In vivo anti-obesity effect of the agonist for receptor VPAC1 / 生理学报

It was hypothesized that the VPAC1 agonist may exert anti-obesity functions because VPAC1 is involved in the anorexigenic effects and the anti-inflammatory function of pituitary adenylate cyclase-activating polypeptide (PACAP)/vasoactive intestinal polypeptide (VIP). Furthermore, our in vitro test showed that the expression of VPAC1 increased significantly after the 3T3-L1 adipocytes were differentiated, and that incubation of adipocytes with VPAC1 agonist (10-1 000 nmol/L per 1x10(6) cells) resulted in stimulation of lipolysis. To test the effect of VPAC1 agonist [Lys15, Arg16, Leu27]-VIP (1-7) GRF (8-27) on diet-induced obesity (DIO), we further designed the following two in vivo experiments: (1) Mice were fed on high-fat diet (HFD) and intraperitoneally (i.p.) treated with VPAC1 agonist simultaneously for 28 d; (2) Mice were given HFD for 35 d, and subsequently fed on the same HFD and i.p. treated with VPAC1 agonist for the next 28 d. The physiological indices, including body weight, weight of white adipose tissue, plasma glucose and blood lipid, were collected. The results showed that treatment with VPAC1 agonist inhibited ingestion significantly and prevented the elevations in body weight and the weights of the white adipose tissues (epididymal and dorsal) induced by HFD. The increases in plasma glucose, cholesterol, triglycerides and LDL induced by HFD were also down-regulated in mice treated with VPAC1 agonist. VPAC1 agonist treatment also improved the glucose tolerance. Therefore, VPAC1 agonist treatment inhibits the development of the obesity induced by HFD and helps to improve the morbidities associated with DIO.

Expression and characterization of VPAC2 in CHO cells / 生物工程学报

VPAC2 is a co-receptor of pituitary adenylate cyclase activating polypeptide (PACAP) and vasoactive intestinal peptide (VIP) and mediates multiple bio-functions. In order to construct the CHO line expressing VPAC2 stably, pcDNA-VPAC2 was used to transfect CHO cells. The positive clones were selected by G418 and the clone VPAC2-CHO with high sensitivity to PACAP38 was picked out by its ability to promoting the concentration of cAMP. RT-PCR, Western blot and Immunofluorescenece assay were used to identify the express of VPACS. Binding competition with VPAC2 agonist and the bioactivity of mediating the ligand to promote the concentration of cAMP showed that VPAC2 was expressed effectively in VPAC2-CHO. The results of Scatchard analysis revealed that VAPC2-CHO expressed a receptor density of (1.1 +/- 0.2) pmol/mg protein, respectively, with Kd values of (0.55 +/- 0.10) nmol/L for PACAP38 used as a tracer. The construction of CHO cells expressing VPAC2 specially and functionally lays a foundation not only for the further research on the characters and functions of VPAC2 but also for the screening and characterization of novel agonists of antagonists for VPAC2.

Cloning and Expression of the New Gene Recombinant RMBAY Against Type-2 Diabetes and Its Production Optimization / 中国生物工程杂志

Constructing prokaryotic expression vector pKY-RMBAY by gene recombination and research its optimizing productive conditions.By PCR technology synthesizing the gene of the RMBAY with preference codon of E.coli and the RMBAY gene was inserted into high efficiency expression vector pKYB-MCS.Expressed fusion proteins in E.coli ER2566 were purified with Chitin-Beads column.Fusion proteins binding on Chitin-Beads was cut on N-terminus of intein due to the induction of ?-mercaptoethanol and the target peptide RMBAY was released.The RMBAY was identified by mass spectrum.Experiment results showed RMBAY can be high efficiently expressed in E.coli ER2566,with optimizing productive conditions the yield of the RMBAY may be 6.7mg/L fermentation product and its purity is greater than 98%.The molecular weight of RMBAY is 3.887 kDa by mass spectrum and that accords with its theory value.

Preparation and Identification of Recombinant Vasoactive Intestinal Peptide / 中国生物工程杂志

In order to prepare the recombinant vasoactive intestinal peptide (VIP) using intein mediated rapid purification system,the cDNA encoding the recombinant VIP was designed and synthesized according to the preference of E.coli,and then was cloned into the expression vector PTWIN. The recombinant plasmid PTWIN-VIP was transformed into expression host E.coli strain ER2566.The fusion protein consisting of the recombinant VIP,intein and chitin binding domain was expressed and purified by chitin affinity chromatography. The target peptide was released from the fusion protein by changing the temperature and the pH of the cleavage buffer. The molecular weight of the recombinant VIP was determined by the mass spectrometry and the results was conformity with the theoretical value. The preliminary bioactivity assay indicated that the recombinant VIP decreased the serum resistin levels significantly in LPS-induced acute inflammation. The preparation and the characterization of anti-inflammatory effects of the recombinant VIP layed the foundation for its further application.

Effects of the second renal transplantation on the sexual function of kidney recipients / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the effects of the second renal transplantation on sexual function.</p><p><b>METHODS</b>Thirty kidney graft recipients, including 29 cases of the second renal transplantation and 1 case of simultaneous dual kidney transplantation, responded to the questionnaire. The penis cavernosal artery flow of these patients were examined by color doppler ultrasonography. Of the 30 recipients, 9 underwent bilateral kidney transplantation with their bilateral external iliac arteries anastomosed to the donors' renal arteries (Group A), 10 recipients with their unilateral external iliac arteries and the other internal iliac arteries anastomosed to the donors' renal arteries (Group B), the other 10 with their internal iliac arteries anastomosed to the donors' renal arteries (Group C).</p><p><b>RESULTS</b>Eight recipients of Group A, 7 of Group B, and 5 of Group C were restored to normal sexual function 6 months after kidney transplantation. The peak systole velocity (PSV) in Group C was slower than in Groups A and B.</p><p><b>CONCLUSION</b>Kidney transplantation with the second internal iliac arteries anastomosed to donors' renal arteries may affect the sexual function of the recipients, but some might enjoy satisfactory sexual life some time after the establishment of lateral branch circulation.</p>

The effect of intermittent deferoxamine therapy on relieving iron overload in children with ? thalassemia / 实用儿科临床杂志

Objective To evaluate the effect of the intermittent deferomamine(DF) therapy on relieving iron overload caused by transfusion in children with ? thalassemia.Methods Sixteen children who were finally diagnosed as ? thalassemia major were treated with deferomamine for 124 times totally to low the iron overload. The serum iron(SI), serum ferritin(SF) and urine ferritin were detected each time with radio-immunity technique and difference was compared before and after treatment. Meanwhile, weather DF involved children′s liver and renal function was observed in whole procedure.Results Iron overload exists in 16 cases of ? thalassemia major children by a long- term hypertransfusion therapy, with average level SI 33.69?6.72 mmol/L,SF 441.19? 54.70 ?g/L,urine ferritin 8.64?6.79 ?g/L. The difference was significant (paired-samples t test,t =6.173 P 0.05).Conclusion The study suggest that intermittent low-dose DF therapy is effective for iron overload caused by transfusion in ? thalassemia children, without apparent side effects.