eyes) in group B underwent small incision non -phacoemulsification cataract surgery beyond 6mo after trabeculectomy, just two stage operations. Postoperative intraocular pressure at 1wk and 3mo, postoperative corrected visual acuity and filtering bleb at 3mo were observed, and intraoperative and postoperative complications were compared. All the patients were followed up for 3-6mo.
● RESULTS: All the visual acuity were increased compared with that before surgery in two groups, the difference was not statistically significant. All the intraocular pressure was controlled in two groups, the difference was not statistically significant, and the difference about the filtering bleb was neither statistically significant ( P > 0. 05 ). There were no serious intraoperative and postoperative complications in two groups.
● CONCLUSlON: The two stage operations and the combined operations both can control theintraocular pressure and improved visual acuity of open angle glaucoma and cataract. The two stage operations may prevent a second operation for post trabeculectomy cataract, allowing earlier visual rehabilitation.

Objective To explore the clinico-biological characteristics, treatment and prognosis of salivary duct carcinoma. Methods This study included 12 cases of salivary duct carcinoma treated in our hospital. Clinical data were retrospectively analyzed for patients admitted between April 1995 and October 2006. The clinical characteristics, histological features, imaging, therapy methods and prognosis were analyzed. Results Of 12 salivary duct carcinoma, there were 10 males, 2 females. The age of onset ranged from 53 to 73 year old and the average was 56 year old. Physical examination revealed a firm and unboundary mass accompanied by nerve infiltrating symptom. The histological appearance was characterized by solid cell nests with ductal structures and central comedonecrosis. Extensive resection and radical neck dissection was performed in 11 patients, postoperation radiation done in 10 patients and chemotherapy in 3 patients. One year survival rate was 83. 33% , that of 3 years was 41. 67% , and of 5 years was 25. 00% , the median survival time was 36 months. Conclusion Salivary duct carcinoma is a rare malignant salivary tumor and most patients are men. Regional extensive resection and postoperative radiation or chemotherapy are the mainstay of therapy. Lymph node metastases in level Ⅰ ,Ⅱ , Ⅲ are a common finding in patients with SDC and the prognosis is poor.

OBJECTIVETo explore the clinical symptom and effect of arthroscopic treatment of symptomatic anterior cruciate ligament (ACL) cysts of the knee.

METHODSClinical data from 12 symptomatic ACL cysts patients from January 2005 to December 2010 were retrospectively analyzed,including 8 males and 4 females,with an average age of (33.7±9.5) years old (ranged, 19 to 53 years old). The locations were the left knee in 5 cases and the right knee in 7 cases. The disease duration ranged from 3 to 48 months,with a mean of (15.8±13.2) months. All cysts were arthroscopically resected. Range of motion was measured preoperatively and postoperatively, and Lysholm scoring system was used to evaluate the knee function.

RESULTSAll the incisions healed by first intention, and no complications occurred. Twelve patients were followed up for an average of (32.3±6.6) months(ranged, 24 to 48 months). The symptoms of arthralgia,swelling and interlocking of the affected knees disappeared. There was no recurrence during the follow-up. There were significant differences in the range of motion and Lysholm score between pre-operation and post-operation.

CONCLUSIONArthroscopic surgery, showing its advantages of minimal invasion and rapid recovery,is an effective measure in the treatment of ACL cysts.

Adult ; Anterior Cruciate Ligament ; physiopathology ; surgery ; Arthroscopy ; methods ; Cysts ; physiopathology ; surgery ; Female ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Range of Motion, Articular

Objective To summarize the susceptible factors and the experience of the surgical treatment of ruptured aneurysm of the sinus of Valsalva combined with infective endecarditis.Methods From January 2000 to March 2008,30 cases with raptured aneurysm of the sinus of Valsalva were admitted, clinical data of 10 cases combined with infective endocarditis were retrospectively reviewed.Nine cases underwent the surgical treatment,including 6 cases accepted aortic valve replacement (AVR).Results Nine eases underwent the surgical treatment followed up for 2 months to 8 years,cardiac function of the survivors recovered to Class Ⅰ-Ⅱ in NYHA,7 cases were Ⅰ grade,2 cases were Ⅱ grade.One case was dead for MSOF not underwent the surgical treatment.Conclusions Associated with ventricular septal defect is one of the susceptible factors.Following diagnosis of ruptured aneurysm of sinus of Valsalva combined with infective endocarditis,sufficiently and validly antibiotic and surgical treatment should be pedormed as quickly as possible,and AVR to the moderate and severe aortic insufficiency.

OBJECTIVE: To investigate the regulation of hepatitis C virus (HCV) core protein on the activity of signal transducers and activators of transcription 3 (stat3). METHODS: A cell line expressing stable HCV core protein-QSG7701-core was constructed by transfecting the pcDNA3. 1-core (expressing HCV core protein) into the human immortalized hepatocyte line QSG7701. The phosphorylation and DNA binding activity of stat3 were detected by immunocytochemistry, Western blot, and electrophoretic mobility shift assay (EMSA). RESULTS: The expression level of phosphorylated stat3 in QSG7701-core cells was significantly lower than that in QSG7701-pcDNA3. 1 cells and untransfected QSG7701 cells, but there were no significant differences in the expression levels of total stat3 among the 3 groups. The positive signal of phosphorylated stat3 in nucleus of QSG7701-core cells was obviously weaker than that in QSG7701-pcDNA3. 1 cells and untransfected QSG7701 cells. EMSA showed that DNA binding activity of stat3 in QSG7701-core cells significantly decreased. CONCLUSION The expressionof HCV core protein in human hepatocyte line may suppress the phosphorylation and DNA binding activity of stat3, which may be one of the causes for resistance against interferon.
Cell Line ; DNA-Binding Proteins ; metabolism ; Hepatocytes ; cytology ; metabolism ; Humans ; STAT3 Transcription Factor ; metabolism ; Signal Transduction ; Transfection ; Viral Core Proteins ; biosynthesis ; genetics

OBJECTIVETo compare the clinical efficacy between total aortic arch reconstruction with open triple-branched stent graft placement and total aortic replacement combined with stented elephant trunk implantation for patients with Stanford A aortic dissection.

METHODSPatients with Stanford A aortic dissection treated with surgical treatment from January 2006 to May 2011 were included in this study. The patients were divided into two groups. Group I (n = 20) patients were treated by total arch replacement with stented elephant trunk procedure. Group II (n = 8) patients received open triple-branched stent graft placement. Echocardiography and aortic CT angiography were performed before and at 1 month after operation.

RESULTSAge, gender and disease severity were similar between the 2 groups (all P > 0.05). Operation was successful in all 28 patients. Cardiopulmonary bypass time, aortic cross clamp time, circulation arrest time and duration of ventilator assisted breathing were significantly longer; postoperative drainage volume and blood transfusion volume were significantly larger and hospitalization cost was significantly higher in group I patients compared those in group II patients (all P < 0.05). One month after operation, the maximum internal diameter of aorta was smaller than pre-operation in both group I [(30.2 ± 3.1) mm vs. (42.5 ± 6.5) mm, P < 0.05] and group II [(31.5 ± 2.5) mm vs. (44.1 ± 7.3) mm, P < 0.05].

CONCLUSIONSShort-term procedural success rate was similar between the two groups. The total aortic arch reconstruction with open triple-branched stent graft placement procedure is simpler, shortens the operation time, reduces the blood transfusion volume and is more cost-effective compared to the classical aortic arch operation.

Adult ; Aneurysm, Dissecting ; surgery ; Aorta, Thoracic ; surgery ; Aortic Aneurysm ; surgery ; Female ; Humans ; Male ; Middle Aged ; Stents ; Treatment Outcome

OBJECTIVETo determine lobetyolin in the root of Codonopsis tangshen from the various cultivation areas.

METHODA Supelco Discovery C18 Column (4.6 mm x 250 mm, 5 microm) was used with acetonitrile-0.5% acetic acid in water (20:80) as the mobile phase and UV detection was at 268 nm.

RESULTTwenty-four batches of the samples were analyzed. The content of lobetyolin ranged from 0.0403-0.9667 mg x g(-1).

CONCLUSIONThe method was simple, reproducible and reliable. It can be used to control the quality of C. tangshen.

China ; Chromatography, High Pressure Liquid ; methods ; Codonopsis ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Plant Roots ; chemistry ; Polyacetylenes ; analysis ; Quality Control